FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Falloon, J Gallicano, KD Burstein, AH Piscitelli, SC AF Falloon, J Gallicano, KD Burstein, AH Piscitelli, SC TI Garlic supplements and saquinavir - Reply SO CLINICAL INFECTIOUS DISEASES LA English DT Letter ID HIV-INFECTED PATIENTS C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. Pfizer Gloval Res & Dev, Groton, CT USA. Tibotec Virco US, Durham, NC USA. Axelson Biopharma Res, Vancouver, BC, Canada. RP Falloon, J (reprint author), NIAID, Immunoregulat Lab, NIH, Bldg 10,Rm 11C103,10 Ctr Dr,MSC 1880, Bethesda, MD 20892 USA. NR 5 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD AUG 1 PY 2002 VL 35 IS 3 BP 343 EP 343 DI 10.1086/340988 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 573QA UT WOS:000176841300022 ER PT J AU Ziemann, U Tam, A Butefisch, C Cohen, LG AF Ziemann, U Tam, A Butefisch, C Cohen, LG TI Dual modulating effects of amphetamine on neuronal excitability and stimulation-induced plasticity in human motor cortex SO CLINICAL NEUROPHYSIOLOGY LA English DT Article DE excitability; stimulation-induced plasticity; amphetamine; human motor cortex; transcranial magnetic stimulation; ischemic nerve block ID LONG-TERM POTENTIATION; TRANSCRANIAL MAGNETIC STIMULATION; ISCHEMIC NERVE BLOCK; VISUAL-CORTEX; INTRACORTICAL INHIBITION; SYNAPTIC PLASTICITY; SENSORIMOTOR CORTEX; RAPID MODULATION; PHYSICAL THERAPY; CEREBRAL-CORTEX AB Objectives: The objective of the present study is to test the modulating effects of dextro-amphetamine (d-AMP) on excitability and stimulation-induced plasticity in human motor cortex. Methods: Transcranial magnetic stimulation (TMS) was used to treasure motor threshold, motor evoked potential (MEP) size and paired-pulse intracortical facilitation (ICF) in the biceps muscle of 7 healthy subjects before and after two different experimental manipulations: temporary forearm ischemic nerve block (INB) alone, or INB plus 0.1 Hz repetitive TMS (INB + rTMS) of the motor cortex contralateral to INB. Both manipulations were run after treatment with 10 mg of d-AMP or placebo (PBO). Results: In the PBO experiments, INB alone had no significant effect on MEP size or ICF, while INB + rTMS produced long-lasting (>60 min) increases. Compared with PBO, d-AMP led to a short-lasting (similar to10 min) increase in MEP size in the INB alone experiment, but suppressed the long-lasting increases of MEP size and ICF in the INB + rTMS experiment. Conclusions: The present findings suggest that d-AMP increases neuronal excitability but suppresses long-lasting stimulation-induced plasticity in human motor cortex. These dual effects may be relevant when using d-AMP to modulate human cortex function, (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Frankfurt, Neurol Clin, D-60590 Frankfurt, Germany. NINDS, Human Cortical Physiol Sect, NIH, Bethesda, MD 20892 USA. RP Ziemann, U (reprint author), Univ Frankfurt, Neurol Clin, Theodor Stern Kai 7, D-60590 Frankfurt, Germany. NR 48 TC 27 Z9 27 U1 0 U2 2 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 1388-2457 J9 CLIN NEUROPHYSIOL JI Clin. Neurophysiol. PD AUG PY 2002 VL 113 IS 8 BP 1308 EP 1315 AR PII S1388-2457(02)00171-2 DI 10.1016/S1388-2457(02)00171-2 PG 8 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 584YN UT WOS:000177496500017 PM 12140012 ER PT J AU Gordon, SM Brahim, JS Rowan, J Kent, A Dionne, RA AF Gordon, SM Brahim, JS Rowan, J Kent, A Dionne, RA TI Peripheral prostanoid levels and nonsteroidal anti-inflammatory drug analgesia: Replicate clinical trials in a tissue injury model SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Article ID ACUTE POSTOPERATIVE PAIN; ANTIINFLAMMATORY DRUGS; KETOROLAC TROMETHAMINE; PROSTAGLANDIN E-2; IMMUNOREACTIVE BRADYKININ; PHARMACOKINETICS; FLURBIPROFEN; METABOLISM; INHIBITION; IBUPROFEN AB Background: Nonsteroidal anti-inflammatory drug (NSAID) analgesia is generally attributed to peripheral suppression of cyclooxygenase (COX) enzymes, leading to decreased products of the arachidonic acid cascade. This study evaluated the in vivo relationship between levels of prostanoids at the site of tissue injury and analgesia after systemic or local NSAID administration in a clinical model of tissue injury. Methods: Subjects in two replicate clinical trials had one or two mandibular third molars removed and a microdialysis probe implanted at the surgical site for measurement of immunoreactive prostaglandin E, (PGE(2)) or immunoreactive thromboxane B-2 (TxB(2)) and pain measured concurrently. In the first study, ketorolac tromethamine (INN, ketorolac) was administered at pain onset in a 30-mg intramuscular dose, a 1-mg intramuscular dose, or a 1-mg submucosal dose at the extraction site in comparison with placebo. In the second study, subjects received either ketorolac tromethamine 30 mg by the intravenous route or placebo at pain onset. Results: PGE, was detectable in the first postoperative sample, decreased over the next hour, and then increased significantly coincident with the onset of postoperative pain. Administration of 30 mg ketorolac tromethamine produced parallel decreases in pain, PGE, levels, and TxB(2), levels at the surgical site. Administration of 1 mg ketorolac tromethamine intramuscularly or directly at the surgical site was analgesic but without measurable effects on PGE, levels. Conclusion: The temporal profile of PGE(2) and TxB(2) in the immediate postoperative period is consistent with constitutive COX-1 initially, followed by an increase in PGE2 resulting from expression of COX-2. The temporal association between NSAID analgesia and decreased prostanoids at the site of injury is consistent with a dual COX-1/COX-2 peripheral site of action. The analgesic effects of 1 mg ketorolac tromethamine without a reduction in PGE2 at the site of injury suggests an additional central site for NSAID analgesia. C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. RP Gordon, SM (reprint author), 30 Convent Dr,Bldg 30,Rm 106, Bethesda, MD 20892 USA. NR 32 TC 43 Z9 44 U1 0 U2 5 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD AUG PY 2002 VL 72 IS 2 BP 175 EP 183 DI 10.1067/mpc.2002.126501 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 592PM UT WOS:000177946100008 PM 12189364 ER PT J AU Lakatta, EG Sollott, SJ AF Lakatta, EG Sollott, SJ TI Perspectives on mammalian cardiovascular aging: humans to molecules SO COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY LA English DT Article; Proceedings Paper CT Annual Meeting of the Society-for-Experimental-Biology CY APR 02-03, 2001 CL UNIV KENT, CANTERBURY, ENGLAND SP Soc Exptl Biol HO UNIV KENT DE mammals; heart; vasculature; aging; stress response ID RAT CARDIAC MYOCYTES; AGE-RELATED-CHANGES; ATRIAL-NATRIURETIC-PEPTIDE; THYROID-HORMONE RECEPTOR; CHAIN GENE-EXPRESSION; OXYGEN-FREE RADICALS; RETINOID-X-RECEPTOR; GROWTH-FACTOR-BETA; SARCOPLASMIC-RETICULUM; MITOCHONDRIAL RESPIRATION AB Age-related changes in cardiovascular function and structure in healthy adult volunteer community dwelling subjects (from 20 to 85 years) is remarkable for changes in pump function [impaired left ventricular (LV) ejection reserve capacity manifest by a reduced ejection fraction and accompanied by diminished cardioacceleration, LV dilation at end diastole and an altered diastolic filling pattern] and increased vascular afterloading. There is also evidence for a reduction in the number of cardiac myocytes with advancing age. Subcellular changes with aging (best understood in rodents) include certain regulatory factors of excitation-contraction-relaxation coupling (i.e. calcium handling), modulation by adrenergic receptor (AR) stimulation, and changes in the generation and sensitivity to the damaging effects of ROS. Coordinated changes in gene expression and/or protein function with aging result in a prolonged action potential (AP), Ca(i) transient, and contraction. L-type Ca(2+) current (I(Ca)) inactivates more slowly, and outwardly-directed K(+) currents are reduced, and likely contribute to AP-prolongation. The rate of Ca(2+) sequestration by the sarcoplasmic reticulum (SR) decreases in the senescent myocardium, in part underlying the prolonged Ca(i) transient. An age-associated reduction in transcription of the SERCA2 gene, coding for the SR Ca pump, accounts in part for a decrease in the SR pump site density. The contractile response to both beta(1)-AR and beta(2)-AR stimulation diminishes with aging due to decreased adrenergic augmentation of I(Ca), and thus the Ca(i) transient, in senescent vs. young hearts. The age-associated reduction in the postsynaptic response of myocardial cells to beta(1)-AR stimulation appears to be due to multiple changes in molecular and biochemical receptor coupling and post-receptor mechanisms. An increased basal production of ROS is paralleled by increased ROS-sensitivity, markers of chronic ROS damage and mitochondrial functional decline. Overall, these changes lead to a diminished (but not necessarily exhausted) capacity of the heart to adapt to physiological or pathological stress with advancing. age. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NIA, Intramural Res Program, Cardiovasc Sci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Lakatta, EG (reprint author), NIA, Intramural Res Program, Cardiovasc Sci Lab, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. EM lakattae@grc.nia.nih.gov NR 116 TC 116 Z9 117 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1095-6433 J9 COMP BIOCHEM PHYS A JI Comp. Biochem. Physiol. A-Mol. Integr. Physiol. PD AUG PY 2002 VL 132 IS 4 BP 699 EP 721 AR PII S1095-6433(02)00124-1 DI 10.1016/S1095-6433(02)00124-1 PG 23 WC Biochemistry & Molecular Biology; Physiology; Zoology SC Biochemistry & Molecular Biology; Physiology; Zoology GA 580LG UT WOS:000177235600004 PM 12095857 ER PT J AU Ward, JM AF Ward, JM TI Comment on the muromics "garden path" SO COMPARATIVE MEDICINE LA English DT Editorial Material C1 NCI, Ft Detrick, MD 21702 USA. RP Ward, JM (reprint author), NCI, Ft Detrick, MD 21702 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC LABORATORY ANIMAL SCIENCE PI MEMPHIS PA 9190 CRESTWYN HILLS DR, MEMPHIS, TN 38125 USA SN 1532-0820 J9 COMPARATIVE MED JI Comparative Med. PD AUG PY 2002 VL 52 IS 4 BP 302 EP 302 PG 1 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 587ZJ UT WOS:000177674000001 PM 12211269 ER PT J AU Astrofsky, KM Schech, JM Sheppard, BJ Obenschain, CA Chin, AM Kacergis, MC Laver, ER Bartholomew, JL Fox, JG AF Astrofsky, KM Schech, JM Sheppard, BJ Obenschain, CA Chin, AM Kacergis, MC Laver, ER Bartholomew, JL Fox, JG TI High mortality due to Tetrahymena sp infection in laboratory-maintained zebrafish (Brachydanio rerio) SO COMPARATIVE MEDICINE LA English DT Article AB A large colony of laboratory zebrafish (Brachydanio rerio) used in the study of early vertebrate embryogenesis began experiencing acute, unexplained mortality that approached 100% among approximately 30-day-old resident fry. The initial differential diagnosis included ammonia, nitrite, or chlorine toxicosis, as well as iatrogenically induced toxicosis associated with improper sanitation procedures of laboratory equipment. Necropsy of dead and moribund fry prior to fixation revealed swarms of ovoid-shaped, motile, ciliated protozoa with a "spiraling football" motion. Wet mount preparations of various water samples also contained high numbers of similar protozoa. Histologic examination of affected fry revealed numerous, periodic acid-Schiff-positive forms within the body coelom, and epithelial and muscle tissues. The protozoa were consistent morphologically with members of the genus Tetrahymena, which is usually a free-living, nonpathogenic ciliated protozoa in fresh and saltwater environments. Relevant disease associated with Tetrahymena spp. in viviparous fish has been reported as a result of concurrent disease, immunosuppression, or poor water quality conditions. To the authors' knowledge, this is the first report of an epizootic involving laboratory maintained zebrafish, and the diagnostic course and therapeutic interventions undertaken to alleviate Tetrahymena species-associated clinical disease. C1 MIT, Div Comparat Med, Cambridge, MA 02139 USA. NICHD, NIH, Intramural Res Div, Res Anim Med Branch, Bethesda, MD 20892 USA. NICHD, NIH, Intramural Res Branch, Lab Mol Genet, Bethesda, MD 20892 USA. RP Astrofsky, KM (reprint author), MIT, Div Comparat Med, 77 Massachusetts Ave, Cambridge, MA 02139 USA. NR 15 TC 12 Z9 12 U1 1 U2 5 PU AMER ASSOC LABORATORY ANIMAL SCIENCE PI MEMPHIS PA 9190 CRESTWYN HILLS DR, MEMPHIS, TN 38125 USA SN 1532-0820 J9 COMPARATIVE MED JI Comparative Med. PD AUG PY 2002 VL 52 IS 4 BP 363 EP 367 PG 5 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 587ZJ UT WOS:000177674000015 PM 12211282 ER PT J AU Freidlin, B Kom, EL AF Freidlin, B Kom, EL TI A comment on futility monitoring SO CONTROLLED CLINICAL TRIALS LA English DT Article DE conditional power; clinical trial; futility; interim monitoring; monitoring guidelines; stochastic curtailment ID RANDOMIZED CLINICAL-TRIALS; SEQUENTIAL TEST DESIGNS; BOUNDARIES; HYPOTHESIS AB Futility monitoring of randomized clinical trials is becoming widely used. In this paper we discuss several concerns associated with aggressive monitoring for lack of activity in studies comparing experimental and control treatment arms: (1) stopping for futility when the experimental arm is doing better than the control arm, (2) conditional power not being low at the time of stopping, (3) potential loss of power to detect clinically interesting differences that are smaller than the design alternative, and (4) sensitivity of the power to the departure from the proportional hazards assumption. Our investigation suggests that aggressive futility rules do not generally reduce power (relative to less aggressive rules) under incorrect design assumptions such as overstatement of the target treatment effect or mild violations of the proportional hazards assumption. On the other hand, aggressive monitoring rules may result in an early termination for futility when the experimental arm is doing better than the control arm (in some cases nontrivially better, especially when trials are designed for unrealistically large effects). Thus, aggressive monitoring rules may fail to provide sufficiently convincing evidence to influence clinical practice or to establish a standard of treatment. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NCI, Biometr Res Branch, Bethesda, MD 20892 USA. RP Freidlin, B (reprint author), NCI, Biometr Res Branch, 6130 Execut Blvd,EPN 8122,MSC7434, Bethesda, MD 20892 USA. NR 29 TC 24 Z9 24 U1 3 U2 6 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD AUG PY 2002 VL 23 IS 4 BP 355 EP 366 AR PII S0197-2456(02)00218-0 DI 10.1016/S0197-2456(02)00218-0 PG 12 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 578XG UT WOS:000177145900001 PM 12161079 ER PT J AU Panchision, DM McKay, RDG AF Panchision, DM McKay, RDG TI The control of neural stem cells by morphogenic signals SO CURRENT OPINION IN GENETICS & DEVELOPMENT LA English DT Review ID FIBROBLAST-GROWTH-FACTOR; CENTRAL-NERVOUS-SYSTEM; RADIAL GLIAL-CELLS; LOOP-HELIX GENES; PROGENITOR CELLS; CEREBRAL-CORTEX; OLIGODENDROCYTE DIFFERENTIATION; SONIC HEDGEHOG; MAMMALIAN FOREBRAIN; DEVELOPMENTAL-CHANGES AB A complex orchestration of stem-cell specification, expansion and differentiation is required for the proper development of the nervous system. Although progress has been made on the role of individual genes in each of these processes, there are still unresolved questions about how gene function translates to the dynamic assembly of cells into tissues. Recently, stem-cell biology has emerged as a bridge between the traditional fields of cell biology and developmental genetics. In addition to their potential therapeutic role, stem cells are being exploited as experimental 'logic chips' that integrate information and exhibit self-organizing properties. Recent studies provide new insights on how morphogenic signals coordinate major stem cell decisions to regulate the size, shape and cellular diversity of the nervous system. C1 Natl Inst Neurol Disorders & Stroke, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Panchision, DM (reprint author), Natl Inst Neurol Disorders & Stroke, Mol Biol Lab, NIH, 36 Convent Dr,MSC 4092, Bethesda, MD 20892 USA. NR 117 TC 145 Z9 151 U1 0 U2 6 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0959-437X J9 CURR OPIN GENET DEV JI Curr. Opin. Genet. Dev. PD AUG PY 2002 VL 12 IS 4 BP 478 EP 487 DI 10.1016/S0959-437X(02)00329-5 PG 10 WC Cell Biology; Genetics & Heredity SC Cell Biology; Genetics & Heredity GA 570JY UT WOS:000176656200017 PM 12100896 ER PT J AU Mackness, M Rossouw, J AF Mackness, M Rossouw, J TI The Heart Protection Study: death knell for the oxidation theory of atherosclerosis? SO CURRENT OPINION IN LIPIDOLOGY LA English DT Editorial Material ID ANTIOXIDANTS; ENZYMES C1 Manchester Royal Infirm, Dept Med, Manchester M13 9WL, Lancs, England. NHLBI, Womens Hlth Inst, Bethesda, MD 20892 USA. RP Mackness, M (reprint author), Manchester Royal Infirm, Dept Med, Manchester M13 9WL, Lancs, England. NR 10 TC 1 Z9 1 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0957-9672 J9 CURR OPIN LIPIDOL JI Curr. Opin. Lipidology PD AUG PY 2002 VL 13 IS 4 BP 355 EP 355 DI 10.1097/00041433-200208000-00001 PG 1 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Peripheral Vascular Disease SC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Cardiovascular System & Cardiology GA 582PM UT WOS:000177360400001 PM 12151849 ER PT J AU Yewdell, J Garcia-Sastre, A AF Yewdell, J Garcia-Sastre, A TI Influenza virus still surprises SO CURRENT OPINION IN MICROBIOLOGY LA English DT Review ID A VIRUS; NS1 PROTEIN; NUCLEAR EXPORT; GENE; HEMAGGLUTININS; RECOGNITION; GENERATION; CELLS; DNA AB Influenza virus remains a major public heath concern, both in its annual toll in death and debilitation and its potential to cause devastating pandemics. A number of recent surprising discoveries emphasize how little we know about influenza virus and its interaction with its hosts. These include the description of a novel viral protein encoded by an overlapping reading frame, the demonstration that the most abundant viral non-structural protein interferes with the induction of interferons by infected cells, and the finding that natural killer cells express activating receptors that detect viral cell-surface proteins. The introduction of improved methods for genetically manipulating influenza virus promises to revolutionize our understanding of viral replication and its interaction with the host innate and acquired immune systems, and will also enable the improvement of vaccines. Using knowledge of viral sequences recovered from archived or interred tissues from victims of the 1918 influenza pandemic, it is now possible to investigate why this virus was so pathogenic - it killed more than 20 000 000 people, most of whom were young adults in the prime of their lives. C1 NIAID, Viral Dis Lab, Bethesda, MD 20892 USA. CUNY Mt Sinai Sch Med, Dept Microbiol, New York, NY 10029 USA. RP Yewdell, J (reprint author), NIAID, Viral Dis Lab, Rm 211,Bldg 4,4 Ctr Dr, Bethesda, MD 20892 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012; OI Garcia-Sastre, Adolfo/0000-0002-6551-1827 NR 27 TC 37 Z9 37 U1 0 U2 2 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 1369-5274 J9 CURR OPIN MICROBIOL JI Curr. Opin. Microbiol. PD AUG PY 2002 VL 5 IS 4 BP 414 EP 418 DI 10.1016/S1369-5274(02)00346-6 PG 5 WC Microbiology SC Microbiology GA 588RK UT WOS:000177714300011 PM 12160862 ER PT J AU Di Marzo, V Blumberg, PM Szallasi, A AF Di Marzo, V Blumberg, PM Szallasi, A TI Endovanilloid signaling in pain SO CURRENT OPINION IN NEUROBIOLOGY LA English DT Article ID ROOT GANGLION NEURONS; VANILLOID VR1 RECEPTORS; ACTIVATED ION-CHANNEL; PROTEIN-KINASE-C; CAPSAICIN-RECEPTOR; POSSIBLE MECHANISM; SENSORY NEURONS; MESSENGER-RNA; ANANDAMIDE; RAT AB Recent work has addressed the role of vanilloid receptor type 1 (VR1) in pain perception. VR1 activity is regulated both directly and indirectly by endogenous factors. For example, protein kinase C sensitizes human VR1 to mild decreases in pH, which are commonly encountered during inflammation, and renders the endocannabinoid anandamide a more potent 'endovanilloid'. Bradykinin and nerve growth factor release VR1 from the inhibitory control of phosphatidylinositol (4,5)-bisphosphate and anti-VR1 serum ameliorates thermal allodynia and hyperalgesia in diabetic mice. There is strong evidence that not only the sensitivity but also the density of expression of VR1 is enhanced during inflammatory conditions. These observations provide an empirical foundation which could explain the reduced inflammatory hyperalgesia in VR1 knockout mice, and they imply an important role for endovanilloid signaling via VR1 in the development of ongoing pain in humans that occurs in most inflammatory conditions. Conversely, downregulation of VR1 expression and/or activity is a promising therapeutic strategy for novel analgesic drugs. C1 Washington Univ, Sch Med, Dept Pathol & Immunol, St Louis, MO 63110 USA. NCI, Bethesda, MD 20892 USA. CNR, Ist Chim Biomol, I-80078 Naples, Italy. RP Szallasi, A (reprint author), Washington Univ, Sch Med, Dept Pathol & Immunol, Box 8118,660 S Euclid Ave, St Louis, MO 63110 USA. OI Di Marzo, Vincenzo/0000-0002-1490-3070 NR 56 TC 187 Z9 192 U1 0 U2 4 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0959-4388 J9 CURR OPIN NEUROBIOL JI Curr. Opin. Neurobiol. PD AUG PY 2002 VL 12 IS 4 BP 372 EP 379 DI 10.1016/S0959-4388(02)00340-9 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 584KL UT WOS:000177465700004 PM 12139983 ER PT J AU Weinstein, JN AF Weinstein, JN TI 'Omic' and hypothesis-driven research in the molecular pharmacology of cancer SO CURRENT OPINION IN PHARMACOLOGY LA English DT Article ID MICROARRAY; INFORMATION AB Given the layered, evolutionary complexity of biological systems, it will not be possible to understand them comprehensively on the basis of hypothesis-driven research alone. Likewise, it will not be possible to do so solely through 'omic' studies of genes, proteins and other molecules in aggregate. The two modes of research are complementary and synergistic. The twin complexities of cancer biology and drug discovery demand such synergy as a basis for the next generation of oncopharmacomic research. C1 NCI, Genom & Bioinformat Grp, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA. RP Weinstein, JN (reprint author), NCI, Genom & Bioinformat Grp, Mol Pharmacol Lab, NIH, Bldg 37 Room 5068,9000 Rockville Pike, Bethesda, MD 20892 USA. EM jw4i@nih.gov NR 18 TC 40 Z9 44 U1 1 U2 2 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4892 J9 CURR OPIN PHARMACOL JI Curr. Opin. Pharmacol. PD AUG PY 2002 VL 2 IS 4 BP 361 EP 365 DI 10.1016/S1471-4892(02)00185-6 PG 5 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 573PJ UT WOS:000176839800002 PM 12127867 ER PT J AU Holland, SM Vizi, ES AF Holland, SM Vizi, ES TI Immunomodulation SO CURRENT OPINION IN PHARMACOLOGY LA English DT Editorial Material ID IMMUNE-SYSTEM; BRAIN C1 NIAID, Immunopathogenesis Unit, Clin Pathophysiol Sect, Lab Host Def,NIH, Bethesda, MD 20892 USA. Hungarian Acad Sci, Inst Expt Sci, Dept Pharmacol, H-1083 Budapest, Hungary. RP Holland, SM (reprint author), NIAID, Immunopathogenesis Unit, Clin Pathophysiol Sect, Lab Host Def,NIH, Bldg 10,10 Ctr Dr, Bethesda, MD 20892 USA. NR 8 TC 2 Z9 2 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4892 J9 CURR OPIN PHARMACOL JI Curr. Opin. Pharmacol. PD AUG PY 2002 VL 2 IS 4 BP 425 EP 427 DI 10.1016/S1471-4892(02)00188-1 PG 3 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 573PJ UT WOS:000176839800010 PM 12127875 ER PT J AU Pau, AK Tavel, JA AF Pau, AK Tavel, JA TI Therapeutic use of interleukin-2 in HIV-infected patients SO CURRENT OPINION IN PHARMACOLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; DOSE SUBCUTANEOUS INTERLEUKIN-2; PNEUMOCYSTIS-CARINII PNEUMONIA; ACTIVE ANTIRETROVIRAL THERAPY; RANDOMIZED CONTROLLED TRIAL; PHASE-II TRIAL; INTRAVENOUS INTERLEUKIN-2; TYPE-1; DISCONTINUATION; PROPHYLAXIS AB Highly active antiretroviral therapy has improved the morbidity and survival of patients with the human immunodeficiency virus (HIV) infection. Currently available antiretroviral agents can suppress viral replication and partially reverse cellular immunity defects in HIV patients. Immune-based therapy with interleukin-2 when used as adjunctive therapy to antiretroviral therapy may further improve immune responses, as demonstrated by an increase in CD4(+) T-lymphocyte counts in recent clinical trials. C1 NIH, Ctr Clin, Dept Pharm, Bethesda, MD 20892 USA. NIAID, NIH, Bethesda, MD 20892 USA. RP Pau, AK (reprint author), NIH, Ctr Clin, Dept Pharm, Bldg 10,Room 1N257, Bethesda, MD 20892 USA. NR 33 TC 12 Z9 15 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4892 J9 CURR OPIN PHARMACOL JI Curr. Opin. Pharmacol. PD AUG PY 2002 VL 2 IS 4 BP 433 EP 439 DI 10.1016/S1471-4892(02)00177-7 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 573PJ UT WOS:000176839800012 PM 12127877 ER PT J AU Abo-Zena, RA Horwitz, ME AF Abo-Zena, RA Horwitz, ME TI Immunomodulation in stem-cell transplantation SO CURRENT OPINION IN PHARMACOLOGY LA English DT Article ID VERSUS-HOST DISEASE; BONE-MARROW TRANSPLANTATION; STEROID-REFRACTORY ACUTE; ACUTE GRAFT; MYCOPHENOLATE-MOFETIL; EXTRACORPOREAL PHOTOCHEMOTHERAPY; RANDOMIZED TRIAL; PROPHYLAXIS; CYCLOSPORINE; THERAPY AB Acute graft-versus-host disease is a complication that affects 30-60% of patients undergoing allogeneic stem-cell transplantation. The standard for prophylaxis for graft-versus-host disease has historically been the combination of cyclosporine and methotrexate. Recently, tacrolimus has been used more frequently and current studies are exploring the potential of mycophenolate mofetil. There is little published experience with the use of sirolimus in prophylaxis or treatment but studies are ongoing. There have been significant advances recently in the treatment of steroid-refractory acute graft-versus-host disease. Historically, antithymocyte globulin was used when patients did not respond to the steroid treatment. New monoclonal antibodies such as daclizumab, and tumor necrosis factor alpha inhibitors such as infliximab are producing more promising results. Chronic graft-versus-host disease continues to be a major complication of stem-cell transplantation, affecting 35-50% of patients. Finding effective treatments for chronic graft-versus-host disease other than steroids continues to be a challenge. C1 NIH, Dept Pharm, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Abo-Zena, RA (reprint author), NIH, Dept Pharm, Bldg 10,Room 1N-257, Bethesda, MD 20892 USA. NR 45 TC 7 Z9 7 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4892 J9 CURR OPIN PHARMACOL JI Curr. Opin. Pharmacol. PD AUG PY 2002 VL 2 IS 4 BP 452 EP 457 DI 10.1016/S1471-4892(02)00174-1 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 573PJ UT WOS:000176839800015 PM 12127880 ER PT J AU Chanock, S Taylor, JG AF Chanock, S Taylor, JG TI Using genetic variation to study immunomodulation SO CURRENT OPINION IN PHARMACOLOGY LA English DT Article ID MANNOSE-BINDING LECTIN; SINGLE-NUCLEOTIDE POLYMORPHISMS; SYSTEMIC-LUPUS-ERYTHEMATOSUS; THIOPURINE S-METHYLTRANSFERASE; CHRONIC GRANULOMATOUS-DISEASE; CYSTIC-FIBROSIS; PROTEIN GENE; POPULATION STRATIFICATION; RHEUMATOID-ARTHRITIS; VARIANT ALLELES AB The generation of a draft sequence of a human genome has led to the identification of millions of common variants, known as single nucleotide polymorphisms, which constitute a resource for studying complex diseases. Currently, high-density maps of variants in candidate genes, chromosomal regions or the entire genome should encourage investigation of determinants of human immune response, using quantitative analysis. Ultimately, this approach should identify novel targets for therapeutic intervention. C1 NCI, Sect Genom Variat, Pediat Oncol Branch, Ctr Adv Technol, Gaithersburg, MD 20877 USA. RP Chanock, S (reprint author), NCI, Sect Genom Variat, Pediat Oncol Branch, Ctr Adv Technol, 8717 Grovemont Circle, Gaithersburg, MD 20877 USA. OI Taylor, James/0000-0002-4421-1809 NR 64 TC 9 Z9 9 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4892 J9 CURR OPIN PHARMACOL JI Curr. Opin. Pharmacol. PD AUG PY 2002 VL 2 IS 4 BP 463 EP 469 DI 10.1016/S1471-4892(02)00186-8 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 573PJ UT WOS:000176839800017 PM 12127882 ER PT J AU Frankel, AE Powell, BL Duesbery, NS Woude, GFV Leppla, SH AF Frankel, AE Powell, BL Duesbery, NS Woude, GFV Leppla, SH TI Anthrax fusion protein therapy of cancer SO CURRENT PROTEIN & PEPTIDE SCIENCE LA English DT Review ID TOXIN LETHAL FACTOR; COLONY-STIMULATING FACTOR; DEPENDENT ADENYLATE-CYCLASE; RECEPTOR-BINDING REGION; ACUTE MYELOID-LEUKEMIA; PHASE-I TRIAL; BACILLUS-ANTHRACIS; PROTECTIVE ANTIGEN; MAMMALIAN-CELLS; PSEUDOMONAS EXOTOXIN AB Most patients with cancer are treated with chemotherapy but die front progressive disease or toxicities of therapy. Current chemotherapy regimens primarily use cytotoxic drugs which damage cell DNA or impair cell proliferation in both malignant and normal tissues. After several treatment courses, the patients' tumor cells often overexpress multi-drug resistance genes which prevent further tumor cytoreduction. Novel agents which can kill such resistant tumor cells are needed. One such class of agents are targeted peptide toxins. Targeted peptide toxins consist of peptide toxins covalently linked to tumor selective peptide ligands. These molecules bind tumor cell surface receptors, internalize, and facilitate transfer of the toxin catalytic domains to the cytosol. Once in the cytosol, the enzyme activity leads to cell death. A number of plant, bacterial and fungal toxins have been used, and clinical trials with several of these have produced complete remissions in chemoresistant neoplasms. Nevertheless, there is a continuing need for navel targeted toxins. Many patients have pre-existing antibodies against the currently clinically used toxins and many toxins are inactive when used for myeloid malignancies where internalized proteins are rapidly routed and degraded in lysosomes. Anthrax toxins are the cytotoxic components of Bacillus anthracis. While the bacteria has been the source of serious illness, deaths and global anxieties related to past or future bioterrorism, the isolated toxins do not pose public health hazards. In fact, toxin treated patients will likely develop protective antibodies. Anthrax toxin is an excellent choice for tumor cell surface targeting. Other than U.S. military personnel immunized during the Gulf War, most people lack preexisting antibodies. This may change in the future due to threats of additional terrorist acts, but for the present few patients will have antibodies to anthrax proteins. The separate subunits for binding, translocation and cell killing facilitate genetic engineering to yield tumor-specific cell killing. The toxins are more potent than most of the other peptide toxins and may yield highly efficacious targeted molecules. This essay will review anthrax toxin structure-function, preliminary experiments with re-targeted anthrax toxin and potential designs for new ligand-anthrax therapeutics. C1 Wake Forest Univ, Bowman Gray Sch Med, Dept Med, Winston Salem, NC 27103 USA. Van Andel Inst, Grand Rapids, MI USA. NIDCR, Oral Infect & Immun Branch, Bethesda, MD USA. RP Frankel, AE (reprint author), Wake Forest Univ, Bowman Gray Sch Med, Dept Med, Winston Salem, NC 27103 USA. OI DUESBERY, NICK/0000-0002-4258-5655 NR 75 TC 17 Z9 18 U1 0 U2 3 PU BENTHAM SCIENCE PUBL LTD PI HILVERSUM PA PO BOX 1673, 1200 BR HILVERSUM, NETHERLANDS SN 1389-2037 J9 CURR PROTEIN PEPT SC JI Curr. Protein Pept. Sci. PD AUG PY 2002 VL 3 IS 4 BP 399 EP 407 DI 10.2174/1389203023380567 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 572RJ UT WOS:000176788500002 PM 12370003 ER PT J AU Frisch, M AF Frisch, M TI On the etiology of anal squamous carcinoma SO DANISH MEDICAL BULLETIN LA English DT Review ID HUMAN-PAPILLOMAVIRUS INFECTION; HUMAN-IMMUNODEFICIENCY-VIRUS; CERVICAL INTRAEPITHELIAL NEOPLASIA; SEXUALLY-TRANSMITTED DISEASES; INFLAMMATORY-BOWEL-DISEASE; RENAL-TRANSPLANT PATIENTS; NON-HODGKINS-LYMPHOMA; 2ND PRIMARY CANCERS; HOMOSEXUAL MEN; CROHNS-DISEASE C1 Statens Serum Inst, Dept Epidemiol Res, DK-2300 Copenhagen S, Denmark. Natl Canc Inst, Bethesda, MD USA. Danish Canc Registry, Copenhagen, Denmark. RP Frisch, M (reprint author), Statens Serum Inst, Dept Epidemiol Res, Artillerivej 3, DK-2300 Copenhagen S, Denmark. RI Frisch, Morten/E-9206-2016 OI Frisch, Morten/0000-0002-3864-8860 NR 152 TC 50 Z9 59 U1 0 U2 3 PU DANISH MEDICAL ASSN PI COPENHAGEN PA TRONDHJEMSGADE 9, DK-2100 COPENHAGEN, DENMARK SN 0907-8916 J9 DAN MED BULL JI Dan. Med. Bull. PD AUG PY 2002 VL 49 IS 3 BP 194 EP 209 PG 16 WC Medicine, General & Internal SC General & Internal Medicine GA 595XE UT WOS:000178133200002 PM 12238281 ER PT J AU Cronk, KM Wilkinson, GA Grimes, R Wheeler, EF Jhaveri, S Fundin, BT Silos-Santiago, I Tessarollo, L Reichardt, LF Rice, FL AF Cronk, KM Wilkinson, GA Grimes, R Wheeler, EF Jhaveri, S Fundin, BT Silos-Santiago, I Tessarollo, L Reichardt, LF Rice, FL TI Diverse dependencies of developing Merkel innervation on the trkA and both full-length and truncated isoforms of trkC SO DEVELOPMENT LA English DT Article DE neurotrophins; cutaneous innervation; trigeminal; mechanoreceptors; mouse ID DIFFERENTIAL DEPENDENCY; NEURONAL DEVELOPMENT; NONNEURONAL CELLS; SENSORY NEURONS; MYSTACIAL PAD; RECEPTOR GENE; RAT TRKC; IN-VIVO; NEUROTROPHIN-3; EXPRESSION AB This study demonstrates that innervation dependent on two different neurotrophin tyrosine kinase (trk) receptors can form the same types of sensory endings (Merkel endings) in the same target (Merkel cells of vibrissa follicles). Some endings transiently express trkA during their initial development, whereas others express trkC throughout their development. Consequently, elimination of kinase domains of either trkA or trkC each result in a partial loss of Merkel endings, whereas absence of kinase domains of both receptors results in a total loss. At the onset of Merkel ending development, at least one kinase-lacking trkC isoform is transiently expressed on all the follicle cells, while neurotrophin 3 is transiently expressed only in the cells at the middle third of the follicle where the Merkel endings and cells develop. This transient non-neuronal expression of truncated trkC is essential for development of any Merkel endings, whereas some Merkel endings and cells still begin to develop in the absence of neurotrophin 3. Therefore, truncated trkC plays a more important role in the development of this innervation than kinase forms of trkA or trkC or of NT3, the only known ligand for trkC receptors. C1 Albany Med Coll, Ctr Neuropharmacol & Neurosci, Albany, NY 12208 USA. Univ Calif San Francisco, Howard Hughes Med Ctr, Neurosci Unit, San Francisco, CA 94143 USA. Univ Texas, Div Life Sci, San Antonio, TX 78249 USA. MIT, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA. Astra Zeneca R&D Sodertalje, Dept Mol Sci, SE-14157 Huddinge, Sweden. Millennium Pharmaceut Inc, Dept Neurobiol, Cambridge, MA 02139 USA. NCI, FCRDC, Neural Dev Grp, Frederick, MD 21702 USA. RP Rice, FL (reprint author), Albany Med Coll, Ctr Neuropharmacol & Neurosci, Albany, NY 12208 USA. FU NEI NIH HHS [T32 EY 07120, T32 EY007120]; NIGMS NIH HHS [GM08194, S06 GM008194]; NINDS NIH HHS [P01 NS 16033, P01 NS016033-19, R01 NS34692, P01 NS016033] NR 40 TC 34 Z9 35 U1 1 U2 1 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD AUG PY 2002 VL 129 IS 15 BP 3739 EP 3750 PG 12 WC Developmental Biology SC Developmental Biology GA 586EC UT WOS:000177570800019 PM 12117822 ER PT J AU Brody, T Odenwald, WF AF Brody, T Odenwald, WF TI Cellular diversity in the developing nervous system: a temporal view from Drosophila SO DEVELOPMENT LA English DT Review DE Drosophila; nervous system; neural precursor cell ID POU DOMAIN GENES; CEREBRAL CORTICAL DEVELOPMENT; NEURAL STEM-CELLS; TRANSCRIPTION FACTORS; NEUROBLAST LINEAGES; RETINAL DEVELOPMENT; INDUCTIVE SIGNALS; DORSOVENTRAL AXIS; PATTERN-FORMATION; EXPRESSION AB This article considers the evidence for temporal transitions in CNS neural precursor cell gene expression during development. In Drosophila, five prospective competence states have so far been identified, characterized by the successive expression of Hb-->Kr-->Pdm-->Cas-->Gh in many, but not all, neuroblasts. In each temporal window of transcription factor expression, the neuroblast generates sublineages whose temporal identity is determined by the competence state of the neuroblast at the time of birth of the sublineage. Although similar regulatory programs have not yet been identified in mammals, candidate regulatory genes have been identified. Further investigation of the genetic programs that guide both invertebrate and vertebrate neural precursor cell lineage development will ultimately lead to an understanding of the molecular events that control neuronal diversity. C1 NINDS, Neurogenet Unit, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP NINDS, Neurogenet Unit, Neurochem Lab, NIH, Bldg 36,Rm 4D04, Bethesda, MD 20892 USA. EM ward@codon.nih.gov NR 82 TC 45 Z9 45 U1 2 U2 3 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 EI 1477-9129 J9 DEVELOPMENT JI Development PD AUG PY 2002 VL 129 IS 16 BP 3763 EP 3770 AR UNSP DEV9854 PG 8 WC Developmental Biology SC Developmental Biology GA 591EV UT WOS:000177867500001 PM 12135915 ER PT J AU Kawahara, A Chien, CB Dawid, IB AF Kawahara, A Chien, CB Dawid, IB TI The homeobox gene mbx is involved in eye and tectum development SO DEVELOPMENTAL BIOLOGY LA English DT Article DE mbx; homeobox gene; eye; tectum; midbrain-hindbrain boundary; morpholino ID MIDBRAIN-HINDBRAIN BOUNDARY; RETINOTECTAL PROJECTION; CEREBELLUM DEVELOPMENT; NEURAL PLATE; DANIO-RERIO; ZEBRAFISH; EXPRESSION; MUTATIONS; ORGANIZER; PAX7 AB The homeobox gene mbx is first activated at the end of gastrulation in zebrafish in the presumptive forebrain and midbrain region. During somitogenesis stages, the anterior expression of mbx, which partly overlaps the future eye field, gradually decreases, while midbrain expression intensifies and becomes restricted to the presumptive rectum. Knockdown of mbx expression by morpholino antisense oligonucleotides (mbx-MO) leads to a reduction in the size of the eyes and tectum. Expression domains of rx1 and pax6 in the eye field and of mab21/2 in the eye field and tectum anlage were reduced in size in mbx-MO-injected embryos by somitogenesis stages. Further, induction of islet1 and lim3 expression in the eye at 2 days postfertilization (dpf) was suppressed in mbx-MO-injected embryos. In mbx-MO-injected embryos at 2-5 dpf, the lamination of the eye was disorganized and the number of retinal axons was substantially reduced, but the few remaining axons navigated appropriately to the contralateral tectum. A chimeric protein composed of the Mbx DNA-binding domain and the VP16 activation domain affected eye and rectum development similarly to mbx-MO knockdown, suggesting that Mbx acts as a transcriptional repressor in the zebrafish embryo. Based on these data, we propose that the mbx homeobox gene is required for the development of the eyes and tectum. C1 NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. Univ Utah, Med Ctr, Dept Neurobiol & Anat, Salt Lake City, UT 84132 USA. RP Kawahara, A (reprint author), NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. NR 43 TC 49 Z9 53 U1 1 U2 2 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD AUG 1 PY 2002 VL 248 IS 1 BP 107 EP 117 DI 10.1006/dbio.2002.0709 PG 11 WC Developmental Biology SC Developmental Biology GA 580WG UT WOS:000177259200009 PM 12142024 ER PT J AU Hanley, AJG Karter, AJ Festa, A D'Agostino, R Wagenknecht, LE Savage, P Tracy, RP Saad, MF Haffner, S AF Hanley, AJG Karter, AJ Festa, A D'Agostino, R Wagenknecht, LE Savage, P Tracy, RP Saad, MF Haffner, S TI Factor analysis of metabolic syndrome using directly measured insulin sensitivity - The Insulin Resistance Atherosclerosis Study SO DIABETES LA English DT Article ID CARDIOVASCULAR RISK-FACTORS; CORONARY HEART-DISEASE; GLUCOSE-TOLERANCE TEST; MINIMAL MODEL; DIABETES-MELLITUS; SYNDROME PREDICTS; SYNDROME-X; MEN; INFLAMMATION; POPULATION AB Factor analysis, a multivariate correlation technique, has been used to provide insight into the underlying structure of metabolic syndrome, which is characterized by physiological complexity and strong statistical intercorrelation among its key variables. The majority of previous factor analyses, however, have used only surrogate measures of insulin sensitivity. In addition, few have included members of multiple ethnic groups, and only one has presented results separately for subjects with impaired glucose tolerance. The objective of this study was to investigate, using factor analysis, the clustering of physiologic variables using data from 1,087 nondiabetic participants in the Insulin Resistance Atherosclerosis Study (IRAS). This study includes information on the directly measured insulin sensitivity index (SI) from intravenous glucose tolerance testing among African-American, Hispanic, and non-Hispanic white subjects aged 40-69 years at various stages of glucose tolerance. Principal factor analysis identified two factors that explained 28 and 9% of the variance in the dataset, respectively. These factors were interpreted as 1) a "metabolic" factor, with positive loadings of BMI, waist, fasting and 2-h glucose, and triglyceride and inverse loadings of log(S-I + 1) and HDL; and 2) a "blood pressure" factor, with positive loadings of systolic and diastolic blood pressure. The results were unchanged when surrogate measures of insulin resistance were used in place of log(S-I + 1). In addition, the results were similar within strata of sex, glucose tolerance status, and ethnicity. In conclusion, factor analysis identified two underlying factors among a group of metabolic syndrome variables in this dataset. Analyses using surrogate measures of insulin resistance suggested that these variables provide adequate information to explore the underlying intercorrelational structure of metabolic syndrome. Additional clarification of the physiologic characteristics of metabolic syndrome is required as individuals with this condition are increasingly being considered candidates for behavioral and pharmacologic intervention. C1 Univ Texas, Hlth Sci Ctr, Div Clin Epidemiol, San Antonio, TX 78229 USA. Mt Sinai Hosp, Samuel Lunenfeld Res Inst, Div Epidemiol & Biostat, Toronto, ON M5G 1X5, Canada. Kaiser Permanente, Div Res, Oakland, CA USA. Wake Forest Univ, Sch Med, Dept Publ Hlth Sci, Winston Salem, NC 27109 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Univ Vermont, Coll Med, Dept Pathol, Burlington, VT 05405 USA. Univ Vermont, Coll Med, Dept Biochem, Burlington, VT 05405 USA. Univ S Carolina, Dept Med, Sch Med, Los Angeles, CA USA. RP Haffner, S (reprint author), Univ Texas, Hlth Sci Ctr, Div Clin Epidemiol, 7703 Floyd Curl Dr,Mail Code 7873, San Antonio, TX 78229 USA. RI Dagostino Jr, Ralph/C-4060-2017 OI Dagostino Jr, Ralph/0000-0002-3550-8395 FU NHLBI NIH HHS [U01-HL47889, U01-HL47887, U01-HL47892, U01-HL47902]; NIDDK NIH HHS [DK-29867]; PHS HHS [R01 58329] NR 38 TC 133 Z9 151 U1 0 U2 4 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD AUG PY 2002 VL 51 IS 8 BP 2642 EP 2647 DI 10.2337/diabetes.51.8.2642 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 579NZ UT WOS:000177185900040 PM 12145182 ER PT J AU Meigs, JB Larson, MG D'Agostino, RB Levy, D Clouse, ME Nathan, DM Wilson, PWF O'Donnell, CJ AF Meigs, JB Larson, MG D'Agostino, RB Levy, D Clouse, ME Nathan, DM Wilson, PWF O'Donnell, CJ TI Coronary artery calcification in type 2 diabetes and insulin resistance - The Framingham Offspring Study SO DIABETES CARE LA English DT Article; Proceedings Paper CT American-Heart-Association Annual Conference on Cardiovascular Disease Epidemiology and Prevention CY MAR 02, 2001 CL SAN ANTONIO, TEXAS SP Amer Heart Assoc ID BEAM COMPUTED-TOMOGRAPHY; PREDICT CARDIOVASCULAR RISK; INTIMAL-MEDIAL THICKNESS; CAROTID WALL THICKNESS; HEART-DISEASE; GLUCOSE-TOLERANCE; MYOCARDIAL-INFARCTION; NONDIABETIC SUBJECTS; PLASMA-GLUCOSE; ATHEROSCLEROSIS AB OBJECTIVE- To assess risk for subclinical coronary atherosclerosis using electron beam-computed tomography in subjects with or without insulin resistance and with normal glucose tolerance (NGT) or impaired glucose tolerance (IGT/impaired fasting glucose [IFG]) or type 2 diabetes. RESEARCH DESIGN AND METHODS- We categorized glucose tolerance by type 2 diabetes therapy (diagnosed diabetes) or with an oral glucose tolerance test (OGTT) (IFG, IGT, and OGTT-detected diabetes) and insulin resistance as an elevated fasting insulin level, in subjects attending the fifth examination (1991-1995) of the Framingham Offspring Study. A representative subset of subjects without clinical atherosclerosis was selected for electron beam computed tomography in 1998-1999 from age- and sex-stratified quintiles of the Framingham risk score. The presence of subclinical atherosclerosis was defined as the upper quartile of the Agatston score distribution (score > 170). We assessed risk for subclinical atherosclerosis using multivariable logistic regression. RESULTS- Of 325 subjects aged 31-73 years, 51% were men, 11.2% had IFG/IGT, and 9.9% had diabetes (2.8% with diagnosed diabetes); 14.5% had insulin resistance. Compared with NGT, subjects with IFG/IGT tended to be more likely (adjusted odds ratio 1.5, 95% CI 0.7-3.4) and those with diabetes were significantly more likely (2.7, 1.2-6.1) to have subclinical coronary atherosclerosis. In age- and sex-adjusted models, subjects with insulin resistance were more likely to have subclinical atherosclerosis than those without insulin resistance (2.1, 1.01-4.2), but further risk factor adjustment weakened this association. In adjusted models including insulin resistance, diabetes remained associated with risk for subclinical atherosclerosis (2.8, 1.2-6.7); diagnosed diabetes (6.0, 1.4-25.2) had a larger effect than OGTT-detected diabetes (2.1, 0.8-5.5). CONCLUSIONS- Individuals with diabetes have an elevated burden of subclinical coronary atherosclerosis. Aggressive clinical atherosclerosis prevention is warranted, especially in diagnosed diabetes. C1 Massachusetts Gen Hosp, Div Gen Med, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Med, Boston, MA 02114 USA. Harvard Univ, Sch Med, Boston, MA 02115 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Dept Math Stat, Boston, MA 02215 USA. Boston Univ, Consulting Unit, Boston, MA 02215 USA. Beth Israel Deaconess Med Ctr, Dept Radiol, Boston, MA 02215 USA. Massachusetts Gen Hosp, Diabet Unit, Boston, MA 02114 USA. Boston Univ, Sch Med, Framingham Heart Study, Framingham, MA USA. Massachusetts Gen Hosp, Div Cardiol, Boston, MA 02114 USA. RP Meigs, JB (reprint author), Massachusetts Gen Hosp, Div Gen Med, 50 Staniford St,9th Floor, Boston, MA 02114 USA. FU NHLBI NIH HHS [N01-HC-38083] NR 43 TC 74 Z9 75 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1701 N BEAUREGARD ST, ALEXANDRIA, VA 22311-1717 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD AUG PY 2002 VL 25 IS 8 BP 1313 EP 1319 DI 10.2337/diacare.25.8.1313 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 724UB UT WOS:000185504100008 PM 12145227 ER PT J AU Wang, WY Lee, ET Fabsitz, R Welty, TK Howard, BV AF Wang, WY Lee, ET Fabsitz, R Welty, TK Howard, BV TI Using HbA(1c) to improve efficacy of the American Diabetes Association fasting plasma glucose criterion in screening for new type 2 diabetes in American Indians - The Strong Heart Study SO DIABETES CARE LA English DT Article ID DIAGNOSTIC-CRITERIA; GLYCATED HEMOGLOBIN; MELLITUS; INSULIN; CLASSIFICATION; TOLERANCE; TESTS; COMPLICATIONS; POPULATION; LOAD AB OBJECTIVE - To find an optimal critical line in the fasting plasma glucose (FPG)-HbA(1c) plane for identifying diabetes in participants with impaired fasting glucose (IFG) and thereby improve the efficacy of using FPG alone in diabetes screening among American Indians. RESEARCH DESIGN AND METHODS - We used FPG, 2-h postload glucose (2hPG), and HbA(1c) measured in the 2,389 American Indians (aged 45-74 years, without diabetes treatment or prior history of diabetes) in the Strong Heart Study (SHS) baseline (second) examination. Participants were classified as having diabetes if they had either FPG greater than or equal to 126 mg/dl or 2hPG greater than or equal to200 mg/dl, as having IFG if they had 110 less than or equal to FPG < 126 mg/dl, and as having normal fasting glucose (NFG) if they had FPG <110, according to the American Diabetes Association (ADA) definition. Logistic regression models were used for identifying diabetes (2hPG greater than or equal to200 mg/dl) in IFG participants. The areas under the receiver operating characteristic (ROC) curves generated by different logistic regression models were evaluated and compared to select the best model. A utility function based on the best model and the cost-to-benefit ratio was used to find the optimal critical line. The data from the second examination were used to study the effect of the time. I interval between the successive diabetes screenings on both the FPG criterion and the optimal critical tine. RESULTS - A total of 37% of all subjects with new diabetes at baseline and 55.2% of those in the second exam had 2hPG greater than or equal to200 but FPG <126. There was a very large portion of IFG participants With diabetes (19.3 and 22.9% in the baseline and second exam, respectively). Among the areas under the ROC curves, the area generated by the logistic regression model on FPG plus HbA(1c) is the largest and is significantly larger than that based on FPG (P = 0.0008). For a cost-to-benefit ratio of 0.23888, the optimal critical line that has the highest utility is: 0.89 X HbA(1c) + 0.11 X FPG = 17.92. Those IFG participants whose FPG and HbA(1c) were above or on the line were referred to take an oral glucose tolerance test (OGTT) to diagnose diabetes. The optimal critical line is lower if a successive diabetes screening will be conducted 4 years after the previous screening. CONCLUSIONS - FPG greater than or equal to126 and 2hPG greater than or equal to200, as suggested by the ADA, are used independently to define diabetes. The FPG level is easy to obtain, and using FPG alone is suggested for diabetes screening. It is difficult to get physicians and patients to perform an OGTT to get a 2hPG level because of the many drawbacks of the OGTT, especially in those patients who already have FPG <126. It is also impractical to conduct an OGTT for everyone in a diabetes screening. Our data show that 37% of all subjects with new diabetes in the SHS baseline exam and 55.2% of those in the second exam have 2hPG 200 but FPG <126. These cases of diabetes cannot be detected if FPG is used alone in a diabetes screening. Therefore, although the small portion of diabetes in the NFG group (4.7% in the baseline and 6.9% in the second exam) may be ignored, those cases of diabetes among IFG participants (similar to20% in our data) need further. consideration in a diabetes screening. It may be worthwhile for those IFG participants identified by the optimal critical line to take an OGTT. The optimal critical line and time interval between successive diabetes screenings need further study. C1 Univ Oklahoma, Coll Publ Hlth, Hlth Sci Ctr, Oklahoma City, OK 73190 USA. NHLBI, Bethesda, MD USA. Aberdeen Area Tribal Chairmens Hlth Board, Aberdeen, SD USA. Medlant Res Inst, Washington, DC USA. RP Wang, WY (reprint author), Univ Oklahoma, Coll Publ Hlth, Hlth Sci Ctr, POB 26901, Oklahoma City, OK 73190 USA. NR 27 TC 25 Z9 26 U1 0 U2 1 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1701 N BEAUREGARD ST, ALEXANDRIA, VA 22311-1717 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD AUG PY 2002 VL 25 IS 8 BP 1365 EP 1370 DI 10.2337/diacare.25.8.1365 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 724UB UT WOS:000185504100017 PM 12145236 ER PT J AU Nammo, T Yamagata, K Hamaoka, R Zhu, Q Akiyama, TE Gonzalez, FJ Miyagawa, J Matsuzawa, Y AF Nammo, T Yamagata, K Hamaoka, R Zhu, Q Akiyama, TE Gonzalez, FJ Miyagawa, J Matsuzawa, Y TI Expression profile of MODY3/HNF-1 alpha protein in the developing mouse pancreas SO DIABETOLOGIA LA English DT Article DE HNF-1 alpha; transcription factor; development; mouse pancreas ID HEPATOCYTE NUCLEAR FACTOR-1-ALPHA; INSULIN GENE-TRANSCRIPTION; BETA-CELLS; DIABETES-MELLITUS; HNF-1-ALPHA FUNCTION; ISLET DEVELOPMENT; JAPANESE SUBJECTS; FACTOR 1-ALPHA; DNA-BINDING; ALPHA-CELLS AB Aims/hypothesis. One subtype of MODY (MODY3) results from the heterozygous mutation of a hepatocyte nuclear factor (HNF)-1alpha. The pattern of HNF-1alpha expression in the normal pancreas has not been determined. This study aimed to clarify the profile of HNF-1alpha protein expression in the developing mouse pancreas. Methods. Double immunofluorescence staining was carried out for HNF-1alpha and pancreatic hormones or transcription factors (PDX-1, Pax6, Isl1, and Nkx2.2). The expression of these transcription factors was also studied in the beta cells of HNF-1alpha mutant mice. Results. HNF-1a was expressed by both endocrine and exocrine cells of the pancreas. Double immunofluorescence staining showed that HNF-1alpha was expressed in the nuclei of alpha cells, beta cells, delta cells, and pancreatic polypeptide (PP) cells. HNF-1alpha was first detected in most pancreatic epithelial cells on embryonic day 10.5 (E10.5), and hormone-positive endocrine cells and amylase-positive cells expressed HNF-1alpha on E15.5. Most of the Pax6-, Isl1-, or PDX-1-positive cells showed co-expression of HNF-1alpha. However, HNF-1alpha immunoreactivity was not observed in 36.0% of Nkx2.2-positive cells. Expression of Nkx2.2, Isl1 and Pax6 seemed to be normal in the beta cells of transgenic mice with dominant negative overexpression of HNF-1alpha. Expression of PDX-1 did not change in the beta cells of pre-diabetic HNF-1alpha (-/-) mice, but expression was markedly decreased in the diabetic stage. Conclusion/interpretation. HNF-1alpha is expressed by both endocrine cells and exocrine cells of the pancreas from the foetal stage along with other transcription factors, so HNF-1alpha might play a role during development. C1 Osaka Univ, Grad Sch Med, Dept Internal Med & Mol Sci, Suita, Osaka 5650871, Japan. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. RP Yamagata, K (reprint author), Osaka Univ, Grad Sch Med, Dept Internal Med & Mol Sci, 2-2 Yamadaoka,B5, Suita, Osaka 5650871, Japan. NR 49 TC 19 Z9 19 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 IS 8 BP 1142 EP 1153 DI 10.1007/s00125-002-0892-8 PG 12 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 591EC UT WOS:000177865900009 PM 12189445 ER PT J AU Haluzik, M Colombo, C Cutson, J Dietz, KR Gavrilova, O Reitman, ML AF Haluzik, M Colombo, C Cutson, J Dietz, KR Gavrilova, O Reitman, ML TI C57BL/6J vs. FVB/N genetic background strongly influences liver and muscle insulin sensitivity in lipoatrophic A-ZIP/F-1 mice SO DIABETOLOGIA LA English DT Meeting Abstract CT 38th EASD Annual Meeting of the European-Association-for-the-Study-of-Diabetes CY SEP 01-05, 2002 CL BUDAPEST, HUNGARY SP European Assoc Study Diabetes (EASD) C1 NIDDK, Diabet Branch, NIH, Bethesda, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 SU 2 MA 246 BP A83 EP A83 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 590HM UT WOS:000177813600247 ER PT J AU Stumvoll, M Tataranni, PA Stefan, N Vozarova, B Bogardus, C AF Stumvoll, M Tataranni, PA Stefan, N Vozarova, B Bogardus, C TI Increased glycemia physiologically accompanies insulin resistance due to incomplete beta cell compensation SO DIABETOLOGIA LA English DT Meeting Abstract CT 38th EASD Annual Meeting of the European-Association-for-the-Study-of-Diabetes CY SEP 01-05, 2002 CL BUDAPEST, HUNGARY SP European Assoc Study Diabetes (EASD) C1 NIDDK, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 SU 2 MA 570 BP A185 EP A185 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 590HM UT WOS:000177813600570 ER PT J AU Vozar, J Hanson, R De Courten, M Zahorakova, A Egyenes, HP Tataranni, A Bennett, P Vozarova, B AF Vozar, J Hanson, R De Courten, M Zahorakova, A Egyenes, HP Tataranni, A Bennett, P Vozarova, B TI Higher prevalence rates of cardiovascular disease in gypsies compared to Caucasians in Slovakia SO DIABETOLOGIA LA English DT Meeting Abstract CT 38th EASD Annual Meeting of the European-Association-for-the-Study-of-Diabetes CY SEP 01-05, 2002 CL BUDAPEST, HUNGARY SP European Assoc Study Diabetes (EASD) C1 Reg Hosp, Samorin, Slovakia. NIDDKD, NIH, Phoenix, AZ USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 SU 2 MA 283 BP A95 EP A95 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 590HM UT WOS:000177813600283 ER PT J AU Vozarova, B Fernandez-Real, J Knowler, WC Hanson, RL Ricart, W Gallart, L Tataranni, AP Wolford, JK AF Vozarova, B Fernandez-Real, J Knowler, WC Hanson, RL Ricart, W Gallart, L Tataranni, AP Wolford, JK TI The interleukin-6 (474) G/C promoter polymorphism is associated with type 2 diabetes mellitus in American Indians and Caucasians SO DIABETOLOGIA LA English DT Meeting Abstract CT 38th EASD Annual Meeting of the European-Association-for-the-Study-of-Diabetes CY SEP 01-05, 2002 CL BUDAPEST, HUNGARY SP European Assoc Study Diabetes (EASD) C1 NIDDKD, NIH, Phoenix, AZ USA. Univ Hosp Girona Dr Josep Trueta, Unitat Diabetol Endocrinol Nutr, Girona, Spain. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 SU 2 MA 160 BP A56 EP A56 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 590HM UT WOS:000177813600161 ER PT J AU Zierath, JR Bjornholm, M Attersand, A Lake, S He, AR Liu, SCH Lienhard, GE Taylor, S Arner, P AF Zierath, JR Bjornholm, M Attersand, A Lake, S He, AR Liu, SCH Lienhard, GE Taylor, S Arner, P TI Absence of functional insulin receptor substrate-3 (IRS-3) gene in humans SO DIABETOLOGIA LA English DT Meeting Abstract CT 38th EASD Annual Meeting of the European-Association-for-the-Study-of-Diabetes CY SEP 01-05, 2002 CL BUDAPEST, HUNGARY SP European Assoc Study Diabetes (EASD) C1 Karolinska Inst, Dept Clin Physiol, S-10521 Stockholm, Sweden. Karolinska Inst, Dept Physiol & Pharmacol, S-10521 Stockholm, Sweden. Biovitrum, Dept Biol, Stockholm, Sweden. NIDDKD, NIH, Bethesda, MD 20892 USA. Dartmouth Coll, Hitchcock Med Ctr, Dartmouth Med Sch, Dept Biochem, Hanover, NH 03756 USA. Lilly Res Labs, Indianapolis, IN USA. Huddinge Univ Hosp, Med & Clin Res Ctr, S-14186 Huddinge, Sweden. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD AUG PY 2002 VL 45 SU 2 MA 130 BP A45 EP A46 PG 2 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 590HM UT WOS:000177813600131 ER PT J AU Horikawa, I Barrett, JC AF Horikawa, I Barrett, JC TI cDNA cloning of the human polybromo-1 gene on chromosome 3p21 SO DNA SEQUENCE LA English DT Article DE bromodomain; chromosome 3p21; hPB1 gene; polybromo-1 gene ID REGION; BROMODOMAIN; COOPERATE; PROTEIN; COMPLEX; CANCER AB The bromodomain is suggested as a chromatin-targeting domain involved in regulation of gene expression and chromatin structure. We here report the cDNA cloning of the human polybromo-1 (hPB1) gene that encodes a 1634-amino acid polypeptide containing six bromodomains. The hPB1 polypeptide shows 91% identity to the previously identified chicken polybromo-1 with conservation of the bromodomains and other characteristic features. Northern blot analysis detected the ubiquitous expression of hPB1 mRNA in a variety of human tissues. Four alternative splicings were found within the hPB1 coding region: three making in-frame deletions and one resulting in a C-terminal truncation. The hPB1 gene is located on chromosome 3p21, where the tumor suppressor genes for breast, lung and kidney cancers have been mapped. C1 NCI, Lab Biosyst & Canc, Ctr Canc Res, Bethesda, MD 20892 USA. RP Horikawa, I (reprint author), NCI, Lab Biosyst & Canc, Ctr Canc Res, 9000 Rockville Pike,Bldg 37,Rm 5046,MSC-4264, Bethesda, MD 20892 USA. NR 16 TC 4 Z9 5 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI ABINGDON PA 4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLAND SN 1042-5179 J9 DNA SEQUENCE JI DNA Seq. PD AUG PY 2002 VL 13 IS 4 BP 211 EP 215 DI 10.1080/1042517021000021590 PG 5 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 595XC UT WOS:000178133000005 PM 12487023 ER PT J AU Shears, SB AF Shears, SB TI Nudix hydrolases active towards diadenosine and diphosphoinositol polyphosphates SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIEHS, Inositol Signaling Sect, NIH, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 18 BP 550 EP 550 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200020 ER PT J AU Sitkovsky, MV AF Sitkovsky, MV TI Engineering inflammation SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 24 BP 551 EP 551 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200026 ER PT J AU Jacobson, KA Liang, BT AF Jacobson, KA Liang, BT TI Engineering of the human A(3) receptor and its agonists for cardioprotection SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIDDK, Mol Recognit Sect, LBC, NIH, Bethesda, MD 20892 USA. Univ Connecticut, Ctr Hlth, Dept Cardiol, Farmington, CT 06030 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 31 BP 552 EP 552 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200033 ER PT J AU Jacobson, KA Kim, HS Xu, B Lee, K Ravi, G Kim, SK Ji, XD Kim, SG Barak, D Marquez, VE Gao, ZG AF Jacobson, KA Kim, HS Xu, B Lee, K Ravi, G Kim, SK Ji, XD Kim, SG Barak, D Marquez, VE Gao, ZG TI Ribose-modified nucleosides and nucleotides as ligands for adenosine and P2 nucleotide receptors SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIDDK, Mol Recognit Sect, LBC, NIH, Bethesda, MD USA. NCI, Med Chem Lab, Frederick, MD 21701 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 0 TC 0 Z9 0 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 45 BP 555 EP 555 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200047 ER PT J AU Sitkovsky, MV AF Sitkovsky, MV TI A(2A) adenosine receptors trigger physiological antiinflammatory pathway. SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 47 BP 555 EP 555 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200049 ER PT J AU Linden, J Figler, RA Jacobson, KA Wang, GQ LaNoue, KF AF Linden, J Figler, RA Jacobson, KA Wang, GQ LaNoue, KF TI Clinical targets of A(2B) adenosine receptor antagonists SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 Univ Virginia, Sch Med, Dept Mol Physiol, Charlottesville, VA 22908 USA. Univ Virginia, Sch Med, Dept Biol Phys, Charlottesville, VA 22908 USA. Univ Virginia, Sch Med, Dept Med, Charlottesville, VA 22908 USA. Adenosine Therapeut LLC, Charlottesville, VA USA. NIH, Bethesda, MD 20892 USA. Penn State Univ, Hershey, PA USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 0 TC 0 Z9 0 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 51 BP 556 EP 556 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200053 ER PT J AU He, M Koshimizu, T Tomic, M Stojilkovic, SS AF He, M Koshimizu, T Tomic, M Stojilkovic, SS TI Ligand binding domain-dependent desensitization of P2X receptors SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NICHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 105 BP 565 EP 565 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200107 ER PT J AU Gaul, ZG Chen, A Barak, D Kim, SK Muller, CF Jacobson, KA AF Gaul, ZG Chen, A Barak, D Kim, SK Muller, CF Jacobson, KA TI Identification by site-directed mutagenesis of residues involved in ligand recognition and activation of human A(3) adenosine receptors SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 NIDDK, Mol Recognit Sect, LBC, NIH, Bethesda, MD 20892 USA. Univ Bonn, Inst Pharmaceut, D-5300 Bonn, Germany. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 0 TC 0 Z9 0 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 107 BP 566 EP 566 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200109 ER PT J AU Wihlberg, AK Malmsjo, M Eyjolfsson, A Gustafsson, R Jacobson, K Erlinge, D AF Wihlberg, AK Malmsjo, M Eyjolfsson, A Gustafsson, R Jacobson, K Erlinge, D TI P2Y(1), P2Y(2/4) and P2Y(6) receptor mediated endothelium dependent vasodilation in human arteries SO DRUG DEVELOPMENT RESEARCH LA English DT Meeting Abstract CT 7th International Symposium on Adenosine and Adenine Nucleotides CY MAY, 2001 CL GOLD COAST, AUSTRALIA SP Abbott Labs, Adenosine Therapeut, Bank Leumi, Biogen, CV Therapeut, Euroscreen, Fjisawa Healthcare Inc, GlaxoSmithKline, Griffith Univ, Inspire Pharmaceut Inc, Johnson & Johnson, King Medco Res Inc, Kyowa Hakko, Merck, Novartis, Pfizer C1 Lund Univ, Dept Cardiol, Ctr Biomed C12, S-22100 Lund, Sweden. NIH, Mol Recognit Sect, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD AUG PY 2002 VL 56 IS 4 MA 129 BP 569 EP 569 PG 1 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 611VJ UT WOS:000179038200131 ER PT J AU Wang, HB Chanas, B Ghanayem, BI AF Wang, HB Chanas, B Ghanayem, BI TI Cytochrome P450 2E1 (CYP2E1) is essential for acrylonitrile metabolism to cyanide: Comparative studies using CYP2E1-null and wild-type mice SO DRUG METABOLISM AND DISPOSITION LA English DT Article ID IN-VIVO; RATS; GLUTATHIONE; TOXICITY; CYSTEINE; MORTALITY; INVITRO; WORKERS; BINDING AB Acrylonitrile (AN) is a rodent carcinogen and suspected human carcinogen. Metabolism of AN proceeds via conjugation with glutathione or epoxidation via cytochrome P4502E1 (CYP2E1) to cyanoethylene oxide (CEO). It was hypothesized that CEO metabolism via epoxide hydrolase (EH) is the primary pathway for cyanide formation. The objective of this work is to assess the enzymatic basis of metabolism to cyanide. Male wild-type and CYP2E1-null mice received 0, 2.5, 10, 20, or 40 mg of AN/kg by gavage, and cyanide was measured in blood and tissues. CYP2E1 and EH expression were assessed using Western blot analyses. Present results demonstrated that cyanide concentrations in blood and tissues of AN-treated wild-type mice were higher at 1 versus 3 h, increased in a dose-dependent manner, and were significantly higher in AN-treated versus vehicle-treated mice. In contrast, cyanide concentrations in the blood and tissues of AN-treated CYP2E1-null mice were not statistically different from those of vehicle-treated mice. Furthermore, this work showed that EH is expressed in CYP2E1-null and wild-type mice. In conclusion, under the current experimental conditions using CYP2E1-null mice, current work demonstrated for the first time that CYP2E1-mediated oxidation is a prerequisite for AN metabolism to cyanide. Since earlier studies showed that CYP2E1 is the only enzyme responsible for AN epoxidation, it is concluded that AN metabolism to CEO is a prerequisite for cyanide formation, and this pathway is exclusively catalyzed by CYP2E1. Finally, this work confirmed that cyanide plays an essential role in the causation of the acute toxicity/mortality of AN. C1 Natl Inst Environm Hlth Sci, Environm Toxicol Program, NIH, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA. RP Ghanayem, BI (reprint author), Natl Inst Environm Hlth Sci, Environm Toxicol Program, NIH, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA. NR 36 TC 39 Z9 44 U1 0 U2 1 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0090-9556 J9 DRUG METAB DISPOS JI Drug Metab. Dispos. PD AUG PY 2002 VL 30 IS 8 BP 911 EP 917 DI 10.1124/dmd.30.8.911 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 574GB UT WOS:000176879400008 PM 12124309 ER PT J AU Xiong, H Yoshinari, K Brouwer, KLR Negishi, M AF Xiong, H Yoshinari, K Brouwer, KLR Negishi, M TI Role of constitutive androstane receptor in the in vivo induction of Mrp3 and CYP2B1/2 by phenobarbital SO DRUG METABOLISM AND DISPOSITION LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Association-of-Pharmaceutical-Scientists CY OCT 21-25, 2001 CL DENVER, COLORADO SP Amer Assoc Pharmaceut Scientists ID RESPONSIVE ENHANCER MODULE; PREGNANE-X-RECEPTOR; FA/FA ZUCKER RAT; ACETAMINOPHEN GLUCURONIDE; 5'-FLANKING REGION; NUCLEAR RECEPTORS; GENE-EXPRESSION; PROTEIN-3 MRP3; CAR; LIVER AB Phenobarbital (PB) induces the hepatic organic anion transporter, Mrp3. The present study tested the hypothesis that Mrp3 induction by PB is mediated by the constitutive androstane receptor (CAR). PB induction of Mrp3 and CYP2B was examined in lean and obese Zucker rats, male and female Wistar Kyoto (WKY) rats, HepG2 and mouse CAR-expressing HepG2 (g2car-3) cells; HepG2 and g2car-3 cells also were treated with 1,4-bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP). In obese Zucker rat livers, total and nuclear CAR levels were markedly lower compared with lean rat livers, which correlated with the poor induction of CYP2B1/2 by PB in obese Zucker rats. Mrp3 induction by PB also was impaired in obese Zucker rat livers. Induction of Mrp3 by PB was similar in male and female WKY rat livers, despite the fact that CAR protein levels were significantly lower in female relative to male WKY rat livers. MRP3 levels in both HepG2 and g2car-3 cells were induced to a similar extent in the two cell lines by PB but not by TCPOBOP. In contrast, CYP2B6 levels were measurable and induced by TCPOBOP only in g2car-3 cells. In conclusion, data from WKY rats and HepG2 cells suggest that CAR does not play a key role in PB induction of Mrp3. Impaired induction of Mrp3 by PB in obese Zucker rats is not due solely to CAR deficiency. Interestingly, differences in the constitutive levels of Mrp3 were observed between obese and lean Zucker rats and between male and female WKY rats. C1 Univ N Carolina, Div Drug Delivery & Disposit, Sch Pharm, Chapel Hill, NC 27599 USA. Natl Inst Environm Hlth Sci, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC USA. RP Brouwer, KLR (reprint author), Univ N Carolina, Div Drug Delivery & Disposit, Sch Pharm, CB 7360,Beard Hall, Chapel Hill, NC 27599 USA. FU NIGMS NIH HHS [R01 GM041935, GM41935] NR 40 TC 83 Z9 83 U1 1 U2 3 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0090-9556 J9 DRUG METAB DISPOS JI Drug Metab. Dispos. PD AUG PY 2002 VL 30 IS 8 BP 918 EP 923 DI 10.1124/dmd.30.8.918 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 574GB UT WOS:000176879400009 PM 12124310 ER PT J AU Larson, VD Williams, DW Henderson, WG Luethke, LE Beck, LB Noffsinger, D Bratt, GW Dobie, RA Fausti, SA Haskell, GB Rappaport, BZ Shanks, JE Wilson, RH AF Larson, VD Williams, DW Henderson, WG Luethke, LE Beck, LB Noffsinger, D Bratt, GW Dobie, RA Fausti, SA Haskell, GB Rappaport, BZ Shanks, JE Wilson, RH TI A multi-center, double blind clinical trial comparing benefit from three commonly used hearing aid circuits SO EAR AND HEARING LA English DT Article AB Objective: Although numerous studies have demonstrated that hearing aids provide significant benefit, carefully controlled, multi-center clinical trials have not been conducted. A multi-center clinical trial was conducted to compare the efficacy of three commonly used hearing aid circuits: peak clipping, compression limiting, and wide dynamic range compression. Conclusions: The three hearing aid circuits studied provide significant benefit both in quiet and in noisy listening situations. The two compression hearing aids appear to provide superior benefits compared to the linear circuit, although the differences between the hearing aids were smaller than the differences between unaided and aided conditions. Design: Patients (N = 360) with bilateral, sensorineural hearing loss were studied using a double blind, three-period, three-treatment crossover design. The patients were fit with each of three programmable hearing aid circuits. Outcome tests were administered in the unaided condition at baseline and then after 3 mo usage of each circuit, the tests were administered in both aided and unaided conditions. The outcome test battery included tests of speech recognition, sound quality and subjective scales of hearing aid benefit, including patients' overall rank-order rating of the three circuits. Results: Each hearing aid circuit improved speech recognition markedly, with greater improvement observed for soft and conversationally loud speech in both quiet and noisy listening conditions. In addition, a significant reduction in the problems encountered in communication was observed. Some tests suggested that the two compression hearing aids provided a better listening experience than the peak clipping hearing aid. In the rank-order ratings, patients preferred the compression limiting hearing aid more frequently than the other two hearing aids. Conclusions: The three hearing aid circuits studied provide significant benefit both in quiet and in noisy listening situations. The two compression hearing aids appear to provide superior benefits compared to the linear circuit, although the differences between the hearing aids were smaller than the differences between unaided and aided conditions. C1 Howard Leight Ind, San Diego, CA 92154 USA. Hines VA Cooperat Studies Program Coordinating Ct, Hines, IL USA. Natl Inst Deafness & Other Commun Dis, Bethesda, MD USA. Greater Los Angeles VA Hlth Care Syst, Los Angeles, CA USA. Univ Texas, Hlth Sci Ctr, San Antonio, TX USA. Dept Vet Affairs Med Ctr, Washington, DC USA. Dept Vet Affairs Med Ctr, Nashville, TN USA. Dept Vet Affairs Med Ctr, Portland, OR USA. Dept Vet Affairs Med Ctr, Iowa City, IA USA. Dept Vet Affairs Med Ctr, Albuquerque, NM USA. Dept Vet Affairs Med Ctr, Long Beach, CA USA. Dept Vet Affairs Med Ctr, Mountain Home, TN USA. RP Larson, VD (reprint author), Howard Leight Ind, 7828 Waterville Rd, San Diego, CA 92154 USA. OI dobie, robert/0000-0003-3833-1772 NR 22 TC 20 Z9 22 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0196-0202 J9 EAR HEARING JI Ear Hear. PD AUG PY 2002 VL 23 IS 4 BP 269 EP 276 DI 10.1097/01.AUD.0000027409.90397.E4 PG 8 WC Audiology & Speech-Language Pathology; Otorhinolaryngology SC Audiology & Speech-Language Pathology; Otorhinolaryngology GA 586CN UT WOS:000177566400001 PM 12195168 ER PT J AU Henderson, WG Larson, VD Williams, D Leuthke, L AF Henderson, WG Larson, VD Williams, D Leuthke, L TI Organization and administration of the NIDCD/VA Hearing Aid Clinical Trial SO EAR AND HEARING LA English DT Article AB This article describes the organization and administration of the NIDCD/VA Hearing Aid Clinical Trial. The trial involved a total of 360 patients with bilateral, sensorineural hearing loss from eight VA Medical Centers to study three different hearing aid circuits in a three-period, three-treatment crossover design. Strong central coordination of such a complex multi-center clinical trial is essential to its success. The trial took more than 5 years to design, implement, and complete. This timeline is also described. C1 Hines VA Cooperat Studies Program Coordinating Ct, Hines, IL USA. Howard Leight Ind, San Diego, CA USA. Natl Inst Deafness & Other Commun Disorder, Bethesda, MD USA. RP Henderson, WG (reprint author), VA Hosp, Cooperat Studies Program, POB 5000-151K, Hines, IL 60141 USA. NR 5 TC 2 Z9 2 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0196-0202 J9 EAR HEARING JI Ear Hear. PD AUG PY 2002 VL 23 IS 4 BP 277 EP 279 DI 10.1097/01.AUD.0000027400.77427.CF PG 3 WC Audiology & Speech-Language Pathology; Otorhinolaryngology SC Audiology & Speech-Language Pathology; Otorhinolaryngology GA 586CN UT WOS:000177566400002 PM 12195169 ER PT J AU Haskell, GB Noffsinger, D Larson, VD Williams, DW Dobie, RA Rogers, JL AF Haskell, GB Noffsinger, D Larson, VD Williams, DW Dobie, RA Rogers, JL TI Subjective measures of hearing aid benefit in the NIDCD/VA Clinical Trial SO EAR AND HEARING LA English DT Article AB Objective. Subjective measures of performance were assessed on three different hearing aid circuits as part of a large clinical trial. These measurements included the Profile of Hearing Aid Performance and a subjective ranking of individual preference. Design: A multi-center, double-masked clinical trial of hearing aids was conducted at eight VA Medical Centers. Three hearing aid circuits, a linear peak-clipper, a linear compression limiter and a wide dynamic range compressor, were investigated. The experimental design was a three-period, three-treatment crossover design. Subjects (N = 360) were stratified by site and randomized to one of six sequences for the hearing aid circuits. All fittings were binaural and involved a 3-mo trial with each of the three circuits. All subjective measures were administered for unaided and aided conditions at the end of each trial period. Results: While all of the circuits resulted in improved scores on the aided versus the unaided PHAP, there were few conditions in which one circuit outperformed the others. An exception was the aversiveness of sound subscale where the peak clipper frequently scored worse than either the compression limiter or the wide dynamic range compressor. In the subjective ranking scale the compression limiter received more first place rankings than the other two circuits, especially for one subgroup of patients with moderate flat hearing loss. Conclusions: All circuits were perceived as beneficial by these subjects in most situations. The peak clipper scored worse on aversiveness of sound than did the other two circuits for most subjects, while the compression limiter seemed to have a slight advantage in subjective rankings. Most subjects perceived considerable aided benefit in situations involving background noise and reverberation, situations where hearing aid benefit is often questioned. C1 VA Med Ctr, Iowa City, IA USA. VA Med Ctr, Washington, DC USA. VA Greater Los Angeles Hlth Care Syst, Los Angeles, CA USA. Howard Leight Ind, San Diego, CA USA. VA Hines Cooperat Studies Program Coordinating Ct, Hines, IL USA. NIDCD, NIH, Bethesda, MD USA. RP Haskell, GB (reprint author), 8501 E Alameda Ave,Apt 1328, Denver, CO 80230 USA. OI dobie, robert/0000-0003-3833-1772 NR 5 TC 9 Z9 14 U1 2 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0196-0202 J9 EAR HEARING JI Ear Hear. PD AUG PY 2002 VL 23 IS 4 BP 301 EP 307 DI 10.1097/01.AUD.0000027404.26058.47 PG 7 WC Audiology & Speech-Language Pathology; Otorhinolaryngology SC Audiology & Speech-Language Pathology; Otorhinolaryngology GA 586CN UT WOS:000177566400005 PM 12195172 ER PT J AU Wilson, HR Yu, DG Peters, HK Zhou, JG Court, DL AF Wilson, HR Yu, DG Peters, HK Zhou, JG Court, DL TI The global regulator RNase III modulates translation repression by the transcription elongation factor N SO EMBO JOURNAL LA English DT Article DE antitermination; bacteriophage lambda; growth rate regulation; RNase III; translation repression ID ESCHERICHIA-COLI; BACTERIOPHAGE-LAMBDA; COLIPHAGE-LAMBDA; MESSENGER-RNA; GENE-EXPRESSION; CIII-GENE; ANTI-TERMINATION; LARGE PRECURSOR; RIBOSOMAL-RNA; PHAGE-LAMBDA AB Efficient expression of most bacteriophage lambda early genes depends upon the formation of an antiterminating transcription complex to overcome transcription terminators in the early operons, p(L) and p(R). Formation of this complex requires the phage-encoded protein N, the first gene product expressed from the p(L) operon. The N leader RNA contains, in this order: the NUTL site, an RNase III-sensitive hairpin and the N ribosome-binding site. N bound to NUTL RNA is part of both the antitermination complex and an autoregulatory complex that represses the translation of the N gene. In this study, we show that cleavage of the N leader by RNase III does not inhibit antitermination but prevents N-mediated translation repression of N gene expression. In fact, by preventing N autoregulation, RNase III activates N gene translation at least 200-fold. N-mediated translation repression is extremely sensitive to growth rate, reflecting the growth rate regulation of RNase III expression itself. Given N protein's critical role in lambda development, the level of RNase III activity therefore serves as an important sensor of physiological conditions for the bacteriophage. C1 NCI Frederick, Mol Control & Genet Sect, Frederick, MD 21702 USA. RP Court, DL (reprint author), NCI Frederick, Mol Control & Genet Sect, Frederick, MD 21702 USA. NR 42 TC 28 Z9 30 U1 1 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD AUG 1 PY 2002 VL 21 IS 15 BP 4154 EP 4161 DI 10.1093/emboj/cdf395 PG 8 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 584MQ UT WOS:000177470700023 PM 12145215 ER PT J AU Jones, JM Gellert, M AF Jones, JM Gellert, M TI Ordered assembly of the V(D)J synaptic complex ensures accurate recombination SO EMBO JOURNAL LA English DT Article DE DNA recombination; nucleoprotein complex; RAG1; RAG2; V(D)J recombination ID LAMBDA INTEGRATIVE RECOMBINATION; BROKEN DNA-MOLECULES; DOUBLE-STRAND BREAKS; SINGLE ACTIVE-SITE; TARGETED DELETION; FUNCTIONAL-ANALYSIS; CLEAVAGE COMPLEXES; SIGNAL SEQUENCES; MOUSE THYMOCYTES; NUCLEOSOMAL DNA AB Recombination of gene segments at the immunoglobulin and T-cell receptor loci requires that the RAG1 and RAG2 proteins bring together DNA signal sequences (RSSs) with 12- and 23-bp spacers into a synaptic complex and cleave the DNA. A RAG1/2 multimer that can cleave both signals is shown to assemble on an isolated RSS, and the complementary RSS enters this complex as naked DNA. When RAG1/ 2 is allowed to bind 12 and 23 RSSs separately prior to their mixing, synaptic complex assembly and cleavage activity are greatly reduced, indicating that only a complex initially assembled on a single RSS leads to productive cleavage. RAG1/2 complexes assembled on 12 RSSs will only incorporate, 23 partners, while complexes assembled on 23 RSSs show a 5- to 6-fold preference for 12 partners. Thus, initial assembly on a 12 RSS most accurately reflects the strict 12/23 coupled cleavage observed in the cell. Additional cellular factors such as chromatin may ensure that RAG1/2 first assembles on a 12 RSS, and then a free 23 RSS enters to activate cleavage. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Gellert, M (reprint author), NIDDKD, Mol Biol Lab, NIH, Bldg 5,Room 241, Bethesda, MD 20892 USA. NR 50 TC 75 Z9 77 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD AUG 1 PY 2002 VL 21 IS 15 BP 4162 EP 4171 DI 10.1093/emboj/cdf394 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 584MQ UT WOS:000177470700024 PM 12145216 ER PT J AU Catez, F Brown, DT Misteli, T Bustin, M AF Catez, F Brown, DT Misteli, T Bustin, M TI Competition between histone H1 and HMGN proteins for chromatin binding site SO EMBO REPORTS LA English DT Article ID CHICKEN ERYTHROCYTE CHROMATIN; CHROMOSOMAL-PROTEINS; HIGH-MOBILITY; HMG-14; NUCLEOSOMES; ACTIVATION; NUCLEUS; CELLS; H-1 AB The ability of regulatory factors to access their nucleosomal targets is modulated by nuclear proteins such as histone Fill and HMGN (previously named HMG-14/-17 family) that bind to nucleosomes and either stabilize or destabilize the higher-order chromatin structure. We tested whether HMGN proteins affect the interaction of histone H1 with chromatin. Using microinjection into living cells expressing H1-GFP and photobleaching techniques, we found that wild-type HMGN, but not HMGN point mutants that do not bind to nucleosomes, inhibits the binding of H1 to nucleosomes. HMGN proteins compete with H1 for nucleosome sites but do not displace statically bound H1 from chromatin. Our results provide evidence for in vivo competition among chromosomal proteins for binding sites on chromatin and suggest that the local structure of the chromatin fiber is modulated by a dynamic interplay between nucleosomal binding proteins. C1 NCI, NIH, Bethesda, MD 20892 USA. Univ Mississippi, Med Ctr, Jackson, MS 39216 USA. RP Bustin, M (reprint author), NCI, NIH, Bethesda, MD 20892 USA. RI Bustin, Michael/G-6155-2015 NR 26 TC 96 Z9 101 U1 0 U2 2 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1469-221X J9 EMBO REP JI EMBO Rep. PD AUG PY 2002 VL 3 IS 8 BP 760 EP 766 DI 10.1093/embo-reports/kvf156 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 586LK UT WOS:000177585300016 PM 12151335 ER PT J AU Sun, YH Gershengorn, MC AF Sun, YH Gershengorn, MC TI Correlation between basal signaling and internalization of thyrotropin-releasing hormone receptors: Evidence for involvement of similar receptor conformations SO ENDOCRINOLOGY LA English DT Article ID AGONIST-INDUCED INTERNALIZATION; BETA-ADRENERGIC-RECEPTOR; C-TERMINAL TAIL; HUMAN LUTROPIN/CHORIOGONADOTROPIN RECEPTOR; PROTEIN-COUPLED RECEPTORS; BETA(2)-ADRENERGIC RECEPTOR; INTRACELLULAR LOOP; MOLECULAR-CLONING; PHOSPHOLIPASE-C; STIMULATED INTERNALIZATION AB Previous studies have shown that rat thyrotropin-releasing hormone (TRH) receptor type 2 exhibits higher basal signaling activity and internalizes more rapidly upon agonist binding than rat TRH receptor type 1. The mouse TRH receptor type 2 (mR2) was recently cloned and, similar to its rat homolog, shows a higher basal signaling activity than mR1. Taking advantage of the high degree of sequence homology between mR1 and mR2, we used chimeras/mutants of these receptors to gain insight into the properties of the receptors that influence internalization and basal signaling. Chimeric receptors that have the mR1 extracellular and transmembrane domains with the carboxyl terminus and intracellular loops of mR2 (R1/R2-tail; R1/R2-I3,tail; R1/R2-I2,3,tail; R1/R2-I1,2,3,tail) exhibited internalization rates and basal activities that were similar to that of mR1. In contrast, a chimeric receptor with the extracellular and transmembrane domains of mR2 and the carboxyl terminus of mR1 exhibited the more rapid internalization rate and higher basal signaling activity characteristic of mR2. We showed previously that mutation of a highly conserved tryptophan to alanine caused mR1 to exhibit a high basal signaling activity and rapid internalization rate. In contrast, mutation of this tryptophan to alanine in mR2 decreased the rate of internalization and inhibited basal signaling activity. The rates of receptor internalization did not correlate with the binding affinities, coupling efficiencies, or potencies of the receptors. Thus, we observed that receptors with more rapid internalization rates showed relatively higher basal signaling activities, whereas receptors with lower basal signaling activities showed slower internalization rates. These data suggest that similar receptor conformations are required for productive coupling to signaling G proteins and to proteins involved in internalization. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. Cornell Univ, Weill Med Coll, Dept Med, Div Mol Med, New York, NY 10021 USA. RP Gershengorn, MC (reprint author), NIDDKD, Diabet Branch, NIH, 50 South Dr,MSC 8029, Bethesda, MD 20892 USA. FU NIDDK NIH HHS [DK 43031] NR 63 TC 12 Z9 13 U1 0 U2 0 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD AUG PY 2002 VL 143 IS 8 BP 2886 EP 2892 DI 10.1210/en.143.8.2886 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 576JC UT WOS:000176999600008 PM 12130552 ER PT J AU Watson, PA Vinson, C Nesterova, A Reusch, JEB AF Watson, PA Vinson, C Nesterova, A Reusch, JEB TI Content and activity of cAMP response element-binding protein regulate platelet-derived growth factor receptor-alpha content in vascular smooth muscles SO ENDOCRINOLOGY LA English DT Article ID CELL-PROLIFERATION; TRANSCRIPTIONAL REGULATION; GENE-EXPRESSION; PPAR-GAMMA; MIGRATION; PDGF; DELTA; ATHEROSCLEROSIS; TROGLITAZONE; RATS AB Experiments in vascular smooth muscle cells (SMCs) indicate that the transcription factor cAMP response element-binding protein (CREB), the cyclic nucleotide response element-binding protein, suppresses expression of the platelet-derived growth factor-a receptor gene (PDGFRalpha). Adenovirus-mediated expression of constitutively active CREB mutants decreases PDGFRalpha mRNA, PDGFRalpha protein, and PDGFRalpha promoter-luciferase reporter activity in cultured SMCs. Expression of dominant negative CREB protein, A-CREB, increases PDGFRalpha protein content and the PDGFRalpha-promoter activity in SMCs. Active CREB prevents activation of PDGFRalpha promoter-luciferase reporter activity by CCAAT/enhancer-binding protein-delta (C/EBPdelta), shown to mediate IL-1beta stimulation of PDGFRalpha expression. Exposure of cultured SMCs to high glucose or reactive oxidant stress, which decrease CREB protein content and activity, increases PDGFRalpha protein content and promoter activity. Expression of active CREB blunts reactive oxidant stress-induced PDGFRalpha accumulation in SMCs. Loss of CREB protein in aortic walls of rats with streptozotocin-induced diabetes is accompanied by an increase in PDGFRalpha content. In Ob/Ob mice (which demonstrate reduced aortic wall CREB content vs. Ob/- controls), treatment with the peroxisomal proliferator-activated receptor gamma rosiglitazone increases CREB content and decreases PDGFRalpha content in the aortic wall. Thus, both in vitro and in vivo loss of CREB content and activity and subsequent accumulation of PDGFRalpha may contribute to SMC activation during diabetes. C1 Denver VA Med Ctr, Denver Res Inst, Denver, CO 80220 USA. Univ Colorado, Hlth Sci Ctr, Div Endocrinol, Denver, CO 80220 USA. Univ Colorado, Hlth Sci Ctr, Dept Med, Denver, CO 80220 USA. NCI, NIH, Bethesda, MD 20892 USA. RP Reusch, JEB (reprint author), Denver VA Med Ctr, Denver Res Inst, Room 9C-120,1055 Clermont St, Denver, CO 80220 USA. NR 22 TC 14 Z9 14 U1 0 U2 0 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD AUG PY 2002 VL 143 IS 8 BP 2922 EP 2929 DI 10.1210/en.143.8.2922 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 576JC UT WOS:000176999600013 PM 12130557 ER PT J AU Cheng, RYS Alvord, WG Powell, D Kasprzak, KS Anderson, LM AF Cheng, RYS Alvord, WG Powell, D Kasprzak, KS Anderson, LM TI Increased serum corticosterone and glucose in offspring of chromium(III)-treated male mice SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE chromium(III); corticosterone; insulin-like growth factor 1; insulin-like growth factor binding protein 1; offspring; paternal; preconceptional; stress; transgenerational ID FACTOR BINDING-PROTEINS; GROWTH-FACTOR; CALORIC RESTRICTION; EPIDEMIOLOGIC EVIDENCE; BREAST-CANCER; INSULIN; RISK; DIRECTIONS; ADENOMAS; TUMORS AB Preconceptional carcinogenesis occurs in animals and is suspected for humans-for example, after occupational metals exposure. Several characteristics in animal models, including high frequency and non-Mendelian inheritance patterns, have suggested an epigenetic mechanism, possibly involving hormone changes in offspring. To test this hypothesis, we treated male mice with chromium(III) chloride, a preconceptional carcinogen, 2 weeks before mating, in two separate experiments. Their 10-week-old offspring showed highly significant increases in average serum corticosterone and glucose, compared with control offspring. Average serum levels of insulin-like growth factor 1 (IGF1) showed more modest possible increases. A previous microarray experiment identified hepatic insulin-like growth factor binding protein 1 (IGF BP1) gene expression as consistently changed in correlation with serum corticosterone levels. In the present study, hepatic IGF BP1 mRNA correlated with serum IGF1 in male offspring of chromium-treated fathers, but not in controls; serum glucose correlated positively with hepatic IGF BP1 in chromium-group offspring but negatively in controls. These results support the hypothesis that preconceptional exposure effects may alter hormones, metabolism, and control of tissue gene expression, probably through epigenetic mechanisms. Risk of neoplasia may be influenced by these changes. C1 NCI, Comparat Carcinogenesis Lab, Ctr Canc Res, Frederick, MD 21702 USA. NCI, Data Management Serv Inc, Frederick, MD 21702 USA. RP Anderson, LM (reprint author), NCI, Comparat Carcinogenesis Lab, Ctr Canc Res, Bldg 538, Frederick, MD 21702 USA. OI Cheng, Robert/0000-0003-0287-6439 NR 35 TC 16 Z9 17 U1 0 U2 1 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD AUG PY 2002 VL 110 IS 8 BP 801 EP 804 PG 4 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 583FK UT WOS:000177395900035 PM 12153762 ER PT J AU Crook, MF Boehm, M Yoshimoto, T Akyurek, LM Olive, M Ganesh, S Nabel, EG AF Crook, MF Boehm, M Yoshimoto, T Akyurek, LM Olive, M Ganesh, S Nabel, EG TI Molecular cloning and characterization of the human KIS gene promoter - a regulator of vascular smooth muscle cell proliferation SO EUROPEAN HEART JOURNAL LA English DT Meeting Abstract CT Congress of the European-Society-of-Cardiology CY AUG 31-SEP 04, 2002 CL BERLIN, GERMANY SP European Soc Cardiol C1 NIH, Vasc Biol Branch, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU W B SAUNDERS CO LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0195-668X J9 EUR HEART J JI Eur. Heart J. PD AUG-SEP PY 2002 VL 23 SU S BP 133 EP 133 PG 1 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 624HB UT WOS:000179753300499 ER PT J AU Neuss, M Chesley, AT Crow, MT Fleck, E AF Neuss, M Chesley, AT Crow, MT Fleck, E TI The apoptosis regulator ARC prevents apoptotic and necrotic cell death in cardiomyocytes SO EUROPEAN HEART JOURNAL LA English DT Meeting Abstract CT Congress of the European-Society-of-Cardiology CY AUG 31-SEP 04, 2002 CL BERLIN, GERMANY SP European Soc Cardiol C1 German Heart Inst, Dept Cardiol, Berlin, Germany. NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU W B SAUNDERS CO LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0195-668X J9 EUR HEART J JI Eur. Heart J. PD AUG-SEP PY 2002 VL 23 SU S BP 245 EP 245 PG 1 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 624HB UT WOS:000179753300923 ER PT J AU Feher, A Weber, IT Bagossi, P Boross, P Mahalingam, B Louis, JM Copeland, TD Torshin, IY Harrison, RW Tozser, J AF Feher, A Weber, IT Bagossi, P Boross, P Mahalingam, B Louis, JM Copeland, TD Torshin, IY Harrison, RW Tozser, J TI Effect of sequence polymorphism and drug resistance on two HIV-1 Gag processing sites SO EUROPEAN JOURNAL OF BIOCHEMISTRY LA English DT Article DE HIV-1 proteinase; Gag processing sites; oligopeptide substrates; substrate specificity; molecular modeling ID IMMUNODEFICIENCY-VIRUS TYPE-1; PROTEASE CLEAVAGE SITE; RETROVIRAL PROTEINASES; CROSS-RESISTANCE; VIRAL FITNESS; IN-VIVO; INHIBITORS; MUTATIONS; MUTANTS; INDINAVIR AB The HIV-1 proteinase (PR) has proved to be a good target for antiretroviral therapy of AIDS, and various PR inhibitors are now in clinical use. However, there is a rapid selection of viral variants bearing mutations in the proteinase that are resistant to clinical inhibitors. Drug resistance also involves mutations of the nucleocapsid/p1 and p1/p6 cleavage sites of Gag, both in vitro and in vivo. Cleavages at these sites have been shown to be rate limiting steps for polyprotein processing and viral maturation. Furthermore, these sites show significant sequence polymorphism, which also may have an impact on virion infectivity. We have studied the hydrolysis of oligopeptides representing these cleavage sites with representative mutations found as natural variations or that arise as resistant mutations. Wild-type and five drug resistant PRs with mutations within or outside the substrate binding site were tested. While the natural variations showed either increased or decreased susceptibility of peptides toward the proteinases, the resistant mutations always had a beneficial effect on catalytic efficiency. Comparison of the specificity changes obtained for the various substrates suggested that the maximization of the van der Waals contacts between substrate and PR is the major determinant of specificity: the same effect is crucial for inhibitor potency. The natural nuclcocapsid/p1 and p1/p6 sites do not appear to be optimized for rapid hydrolysis. Hence, mutation of these rate limiting cleavage sites can partly compensate for the reduced catalytic activity of drug resistant mutant HIV-1 proteinases. C1 Univ Debrecen, Fac Med, Dept Biochem & Mol Biol, H-4012 Debrecen, Hungary. Georgia State Univ, Dept Biol, Atlanta, GA USA. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. NCI, Frederick, MD 21701 USA. Georgia State Univ, Dept Comp Sci, Atlanta, GA 30303 USA. RP Tozser, J (reprint author), Univ Debrecen, Fac Med, Dept Biochem & Mol Biol, POB 6, H-4012 Debrecen, Hungary. EM tozser@indi.biochem.dote.hu RI Tozser, Jozsef/A-7840-2008; OI Tozser, Jozsef/0000-0003-0274-0056; Tozser, Jozsef/0000-0001-5076-8729 FU FIC NIH HHS [TW01001]; NIGMS NIH HHS [GM62920, R01 GM062920] NR 50 TC 57 Z9 58 U1 0 U2 1 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0014-2956 J9 EUR J BIOCHEM JI Eur. J. Biochem. PD AUG PY 2002 VL 269 IS 16 BP 4114 EP 4120 DI 10.1046/j.1432-1033.2002.03105.x PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 589VT UT WOS:000177782300026 PM 12180988 ER PT J AU Zheng, T Holford, TR Mayne, ST Owens, PH Boyle, P Zhang, B Zhang, YW Zahm, SH AF Zheng, T Holford, TR Mayne, ST Owens, PH Boyle, P Zhang, B Zhang, YW Zahm, SH TI Use of hair colouring products and breast cancer risk: a case-control study in Connecticut SO EUROPEAN JOURNAL OF CANCER LA English DT Article DE breast cancer; case-control study; epidemiology; hair dye products ID DYE USE; COLORING PRODUCTS; MULTIPLE-MYELOMA; EXPOSURE; LYMPHOMA; WOMEN AB This case-control study was designed to test the hypothesis that the risk of breast cancer varies by type and colour of the hair colouring products used. A total of 608 cases and 609 controls were included in the study. We found no increased risk associated with the overall use of hair dye products or exclusive use of permanent or temporary types of hair dye products. Among those who reported to have exclusively used semi-permanent types of hair colouring products, some of the ORs were elevated. However, none of the ORs related to age at first use, duration of use, total number of applications, and-years since first use, was statistically significant. There was also no increased risk of breast cancer associated with exclusive use of dark or light hair colouring products, or use of mixed types or colours of hair dye products. We also found no increased risk of breast cancer associated with hair dye use based on an individual's reason for using a hair colouring product, such as to cover grey-or to change natural hair colour. These data suggest that the use of hair colouring products does not have a major impact on the risk of breast cancer. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Yale Univ, Sch Publ Hlth, New Haven, CT 06520 USA. European Inst Oncol, Dept Epidemiol & Biostat, I-20141 Milan, Italy. McGill Univ, Dept Epidemiol & Biostat, Montreal, PQ H3A 1A2, Canada. NCI, Div Epidemiol & Genet Sci, Bethesda, MD 20892 USA. RP Zheng, T (reprint author), Yale Univ, Sch Publ Hlth, 60 Coll St, New Haven, CT 06520 USA. RI Boyle, Peter/A-4380-2014; Zahm, Shelia/B-5025-2015 OI Boyle, Peter/0000-0001-6251-0610; FU NCI NIH HHS [CA-62986] NR 28 TC 16 Z9 17 U1 0 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0959-8049 J9 EUR J CANCER JI Eur. J. Cancer PD AUG PY 2002 VL 38 IS 12 BP 1647 EP 1652 AR PII S0959-8049(02)00138-7 DI 10.1016/S0959-8049(02)00138-7 PG 6 WC Oncology SC Oncology GA 584ZQ UT WOS:000177499000025 PM 12142056 ER PT J AU Ashok, BT Chen, YG Liu, X Garikapaty, VPS Seplowitz, R Tschorn, J Roy, K Mittelman, A Tiwari, RK AF Ashok, BT Chen, YG Liu, X Garikapaty, VPS Seplowitz, R Tschorn, J Roy, K Mittelman, A Tiwari, RK TI Multiple molecular targets of indole-3-carbinol, a chemopreventive anti-estrogen in breast cancer SO EUROPEAN JOURNAL OF CANCER PREVENTION LA English DT Article; Proceedings Paper CT International Symposium on Antimutagenesis and Anticarcinogenesis CY APR 25-26, 2002 CL NY MED COLL, VALHALLA, NEW YORK SP Eastman Kodak Co, Unilever Best Foods, Pepsi Cola Co, Proctor & Gamble HO NY MED COLL DE protein-DNA interaction; natural anti-estrogens; gel shift assay; indole-3-carbinol; chemopreventive ID ACTIVATED PROTEIN-KINASE; DIETARY INDOLE-3-CARBINOL; RECEPTOR; CELLS; PHOSPHORYLATION; ANTIESTROGENS; EXPRESSION; MECHANISMS; PATHWAY; CARCINOGENESIS AB The mechanism of action of the anti-estrogen indole-3-carbinol (I3C), present in cruciferous vegetables, is being examined in our laboratory with a view to promote the use of this naturally occurring chemopreventive as an alternative to synthetic anti-estrogens in human breast cancer. Our previous results clearly demonstrated that despite its low affinity for the estrogen receptor (ER), I3C abrogated estradiol-mediated cellular and biochemical effects in estradiol-responsive cells and tissues. In an earlier report, we identified ER phosphorylation as one of the targets of I3C, and in this communication we describe the consequence of inhibition of ER phosphorylation. Estradiol-induced DNA-binding proteins that bound to several DNA-responsive elements were inhibited by I3C and this effect was not at the level of DNA-protein physical interaction as inclusion of I3C in vitro in the reaction mix did not affect the binding. We analyzed the spectrum of genes induced by estradiol and modulated and/or intercepted by I3C. Our results conclude that although estradiol-mediated functions are affected by I3C, its biochemical targets are multiple and some of these may be modulated by the oligomeric products of I3C. (C) 2002 Lippincott Williams Wilkins. C1 New York Med Coll, Dept Microbiol & Immunol, Valhalla, NY 10595 USA. New York Med Coll, Dept Microbiol Immunol & Med, Valhalla, NY 10595 USA. NCI, Dev Therapeut Program, Div Canc Treatment & Diag, NIH, Bethesda, MD 20892 USA. RP Tiwari, RK (reprint author), New York Med Coll, Dept Microbiol & Immunol, Valhalla, NY 10595 USA. NR 32 TC 28 Z9 28 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-8278 J9 EUR J CANCER PREV JI Eur. J. Cancer Prev. PD AUG PY 2002 VL 11 SU 2 BP S86 EP S93 PG 8 WC Oncology SC Oncology GA 614QC UT WOS:000179200000013 PM 12570340 ER PT J AU Greenwald, P McDonald, SS Anderson, DE AF Greenwald, P McDonald, SS Anderson, DE TI An evidence-based approach to cancer prevention clinical trials SO EUROPEAN JOURNAL OF CANCER PREVENTION LA English DT Article; Proceedings Paper CT International Symposium on Antimutagenesis and Anticarcinogenesis CY APR 25-26, 2002 CL NY MED COLL, VALHALLA, NEW YORK SP Eastman Kodak Co, Unilever Best Foods, Pepsi Cola Co, Proctor & Gamble HO NY MED COLL DE carcinogenesis; chemoprevention; clinical trials ID COOKED FOODS; INHIBITION; MUTAGENS; DIET; RATS; BEEF; FAT; 1,2-DIMETHYLHYDRAZINE; CARCINOGENESIS; RETINOIDS AB Research on carcinogenesis and its inhibition has made significant progress in the last 30 years, providing an impressive body of evidence that supports various strategies for cancer prevention. Innovative studies have helped to identify potential causes of cancer, including environmental factors such as diet, and provided valuable information about their mechanisms of action. Hundreds of epidemiologic and experimental studies have focused on possible associations between dietary factors and different types of cancer. During the same period, potential inhibitors of cancer that appeared able to prevent, arrest or reverse cancer development by interfering with one or more steps in the process of carcinogenesis were identified, and the term 'chemoprevention' was coined for this pharmacological approach to cancer prevention. Promising compounds were systematically evaluated for their potential as chemopreventive agents. Numerous agents were determined to be safe and effective in preclinical trials, including naturally occurring vitamins, minerals and phytochemicals as well as synthetic compounds. Based on preclinical results, selected agents have been and are now being evaluated in phase I, II and III clinical interventions for various cancers. Development of valid surrogate end point biomarkers for clinical disease that can be modulated by interventions is essential to accelerate progress in cancer prevention clinical trials. (C) 2002 Lippincott Williams Wilkins. C1 NCI, Div Canc Prevent, Bethesda, MD 20892 USA. Sci Consulting Grp Inc, Gaithersburg, MD 20878 USA. RP Greenwald, P (reprint author), NCI, Div Canc Prevent, 6130 Execut Blvd,Suite 2040, Bethesda, MD 20892 USA. NR 25 TC 3 Z9 3 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-8278 J9 EUR J CANCER PREV JI Eur. J. Cancer Prev. PD AUG PY 2002 VL 11 SU 2 BP S43 EP S47 PG 5 WC Oncology SC Oncology GA 614QC UT WOS:000179200000007 PM 12570334 ER PT J AU BenMohamed, L Belkaid, Y Loing, E Brahimi, K Gras-Masse, H Druilhe, P AF BenMohamed, L Belkaid, Y Loing, E Brahimi, K Gras-Masse, H Druilhe, P TI Systemic immune responses induced by mucosal administration of lipopeptides without adjuvant SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE lipopeptide; mucosal immunization; systemic B and T cell responses; dendritic cell; macrophage ID LIVER STAGE ANTIGEN; CHOLERA-TOXIN; INTRANASAL IMMUNIZATION; ANTIBODY-RESPONSES; DENDRITIC CELLS; MINIMAL EPITOPE; T-HELPER; MICE; INDUCTION; IMMUNOGENICITY AB We recently reported that parenteral injection of malaria palmitoyl-tailed peptides without adjuvant efficiently induces B, Th and CTL responses. We now show that intranasal (IN) or sub-lingual (SL) delivery of such lipopeptides induces strong systemic immune responses, as demonstrated by specific Th cell responses from the spleen as well as inguinal lymph nodes, and by the production of high levels of serum antibodies. Overall, both types of responses were significantly higher than in parallel experiments in which the same lipopeptides were delivered by the subcutaneous (s.c.) route. Moreover, the mucosal route resulted in the preferential induction of IFN-gamma producing T cells and of IgG2a antibody production, as compared to the dominant IL-4 and IgG1 responses obtained by the s.c. route, thus bringing a distinct advantage in the field of many infectious diseases and allergy. Possibly related to this Th1 response, we found that dendritic cells, the principal immune-competent cells to encounter antigens within mucosal membranes, take up lipopeptide antigens more efficiently than macrophages. Mucosal immunization by lipidated peptides appears therefore as a novel, noninvasive vaccine approach that does not require the use of extraneous adjuvant and which, besides cost-effectiveness, has attractive practical and immunological features. C1 Inst Pasteur, Biomed Parasitol Unit, F-75724 Paris 15, France. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. Inst Biol, UMR 8525, Lille, France. Inst Pasteur, F-59019 Lille, France. RP Druilhe, P (reprint author), Inst Pasteur, Biomed Parasitol Unit, 28 Rue Dr Roux, F-75724 Paris 15, France. NR 24 TC 73 Z9 76 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI WEINHEIM PA PO BOX 10 11 61, D-69451 WEINHEIM, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD AUG PY 2002 VL 32 IS 8 BP 2274 EP 2281 DI 10.1002/1521-4141(200208)32:8<2274::AID-IMMU2274>3.0.CO;2-C PG 8 WC Immunology SC Immunology GA 586VE UT WOS:000177605900020 PM 12209640 ER PT J AU Rubio-Godoy, V Ayyoub, M Dutoit, V Servis, C Schink, A Rimoldi, D Romero, P Cerottini, JC Simon, R Zhao, YD Houghten, RA Pinilla, C Valmori, D AF Rubio-Godoy, V Ayyoub, M Dutoit, V Servis, C Schink, A Rimoldi, D Romero, P Cerottini, JC Simon, R Zhao, YD Houghten, RA Pinilla, C Valmori, D TI Combinatorial peptide library-based identification of peptide ligands for tumor-reactive cytolytic T lymphocytes of unknown specificity SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE peptide library; tumor immunity; CTL; SSX-2; antigen/peptide/epitope ID ANTIGENIC PEPTIDE; MELANOMA; EXPRESSION; TESTIS; CANCER; GENES; SSX; TCR AB A novel approach for the identification of tumor antigen-derived sequences recognized by CD8(+) cytolytic T lymphocytes (CTL) consists in using synthetic combinatorial peptide libraries. Here we have screened a library composed of 3.1 x 10(11) nonapeptides arranged in a positional scanning format, in a cytotoxicity assay, to search the antigen recognized by melanoma-reactive CTL of unknown specificity. The results of this analysis enabled the identification of several optimal peptide ligands, as most of the individual nonapeptides deduced from the primary screening were efficiently recognized by the CTL. The results of the library screening were also analyzed with a mathematical approach based on a model of independent and additive contribution of individual amino acids to antigen recognition. This biometrical data analysis enabled the retrieval, in public databases, of the native antigenic peptide SSX-2(41-49), whose sequence is highly homologous to the ones deduced from the library screening, among the ones with the highest stimulatory score. These results underline the high predictive value of positional scanning synthetic combinatorial peptide library analysis and encourage its use for the identification of CTL ligands. C1 Torrey Pines Inst Mol Studies, San Diego, CA USA. NCI, Mol Stat & Bioinformat Sect, Biometr Res Branch, NIH, Bethesda, MD 20892 USA. Univ Lausanne, Lausanne Branch, Ludwig Inst Canc Res, CH-1066 Epalinges, Switzerland. Mixture Sci Inc, San Diego, CA USA. Univ Lausanne, Inst Biochem, CH-1066 Epalinges, Switzerland. CHU Vaudois, Ludwig Inst Canc Res, Div Clin Oncol, Univ Hosp, CH-1011 Lausanne, Switzerland. RP Valmori, D (reprint author), Hop Orthopedie, Div Clin Oncoimmunol, Ave Pierre Decker 4, CH-1005 Lausanne, Switzerland. RI Ayyoub, Maha/G-2778-2013; Valmori, Danila/K-2439-2015; Ayyoub, Maha/A-2074-2017 NR 18 TC 26 Z9 28 U1 1 U2 2 PU WILEY-V C H VERLAG GMBH PI WEINHEIM PA PO BOX 10 11 61, D-69451 WEINHEIM, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD AUG PY 2002 VL 32 IS 8 BP 2292 EP 2299 DI 10.1002/1521-4141(200208)32:8<2292::AID-IMMU2292>3.0.CO;2-K PG 8 WC Immunology SC Immunology GA 586VE UT WOS:000177605900022 PM 12209642 ER PT J AU Eddington, ND Cox, DS Roberts, RR Butcher, RJ Edafiogho, IO Stables, JP Cooke, N Goodwin, AM Smith, CA Scott, KR AF Eddington, ND Cox, DS Roberts, RR Butcher, RJ Edafiogho, IO Stables, JP Cooke, N Goodwin, AM Smith, CA Scott, KR TI Synthesis and anticonvulsant activity of enaminones. 4. Investigations on isoxazole derivatives SO EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article DE enaminones; isoxazoles; maximal electroshock seizure test; anticonvulsant activity; X-ray crystallography; structure-activity relationship; physicochemical permeability ID BLOOD-BRAIN-BARRIER; ANTIEPILEPTIC DRUG DEVELOPMENT; CENTRAL-NERVOUS-SYSTEM; TRANSPORT; BINDING; PERMEABILITY; SEIZURES; AGENTS AB Due to the exceptional anticonvulsant activity displayed by substituted aniline enaminones, related pyridine derivatives and phenothiazines synthesised in our laboratories, the further investigation of various aromatic heterocycles was undertaken. Condensation of cyclic 1,3-diketo esters with 3-, and 5-aminoisoxazole derivatives led to,a series of potent anti-maximal electroshock (MES) analogues, three of which occurred in the 3-amino series: ethyl ester (10), orally (po) active in rats [ED(50) 68.9 mg kg(-1), TD(50) > 500 mg kg(-1), protective index (PI = TD(50)/ED(50)) > 49.6]; methyl ester (9), ED(50) 68.9 mg kg(-1) intraperitoneally (ip) in mice, TD(50) > 500 mg kg(-1), PI > 7.3, and tert-butyl ester (8), ED(50) 28.1 mg kg(-1) po in rats, TD(50) > 500 mg kg(-1), PI > 17.8. Sodium channel binding studies, as well as evaluations against pentylenetetrazol, bicuculline, and picrotoxin on isoxazole 10 were all negative, leading to an unknown mechanism of action. X-ray diffraction patterns of a representative of the 3-amino series (isoxazoles 6-11) unequivocally display the existence of intramolecular hydrogen bonding of the nitrogen to the vinylic proton in the cyclohexene ring, providing a pseudo three ring structure which was also shown previously with the vinylic benzamides. Physicochemical-permeability across the BBB suggested an efflux mechanism for the previously synthesised aniline enaminones, but not with isoxazole 10. (C) 2002 Published by Editions scientifiques et medicales Elsevier SAS. C1 Howard Univ, Sch Pharm, Dept Pharmaceut Sci, Washington, DC 20059 USA. Univ Maryland, Sch Pharm, Dept Pharmaceut Sci, Baltimore, MD 21201 USA. DuPont Pharmaceut, Drug Metab & Pharmacokinet Sect, Newark, DE 19713 USA. 3M Co, Corp Res, 3M Ctr, Sci Res Lab, St Paul, MN 55144 USA. Howard Univ, Grad Sch, Dept Chem, Washington, DC 20059 USA. Kuwait Univ, Fac Pharm, Dept Pharmaceut Chem, Jabriya, Kuwait. NINDS, Epilepsy Branch, Div Convuls Dev & Neuromuscular Disorders, Bethesda, MD 20892 USA. RP Scott, KR (reprint author), Howard Univ, Sch Pharm, Dept Pharmaceut Sci, Washington, DC 20059 USA. EM kscott@fac.howard.edu NR 37 TC 84 Z9 85 U1 2 U2 10 PU ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS PA 23 RUE LINOIS, 75724 PARIS, FRANCE SN 0223-5234 J9 EUR J MED CHEM JI Eur. J. Med. Chem. PD AUG PY 2002 VL 37 IS 8 BP 635 EP 648 AR PII S0223-5234(02)01377-6 DI 10.1016/S0223-5234(02)01377-6 PG 14 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 592LP UT WOS:000177939400002 PM 12161061 ER PT J AU Mead, AN Rocha, BA Donovan, DM Katz, JL AF Mead, AN Rocha, BA Donovan, DM Katz, JL TI Intravenous cocaine induced-activity and behavioural sensitization in norepinephrine-, but not dopamine-transporter knockout mice SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE DAT; KO mice; NET; nucleus accumbens ID VENTRAL TEGMENTAL AREA; FREELY MOVING RATS; EXTRACELLULAR DOPAMINE; NUCLEUS-ACCUMBENS; INTRACEREBRAL DIALYSIS; AMPHETAMINE; LOCOMOTOR; LACKING; PSYCHOSTIMULANTS; SCH-23390 AB Previously, it was reported that both norepinephrine transporter (NET) and dopamine transporter (DAT) knockout (KO) mice were sensitive to the reinforcing effects of cocaine. However, assessing the locomotor-stimulant effects of cocaine in these subjects has proven difficult due to significant differences in their baseline activity compared to wild-type controls. The present studies were designed to clarify the role of NET and DAT in the stimulant effects of acute and repeated cocaine utilizing these knockout mice, and thereby assess the role of these substrates in the locomotor stimulant effects of cocaine. Mice were habituated to the test environment for sufficient time to ensure equal baselines at the time of cocaine administration. Mice then received cocaine (3-25 mg/kg) intravenously according to a within-session cumulative dose-response design. Cocaine dosing was repeated at 48-h intervals for four sessions to assess behavioural sensitization. NET-KO mice exhibited a reduced response to acute cocaine administration compared to wild-type (WT) controls. However, comparable sensitization developed in NET-KO and WT mice. The DAT-KO and DAT-heterozygote (HT) mice displayed no locomotor activation following either acute or repeated cocaine administration. These data suggest a role for the NET in the acute response to cocaine, but no involvement in sensitization to cocaine. In contrast, DAT appears to be necessary for both the acute locomotor response to cocaine and the subsequent development of sensitization. In addition to existing data concerning the reinforcing effects of cocaine in DAT-KO mice, these data suggest a dissociation between the reinforcing and locomotor stimulant effects of cocaine. C1 NIDA, Behav Neurosci Branch, Intramural Res Program, Baltimore, MD 21224 USA. NIDA, Med Discovery Res Branch, Intramural Res Program, Baltimore, MD 21224 USA. NIA, Gerontol Res Ctr, IRP, NIH, Baltimore, MD 21224 USA. Univ Maryland, Maryland Psychiat Res Ctr, Dept Psychiat, Catonsville, MD 21228 USA. RP Mead, AN (reprint author), Univ Sussex, Sch Biol Sci, Expt Psychol Lab, Brighton BN1 9QG, E Sussex, England. OI Katz, Jonathan/0000-0002-1068-1159 FU NIDA NIH HHS [R29 DA 12579] NR 27 TC 33 Z9 33 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD AUG PY 2002 VL 16 IS 3 BP 514 EP 520 DI 10.1046/j.1460-9568.2002.02104.x PG 7 WC Neurosciences SC Neurosciences & Neurology GA 586QU UT WOS:000177596500022 PM 12193195 ER PT J AU Sridhar, SK Pandeya, SN Stables, JP Ramesh, A AF Sridhar, SK Pandeya, SN Stables, JP Ramesh, A TI Anticonvulsant activity of hydrazones, Schiff and Mannich bases of isatin derivatives SO EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES LA English DT Article DE isatin; hydrazones; Schiff base; Mannich base; anticonvulsant ID ANTIEPILEPTIC DRUG DEVELOPMENT; SEMICARBAZONES AB In the present study, anticonvulsant activity of hydrazones, Schiff and Mannich bases of isatin were evaluated by maximal electroshock method (MES) and metrazol-induced convulsions (MET) at 30, 100 and 300 mg/kg dose levels. Neurotoxicity of the compounds was also assessed at the same dose levels. Eight compounds of the series exhibited significant anticonvulsant activity at 30 mg/kg dose level. 3-(4-chloro-phenylimino)-5-methyl-1,3-dihydro-indol-2-one (compound 10) was found to be the most potent compound of the series with 87% protection at 100 mg/kg and an ED50 of 53.61 mg/kg (MET). All the compounds exhibited lesser neurotoxicity compared to phenytoin. All the active compounds showed greater protection than sodium valproate. The essential structural features responsible for interaction with receptor site are established within a suggested pharmacophore. (C) 2002 Elsevier Science B.V. All rights reserved. C1 Fredrick Inst Plant Protect & Toxicol, Dept Pesticide Chem, Padappai 601301, India. Banaras Hindu Univ, Inst Technol, Dept Pharmaceut, Varanasi 221005, Uttar Pradesh, India. NIH, Preclin Pharmacol Sect, Epilepsy Branch, Bethesda, MD 20892 USA. RP Ramesh, A (reprint author), Fredrick Inst Plant Protect & Toxicol, Dept Pesticide Chem, Padappai 601301, India. NR 27 TC 151 Z9 157 U1 1 U2 12 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0928-0987 J9 EUR J PHARM SCI JI Eur. J. Pharm. Sci. PD AUG PY 2002 VL 16 IS 3 BP 129 EP 132 AR PII S0928-0987(02)00077-5 DI 10.1016/S0928-0987(02)00077-5 PG 4 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 582EY UT WOS:000177337800005 PM 12128166 ER PT J AU Weinberger, DR AF Weinberger, DR TI Schizophrenia, the prefrontal cortex, and a mechanism of genetic susceptibility SO EUROPEAN PSYCHIATRY LA English DT Article; Proceedings Paper CT Mind Matters International Schizophrenia Fourm CY JUN, 2001 CL BARCELONA, SPAIN DE schizophrenia; executive cognitive function; dopamine; COMT; genetic risk ID D1 RECEPTORS; RAT AB Information-processing in the prefrontal cortex is abnormal inpatients with schizophrenia. From functional neuroimaging and other studies, it appears that neurons of the dorsolateral prefrontal cortex are effectors of core clinical and biological phenomena associated with the illness. Cognitive deficits qualitatively similar to those in patients with schizophrenia are also found in their healthy siblings and other relatives. The evidence for abnormal prefrontal function in healthy siblings suggests that the deficit in informal ion-processing is part of the biology of genetic susceptibility. Therefore, genetic variations in human DNA that affect this kind of information-processing may contribute part of the genetic risk for the illness. COMT is an enzyme that is distributed widely throughout the brain, but seems to be uniquely relevant to how dopamine affects information-processing in the prefrontal cortex. There is a common variation in the genetic sequence of the COMT gene, which causes a dramatic change in its enzyme activity. In people with schizophrenia, in their healthy siblings, and also in normal controls, the COMT genotype predicts 4% of the variation in human executive cognition and working memory. We cave thus identified a genetic mechanism in the human species that affects the efficiency and efficacy of information-processing in the prefrontal cortex. It is, also, a weak genetic risk factor for schizophrenia: in family studies, it increases the risk of schizophrenia by 50-80%. Prefrontal neuronal tuning may be a target for new drug development in the future. (C) 2002 Editions scientifiques et medicales Elsevier SAS. C1 NIMH, Clin Brain Disorders Branch, Intramural Res Program, NIH, Bethesda, MD 20892 USA. RP Weinberger, DR (reprint author), NIMH, Clin Brain Disorders Branch, Intramural Res Program, NIH, 10 Ctr Dr, Bethesda, MD 20892 USA. NR 16 TC 7 Z9 7 U1 0 U2 4 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 0924-9338 J9 EUR PSYCHIAT JI Eur. Psychiat. PD AUG PY 2002 VL 17 SU 4 BP 355 EP 362 PG 8 WC Psychiatry SC Psychiatry GA 593MV UT WOS:000177996000003 ER PT J AU Leshner, AI AF Leshner, AI TI Experimental and clinical psychopharmacology: National Institute on Drug Abuse's clinical research agenda SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material AB Studies of drugs and behavior are a core component of virtually every portfolio within the broad purview of the National Institute on Drug Abuse (NIDA). Moreover, psychopharmacological research is an important vehicle for advancing understanding of how drugs of abuse produce their effects, particularly including addiction. However, as with all major public health issues, simply understanding the issue is not enough. NIDA's psychopharmacology projects, therefore, span basic, clinical, and applied (e.g., medication development) research activities. These include the establishment of a nationwide clinical trials network designed to provide an infrastructure to test both behavioral and psychopharmacological treatments in a real-life practice setting with diverse patients. C1 NIDA, Bethesda, MD 20892 USA. RP Leshner, AI (reprint author), Amer Assoc Advancement Sci, 1200 New York Ave NW, Washington, DC 20005 USA. NR 0 TC 8 Z9 8 U1 0 U2 1 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD AUG PY 2002 VL 10 IS 3 BP 159 EP 161 DI 10.1037//1064-1297.10.3.159 PG 3 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 588QJ UT WOS:000177711900002 PM 12233978 ER PT J AU Cristillo, AD Xiao, XD Campbell-Malone, R Dimitrov, DS Bierer, BE AF Cristillo, AD Xiao, XD Campbell-Malone, R Dimitrov, DS Bierer, BE TI Differences between CEM and human peripheral blood T lymphocytes in cAMP-dependent HIV viral fusion and CXCR4 expression SO EXPERIMENTAL AND MOLECULAR PATHOLOGY LA English DT Article DE CXCR4; HIV; cAMP; CREB; chemokine; chemokine receptors; AIDS ID IMMUNODEFICIENCY-VIRUS TYPE-1; CHEMOKINE RECEPTOR CXCR4; HEMATOPOIETIC PROGENITOR CELLS; SURFACE EXPRESSION; ENTRY; INFECTION; SDF-1; CORECEPTOR; CHEMOATTRACTANT; CLONING AB CXCR4, a G-protein-coupled chemokine receptor and HIV coreceptor, has been shown to play a central role in both chemotaxis and HIV-1-entry into T lymphocytes. Recent efforts have focused on identifying the signaling pathways that modulate CXCR4 expression in order to modulate HIV infectivity. Toward this effort, we previously demonstrated cAMP-dependent up-regulation of CXCR4 mRNA and protein in human peripheral blood T cells (PBL), resulting in increased HIV infectivity. Regulation of CXCR4 mRNA was mediated, in part, by a CRE element within the CXCR4 promoter. In order to develop a model system to examine cAMP regulation, the responses of the T lymphoblastoid cell line CEM were compared to those of human PBL. In sharp contrast to that of human PBL, HIV-1 entry into CEM cells was dramatically reduced in response to dibutyryl cAMP (DcAMP). Furthermore, while total cellular and cell surface CXCR4 protein levels were up-regulated in human PBL and in Jurkat T cells in response to DcAMP or forskolin stimulation, CXCR4 levels were unchanged by stimulation in CEM cells. Surprisingly, the CXCR4 promoter (nucleotides - 1098 to +59) fused to luciferase was found to be activated similarly in CEM and Jurkat cells in response to DcAMP in a concentration-dependent manner. RT-PCR analyses confirmed that CXCR4 mRNA levels were increased by cAMP agonists. Taken together, our findings suggest that total and cell surface CXCR4 protein expression is regulated differently in human PBL than in CEM cells, a finding that correlates with the differential HIV-1 fusion in response to cAMP signaling. Moreover, our results suggest that, for CXCR4 expression and HIV viral infectivity, CEM cells may not be a faithful model of primary human lymphocytes. (C) 2002 Elsevier Science (USA). C1 NHLBI, Lab Lymphocyte Biol, NIH, Bethesda, MD 20892 USA. NCI, FCRDC, NIH, Bethesda, MD 20892 USA. RP Bierer, BE (reprint author), NHLBI, Lab Lymphocyte Biol, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 39 TC 9 Z9 9 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4800 J9 EXP MOL PATHOL JI Exp. Mol. Pathol. PD AUG PY 2002 VL 73 IS 1 BP 9 EP 18 DI 10.1006/exmp.2002.2443 PG 10 WC Pathology SC Pathology GA 572TF UT WOS:000176790500002 PM 12127049 ER PT J AU Hematti, P Sloand, EM Carvallo, CA Albert, MR Yee, CL Fuehrer, MM Blancato, JK Kearns, WG Barrett, JA Childs, RW Vogel, JC Dunbar, CE AF Hematti, P Sloand, EM Carvallo, CA Albert, MR Yee, CL Fuehrer, MM Blancato, JK Kearns, WG Barrett, JA Childs, RW Vogel, JC Dunbar, CE TI Absence of donor-derived keratinocyte stem cells in skin tissues cultured from patients after mobilized peripheral blood hematopoietic stem cell transplantation SO EXPERIMENTAL HEMATOLOGY LA English DT Article ID BONE-MARROW; IN-VIVO; ADULT MICE; EXPRESSION; BRAIN; DIFFERENTIATE; HEPATOCYTES; ENGRAFTMENT; PROGENITORS; SURVIVAL AB Objective. Recent studies suggest that primitive bone marrow-derived cells contribute to regeneration of many tissues, including muscle, endothelium, myocardium, neural tissues, liver, and skin. Conversely, primitive cells. resident in muscle and other tissues have been reported to reconstitute hematopoiesis. We investigated the contribution of cells with a primitive hematopoietic phenotype to human epidermal skin formation in recipients of allogeneic mobilized peripheral blood hematopoietic stem cell (HSC) transplantation. Patients and Methods. Our study population included female patients who had received granulocyte colony-stimulating factor mobilized peripheral blood HSC transplants from male donors for a variety of benign and malignant hematologic disorders at least 6 months before study entry, with a history of skin graft-vs-host disease. Epidermal skin cells (keratinocytes) obtained from punch biopsies of the skin were cultured under conditions specific for growth and expansion of homogenous populations of keratinocytes from keratinocyte stem cells. After multiple passages, DNA was extracted from cultured cells and evaluated by two different polymerase chain reaction (PCR) method for detection of Y chromosome specific sequences. Results. Neither sensitive PCR-based technique revealed the presence of male donor-derived keratinocyte stem cells in keratinocytes cultured from skin biopsies of female allogeneic transplantation recipients. Conclusions. We could not confirm the contribution of donor mobilized peripheral blood hematopoietic stem cells to keratinocyte stem cell populations after HSC transplantation. These results cannot explain the presence of donor-derived cells with keratinocyte phenotypic markers in tissue sections of HSC transplant recipients. (C) 2002 International Society for Experimental Hematology. Published by Elsevier Science Inc. C1 NIH, NHLBI, Hematol Branch, Bethesda, MD 20892 USA. NIH, Urol Oncol Branch, NCI, Bethesda, MD USA. NIH, Dermatol Branch, NCI, Bethesda, MD USA. Georgetown Univ, Ctr Med, Inst Mol & Human Genet, Washington, DC USA. RP Dunbar, CE (reprint author), NIH, NHLBI, Hematol Branch, Bldg 10,Rm 7C103,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 32 TC 34 Z9 36 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0301-472X J9 EXP HEMATOL JI Exp. Hematol. PD AUG PY 2002 VL 30 IS 8 BP 943 EP 949 AR PII S0301-472X(02)00873-1 DI 10.1016/S0301-472X(02)00873-1 PG 7 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 586WZ UT WOS:000177610300013 PM 12160846 ER PT J AU Wu, CG Forgues, M Siddique, S Farnsworth, J Velerie, K Wang, XW AF Wu, CG Forgues, M Siddique, S Farnsworth, J Velerie, K Wang, XW TI SAGE transcript profiles of normal primary human hepatocytes expressing oncogenic hepatitis B virus X protein SO FASEB JOURNAL LA English DT Article DE liver cancer; Megarray; nuclear export signal ID GENE-EXPRESSION; HEPATOCELLULAR-CARCINOMA; TRANSGENIC MICE; SERIAL ANALYSIS; LIVER-CANCER; HEPATOCARCINOGENESIS; INFECTION; LOCALIZATION; ETIOLOGY; COMPLEX AB Hepatitis B virus (HBV) is a major risk factor for hepatocellular carcinoma (HCC). HBV encodes an oncogenic HBx gene that functions as a transcriptional coactivator of multiple cellular genes. To understand the role(s) of HBx in the early genesis of HCC, we systematically analyzed gene expression profiles by serial analysis of gene expression (SAGE) in freshly isolated human primary hepatocytes infected with a replication-defective adenovirus containing HBx. A total of 19,501 sequence tags (representing 1443 unique transcripts) were analyzed, which provide a distribution of a transcriptome characteristic of normal hepatocytes and a profile associated with HBx expression. Examples of the targeted genes were confirmed by the Megarray analysis with a significant correlation between quantitative SAGE and Megarray (r = 0.8, P < 0.005). In HBx-expressing hepatocytes, a total of 57 transcripts (3.9%) were induced, and 46 transcripts (3.3%) were repressed by more than fivefold. Interestingly, most of the HBx-up-regulated transcripts can be clustered into three major classes, including genes that encode ribosomal proteins, transcription factors with zinc-finger motifs, and proteins associated with protein degradation pathways. These results suggest that HBx may function as a major regulator in common cellular pathways that, in turn, regulate protein synthesis, gene transcription, and protein degradation. C1 NCI, Human Carcinogenesis Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA. US FDA, Div Hematol, Ctr Biol Evaluat & Res, Bethesda, MD 20014 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Radiat Oncol, Richmond, VA 23298 USA. RP Wang, XW (reprint author), NCI, Human Carcinogenesis Lab, Ctr Canc Res, NIH, 37 Convent Dr,MSC 4255,Bldg 37,Room 2C25, Bethesda, MD 20892 USA. EM xin_wei_wang@nih.gov RI Wang, Xin/B-6162-2009 NR 39 TC 25 Z9 28 U1 0 U2 0 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD AUG PY 2002 VL 16 IS 10 BP 1665 EP + DI 10.1096/fj.02-0074fje PG 24 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 590HQ UT WOS:000177814000019 PM 12207007 ER PT J AU Scott, RT Levi, A AF Scott, RT Levi, A TI An experimental bagatelle or a wrong mistake? Reply SO FERTILITY AND STERILITY LA English DT Letter C1 Reprod Med Associates New Jersey, Morristown, NJ USA. NICHHD, Pediat & Reprod Endocrine Branch, Bethesda, MD 20892 USA. RP Scott, RT (reprint author), Reprod Med Associates New Jersey, Morristown, NJ USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0015-0282 J9 FERTIL STERIL JI Fertil. Steril. PD AUG PY 2002 VL 78 IS 2 BP 435 EP 436 AR PII S0015-0282(02)03256-9 DI 10.1016/S0015-0282(02)03256-9 PG 2 WC Obstetrics & Gynecology; Reproductive Biology SC Obstetrics & Gynecology; Reproductive Biology GA 581JF UT WOS:000177287900039 ER PT J AU Detweiler, CD Deterding, LJ Tomer, KB Chignell, CF Germolec, D Mason, RP AF Detweiler, CD Deterding, LJ Tomer, KB Chignell, CF Germolec, D Mason, RP TI Immunological identification of the heart myoglobin radical formed by hydrogen peroxide SO FREE RADICAL BIOLOGY AND MEDICINE LA English DT Article DE myoglobin; 5,5-dimethyl-1-pyrroline N-oxide (DMPO); antibodies; protein radical; western blot; free radicals ID MASS-SPECTROMETRY; CYTOCHROME-C; METMYOGLOBIN; REPERFUSION; ADDUCT; ISCHEMIA AB This study reports the detection of protein free radicals using the specific free radical reactivity of nitrone spin traps in conjunction with nitrone-antibody specificity. Polyclonal antibodies were developed that bind to protein adducts of the nitrone spin-trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO). The antibodies were used to detect DMPO protein adducts produced on horse myoglobin resulting from self-peroxidation. Western blot analysis demonstrates that myoglobin forms the predominant radical-derived nitrone adduct in rat heart Supernatant. Published by Elsevier Science Inc. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Mason, RP (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233,MD F001, Res Triangle Pk, NC 27709 USA. RI Tomer, Kenneth/E-8018-2013 NR 21 TC 96 Z9 97 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0891-5849 J9 FREE RADICAL BIO MED JI Free Radic. Biol. Med. PD AUG 1 PY 2002 VL 33 IS 3 BP 364 EP 369 AR PII S0891-5849(02)00895-X DI 10.1016/S0891-5849(02)00895-X PG 6 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 577XU UT WOS:000177089000008 PM 12126758 ER PT J AU Cobb, S Wood, T Tessarollo, L Velasco, M Given, R Varro, A Tarasova, N Singh, P AF Cobb, S Wood, T Tessarollo, L Velasco, M Given, R Varro, A Tarasova, N Singh, P TI Deletion of functional gastrin gene markedly increases colon carcinogenesis in response to azoxymethane in mice SO GASTROENTEROLOGY LA English DT Article ID GLYCINE-EXTENDED GASTRIN; HELICOBACTER-PYLORI INFECTION; HORMONE REPLACEMENT THERAPY; ESTROGEN-RECEPTOR-BETA; MALIGNANT HUMAN-COLON; CANCER-CELLS INVITRO; COLORECTAL-CANCER; TRANSGENIC MICE; MESSENGER-RNA; GROWTH-FACTOR AB Background & Aims: We recently reported that transgenic mice overexpressing progastrin were at a higher risk for developing colon cancers in response to azoxymethane (AOM), whereas mice overexpressing gastrin-17 were at a reduced risk. To examine further the role of gastrins in colon carcinogenesis, we generated gastrin gene knockout mice (GAS-KO). Methods: The height and proliferative index (PI) of colonic crypts were similar in GAS-KO and wild-type (WT) mice, suggesting that the absence of gastrins in GAS-KO mice did not significantly affect the growth of colonic mucosa. GAS-KO and WT mice were treated with AOM for 3-4 weeks; control mice received saline. Results: Colonic proliferation in response to AOM was significantly increased in GAS-KO vs. WT mice. Aberrant crypt foci (ACFs) were similarly increased significantly by similar to2-5-fold in GAS-KO vs. WT mice after 2 weeks of AOM treatment. Female GAS-KO mice developed adenomas (Ads) and adenocarcinomas (AdCAs) at earlier times (similar to10 months) than the male GAS-KO mice and the male and female WT mice (similar to12 months). The total numbers of Ads and AdCAs were significantly higher in GAS-KO than in WT mice. Conclusions: These results suggest the novel possibility that loss of gastrin expression (and hence amidated gastrins) significantly increases susceptibility to colon carcinogenesis in response to AM Previous studies with FVB/N transgenic mice similarly suggested a protective role of amidated gastrins against colon carcinogenesis, which supports the present findings of an increase in colon carcinogenesis in GAS-KO mice lacking normal physiological levels of amidated gastrins. C1 Univ Texas, Med Branch, Dept Anat & Neurosci, Galveston, TX 77555 USA. Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77550 USA. NCI, Fredericksburg, MA USA. Vel Lab Res, Houston, TX USA. Univ Liverpool, Dept Physiol, Liverpool L69 3BX, Merseyside, England. RP Singh, P (reprint author), Univ Texas, Med Branch, Dept Anat & Neurosci, Galveston, TX 77555 USA. RI Wood, Thomas/B-6172-2012; Varro, Andras/M-2647-2016 OI Varro, Andras/0000-0003-0745-3603 FU NCI NIH HHS [CA60087, CA72992] NR 69 TC 26 Z9 26 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD AUG PY 2002 VL 123 IS 2 BP 516 EP 530 DI 10.1053/gast.2002.34754 PG 15 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 577WK UT WOS:000177085400016 PM 12145805 ER PT J AU Bouma, G Kaushiva, A Strober, W AF Bouma, G Kaushiva, A Strober, W TI Experimental murine colitis is regulated by two genetic loci, including one on chromosome 11 that regulates IL-12 responses SO GASTROENTEROLOGY LA English DT Article ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; CROHNS-DISEASE; SUSCEPTIBILITY LOCUS; INTERLEUKIN-12; POLYMORPHISMS; AUTOIMMUNITY; COMPLEX; CELLS AB Background & Aims: Immunogenetic analysis of experimental colitis may contribute to the further unraveling of the complex genetic basis of the inflammatory bowel diseases, Crohn's disease and ulcerative colitis. Methods: Genetic regions associated with susceptibility to trinitrobenzene sulfonic acid (TNBS)-induced colitis were identified in a genome-wide linkage analysis in F2 progeny of colitis-susceptible SJL/J and -resistant C57BL/6 mice. An immunogenetic approach was then used to further study the pathophysiologic role of one of the identified loci. Results: We identified susceptibility loci on chromosomes 9 (Tnbs1) and 11 (Tnbs2). Tnbs2 harbors the interleukin (IL)-12 p40 gene, a likely candidate gene because IL-12 is a known central mediator for both experimental colitis and human Crohn's disease. We therefore tested the ability of colitis-susceptible and -resistant strains to mount IL-12 responses to lipopolysaccharide (LPS), a strong inducer of IL-12 that is abundantly present in the intestine. We observed a remarkably higher serum IL-12 response to LPS in susceptible SJL/J mice. Subsequently, we showed that the genetic region regulating the IL-12 response to LPS colocalizes with Tnbs2. Conclusions: These data strongly suggest that the tendency to mount a high LPS-induced IL-12 response and susceptibility to TNBS-induced colitis are related and that in fact a genetically determined high IL-12 response is involved in the immunologic basis of susceptibility to colitis. C1 NIAID, Mucosal Immun Sect, NIH, Bethesda, MD 20892 USA. Vrije Univ Amsterdam, Ctr Med, Lab Gastrointestinal Immunogenet, Amsterdam, Netherlands. RP Strober, W (reprint author), NIAID, Mucosal Immun Sect, NIH, Bldg 10,11N238, Bethesda, MD 20892 USA. RI bouma, gerd/E-2520-2013 NR 25 TC 54 Z9 55 U1 0 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD AUG PY 2002 VL 123 IS 2 BP 554 EP 565 DI 10.1053/gast.2002.34752 PG 12 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 577WK UT WOS:000177085400019 PM 12145808 ER PT J AU Hanazono, Y Nagashima, T Takatoku, M Shibata, H Ageyama, N Asano, T Ueda, Y Dunbar, CE Kume, A Terao, K Hasegawa, M Ozawa, K AF Hanazono, Y Nagashima, T Takatoku, M Shibata, H Ageyama, N Asano, T Ueda, Y Dunbar, CE Kume, A Terao, K Hasegawa, M Ozawa, K TI In vivo selective expansion of gene-modified hematopoietic cells in a nonhuman primate model SO GENE THERAPY LA English DT Article DE hematopoietic stem cell; gene therapy; retroviral vector; selective amplifier gene; in vivo expansion; nonhuman primate; transplantation ID BONE-MARROW CELLS; COLONY-STIMULATING FACTOR; RHESUS PERIPHERAL-BLOOD; STEM-CELLS; IN-VIVO; RETROVIRAL VECTORS; AMPLIFIER GENE; TRANSPLANTED MICE; MEDIATED TRANSFER; BREAST-CANCER AB A major problem limiting hematopoietic stem cell (HSC) gene therapy is the low efficiency of gene transfer into human HSCs using retroviral vectors. Strategies, which would allow in vivo expansion of gene-modified hematopoietic cells, could circumvent the problem. To this end, we developed a selective amplifier gene (SAG) consisting of a chimeric gene composed of the granulocyte colony-stimulating factor (G-CSF) receptor gene and the estrogen receptor gene hormone-binding domain. We have previously demonstrated that primary bone marrow progenitor cells transduced with the SAG could be expanded in response to estrogen in vitro. In the present study, we evaluated the efficacy of the SAG in the setting of a clinically applicable cynomolgus monkey transplantation protocol. Cynomolgus bone marrow CD34(+) cells were transduced with retroviral vectors encoding the SAG and reinfused into each myeloablated monkey. Three of the six monkeys that received SAG transduced HSCs showed an increase in the levels of circulating progeny containing the provirus in vivo following administration of estrogen or tamoxifen without any serious adverse effects. In one monkey examined in detail, transduced hematopoietic progenitor cells were increased by several-fold (from 5% to 30%). Retroviral integration site analysis revealed that this observed increase was polyclonal and no outgrowth of a dominant single clonal population was observed. These results demonstrate that the inclusion of our SAG in the retroviral construct allows selective in vivo expansion of genetically modified cells by a non-toxic hormone treatment. C1 Jichi Med Sch, Div Genet Therapeut, Ctr Mol Med, Kawachi, Tochigi 3290498, Japan. Jichi Med Sch, Div Hematol, Dept Med, Kawachi, Tochigi 3290498, Japan. DNAVEC Res Inc, Ibaraki, Japan. Natl Inst Infect Dis, Tsukuba Primate Ctr, Ibaraki, Japan. NHLBI, Hematol Branch, Bethesda, MD USA. RP Ozawa, K (reprint author), Jichi Med Sch, Div Genet Therapeut, Ctr Mol Med, 3311-1 Yakushiji, Kawachi, Tochigi 3290498, Japan. NR 48 TC 31 Z9 32 U1 0 U2 5 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0969-7128 J9 GENE THER JI Gene Ther. PD AUG PY 2002 VL 9 IS 16 BP 1055 EP 1064 DI 10.1038/sj.gt.3301781 PG 10 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 577WM UT WOS:000177085600001 PM 12140733 ER PT J AU Royal, RE Kershaw, MH Reeves, ME Wang, G Daly, T Treisman, J Lam, J Hwu, P AF Royal, RE Kershaw, MH Reeves, ME Wang, G Daly, T Treisman, J Lam, J Hwu, P TI Increased functional expression of transgene in primary human lymphocytes using retroviral vectors modified with IRES and splicing motifs SO GENE THERAPY LA English DT Article DE immunotherapy; colon cancer; lymphocyte; retrovirus; vectors ID TUMOR-INFILTRATING LYMPHOCYTES; RIBOSOMAL ENTRY SITE; CYTOTOXIC T-LYMPHOCYTES; MEDIATED GENE-TRANSFER; APE LEUKEMIA-VIRUS; ENCEPHALOMYOCARDITIS VIRUS; IN-VIVO; CELLS; THERAPY; EFFICIENCY AB Genetic modification of human lymphocytes is being employed in strategies to correct enzyme deficiencies, encode cytokines and to redirect lymphocytes to antigenic targets other than those encoded by their endogenous T cell receptor. However, expression of transgenes in primary lymphocytes is generally low, Reasoning that vector modification may lead to increased transgene expression and subsequent increases in function, we have performed two retroviral vector modifications and report their effect on the functional expression in primary lymphocytes. A chimeric receptor specific for the colon carcinoma-associated antigen, EGP40, was initially incorporated into the retroviral vector LXSN. In this vector, receptor expression is driven by the Moloney murine leukemia virus LTR, and neomycin phosphotransferase expression driven by the SV40 promoter. Replacement of SV40 with an internal ribosomal entry site (IRES) increased the transgene activity of a mouse T cell line and human PBL as judged by increased cytokine release in response to antigen positive target cells. A further increase in transgene function was generated by the additional incorporation of a splice acceptor motif into the construct. Human PBL transduced with vector incorporating both IRES and intron were consistently more effective at lysing antigen positive colorectal carcinoma cells. C1 NCI, Tumor Immunol Sect, Canc Res Ctr, NIH, Bethesda, MD 20892 USA. RP Hwu, P (reprint author), NCI, Tumor Immunol Sect, Canc Res Ctr, NIH, Bldg 10,Room 2B42,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Kershaw, Michael/0000-0002-2697-487X NR 39 TC 12 Z9 13 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0969-7128 J9 GENE THER JI Gene Ther. PD AUG PY 2002 VL 9 IS 16 BP 1085 EP 1092 DI 10.1038/sj.gt.3301734 PG 8 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 577WM UT WOS:000177085600004 PM 12140736 ER PT J AU Klein, AP Beaty, TH Bailey-Wilson, JE Brune, KA Hruban, RH Petersen, GM AF Klein, AP Beaty, TH Bailey-Wilson, JE Brune, KA Hruban, RH Petersen, GM TI Evidence for a major gene influencing risk of pancreatic cancer SO GENETIC EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT 10th Annual International-Genetic-Epidemiology-Society Meeting CY SEP, 2001 CL GARMISCH PARTENKI, GERMANY SP Int Genet Epidemiol Soc DE segregation analysis; pancreatic cancer ID SEGREGATION ANALYSIS; FAMILY HISTORY; CARCINOMA; ADENOCARCINOMA; MANAGEMENT; LOUISIANA; BROTHERS AB Family history of pancreatic cancer, the fifth leading cause of cancer death in the United States, confers a 1.5-13-fold higher risk of developing pancreatic cancer. Pancreatic cancer is associated with several genetic syndromes, including hereditary breast cancer (BRCA2), familial atypical multiple mole melanoma (FAMMM) syndrome, Peutz-Jeghers syndrome, hereditary pancreatitis, and hereditary nonpolyposis colorectal cancer (HNPCC. However, these syndromes explain little of the observed familial aggregation of pancreatic cancer. We performed complex segregation analysis on 287 families ascertained through an index case diagnosed with pancreatic cancer at the Johns Hopkins Medical Institutions between January 1, 1994 and December 31, 1999. We tested for the presence of a major gene controlling either the "age-at-onset of pancreatic cancer" of "susceptibility to pancreatic cancer," and incorporated smoking data on kindred members as a covariate. We found evidence for involvement of a major gene in the etiology of pancreatic cancer. Whether inheritance was modeled as "age-at-onset" or "susceptibility," nongenetic transmission models were strongly rejected. However, modeling "age-at-onset" provided a better fit to the observed data than did modeling "susceptibility." The most parsimonious models included autosomal-dominant inheritance of a rare allele. Under the age-at-onset model, approximately 0.7% of the population appears to be at high risk of developing pancreatic cancer due to this putative gene, whereas 0.4% of the population is at high risk under the susceptibility model. Inclusion of smoking as a covariate did not significantly improve the fit of these models. This hospital-based segregation analysis of pancreatic cancer found evidence supporting the role of a rare major gene influencing risk of pancreatic cancer. Genet. Epidemiol. 23:133-149, 2002. (C) 2002 Wiley-Liss, Inc. C1 Mayo Clin, Dept Hlth Sci Res, Rochester, MN 55905 USA. Johns Hopkins Univ, Dept Epidemiol, Bloomberg Sch Publ Hlth, Baltimore, MD USA. Natl Human Genome Res Inst, NIH, Sect Stat Genet, Inherited Dis Res Branch, Baltimore, MD USA. Johns Hopkins Univ, Dept Pathol, Sch Med, Baltimore, MD USA. Johns Hopkins Univ, Sch Med, Dept Oncol, Baltimore, MD USA. RP Petersen, GM (reprint author), Mayo Clin, Dept Hlth Sci Res, 200 1st St,SW, Rochester, MN 55905 USA. OI Bailey-Wilson, Joan/0000-0002-9153-2920 FU NCI NIH HHS [P50-CA62924, R03 CA88273]; NCRR NIH HHS [1 P41 RR03655] NR 38 TC 81 Z9 83 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0741-0395 J9 GENET EPIDEMIOL JI Genet. Epidemiol. PD AUG PY 2002 VL 23 IS 2 BP 133 EP 149 DI 10.1002/gepi.01102 PG 17 WC Genetics & Heredity; Mathematical & Computational Biology SC Genetics & Heredity; Mathematical & Computational Biology GA 588NP UT WOS:000177707800004 PM 12214307 ER PT J AU Aravind, L Iyer, LM AF Aravind, L Iyer, LM TI Intraproteomic networks: New forays into predicting interaction partners SO GENOME RESEARCH LA English DT Editorial Material ID DNA-BINDING; LEUCINE ZIPPER; COILED COILS; SANT DOMAIN; PROTEIN; COMPLEX; IDENTIFICATION; EVOLUTION; MOTIF C1 Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RP Aravind, L (reprint author), Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. EM aravind@ncbl.nlm.nih.gov NR 18 TC 0 Z9 0 U1 0 U2 0 PU COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT PI COLD SPRING HARBOR PA 1 BUNGTOWN RD, COLD SPRING HARBOR, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD AUG PY 2002 VL 12 IS 8 BP 1156 EP 1158 DI 10.1101/gr.353302 PG 3 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 583WQ UT WOS:000177434300002 PM 12176922 ER PT J AU Fassler, J Landsman, D Acharya, A Moll, JR Bonovich, M Vinson, C AF Fassler, J Landsman, D Acharya, A Moll, JR Bonovich, M Vinson, C TI B-ZIP proteins encoded by the Drosophila genome: Evaluation of potential dimerization partners SO GENOME RESEARCH LA English DT Article ID HELICAL COILED COILS; DNA-BINDING PROTEIN; GCN4 LEUCINE-ZIPPER; TRANSCRIPTION FACTORS; CRYSTAL-STRUCTURE; BASIC REGION; EMBRYONIC-DEVELOPMENT; NUCLEAR-PROTEIN; BZIP PROTEIN; AMINO-ACID AB The basic region-leucine zipper (bZIP) (bZIP) protein motif dimerizes to bind specific DNA Sequences. We have identified 27 B-ZIP proteins in the recently sequenced Drosophila melanogaster genome. The dimerization specificity of these 27 B-ZIP proteins was evaluated using two structural criteria: (1) the presence of attractive or repulsive interhelical g<---->e' electrostatic interactions and (2) the presence of polar or charged amino acids in the 'a' and V positions of the hydrophobic interface. None of the B-ZIP proteins contain only aliphatic amino acids in the 'a' and 'd' position. Only six of the Drosophila B-ZIP proteins contain a "canonical" hydrophobic interface like the yeast GCN4, and the mammalian JUN, ATF2, CREB, C/EBP, and PAR leucine zippers, characterized by asparagine in the second 'a' position. Twelve leucine zippers contain polar amino acids in the first, third, and fourth 'a' positions. Circular dichroism spectroscopy, used to monitor thermal denaturations of a heterodimerizing leucine zipper system containing either valine (V) or asparagine (N) in the 'a' position, indicates that the V-N interaction is 2.3 kcal/mole less stable than an N-N interaction and 5.3 kcal/mole less stable than a V-V interaction. Thus, We propose that the presence of polar amino acids in novel positions of the 'a' position of Drosophila B-ZIP proteins has led to leucine zippers that homodimerize rather than heterodimerize. C1 Natl Lib Med, Comp Biol Branch, Natl Ctr Biotechnol Informat, Bethesda, MD 20814 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Univ Iowa, Dept Biol Sci, Iowa City, IA 52242 USA. RP Vinson, C (reprint author), Natl Lib Med, Comp Biol Branch, Natl Ctr Biotechnol Informat, Bethesda, MD 20814 USA. RI Landsman, David/C-5923-2009; OI Landsman, David/0000-0002-9819-6675 NR 60 TC 48 Z9 49 U1 0 U2 0 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD AUG PY 2002 VL 12 IS 8 BP 1190 EP 1200 DI 10.1101/gr.67902 PG 11 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 583WQ UT WOS:000177434300007 PM 12176927 ER PT J AU Thomas, JW Prasad, AB Summers, TJ Lee-Lin, SQ Maduro, VVB Idol, JR Ryan, JF Thomas, PJ McDowell, JC Green, ED AF Thomas, JW Prasad, AB Summers, TJ Lee-Lin, SQ Maduro, VVB Idol, JR Ryan, JF Thomas, PJ McDowell, JC Green, ED TI Parallel construction of orthologous sequence-ready clone contig maps in multiple species SO GENOME RESEARCH LA English DT Article ID ARTIFICIAL CHROMOSOME LIBRARY; HUMAN GENOME; MOUSE; REGIONS; HUMAN-CHROMOSOME-19; GENERATION; STRATEGIES; COVERAGE; MARKERS AB Comparison is a fundamental tool for analyzing DNA sequence. Interspecies sequence comparison is particularly powerful for inferring genome function and is based on the simple premise that conserved sequences are likely to be important. Thus, the comparison of a genomic sequence with its orthologous counterpart from another species is increasingly becoming an integral component of genome analysis. In ideal situations, such comparisons are performed with orthologous sequences from multiple species. To facilitate multispecies comparative sequence analysis, a robust and scalable strategy for simultaneously constructing sequence-ready bacterial artificial chromosome (BAC) contig maps from targeted genomic regions has been developed. Central to this approach is the generation and utilization of "universal" oligonucleotide-based hybridization probes ("overgo" probes), which are designed from sequences that are highly conserved between distantly related species. Large collections of these probes are used en masse to screen BAC libraries from multiple species in parallel, with the isolated clones assembled into physical contig maps. To validate the effectiveness of this strategy, efforts were focused on the construction of BAC-based physical maps from multiple mammalian species (chimpanzee, baboon, cat, dog, cow, and pig). Using available human and mouse genomic sequence and a newly developed computer program (soop) to design the requisite probes, sequence-ready maps were constructed in all species for a series of targeted regions totaling similar to16 Mb in the human genome. The described approach can be used to facilitate the multispecies comparative sequencing of targeted genomic regions and can be adapted for constructing BAC contig maps in other vertebrates. C1 NIH, Genome Technol Branch, NHGRI, Bethesda, MD 20892 USA. NIH, Intramural Sequencing Ctr, Bethesda, MD 20892 USA. RP Green, ED (reprint author), NIH, Genome Technol Branch, NHGRI, Bethesda, MD 20892 USA. EM egreen@nhgri.nih.gov RI Prasad, Arjun/C-6736-2008; OI Prasad, Arjun/0000-0002-1343-8664 NR 31 TC 57 Z9 61 U1 0 U2 3 PU COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT PI COLD SPRING HARBOR PA 1 BUNGTOWN RD, COLD SPRING HARBOR, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD AUG PY 2002 VL 12 IS 8 BP 1277 EP 1285 DI 10.1101/gr.283202 PG 9 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 583WQ UT WOS:000177434300015 PM 12176935 ER PT J AU Sultana, R Yu, CE Yu, J Munson, J Chen, DH Hua, WH Estes, A Cortes, F de la Barra, F Yu, DM Haider, ST Trask, BJ Green, ED Raskind, WH Disteche, CM Wijsman, E Dawson, G Storm, DR Schellenberg, GD Villacres, EC AF Sultana, R Yu, CE Yu, J Munson, J Chen, DH Hua, WH Estes, A Cortes, F de la Barra, F Yu, DM Haider, ST Trask, BJ Green, ED Raskind, WH Disteche, CM Wijsman, E Dawson, G Storm, DR Schellenberg, GD Villacres, EC TI Identification of a novel gene on chromosome 7q11.2 interrupted by a translocation breakpoint in a pair of autistic twins SO GENOMICS LA English DT Article ID QUANTITATIVE TRAIT; GENOMIC SCREEN; DISORDER; LINKAGE; LOCUS; 7Q AB We report here the identification and characterization of a novel gene (AUTS2) that spans the 7q11.2 break-point in a monozygotic twin pair concordant for autism and a t(7;20) (q11.2; p11.2) translocation. AUTS2 is 1.2 Mb and has 19 exons. The predicted protein is 1295 amino acids and does not correspond to any known protein. DNA sequence analysis of autism subjects and controls revealed 22 biallelic polymorphic sites. For all sites, both alleles were observed in both cases and controls. Thus no autism-specific mutation was observed. Association analysis with two exonic polymorphic sites and linkage analysis of four dinucleotide repeat markers, two within and two flanking AUTS2, was negative. Thus, although it is unlikely that AUTS2 is an autism susceptibility gene for idiopathic autism, it may be the gene responsible for the disorder in the twins studied here. C1 Univ Washington, Dept Psychiat & Behav Sci, Seattle, WA 98195 USA. Univ Washington, Dept Med, Seattle, WA 98195 USA. Univ Washington, Dept Mol Biotechnol, Seattle, WA 98195 USA. Univ Washington, Dept Pharmacol, Seattle, WA 98195 USA. Univ Washington, Dept Psychol, Seattle, WA 98195 USA. Univ Washington, Dept Biostat, Seattle, WA 98195 USA. Univ Washington, Dept Pathol, Seattle, WA 98195 USA. Univ Washington, Dept Neurol, Seattle, WA 98195 USA. Seattle Vet Affairs Med Ctr, Ctr Geriatr Res Educ & Clin, Seattle, WA 98195 USA. Univ Chile, Genet & Met Dis Unit, INTA, Santiago 13811, Chile. Univ Chile, Psychiat & Mental Hlth Dept, Santiago 13811, Chile. NHGRI, Genome Technol Branch, NIH, Bethesda, MD 20892 USA. RP Villacres, EC (reprint author), Univ Washington, Dept Psychiat & Behav Sci, Seattle, WA 98195 USA. RI 郑, 征/C-8514-2011; OI Wijsman, Ellen/0000-0002-2725-6669 FU NCRR NIH HHS [1P41 RR03655]; NICHD NIH HHS [P0I HD 34565, P50 HD33812] NR 20 TC 91 Z9 93 U1 4 U2 13 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD AUG PY 2002 VL 80 IS 2 BP 129 EP 134 DI 10.1006/geno.2002.6810 PG 6 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 583EK UT WOS:000177393500003 PM 12160723 ER PT J AU Ayyagari, R Demirci, FY Liu, JF Bingham, EL Stringham, H Kakuk, LE Boehnke, M Gorin, MB Richards, JE Sieving, PA AF Ayyagari, R Demirci, FY Liu, JF Bingham, EL Stringham, H Kakuk, LE Boehnke, M Gorin, MB Richards, JE Sieving, PA TI X-linked recessive atrophic macular degeneration from RPGR mutation SO GENOMICS LA English DT Article DE macular degeneration; RPGR; nonsense mutation; atrophic; splice site; RP3; CSNB1; NYX; X-linked ID RETINITIS-PIGMENTOSA RP3; CONE-ROD DYSTROPHY; EXON ORF15; GENE ABCR; FAMILIES; LOCALIZATION; LINKAGE; LOCUS AB We mapped a new X-linked recessive atrophic macular degeneration locus to Xp21.1-p11.4 and show allelic involvement of the gene RPGR, which normally causes severe peripheral retinal degeneration leading to global blindness. Ten affected males whom we examined had primarily macular atrophy causing progressive loss of visual acuity with minimal peripheral visual impairment. One additional male showed extensive macular degeneration plus peripheral loss of retinal pigment epithelium and choriocapillaries. Full-field electroretinograms (ERGs) showed normal cone and rod responses in some affected males despite advanced macular degeneration, emphasizing the dissociation of atrophic macular degeneration from generalized cone degenerations, including X-linked cone dystrophy (COD1). The RPGR gene nonsense mutation G-T at open reading frame (ORF)15+1164 cosegregated with the disease and may create a donor splice site. Identification of an RPGR mutation in atrophic macular degeneration expands the phenotypic range associated with this gene and provides a new tool for the dissection of the relationship between clinically different retinal pathologies. C1 Univ Michigan, Dept Ophthalmol & Visual Sci, Ann Arbor, MI 48105 USA. Univ Pittsburgh, Dept Ophthalmol, Pittsburgh, PA 15213 USA. Univ Michigan, Dept Biostat, Ann Arbor, MI 48109 USA. NEI, NIH, Bethesda, MD 20892 USA. RP Sieving, PA (reprint author), Univ Michigan, Dept Ophthalmol & Visual Sci, Ann Arbor, MI 48105 USA. OI Demirci, F. Yesim/0000-0001-6907-9843 FU NEI NIH HHS [EY13130, EY07003, EY13198] NR 33 TC 74 Z9 76 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD AUG PY 2002 VL 80 IS 2 BP 166 EP 171 DI 10.1006/geno.2002.6815 PG 6 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 583EK UT WOS:000177393500010 PM 12160730 ER PT J AU Anisimov, SV Tarasov, KV Stern, MD Lakatta, EG Boheler, KR AF Anisimov, SV Tarasov, KV Stern, MD Lakatta, EG Boheler, KR TI A quantitative and validated SAGE transcriptome reference for adult mouse heart SO GENOMICS LA English DT Article DE heart; mouse; SAGE; gene expression array; transcriptome; gene profile ID ABUNDANCE MESSENGER-RNAS; GENE-EXPRESSION; SERIAL ANALYSIS; MYOCARDIAL-INFARCTION; SKELETAL-MUSCLE; CDNA MICROARRAY; GENOME; IDENTIFICATION; SEQUENCE; CELLS AB Transcriptome comparisons facilitate the identification of developmental, aging, and disease-related genes. To quantify the functionally active genome of adult C57BL/6 mouse heart (AMH), we used serial analysis of gene expression (SAGE) to sequence a total of 88,860 tags or 23,941 unique tags. Over 66% of the unique tags matched either known genes or ESTs. Mitochondrial transcripts accounted for 18.7% of the total transcripts, whereas sarcomeric proteins accounted for 3.2% of all tags. After comparison of AMH expression profiles obtained by SAGE and cDNA arrays, we observed numerous quantitative discrepancies (for example: arrays, mt-Col > mt-Co2 > mt-Co3; SAGE, mt-Col >> mt-Co3 : mt-Co2). We carried out quantitative PCR analyses as an independent test of transcript abundance and determined that SAGE yielded quantitatively reliable data. These SAGE results thus represent the first quantitative expression profile of AMH and serve as a reliable transcriptome reference to identify dynamic changes in cardiac gene expression. C1 NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Boheler, KR (reprint author), NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. EM bohelerk@grc.nia.nih.gov NR 38 TC 29 Z9 31 U1 0 U2 2 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD AUG PY 2002 VL 80 IS 2 BP 213 EP 222 DI 10.1006/geno.2002.6821 PG 10 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 583EK UT WOS:000177393500015 PM 12160735 ER PT J AU Kosary, C Trimble, EL AF Kosary, C Trimble, EL TI Treatment and survival for women with fallopian tube carcinoma: A population-based study SO GYNECOLOGIC ONCOLOGY LA English DT Article ID CANCER AB Objective. The objective of this study was to evaluate treatment and survival for women with fallopian tube carcinoma in a population-based data set. Methods. Using the National Cancer Institute's Surveillance, Epidemiology, and End Results program, we identified 416 women with fallopian tube carcinoma diagnosed between 1990 and 1997. We analyzed treatment and 5-year relative survival. We also compared survival to that of 9032 women with epithelial ovarian cancer diagnosed between 1991 and 1997. Results. Almost half of those diagnosed with stage I/II disease did not undergo surgical evaluation of lymph nodes. Most women with stage I/II disease were treated with surgery alone, while most women with stage III/IV disease were treated with surgery and chemotherapy. Five-year relative survival by FIGO stage was as follows: stage I (N=102), 95%; stage II (N=29), 75%; stage III (N=52), 69%; stage IV (N=151), 45%. Conclusions. We observed better survival, stage by stage, for women with fallopian tube carcinoma than for women with epithelial ovarian cancer in this population-based data set. It is possible that some patients with advanced, bulky carcinoma arising in the fallopian tube may have been classified as having ovarian or primary peritoneal cancer. Women with fallopian tube cancer should be treated in accordance with the same guidelines for surgical staging, debulking, and adjuvant chemotherapy as for women with epithelial ovarian cancer. Further studies, both laboratory and clinical, are needed to delineate the differences between fallopian and ovarian cancers. C1 Natl Canc Inst, Bethesda, MD 20892 USA. RP Kosary, C (reprint author), Natl Canc Inst, Bethesda, MD 20892 USA. NR 7 TC 43 Z9 45 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0090-8258 J9 GYNECOL ONCOL JI Gynecol. Oncol. PD AUG PY 2002 VL 86 IS 2 BP 190 EP 191 DI 10.1006/gyno.2002.6743 PG 2 WC Oncology; Obstetrics & Gynecology SC Oncology; Obstetrics & Gynecology GA 581AB UT WOS:000177268000015 PM 12144827 ER PT J AU Steck, DJ Alavanja, MCR Field, RW Parkhurst, MA Bates, DJ Mahaffey, JA AF Steck, DJ Alavanja, MCR Field, RW Parkhurst, MA Bates, DJ Mahaffey, JA TI Po-210 implanted in glass surfaces by long term exposure to indoor radon SO HEALTH PHYSICS LA English DT Article DE radon progeny; exposure, radiation; epidemiology; monitoring, air ID LUNG-CANCER; RETROSPECTIVE ASSESSMENT; RN-222 EXPOSURE; RISK; DWELLINGS; DETECTORS; MODEL AB Recent epidemiologic investigations of the relationship between residential radon gas exposure and lung cancer relied on contemporary radon gas measurements to estimate past radon gas exposures. Significant uncertainties in these exposure estimates can arise from year-to-year variation of indoor radon concentrations and subject mobility. Surface implanted Po-210 has shown potential for improving retrospective radon gas exposure estimates. However, in previous studies, the ability of implanted Po-210 activity to reconstruct cumulative radon gas exposure was not tested because glass was not available from homes with known radon-gas concentration histories. In this study, we tested the validity of the retrospective radon gas reconstruction using implanted Po-210 surface activity by measuring glass surfaces from homes whose annual-average radon gas concentrations had been measured almost every year during two decades. Regression analysis showed a higher correlation between measured surface activity and cumulative radon gas exposure in these homes (R-2>0.8) than was observed in homes where only contemporary radon gas measurements were available. The regression slope (0.57 ky m(-1)) was consistent with our earlier retrospective results. Surface activity measurements were as reliable for retrospective radon gas exposure reconstruction as yearlong gas measurements. Both methods produced estimates that were within 25% of the long-term average radon gas concentrations in a home. Surface measurements can be used for home screening tests because they can provide rapid, reliable estimates of past radon gas concentrations. Implanted Po-210 measurements are also useful in retrospective epidemiologic studies that include participants who may have been exposed to highly variable radon concentrations in previously occupied or structurally modified homes. C1 St Johns Univ, Dept Phys, Collegeville, MN 56321 USA. NCI, Bethesda, MD 20814 USA. Univ Iowa, Coll Publ Hlth, Dept Epidemiol, Iowa City, IA 52242 USA. Pacific NW Natl Lab, Richland, WA 99352 USA. RP Steck, DJ (reprint author), St Johns Univ, Dept Phys, Collegeville, MN 56321 USA. EM dsteck@csbsju.edu FU NCI NIH HHS [R01 CA859422] NR 35 TC 20 Z9 22 U1 2 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0017-9078 EI 1538-5159 J9 HEALTH PHYS JI Health Phys. PD AUG PY 2002 VL 83 IS 2 BP 261 EP 271 DI 10.1097/00004032-200208000-00011 PG 11 WC Environmental Sciences; Public, Environmental & Occupational Health; Nuclear Science & Technology; Radiology, Nuclear Medicine & Medical Imaging SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Nuclear Science & Technology; Radiology, Nuclear Medicine & Medical Imaging GA 574JR UT WOS:000176886200011 PM 12132714 ER PT J AU Legler, J Breen, N Meissner, H Malec, D Coyne, C AF Legler, J Breen, N Meissner, H Malec, D Coyne, C TI Predicting patterns of mammography use: A geographic perspective on national needs for intervention research SO HEALTH SERVICES RESEARCH LA English DT Article DE Bayesian hierarchical modeling; mammography; intervention research; geography ID HEALTH-INTERVIEW-SURVEY; SCREENING MAMMOGRAPHY; UNITED-STATES; BREAST; CANCER; MORTALITY; INCOME; WOMEN; RACE AB Objective. To introduce a methodology for planning preventive health service research that takes into account geographic context. Data Sources. National Health Inter-view Survey (NHIS) self-reports of mammography within the past two years, 1987, and 1993-94. Area Resource File (ARF), 1990. Database of mammography intervention research studies conducted from 1984 to 1994. Design. Bayesian hierarchical modeling describes mammography as a function of county-level socioeconomic data and explicitly estimates the geographic variation unexplained by the county-level data. This model produces county use estimates (both NHIS-sampled and unsampled), which are aggregated for entire states. The locations of intervention research studies are examined in light of the statewide mammography utilization estimates. Data Extraction. Individual level NHIS data were merged with county-level data from the ARF. Principal Findings. State maps reveal the estimated distribution of mammography utilization and intervention research. Eighteen states with low mammography use reported no intervention research activity. County-level occupation and education were important predictors for younger women in 1993-94. In 1987, they were not predictive for any demographic group. Conclusions. Opportunities exist to improve the planning of future intervention research by considering geographic context. Modeling results suggest that the choice of predictors be tailored to both the population and the time period under study when planning interventions. C1 NCI, Stat Res & Applicat Branch, Surveillance Res Program, Bethesda, MD 20892 USA. NCI, Appl Res Program, Hlth Serv & Econ Branch, Bethesda, MD USA. NCI, Behav Res Program, Appl Canc Screening Res Branch, Bethesda, MD 20892 USA. US Bur Census, Div Stat Res, Washington, DC USA. W Virginia Univ, Morgantown, WV USA. RP Legler, J (reprint author), NCI, Stat Res & Applicat Branch, Surveillance Res Program, EPN Room 4094,6130 Execut Blvd,MSC 7359, Bethesda, MD 20892 USA. NR 33 TC 17 Z9 17 U1 4 U2 5 PU BLACKWELL PUBL LTD PI OXFORD PA 108 COWLEY RD, OXFORD OX4 1JF, OXON, ENGLAND SN 0017-9124 J9 HEALTH SERV RES JI Health Serv. Res. PD AUG PY 2002 VL 37 IS 4 BP 929 EP 947 DI 10.1034/j.1600-0560.2002.59.x PG 19 WC Health Care Sciences & Services; Health Policy & Services SC Health Care Sciences & Services GA 591PQ UT WOS:000177889800008 PM 12236391 ER PT J AU Wu, T Han, C Lunz, JG Michalopoulos, G Shelhamer, JH Demetris, AJ AF Wu, T Han, C Lunz, JG Michalopoulos, G Shelhamer, JH Demetris, AJ TI Involvement of 85-kd cytosolic phospholipase A(2) and cyclooxygenase-2 in the proliferation of human cholangiocarcinoma cells SO HEPATOLOGY LA English DT Article ID HEPATOCYTE GROWTH-FACTOR; BILIARY EPITHELIAL-CELLS; HUMAN INTRAHEPATIC CHOLANGIOCARCINOMA; HUMAN HEPATOCELLULAR-CARCINOMA; ACTIVATED PROTEIN-KINASE; IN-VITRO; FACTOR/SCATTER FACTOR; EXPRESSION; INTERLEUKIN-6; COX-2 AB Cyclooxygenase 2 (COX-2) and cytosolic phospholipase A(2) (cPLA(2)) are the crucial rate-limiting enzymes in prostaglandin (PG) metabolism that show increased expression in a number of human cancers, including cholangiocarcinomas; and treatment of cholangiocarcinoma cell lines with COX-2 inhibitors can decrease proliferation. Cholangiocarcinomas also produce and proliferate in response to nonneoplastic biliary epithelial cell mitogens, such as interleukin 6 (IL-6) and hepatocyte growth factor (HGF). This study was designed to determine whether there is any relationship between eicosanoid metabolism and growth stimulation by IL-6 and HGF, two important biliary epithelial cell and cholangiocarcinoma mitogens. Incubation of SG231,a well-characterized human cholangiocarcinoma cell line, with HGF, IL-6, PGE(2), or PGF(2)alpha resulted in significantly increased cell growth. HGF and IL-6 also induced a rapid release of arachidonic acid (AA) from SG231 and increased the synthesis of PGE(2) and PGF(2)alpha. The cPLA2 inhibitor arachidonyl fluorophosphonate (MAFP) and the COX-2 inhibitor NS-398 significantly inhibited HGF- and IL-6-induced release of AA, PG synthesis, and proliferation in SG231 cells as well as two other human cholangiocarcinoma cell lines, HuCCT1 and CC-LP-1 cells. Thus, PGs alone can induce cholangiocarcinoma growth, and the HGF- and IL-6-induced proliferation is mediated, at least in part, by PGs. HGF and IL-6 also induced a rapid phosphorylation of cPLA2 (within 1 minute) but did not alter cPLA2 and COX-2 protein expression. The HGF- and IL-6-induced cPLA2 phosphorylation was blocked by the inhibitors of p38 and p42/44 MAP kinases, protein kinase C, calmodulin kinase, and tyrosine kinase, showing that HGF- and IL-6-induced AA release and PG production are mediated by phosphorylation of cPLA2. In conclusion, molecular pathways link classic biliary epithelial cell mitogens to PG metabolism constituents in cholangiocarcinoma growth, which may be exploited as potential therapeutic targets. C1 Univ Pittsburgh, Dept Pathol, Sch Med, Pittsburgh, PA 15213 USA. NIH, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Wu, T (reprint author), Univ Pittsburgh, Presbyterian Hosp, Sch Med, Dept Pathol, C902,200 Lothrop St, Pittsburgh, PA 15213 USA. FU NIDDK NIH HHS [R01 DK49615] NR 50 TC 53 Z9 56 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD AUG PY 2002 VL 36 IS 2 BP 363 EP 373 DI 10.1053/jhep.2002.34743 PG 11 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 577WG UT WOS:000177085100012 PM 12143044 ER PT J AU Roix, J Misteli, T AF Roix, J Misteli, T TI Genomes, proteomes, and dynamic networks in the cell nucleus SO HISTOCHEMISTRY AND CELL BIOLOGY LA English DT Article; Proceedings Paper CT 44th Symposium of the Society-for-Histochemistry CY SEP 25-28, 2002 CL VLISSINGEN, NETHERLANDS SP Soc Histochem DE genome; proteome; network; nucleus ID LIVING HUMAN-CELLS; IN-VIVO ANALYSIS; INTERPHASE NUCLEI; RAPID EXCHANGE; CHROMOSOME TERRITORIES; TRANSCRIPTOME MAP; MAMMALIAN-CELLS; GENE-EXPRESSION; SPLICING FACTOR; COILED BODIES AB Post-genome era experimental strategies seek to understand cellular pathways globally rather than through examination of individual components. Genomics and proteomics provide the experimental tools to establish the framework of gene and protein pathways present in a cell. These methods are complemented by emerging in vivo microscopy approaches, which permit placement of pathways within the architectural context of the cell. Analysis of dynamic live cell microscopy data combined with computational analyses finally allows the quantitative, mechanistic description of protein properties and pathways operating in living cells. Here we discuss how genomics and proteomics are changing the study of the cell nucleus and how in vivo microscopy methods have contributed to our changing conceptual and mechanistic understanding of nuclear architecture and function. Mapping of gene loci and genome regions are beginning to reveal organizational principles of the genome within the cell nucleus, proteomic analysis of subnuclear compartments and the gene expression machinery is providing insights into the molecular nature of nuclear events, and in vivo microscopy is illuminating the dynamic nature of nuclear organization. Initial findings from these efforts make clear that nuclear and cellular behavior can not be described by linear pathways. New tools such as computational modeling are providing evidence that emerging concepts such as network organizations and stochastic interactions are crucially important for cellular function and organization. These insights are changing our view of the nucleus and the cell as a whole. C1 NCI, NIH, Bethesda, MD 20892 USA. RP Misteli, T (reprint author), NCI, NIH, Bethesda, MD 20892 USA. EM mistelit@mail.nih.gov NR 79 TC 25 Z9 25 U1 0 U2 0 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0948-6143 J9 HISTOCHEM CELL BIOL JI Histochem. Cell Biol. PD AUG PY 2002 VL 118 IS 2 BP 105 EP 116 DI 10.1007/s00418-002-0446-7 PG 12 WC Cell Biology; Microscopy SC Cell Biology; Microscopy GA 592LH UT WOS:000177938800003 PM 12189514 ER PT J AU Mononen, N Ikonen, T Autio, V Rokman, A Matikainen, MP Tammela, TLJ Kallioniemi, OP Koivisto, PA Schleutker, J AF Mononen, N Ikonen, T Autio, V Rokman, A Matikainen, MP Tammela, TLJ Kallioniemi, OP Koivisto, PA Schleutker, J TI Androgen receptor CAG polymorphism and prostate cancer risk SO HUMAN GENETICS LA English DT Article ID BULBAR MUSCULAR-ATROPHY; VITAMIN-D-RECEPTOR; LINKAGE DISEQUILIBRIUM; REPEAT LENGTH; GENE; HYPERPLASIA; MEN; ASSOCIATION; EXPRESSION; POPULATION AB Recent studies have suggested that polymorphisms of the androgen receptor gene (AR) may influence the risk of prostate cancer (PC) development and progression. Here, we analyzed the length of the CAG repeat of the AR gene in 1363 individuals, including patients with PC, benign prostate hyperplasia (BPH), and population controls. There was a tendency for short CAG repeats to be associated with PC. The Odds Ratio (OR) for PC was 1.47 (P=0.05) when individuals with short CAG repeats (less than or equal to18) were compared with those having long repeats (>18). CAG repeat length was not significantly associated with family history, disease stage, grade, age at diagnosis, prostate-specific antigen (PSA) level at diagnosis, or prognosis of the patients. Unexpectedly, short CAG repeats were significantly less common in patients with BPH compared with controls (OR=0.47, P=0.03). Our results suggest that the CAG polymorphism of the AR gene is unlikely to have a major role in the development or progression of PC in the Finnish population. The association of CAG repeats with the risk of BPH warrants further study. C1 Univ Tampere, Canc Genet Lab, Inst Med Technol, Tampere 33520, Finland. Tampere Univ Hosp, Canc Genet Lab, Inst Med Technol, Tampere 33520, Finland. Tampere Univ Hosp, Dept Clin Genet, Tampere 33521, Finland. Univ Tampere, Tampere Sch Publ Hlth, Tampere 33014, Finland. Tampere Univ Hosp, Dept Urol, Tampere 33521, Finland. Univ Tampere, Sch Med, Tampere 33521, Finland. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. RP Mononen, N (reprint author), Univ Tampere, Canc Genet Lab, Inst Med Technol, Lenkkeilijankatu 6, Tampere 33520, Finland. RI Kallioniemi, Olli/H-5111-2011; Kallioniemi, Olli/H-4738-2012 OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332 FU NHGRI NIH HHS [N01-HG-55389] NR 27 TC 57 Z9 59 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD AUG PY 2002 VL 111 IS 2 BP 166 EP 171 DI 10.1007/s00439-002-0776-5 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 591PV UT WOS:000177890200007 PM 12189490 ER PT J AU Ben-Zeev, B Levy-Nissenbaum, E Lahat, H Anikster, Y Shinar, Y Brand, N Gross-Tzur, V MacGregor, D Sidi, R Kleta, R Frydman, M Pras, E AF Ben-Zeev, B Levy-Nissenbaum, E Lahat, H Anikster, Y Shinar, Y Brand, N Gross-Tzur, V MacGregor, D Sidi, R Kleta, R Frydman, M Pras, E TI Megalencephalic leukoencephalopathy with subcortical cysts; a founder effect in Israeli patients and a higher than expected carrier rate among Libyan Jews SO HUMAN GENETICS LA English DT Article ID MILD CLINICAL COURSE; LEUKODYSTROPHY AB Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a progressive inherited neurological disorder characterized by macrocephaly, deterioration in motor functions and cerebellar ataxia. In Israel the disease is found in an increased frequency among Libyan Jews. The disease is caused by mutations in the MLC1 gene, which encodes a putative CNS membrane transporter. We describe three novel mutations (p.G59E, p.P92S, and 134-136insC) in seven MLC families. One of these mutations, p.G59E, was found in the vast majority of MLC patients in Israel. Screening of 200 normal Libyan Jewish individuals for the p.G59E mutation, revealed a carrier rate of 1/40 compared with an expected carrier rate of 1/81. Several explanations could account for this difference the most likely one is an admixture of the Libyan Jewish population. C1 Chaim Sheba Med Ctr, Danek Gartner Inst Human Genet, IL-52621 Tel Hashomer, Israel. Chaim Sheba Med Ctr, Pediat Neurol Unit, IL-52621 Tel Hashomer, Israel. NICHHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. Chaim Sheba Med Ctr, Heller Inst Med Sci, IL-52621 Tel Hashomer, Israel. Shaare Zedek Med Ctr, Pediat Neurol Unit, IL-81031 Jerusalem, Israel. Hosp Sick Children, Div Neurol, Toronto, ON M5G 1X8, Canada. Tel Aviv Univ, Sackler Sch Med, IL-69978 Tel Aviv, Israel. RP Pras, E (reprint author), Chaim Sheba Med Ctr, Danek Gartner Inst Human Genet, IL-52621 Tel Hashomer, Israel. NR 14 TC 18 Z9 19 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD AUG PY 2002 VL 111 IS 2 BP 214 EP 218 DI 10.1007/s00439-002-0770-y PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 591PV UT WOS:000177890200013 PM 12189496 ER PT J AU Jensen, JT Schwinof, KM Zelinski-Wooten, MB Conti, M DePaolo, LV Stouffer, RL AF Jensen, JT Schwinof, KM Zelinski-Wooten, MB Conti, M DePaolo, LV Stouffer, RL TI Phosphodiesterase 3 inhibitors selectively block the spontaneous resumption of meiosis by macaque oocytes in vitro SO HUMAN REPRODUCTION LA English DT Article DE contraception; meiosis; non-human primate; oocyte; phosphodiesterase 3 inhibitor ID IN-VITRO; MOUSE OOCYTE; CAMP LEVELS; SPONTANEOUS MATURATION; NONHUMAN-PRIMATES; MONKEY OOCYTES; RHESUS-MONKEYS; FERTILIZATION; INVITRO; CELLS AB BACKGROUND: The purpose of this study was to determine whether phosphodiesterase (PDE) 3 inhibitors selectively prevent the resumption of meiosis in primates. METHODS: Immature oocytes (intact germinal vesicles) obtained from large pre-ovulatory follicles following ovarian stimulation in rhesus macaques were incubated with or without various doses of the PDE3 inhibitors, Cilostamide, Milrinone or ORG 9935, or a selective PDE4 inhibitor, Rolipram. Oocytes were observed for germinal vesicle breakdown (GVBD) as an indicator of resumption of meiosis. RESULTS: At 24 h, 72 of 121 (60%) control oocytes progressed to GVBD compared with 9/34 (27%, P < 0.01), 4/36 (11.1%, P < 0.01) and 0/28 (0%, P < 0.01) oocytes incubated with ORG 9935 at 0.1, 0.5 and 1.0 μmol/l respectively. Similar results were achieved at 24 h with 1.0 μmol/l Cilostamide (2/24 oocytes, 8%, P < 0.01) and 100 mumol/l Milrinone (2/32, 6%, P < 0.01). In contrast, no significant difference in GVBD was noted between control oocytes and those incubated with up to 100 μmol/l Rolipram for 24 h (43/58, 74%) or 48 h (44/58, 76%). CONCLUSIONS: These experiments establish the specificity and dose-dependent ability of PDE3, but not PDE4, inhibitors to block resumption of meiosis in macaque oocytes in vitro. Thus, PDE3 inhibitors have potential use as contraceptives in primates. C1 Oregon Hlth Sci Univ, Dept Obstet & Gynecol, Oregon Reg Primate Res Ctr, Portland, OR 97201 USA. Oregon Hlth Sci Univ, Oregon Reg Primate Res Ctr, Div Reprod Sci, Portland, OR 97201 USA. Oregon Hlth Sci Univ, Oregon Reg Primate Res Ctr, Div Reprod Sci, Beaverton, OR 97006 USA. Oregon Hlth Sci Univ, Oregon Reg Primate Res Ctr, Dept Physiol & Pharmacol, Beaverton, OR 97006 USA. Stanford Univ, Sch Med, Dept Obstet & Gynecol, Div Reprod Biol, Stanford, CA 94305 USA. NICHD, Reprod Sci Branch, Populat Res Ctr, NIH, Bethesda, MD USA. RP Jensen, JT (reprint author), Oregon Hlth Sci Univ, Dept Obstet & Gynecol, Oregon Reg Primate Res Ctr, Portland, OR 97201 USA. OI Jensen, Jeffrey/0000-0002-4733-1224 FU NCRR NIH HHS [RR00163]; NICHD NIH HHS [HD 01243-03, U54 HD 031398, U54 HD 18185] NR 36 TC 42 Z9 45 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0268-1161 J9 HUM REPROD JI Hum. Reprod. PD AUG PY 2002 VL 17 IS 8 BP 2079 EP 2084 DI 10.1093/humrep/17.8.2079 PG 6 WC Obstetrics & Gynecology; Reproductive Biology SC Obstetrics & Gynecology; Reproductive Biology GA 580HD UT WOS:000177228400027 PM 12151440 ER PT J AU Bakalov, VK Vanderhoof, VH Bondy, CA Nelson, LM AF Bakalov, VK Vanderhoof, VH Bondy, CA Nelson, LM TI Adrenal antibodies detect asymptomatic auto-immune adrenal insufficiency in young women with spontaneous premature ovarian failure SO HUMAN REPRODUCTION LA English DT Article DE Addison disease; adrenal antibodies; adrenal insufficiency; 21-hydroxylase antibodies; POF ID ADDISONS-DISEASE; AUTOIMMUNITY; AUTOANTIBODIES; CORTEX AB BACKGROUND: Auto-immune adrenal insufficiency is a potentially fatal disorder. Young women with spontaneous premature ovarian failure (POF) are at increased risk of developing this condition. METHODS: We further characterized auto-immune adrenal insufficiency in this population by conducting an in-depth cross-sectional evaluation of adrenal function in a series of 123 women. RESULTS: We uncovered a new diagnosis of adrenal insufficiency in four women [3.2%, 95% confidence interval (CI) 0.2-6.4%]. All four tested positive for adrenal antibodies as detected by a clinically available indirect immunofluorescence assay. A positive adrenal antibody test was highly associated with adrenal insufficiency while a negative test was associated with normal adrenal function in all cases (P < 0.001). Adrenal antibodies increased the pretest probability of adrenal insufficiency from 3.2 to 67%. As a screening method the cortisol response during a standard adrenocorticotrophic hormone (ACTH) stimulation test gave two false positive results (1.7%, upper 95% confidence limit 5.0%). CONCLUSIONS: Our findings suggest that measuring adrenal antibodies would be an effective screening method by which to detect auto-immune adrenal insufficiency in young women with spontaneous POF. The standard ACTH stimulation test should be reserved to confirm adrenal insufficiency in women with adrenal antibodies, or those with signs and symptoms of adrenal insufficiency. C1 NICHHD, Sect Womens Hlth Res, Dev Endocrinol Branch, NIH, Bethesda, MD 20829 USA. RP Nelson, LM (reprint author), NICHHD, Sect Womens Hlth Res, Dev Endocrinol Branch, NIH, Bldg 10,Room 10N262, Bethesda, MD 20829 USA. NR 24 TC 44 Z9 49 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0268-1161 J9 HUM REPROD JI Hum. Reprod. PD AUG PY 2002 VL 17 IS 8 BP 2096 EP 2100 DI 10.1093/humrep/17.8.2096 PG 5 WC Obstetrics & Gynecology; Reproductive Biology SC Obstetrics & Gynecology; Reproductive Biology GA 580HD UT WOS:000177228400030 PM 12151443 ER PT J AU Malyguine, A Derby, E Brooks, A Reddy, V Baseler, M Sayers, T AF Malyguine, A Derby, E Brooks, A Reddy, V Baseler, M Sayers, T TI Study of diverse mechanisms of cell-mediated cytotoxicity in gene-targeted mice using flow cytometric cytotoxicity assay SO IMMUNOLOGY LETTERS LA English DT Article DE cell-mediated cytotoxicity; flow cytometry; gene-targeted mice; perforin; Fas ID APOPTOSIS AB Cytotoxic lymphocytes kill tumor or virus-infected target cells utilizing two mechanisms-(1) release of lytic granules (containing perforin and granzymes), and (2) Fits ligand (FasL)/Fas or TIN-initiated apoptosis. We have examined mechanisms of target cell lysis by effector T cells from gene-targeted and mutant mice using a new Flow Cytometric Cytotoxicity Assay (FC Assay). Target cells were labeled with PKH67 dye. Cell death was estimated by 7-AAD inclusion and annexin V-PE binding. A direct correlation has been found between the percentage of dead target cells in FC Assay and the results of In-111 release cytotoxicity assay when effector T cells from either Pfp -/- (perform knockout) or gld (non-functional Fas Ligand) mice were used. As shown by the 4 h FC assay, the granule-mediated mechanism was utilized by T cells from gld mice. In contrast, T cells from Pfp -/- mice used death receptor-mediated lysis. Therefore. cytotoxic cells from gene-targeted and mutant mice can serve as valuable tools for studying different mechanisms of cell-mediated cytotoxicity, and the FC assay could be applied irrespective of which cytotoxic effector pathway is involved. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick Inc, Frederick, MD 21702 USA. RP Malyguine, A (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick Inc, POB B, Frederick, MD 21702 USA. RI Sayers, Thomas/G-4859-2015 FU NCI NIH HHS [N01 CO 56000] NR 8 TC 10 Z9 10 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-2478 J9 IMMUNOL LETT JI Immunol. Lett. PD AUG 1 PY 2002 VL 83 IS 1 BP 55 EP 59 AR PII S0165-2478(02)00048-2 DI 10.1016/S0165-2478(02)00048-2 PG 5 WC Immunology SC Immunology GA 576KT UT WOS:000177004200007 PM 12057855 ER PT J AU McDowell, MA Marovich, M Lira, R Braun, M Sacks, D AF McDowell, MA Marovich, M Lira, R Braun, M Sacks, D TI Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent SO INFECTION AND IMMUNITY LA English DT Article ID EXPERIMENTAL CUTANEOUS LEISHMANIASIS; LEVEL IL-12 PRODUCTION; INDIAN KALA-AZAR; T-HELPER CELLS; INTERFERON-GAMMA; IN-VIVO; PROTECTIVE IMMUNITY; MAJOR PROMASTIGOTES; MONONUCLEAR-CELLS; SUSCEPTIBLE MICE AB A major question in the study of leishmaniasis is what dictates clinical disease expression produced by different Leishmania species, i.e., cutaneous versus systemic and healing versus northealing. Animal models using a Leishmania species associated with self-limiting cutaneous disease (L. major) have revealed that protective immunity requires CD40/CD40 ligand (CD40L)-dependent, interleukin-12 (IL-12)-driven Th1 responses. We recently showed that L. major can prime human dendritic cells (DCs) for CD40L-triggered IL-12p70 secretion and that these cells can drive a Th1 response in autologous T cells from sensitized individuals. Here we show that in contrast to L. major, Leishmania species responsible for visceral disease (L. donovani), as well as species associated with persistent, cutaneous lesions and occasional systemic disease (L. tropica), did not induce CD40L-dependent IL-12p70 production, despite comparable levels of uptake by DCs. Up-regulated surface expression of CD40 did not correlate with IL-12p70 production, and appreciable CD40L-induced IL-12p40 secretion was observed in uninfected as well as infected DCs, regardless of species. Reverse transcription-PCR analysis confirmed that the production of heterodimeric IL-12 was limited by expression of IL-12p35 mRNA, which was dependent on both a microbial priming signal and CD40 engagement for its high-level induction. The intrinsic differences in the ability of Leishmania species to prime DCs for CD40L-dependent IL-12p70 secretion may account, at least in part, for the evolution of healing and northealing forms of leishmanial disease. C1 NIAID, Parasit Dis Lab, Ctr Dr MSC 0425, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Sacks, D (reprint author), NIAID, Parasit Dis Lab, Ctr Dr MSC 0425, NIH, Bldg 4,Rm 126, Bethesda, MD 20892 USA. NR 57 TC 64 Z9 67 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 3994 EP 4001 DI 10.1128/IAI.70.8.3994-4001.2002 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400002 PM 12117904 ER PT J AU Rogers, WO Weiss, WR Kumar, A Aguiar, JC Tine, JA Gwadz, R Harre, JG Gowda, K Rathore, D Kumar, S Hoffman, SL AF Rogers, WO Weiss, WR Kumar, A Aguiar, JC Tine, JA Gwadz, R Harre, JG Gowda, K Rathore, D Kumar, S Hoffman, SL TI Protection of rhesus Macaques against lethal plasmodium knowlesi malaria by a heterologous DNA priming and poxvirus boosting immunization regimen SO INFECTION AND IMMUNITY LA English DT Article ID VACCINIA VIRUS; DENDRITIC CELLS; FALCIPARUM-MALARIA; PLASMID DNA; ANTIGEN; EXPRESSION; INDUCTION; IMMUNOGENICITY; IMMUNITY; ANTIBODY AB We tested a cytokine-enhanced, multiantigen, DNA priming and poxvirus boosting vaccine regimen for prevention of malaria in the Plasmodium knowlesi-rhesus macaque model system. Animals were primed with a mixture of DNA plasmids encoding two preerythrocytic-stage proteins and two erythrocytic-stage proteins from P. knowlesi and combinations of the cytokines granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor necrosis factor alpha and were boosted with a mixture of four recombinant, attenuated vaccinia virus strains encoding the four P. knowlesi antigens. Two weeks after boosting, the geometric mean immunofluorescence titers in the immunized groups against sporozoites and infected erythrocytes ranged from 160 to 8,096 and from 1,810 to 5,120, respectively. The geometric mean anti-P. knowlesi circumsporozoite protein (PkCSP) titers ranged from 1,761 to 24,242. Peripheral blood mononuclear cells (PBMC) from the immunized monkeys produced gamma interferon (IFN-gamma) in response to incubation with pooled peptides from the PkCSP at frequencies of 10 to 571 spot-forming cells/10(6) PBMC. Following challenge with 100 infectious P. knowlesi sporozoites, 2 of 11 immunized monkeys were sterilely protected, and 7 of the 9 infected monkeys resolved their parasitemias spontaneously. In contrast, all four controls became infected and required treatment for overwhelming parasitemia. Early protection was strongly associated with IFN-gamma responses against a pool of peptides from the preerythrocytic-stage antigen, PkCSP. These findings demonstrate that a multistage, multiantigen, DNA priming and poxvirus boosting vaccine regimen can protect nonhuman primates from an otherwise lethal malaria sporozoite challenge. C1 Walter Reed Army Inst Res, Div Vet Med, Silver Spring, MD 20910 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20889 USA. Virogenet Corp, Troy, NY 12180 USA. USN, Res Ctr, Malaria Program, Silver Spring, MD 20910 USA. RP Rogers, WO (reprint author), US Dept State, Naval Med Res Unit 3, 2020 Accra Pl, Washington, DC 20521 USA. NR 31 TC 60 Z9 61 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 4329 EP 4335 DI 10.1128/IAI.70.8.4329-4335.2002 PG 7 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400040 PM 12117942 ER PT J AU Xu, DQ Cisar, JO Ambulos, N Burr, DH Kopecko, DJ AF Xu, DQ Cisar, JO Ambulos, N Burr, DH Kopecko, DJ TI Molecular cloning and characterization of genes for Shigella sonnei form IO polysaccharide: Proposed biosynthetic pathway and stable expression in a live Salmonella vaccine vector SO INFECTION AND IMMUNITY LA English DT Article ID O-ANTIGEN GENES; ESCHERICHIA-COLI; PSEUDOMONAS-AERUGINOSA; BIVALENT VACCINE; TYPHOID VACCINE; DNA-SEQUENCE; PLASMID; LIPOPOLYSACCHARIDE; STRAIN; CONSTRUCTION AB The gene region for biosynthesis of Shigella sonnei form I O polysaccharide (O-Ps) and flanking sequences, totaling >18 kb, was characterized by deletion analysis to define a minimal construct for development of Salmonella-based live vaccine vector strains. Lipopolysaccharide (LPS) expression and DNA sequence studies of plasmid deletion derivatives indicated form I O-Ps expression from a 12.3-kb region containing a putative promoter and 10 contiguous open reading frames (ORFs), one of which is the transposase of IS630. A detailed binsynthetic pathway, consistent with the predicted functions of eight of the nine essential ORFs and the form I O-Ps structure, is proposed. Further sequencing identified partial IS elements (i.e., IS91 and IS630) and wzz upstream of the form I coding region and a fragment of aqpZ and additional full or partial IS elements (i.e., IS629, IS91, and IS911) downstream of this region. The stability of plasmid-based form I O-Ps expression was greater from low-copy vectors than from high-copy vectors and was enhanced by deletion of the downstream IS91 from plasmid inserts. Both core-linked (i.e., LPS) and non-core-linked (i.e., capsule-like) surface expression of form I O-Ps were detected by Western blotting and silver staining of polyacrylamide gel electrophoresis-separated Shigella and Escherichia coli extracts. However, salmonellae, which have a core that is chemically dissimilar to that of shigellae, expressed only non-core-linked surface-associated form I O-Ps. Finally, attenuated Salmonella enterica serovar Typhi live vaccine vector candidates, containing minimal-sized form I operon constructs, elicited immune protection in mice against virulent S. sonnei challenge, thereby supporting the promise of live, oral vaccines for the prevention of shigellosis. C1 US FDA, Ctr Food Safety & Nutr, Div Virulence Assessment, Laurel, MD 20708 USA. Univ Maryland, Sch Med, Dept Microbiol & Immunol, Baltimore, MD 21201 USA. US FDA, Ctr Biol Evaluat & Res, Lab Enter & Sexually Transmitted Dis, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Kopecko, DJ (reprint author), US FDA, Ctr Biol Evaluat & Res, Lab Enter & Sexually Transmitted Dis, HFM440,Bldg 29,Rm 420, Bethesda, MD 20892 USA. NR 46 TC 28 Z9 29 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 4414 EP 4423 DI 10.1128/IAI.70.8.4414-4423.2002 PG 10 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400050 PM 12117952 ER PT J AU Lei, BF Smoot, LM Menning, HM Voyich, JM Kala, SV Deleo, FR Reid, SD Musser, JM AF Lei, BF Smoot, LM Menning, HM Voyich, JM Kala, SV Deleo, FR Reid, SD Musser, JM TI Identification and characterization of a novel heme-associated cell surface protein made by Streptococcus pyogenes SO INFECTION AND IMMUNITY LA English DT Article ID GROUP-A STREPTOCOCCUS; CORYNEBACTERIUM-DIPHTHERIAE; BACTERICIDAL ANTIBODIES; NEISSERIA-MENINGITIDIS; BINDING; IRON; INFECTION; ACQUISITION; EXPRESSION; ANTIGEN AB Analysis of the genome sequence of a serotype M1 group A Streptococcus (GAS) strain identified a gene encoding a previously undescribed putative cell surface protein. The gene was cloned from a serotype M1 strain, and the recombinant protein was overexpressed in Escherichia coli and purified to homogeneity. The purified protein was associated with heme in a 1:1 stoichiometry. This streptococcal heme-associated protein, designated Shp, was produced in vitro by GAS, located on the bacterial cell surface, and accessible to specific antibody raised against the purified recombinant protein. Mice inoculated subcutaneously with GAS and humans with invasive infections and pharyngitis caused by GAS seroconverted to Shp, indicating that Shp was produced in vivo. The blood of mice actively immunized with Shp had significantly higher bactericidal activity than the blood of unimmunized mice. The shp gene was cotranscribed with eight contiguous genes, including homologues of an ABC transporter involved in iron uptake in gram-negative bacteria. Our results indicate that Shp is a novel cell surface heme-associated protein. C1 NIAID, Rocky Mt Lab, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Lab, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. OI DeLeo, Frank/0000-0003-3150-2516 NR 36 TC 72 Z9 72 U1 0 U2 7 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 4494 EP 4500 DI 10.1128/IAI.70.8.4494-4500.2002 PG 7 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400059 PM 12117961 ER PT J AU Kim, MK Seong, SY Seoh, JY Han, TH Song, HJ Lee, JE Shin, JH Lim, BU Kang, JS AF Kim, MK Seong, SY Seoh, JY Han, TH Song, HJ Lee, JE Shin, JH Lim, BU Kang, JS TI Orientia tsutsugamushi inhibits apoptosis of macrophages by retarding intracellular calcium release SO INFECTION AND IMMUNITY LA English DT Article ID RICKETTSIA-TSUTSUGAMUSHI; MURINE MACROPHAGES; CHEMOKINE GENES; INFECTION; LYMPHOCYTES; EXPRESSION; ACTIVATION; CELLS; MICE AB Orientia tsutsugamushi shows both pro- and antiapoptotic activities in infected vertebrate cells. Apoptosis of THP-1 cells induced by beauvericin was inhibited by O. tsutsugamushi infection. Beauvericin-induced calcium redistribution was significantly reduced and retarded in cells infected with O. tsutsugamushi. Antiapoptotic activities of O. tsutsugamushi in infected cells are most probably due to inhibition of the increase in the cytosolic calcium concentration. C1 Inha Univ, Coll Med, Dept Microbiol, Choong Gu, Inchon 400103, South Korea. Inha Univ, Coll Med, Dept Physiol, Inchon 400103, South Korea. Seoul Natl Univ, Coll Med, Dept Microbiol & Immunol, Seoul 110799, South Korea. Ewha Womans Univ, Coll Med, Dept Microbiol, Seoul 158710, South Korea. Sungkyunkwan Univ, Dept Microbiol & Immunol, Sch Med, Suwon 440746, South Korea. Jeollanamdo Inst Hlth & Environm, Div Microbiol, Kwangju 502810, South Korea. NIAID, Cellular & Mol Immunol Lab, NIH, Bethesda, MD 20894 USA. RP Kang, JS (reprint author), Inha Univ, Coll Med, Dept Microbiol, Choong Gu, Jungsuk B-D,3rd St, Inchon 400103, South Korea. EM jaeskang@inha.ac.kr RI Seong, Seung Yong /J-2764-2012; OI SEOH, JU-YOUNG/0000-0002-1168-5354 NR 24 TC 16 Z9 16 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 4692 EP 4696 DI 10.1128/IAI.70.8.4692-4696.2002 PG 5 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400083 PM 12117985 ER PT J AU Chen, W Palmer, RJ Kuramitsu, HK AF Chen, W Palmer, RJ Kuramitsu, HK TI Role of polyphosphate kinase in biofilm formation by Porphyromonas gingivalis SO INFECTION AND IMMUNITY LA English DT Article ID ESCHERICHIA-COLI; INORGANIC POLYPHOSPHATE; PSEUDOMONAS-AERUGINOSA; BACTERIAL PATHOGENS; STRAIN ADP1; GENE; VIRULENCE; MULTIPLE; PROTEIN; CLONING AB In order to assess the role of polyphosphate kinase (PPK) in the physiology of Porphyromonas gingivalis, a ppk gene mutant, CW120, was constructed and characterized. P. gingivalis was demonstrated to synthesize short-chain polyphosphate (polyP) but not long-chain polyp. CW120 failed to survive in the stationary phase as well as the parental cell did, and it was attenuated in biofilm formation on polyvinylchloride and glass surfaces. Furthermore, the complementation by insertion of an intact copy of the ppk gene into the mutant CW120 restored its biofilm formation and stationary-phase survival. These results suggest that PPK may be important for incorporation of these organisms into subgingival plaque in the human oral cavity. C1 SUNY Buffalo, Dept Oral Biol, Buffalo, NY 14214 USA. Natl Inst Dent & Craniofacial Res, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Kuramitsu, HK (reprint author), SUNY Buffalo, Dept Oral Biol, 3435 Main St, Buffalo, NY 14214 USA. FU NIDCR NIH HHS [DE08293] NR 45 TC 28 Z9 30 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD AUG PY 2002 VL 70 IS 8 BP 4708 EP 4715 DI 10.1128/IAI.70.8.4708-4715.2002 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 574VC UT WOS:000176909400087 PM 12117989 ER PT J AU Arbuckle, TE Burnett, R Cole, D Teschke, K Dosemeci, M Bancej, C Zhang, J AF Arbuckle, TE Burnett, R Cole, D Teschke, K Dosemeci, M Bancej, C Zhang, J TI Predictors of herbicide exposure in farm applicators SO INTERNATIONAL ARCHIVES OF OCCUPATIONAL AND ENVIRONMENTAL HEALTH LA English DT Article DE environmental exposure; herbicides; bio-monitoring; predictive modelling; exposure misclassification ID ASSESSING DERMAL EXPOSURE; VIDEO IMAGING TECHNIQUE; 2,4-DICHLOROPHENOXYACETIC ACID 2,4-D; PERCUTANEOUS PENETRATION; PESTICIDE EXPOSURE; OCCUPATIONAL EXPOSURE; EXCRETION; DIMETHYLAMINE; ABSORPTION; PREGNANCY AB Objectives: Most epidemiological studies of pesticides have used self-reports rather than quantitative measurements to assess exposures. The purpose of this study was to identify factors likely to affect exposure under actual field conditions and to measure the sensitivity and specificity of self-reported indications of exposure against urinary measures of herbicide exposure. Methods: A sub-set of the participants in a retrospective cohort study of Ontario farm families volunteered for a pesticide exposure assessment study. Immediately prior, and subsequent to, handling the phenoxy-herbicides 2,4-dichlorophenoxyacetic acid (2,4-D) or 4-chloro-2-methylphenoxyacetic acid (MCPA) for the first time during the season, 126 pesticide applicators provided pre-exposure spot urine samples and a subsequent consecutive 24-h urine sample. At the same time, they completed a questionnaire on pesticide use and handling practices for the first day of pesticide application. Results: Assuming that the presence of 2,4-D in the urine was a measure of true exposure and that questionnaire indications of 2,4-D use were the exposure classification subject to error, then the questionnaire's prediction of exposure had a sensitivity of 56.7% and specificity of 86.4%. The comparable values for MCPA were sensitivity and specificity of 91.6% and 67.4%, respectively. In multivariate models, the variables pesticide formulation, protective clothing/gear, application equipment, handling practice, and personal hygiene practice were significant as predictors of urinary herbicide levels in the first 24 h after application (or spraying) had been initiated (adjusted R2 44% for MCPA and 39% for 2,4-D). Conclusions: Although similar domains of factors were associated with exposure in both models, the specific factors identified and the signs of the coefficients were sufficiently different between the final models for each herbicide that additional investigations appear to be warranted to determine the sources of the differences and assess the validity of the models and their ability to be generalised. C1 Hlth Canada, Populat & Publ Hlth Branch, Hlth Surveillance & Epidemiol Div, Ottawa, ON K1A 0L2, Canada. Hlth Canada, Healthy Environm & Consumer Safety Branch, Off Policy & Programme Serv, Ottawa, ON K1A 0L2, Canada. Univ Toronto, Dept Publ Hlth Sci, Toronto, ON, Canada. Univ British Columbia, Dept Hlth Care & Epidemiol, Vancouver, BC V6T 1W5, Canada. US Natl Canc Inst, Occupat Epidemiol Branch, Rockville, MD USA. Hlth Canada, Canc Div, Ottawa, ON K1A 0L2, Canada. Hlth Canada, Bur Infect Dis, Ottawa, ON K1A 0L2, Canada. RP Arbuckle, TE (reprint author), Hlth Canada, Populat & Publ Hlth Branch, Hlth Surveillance & Epidemiol Div, AL 070ID, Ottawa, ON K1A 0L2, Canada. RI Cole, Donald /K-1040-2013 OI Cole, Donald /0000-0002-1009-603X NR 28 TC 65 Z9 66 U1 0 U2 5 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-0131 J9 INT ARCH OCC ENV HEA JI Int. Arch. Occup. Environ. Health PD AUG PY 2002 VL 75 IS 6 BP 406 EP 414 DI 10.1007/s00420-002-032307 PG 9 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 579JY UT WOS:000177175900006 PM 12070637 ER PT J AU Kagaya, A Takebayashi, M Okamura, H Saeki, T Morinobu, S Yamawaki, S Uchitomi, Y AF Kagaya, A Takebayashi, M Okamura, H Saeki, T Morinobu, S Yamawaki, S Uchitomi, Y TI Changes in mood and neurosteroids in women suffering from breast cancer SO INTERNATIONAL CLINICAL PSYCHOPHARMACOLOGY LA English DT Meeting Abstract CT Collegium Internationale Neuro-Psychopharmacologicum (CINP) CY OCT 02-05, 2001 CL HIROSHIMA, JAPAN C1 NIDA, Bethesda, MD 20892 USA. Hiroshima Univ, Sch Med, Dept Hlth Sci, Hiroshima 730, Japan. Hiroshima Univ, Sch Med, Dept Psychiat Neurosci, Hiroshima 730, Japan. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0268-1315 J9 INT CLIN PSYCHOPHARM JI Int. Clin. Psychopharmacol. PD AUG PY 2002 VL 17 SU 2 BP S31 EP S32 PG 2 WC Pharmacology & Pharmacy; Psychiatry SC Pharmacology & Pharmacy; Psychiatry GA 614QA UT WOS:000179199800068 ER PT J AU van Gils, CH Conway, K Li, Y Taylor, JA AF van Gils, CH Conway, K Li, Y Taylor, JA TI HRAS1 variable number of tandem repeats polymorphism and risk of bladder cancer SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE bladder cancer; polymorphism; HRAS1; variable number tandem repeat (VNTR) ID RARE MINISATELLITE ALLELES; AGE 40 YEARS; BREAST-CANCER; OVARIAN-CANCER; GENETIC SUSCEPTIBILITY; AUSTRALIAN WOMEN; LOCUS; SEQUENCE; TUMORS AB The HRAS1 variable number of tandem repeats (VNTR) polymorphism, 1 kb downstream from the HRAS1 gene, has been reported to be associated with risk of various cancers. To examine whether individuals with rare HRAS1 VNTR alleles are at increased risk of bladder cancer we carried out a case control study with 230 bladder cancer cases and 203 hospital-based controls frequency-matched on ethnicity, gender and age. For genotyping we used a PCR-based long-gel electrophoretic assay that provides precise allele size discrimination. We did not find evidence of a strong overall effect of the HRAS1 VNTR on bladder cancer risk. Genotype data for whites and blacks were analyzed separately, but the number of black subjects was too small to estimate meaningful odds ratios. Compared to white subjects with 2 common alleles, the odds ratio (OR) for white subjects with I rare allele was 0.9 (95% confidence interval (Cl) 0.5-1.4) and for those with 2 rare alleles OR = 1.7 (95% Cl 0.6-5.4). HRASI genotype may be related to the prognosis of bladder cancer, however, because incident cases, i.e., newly diagnosed cases had a higher frequency of rare alleles than did prevalent cases, i.e., cases already existing at the time of recruitment. Repeating the analyses with incident cases only (n = 53), the OR for subjects with I rare allele was 1.2 (95% Cl = 0.6-2.4) and for those with 2 rare alleles 3.2 (95% CI = 0.8-13.7). The number of incident cases was too small to draw firm conclusions on a possible association with a subgroup of tumors with a poor prognosis. Published 2002 Wiley-Liss, Inc.(dagger) C1 NIEHS, Mol & Genet Epidemiol Sect, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, Chapel Hill, NC USA. RP Taylor, JA (reprint author), NIEHS, Mol & Genet Epidemiol Sect, Mol Carcinogenesis Lab, NIH, POB 12233,MD A3-05, Res Triangle Pk, NC 27709 USA. OI taylor, jack/0000-0001-5303-6398 NR 26 TC 6 Z9 6 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD AUG 1 PY 2002 VL 100 IS 4 BP 414 EP 418 DI 10.1002/ijc.10497 PG 5 WC Oncology SC Oncology GA 570JE UT WOS:000176654500006 PM 12115522 ER PT J AU Clement, PMJ Hanauske-Abel, HM Wolff, EC Kleinman, HK Park, MH AF Clement, PMJ Hanauske-Abel, HM Wolff, EC Kleinman, HK Park, MH TI The antifungal drug ciclopirox inhibits deoxyhypusine and proline hydroxylation, endothelial cell growth and angiogenesis in vitro SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE ciclopirox; angiogenesis; hypusine; HUVEC; eIF5A; deoxyhypusine hydroxylase; prolyl 4-hydroxylase; deferiprone ID INITIATION-FACTOR 5A; YEAST SACCHAROMYCES-CEREVISIAE; HYPUSINE FORMATION; DNA-REPLICATION; PROTEIN-SYNTHESIS; MAMMALIAN-CELLS; IRON CHELATORS; RAT TESTIS; EIF-5A; MIMOSINE AB The hypusine biosynthetic steps represent novel targets for intervention in cell proliferation. Hypusine is a rare amino acid, formed posttranslationally in one cellular protein, eIF5A, and is essential for cell proliferation. Deoxyhypusine hydroxylase, the metalloenzyme catalyzing the final step in hypusine biosynthesis, and prolyl 4-hydroxylase, a non-heme iron enzyme critical for collagen processing, can be inhibited by small chelating molecules that target their essential metal atom. We examined the effects of 5 compounds (ciclopirox, deferiprone, deferoxamine, mimosine and 2,2'-dipyridyl) on these protein hydroxylases in HUVECs, on cell proliferation and on angiogenesis using 2 model assays: tube-like vessel formation on Matrigel and the chick aortic arch sprouting assay. These compounds inhibited cellular deoxyhypusine hydroxylase in a concentration-dependent manner, but their efficacy varied widely in the following order: ciclopirox-->deferoxamine-2,2'-dipyridyl -->deferiprone-->mimosine (IC50 5-200 muM). Inhibition of DNA synthesis, following the same order (IC50 10-450 muM), correlated with G, arrest of the cell cycle. These compounds also inhibited proline hydroxylation and maturation of collagen in HUVECs and caused inhibition of angiogenesis in vitro. Of the compounds tested, ciclopirox was by far the most effective inhibitor of HUVEC proliferation and angiogenesis. The strong antiangiogenic activity of this readily available antifungal drug along with its antiproliferative effects suggests a new potential application for ciclopirox in the treatment of solid tumors. Published 2002 Wiley-Liss, Inc.(dagger). C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD USA. RP Park, MH (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bldg 30,Rm 211, Bethesda, MD 20892 USA. NR 66 TC 73 Z9 76 U1 0 U2 5 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD AUG 1 PY 2002 VL 100 IS 4 BP 491 EP 498 DI 10.1002/ijc.10515 PG 8 WC Oncology SC Oncology GA 570JE UT WOS:000176654500020 PM 12115536 ER PT J AU Orlic, D AF Orlic, D TI Stem cell repair in ischemic heart disease: An experimental model SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article; Proceedings Paper CT 29th Wolrd Congress of the International-Society-of-Hematology CY 2002 CL SEOUL, SOUTH KOREA SP Int Soc Hematol ID MYOCARDIUM AB Bone marrow stem cells (BMSC) from adult mice are now believed to generate non-hematopoietic cell types. This newly defined property is referred to as stem cell plasticity. We tested the potential of lineage negative c-kit positive (Lin- c-kit(+)), GFP+ BMSC to differentiate into cardiac myocytes in myocardial infarcts produced by ligation of the left coronary artery. At 9 days post-transplant the hearts showed a band of developing GFP+ myocytes within the damaged myocardium. These GFP+ myocytes were positive for cardiac specific myosin and early expressed transcription factors. Endothelial cells and smooth muscle cells also developed from the donor bone marrow cells. Left ventricular end diastolic pressure (LVEDP) and left ventricular developed pressure (LVDP) were improved. Lin-c-kit- cells did not regenerate myocardium. We next tested the ability of cytokine-mobilized BMSC to regenerate myocardium. Nuclei in regenerating cardiomyocytes were positive for Csx/Nkx 2.5, GATA-4 and MEF2. Cytoplasmic proteins included desmin, nestin and connexin 43. Regenerating arterioles consisted of endothelial cells and smooth muscle cells positive for Ki67, and flk1. These regenerating vessels contained circulating TER119 positive red blood cells. Repair of infarcted myocardium resulted in improved heart function and survival. At day 27 after cytokine treatment and surgery, 11 of 15 mice survived compared with 9 of 52 non-treated mice. Left ventricular ejection fraction in infarcted hearts in cytokine-treated mice was 48%, 62% and 114% higher than the ejection fraction in non-treated mice at 9, 16 and 26 days following coronary artery occlusion. These findings demonstrate that circulating autologous stem cells traffic to the ischemic, infarcted myocardium and undergo differentiation into cardiomyocytes and vascular structures. We conclude that adult BMSC have the potential for repair in acute, ischemic heart disease. C1 NHGRI, NIH, Bethesda, MD 20892 USA. RP Orlic, D (reprint author), NHGRI, NIH, Bethesda, MD 20892 USA. NR 5 TC 23 Z9 31 U1 1 U2 7 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD AUG PY 2002 VL 76 SU 1 BP 144 EP 145 PG 2 WC Hematology SC Hematology GA 611FM UT WOS:000179005800004 PM 12430844 ER PT J AU Schmidt, M Hacein-Bey, S Carbonaro, D Wissler, M LeDeist, F Cavazzano-Calvo, M Kohn, DB Dunbar, C Fischer, A von Kalle, C AF Schmidt, M Hacein-Bey, S Carbonaro, D Wissler, M LeDeist, F Cavazzano-Calvo, M Kohn, DB Dunbar, C Fischer, A von Kalle, C TI Insight into hematopoiesis from primate and human gene transfer studies SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article; Proceedings Paper CT 29th Wolrd Congress of the International-Society-of-Hematology CY 2002 CL SEOUL, SOUTH KOREA SP Int Soc Hematol C1 Univ Freiburg, Freiburg, Germany. Hop Necker Enfants Malad, Unite Immunol & Hematol Pediat, Paris, France. Childrens Hosp Los Angeles, Div Immunol Res, BMT, Los Angeles, CA USA. Univ So Calif, Keck Sch Med, Dept Pediat, Los Angeles, CA 90089 USA. Univ So Calif, Keck Sch Med, Dept Microbiol, Los Angeles, CA 90089 USA. NHLBI, NIH, Bethesda, MD 20892 USA. RP Schmidt, M (reprint author), Univ Freiburg, Freiburg, Germany. NR 0 TC 0 Z9 0 U1 0 U2 0 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD AUG PY 2002 VL 76 SU 1 BP 148 EP 148 PG 1 WC Hematology SC Hematology GA 611FM UT WOS:000179005800006 ER PT J AU Young, NS Maciejewski, JP Sloand, E Chen, GB Zeng, WH Risitano, A Miyazato, A AF Young, NS Maciejewski, JP Sloand, E Chen, GB Zeng, WH Risitano, A Miyazato, A TI The relationship of aplastic anemia and PNH SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article ID PAROXYSMAL-NOCTURNAL HEMOGLOBINURIA; PIG-A GENE; T-CELL REPERTOIRE; BONE-MARROW; ANCHORED PROTEINS; SOMATIC MUTATIONS; MUTANT-CELLS; DEFICIENT; HEMATOPOIESIS; CYCLOSPORINE AB Bone marrow failure has been regarded as one of the triad of clinical manifestations of paroxysmal nocturnal hemoglobinuria (PNH), and PNH in turn has been described as a late clonal disease evolving in patients recovering from aplastic anemia. Better understanding of the pathophysiology of both diseases and improved tests for cell surface glycosylphosphatidylinositol (GPI)-linked proteins has radically altered this view. Flow cytometry of granulocytes shows evidence of an expanded PNH clone in a large proportion of marrow failure patients at the time of presentation: in our large NIH series, about 1/3 of over 200 aplastic anemia cases and almost 20% of more than 100 myelodysplasia cases. Clonal PNH expansion (rather than bone marrow failure) is strongly linked to the histocompatability antigen HLA.-DR2 in all clinical varieties of the disease, suggesting an immune component to its pathophysiology. An extrinsic mechanism of clonal expansion is also more consistent with knock-out mouse models and culture experiments with primary cells and cell lines, which have failed to demonstrate an intrinsic proliferative advantage for PNH cells. DNA chip analysis of multiple paired normal and PIG-A mutant cell lines and lymphoblastoid cells do not show any consistent differences in levels of gene expression. In aplastic anemia/PNH there is surprisingly limited utilization of the V-beta chain of the T cell receptor, and patients' dominant T cell clones, which are functionally inhibitory of autologous hematopoiesis, use identical CDR3 regions for antigen binding. Phenotypically normal cells from PNH patients proliferate more poorly in culture than do the same patient's PNH cells, and the normal cells are damaged as a result of apoptosis and overexpress Fas. Differences in protein degradation might play a dual role in pathophysiology, as GPI-linked proteins lacking an anchor would be predicted to be processed by the proteasome machinery and displayed in a class I H.A. context, in contrast to the normal pathway of cell surface membrane recycling, lysosomal degradation, and presentation by class II HLA. The strong relationship between a chronic, organ-specific immune destructive process and the expansion of a single mutant stem cell clone remains frustratingly enigmatic but likely to be the result of interesting biologic processes, with mechanisms that potentially can be extended to the role of inflammation in producing premalignant syndromes. C1 NHLBI, Hematol Branch, Bethesda, MD 20892 USA. RP Young, NS (reprint author), NHLBI, Hematol Branch, Bethesda, MD 20892 USA. NR 40 TC 21 Z9 21 U1 0 U2 2 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD AUG PY 2002 VL 76 SU 2 BP 168 EP 172 PG 5 WC Hematology SC Hematology GA 611FP UT WOS:000179006000036 PM 12430920 ER PT J AU Maciejewski, JP Risitano, A Kook, H Zeng, WH Chen, GB Young, NS AF Maciejewski, JP Risitano, A Kook, H Zeng, WH Chen, GB Young, NS TI Immune pathophysiology of aplastic anemia SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article; Proceedings Paper CT 29th Wolrd Congress of the International-Society-of-Hematology CY 2002 CL SEOUL, SOUTH KOREA SP Int Soc Hematol ID T-CELL RECEPTOR; VERSUS-HOST DISEASE; PAROXYSMAL-NOCTURNAL HEMOGLOBINURIA; CHAIN VARIABLE REGIONS; BONE-MARROW FAILURE; V-BETA REPERTOIRE; MYELODYSPLASTIC SYNDROME; RHEUMATOID-ARTHRITIS; HEMATOPOIETIC-CELLS; CLONAL EXPANSIONS AB Aplastic anemia (AA) remains an elusive disease. Its pathophysiology is not only fascinating by the seemingly simple findings of cytopenia and marrow hypoplasia, but may also contain key information to the understanding of other fundamental processes such as stem cell regeneration, evolution, and immune control of clonal diseases. Although measurements of blood counts provide an objective tool to assess the disease activity and response to the therapy, immune pathophysiology of AA, as inferred from the successes of immunosuppression, provides only few other clinical clues. Similarly, the current laboratory evidence remains mostly indirect. In spite of the recognition of immune pathways of hematopoietic inhibition and apoptosis in AA, the fundamental question about the nature of the antigen(s) inciting or maintaining the pathologic immune response that ultimately leads to bone marrow failure, remains open. However, recognition of the immune targets may aid in understanding not only the pathogenesis but also many of clinical associations and the late squelae of AA. For example, abnormal cells in AA and myelodysplastic syndrome (MDS) MDS may harbor inciting antigens but the immune response lacks selectivity. Clonal selection pressure may be a result of this process or alternatively, emergence of tolerance could lead to the establishment of abnormal hematopoiesis. Clonal proliferation of large granular lymphocytosis could represent an example of an exaggerated response to an immunodominant hematopoietic antigen. In addition to the traditional functional or phenotypic analysis, pathologic immune response in AA can be studied on molecular level by identifying and quantitating T cell clones based on the presence of unique variable B-chain CDR3 sequences. Detection of clonal expansion is based on the observation that in infections and autoimmune conditions, the presence of antigenic drive will lead to the expansion and overrepresentation of T cell clones recognizing this antigen. However, simple analysis of clonal representation is not sufficient to resolve the complex nature of the immune repertoire in the context of genetic and clinical heterogeneity. Therefore, we analyzed VB and CDR3 repertoire in CD4 and CD8 cells, activated or effector cell subsets. To distinguish truly expanded and likely immunodominant clones, we first studied VB distribution and cloned CDR3 sequences from expanded VB families. Identified clonotypic sequences can be used to design molecular tests to quantitate the strength of pathologic immune response. Clonotype sharing has been confirmed in patients with similar clinical features indicating presence of common antigens. In addition, quantitative analysis showed correlation with the therapy response. Persistence and patterns of clonotypes may be helpful in the classification of immune-mediated marrow failure based on the immune characteristics and will allow inferences into the inciting pathways. C1 Cleveland Clin Fdn, Taussig Canc Ctr, Expt Hematol & Hematopoiesis Sect, Cleveland, OH 44195 USA. NHLBI, Hematol Branch, Bethesda, MD 20892 USA. RP Maciejewski, JP (reprint author), Cleveland Clin Fdn, Taussig Canc Ctr, Expt Hematol & Hematopoiesis Sect, 9500 Euclid Ave, Cleveland, OH 44195 USA. NR 77 TC 16 Z9 23 U1 0 U2 2 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD AUG PY 2002 VL 76 SU 1 BP 207 EP 214 PG 8 WC Hematology SC Hematology GA 611FM UT WOS:000179005800024 PM 12430855 ER PT J AU Kwak, LW AF Kwak, LW TI Translational development of active immunotherapy for hematologic malignancies SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article; Proceedings Paper CT 29th Wolrd Congress of the International-Society-of-Hematology CY 2002 CL SEOUL, SOUTH KOREA SP Int Soc Hematol ID VACCINATION; LYMPHOCYTES; IMMUNITY; PROTEIN; CANCER; CELLS AB The B-cell tumor-derived Ig receptor may be considered a model tumor antigen for cancer vaccine development. However, as a non-immunogenic, self-antigen, it also must be first rendered immunogenic by chemical or genetic fusion to carriers which enable the induction of protective antitumor immunity in experimental tumor models. Our group has demonstrated that active immunization of human patients with idiotypic protein vaccines containing soluble GM-CSF elicited antigen specific CD8(+) T cell responses and antitumor effects. An alternative strategy to develop vaccines is the genetic fusion of tumor idiotype-derived single chain antigen with a chemokine moiety. Administration of these vaccines as fusion proteins or naked DNA vaccines may allow more efficient targeting of antigen presenting cells in vivo. Potent antitumor immunity was elicited in mice which was dependent on the generation of specific antibodies and both CD4(+) and CD8(+) effector T-cells. We propose that chemokine fusion may represent a novel, general strategy for formulating existing or newly identified tumor and HIV antigens into vaccines for cancer and AIDS, respectively, which elicit potent CD8(+) T-cell immunity. C1 NIH, Bethesda, MD 20892 USA. RP Kwak, LW (reprint author), NIH, Bethesda, MD 20892 USA. NR 8 TC 0 Z9 0 U1 0 U2 0 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD AUG PY 2002 VL 76 SU 1 BP 320 EP 321 PG 2 WC Hematology SC Hematology GA 611FM UT WOS:000179005800053 PM 12430873 ER PT J AU Field, JK Brambilla, C Caporaso, N Flahault, A Henschke, C Herman, J Hirsch, F Lachmann, P Lam, S Maier, S Montuenga, LM Mulshine, J Murphy, M Pullen, J Spitz, M Tockman, M Tyndale, R Wistuba, I Youngson, J AF Field, JK Brambilla, C Caporaso, N Flahault, A Henschke, C Herman, J Hirsch, F Lachmann, P Lam, S Maier, S Montuenga, LM Mulshine, J Murphy, M Pullen, J Spitz, M Tockman, M Tyndale, R Wistuba, I Youngson, J TI Consensus statements from the Second International Lung Cancer Molecular Biomarkers Workshop: A European strategy for developing lung cancer molecular diagnostics in high risk populations SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE lung cancer; consensus statement; workshop report; Spiral CT; molecular biomarkers; sputum; validation; ethics AB The Second Molecular Biomarkers Workshop was held at the Roy Castle International Centre for Lung Cancer Research in Liverpool, in June 2001 and it brought together experts in the clinical, epidemiological and molecular-pathology of lung cancer from Europe and the USA, to address issues surrounding the development of a European strategy for early lung cancer detection. The 2001 Workshop Breakout Groups concentrated on the current challenges in the early detection of lung cancer which need to be addressed in the light of the recent surge in interest in many countries for mounting new clinical trials to evaluate the utility of Spiral CT in early lung cancer detection. If population-based trials of CT screening are mounted it will also be a favorable clinical environment in which to evaluate efficiently recent advances in molecular screening and genotyping. The Workshop focused specifically on: a) clinical and molecular biomarkers, b) sputum as an early detection and diagnostic tool, c) validation of molecular markers prior to their use in early detection trials and d) ethical issues that have to be considered in early lung cancer detection trials. A distillation of the Workshop discussions is given in this article. C1 Univ Liverpool, Roy Castle Int Ctr Lung Canc Res, Liverpool L3 9TA, Merseyside, England. Univ Grenoble, Ctr Hosp, Serv Pneumol, Grenoble, France. NCI, Genet Epidemiol Branch, Div Canc Epidemiol & Genet, Rockville, MD 20852 USA. INSERM, U444, WHO, Collaborating Ctr Elect Dis Surveillance, F-75571 Paris, France. New York Presbyterian Hosp, Dept Radiol, New York, NY 10021 USA. Johns Hopkins Oncol Ctr, Sch Med, Baltimore, MD 21287 USA. Univ Colorado, Hlth Sci Ctr, Dept Pathol, Denver, CO 80262 USA. Univ Cambridge, Microbial Immunol Grp, Ctr Vet Sci, Cambridge CB3 0ES, England. British Columbia Canc Agcy, Vancouver, BC V5Z 4E6, Canada. Epigenom AG, D-10435 Berlin, Germany. Univ Navarra, Sch Med, Dept Histol & Pathol, E-31080 Pamplona, Spain. NCI, Bethesda, MD 20892 USA. Inst Hlth Sci, Imperial Canc Res Fund, Gen Practice Res Grp, Dept Primary Hlth Care, Oxford OX3 7LF, England. Oxagen Ltd, Abingdon OX14 4RY, Oxon, England. Univ Texas, MD Anderson Canc Ctr, Texas Med Ctr, Dept Epidemiol, Houston, TX 77030 USA. Univ S Florida, H Lee Moffitt Canc Ctr, Mol Screening Program, Tampa, FL 33612 USA. Univ Toronto, Toronto, ON M5S 1A8, Canada. Catholic Univ Chile, Dept Pathol, Santiago, Chile. RP Field, JK (reprint author), Univ Liverpool, Roy Castle Int Ctr Lung Canc Res, 200 London Rd, Liverpool L3 9TA, Merseyside, England. OI Field, John/0000-0003-3951-6365 NR 2 TC 15 Z9 15 U1 0 U2 1 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD AUG PY 2002 VL 21 IS 2 BP 369 EP 373 PG 5 WC Oncology SC Oncology GA 574WQ UT WOS:000176913000019 PM 12118333 ER PT J AU Ciolino, HP Dankwah, M Yeh, GC AF Ciolino, HP Dankwah, M Yeh, GC TI Resistance of MCF-7 cells to dimethylbenz(a)anthracene-induced apoptosis is due to reduced CYP1A1 expression SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE CYP1A1; aryl hydrocarbon receptor; DMBA; apoptosis; MCF-7 ID ARYL-HYDROCARBON RECEPTOR; CYTOCHROME-P450 1A1; GENE; BENZOPYRENE; TRANSCRIPTION; INDUCTION; MOUSE; MECHANISMS; ACTIVATION; CASPASE-3 AB We have developed a series of aryl hydrocarbon (AH)-resistant cell lines derived from MCF-7 human breast epithelial cancer cells by continuous exposure to the AH benzo[a]pyrene. These cell lines display cross-resistance to the mammary carcinogen dimethylbenz[a]anthracene (DMBA). Apoptosis induced by exposure to DMBA is greatly decreased in the resistant cell lines compared to the wild-type, in proportion to the level of resistance. Apoptosis induced by DMBA could be blocked by inhibitors of DMBA metabolism such as a-naphthoflavone and diosmetin. We therefore examined the resistant cell lines for their ability to metabolize DMBA and for the formation of DMBA-DNA adducts, and found that both parameters were decreased compared to wild-type cells in proportion to the level of resistance. When exposed to DMBA or 2,3,7,8-tetrachlorodibenzo-p-dioxin, the resistant cell lines have a diminished capacity to carry out ethoxyresorufin-O-deethylation, indicating that the induction of cytochrome P450 1A1 (CYP1A1) enzyme is impaired. We therefore examined the expression of the CYP1A1 gene, and found reduced levels of both CYP1A1 mRNA and CYP1A1-promoter controlled transcription in resistant cells compared to the wild-type. The deleterious effects of AHs are believed to be mediated by the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor which regulates CYP1A1 expression. Resistant cell lines had a reduced expression of the AhR, as measured at the mRNA and protein levels. These data demonstrate that AH resistance in these cells is mediated by changes in the signal transduction pathway which regulates CYP1A1 expression. C1 NCI, Cellular Def & Carcinogenesis Sect, Basic Res Lab, Ctr Canc Res,NIH, Frederick, MD 21702 USA. RP Yeh, GC (reprint author), NCI, Cellular Def & Carcinogenesis Sect, Basic Res Lab, Ctr Canc Res,NIH, Bldg 560 Room 12-05, Frederick, MD 21702 USA. NR 39 TC 21 Z9 25 U1 0 U2 2 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD AUG PY 2002 VL 21 IS 2 BP 385 EP 391 PG 7 WC Oncology SC Oncology GA 574WQ UT WOS:000176913000022 PM 12118336 ER PT J AU Gunji, T Kawauchi, N Akahane, M Watanabe, K Kanamori, H Ohnishi, S AF Gunji, T Kawauchi, N Akahane, M Watanabe, K Kanamori, H Ohnishi, S TI Long-term outcomes of transcatheter arterial chemoembolization with autologous blood clot for unresectable hepatocellular carcinoma SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE hepatocellular carcinoma; transcatheter arterial chemoembolization; autologous blood clot ID PERCUTANEOUS ETHANOL INJECTION; PROGNOSTIC FACTORS; RANDOMIZED TRIAL; CIRRHOSIS; DOXORUBICIN; LIVER; ULTRASONOGRAPHY; EMBOLIZATION; MANAGEMENT; CISPLATIN AB Ninety-two consecutive patients with unresectable hepatocellular carcinoma (HCC) greater than 2 cm and less than 5 cm in diameter were treated by transcatheter arterial chemoembolization using autologous blood clot as an embolizing agent (Short-TAE; S-TAE). Survival, patency of hepatic arteries and side effects were retrospectively analyzed. The median follow-up interval was 42 months. The median size of the main tumor was 3.4 cm in diameter. All tumors were determined to be inoperable because of intrahepatic spread of tumors and/or poor hepatic functional reserves. S-TAE was performed by injecting a mixture of iodized oil and anticancer drugs followed by embolization of hepatic arteries with autologous blood clot. Embolization with clots maintained patency of hepatic arteries in all the patients. The overall survival rates for 92 HCC patients at 1, 3, 5, 7 and 8 years were estimated by the Kaplan-Meier method to be 100, 52, 34, 12 and 6%, respectively, which were better compared with prior records for conventional method with gelfoam. S-TAE was successfully performed in 16 patients with Childs class C liver cirrhosis presenting icterus and ascitis. The survival rates for Childs class C patients at 1, 2 and 3 years were 100, 66 and 32%, respectively. The Cox proportional hazard model was used to assess influence of each pretreatment parameter (age, gender, virus marker, stage of cirrhosis, number of tumor) on survival. The univariate analysis demonstrated that both the stage of cirrhosis and the multiplicity of tumor were significant factors predicting survival (p=0.003 and 0.046, respectively). The multivariate analysis showed the stage of cirrhosis to be the sole factor which significantly affected prognosis of HCC patients (p=0.005). In conclusion, chemoembolization with autologous blood clot is an effective and safe therapeutic option for unresectable HCC smaller than 5 cm. The hepatic functional reserves play a key role in determining prognosis of HCC patients. C1 Univ Tokyo, Fac Med, Dept Internal Med 3, Tokyo 113, Japan. Univ Tokyo, Fac Med, Dept Radiol, Tokyo 113, Japan. RP Gunji, T (reprint author), NIDDK, Liver Dis Sect, Digest Dis Branch, NIH, Bldg 10,Rm 9B06,10 Ctr Dr,MSC 1800, Bethesda, MD 20892 USA. NR 25 TC 10 Z9 10 U1 0 U2 0 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD AUG PY 2002 VL 21 IS 2 BP 427 EP 432 PG 6 WC Oncology SC Oncology GA 574WQ UT WOS:000176913000027 PM 12118341 ER PT J AU Stern, ME Gao, JP Schwalb, TA Ngo, ML Tieu, DD Chan, CC Reis, BL Whitcup, SM Thompson, D Smith, JA AF Stern, ME Gao, JP Schwalb, TA Ngo, ML Tieu, DD Chan, CC Reis, BL Whitcup, SM Thompson, D Smith, JA TI Conjunctival T-cell subpopulations in Sjogren's and non-Sjogren's patients with dry eye SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID KERATOCONJUNCTIVITIS SICCA; LACRIMAL GLANDS; SALIVARY-GLAND; IMMUNE-SYSTEM; ASSOCIATION; EXPRESSION; EPITHELIUM; RECEPTORS; THERAPY AB PURPOSE. To examine the conjunctiva of patients with Sjogren's syndrome keratoconjunctivitis sicca (SS-KCS) and non-Sjogren's keratoconjunctivitis sicca (NS-KCS) for evidence of immune-based inflammation. METHODS. Conjunctival biopsy specimens were obtained from 15 patients with SS-KCS and 15 with NS-KCS. Immunohistochemistry was performed on frozen sections to characterize and quantify T-cell subtypes (CD3, CD4, and CD8) and markers, of immune activation (major histocompatibility complex [MHC] class II: HLA-DR, HLA-DQ) and inflammation (intercellular adhesion molecule [ICAM]-1), The numbers of cells positive for each market were counted by two masked observers and averaged. RESULTS. Conjunctival biopsy specimens from patients with SS-KCS or NS-KCS revealed lymphocytic infiltration and increased immunoreactivity for the markers of inflammation and immune activation. The extent of cellular immunoreactivity did not differ significantly between SS-KCS and NS-KCS tissue samples. CONCLUSIONS. The authors' findings indicate that patients with SS-KCS or NS-KCS have conjunctival inflammation manifested by inflammatory cell infiltrates and upregulation of expression in markers of immune activation. Clinical symptoms of KCS may be more dependent on T-cell activation and resultant inflammation than previously believed. In addition to tear substitutes, anti-inflammatory therapeutics should be investigated for the treatment of KCS. C1 Allergan Pharmaceut Inc, Irvine, CA 92612 USA. NEI, Clin Branch, Bethesda, MD USA. NEI, Lab Immunol, Bethesda, MD USA. EMMES Corp, Potomac, MD USA. RP Stern, ME (reprint author), Allergan Pharmaceut Inc, 2525 Dupont Dr,RD-2C, Irvine, CA 92612 USA. NR 21 TC 170 Z9 175 U1 1 U2 4 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD AUG PY 2002 VL 43 IS 8 BP 2609 EP 2614 PG 6 WC Ophthalmology SC Ophthalmology GA 577ZZ UT WOS:000177094100015 PM 12147592 ER PT J AU Takase, H Sugita, S Rhee, D Imai, Y Taguchi, C Sugamoto, Y Tagawa, Y Nishihira, J Russell, P Mochizuki, M AF Takase, H Sugita, S Rhee, D Imai, Y Taguchi, C Sugamoto, Y Tagawa, Y Nishihira, J Russell, P Mochizuki, M TI The presence of macrophage migration inhibitory factor in human trabecular meshwork and its upregulatory effects on the T helper 1 cytokine SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID SOLUBLE FAS LIGAND; AQUEOUS-HUMOR; IMMUNOHISTOCHEMICAL LOCALIZATION; MATRIX METALLOPROTEINASES; GLAUCOMATOUS EYES; UVEITIS PATIENTS; ORGAN-CULTURE; OCULAR FLUIDS; EXPRESSION; OUTFLOW AB PURPOSE. To investigate the expression and secretion of macrophage migration inhibitors factor (MIF) in human trabecular meshwork (HTM) and evaluate its role in ocular inflammation. METHODS. Tissue samples of HTM cells were isolated from donor human eyes or corneoscleral buttons, and the HTM cells were cultured. The expression of MIF on HTM cells was evaluated by RT-PCR, Western blot analysis, and ELISA. T-cell clones (TCCs) were established from Ocular infiltrating cells of patients with uveitis. ELISA was used to evaluate the pathologic role of MIF, in relation to regulatory effects on cytokine production by T cells. RESULTS. MIF was detected in the HTM by RT-PCR and Western blot analysis. MIF was also shown by ELISA to be secreted by the HTM cells in culture. The HTM supernatant enhanced IFN-gamma production by TCCs, but not IL-10; and these effects were neutralized by anti-MIF antibodies, Similarly, recombinant MIF enhanced the IFN-gamma production by the TCCs. CONCLUSIONS. MIF is expressed and secreted in the HTM, and MIF has the capacity to enhance T helper 1 cytokines and may play a role as an inflammatory cytokine in the eye. C1 Tokyo Med & Dent Univ, Grad Sch, Dept Ophthalmol & Visual Sci, Bunkyo Ku, Tokyo 1138519, Japan. NEI, Lab Mechanisms Ocular Dis, NIH, Bethesda, MD 20892 USA. Kurume Univ, Sch Med, Dept Ophthalmol, Kurume, Fukuoka 830, Japan. Hokkaido Univ, Sch Med, Dept Ophthalmol, Sapporo, Hokkaido 060, Japan. Hokkaido Univ, Sch Med, Cent Res Inst, Sapporo, Hokkaido 060, Japan. RP Mochizuki, M (reprint author), Tokyo Med & Dent Univ, Grad Sch, Dept Ophthalmol & Visual Sci, Bunkyo Ku, 1-5-45 Yushima, Tokyo 1138519, Japan. NR 39 TC 11 Z9 13 U1 0 U2 0 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD AUG PY 2002 VL 43 IS 8 BP 2691 EP 2696 PG 6 WC Ophthalmology SC Ophthalmology GA 577ZZ UT WOS:000177094100027 PM 12147604 ER PT J AU Sakurai, S Alam, S Pagan-Mercado, G Hickman, F Tsai, JY Zelenka, P Sato, S AF Sakurai, S Alam, S Pagan-Mercado, G Hickman, F Tsai, JY Zelenka, P Sato, S TI Retinal capillary pericyte proliferation and c-fos mRNA induction by prostaglandin D-2 through the cAMP response element SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID ENDOTHELIAL GROWTH-FACTOR; SMOOTH-MUSCLE CELLS; SYNTHASE BETA-TRACE; ANGIOTENSIN-II; TRANSCRIPTIONAL ACTIVATION; DIABETIC-RETINOPATHY; FACTOR EXPRESSION; IN-VIVO; ANGIOGENESIS; CYCLOOXYGENASE-2 AB PURPOSE. Cycloxygenase inhibitors have been shown to pre vent angiogenesis in some circumstances, suggesting that growth of capillary pericytes or endothelial cells may be regulated by prostaglandins (PGs). The present study tests the effects of PGs on the growth of human retinal capillary pericytes. METHODS. Cell growth was assayed by formazan formation and 5-bromo-2'-deoxyuridine (BrdU) incorporation. The expression of mRNAs corresponding to c-fos, PG receptors, and VEGF was examined by RT-PCR. Signal transduction was evaluated by immunoblot analysis using phosphospecific antibodies against mitogen-activated protein kinases (MAPKs) and cAMP response element-binding protein (CREB). Svnthesis of CAMP was inhibited with the adenyl cyclase inhibitor SQ22536. A reporter gene (luciferase) assay was conducted using the expression vector pSVOADelta5' containing the 379-bp c:fos promoter with and without a mutation in CAMP response element (CRE). RESULTS. PGD(2) treatment induced c-fos mRNA, stimulated pericyte growth, and increased expression of VEGF mRNA. PGE(2) and -F-2alpha. had similar effects on c-fos induction and pericyte growth, whereas PGI(2) was ineffective, RT-PCR confirmed that mRNAs corresponding to the receptors for PGD(2),-E2, -F-2alpha and 42 were expressed in human retinal pericytes. Stimulation bv, PGD2 led to phosphorylation of CREB, but had negligible effect on phosphorylation of p44/42 MAPK. The adenylyl cyclase inhibitor inhibited CREB activation and c:fos in diction by PGD2. In a reporter gene assay, c-fos induction occurred only with wild-type c-fos promoter. Mutation in CRE eliminated the response to PGD(2). CONCLUSIONS. PGD2 promotes the growth of retinal capillary pericytes by signaling through CAMP and CREB. The findings underscore the importance of PGs in the growth of human retinal capillary pericytes and raise the possibility that PGs may play a role in proliferative retinopathies. C1 Univ Oklahoma, Hlth Sci Ctr, Dept Med, Oklahoma City, OK 73190 USA. NEI, Lab Ocular Therapeut, NIH, Bethesda, MD 20892 USA. NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Sato, S (reprint author), Univ Oklahoma, Hlth Sci Ctr, Dept Med, POB 26901,BSEB 331, Oklahoma City, OK 73190 USA. NR 42 TC 12 Z9 14 U1 0 U2 1 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD AUG PY 2002 VL 43 IS 8 BP 2774 EP 2781 PG 8 WC Ophthalmology SC Ophthalmology GA 577ZZ UT WOS:000177094100038 PM 12147615 ER PT J AU Jeffrey, BG Mitchell, DC Gibson, RA Neuringer, M AF Jeffrey, BG Mitchell, DC Gibson, RA Neuringer, M TI n-3 fatty acid deficiency alters recovery of the rod photoresponse in rhesus monkeys SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article; Proceedings Paper CT Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology CY APR 30-MAY 05, 2000 CL FT LAUDERDALE, FLORIDA SP Assoc Res Vis & Ophthalmol ID RETINOID-BINDING PROTEIN; ERG A-WAVE; DOCOSAHEXAENOIC ACID; RAT RETINA; GUINEA-PIG; HUMAN ELECTRORETINOGRAM; ELECTRICAL RESPONSE; ENZYMATIC-ACTIVITY; DARK-ADAPTATION; OUTER SEGMENTS AB PURPOSE. Docosahexacnoic acid (DHA) is the major poly-unsaturated fatty acid within rod outer segments. Varying the dietary content of DHA or its precursor, alpha-linolenic acid (ALA), can alter retinal DHA levels. The purpose of the present study was to assess rod phototransduction and recovery in rhesus monkeys raised on diets with different DHA and ALA content. METHODS. Adult rhesus monkeys had consumed from birth a diet low in ALA (0.3%) and known to induce an 80% reduction in retinal DHA. They were compared with groups receiving 8% ALA or 0.6% DHA, both of which support normal retinal DHA levels. Rod recovery was assessed with a double-flash protocol using a-wave saturating test flashes. The recovery of rod-isolated ERG a-wave amplitude was analyzed to determine T-c, the time to initiation of rod recovery, and T-50, the time from initiation to 50% of full recovery. Phototransduction was assessed from the fit of a quantitative model to the leading edges of rod-isolated ERG a-waves. ERG a- and b-wave amplitudes and implicit times were also measured. RESULTS. Rod recovery (T50) was delayed by 30% and ERG implicit times by 5% in monkeys in the low ALA group compared with the other groups. There was no significant effect of diet on ERG amplitudes, the time to initiation of rod recovery, or the parameters describing phototransduction. CONCLUSIONS. The results indicate that mechanisms involved in deactivation and rod recovery are selectively altered in monkeys raised on a low-ALA diet whereas, at the flash intensities used, the mechanisms underlying phototransduction remain unaffected. C1 Oregon Hlth & Sci Univ, Oregon Natl Primate Res Ctr, Beaverton, OR 97006 USA. Oregon Hlth & Sci Univ, Casey Eye Inst, Beaverton, OR 97006 USA. Flinders Univ S Australia, Dept Paediat & Child Hlth, Flinders Med Ctr, Adelaide, SA 5001, Australia. NIAAA, Lab Membrane Biochem & Biophys, NIH, Bethesda, MD USA. Child Hlth Res Inst, Childrens Nutr Res Ctr, Adelaide, SA, Australia. RP Neuringer, M (reprint author), Oregon Hlth & Sci Univ, Oregon Natl Primate Res Ctr, 505 NW 185th Ave, Beaverton, OR 97006 USA. RI Gibson, Robert/E-7546-2012 OI Gibson, Robert/0000-0002-8750-525X FU NCRR NIH HHS [RR00163]; NIDDK NIH HHS [DK29930] NR 66 TC 41 Z9 41 U1 1 U2 4 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD AUG PY 2002 VL 43 IS 8 BP 2806 EP 2814 PG 9 WC Ophthalmology SC Ophthalmology GA 577ZZ UT WOS:000177094100042 PM 12147619 ER PT J AU Honda, T Tamura, G Waki, T Jin, Z Sato, K Motoyama, T Kawata, S Kimura, W Nishizuka, S Murakami, Y AF Honda, T Tamura, G Waki, T Jin, Z Sato, K Motoyama, T Kawata, S Kimura, W Nishizuka, S Murakami, Y TI Hypermethylation of the TSLC1 gene promoter in primary gastric cancers and gastric cancer cell lines SO JAPANESE JOURNAL OF CANCER RESEARCH LA English DT Article DE gastric cancer; TSLC1; hypermethylation ID BREAST-CANCER; HETEROZYGOSITY; ADENOCARCINOMA; REGIONS; STOMACH; CHROMOSOME-11Q; IDENTIFICATION; METHYLATION; REFINEMENT AB The TSLC1 (tumor suppressor in lung cancer-1) gene is a novel tumor suppressor gene on chromosomal region 11q23.2, and is frequently inactivated by concordant promoter hypermethylation and loss of heterozygosity (LOH) in non-small cell lung cancer (NSCLC). Because LOH on 11q has also been observed frequently in other human neoplasms including gastric cancer, we investigated the promoter methylation status of TSLC1 in 10 gastric cancer cell lines and 97 primary gastric cancers, as well as the corresponding non-cancerous gastric tissues, by bisulfite-SSCP analysis followed by direct sequencing. Allelic status of the TSLC1 gene was also investigated in these cell lines and primary gastric cancers. The TSLC1 promoter was methylated in two gastric cancer cell lines, KATO-III and ECC10, and in 15 out of 97 (16%) primary gastric cancers. It was not methylated in non-cancerous gastric tissues, suggesting that this hypermethylation is a cancer-specific alteration. KATO-III and ECC10 cells retained two alleles of TSLC1, both of which showed hypermethylation, associated with complete loss of gene expression. Most of the primary gastric cancers with promoter methylation also retained heterozygosity at the TSLC1 locus on 11q23.2. These data indicate that bi-allelic hypermethylation of the TSLC1 promoter and resulting gene silencing occur in a subset of primary gastric cancers. C1 Yamagata Univ, Sch Med, Dept Pathol, Yamagata 9909585, Japan. Yamagata Univ, Sch Med, Dept Med, Yamagata 9909585, Japan. Yamagata Univ, Sch Med, Dept Surg, Yamagata 9909585, Japan. NCI, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA. Natl Canc Ctr, Res Inst, Tumor Suppress & Funct Genomics Project, Chuo Ku, Tokyo 1040045, Japan. RP Tamura, G (reprint author), Yamagata Univ, Sch Med, Dept Pathol, 2-2-2 Iida Nishi, Yamagata 9909585, Japan. NR 13 TC 50 Z9 55 U1 0 U2 0 PU BUSINESS CENTER ACADEMIC SOCIETIES JAPAN PI TOKYO PA 5-16-9 HONKOMAGOME, BUNKYO-KU, TOKYO, 113-8633, JAPAN SN 0910-5050 J9 JPN J CANCER RES JI Jpn. J. Cancer Res. PD AUG PY 2002 VL 93 IS 8 BP 857 EP 860 PG 4 WC Oncology SC Oncology GA 589DP UT WOS:000177743200002 PM 12716461 ER PT J AU Swanson, JM Arnold, LE Vitiello, B Abikoff, HB Wells, KC Pelham, WE March, JS Hinshaw, SP Hoza, B Epstein, JN Elliott, GR Greenhill, LL Hechtman, L Jensen, PS Kraemer, HC Kotkin, R Molina, B Newcorn, JH Owens, EB Severe, J Hoagwood, K Simpson, S Wigal, T Hanley, T AF Swanson, JM Arnold, LE Vitiello, B Abikoff, HB Wells, KC Pelham, WE March, JS Hinshaw, SP Hoza, B Epstein, JN Elliott, GR Greenhill, LL Hechtman, L Jensen, PS Kraemer, HC Kotkin, R Molina, B Newcorn, JH Owens, EB Severe, J Hoagwood, K Simpson, S Wigal, T Hanley, T CA MTA Grp TI Response to commentary on the multimodal treatment study of ADHD (MTA): Mining the meaning of the MTA SO JOURNAL OF ABNORMAL CHILD PSYCHOLOGY LA English DT Article DE MTA; ADHD; behavioral treatment; multimodal treatment; core deficit ID DEFICIT HYPERACTIVITY DISORDER; LONGITUDINAL PSYCHIATRIC DATA; CHILDREN; PROGRAM; DESIGN; ISSUES AB In the December 2000 issue of the Journal of Abnormal Child Psychology, we published a set of papers presenting secondary analyses of the Multimodal Treatment Study of ADHD (MTA), and R. A. Barkley (2000) provided a commentary. A critique of the design of the study (MTA Cooperative Group, 1999) was presented based on a theoretical perspective of a "behavioral inhibition" deficit that has been hypothesized as the core deficit of ADHD (R. A. Barkley, 1997). The commentary questioned the design and analysis of the MTA in terms of (1) the empirical criteria for selection of components of behavioral (Beh) intervention, (2) the effectiveness of the Beh intervention, (3) the methods for analyses at the group and individual level, (4) implications of the MTA findings for clinical practice, (5) the role of genetics in response to treatment, and (6) the lack of a nontreatment control group. In this response, we relate the content of the papers to the commentary, (1) by reviewing the selection criteria for the Beh treatment, as outlined by K. C. Wells, W. E. Pelham, et al. (2000), (2) by addressing the myth that the MTA Beh treatment was ineffective (Pelham, 1999), (3) by describing the use of analyses at the level of the individual participant, as presented by J. S. March et al. (2000) and W. E. Pelham et al. (2000) as well as elsewhere by J. M. Swanson et al. (2001) and C. K. Conners et al. (2001), (4) by relating some of the suggestions from the secondary analyses about clinically relevant factors such as comorbidity (as presented by J. S. March et al., 2000) and family and parental characteristics (as presented by B. Hoza et al., 2000, S. P. Hinshaw et al., 2000, and K. C. Wells, J. N. Epstein, et al., 2000), (5) by discussing the statistical concept of heritability and the lack of a significant difference in the presence of ADHD symptoms in parents of the MTA families compared to parents in the classmate-control families (as presented by J. N. Epstein, et al., 2000), and (6) by acknowledging that an ethically necessary weakness of the MTA design is that it did not include a no-treatment control group. We discuss the use of secondary analyses to suggest how, when, and for what subgroups effectiveness of the Beh treatment may have been manifested. Finally, we invite others to use the large and rich data set that will soon be available in the public domain, to perform secondary analyses to mine the meaning of the MTA and to evaluate theories of ADHD and response to treatments. C1 Univ Calif Irvine, Dept Pediat, Child Dev Ctr, Irvine, CA 92612 USA. Ohio State Univ, Dept Psychiat, Columbus, OH 43210 USA. NIMH, Rockville, MD 20857 USA. NYU, Sch Med, Dept Psychiat, New York, NY USA. Duke Univ, Med Ctr, Dept Psychiat, Durham, NC 27710 USA. SUNY Buffalo, Dept Psychol, Buffalo, NY 14260 USA. Univ Calif Berkeley, Dept Psychol, Berkeley, CA 94720 USA. Purdue Univ, Dept Psychol, W Lafayette, IN 47907 USA. Stanford Univ, Dept Psychiat, Palo Alto, CA 94304 USA. Columbia Univ, New York State Psychiat Inst, New York, NY USA. McGill Univ, Montreal Childrens Hosp, Dept Psychiat, Montreal, PQ H3H 1P3, Canada. Univ Pittsburgh, Dept Psychiat, Pittsburgh, PA USA. Mt Sinai Med Ctr, Dept Psychiat, New York, NY 10029 USA. US Dept Educ, Off Special Educ Projects, Washington, DC USA. RP Swanson, JM (reprint author), Univ Calif Irvine, Dept Pediat, Child Dev Ctr, 19722 MacArthur Blvd, Irvine, CA 92612 USA. OI Jensen, Peter/0000-0003-2387-0650; Newcorn, Jeffrey /0000-0001-8993-9337 FU NIMH NIH HHS [MH50467, U01 MH50440, U01 MH50447, U01 MH50453, U01 MH50454, U01 MH50461, U01 MH50467, U01-MH50453, U01-MH50461] NR 29 TC 13 Z9 14 U1 2 U2 5 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0091-0627 J9 J ABNORM CHILD PSYCH JI J. Abnorm. Child Psychol. PD AUG PY 2002 VL 30 IS 4 BP 327 EP 332 AR UNSP pp502-jacp-374356 PG 6 WC Psychology, Clinical; Psychology, Developmental SC Psychology GA 557TE UT WOS:000175923600002 PM 12108764 ER PT J AU Cohen, SG Zelaya-Quesada, M AF Cohen, SG Zelaya-Quesada, M TI Portier, Richet, and the discovery of anaphylaxis: A centennial SO JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY LA English DT Article C1 NIAID, Bethesda, MD 20892 USA. NIH, Natl Lib Med, Bethesda, MD 20892 USA. RP Cohen, SG (reprint author), NIAID, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 33 TC 15 Z9 18 U1 0 U2 2 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0091-6749 J9 J ALLERGY CLIN IMMUN JI J. Allergy Clin. Immunol. PD AUG PY 2002 VL 110 IS 2 BP 331 EP 336 AR UNSP 1/54/126565 PG 6 WC Allergy; Immunology SC Allergy; Immunology GA 585DY UT WOS:000177509800025 PM 12170279 ER PT J AU Barkmeier, JM Bielamowicz, S Takeda, N Ludlow, CL AF Barkmeier, JM Bielamowicz, S Takeda, N Ludlow, CL TI Laryngeal activity during upright vs. supine swallowing SO JOURNAL OF APPLIED PHYSIOLOGY LA English DT Article DE central pattern generator; larynx ID PHARYNGEAL CONSTRICTOR; VOLUME AB Previous investigations of human pharyngeal muscle activation patterns during swallowing found a relatively invariant muscle activation onset sequence in the upright position. However, different gravitational forces influence a liquid bolus when supine and could modify the central timing control of laryngeal airway protection during swallowing. The purpose of this study was to determine whether laryngeal muscle onset timing during swallowing differed between the supine and upright positions. Nine subjects performed six swallowing trials with a 2-ml water bolus in each position. Simultaneous electromyographic recordings were obtained from the submental complex (SMC) and the right and left thyroarytenoid (TA) muscles. Regardless of body position, the timing, amplitude, and duration of the TA muscles did not vary relative to the SMC. Therefore, the sequence of TA muscle activation relative to the SMC during swallowing appeared unaffected by gravitational influences. C1 NINDS, Laryngeal & Speech Sect, Bethesda, MD 20892 USA. RP Barkmeier, JM (reprint author), Univ Arizona, Dept Speech & Hearing Sci, POB 210071, Tucson, AZ 85710 USA. OI Ludlow, Christy/0000-0002-2015-6171 NR 22 TC 18 Z9 18 U1 0 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 8750-7587 J9 J APPL PHYSIOL JI J. Appl. Physiol. PD AUG PY 2002 VL 93 IS 2 BP 740 EP 745 DI 10.1152/japplphysiol.00380.2001 PG 6 WC Physiology; Sport Sciences SC Physiology; Sport Sciences GA 575GY UT WOS:000176939600043 PM 12133886 ER PT J AU Miyamoto, T Hasuike, S AF Miyamoto, T Hasuike, S TI Isolation and expression analysis of the human RNH2 gene encoding ribonuclease inhibitor 2 SO JOURNAL OF ASSISTED REPRODUCTION AND GENETICS LA English DT Article DE RNH2; testis; spermatogenesis ID LEUCINE-RICH REPEATS; ANGIOGENIN INHIBITOR; PROTEINS AB Ribonuclease inhibitor 1 (RI-1) is a cytoplasmic protein that inhibits a variety of pancreatic-type mammalian Rnases by forming a very tight, reversible 1:1 complex. Recently a novel gene encoding ribonuclease inhibitor 2, Rnh2 has been isolated in mouse. The expression pattern of the mouse Rnh2 is specific to testis, especially in the spermatogonia. Then it has been suggested that the mouse Rnh2 gene may play critical roles in mouse spermatogenesis. In this study, we isolated the human RNH2 cDNA and analyzed its expression pattern. The human RNH2 is also expressed limited to testis, and then it is suggested that the human Rnh2 gene may also play critical roles in human spermatogenesis. C1 NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. RP Miyamoto, T (reprint author), NICHHD, Lab Mammalian Genes & Dev, NIH, Bldg 6B Room 2B211,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 9 TC 4 Z9 4 U1 0 U2 0 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 1058-0468 J9 J ASSIST REPROD GEN JI J. Assist. Reprod. Genet. PD AUG PY 2002 VL 19 IS 8 BP 394 EP 397 AR UNSP pp514-jarg-374644 DI 10.1023/A:1016302706156 PG 4 WC Genetics & Heredity; Obstetrics & Gynecology; Reproductive Biology SC Genetics & Heredity; Obstetrics & Gynecology; Reproductive Biology GA 573CY UT WOS:000176812800007 PM 12182447 ER PT J AU Kim, AD Baker, AS Dunaway-Mariano, D Metcalf, WW Wanner, BL Martin, BM AF Kim, AD Baker, AS Dunaway-Mariano, D Metcalf, WW Wanner, BL Martin, BM TI The 2-aminoethylphosphonate-specific transaminase of the 2-aminoethylphosphonate degradation pathway SO JOURNAL OF BACTERIOLOGY LA English DT Article ID BOND-CLEAVING ENZYME; PHOSPHONOACETALDEHYDE HYDROLASE; PSEUDOMONAS-AERUGINOSA; ASPARTATE-AMINOTRANSFERASE; ESCHERICHIA-COLI; PHOSPHORUS BOND; NEW-GENERATION; PURIFICATION; SEQUENCE; PHOSPHONATASE AB The 2-aminoethylphosphonate transaminase (AEPT; the phnW gene product) of the Salmonella enterica serovar Typhimurium 2-aminoethylphosphonate (AEP) degradation pathway catalyzes the reversible reaction of AEP and pyruvate to form phosphonoacetaldehyde (P-Ald) and L-alanine (L-Ala). Here, we describe the purification and characterization of recombinant AEPT. pH rate profiles (log V-m and log V-m/K-m versus pH) revealed a pH optimum of 8.5. At pH 8.5, K-eq is equal to 0.5 and the k(cat) values of the forward and reverse reactions are 7 and 9 s(-1), respectively. The K-m for AEP is 1.11 +/- 0.03 mM; for pyruvate it is 0.15 +/- 0.02 mM, for P-Ald it is 0.09 +/- 0.01 mM, and for L-Ala it is 1.4 +/- 0.03 mM. Substrate specificity tests revealed a high degree of discrimination, indicating a singular physiological role for the transaminase in AEP degradation. The 40-kDa subunit of the homodimeric enzyme is homologous to other members of the pyridoxalphosphate-dependent amino acid transaminase superfamily. Catalytic residues conserved within well-characterized members are also conserved within the seven known AEPT sequences. Site-directed mutagenesis demonstrated the importance of three selected residues (Asp168, Lys194, and Arg340) in AEPT catalysis. C1 Univ New Mexico, Dept Chem, Albuquerque, NM 87131 USA. Purdue Univ, Dept Sci Biol, W Lafayette, IN USA. Natl Inst Mental Hlth, Bethesda, MD USA. RP Dunaway-Mariano, D (reprint author), Univ New Mexico, Dept Chem, Albuquerque, NM 87131 USA. FU NIGMS NIH HHS [GM28688, GM57695, R01 GM028688] NR 36 TC 28 Z9 29 U1 4 U2 14 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD AUG PY 2002 VL 184 IS 15 BP 4134 EP 4140 DI 10.1128/JB.184.15.4134-4140.2002 PG 7 WC Microbiology SC Microbiology GA 573VY UT WOS:000176853500011 PM 12107130 ER PT J AU Gawel, D Maliszewska-Tkaczyk, M Jonczyk, P Schaaper, RM Fijalkowska, IJ AF Gawel, D Maliszewska-Tkaczyk, M Jonczyk, P Schaaper, RM Fijalkowska, IJ TI Lack of strand bias in UV-induced mutagenesis in Escherichia coli SO JOURNAL OF BACTERIOLOGY LA English DT Article; Proceedings Paper CT 9th International Conference on Microbial Genomes CY 2000 CL LAKE ARROWHEAD, CALIFORNIA ID SOS-INDUCED MUTAGENESIS; DNA-POLYMERASE-V; POL-IV; UNTARGETED MUTAGENESIS; MUTATOR ACTIVITY; RECA PROTEIN; DINB GENE; LESION; UMUC; REPLICATION AB We have investigated whether UV-induced mutations are created with equal efficiency on the leading and lagging strands of DNA replication. We employed an assay system that permits measurement of mutagenesis in the lacZ gene in pairs of near-identical strains. Within each pair, the strains differ only in the orientation of the lacZ gene with respect to the origin of DNA replication. Depending on this orientation, any lacZ target sequence will be replicated in one orientation as a leading strand and as a lagging strand in the other orientation. In contrast to previous results obtained for mutations resulting from spontaneous replication errors or mutations resulting from the spontaneous SOS mutator effect, measurements of UV-induced mutagenesis in uvrA strains fail to show significant differences between the two target orientations. These data suggest that SOS-mediated mutagenic translesion synthesis on the Escherichia coli chromosome may occur with equal or similar probability on leading and lagging strands. C1 Polish Acad Sci, Inst Biochem & Biophys, PL-02106 Warsaw, Poland. NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. RP Fijalkowska, IJ (reprint author), Polish Acad Sci, Inst Biochem & Biophys, Pawinskiego 5A, PL-02106 Warsaw, Poland. RI Fijalkowska, Iwona/I-7796-2016 NR 41 TC 12 Z9 12 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD AUG PY 2002 VL 184 IS 16 BP 4449 EP 4454 DI 10.1128/JB.184.16.4449-4454.2002 PG 6 WC Microbiology SC Microbiology GA 577KF UT WOS:000177059500015 PM 12142415 ER PT J AU Brown, L Gentry, D Elliott, T Cashel, M AF Brown, L Gentry, D Elliott, T Cashel, M TI DksA affects ppGpp induction of RpoS at a translational level SO JOURNAL OF BACTERIOLOGY LA English DT Article; Proceedings Paper CT 9th International Conference on Microbial Genomes CY 2000 CL LAKE ARROWHEAD, CALIFORNIA ID RESPONSE REGULATOR RSSB; SIGMA-FACTOR SIGMA(S); ESCHERICHIA-COLI; SALMONELLA-TYPHIMURIUM; RNA-POLYMERASE; GENE-EXPRESSION; GUANOSINE TETRAPHOSPHATE; MULTICOPY SUPPRESSORS; RECOGNITION FACTOR; UPSTREAM REGION AB The RpoS sigma factor (also called sigma(S) or sigma(38)) is known to regulate at least 50 genes in response to environmental sources of stress or during entry into stationary phase. Regulation of RpoS abundance and activity is complex, with many factors participating at multiple levels. One factor is the nutritional stress signal ppGpp. The absence of ppGpp blocks or delays the induction of rpoS during entry into stationary phase. Artificially inducing ppGpp, without starvation, is known to induce rpoS during the log phase 25- to 50-fold. Induction of ppGpp is found to have only minor effects on rpoS transcript abundance or on RpoS protein stability; instead, the efficiency of rpoS mRNA translation is increased by ppGpp as judged by both RpoS pulse-labeling and promoter-independent effects on lacZ fusions. DksA is found to affect RpoS abundance in a manner related to ppGpp. Deleting dksA blocks rpoS induction by ppGpp. Overproduction of DksA induces rpoS but not ppGpp. Deleting dksA neither alters regulation of ppGpp in response to amino acid starvation nor nullifies the inhibitory effects of ppGpp on stable RNA synthesis. Although this suggests that dksA is epistatic to ppGpp, inducing ppGpp does not induce DksA. A dksA deletion does display a subset of the same multiple-amino-acid requirements found for ppGpp(0) mutants, but overproducing DksA does not satisfy ppGpp(0) requirements. Sequenced spontaneous extragenic suppressors of dksA polyauxotrophy are frequently the same T563P rpoB allele that suppresses a ppGpp(0) phenotype. We propose that DksA functions downstream of ppGpp but indirectly regulates rpoS induction. C1 NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. W Virginia Univ, Hlth Sci Ctr, Dept Microbiol Immunol & Cell Biol, Morgantown, WV 26506 USA. RP Cashel, M (reprint author), NICHHD, Mol Genet Lab, NIH, Bldg 6B,Room 3B-314, Bethesda, MD 20892 USA. NR 57 TC 106 Z9 108 U1 1 U2 7 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD AUG PY 2002 VL 184 IS 16 BP 4455 EP 4465 DI 10.1128/JB.184.16.4455-4465.2002 PG 11 WC Microbiology SC Microbiology GA 577KF UT WOS:000177059500016 PM 12142416 ER PT J AU Hu, JX Mora, S Colussi, G Proverbio, MC Jones, KA Bolzoni, L De Ferrari, ME Civati, G Spiegel, AM AF Hu, JX Mora, S Colussi, G Proverbio, MC Jones, KA Bolzoni, L De Ferrari, ME Civati, G Spiegel, AM TI Autosomal dominant hypocalcemia caused by a novel mutation in the loop 2 region of the human calcium receptor extracellular domain SO JOURNAL OF BONE AND MINERAL RESEARCH LA English DT Article DE autosomal dominant hypocalcemia; hypoparathyroidism; human calcium receptor; mutation; extracellular domain ID HUMAN CA2+ RECEPTOR; FAMILIAL HYPOCALCIURIC HYPERCALCEMIA; NEONATAL SEVERE HYPERPARATHYROIDISM; CELL-SURFACE EXPRESSION; SENSING RECEPTOR; CA2+-SENSING RECEPTOR; SPORADIC HYPOPARATHYROIDISM; ACTIVATING MUTATIONS; SIGNAL-TRANSDUCTION; CHIMERIC RECEPTORS AB We report a novel missense mutation N124K in the extracellular calcium receptor (CaR) identified in two related subjects with the phenotypic features of autosomal dominant hypocalcemia (ADH). Expression of the N124K mutant receptor created by site-directed mutagenesis and transfected into HEK-293 cells was comparable with that of the wild-type (WT) receptor and two other mutant receptors N118K and L125P identified in subjects with ADH. Functional characterization by the extracellular Ca2+ ion ([Ca2+](0))-stimulated phosphoinositide (PI) hydrolysis in transfected HEK-293 cells showed that the N124K mutant receptor was left-shifted in Ca2+ sensitivity. This biochemical gain-of-function is comparable with that seen in other missense mutations of the CaR identified in subjects with ADH. We tested a series of missense substitutions (R, Q, E, and G) in addition to K for N-124 and found that only the N124K mutation and to a much lesser extent N124R caused a left shift in Ca2+ sensitivity. Thus, a specific substitution, not merely a mutation of the N-124 residue, is required for receptor activation. The N124K mutation is one of eight naturally occurring mutations in subjects with ADH identified in a short segment A(116)-C-129 of the CaR extracellular domain (ECD). We present a hypothesis to explain receptor activation by mutations in this region based on the recently described three-dimensional structure of the related metabotropic glutamate type 1 receptor (mGluR1). C1 NIDOCD, Mol Pathophysiol Sect, NIH, Bethesda, MD 20892 USA. Univ Milan, IRCCS HS Raffaele, Lab Pediat Endocrinol, Milan, Italy. Osped Niguarda Ca Granda, Renal Unit, Milan, Italy. RP Hu, JX (reprint author), NIDOCD, Mol Pathophysiol Sect, NIH, Bldg 10,Rm 8C-101,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Mora, Sefano/B-2143-2013 NR 26 TC 19 Z9 19 U1 0 U2 2 PU AMER SOC BONE & MINERAL RES PI WASHINGTON PA 2025 M ST, N W, STE 800, WASHINGTON, DC 20036-3309 USA SN 0884-0431 J9 J BONE MINER RES JI J. Bone Miner. Res. PD AUG PY 2002 VL 17 IS 8 BP 1461 EP 1469 DI 10.1359/jbmr.2002.17.8.1461 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 577EQ UT WOS:000177048900014 PM 12162500 ER PT J AU Takesono, A Finkelstein, LD Schwartzberg, PL AF Takesono, A Finkelstein, LD Schwartzberg, PL TI Beyond calcium: new signaling pathways for Tec family kinases SO JOURNAL OF CELL SCIENCE LA English DT Review DE tyrosine kinase; pleckstrin homology domain; phospholipase C-gamma; calcium mobilization; actin cytoskeleton ID BRUTONS TYROSINE KINASE; PLECKSTRIN HOMOLOGY DOMAIN; T-CELL RECEPTOR; X-LINKED AGAMMAGLOBULINEMIA; ALDRICH-SYNDROME PROTEIN; SRC FAMILY; SH3 DOMAIN; PHOSPHATIDYLINOSITOL 3-KINASE; PH-DOMAIN; PHOSPHOINOSITIDE 3-KINASE AB The Tee kinases represent the second largest family of mammalian non-receptor tyrosine kinases and are distinguished by the presence of distinct proline-rich regions and pleckstrin homology domains that are required for proper regulation and activation. Best studied in lymphocyte and mast cells, these kinases are critical for the full activation of phospholipase-C gamma (PLC-gamma) and Ca2+ mobilization downstream of antigen receptors. However, it has become increasingly clear that these kinases are activated downstream of many cell-surface receptors, including receptor tyrosine kinases, cytokine receptors, integrins and G-protein-coupled receptors. Evidence suggests that the Tee kinases influence a wide range of signaling pathways controlling activation of MAP kinases, actin reorganization, transcriptional regulation, cell survival and cellular transformation. Their impact on cellular physiology suggests that the Tee kinases help regulate multiple cellular processes beyond Ca2+ mobilization. C1 NHGRI, NIH, Bethesda, MD 20892 USA. RP Schwartzberg, PL (reprint author), NHGRI, NIH, 49 Convent Dr,49-4A38, Bethesda, MD 20892 USA. NR 116 TC 61 Z9 64 U1 0 U2 1 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD AUG 1 PY 2002 VL 115 IS 15 BP 3039 EP 3048 PG 10 WC Cell Biology SC Cell Biology GA 586CF UT WOS:000177565500002 PM 12118060 ER PT J AU Bulte, JWM Duncan, ID Frank, JA AF Bulte, JWM Duncan, ID Frank, JA TI In vivo magnetic resonance tracking of magnetically labeled cells after transplantation SO JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM LA English DT Review DE magnetic resonance imaging; MR contrast agent; superparamagnetic iron oxide; stem cell; oligodendrocyte progenitor; transplantation ID SUPERPARAMAGNETIC IRON-OXIDE; CENTRAL-NERVOUS-SYSTEM; EMBRYONIC STEM-CELLS; MYELIN-DEFICIENT RAT; SPINAL-CORD; GLIAL-CELLS; T-CELLS; OLIGODENDROCYTE PROGENITORS; POLYAMIDOAMINE DENDRIMERS; EXTENSIVE MYELINATION AB During the last few years, the therapeutic use of stem and progenitor cells as a substitute for malfunctioning endogenous cell populations has received considerable attention. Unlike their current use in animal models, the introduction of therapeutic cells in patients will require techniques, that can monitor their tissue biodistribution noninvasively, Among the different imaging modalities. magnetic resonance (MR) imaging offers both near-cellular (i.e., 25- to 50-mu) resolution and whole-body imaging capability. In order to be visualized, cells must be labeled with an intracellular tracer molecule that can be detected by MR imaging. Methods have now been developed that make it possible to incorporate sufficient amounts of superparamagnetic iron oxide into cells, enabling their detection in vivo using MR imaging. This is illustrated for (neural stem cell-derived) magnetically labeled oligodendroglial progenitors, transplanted in the central nervous system of dysmyelinated rats. Cells can be followed in vivo for at least 6 weeks after transplantation, with a good histopathologic correlation including the formation of myelin. Now that MR tracking of magnetically labeled cells appears feasible, it is anticipated that this technique may ultimately become an important tool for monitoring the efficacy of clinical (stem) cell transplantation protocols. C1 Johns Hopkins Univ, Sch Med, Dept Radiol, Baltimore, MD 21205 USA. NIH, Expt Neuroimaging Sect, Lab Diagnost Radiol Res, Ctr Clin, Bethesda, MD 20892 USA. Univ Wisconsin, Sch Vet Med, Dept Med Sci, Madison, WI 53706 USA. RP Bulte, JWM (reprint author), Johns Hopkins Univ, Sch Med, Dept Radiol, 217 Traylor Bldg,720 Rutland Ave, Baltimore, MD 21205 USA. RI Bulte, Jeff/A-3240-2008 OI Bulte, Jeff/0000-0003-1202-1610 NR 71 TC 221 Z9 243 U1 1 U2 8 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0271-678X J9 J CEREBR BLOOD F MET JI J. Cereb. Blood Flow Metab. PD AUG PY 2002 VL 22 IS 8 BP 899 EP 907 PG 9 WC Endocrinology & Metabolism; Hematology; Neurosciences SC Endocrinology & Metabolism; Hematology; Neurosciences & Neurology GA 579PL UT WOS:000177187000001 PM 12172375 ER PT J AU Bornstein, MH Painter, KM Park, J AF Bornstein, MH Painter, KM Park, J TI Naturalistic language sampling in typically developing children SO JOURNAL OF CHILD LANGUAGE LA English DT Article ID 2ND YEAR; MOTHER; AGE; SPECIFICITY; VOCABULARY; UTTERANCE; VALIDITY; LENGTH; HOME AB This study compared naturalistic samples of three features of language in 30 two-year-olds - total utterances, word roots, and MLU - in the home in three contrasting situations: the child observed playing by her/himself with mother near by, the child and mother observed in direct play interaction, and the child and mother unobserved at a time the mother judged would provide a sample of the child's 'optimal' language. Children produced more utterances and word roots and expressed themselves in longer MLU when in interaction than when playing 'alone', but children's utterances, word roots, and MLU were greatest in the 'optimal' language production situation. Girls used more word roots and spoke in longer MLU (especially in the 'optimal' language situation) than boys. Despite mean level differences, children maintained their rank orders across the three situations in use of word roots and in MLU. These findings have implications for understanding children's language and the representativeness of sampling child language. C1 NICHHD, Bethesda, MD 20892 USA. RP Bornstein, MH (reprint author), NICHHD, Suite 8030,6705 Rockledge Dr, Bethesda, MD 20892 USA. EM Marc_H_Bornstein@nih.gov NR 36 TC 14 Z9 14 U1 2 U2 6 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 32 AVENUE OF THE AMERICAS, NEW YORK, NY 10013-2473 USA SN 0305-0009 J9 J CHILD LANG JI J. Child Lang. PD AUG PY 2002 VL 29 IS 3 BP 687 EP 699 DI 10.1017/S03050090200524X PG 13 WC Psychology, Developmental; Linguistics; Psychology, Experimental SC Psychology; Linguistics GA 579XF UT WOS:000177203400009 PM 12109369 ER PT J AU Finkelstein, R AF Finkelstein, R TI Future research directions: Follow-up to the Neurofibromatosis Workshop, October 17, 2001 SO JOURNAL OF CHILD NEUROLOGY LA English DT Editorial Material AB This is an exciting time for neurofibromatosis research. During the past decade, researchers isolated the genes that cause the two forms of neurofibromatosis, developed increasingly sophisticated animal models, and learned a great deal about the molecular mechanisms that underlie neurofibromatosis I and neurofibromatosis 2. Most importantly, these advances are beginning to suggest potential therapies for these often devastating disorders. These leaps forward were the result not only of the efforts of a talented community of neurofibromatosis researchers but also of the tireless work of advocacy groups working to facilitate and focus research efforts. The neurofibromatosis workshop held in conjunction with the 30th annual meeting of the Child Neurology Society in Victoria, British Columbia, was a remarkable event. The meeting focused on neurofibromatosis 1, the more prevalent form of the disease. It brought a panel of top neurofibromatosis researchers, clinicians, and patient advocates together with an audience of pediatric neurologists. This was a unique opportunity for researchers to update clinicians regarding their recent discoveries and for clinicians to discuss specific problems that arise in treating patients with neurofibromatosis. In the following sections, I will highlight future research directions suggested at the meeting. C1 NINDS, Bethesda, MD 20892 USA. RP Finkelstein, R (reprint author), NINDS, NSC Room 2143,6001 Execut Blvd, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU B C DECKER INC PI HAMILTON PA 20 HUGHSON ST SOUTH, PO BOX 620, L C D 1, HAMILTON, ONTARIO L8N 3K7, CANADA SN 0883-0738 J9 J CHILD NEUROL JI J. Child Neurol. PD AUG PY 2002 VL 17 IS 8 BP 652 EP 653 DI 10.1177/088307380201700819 PG 2 WC Clinical Neurology; Pediatrics SC Neurosciences & Neurology; Pediatrics GA 602BM UT WOS:000178485100019 PM 12403565 ER PT J AU Egan, JM Meneilly, GS Habener, JF Elahi, D AF Egan, JM Meneilly, GS Habener, JF Elahi, D TI Glucagon-like peptide-1 augments insulin-mediated glucose uptake in the obese state SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID DEPENDENT DIABETES-MELLITUS; RAT SKELETAL-MUSCLE; TURNOVER RATES; SENSITIVITY; RECEPTOR; AMIDE; GLP-1(7-36)AMIDE; SECRETION; KINETICS; HORMONE AB The insulinotropic hormone, glucagon-like peptide-1 (GLP-1), is being examined as a potential new agent for treatment in type 2 diabetic patients. Whereas the insulinotropic properties of this peptide are well established, another property of the hormone, an insulinomimetic effect per se, is controversial. In the normal glucose-tolerant lean state, it is difficult to demonstrate an insulinomimetic effect. The current study was conducted to examine whether GLP-1 has insulinomimetic effect in the obese state. Ten obese volunteers (body mass index, 34.6 +/- 0.8 kg/m(2)), whose ages were 32.5 +/- 3.0 yr, participated in two euglycemic clamp studies (n = 20 clamps) for 120 min. Five of the volunteers were females. The initial clamp was performed with a primed (0-10)-constant (10-60) infusion of GLP-1 at a final rate of 1.5 pmol (.) kg(-1) (.) min(-1). At 60 min, the GLP-1 infusion was terminated, and euglycemic was maintained from 60-120 min. After the GLP-1 study, each individual's plasma insulin level was measured. A second study was performed that was identical to the first, with the infusion of regular insulin in place of GLP-1. Insulin infusion rates were regulated in each individual to simulate plasma insulin levels produced during the GLP-1 infusion. The rate of disappearance of glucose was calculated for each subject. Fasting plasma insulin levels were similar between studies. In response to the GLP-1 infusion, with maintenance of plasma glucose level clamped at fasting level, significant increases in plasma insulin occurred in all subjects (P < 0.001). The insulin levels during the insulin infusion study were similar to that induced by GLP-1. The rate of disappearance of glucose (insulin-mediated glucose uptake) progressively increased in response to both the GLP-1 and insulin infusion. However, the rate of disappearance of glucose during the GLP-1 study was significantly higher (P = 0.033) than during the insulin study. We conclude that in insulin-resistant states, GLP-1 has insulinomimetic properties per se. C1 Harvard Univ, Massachusetts Gen Hosp, Sch Med, Geriatr Res Lab,Dept Med, Boston, MA 02114 USA. NIA, Diabet Sect, Clin Invest Lab, NIH, Baltimore, MD 21224 USA. Univ British Columbia, Dept Med, Vancouver, BC V6T 1Z3, Canada. Howard Hughes Med Inst, Mol Endocrinol Lab, Boston, MA 02114 USA. RP Elahi, D (reprint author), Harvard Univ, Massachusetts Gen Hosp, Sch Med, Geriatr Res Lab,Dept Med, Gray Bigelow Sub Basement 0015,55 Fruit St, Boston, MA 02114 USA. EM delahi@partners.org FU NCRR NIH HHS [M01-RR-02719]; NIA NIH HHS [AG-00599] NR 37 TC 79 Z9 82 U1 0 U2 5 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD AUG PY 2002 VL 87 IS 8 BP 3768 EP 3773 DI 10.1210/jc.87.8.3768 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 581HG UT WOS:000177285400039 PM 12161508 ER PT J AU Shattuck, TM Costa, J Bernstein, M Jensen, RT Chung, DC Arnold, A AF Shattuck, TM Costa, J Bernstein, M Jensen, RT Chung, DC Arnold, A TI Mutational analysis of Smad3, a candidate tumor suppressor implicated in TGF-beta and menin pathways, in parathyroid adenomas and enteropancreatic endocrine tumors SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID GROWTH-FACTOR-BETA; COMPARATIVE GENOMIC HYBRIDIZATION; NEOPLASIA TYPE-1 GENE; TARGETED DISRUPTION; COLORECTAL-CANCER; LOCI; CHROMOSOME-16; TUMORIGENESIS; PROLIFERATION; LOCALIZATION AB Based upon molecular allelotyping and comparative genomic hybridization studies, chromosome 15q is the likely location of a tumor suppressor gene important in the pathogeneses of sporadic enteropancreatic endocrine tumors and parathyroid adenomas. Interest has focused on Smad3 as a candidate endocrine tumor suppressor gene because 1) it is localized to 15q and 2) it encodes a TGFbeta signaling molecule that has been identified as a binding partner of the multiple endocrine neoplasm type 1 gene product menin, itself involved in enteropancreatic and parathyroid neoplasia. To determine whether Smad3 plays a primary role in development of these tumors, 20 enteropancreatic tumors and 67 parathyroid adenomas were investigated for loss of heterozygosity at DNA markers surrounding Smad3. Twenty percent of enteropancreatic tumors and 24% of parathyroid adenomas showed loss. All 9 coding exons and intron-exon boundaries of the Smad3 gene were then sequenced in genomic DNA from all 20 enteropancreatic and 25 parathyroid tumors, including every case with loss of heterozygosity. No acquired clonal mutations, insertions, or microdeletions in Smad3 were detected in any tumors. Because inactivating somatic mutation is the hallmark of an authentic tumor suppressor, Smad3 is unlikely to function as a classical tumor suppressor gene in the pathogenesis of sporadic parathyroid or enteropancreatic endocrine tumors., C1 Univ Connecticut, Sch Med, Ctr Mol Med, Farmington, CT 06030 USA. Univ Connecticut, Sch Med, Div Endocrinol & Metab, Farmington, CT 06030 USA. NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Gastrointestinal Unit, Boston, MA 02114 USA. RP Arnold, A (reprint author), Univ Connecticut, Sch Med, Ctr Mol Med, 263 Farmington Ave, Farmington, CT 06030 USA. EM aarnold@nso2.uchc.edu NR 32 TC 30 Z9 30 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD AUG PY 2002 VL 87 IS 8 BP 3911 EP 3914 DI 10.1210/jc.87.8.3911 PG 4 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 581HG UT WOS:000177285400061 PM 12161532 ER PT J AU Yu, ZY Restifo, NP AF Yu, ZY Restifo, NP TI Cancer vaccines: progress reveals new complexities SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID COLONY-STIMULATING FACTOR; I ANTIGEN EXPRESSION; T-CELLS; MELANOMA-CELLS; METASTATIC MELANOMA; ANTITUMOR IMMUNITY; TUMOR-IMMUNITY; KILLER-CELLS; B2M GENE; IMMUNOTHERAPY C1 NCI, NIH, Bethesda, MD 20892 USA. RP Restifo, NP (reprint author), NCI, NIH, Bldg 10,Room 2B-42, Bethesda, MD 20892 USA. RI Restifo, Nicholas/A-5713-2008; OI Restifo, Nicholas P./0000-0003-4229-4580 FU Intramural NIH HHS [Z99 CA999999, Z01 BC010763-01] NR 56 TC 75 Z9 81 U1 0 U2 2 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD AUG PY 2002 VL 110 IS 3 BP 289 EP 294 DI 10.1172/JCI200216216 PG 6 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 581TH UT WOS:000177308900002 PM 12163445 ER PT J AU Cui, XL Hawari, F Alsaaty, S Lawrence, M Combs, CA Geng, WD Rouhani, FN Miskinis, D Levine, SJ AF Cui, XL Hawari, F Alsaaty, S Lawrence, M Combs, CA Geng, WD Rouhani, FN Miskinis, D Levine, SJ TI Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain shedding SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID TUMOR-NECROSIS-FACTOR; AMINO-ACID-SEQUENCE; FACTOR-ALPHA; CELL-SURFACE; AMINOPEPTIDASE; RECEPTOR; CLONING; METALLOPROTEINASE; DISINTEGRIN; OXYTOCINASE AB Proteolytic cleavage of TNF receptor 1 (TNFR1) generates soluble receptors that regulate TNF bioactivity. We hypothesized that the mechanism of TNFR1 shedding might involve interactions with regulatory ectoproteins. Using a yeast two-hybrid approach, we identified ARTS-I (aminopeptidase regulator of TNFR1 shedding) as a type II integral membrane protein that binds to the TNFR1 extracellular domain. In vivo binding of membrane-associated ARTS-1 to TNFR1 was confirmed by coimmunoprecipitation experiments using human pulmonary epithelial and umbilical vein endothelial cells. A direct relationship exists between membrane-associated ARTS-1 protein levels and concordant changes in TNFR1 shedding. Cells overexpressing ARTS-1 demonstrated increased TNFR1 shedding and decreased membrane-associated TNFR1, while cells expressing antisense ARTS-1 mRNA demonstrated decreased membrane-associated ARTS-1, decreased TNFR1 shedding, and increased membrane-associated TNFR1. ARTS-1 neither bound to TNFR2 nor altered its shedding, suggesting specificity for TNFR1. Although a recombinant ARTS-1 protein demonstrated selective aminopeptidase activity toward nonpolar amino acids, multiple lines of negative evidence suggest that ARTS-1 does not possess TNFR1 sheddase activity. These data indicate that ARTS-1 is a multifunctional ectoprotein capable of binding to and promoting TNFR1 shedding. We propose that formation of a TNFR1-ARTS-1 molecular complex represents a novel mechanism by which TNFR1 shedding is regulated. C1 NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Warren G Magnuson Clin Ctr, Dept Crit Care Med, NIH, Bethesda, MD 20892 USA. NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Levine, SJ (reprint author), NHLBI, Pulm Crit Care Med Branch, NIH, 10 Ctr Dr,Room 6D03,MSC 1590, Bethesda, MD 20892 USA. NR 24 TC 138 Z9 143 U1 0 U2 2 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD AUG PY 2002 VL 110 IS 4 BP 515 EP 526 DI 10.1172/JCI200213847 PG 12 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 585ZL UT WOS:000177557400014 PM 12189246 ER PT J AU Loftin, CD Trivedi, DB Langenbach, R AF Loftin, CD Trivedi, DB Langenbach, R TI Cyclooxygenase-1-selective inhibition prolongs gestation in mice without adverse effects on the ductus arteriosus SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID PRETERM LABOR; ANTENATAL INDOMETHACIN; PLACENTAL-TRANSFER; GENE DISRUPTION; FETAL; RECEPTOR; PROSTAGLANDINS; INFLAMMATION; CONSTRICTION; PARTURITION AB Preterm delivery is the leading cause of neonatal mortality and contributes significantly to infant morbidity. Classical cyclooxygenase (COX) inhibitors, such as indomethacin, which inhibit both COX-1 and COX-2, are effective for delaying premature labor, but their use is limited by serious complications to the fetus and neonate, including adverse effects on the ductus arteriosus (DA). Using isoform-selective inhibitors, we characterized the roles of the COX isoforms in the initiation of labor and the regulation of fetal and neonatal DA closure in mice. Chronic inhibition of COX-2 during pregnancy (gestation days 15-18) significantly increased neonatal mortality by preventing closure of the DA after birth, whereas acute COX-2 inhibition near the end of term (gestation day 18) constricted the fetal DA. In contrast, the inhibition of COX-1 during pregnancy lacked these prenatal and postnatal adverse effects on the DA and effectively delayed the initiation of full-term labor and LPS-induced preterm labor. These findings suggest that premature fetal DA closure or neonatal patent DA observed following indomethacin tocolysis in women may result from the inhibition of COX-2. Therefore, COX-1-selective inhibitors may provide effective treatment to delay preterm labor with fewer adverse effects on fetal or neonatal health than nonselective or COX-2-selective inhibitors. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, NIH, Res Triangle Pk, NC 27709 USA. RP Langenbach, R (reprint author), NIEHS, Lab Environm Carcinogenesis & Mutagenesis, NIH, Mail Drop C4-09,POB 12233, Res Triangle Pk, NC 27709 USA. NR 48 TC 49 Z9 50 U1 0 U2 1 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD AUG PY 2002 VL 110 IS 4 BP 549 EP 557 DI 10.1172/JCI200214924 PG 9 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 585ZL UT WOS:000177557400017 PM 12189249 ER PT J AU Larsen, HH Kovacs, JA Stock, F Vestereng, VH Lundgren, B Fischer, SH Gill, VJ AF Larsen, HH Kovacs, JA Stock, F Vestereng, VH Lundgren, B Fischer, SH Gill, VJ TI Development of a rapid real-time PCR assay for quantitation of Pneumocystis carinii f. sp carinii SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID DIHYDROFOLATE-REDUCTASE GENE; DIHYDROPTEROATE SYNTHASE; DNA AMPLIFICATION; PNEUMONIA AB A method for reliable quantification of Pneumocystis carinii in research models of A carinii pneumonia (PCP) that is more convenient and reproducible than microscopic enumeration of organisms would greatly facilitate investigations of this organism. We developed a rapid quantitative touchdown (QTD) PCR assay for detecting P. carinii f. sp. carinii, the subspecies of P. carinii commonly used in research models of PCP. The assay was based on the single-copy dihydrofolate reductase gene and was able to detect <5 copies of a plasmid standard per tube. It was reproducibly quantitative (r = 0.99) over 6 log values for standards containing &GE;5 copies/tube. Application of the assay to a series of 10-fold dilutions of P. carinii organisms isolated from rat lung demonstrated that it was reproducibly quantitative over 5 log values (r = 0.99). The assay was applied to a recently reported in vitro axenic cultivation system for A carinii and confirmed our microscopy findings that no organism multiplication had occurred during culture. For all cultures analyzed, QTD PCR assays showed a decrease in A carinii DNA that exceeded the expected decrease due to dilution of the inoculum upon transfer. In conclusion, a rapid, sensitive, and reproducible quantitative PCR assay for A carinii f. sp. carinii has been developed and is applicable to in vivo as well as in vitro systems. The assay should prove useful for conducting studies in which quantification of organism burden or growth assessment is critical, such as in vitro antimicrobic susceptibility testing or in vivo immunopathological experiments. C1 Hvidovre Univ Hosp, Dept Infect Dis 144, Copenhagen HIV Programme, DK-2650 Hvidovre, Denmark. NIH, Ctr Clin, Dept Lab Med, Bethesda, MD 20892 USA. NIH, Ctr Clin, Dept Crit Care, Bethesda, MD 20892 USA. Hvidovre Univ Hosp, Dept Clin Microbiol, DK-2650 Hvidovre, Denmark. RP Larsen, HH (reprint author), Hvidovre Univ Hosp, Dept Infect Dis 144, Copenhagen HIV Programme, Kettegaard Alle 30, DK-2650 Hvidovre, Denmark. NR 22 TC 37 Z9 39 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD AUG PY 2002 VL 40 IS 8 BP 2989 EP 2993 DI 10.1128/JCM.40.8.2989-2993.2002 PG 5 WC Microbiology SC Microbiology GA 580VA UT WOS:000177255900046 PM 12149363 ER PT J AU Kiratisin, P Fukuda, CD Wong, A Stock, F Preuss, JC Ediger, L Brahmbhatt, TN Fischer, SH Fedorko, DP Witebsky, FG Gill, VJ AF Kiratisin, P Fukuda, CD Wong, A Stock, F Preuss, JC Ediger, L Brahmbhatt, TN Fischer, SH Fedorko, DP Witebsky, FG Gill, VJ TI Large-scale screening of nasal swabs for Bacillus anthracis: Descriptive summary and discussion of the National Institutes of Health's experience SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article AB In October 2001, a letter containing a large number of anthrax spores was sent through the Brent,rood post office in Washington, D.C., to a United States Senate office on Capitol Hill, resulting in contamination in both places. Several thousand people who worked at these sites were screened for spore exposure by collecting nasal swab samples. We describe here a screening protocol which we, as a level A laboratory, used on very short notice to process a large number of specimens (3,936 swabs) in order to report preliminary results as quickly as possible. Six isolates from our screening met preliminary criteria for Bacillus anthracis identification and were referred for definitive testing. Although none of the isolates was later confirmed to be B. anthracis, we studied these isolates further to define their biochemical characteristics and 16S rRNA sequences. Four of the six isolates were identified as Bacillus megaterium, one was identified as Bacillus cereus, and one was an unidentifiable Bacillus sp. Our results suggest that large-scale nasal-swab screening for potential exposure to anthrax spores, particularly if not done immediately postexposure, may not be very effective for detecting A anthracis but may detect a number of Bacillus spp. that are phenotypically very similar to B. anthracis. C1 NIH, Microbiol Serv, Dept Lab Med, Warren Grant Magnuson Clin Ctr, Bethesda, MD 20892 USA. Natl Naval Med Res Inst, Clin Pathol Serv, Bethesda, MD 20889 USA. RP Kiratisin, P (reprint author), NIH, Microbiol Serv, Dept Lab Med, Warren Grant Magnuson Clin Ctr, Bldg 10,Room 2C385,10 Ctr Dr MSC 1508, Bethesda, MD 20892 USA. EM pkiratisin@mail.cc.nih.gov NR 16 TC 15 Z9 16 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD AUG PY 2002 VL 40 IS 8 BP 3012 EP 3016 DI 10.1128/JCM.40.8.3012-3016.2002 PG 5 WC Microbiology SC Microbiology GA 580VA UT WOS:000177255900050 PM 12149367 ER PT J AU Roth, Y Zangen, A Hallett, M AF Roth, Y Zangen, A Hallett, M TI A coil design for transcranial magnetic stimulation of deep brain regions SO JOURNAL OF CLINICAL NEUROPHYSIOLOGY LA English DT Article DE deep brain stimulation; magnetic coil; tranascranial magnetic stimulation; electrical field; nucleus accumbens; reward ID ELECTRIC-FIELD; ELECTROMAGNETIC INDUCTION; VOLUME CONDUCTOR; REWARD; NERVE; ACTIVATION; CORTEX; ADDICTION; CURRENTS; BEHAVIOR AB Noninvasive magnetic stimulation of the human central nervous system has been used in research and the clinic for several years. However, the coils used previously stimulated mainly the cortical brain regions but could not stimulate deeper brain regions directly. The purpose of the current study was to develop a coil to stimulate deep brain regions. Stimulation of the nucleus accumbens and the nerve fibers connecting the prefrontal cortex with the nucleus accumbens was one major target of the authors' coil design. Numeric simulations of the electrical field induced by several types of coils were performed and accordingly an optimized coil for deep brain stimulation was designed. The electrical field induced by the new coil design was measured in a phantom brain and compared with the double-cone coil. The numeric simulations show that the electrical fields induced by various types of coils are always greater in cortical regions (closer to the coil placement); however, the decrease in electrical field within the brain (as a function of the distance from the coil) is markedly slower for the new coil design. The phantom brain measurements basically confirmed the numeric simulations. The suggested coil is likely to have the ability of deep brain stimulation without the need to increase the intensity to levels that stimulate cortical regions to a much higher extent and possibly cause undesirable side effects. C1 NIDA, NIH, Baltimore, MD 21224 USA. New Adv Technol Ctr, Sheba Med Ctr, Tel Hashomer, Israel. NINDS, NIH, Bethesda, MD 20892 USA. RP Zangen, A (reprint author), NIDA, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 30 TC 106 Z9 109 U1 3 U2 10 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0736-0258 J9 J CLIN NEUROPHYSIOL JI J. Clin. Neurophysiol. PD AUG PY 2002 VL 19 IS 4 BP 361 EP 370 DI 10.1097/00004691-200208000-00008 PG 10 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 611MU UT WOS:000179022100008 PM 12436090 ER PT J AU Solomon, SD Johnson, DM AF Solomon, SD Johnson, DM TI Psychosocial treatment of posttraumatic stress disorder: A practice-friendly review of outcome research SO JOURNAL OF CLINICAL PSYCHOLOGY LA English DT Review DE posttraumatic stress disorder; traumatic stress; treatment; psychotherapy; psychosocial treatment ID METAANALYSIS AB A review of the treatment research indicates that several forms of therapy appear to be useful in reducing the symptoms of posttraumatic stress disorder (PTSD). Strongest support is found for the treatments that combine cognitive and behavioral techniques. Hypnosis, psychodynamic, anxiety management, and group therapies also may produce short-term symptom reduction. Still unknown is whether any approach produces lasting effects. Imaginal exposure to trauma memories and hypnosis are techniques most likely to affect the intrusive symptoms of PTSD, while cognitive and psychodynamic approaches may address better the numbing and avoidance symptoms. Treatment should be tailored to the severity and type of presenting PTSD symptoms, to the type of trauma experience, and to the many likely comorbid diagnoses and adjustment problems, (C) 2002 Wiley Periodicals, Inc. C1 NIH, Off Behav & Social Sci Res, Bethesda, MD 20892 USA. Brown Univ, Providence, RI 02912 USA. RP Solomon, SD (reprint author), NIH, Off Behav & Social Sci Res, 1 Ctr Dr,Room 256, Bethesda, MD 20892 USA. RI Solomon, Scott/I-5789-2013 NR 17 TC 25 Z9 25 U1 1 U2 4 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0021-9762 J9 J CLIN PSYCHOL JI J. Clin. Psychol. PD AUG PY 2002 VL 58 IS 8 BP 947 EP 959 DI 10.1002/jclp.10069 PG 13 WC Psychology, Clinical SC Psychology GA 576QX UT WOS:000177017400007 PM 12115717 ER PT J AU Penzak, SR Hon, YY Lawhorn, WD Shirley, KL Spratlin, V Jann, MW AF Penzak, SR Hon, YY Lawhorn, WD Shirley, KL Spratlin, V Jann, MW TI Influence of ritonavir on olanzapine pharmacokinetics in healthy volunteers SO JOURNAL OF CLINICAL PSYCHOPHARMACOLOGY LA English DT Article ID PLASMA-CONCENTRATIONS; CLINICAL-RESPONSE; HIV; TRIAL AB HIV infection and psychotic illnesses frequently coexist. The atypical antipsychotic olanzapine is metabolized primarily by CYP1A2 and glucuronosyl. transferases, both of which are induced by the HIV protease inhibitor ritonavir. The purpose of this study was to determine the effect of ritonavir on the pharmacokinetics of a single dose of olanzapine. Fourteen healthy volunteers (13 men; age range, 20-28 years) participated in this open-label study. Subjects received olanzapine 10 mg and blood samples were collected over a 120-hour post-dose period. Two weeks later, subjects took ritonavir 300 mg twice daily for 3 days, 400 mg twice daily for 4 days, and 500 mg twice daily for 4 days. The next morning, after 11 days of ritonavir, olanzapine 10 mg was administered and blood sampling was repeated. Plasma samples were analyzed for olanzapine with HPLC. We compared olanzapine noncompartmental pharmacokinetic parameter values before and after ritonavir with a paired Student t test. Ritonavir reduced the area under the plasma concentration-time curve of olanzapine from 501 ng (.) hr/mL (443-582) to 235 ng (.) hr/mL (197-294) (p < 0.001), the half-life from 32 hours (28-36) to 16 hours (14-18) (p = 0.00001), and the peak concentration from 15 ng/mL (13-19) to 9 ng/mL (8-12) (p = 0.002). Olanzapine oral clearance increased from 20 L/hr (18-23) to 43 L/hr (38-51) (p < 0.001) after ritonavir. Ritonavir significantly reduced the systemic exposure of olanzapine in volunteers. Patients receiving this combination may ultimately require higher olanzapine doses to achieve desired therapeutic effects. C1 Mercer Univ, So Sch Pharm, Dept Pharmaceut Sci, Atlanta, GA USA. Mercer Univ, So Sch Pharm, Dept Pharm Practice, Atlanta, GA USA. RP Penzak, SR (reprint author), NIH, Clin Pharmacokinet Lab, Clin Ctr Pharm Dept, Bldg 10,Room 1 N 257, Bethesda, MD 20892 USA. NR 19 TC 27 Z9 28 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0271-0749 J9 J CLIN PSYCHOPHARM JI J. Clin. Psychopharmacol. PD AUG PY 2002 VL 22 IS 4 BP 366 EP 370 DI 10.1097/01.jcp.0000024575.36017.5d PG 5 WC Pharmacology & Pharmacy; Psychiatry SC Pharmacology & Pharmacy; Psychiatry GA 578EH UT WOS:000177104100005 PM 12172335 ER PT J AU Ishai, A AF Ishai, A TI Streams of consciousness SO JOURNAL OF COGNITIVE NEUROSCIENCE LA English DT Article ID CORTEX; REPRESENTATION; OBJECTS C1 NIMH, Lab Brain & Cognit, NIH, Bethesda, MD 20892 USA. RP Ishai, A (reprint author), NIMH, Lab Brain & Cognit, NIH, Bldg 10,Room 4C104, Bethesda, MD 20892 USA. NR 8 TC 0 Z9 0 U1 0 U2 0 PU M I T PRESS PI CAMBRIDGE PA FIVE CAMBRIDGE CENTER, CAMBRIDGE, MA 02142 USA SN 0898-929X J9 J COGNITIVE NEUROSCI JI J. Cogn. Neurosci. PD AUG PY 2002 VL 14 IS 6 BP 832 EP 833 DI 10.1162/089892902760191036 PG 2 WC Neurosciences; Psychology, Experimental SC Neurosciences & Neurology; Psychology GA 583DP UT WOS:000177390900003 PM 12191449 ER PT J AU Dreher, JC Guigon, E Burnod, Y AF Dreher, JC Guigon, E Burnod, Y TI A model of prefrontal cortex dopaminergic modulation during the delayed alternation task SO JOURNAL OF COGNITIVE NEUROSCIENCE LA English DT Article ID SPATIAL WORKING-MEMORY; VENTRAL TEGMENTAL AREA; CORTICAL PYRAMIDAL NEURONS; NUCLEUS-ACCUMBENS; CEREBRAL-CORTEX; D1 RECEPTORS; EXTRACELLULAR DOPAMINE; INHIBITORY INFLUENCE; PRELIMBIC CORTEX; RHESUS-MONKEY AB Working memory performance is modulated by the level of dopamine (DA) D1 receptors stimulation in the prefrontal cortex (PFC). This modulation is exerted at different time scales. Injection of D1 agonists/antagonists exerts a long-lasting influence (several minutes or hours) on PFC pyramidal neurons. In contrast, during performance of a cognitive task, the duration of the postsynaptic effect of phasic DA release is short lasting. The functional relationships of these two time scales of DA modulation remain poorly understood. Here we propose a model that combines these two time scales of DA modulation on a prefrontal neural network. The model links the cellular and behavioral levels during performance of the delayed alternation task. The network, which represents the activity of deep-layer pyramidal neurons with intrinsic neuronal properties, exhibits two stable states of activity that can be switched on and off by excitatory inputs from long-distance cortical areas arriving in superficial layers. These stable states allow PFC neurons to maintain representations during the delay period. The role of an increase of DA receptors stimulation is to restrict inputs arriving on the prefrontal network. The model explains how the level of working memory performance follows an inverted U-shape with an increased stimulation of DA D1 receptors. The model predicts that (1) D1 receptor agonists increase perseverations, (2) D1 antagonists increase distractability, and (3) the duration of the postsynaptic effect of phasic DA release in the PFC is adjusted to the delay period of the task. These results show how the precise duration of the postsynaptic effect of phasic DA release influences behavioral performance during a simple cognitive task. C1 Univ Paris 06, INSERM, U483, F-75252 Paris 05, France. RP Dreher, JC (reprint author), NIMH, Clin Brain Disorders Branch, Room 4C108,MSC 1440, Bethesda, MD 20892 USA. NR 94 TC 30 Z9 32 U1 1 U2 2 PU M I T PRESS PI CAMBRIDGE PA FIVE CAMBRIDGE CENTER, CAMBRIDGE, MA 02142 USA SN 0898-929X J9 J COGNITIVE NEUROSCI JI J. Cogn. Neurosci. PD AUG PY 2002 VL 14 IS 6 BP 853 EP 865 DI 10.1162/089892902760191081 PG 13 WC Neurosciences; Psychology, Experimental SC Neurosciences & Neurology; Psychology GA 583DP UT WOS:000177390900008 PM 12191453 ER PT J AU Goldberg, M Septier, D Rapoport, O Young, M Ameye, L AF Goldberg, M Septier, D Rapoport, O Young, M Ameye, L TI Biglycan is a repressor of amelogenin expression and enamel formation: An emerging hypothesis SO JOURNAL OF DENTAL RESEARCH LA English DT Article DE biglycan; proteoglycan; amelogenin; ameloblasts; odontoblasts ID RAT INCISOR; DENTIN PROTEOGLYCANS; GROWTH-FACTOR; PROTEINS; DECORIN; BONE; PREDENTINE; LOCALIZATION; SULFATE; MINERALIZATION C1 Univ Paris 05, Lab Biol & Physiopathol Craniofaciales, Grp Matrices Extracellulaires & Mineralisat, Fac Chirurg Dent, F-92120 Montrouge, France. NIDCR, CSDB, NIH, Bethesda, MD USA. Nestle Res Ctr, Dept Nutr, CH-1000 Lausanne 26, Switzerland. RP Goldberg, M (reprint author), Univ Paris 05, Lab Biol & Physiopathol Craniofaciales, Grp Matrices Extracellulaires & Mineralisat, Fac Chirurg Dent, 1 Rue Maurice Arnoux, F-92120 Montrouge, France. NR 49 TC 24 Z9 25 U1 1 U2 3 PU INT AMER ASSOC DENTAL RESEARCHI A D R/A A D R PI ALEXANDRIA PA 1619 DUKE ST, ALEXANDRIA, VA 22314-3406 USA SN 0022-0345 J9 J DENT RES JI J. Dent. Res. PD AUG PY 2002 VL 81 IS 8 BP 520 EP 524 PG 5 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 579WL UT WOS:000177201300006 PM 12147740 ER PT J AU Gronthos, S Brahim, J Li, W Fisher, LW Cherman, N Boyde, A DenBesten, P Robey, PG Shi, S AF Gronthos, S Brahim, J Li, W Fisher, LW Cherman, N Boyde, A DenBesten, P Robey, PG Shi, S TI Stem cell properties of human dental pulp stem cells SO JOURNAL OF DENTAL RESEARCH LA English DT Article DE stem cell; odontoblasts; dentin; in vivo transplantation ID MARROW STROMAL CELLS; EXTRACELLULAR-MATRIX PROTEINS; IN-VITRO; GENE-EXPRESSION; GROWTH-FACTORS; BONE; FIBROBLASTS; DIFFERENTIATION; VIVO; TRANSPLANTATION AB In this study, we characterized the self-renewal capability, multi-lineage differentiation capacity, and clonogenic efficiency of human dental pulp stem cells (DPSCs). DPSCs were capable of forming ectopic dentin and associated pulp tissue in vivo. Stromal-like cells were reestablished in culture from primary DPSC transplants and re-transplanted into immunocompromised mice to generate a dentin-pulp-like tissue, demonstrating their self-renewal capability. DPSCs were also found to be capable of differentiating into adipocytes and neural-like cells. The odontogenic potential of 12 individual single-colony-derived DPSC strains was determined. Two-thirds of the single-colony-derived DPSC strains generated abundant ectopic dentin in vivo, while only a limited amount of dentin was detected in the remaining one-third. These results indicate that single-colony-derived DPSC strains differ from each other with respect to their rate of odontogenesis. Taken together, these results demonstrate that DPSCs possess stem-cell-like qualities, including self-renewal capability and multi-lineage differentiation. C1 NIDCR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. NIDCR, Clin Res Core, NIH, Bethesda, MD 20892 USA. Univ Calif San Francisco, Sch Dent, San Francisco, CA 94143 USA. UCL, Dept Anat & Dev Biol, London, England. RP Shi, S (reprint author), NIDCR, Craniofacial & Skeletal Dis Branch, NIH, Bldg 30,Room 228, Bethesda, MD 20892 USA. EM sshi@dir.nidcr.nih.gov RI Robey, Pamela/H-1429-2011 OI Robey, Pamela/0000-0002-5316-5576 NR 24 TC 808 Z9 893 U1 7 U2 107 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 0022-0345 J9 J DENT RES JI J. Dent. Res. PD AUG PY 2002 VL 81 IS 8 BP 531 EP 535 PG 5 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 579WL UT WOS:000177201300008 PM 12147742 ER PT J AU Ladenheim, EE Hampton, LL Whitney, AC White, WO Battey, JF Moran, TH AF Ladenheim, EE Hampton, LL Whitney, AC White, WO Battey, JF Moran, TH TI Disruptions in feeding and body weight control in gastrin-releasing peptide receptor deficient mice SO JOURNAL OF ENDOCRINOLOGY LA English DT Article ID BOMBESIN BINDING-SITES; NEUROMEDIN-B RECEPTOR; ADMINISTERED BOMBESIN; MOLECULAR-CLONING; NERVOUS-SYSTEM; FOOD-INTAKE; SUPPRESSION; ANTAGONIST; SUBTYPES; CELLS AB Bombesin (BN) interacts with two mammalian receptor subtypes termed gastrin-releasing peptide (GRP)preferring (GRP-R) and neuromedin B (NMB)preferring (NMB-R) that may mediate the satiety action of BN. We examined the feeding behavior of mice that were deficient in the GRP-R (GRP-R KO) to assess the overall contribution of this receptor subtype in the feeding actions of BN-related peptides. GRP-R KO mice failed to suppress glucose intake in response to systemically administered BN and GRP(18-27), whereas both peptides elicited a potent reduction of intake in wild-type (WT) mice. Neither GRP-R KO nor WT mice suppressed glucose intake following NNB administration. Unlike the impaired responses to BN-like peptides, the feeding inhibitory action of cholecystokinin was enhanced in GRP-R KO mice. Consistent with behavioral results, GRP-R KO mice also exhibited a reduction in c-Fos immunoreactivity in the nucleus of the solitary tract (NTS) and paraventricular nucleus (PVN) following peripheral administration of BN. An evaluation of meal patterns showed that GRP-R KO mice ate significantly more at each meal than WT mice, although total 24 h food consumption was equivalent. A long-term analysis of body weight revealed a significant elevation in GRP-R KO mice compared with WT littermates beginning at 45 weeks of age. These data suggest that the GRP-R mediates the feeding effects of BN-like peptides and participates in the termination of meals in mice. C1 Johns Hopkins Univ, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21205 USA. NIDOCM, Mol Biol Lab, NIH, Rockville, MD 20850 USA. RP Ladenheim, EE (reprint author), Johns Hopkins Univ, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21205 USA. FU NIDDK NIH HHS [DK046448-09] NR 27 TC 43 Z9 46 U1 0 U2 1 PU SOC ENDOCRINOLOGY PI BRISTOL PA 17/18 THE COURTYARD, WOODLANDS, BRADLEY STOKE, BRISTOL BS32 4NQ, ENGLAND SN 0022-0795 J9 J ENDOCRINOL JI J. Endocrinol. PD AUG PY 2002 VL 174 IS 2 AR UNSP 0022-0795/02/0174-273 DI 10.1677/joe.0.1740273 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 587CV UT WOS:000177623700013 PM 12176666 ER PT J AU Francischetti, IMB Valenzuela, JG Pham, VM Garfield, MK Ribeiro, JMC AF Francischetti, IMB Valenzuela, JG Pham, VM Garfield, MK Ribeiro, JMC TI Toward a catalog for the transcripts and proteins (sialome) from the salivary gland of the malaria vector Anopheles gambiae SO JOURNAL OF EXPERIMENTAL BIOLOGY LA English DT Article DE Anopheles gambiae; mosquito; salivary gland; malaria; proteomics; transcriptome; genomics; blood-sucking insect ID MOSQUITO AEDES-AEGYPTI; PLASMODIUM-FALCIPARUM; PLATELET ACTIVATION; SEQUENCE; PURIFICATION; SPOROZOITES; EXPRESSION; DATABASE; DISEASES; BLOOD AB Hundreds of Anopheles gambiae salivary gland cDNA library clones have been sequenced. A cluster analysis based on sequence similarity at e(-60) grouped the 691 sequences into 251 different clusters that code for proteins with putative secretory, housekeeping, or unknown functions. Among the housekeeping cDNAs, we found sequences predicted to code for novel thioredoxin, tetraspanin, hemopexin, heat shock protein, and TRIO and MBF proteins. Among secreted cDNAs, we found 21 novel A. gambiae salivary sequences including those predicted to encode amylase, calreticulin, selenoprotein, mucin-like protein and 30-kDa allergen, in addition to antigen 5- and D7-related proteins, three novel salivary gland (SG)-like proteins and eight unique putative secreted proteins (Hypothetical Proteins, HP). The electronic version of this paper contains hyperlinks to FASTA-formatted files for each cluster with the best match to the nonredundant (NR) and conserved domain databases (CDD) in addition to CLUSTAL alignments of each cluster. The N terminus of 12 proteins (SG-1, SG-1-like 2, SG-6, HP 8, HP 9-like, 5' nucleotidase, 30-kDa protein, antigen 5- and four D7-related proteins) has been identified by Edman degradation of PVDF-transferred, SDS/PAGE-separated salivary gland proteins. Therefore, we contribute to the generation of a catalog of A. gambiae salivary transcripts and proteins. These data are freely available and will eventually become an invaluable tool to study the role of salivary molecules in parasite-host/vector interactions. C1 NIAID, Parasit Dis Lab, Med Entomol Sect, NIH, Bethesda, MD 20892 USA. Res Technol Branch, Twinbrook, IL USA. RP Ribeiro, JMC (reprint author), NIAID, Parasit Dis Lab, Med Entomol Sect, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Ribeiro, Jose/0000-0002-9107-0818 NR 61 TC 123 Z9 128 U1 0 U2 4 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0022-0949 J9 J EXP BIOL JI J. Exp. Biol. PD AUG PY 2002 VL 205 IS 16 BP 2429 EP 2451 PG 23 WC Biology SC Life Sciences & Biomedicine - Other Topics GA 590ZW UT WOS:000177855700009 PM 12124367 ER PT J AU Pennington, JAT AF Pennington, JAT TI Food composition databases for bioactive food components SO JOURNAL OF FOOD COMPOSITION AND ANALYSIS LA English DT Review DE bioactive components; functional foods; phytochemicals; antioxidants; food composition databases; carotenoids; flavonoids; isoflavones ID NETHERLANDS; FLAVONOIDS; RISK AB A search of the scientific literature and other sources was undertaken to identify and locate data and databases on the presence and amount of bioactive components in foods. The goals were to determine the availability of such data and databases; develop an annotated bibliography; and suggest the next steps to be taken with regard to the development of food composition databases for these components. Databases were identified for carotenoids, six classes of flavonoids, omega-3 fatty acids, and plant sterols. A few bioactive components (e.g., beta-carotene and phytosterol) are currently in some national food composition databases. Some suggested next steps are the development of a directory of information about bioactive component databases (specialized and national databases); the aggregation of data for tannins, allyl sulfides, indoles, lignins, and phenolic acids; the fostering of communication with clinical research community to assist with the design of studies using functional foods; defining the specific bioactive compounds and units of measure needed for food analysts; and the development of detailed, specialized relational databases. Factors that favor the design and maintenance of specialized food composition databases include the limited availability of data for some bioactive components, uncertain quality of the available data, large number of bioactive components, and evolving science about the physiological roles of these components. Specialized databases could capture more detail (e.g., specific descriptions of the foods and samples analyzed, analytical results from individual studies, the number of samples analyzed, mean and median values, and measures of variability) and be useful for developing representative data for food composition databases and for planning and evaluation of clinical and epidemiological research studies on bioactive components. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 NIH, Div Nutr Res Coordinat, Bethesda, MD 20892 USA. RP Pennington, JAT (reprint author), NIH, Div Nutr Res Coordinat, Bethesda, MD 20892 USA. NR 18 TC 53 Z9 55 U1 1 U2 22 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0889-1575 J9 J FOOD COMPOS ANAL JI J. Food Compos. Anal. PD AUG PY 2002 VL 15 IS 4 BP 419 EP 434 DI 10.1006/jfca.2002.1073 PG 16 WC Chemistry, Applied; Food Science & Technology SC Chemistry; Food Science & Technology GA 601GF UT WOS:000178437600010 ER PT J AU O'Malley, AS Lawrence, W Liang, WC Yabroff, R Lynn, J Kerner, J Mandelblatt, J AF O'Malley, AS Lawrence, W Liang, WC Yabroff, R Lynn, J Kerner, J Mandelblatt, J TI Feasibility of mobile cancer screening and prevention SO JOURNAL OF HEALTH CARE FOR THE POOR AND UNDERSERVED LA English DT Review DE underserved; cancer/prevention and control; screening; mobile mammography ID FECAL-OCCULT-BLOOD; PROSTATE-SPECIFIC ANTIGEN; RANDOMIZED CONTROLLED TRIAL; PUBLIC HOSPITAL EMERGENCY; COLORECTAL-CANCER; BREAST-CANCER; CERVICAL-CANCER; OLDER WOMEN; COST-EFFECTIVENESS; NATIONAL SURVEY AB Many areas have high cancer mortality rates and medically underserved populations. This study describes the feasibility (acceptability and costs) of an urban multiphasic (multiple cancers) screening van. Feasibility was evaluated by literature review and informant interviews. Costs were estimated by resource use from urban mobile screening units; decision analysis estimated the costs per cancer detected for breast, cervix, colorectal, and prostate cancer screening. Acceptability of a multiphasic van varied by the informant's perspective. Feasibility and costs were most sensitive to four parameters: age, prior screening history, risk factors, and volume of simultaneous examinations. Subsidized mobile screening facilities may have the potential to reduce cancer morbidity and mortality if they target hard-to-reach underscreened groups, maintain high volume, coordinate with primary care providers, and build on all infrastructure that provides diagnostic and treatment services regardless of ability to pay. It is unclear whether the investment required will translate into a reasonable cost per year of life saved. C1 Georgetown Univ, Med Ctr, Canc Control Progran, Lombardi Canc Ctr, Washington, DC 20007 USA. Georgetown Univ, Med Ctr, Dept Surg, Washington, DC 20007 USA. NCI, Div Canc Control & Pop Sci, NIH, Bethesda, MD 20892 USA. RP O'Malley, AS (reprint author), Georgetown Univ, Med Ctr, Canc Control Progran, Lombardi Canc Ctr, Washington, DC 20007 USA. OI Yabroff, K. Robin/0000-0003-0644-5572 FU AHRQ HHS [HS 08395]; NCI NIH HHS [R01-CA09314, R01-CA72908, R03-CA83338] NR 118 TC 9 Z9 9 U1 6 U2 6 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1049-2089 J9 J HEALTH CARE POOR U JI J. Health Care Poor Underserved PD AUG PY 2002 VL 13 IS 3 BP 298 EP 319 DI 10.1177/10408902013003004 PG 22 WC Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 572FY UT WOS:000176764200004 PM 12152502 ER PT J AU Kittur, DS Wilasrusmee, C Han, WF Xu, RK Burdick, JF Adler, W AF Kittur, DS Wilasrusmee, C Han, WF Xu, RK Burdick, JF Adler, W TI Locally derived cytokines and upregulation of MHC class II genes in allografts SO JOURNAL OF HEART AND LUNG TRANSPLANTATION LA English DT Article ID MAJOR HISTOCOMPATIBILITY COMPLEX; MURINE CARDIAC ALLOGRAFTS; POLYMERASE CHAIN-REACTION; MESSENGER-RNA; REJECTION; EXPRESSION; CELLS; INTERLEUKIN-6; TRANSCRIPTION; RECIPIENTS AB Background: In vitro, various cytokines can modulate the level of expression of major histocompatibility complex (MHC) Class II antigens. Major histocompatibility complex Class II hyperexpression occurs in many immunologic disorders in vivo, but the cytokines that affect this are difficult to analyze because they are produced in small amounts, they act locally, and their mRNAs have short half-lives. Methods: We studied the expression of cytokines known to up-regulate MHC Class II genes in heart allografts in mice from B10.BR donors to B10.D2 recipients by reverse transcription of mRNA and polymerase chain reaction amplification. The I-Abeta(k) gene expression was also studied in the same fully MHC incompatible strain combination. Results: Messenger RNA for interferon (INF)-gamma, interleukin (IL)-4, and tumor necrosis factor (TNF)-alpha, known inducers of MHC Class II expression in vitro, could be detected in allografts either 24 hours before or simultaneously with massive induction of graft specific I-Abeta mRNA. Interleukin-6 mRNA could be detected as early as 1 day after grafting. Conclusions: These data suggest that known cytokine up-regulators of MHC Class II genes, i.e., IFN-gamma, IL-4, and TNF-alpha may contribute to the upregulation of graft-specific MHC Class II antigens during an allograft reaction. Also, IL-6 expression in allografts may result from the stress of the grafting procedure, as it is evident very early after grafting. C1 SUNY Upstate Med Univ, Dept Surg, Syracuse, NY USA. Johns Hopkins Univ, Sch Med, Dept Surg, Baltimore, MD 21205 USA. NIA, Gerontol Res Ctr, Clin Immunol Sect, Baltimore, MD 21224 USA. RP Kittur, DS (reprint author), SUNY Upstate Med Univ, Dept Surg, 750 E Adams St, Syracuse, NY USA. FU NCRR NIH HHS [S07 RR05378]; NIAID NIH HHS [AI15081, AI25550] NR 31 TC 4 Z9 4 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1053-2498 J9 J HEART LUNG TRANSPL JI J. Heart Lung Transplant. PD AUG PY 2002 VL 21 IS 8 BP 882 EP 889 AR PII S1053-2498(02)00407-2 DI 10.1016/S1053-2498(02)00407-2 PG 8 WC Cardiac & Cardiovascular Systems; Respiratory System; Surgery; Transplantation SC Cardiovascular System & Cardiology; Respiratory System; Surgery; Transplantation GA 582LG UT WOS:000177352000007 PM 12163088 ER PT J AU Wolfraim, LA Alkemade, GM Alex, B Sharpe, S Parks, WT Letterio, JJ AF Wolfraim, LA Alkemade, GM Alex, B Sharpe, S Parks, WT Letterio, JJ TI Development and application of fully functional epitope-tagged forms of transforming growth factor-beta SO JOURNAL OF IMMUNOLOGICAL METHODS LA English DT Article DE TGF-beta; epitope tag; biological activity ID TGF-BETA; CRYSTAL-STRUCTURE; GROWTH-FACTOR-BETA-2; SUPERFAMILY; INDUCTION; RECEPTOR; PLASMID AB Administration of transforming growth factor-beta (TGF-beta) has been found to be of therapeutic benefit in various mouse disease models and has potential clinical usefulness. However, the ability to track the distribution of exogenously administered, recombinant forms of these proteins has been restricted by cross-reactivity with endogenous TGF-beta and related TGF-beta isoforms. We describe novel FLAG- and hemagglutinin (HA)-tagged versions of mature TGF-beta1 that retain full biological activity as demonstrated by their ability to inhibit the growth of Mv1Lu epithelial cells, and to induce phosphorylation of the TGF-beta signaling intermediate, smad 2. Intracellular FLAG- and HA-TGF-beta1 can be detected in transfected cells by confocal immunofluorescence microscopy. We also describe sandwich ELISAs designed to specifically detect epitope-tagged TGF-beta and demonstrate the utility of these tagged ligands as probes for TGF-beta receptor expression by flow cytometry. The design of these fully functional epitope-tagged TGF-beta proteins should facilitate studies such as the evaluation of in vivo peptide pharmacodynamics and trafficking of TGF-beta ligand-receptor complexes. (C) 2002 Published by Elsevier Science B.V. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Letterio, JJ (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, Room C629,Bldg 41,41 Lib Dr,MSC 5055, Bethesda, MD 20892 USA. NR 15 TC 10 Z9 10 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-1759 J9 J IMMUNOL METHODS JI J. Immunol. Methods PD AUG 1 PY 2002 VL 266 IS 1-2 BP 7 EP 18 AR PII S0022-1759(02)00090-X DI 10.1016/S0022-1759(02)00090-X PG 12 WC Biochemical Research Methods; Immunology SC Biochemistry & Molecular Biology; Immunology GA 584EC UT WOS:000177453400002 PM 12133618 ER PT J AU Housseau, F Lindsey, KR Oberholtzer, SD Gonzales, MI Boutin, P Moorthy, AK Shankara, S Roberts, BL Topalian, SL AF Housseau, F Lindsey, KR Oberholtzer, SD Gonzales, MI Boutin, P Moorthy, AK Shankara, S Roberts, BL Topalian, SL TI Quantitative real-time RT-PCR as a method for monitoring T lymphocyte reactivity to full-length tyrosinase protein in vaccinated melanoma patients SO JOURNAL OF IMMUNOLOGICAL METHODS LA English DT Article DE tyrosinase; dendritic cells; immune monitoring; adenovirus; quantitative real-time RT-PCR ID TUMOR-INFILTRATING LYMPHOCYTES; MHC CLASS-I; DENDRITIC CELLS; METASTATIC MELANOMA; IMMUNE-RESPONSES; PERIPHERAL-BLOOD; PEPTIDE VACCINE; ANTIGEN; CANCER; IMMUNOTHERAPY AB The major goal of therapeutic cancer vaccine trials is to mediate tumor regression. However, it is critically important to devise in vitro immunological assays that correlate with clinical outcome, for use as surrogate markers of vaccine efficacy. To date, clinical emphasis has been placed on peptide vaccines, but trends towards the use of more complex immunogens such as whole proteins require the development of efficient and sensitive methods for monitoring their immunological effects. In the context of a vaccination trial using full-length tyrosinase (Ty) to immunize patients with metastatic melanoma, a monitoring technique was developed in which autologous dendritic cells (DC) infected with a recombinant adenovirus encoding the Ty protein were used to assess the Ty-specific reactivity of fresh peripheral blood lymphocytes (PBL) collected from patients at different intervals during therapy. Quantitative real-time RT-PCR (qRT-PCR) was used to measure the production of cytokine mRNA by T cells following a 2.5-h incubation with Ty-expressing DC. Two out of ten patients studied demonstrated Ty protein-specific reactivity that increased during and after the period of vaccination. While one of these patients also reacted to an HLA-A1-compatible Ty peptide, the second did not recognize any of the known Ty epitopes, highlighting the importance of this technique for monitoring the effects of complex vaccines. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. Genzyme Corp, Framingham, MA 01701 USA. RP Topalian, SL (reprint author), NCI, Surg Branch, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 32 TC 14 Z9 15 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-1759 J9 J IMMUNOL METHODS JI J. Immunol. Methods PD AUG 1 PY 2002 VL 266 IS 1-2 BP 87 EP 103 AR PII S0022-1759(02)00104-7 DI 10.1016/S0022-1759(02)00104-7 PG 17 WC Biochemical Research Methods; Immunology SC Biochemistry & Molecular Biology; Immunology GA 584EC UT WOS:000177453400009 PM 12133625 ER PT J AU Bitmansour, AD Douek, DC Maino, VC Picker, LJ AF Bitmansour, AD Douek, DC Maino, VC Picker, LJ TI Direct ex vivo analysis of human CD4(+) memory T cell activation requirements at the single clonotype level SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MHC CLASS-II; DENDRITIC CELLS; LYMPHOCYTE-RESPONSES; ANTIGEN PRESENTATION; EFFECTOR FUNCTION; PEPTIDE LIGANDS; FLOW-CYTOMETRY; IN-VIVO; RECEPTOR; THRESHOLDS AB CD4(+) memory T cells continuously integrate signals transmitted through the TCR and costimulatory molecules, only responding when the intensity of such signals exceeds an intrinsic activation threshold. Recent data suggest that these activation thresholds can be regulated independently of TCR specificity, and that threshold tuning may constitute a major mechanism for controlling T cell effector activity. In this work we take advantage of the profound clonotypic hierarchies of the large human CD4(+) T cell response to CMV to study activation thresholds of fresh (unexpanded) memory T cells at the clonotypic level. We identified dominant responses to CMV matrix determinants mediated by single TCRB sequences within particular TCR-Vbeta families. The specific response characteristics of these single, Ag-specific, TCRB-defined clonotypes could be unequivocally determined in fresh PBMC preparations by cytokine flow cytometry with gating on the appropriate Vbeta family. These analyses revealed 1) optimal peptides capable of eliciting specific responses by themselves at doses as low as 2 pg/ml, with each log increase in dose eliciting ever-increasing frequencies of responding cells over a 4- to 5-log range; 2) significant augmentation of response frequencies at all submaximal peptide doses by CD28- and CD49d-mediated costimulation; 3) differential dose response and costimulatory characteristics for IFN-gamma and IL-2 responses; and 4) no association of activation requirements with the CD27-defined CD4(+) T cell memory differentiation pathway. Taken together these data confirm that triggering heterogeneity exists within individual CD4(+) memory T cell clonotypes in vivo and demonstrate that such single clonotypes can manifest qualitatively different functional responses depending on epitope dose and relative levels of costimulation. C1 Oregon Hlth & Sci Univ, Vaccine & Gene Therapy Inst, Portland, OR 97201 USA. Univ Texas, SW Med Ctr, Grad Program Immunol, Dallas, TX 75390 USA. NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. BD Biosci, San Jose, CA 95131 USA. RP Picker, LJ (reprint author), Oregon Hlth & Sci Univ, Vaccine & Gene Therapy Inst, W Campus,505 NW 185th Ave, Beaverton, OR 97006 USA. FU NIAID NIH HHS [R01-AI47606-03] NR 51 TC 53 Z9 53 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD AUG 1 PY 2002 VL 169 IS 3 BP 1207 EP 1218 PG 12 WC Immunology SC Immunology GA 576UV UT WOS:000177025100009 PM 12133941 ER PT J AU Tsujimura, H Nagamura-Inoue, T Tamura, T Ozato, K AF Tsujimura, H Nagamura-Inoue, T Tamura, T Ozato, K TI IFN consensus sequence binding protein/IFN regulatory factor-8 guides bone marrow progenitor cells toward the macrophage lineage SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HEMATOPOIETIC STEM-CELLS; TRANSCRIPTION FACTOR; MYELOID PROGENITORS; INTERFERON-GAMMA; DEFICIENT MICE; FACTOR FAMILY; EXPRESSION; LEUKEMIA; ICSBP; INDUCTION AB IFN consensus sequence binding protein (ICSBP; IFN regulatory factor-8) is a transcription factor of the IFN regulatory factor family. Disruption of this gene results in a leukemia-like disease in mice. To investigate the role of ICSBP in myeloid cell development, lineage marker-negative (Lin(-)) bone marrow progenitor cells were purified from ICSBP+/+ and ICSBP-/- mice and tested for gene expression and colony-forming ability. ICSBP was expressed in Lin- progenitor cells, and its levels were markedly increased by IFN-gamma. The colony-forming potential of ICSBP-/- progenitor cells was grossly abnormal, as they gave rise to a disproportionately high number of granulocyte colonies and many fewer macrophage colonies. IFN-gamma inhibited colony formation, while promoting macrophage maturation in ICSBP+/+ cells. In contrast, the effects of IFN-gamma were completely absent in ICSBP-/- progenitors. By retrovirus transduction we tested whether reintroduction of ICSBP restores a normal colony-forming potential in -/- progenitor cells. The wild-type ICSBP, but not transcriptionally defective mutants, corrected abnormal colony formation by increasing macrophage colonies and decreasing granulocyte colonies. Taken together, ICSBP plays a critical role in myeloid cell development by controlling lineage selection and is indispensable for IFN-gamma-dependent modulation of progenitor cell maturation. C1 NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Ozato, K (reprint author), NICHHD, Lab Mol Growth Regulat, NIH, Bldg 6,Room 2A01, Bethesda, MD 20892 USA. NR 47 TC 61 Z9 62 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD AUG 1 PY 2002 VL 169 IS 3 BP 1261 EP 1269 PG 9 WC Immunology SC Immunology GA 576UV UT WOS:000177025100015 PM 12133947 ER PT J AU Park, MK Amichay, D Love, P Wick, E Liao, F Grinberg, A Rabin, RL Zhang, HWH Gebeyehu, S Wright, TM Iwasaki, A Weng, YM DeMartino, JA Elkins, KL Farber, JM AF Park, MK Amichay, D Love, P Wick, E Liao, F Grinberg, A Rabin, RL Zhang, HWH Gebeyehu, S Wright, TM Iwasaki, A Weng, YM DeMartino, JA Elkins, KL Farber, JM TI The CXC chemokine murine monokine induced by IFN-gamma (CXC chemokine ligand 9) is made by APCs, targets lymphocytes including activated B cells, and supports antibody responses to a bacterial pathogen in vivo SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TULARENSIS STRAIN LVS; T-CELLS; IN-VIVO; DENDRITIC CELLS; RECEPTOR CXCR3; INTERFERON-GAMMA; INTRACELLULAR PATHOGEN; RHEUMATOID-ARTHRITIS; PROTECTIVE IMMUNITY; ACUTE REJECTION AB Monokine induced by IFN-gamma (Mig; CXC chemokine ligand 9) is an IFN-gamma-inducible CXC chemokine that signals through the receptor CXCR3 and is known to function as a chemotactic factor for human T cells, particularly following T cell activation. The mig gene can be induced in multiple cell types and organs, and Mig has been shown to contribute to T cell infiltration into immune/inflammatory reactions in peripheral tissues in mice. We have investigated the expression and activities of Mig and CXCR3 in mouse cells and the role of Mig in models of host defense in mice. Murine (Mu)Mig functioned as a chemotactic factor for resting memory and activated T cells, both CD4(+) and CD8(+), and responsiveness to MuMig correlated with surface expression of MuCXCR3. Using mig(-/-) mice, we found that MuMig was not necessary for survival after infections with a number of intracellular pathogens. Surprisingly, however, we found that mig(-/-) mice showed reductions of 50-75% in Abs produced against the intracellular bacterium Francisella tularensis live vaccine strain. Furthermore, we found that MuMig induced both calcium signals and chemotaxis in activated B cells, and that B cell activation induced expression of MuCXCR3. In addition, IFN-gamma induced the expression of mumig in APCs, including CD8alpha(+) and CD8alpha(-) dendritic cells. Together, our data suggest that Mig and CXCR3 may be important not only to recruit T cells to peripheral inflammatory sites, but also in some cases to maximize interactions among activated T cells, B cells, and dendritic cells within lymphoid organs to provide optimal Immoral responses to pathogens. C1 NIAID, Clin Invest Lab, Inflammat Biol Sect, NIH, Bethesda, MD 20892 USA. NIAID, Immune Cell Interact Unit, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Merck Res Labs, Dept Immunol Res, Rahway, NJ 07065 USA. Pfizer Global R&D, Ann Arbor, MI 48105 USA. US FDA, Ctr Biol Evaluat & Res, Div Bacterial Parasit & Allergen Prod, Bethesda, MD 20892 USA. NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. RP Farber, JM (reprint author), NIAID, Clin Invest Lab, Inflammat Biol Sect, NIH, Bldg 10,Room 11N228,MSC-1888, Bethesda, MD 20892 USA. NR 60 TC 81 Z9 84 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD AUG 1 PY 2002 VL 169 IS 3 BP 1433 EP 1443 PG 11 WC Immunology SC Immunology GA 576UV UT WOS:000177025100037 PM 12133969 ER PT J AU Patera, A Ali, MA Tyring, S Warren, T Hohman, P Yu, E Cohen, JI AF Patera, A Ali, MA Tyring, S Warren, T Hohman, P Yu, E Cohen, JI TI Polymorphisms in the genes for herpesvirus entry SO JOURNAL OF INFECTIOUS DISEASES LA English DT Letter ID MEDIATOR C1 NIH, Clin Invest Lab, Med Virol Sect, Bethesda, MD 20892 USA. Univ Texas, Med Branch, Dept Microbiol & Immunol, Galveston, TX 77550 USA. Westover Hts Clin, Portland, OR USA. RP Cohen, JI (reprint author), NIH, Clin Invest Lab, Med Virol Sect, Bldg 10,Rm 11N228, Bethesda, MD 20892 USA. NR 4 TC 1 Z9 1 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD AUG 1 PY 2002 VL 186 IS 3 BP 444 EP 445 DI 10.1086/341562 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 578HH UT WOS:000177111000023 PM 12134247 ER PT J AU Piguet, V Blauvelt, A AF Piguet, V Blauvelt, A TI Essential roles for dendritic cells in the pathogenesis and potential treatment of HIV disease SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE Langermans cells; cells; CCRS; DC-SIGN; AIDS ID IMMUNODEFICIENCY-VIRUS TYPE-1; EPIDERMAL LANGERHANS CELLS; CLASS-II COMPLEXES; CD4(+) T-CELLS; DC-SIGN; LENTIVIRAL VECTORS; PRIMARY INFECTION; CHEMOKINE-CORECEPTORS; PRODUCTIVE INFECTION; ENDOTHELIAL-CELLS AB During sexual transmission of HIV, virus crosses mucosal epithelium and eventually reaches lymphoid tissue where it establishes a permanent infection. Evidence has accumulated that infection of Langerhans cells, which are resident dendritic cells in pluristratified epithelia, plays a crucial role in the early events of HIV transmission. HIV infection of Langerhans cells is regulated by surface expression of CD4 and CCR5. Thus, topical microbicides that interfere with HIV infection of Langerhans cells represent an attractive strategy for blocking sexual transmission of virus. Capture and uptake of HIV virions is another major pathway by which HIV interacts with dendritic cells. By contrast, this process is mediated by a newly described C-type lectin, DC-SIGN. It is well established that HIV-exposed dendritic cells transmit virus efficiently to cocultured T cells. Indeed, dendritic cell-T cell interaction, critical in the generation of immune responses, is a rich microenvironment for HIV replication both in vitro and in vivo . Dendritic cells that have captured virus via DC-SIGN, and not HIV-infected dendritic cells, probably facilitate most infection of T cells in chronically infected individuals. Therefore, blocking DC-SIGN-mediated capture of HIV represents a potential therapeutic antiviral strategy for HIV disease. Lastly, dendritic cells have been targeted both ex vivo and in vivo to initiate and enhance HIV-specific immunity. Although these approaches are promising for both therapeutic and prophylactic vaccines, much additional work is needed in order to optimize dendritic-cell-based immunization strategies. C1 Univ Hosp Geneva, Dept Dermatol, CH-1211 Geneva, Switzerland. NCI, Dermatol Branch, Bethesda, MD 20892 USA. RP Piguet, V (reprint author), Univ Hosp Geneva, Dept Dermatol, 24 Rue Micheli du Crest, CH-1211 Geneva, Switzerland. NR 79 TC 52 Z9 55 U1 0 U2 0 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD AUG PY 2002 VL 119 IS 2 BP 365 EP 369 DI 10.1046/j.1523-1747.2002.01840.x PG 5 WC Dermatology SC Dermatology GA 586NL UT WOS:000177590000001 PM 12190858 ER PT J AU Kee, SH Jang, SI Ahvazi, B Larsen, M Yamada, KM Steinert, PM AF Kee, SH Jang, SI Ahvazi, B Larsen, M Yamada, KM Steinert, PM TI Cell-cell adhesion and RhoA-mediated actin polymerization are independent phenomena in microtubule disrupted keratinocytes SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE actin; adherens junction; E-cadherin ID PROTEIN-KINASE-C; PHOSPHOLIPASE-D; SIGNAL-TRANSDUCTION; CADHERIN FUNCTION; HUMAN FIBROBLASTS; ADP-RIBOSYLATION; FAMILY PROTEINS; ALPHA-CATENIN; SMALL GTPASES; DNA-SYNTHESIS AB E-cadherin-mediated adherens junction formation and maintenance are thought to involve actin filament rearrangements through the action of small GTPases. Recently, we demonstrated that microtubule disruption in normal human epidermal keratinocytes grown in low calcium media conditions induces cell-cell adhesion by redistribution of endogenous E-cadherin, and it promotes stress fiber formation. This actin rearrangement was apparently mediated by RhoA activation. This model system therefore provides a tool with which to dissect relationships between cell-cell adhesion and Rho-mediated stress fiber formation. In this study, we have demonstrated in normal human epidermal keratinocytes that disruption of actin structures including stress fibers does not interfere with E-cadherin redistribution during microtubule-induced cell-cell adhesion. Moreover, this cell-cell adhesion could not be blocked by RhoA inactivation at the level for inhibition of stress fiber formation. Additionally, in the immortalized HaCaT keratinocyte cell line, which does not undergo cell-cell adhesion after microtubule disruption in low calcium conditions, expression of dominant-active RhoA could induce stress fiber formation without inducing adhesion. On the other hand, a variant of the HaCaT cell line, HC-R1, showed microtubule-disruption-induced cell-cell adhesion without stress fiber formation. Together, our results suggest that, in keratinocytes, the process of cell adhesion can occur independently of RhoA-mediated stress fiber formation. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD USA. RP Steinert, PM (reprint author), NIAMSD, Skin Biol Lab, NIH, Bldg 50,Room 1523, Bethesda, MD 20892 USA. OI Yamada, Kenneth/0000-0003-1512-6805 NR 48 TC 8 Z9 8 U1 0 U2 0 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD AUG PY 2002 VL 119 IS 2 BP 440 EP 448 DI 10.1046/j.1523-1747.2002.01826.x PG 9 WC Dermatology SC Dermatology GA 586NL UT WOS:000177590000011 PM 12190868 ER PT J AU Huang, W Ishii, I Zhang, WY Sonobe, M Kruth, HS AF Huang, W Ishii, I Zhang, WY Sonobe, M Kruth, HS TI PMA activation of macrophages alters macrophage metabolism of aggregated LDL SO JOURNAL OF LIPID RESEARCH LA English DT Article DE acyl-CoA : cholesterol acyltransferase; atherosclerosis; cholesterol; endocytosis; plasmin; low density lipoprotein; surface-connected compartments; phorbol myristate acetate; protein kinase C; trichloroacetic acid; lysosomes; patocytosis ID LOW-DENSITY-LIPOPROTEIN; MONOCYTE-DERIVED MACROPHAGES; SMOOTH-MUSCLE CELLS; PROTEIN-KINASE-C; ATHEROSCLEROTIC LESIONS; CELLULAR CHOLESTEROL; LIPID-ACCUMULATION; CULTURED-CELLS; EFFLUX; RECEPTOR AB Aggregation of LDL may contribute to its retention in atherosclerotic lesions. Previously, we showed that aggregated LDL induces and enters surface-connected compartments (SCCs) in human monocyte-derived macrophages by a process we have named patocytosis. Aggregated LDL was disaggregated and released from SCCs of macrophages when exposed to human lipoprotein-deficient serum. The serum factor that mediated aggregated LDL release and disaggregation was plasmin generated from plasminogen by macrophage urokinase plasminogen activator. We now show that activation of macrophages with PMA inhibits plasmin-mediated release of aggregated LDL from macrophages. With macrophage activation, plasminogen released about 60% less cholesterol and 63% less TCA-insoluble I-125-aggregated LDL than when macrophages were not activated. Electron microscopy showed that PMA did not cause SCCs to close, which could have trapped aggregated LDL within the SCCs and limited protease access to aggregated LDL. Rather, PMA decreased macrophage generation of plasmin by 61%, and stimulated lysosomal degradation of aggregated LDL by more than 2-fold. Degradation was mediated by protein kinase C, shown by the finding that degradation was inhibited by the protein kinase C inhibitor G66976. PMA-stimulated degradation of aggregated LDL was associated with a 3-fold increase in cholesterol esterification, consistent with hydrolysis and re-esterification of aggregated LDL-derived cholesteryl ester.(jlr) In conclusion, macrophage activation with PMA causes more of the aggregated LDL that enters macrophage SCCs to be metabolized by lysosomes. This results in more cholesterol to be stored in macrophages and less aggregated LDL to be available for plasmin-mediated release from macrophage SCCs. C1 NHLBI, Sect Expt Atherosclerosis, NIH, Bethesda, MD 20892 USA. RP Kruth, HS (reprint author), NHLBI, Sect Expt Atherosclerosis, NIH, Bldg 10,Room 5N-113,10 Ctr Dr MSC 1422, Bethesda, MD 20892 USA. RI Huang, Wei/E-3270-2011 NR 36 TC 11 Z9 11 U1 0 U2 0 PU LIPID RESEARCH INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD AUG PY 2002 VL 43 IS 8 BP 1275 EP 1282 DI 10.1194/jlr.M100436-JLR200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 590AL UT WOS:000177795000012 PM 12177171 ER PT J AU Cierpicki, T Zhukov, I Byrd, RA Otlewski, J AF Cierpicki, T Zhukov, I Byrd, RA Otlewski, J TI Hydrogen bonds in human ubiquitin reflected in temperature coefficients of amide protons SO JOURNAL OF MAGNETIC RESONANCE LA English DT Article DE amide proton temperature coefficients; chemical shifts; hydrogen bonds; ubiquitin ID CHEMICAL-SHIFTS; PROTEINS; GRADIENTS AB Analysis of amide proton temperature coefficients (Deltasigma(HN)/DeltaT) in human ubiquitin shows their usefulness in indicating hydrogen bonds. The availability of a very accurate solution structure of ubiquitin enables the precise determination of hydrogen bonds and increases the reliability of the analysis of chemical shift temperature gradients. Values of Deltasigma(HN)/DeltaT more positive than -4.6 ppb/K are very good indicators of hydrogen bonds. Additionally, a weak temperature dependence of non-hydrogen-bonded amides was observed for amide protons that are significantly shifted upfield. We observed that temperature gradients of amide protons involved in short hydrogen bonds are related to donor-acceptor distances. (C) 2002 Elsevier Science (USA). C1 Univ Wroclaw, Inst Biochem & Mol Biol, Lab Prot Engn, PL-50137 Wroclaw, Poland. Polish Acad Sci, Inst Biochem & Biophys, PL-02106 Warsaw, Poland. NCI, Struct Biophys Lab, Frederick, MD 21702 USA. RP Cierpicki, T (reprint author), Univ Wroclaw, Inst Biochem & Mol Biol, Lab Prot Engn, Tamka 2, PL-50137 Wroclaw, Poland. RI Otlewski, Jacek/B-6340-2008; Byrd, R. Andrew/F-8042-2015 OI Byrd, R. Andrew/0000-0003-3625-4232 NR 13 TC 44 Z9 44 U1 2 U2 11 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1090-7807 J9 J MAGN RESON JI J. Magn. Reson. PD AUG PY 2002 VL 157 IS 2 BP 178 EP 180 DI 10.1006/jmre.2002.2597 PG 3 WC Biochemical Research Methods; Physics, Atomic, Molecular & Chemical; Spectroscopy SC Biochemistry & Molecular Biology; Physics; Spectroscopy GA 600EJ UT WOS:000178377400002 PM 12323135 ER PT J AU Ling, B Veazey, RS Penedo, C Xu, K Lifson, JD Marx, PA AF Ling, B Veazey, RS Penedo, C Xu, K Lifson, JD Marx, PA TI Longitudinal follow up of SIVmac pathogenesis in rhesus macaques of Chinese origin: emergence of B cell lymphoma SO JOURNAL OF MEDICAL PRIMATOLOGY LA English DT Article; Proceedings Paper CT 19th Annual Symposium on Nonhuman Primate Models for AIDS CY NOV 07-10, 2001 CL SAN JUAN, PUERTO RICO SP BD Biosci Pharmingen, Glaxo, Res Ctr Minor Inst Program, Univ Puerto Rico, Med Sci Campus, Off Chancellor, Unit Comparat Med, Virol Lab, Dept Microl DE animal experiment; intestinal biopsy; Macaca mulatta; plasma viral load ID SIMIAN IMMUNODEFICIENCY VIRUS; VIRAL LOAD; DISEASE PROGRESSION; ANTIBODY-RESPONSES; TYPE-1 INFECTION; HIV-1 INFECTION; AIDS; TRANSMISSION; EXPRESSION; VACCINE AB Two subspecies of rhesus (Rh) macaques, the Chinese (Ch) and Indian (Ind) subspecies were infected intravenously with 100TCID(50) SIVmac239. CD4+, CD8+ T cells, plasma viral loads, depletion of intestinal lymphocytes with memory phenotype, humoral immune responses and clinical courses were monitored for 600 days. The pathogenesis of SIVmac was also compared with primary human immunodeficiency virus (HIV) infection of humans. Plasma viral loads in Ch Rh were lower in the acute and chronic phases compared with Ind Rh. SIVmac pathogenesis in Ch Rh was closer to virus loads in untreated HIV infected humans. Ch Rh had higher CD4/CD8 ratios, stronger antibody responses and interestingly, less depletion of intestinal memory CCR5+ CD4+ T lymphocytes compared with Ind Rh. One Ch Rh developed B cell origin lymphoma at 570 days post-infection, the first such report in this subspecies. Three of four Ind Rh developed AIDS within 6 months. The findings indicate that Ch Rh are more resistant to SIVmac pathogenesis compared with Ind Rh and that Ch Rh paralleled HIV-1 infections in untreated adult humans. The SIVmac infected Ch Rh subspecies are an acceptable model for HIV/AIDS. C1 Tulane Univ, Tulane Natl Primate Res Ctr, Hlth Sci Ctr, Covington, LA 70433 USA. Rockefeller Univ, Aaron Diamond AIDS Res Ctr, New York, NY 10021 USA. Univ Calif Davis, Vet Genet Lab, Davis, CA 95616 USA. NCI, Frederick Canc Res & Dev Ctr, AIDS Vaccine Program, Frederick, MD USA. RP Marx, PA (reprint author), Tulane Univ, Tulane Natl Primate Res Ctr, Hlth Sci Ctr, 18703 3 Rivers Rd, Covington, LA 70433 USA. FU NCRR NIH HHS [P51RR000164-40]; NIAID NIH HHS [N01-AI65300] NR 31 TC 17 Z9 18 U1 0 U2 1 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0047-2565 J9 J MED PRIMATOL JI J. Med. Primatol. PD AUG PY 2002 VL 31 IS 4-5 BP 154 EP 163 DI 10.1034/j.1600-0684.2002.02001.x PG 10 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 594AT UT WOS:000178027200002 PM 12390537 ER PT J AU Lifson, JD Piatak, M Rossio, JL Bess, J Chertova, E Schneider, D Kiser, R Coalter, V Poore, B Imming, R Desrosiers, RC Henderson, LE Arthur, LO AF Lifson, JD Piatak, M Rossio, JL Bess, J Chertova, E Schneider, D Kiser, R Coalter, V Poore, B Imming, R Desrosiers, RC Henderson, LE Arthur, LO TI Whole inactivated SIV virion vaccines with functional envelope glycoproteins: safety, immunogenicity, and activity against intrarectal challenge SO JOURNAL OF MEDICAL PRIMATOLOGY LA English DT Article; Proceedings Paper CT 19th Annual Symposium on Nonhuman Primate Models for AIDS CY NOV 07-10, 2001 CL SAN JUAN, PUERTO RICO SP BD Biosci Pharmingen, Glaxo, Res Ctr Minor Inst Program, Univ Puerto Rico, Med Sci Campus, Off Chancellor, Unit Comparat Med, Virol Lab, Dept Microl DE SIV; inactivated; vaccine ID SIMIAN IMMUNODEFICIENCY VIRUS; NEUTRALIZATION SENSITIVITY; IMMUNE-RESPONSES; MACAQUES; PROTECTION; PROTEINS; ANTIBODY; CELLS; INFECTIVITY; ANTIGENS AB A novel type of whole inactivated simian immunodeficiency virus (SIV) virion vaccine immunogen with functional envelope glycoproteins was evaluated, without adjuvant, in rhesus macaques. Immunogens included purified inactivated virions of SIVmac239, a designed mutant of SIVmac239 with gp120 carbohydrate attachment sites deleted (SIVmac239 g4,5), and SIVmneE11S. The vaccines were noninfectious, safe, and immunogenic, inducing antibody responses and cellular responses, including responses by CD8+ lymphocytes. Interpretation of protective efficacy following intrarectal challenge was complicated by incomplete take of the challenge in some SIV naive controls. C1 NCI, SAIC Frederick, AIDS Vaccine Program, Frederick, MD 21702 USA. Harvard Univ, New England Reg Primate Res Ctr, Sch Med, Southborough, MA 01772 USA. RP Lifson, JD (reprint author), NCI, SAIC Frederick, AIDS Vaccine Program, Bldg 535,5th Floor, Frederick, MD 21702 USA. RI Bess, Jr., Julian/B-5343-2012 FU PHS HHS [N01-C0-12400] NR 31 TC 26 Z9 27 U1 0 U2 0 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0047-2565 J9 J MED PRIMATOL JI J. Med. Primatol. PD AUG PY 2002 VL 31 IS 4-5 BP 205 EP 216 DI 10.1034/j.1600-0684.2002.02007.x PG 12 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 594AT UT WOS:000178027200008 PM 12390543 ER PT J AU Warren, J AF Warren, J TI Preclinical AIDS vaccine research: survey of SIV, SHIV, and HIV challenge studies in vaccinated nonhuman primates SO JOURNAL OF MEDICAL PRIMATOLOGY LA English DT Article; Proceedings Paper CT 19th Annual Symposium on Nonhuman Primate Models for AIDS CY NOV 07-10, 2001 CL SAN JUAN, PUERTO RICO SP BD Biosci Pharmingen, Glaxo, Res Ctr Minor Inst Program, Univ Puerto Rico, Med Sci Campus, Off Chancellor, Unit Comparat Med, Virol Lab, Dept Microl DE AIDS Vaccine Surveillance System (AVSS); chimpanzee(s); immunogen(s) intravenous; macaque(s); mucosal; protection ID SIMIAN IMMUNODEFICIENCY VIRUS; HIV-1/SIV CHIMERIC VIRUS; GENE-DELETED SHIVS; RHESUS MACAQUES; IMMUNE-RESPONSES; CYNOMOLGUS MONKEYS; MACACA-NEMESTRINA; DNA VACCINATION; NEUTRALIZING ANTIBODIES; PROTECTIVE IMMUNITY AB This current supplementary and systematic survey of 237 preclinical AIDS vaccine challenge/protection studies in nonhuman primates enumerates and broadly describes the recent status of different vaccine strategies in macaque and chimpanzee experimental models. Published studies since the previous survey were compiled and categorized by their vaccine types, challenge parameters, and challenge results. These models have supportively verified that some prophylactic vaccine approaches, though rarely preventing infection (which is observed in these models with some passively administered antibody-based vaccines), can control to some degree primate lentivirus replication and disease development, and this is encouraging because it places more potentially effective immunogens on the precipice for early clinical studies. Many of these promising approaches may benefit from more testing in mucosal challenge models, and resources will be needed to follow more of these partially protected vaccinees for longer periods. C1 NIAID, Vaccine & Prevent Res Program, Div Aids, Preclin Res & Dev Branch,NIH, Bethesda, MD 20892 USA. RP Warren, J (reprint author), NIAID, Vaccine & Prevent Res Program, Div Aids, Preclin Res & Dev Branch,NIH, 6700 B Rockledge Dr,Room 4104, Bethesda, MD 20892 USA. FU NIAID NIH HHS [N01-AI-45205] NR 85 TC 19 Z9 19 U1 0 U2 1 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0047-2565 J9 J MED PRIMATOL JI J. Med. Primatol. PD AUG PY 2002 VL 31 IS 4-5 BP 237 EP 256 DI 10.1034/j.1600-0684.2002.02010.x PG 20 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 594AT UT WOS:000178027200011 PM 12390546 ER PT J AU Muller, CE Thorand, M Qurishi, R Diekmann, M Jacobson, KA Padgett, WL Daly, JW AF Muller, CE Thorand, M Qurishi, R Diekmann, M Jacobson, KA Padgett, WL Daly, JW TI Imidazo[2,1-i]purin-5-ones and related tricyclic water-soluble purine derivatives: Potent A(2A)- and A(3)-adenosine receptor antagonists SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID ADENOSINE RECEPTOR; CAPILLARY-ELECTROPHORESIS; CONVENIENT SYNTHESIS; BETA-CYCLODEXTRIN; FUTURE DRUGS; ANALOGS; 8-STYRYLXANTHINES; LIGANDS AB A series of tricyclic imidazo[2,1-i]purinones and ring-enlarged analogues derived from xanthine derivatives have been prepared as adenosine receptor (AR) antagonists. In comparison with xanthines, the tricyclic compounds exhibit increased water solubility due to a basic nitrogen atom, which can be protonated under physiological conditions. Substituents were introduced that confer high affinity for A(2A) or A(3) ARs, respectively. A new capillary electrophoresis method was developed for the determination of the enantiomeric purity of selected chiral products using native and modified beta-cyclodextrins as chiral discriminators. The compounds were investigated in radioligand binding assays at rat brain A(1) and A(2A) ARs. Selected compounds were additionally investigated in radioligand binding assays at human recombinant A(3) ARs and in functional studies (adenylate cyclase assays) at A(1) ARs of rat fat cell membranes, A(2A) ARs of rat PC 12 cell membranes, and mouse A(2B) ARs of NIH 3T3 cell membranes. Structure activity relationships were similar to those of corresponding xanthine derivatives. The 2-styrylimidazopurinones were less potent at A2A ARs as compared to 8-styrylxanthine derivatives. The most potent compound at A2A ARs was (S)-1,4-dimethyl-8-ethyl-2-styrylimidazo[2,1-i]purinone (S-25) exhibiting a K-i value of 424 nM at rat A(2A) ARs. The compound was highly selective for A(2A) receptors vs A(1) and A(3) ARs. Selectivity vs A(2B) ARs, however, was low. Among the 1-unsubstituted 2-phenyl-imidazo[2,1-i]purin-5-one derivatives, very potent and highly selective antagonists for human A(3) ARs were identified. The most potent A3 antagonist of the present series was (R)-4-methyl-8-ethyl-2-phenyl-imidazo [2,1-i]purin-5-one (R-24) exhibiting a K-i value of 2.3 nM and high selectivity for A(3) receptors vs all other AR subtypes. C1 Univ Bonn, Pharmaceut Inst Poppelsdorf, D-5300 Bonn, Germany. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Muller, CE (reprint author), Univ Bonn, Pharmaceut Inst Poppelsdorf, D-5300 Bonn, Germany. RI Jacobson, Kenneth/A-1530-2009; Muller, Christa/C-7748-2014 OI Jacobson, Kenneth/0000-0001-8104-1493; Muller, Christa/0000-0002-0013-6624 NR 42 TC 59 Z9 62 U1 2 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD AUG 1 PY 2002 VL 45 IS 16 BP 3440 EP 3450 DI 10.1021/jm011093d PG 11 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 578VY UT WOS:000177142800012 PM 12139454 ER PT J AU Chong, HS Garmestani, K Ma, DS Milenic, DE Overstreet, T Brechbiel, MW AF Chong, HS Garmestani, K Ma, DS Milenic, DE Overstreet, T Brechbiel, MW TI Synthesis and biological evaluation of novel macrocyclic ligands with pendent donor groups as potential yttrium chelators for radioimmunotherapy with improved complex formation kinetics SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID IN-VIVO BIODISTRIBUTION; HUMAN TUMOR XENOGRAFT; MONOCLONAL-ANTIBODIES; CHX-DTPA; CONVENIENT SYNTHESIS; Y-90; ACID; STABILITY; SELECTION; ISOMERS AB Novel 1,4,7-triazacyclononane-N,N',N"-triacetic acid (NOTA) based octadentate ligands [2-(4,7-biscarboxymethyl[1,4,7]triazacyclononan-1-ylethyl)carbonylmethylamino] acetic acid tetrahydrochloride (1) and [3-(4,7-biscarboxymethyl[1,4,7]triazacyclononan-1-yl-propyl)carbonylmethylamino] acetic acid tetrahydrochloride (2) with pendent donor groups as potential yttrium. chelators for radioimmunotherapy (RIT) have been prepared via a convenient and high-yield cyclization route. The complexation kinetics of the novel chelates with Y(III) was investigated and compared to that of 1,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetic acid (DOTA), a macrocyclic chelating agent well recognized as forming very stable complexes with yttrium but also limited in usage because of slow Y(III) complex formation rates. The in vitro stability of the corresponding Y-88-labeled complexes in human serum was assessed by measuring the release of Y-88 from the complexes over 14 days. The in vivo biodistribution of Y-86-labeled 1 in mice was evaluated and compared to that of the Y-86-DOTA complex. Formation of the Y complex of 1 was significantly more rapid than that of either 2 or DOTA. Serum stability of the Y-88 complex formed with 1 was equivalent to the DOTA complex, while the complex formed with 2 proved to be significantly unstable. The results obtained from a biodistribution study indicate that the Y-86-1 complex possesses in vivo stability comparable to the analogous DOTA complex. C1 NCI, Chem Sect, Radiat Oncol Branch, NIH, Bethesda, MD 20892 USA. RP Brechbiel, MW (reprint author), NCI, Chem Sect, Radiat Oncol Branch, NIH, Bldg 10,Room B3B69, Bethesda, MD 20892 USA. NR 45 TC 44 Z9 44 U1 0 U2 7 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD AUG 1 PY 2002 VL 45 IS 16 BP 3458 EP 3464 DI 10.1021/jm0200759 PG 7 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 578VY UT WOS:000177142800014 PM 12139456 ER PT J AU Thomas, JB Atkinson, RN Namdev, N Rothman, RB Gigstad, KM Fix, SE Mascarella, SW Burgess, JP Vinson, NA Xu, H Dersch, CM Cantrell, BE Zimmerman, DM Carroll, FI AF Thomas, JB Atkinson, RN Namdev, N Rothman, RB Gigstad, KM Fix, SE Mascarella, SW Burgess, JP Vinson, NA Xu, H Dersch, CM Cantrell, BE Zimmerman, DM Carroll, FI TI Discovery of an opioid kappa receptor selective pure antagonist from a library of N-substituted 4 beta-methyl-5-(3-hydroxyphenyl)morphans SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID SUBTYPE-SELECTIVITY; (+)-(3R,4R)-DIMETHYL-4-(3-HYDROXYPHENYL)PIPERIDINE; IDENTIFICATION; AGONISTS; BINDING; POTENT; AGENTS AB A library of compounds biased toward opioid receptor antagonist activity was prepared by incorporating N-phenylpropyl-4p-methyl-5-(3-hydroxyphenyl)morphans as the core scaffold using simultaneous solution phase synthetic methodology. From this library, N-phenylpropyl-4beta-methyl-5-(3-hydroxyphenyl)-7alpha-[3-(1-piperidinyl)propanamido]morphan [(-)-3b] was identified as the first potent and selective k opioid receptor antagonist from the 5-phenylmorphan class of opioids. C1 Res Triangle Inst, Res Triangle Pk, NC 27709 USA. Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs, Indianapolis, IN USA. NIDA, Clin Psychopharmacol Sect, Addict Res Sect, Baltimore, MD 21224 USA. RP Carroll, FI (reprint author), Res Triangle Inst, POB 12194, Res Triangle Pk, NC 27709 USA. FU NIDA NIH HHS [DA 09045] NR 19 TC 21 Z9 23 U1 0 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD AUG 1 PY 2002 VL 45 IS 16 BP 3524 EP 3530 DI 10.1021/jm020084h PG 7 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 578VY UT WOS:000177142800021 PM 12139463 ER PT J AU Bagorda, A Guerra, L Di Sole, F Helmle-Kolb, C Favia, M Jacobson, KA Casavola, V Reshkin, SJ AF Bagorda, A Guerra, L Di Sole, F Helmle-Kolb, C Favia, M Jacobson, KA Casavola, V Reshkin, SJ TI Extracellular adenine nucleotides regulate Na+/H+ exchanger NHE3 activity in A6-NHE3 transfectants by a cAMP/PKA-dependent mechanism SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE P2Y receptors; A6 cells; Na+/H+ exchanger; ATP ID RENAL EPITHELIAL-CELLS; PROTEIN-KINASE-A; PURINERGIC RECEPTORS; INTRACELLULAR PH; APICAL MEMBRANE; PROXIMAL TUBULE; ION-TRANSPORT; H+ EXCHANGER; KIDNEY-CELLS; CL CURRENTS AB As potential autocrine or paracrine factors, extracellular nucleotides are known to be important regulators of renal ion transporters by activating cell surface receptors and intracellular signaling pathways. We investigated the influence of extracellular adenine nucleotides on Na+/H+ exchanger isoform 3 (NHE3) activity in A6-NHE3 cells. This is a polarized cell line obtained by stable transfection of A6 cells with the cDNA encoding the rat isoform of NHE3, which is expressed on the apical membrane. Basolateral addition of the P2Y(1) agonist, 2-Me-SADP, induced an inhibition of NHE3 activity, which was prevented by preincubation with selective P2Y(1) antagonists, MRS 2179 (N-6-methyl-2'-deoxyadenosine-3',5'-bisphosphate) and MRS 2286 (2-[2-(2-chloro-6-methylamino-purin-9-yl)-ethyl]-propane- 1,3-bisoxy(diammoniumphosphate)). NHE3 activity was also significantly inhibited by ATP and ATP-gamma-S but not by UTP. 2-Me-SADP induced a P2Y(1) antagonist-sensitive increase in both [Ca2+]i and cAMP production. Pre-incubation with a PKC inhibitor, Calphostin C, or the calcium chelator BAPTA-AM, had no effect on the 2-MeSADP-dependent inhibition of NHE3 activity, whereas this inhibition was reversed by either incubation with the PKA inhibitor H89 or by mutation of two PKA target serines (S552 and S605) on NHE3. Pre-incubation of the A6-NHE3 cells with the synthetic peptide, Ht31, which prevents the binding between AKAPs and the regulatory PKA subunits RII, also prevented the 2-MeSADP-induced inhibition of NHE3. We conclude that only the cAMP/PKA pathway is involved in the inhibition of NHE3 activity. C1 Univ Bari, Dept Gen & Environm Physiol, I-70126 Bari, Italy. Univ Gottingen, Dept Physiol & Pathophysiol, Div Vegetat Physiol & Pathophysiol, D-37073 Gottingen, Germany. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Casavola, V (reprint author), Univ Bari, Dept Gen & Environm Physiol, Via Amendola 165-A, I-70126 Bari, Italy. RI Jacobson, Kenneth/A-1530-2009; OI Jacobson, Kenneth/0000-0001-8104-1493; CASAVOLA, Valeria/0000-0003-4471-4948; Reshkin, Stephan/0000-0001-9757-5908 NR 51 TC 12 Z9 12 U1 1 U2 4 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0022-2631 J9 J MEMBRANE BIOL JI J. Membr. Biol. PD AUG 1 PY 2002 VL 188 IS 3 BP 249 EP 259 DI 10.1007/s00232-001-0189-8 PG 11 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 581HL UT WOS:000177285800007 PM 12181615 ER PT J AU Kohn, MC Melnick, RL AF Kohn, MC Melnick, RL TI Biochemical origins of the non-monotonic receptor-mediated dose-response SO JOURNAL OF MOLECULAR ENDOCRINOLOGY LA English DT Article ID ESTROGEN-RECEPTOR; ENVIRONMENTAL ESTROGENS; GENE-EXPRESSION; COACTIVATOR; LIGAND; YEAST; BINDING; ELEMENT; ALPHA; PUBERTY AB A mathematical model was created to examine how xenobiotic ligands that bind to nuclear receptor proteins may affect transcriptional activation of hormone-regulated genes. The model included binding of the natural ligand (e.g. hormone) and xenobiotic ligands to the receptor, binding of the liganded receptor to receptor-specific DNA response sequences, binding of co-activator or co-repressor proteins (Rp) to the resulting complex, and the consequent transcriptional rate relative to that in the absence of the xenobiotic agent. The model predicted that the xenobiotic could act as a pure agonist, a pure antagonist, or a mixed agonist whose dose-response curve exhibits a local maximum. The response to the agent depends on the affinity of the liganded receptor-DNA complex for binding additional transcription factors (e.g. co-activator proteins). An inverted U-shaped dose-response occurred when basal levels of the natural ligand did not saturate receptor binding sites and the affinity for co-activator is weaker when the xenobiotic ligand is bound to the receptor than when the endogenous ligand is bound. The dose-response curve shape was not dependent on the affinity of the receptor for the xenobiotic agent; alteration of this value merely shifted the curve along the concentration axis. The amount of receptor, the density of DNA response sequences, and the affinity of the DNA-bound receptor for Rp determine the amplitude of the computed response with little overall change in curve shape. This model indicates that a non-monotonic dose-response is a plausible outcome for xenobiotic agents that activate nuclear receptors in the same manner as natural ligands. C1 Natl Inst Environm Hlth Sci, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. RP Kohn, MC (reprint author), Natl Inst Environm Hlth Sci, Environm Toxicol Program, POB 12233,MD A3-06, Res Triangle Pk, NC 27709 USA. NR 26 TC 35 Z9 36 U1 1 U2 4 PU SOC ENDOCRINOLOGY PI BRISTOL PA 17/18 THE COURTYARD, WOODLANDS, BRADLEY STOKE, BRISTOL BS32 4NQ, ENGLAND SN 0952-5041 J9 J MOL ENDOCRINOL JI J. Mol. Endocrinol. PD AUG PY 2002 VL 29 IS 1 AR UNSP 0952-5041/02/029-113 DI 10.1677/jme.0.0290113 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 589LY UT WOS:000177760900010 PM 12200233 ER PT J AU Perelygin, AA Kondrashov, FA Rogozin, IB Brinton, MA AF Perelygin, AA Kondrashov, FA Rogozin, IB Brinton, MA TI Evolution of the mouse polyubiquitin-C gene SO JOURNAL OF MOLECULAR EVOLUTION LA English DT Article DE concerted evolution; purifying selection; ubiquitin; multigene family; unequal crossing-over; apparent selection ID CONCERTED EVOLUTION; PROTEIN-DEGRADATION; MULTIGENE FAMILIES; UNEQUAL CROSSOVER; UBIQUITIN GENES; MUTATION-RATES; DNA-SEQUENCES; LOCUS; DROSOPHILA; SELECTION AB The polymeric ubiquitin (poly-u) genes are composed of tandem 228-bp repeats with no spacer sequences between individual monomer units. Ubiquitin is one of the most conserved proteins known to date, and the individual units within a number of poly-u genes are significantly more similar to each other than would be expected if each unit evolved independently. It has been proposed that the rather striking similarity among poly-u monomers in some lineages is caused by a series of homogenization events. Here we report the sequences of the polyubiquitin-C (Ubc) genes in two mouse strains. Analysis of these sequences, as well as those of the previously reported Chinese hamster and rat poly-u genes, supports the assertion that the homogenization of the ubiquitin-C gene in rodents is due to unequal crossing-over events. The sequence divergence of noncoding DNA was used to estimate the frequency of unequal crossing-over events (6.3 x 10(-5) events per generation) in the Ubc gene, as well as to provide evidence of apparent selection in the poly-u gene. C1 Georgia State Univ, Dept Biol, Atlanta, GA 30303 USA. NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20892 USA. RP Perelygin, AA (reprint author), Georgia State Univ, Dept Biol, 24 Peachtree Ctr Ave, Atlanta, GA 30303 USA. RI Kondrashov, Fyodor Alexeevich/H-6331-2015 OI Kondrashov, Fyodor Alexeevich/0000-0001-8243-4694 FU NIAID NIH HHS [AI 45135] NR 41 TC 8 Z9 9 U1 1 U2 3 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0022-2844 J9 J MOL EVOL JI J. Mol. Evol. PD AUG PY 2002 VL 55 IS 2 BP 202 EP 210 DI 10.1007/s00239-002-2318-0 PG 9 WC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity GA 570ZY UT WOS:000176691400007 PM 12107596 ER PT J AU Gozes, I Alcalay, R Giladi, E Pinhasov, A Furman, S Brenneman, DE AF Gozes, I Alcalay, R Giladi, E Pinhasov, A Furman, S Brenneman, DE TI NAP accelerates the performance of normal rats in the water maze SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE peptides; neuroprotection; middle-aged rats ID DEPENDENT NEUROPROTECTIVE PROTEIN; VASOACTIVE-INTESTINAL-PEPTIDE; NEURONAL CULTURES; VIP; CELLS AB NAP (Asn-Ala-Pro-Val-Ser-Ile-Pro-Gln) has neuroprotective, memory enhancing, and neurotrophic properties. NAP is a short peptide sequence derived from the recently cloned, activity-dependent neuroprotective protein. The current study was designed to evaluate NAP activity in normal middle-aged animals to further assess NAP's breadth of neuroprotection. NAP was administered by inhalation. Results showed that in the paradigm of the Morris water maze, assessing short-term memory, only the NAP-treated middle-aged rats and not placebo-treated rats showed significant improvements by the end of the testing period. These results suggest efficacy for NAP in normal aging that is associated with accumulating environmental and genetic toxic factors. C1 Tel Aviv Univ, Sackler Fac Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NICHD, Sec Dev & Mol Pharmacol, LDN, NIH, Bethesda, MD 20892 USA. RP Gozes, I (reprint author), Tel Aviv Univ, Sackler Fac Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NR 17 TC 29 Z9 29 U1 0 U2 1 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0895-8696 J9 J MOL NEUROSCI JI J. Mol. Neurosci. PD AUG-OCT PY 2002 VL 19 IS 1-2 BP 167 EP 170 DI 10.1007/s12031-002-0028-0 PG 4 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 590AJ UT WOS:000177794700027 PM 12212775 ER PT J AU Akbar, M Kim, HY AF Akbar, M Kim, HY TI Protective effects of docosahexaenoic acid in staurosporine-induced apoptosis: involvement of phosphatidylinositol-3 kinase pathway SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE Akt; apoptosis; docosahexaenoic acid; neuronal cells; phosphatidylserine ID POLYUNSATURATED FATTY-ACIDS; PROTEIN-KINASE; PEROXISOMAL DISORDERS; EICOSAPENTAENOIC ACID; NEURONAL APOPTOSIS; CELLS; PHOSPHORYLATION; DEATH; BRAIN; AKT AB Docosahexaenoic acid (22:6n-3, DHA) is highly enriched in neuronal membranes and is considered to be essential for proper brain function. We have previously demonstrated in Neuro 2A cells that DHA as a membrane component protects cells from apoptotic death induced by serum deprivation (Kim et al. 2000). In the present study we demonstrate that staurosporine (ST) induces apoptosis in Neuro 2A cells and DHA enrichment prior to the ST treatment significantly inhibits the apoptotic cell death, as evidenced by the reduction of caspase-3 activity, cleavage of pro-caspase-3 to active caspase-3, DNA strand-breaking and laddering. Enrichment of cells with other fatty acids such as oleic and arachidonic acids did not exert such an effect, indicating that the antiapoptotic effect was specific to DHA enrichment. Among the several protein kinase inhibitors, only phosphatidylinositol 3-kinase (PI3-K) inhibitors, wortmanin, and LY-294002 abolished the protective effect of DHA in ST-induced apoptosis. Concurrently, ST-treatment significantly decreased the phosphorylation status of Akt at Ser-473 and Thr-308 as well as Akt activity, and this reduction was partially prevented by DHA enrichment. The extent of the antiapoptotic effect of DHA correlated with a time-dependent increase in the phosphatidylserine (PS) content upon DHA enrichment. When cells were enriched with DHA in serine-free medium, the PS increase diminished and the DHA effect on caspase-3 activation as well as Akt phosphorylation in ST-induced apoptosis was no longer apparent, suggesting that DHA's role in accumulating membrane PS is an important component for the observed protection. In summary, DHA enrichment uniquely protects ST-induced apoptosis in a PS- and PI3-K-dependent manner. From these data, we suggest that the antiapoptotic effect of DHA is mediated at least in part through the PI3-K/Akt pathway, facilitated by DHA-induced PS accumulation. C1 NIAAA, Sect Mass Spectrometry, LMBB, NIH, Rockville, MD 20852 USA. RP Kim, HY (reprint author), NIAAA, Sect Mass Spectrometry, LMBB, NIH, 12420 Parklawn Dr, Rockville, MD 20852 USA. NR 46 TC 85 Z9 90 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD AUG PY 2002 VL 82 IS 3 BP 655 EP 665 DI 10.1046/j.1471-4159.2002.01015.x PG 11 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 576VT UT WOS:000177027400020 PM 12153489 ER PT J AU Conant, K Haughey, N Nath, A St Hillaire, C Gary, DS Pardo, CA Wahl, LM Bilak, M Milward, E Mattson, MP AF Conant, K Haughey, N Nath, A St Hillaire, C Gary, DS Pardo, CA Wahl, LM Bilak, M Milward, E Mattson, MP TI Matrix metalloproteinase-1 activates a pertussis toxin-sensitive signaling pathway that stimulates the release of matrix metalloproteinase-9 SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE CD47; G protein-coupled receptor; integrins; MMP-1; MMP-9 ID RECEPTOR-RELATED PROTEIN; IN-VITRO; INTEGRIN; CD47; EXPRESSION; CELLS; BETA; TAT; ANGIOGENESIS; INVOLVEMENT AB The matrix metalloproteinases (MMPs) are a family of structurally related metalloendopeptidases so named due to their propensity to target extracellular matrix (ECM) proteins. Accumulating evidence, however, suggests that these proteases cleave numerous non-ECM substrates including enzymes and cell surface receptors. MMPs may also bind to cell surface receptors, though such binding has typically been thought to mediate internalization and degradation of the bound protease. More recently, it has been shown that MMP-1 coimmunoprecipitates with the alpha(2) beta(1) integrin, a receptor for collagen. This association may serve to localize the enzymatic activity of MMP-1 so that collagen is cleaved and cell migration is facilitated. In other studies, however, it has been shown that integrin engagement may be linked to the activation of signaling cascades including those mediated by Gialpha containing heterotrimers. As an example, alpha(2) beta(1) can form a complex with CD47 that may associate with Gialpha. In the present study we have therefore investigated the possibility that MMP-1 may affect intracellular changes that are linked to the activation of a Gi protein-coupled receptor. We show that treatment of neural cells with MMP-1 is followed by a rapid reduction in cytosolic levels of cAMP. Moreover, MMP-1 potentiates proteinase activated receptor-1 (PAR-1) agonist-linked increases in intracellular calcium, an effect which is often observed when an agonist of a Gi protein-coupled receptor is administered in association with an agonist of a Gq coupled receptor. In addition, MMP-1 stimulates pertussis toxin sensitive release ofMMP-9 both from cultured neural cells and monocyte/macrophages. Together, these results suggest that MMP-1 signals through a pertussis toxin-sensitive G protein-coupled receptor. C1 Johns Hopkins Univ, Sch Med, Dept Neurol, Baltimore, MD 21287 USA. NIA, Neurosci Lab, Baltimore, MD 21224 USA. Univ Kentucky, Dept Neurol, Lexington, KY 40536 USA. Natl Inst Dent & Craniofacial Res, Immunopathol Sect, Bethesda, MD USA. RP Conant, K (reprint author), Johns Hopkins Univ, Sch Med, Dept Neurol, Meyer 6-181,600 N Wolfe St, Baltimore, MD 21287 USA. RI Mattson, Mark/F-6038-2012 FU NIMH NIH HHS [MH 63722] NR 48 TC 22 Z9 23 U1 1 U2 3 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD AUG PY 2002 VL 82 IS 4 BP 885 EP 893 DI 10.1046/j.1471-4159.2002.01038.x PG 9 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 582UJ UT WOS:000177369300017 PM 12358794 ER PT J AU Wang, Y Mattson, MP Furukawa, K AF Wang, Y Mattson, MP Furukawa, K TI Endoplasmic reticulum calcium release is modulated by actin polymerization SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE actin; calcium; endoplasmic reticulum; inositol trisphosphate; neuron; ryanodine ID LONG-TERM POTENTIATION; HIPPOCAMPAL-NEURONS; INTRACELLULAR CA2+; NEURITE OUTGROWTH; CELL-CULTURE; F-ACTIN; CHANNEL; DEPOLYMERIZATION; PLASTICITY; EXCITOTOXICITY AB Intracellular calcium ions regulate the structure and functions of cytoskeletal proteins. On the other hand, recent studies have shown that the cytoskeleton, and actin filaments in particular, can modulate calcium influx through plasma membrane ligand- and voltage-gated channels. We now report that calcium release from inositol trisphosphate (IP3) and ryanodine-sensitive endoplasmic reticulum (ER) stores is modulated by polymerization and depolymerization of actin filaments in cultured hippocampal neurons. Depolymerization of actin filaments with cytochalasin D attenuates calcium release induced by carbamylcholine (CCh; a muscarinic agonist for IP3 pathway), caffeine (a ryanodine receptor agonist) and thapsigargin (an inhibitor of the ER calcium- ATPase) in both the presence and absence of extracellular calcium. Conversely, the actin polymerizing agent jasplakinolide potentiates calcium release induced by CCh, caffeine and thapsigargin. Cytochalasin D attenuated, while jasplakinolide augmented, thapsigargin-induced JNK activation and neuronal cell death. Our data show that the actin cytoskeleton regulates ER calcium release, suggesting roles for actin in the various physiological and pathological processes that involve calcium release. C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Furukawa, K (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 35 TC 50 Z9 50 U1 0 U2 2 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD AUG PY 2002 VL 82 IS 4 BP 945 EP 952 DI 10.1046/j.1471-4159.2002.01059.x PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 582UJ UT WOS:000177369300023 PM 12358800 ER PT J AU Pedersen, WA Chan, SL Zhu, HY Abdur-Rahman, LA Verdi, JM Mattson, MP AF Pedersen, WA Chan, SL Zhu, HY Abdur-Rahman, LA Verdi, JM Mattson, MP TI Numb isoforms containing a short PTB domain promote neurotrophic factor-induced differentiation and neurotrophic factor withdrawal-induced death of PC12 cells SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE apoptosis; MAP kinase; neurite outgrowth; phosphotyrosine-binding domain; proline-rich region; TrkA ID NERVE GROWTH-FACTOR; MOUSE CORTICAL NEUROGENESIS; PROGRAMMED NEURONAL DEATH; CYTOCHROME-C RELEASE; MAMMALIAN NUMB; PHEOCHROMOCYTOMA CELLS; ASYMMETRIC LOCALIZATION; HIPPOCAMPAL-NEURONS; CASPASE INHIBITION; NEURITE OUTGROWTH AB The development of the nervous system is regulated by trophic signals that control cell proliferation, differentiation, and survival. Numb is an evolutionarily conserved protein identified by its ability to control cell fate in the nervous system of Drosophila . Mammals express four isoforms of Numb that differ in the length of a phosphotyrosine-binding (PTB) domain and a proline-rich region (PRR). Using PC12 cells stably expressing each of the human isoforms, we show that Numb regulates sensitivity of the cells to neurotrophic factor-induced differentiation and neurotrophic factor withdrawal-induced death in an isoform-specific manner. Numb isoforms containing a short PTB domain enhance the differentiation response to NGF and enhance apoptosis upon NGF withdrawal; Numb isoforms containing a long PTB domain exhibit the same sensitivity to NGF as vector-transfected cells. These effects of Numb were found to be independent of the length of the PRR. In undifferentiated conditions, the levels of full-length TrkA and of phosphorylated p44/p42 mitogen-activated protein kinase (MAPK) are increased in cells expressing Numb isoforms with a short PTB domain, indicating an up-regulation of NGF signaling pathways. Furthermore, we provide evidence that the mechanism whereby short PTB domain Numb isoforms sensitize cells to trophic factor deprivation-induced apoptosis involves elevations in intracellular calcium concentrations. Our results suggest that Numb sensitizes cells to neurotrophin responses in an isoform-specific manner, an effect that may play an important role in the development and plasticity of the nervous system. C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. Johns Hopkins Bayview Med Ctr, Baltimore, MD USA. Univ Kentucky, Sanders Brown Ctr Aging, Lexington, KY 40536 USA. Robarts Res Inst, London, ON N6A 5C1, Canada. Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr 4F02, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 59 TC 17 Z9 18 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD AUG PY 2002 VL 82 IS 4 BP 976 EP 986 DI 10.1046/j.1471-4159.2002.01036.x PG 11 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 582UJ UT WOS:000177369300026 PM 12358803 ER PT J AU Agarwal, RK Sun, SH Su, SB Chan, CC Caspi, RR AF Agarwal, RK Sun, SH Su, SB Chan, CC Caspi, RR TI Pertussis toxin alters the innate and the adaptive immune responses in a pertussis-dependent model of autoimmunity SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE autoimmune uveitis; EAU; chemokines; pertussis toxin ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; RETINOID-BINDING PROTEIN; B10.A MICE; T-CELLS; UVEORETINITIS; DISEASE; INDUCTION; PEPTIDE; IRBP; SUSCEPTIBILITY AB Pertussis toxin (PTX) is used to promote development of autoimmune diseases. The mechanism(s) are still incompletely understood. We dissected the innate and adaptive immune responses in a PTX-dependent model of autoimmune retinal disease, experimental autoimmune uveoretinitis (EAU), a Th1-driven disease of the neural retina elicited in F344 rats with a peptide derived from the retinal antigen interphotoreceptor retinoid binding protein (IRBP). Our results showed that optimal doses of PTX led to strongly increased innate cytokine responses, followed by enhanced adaptive Th1 immunity and disease. At supraoptimal doses of PTX, EAU was suppressed, the animals exhibited persistent lymphocytosis and had an inhibited chemotactic response to chemokines. We suggest that the suppressive effect of PTX at supraoptimal doses is due to inhibition of lymphocyte emigration from the blood into the target tissue, secondary to inhibition of Gi-protein-coupled chemokine receptor signaling, that persists into the effector phase of disease. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Caspi, RR (reprint author), NEI, Immunol Lab, NIH, 10 Ctr Dr,10-10N222, Bethesda, MD 20892 USA. EM rcaspi@helix.nih.gov NR 38 TC 21 Z9 24 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-5728 J9 J NEUROIMMUNOL JI J. Neuroimmunol. PD AUG PY 2002 VL 129 IS 1-2 BP 133 EP 140 AR PII S0165-5728(02)00203-5 DI 10.1016/S0165-5728(02)00203-5 PG 8 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 590BQ UT WOS:000177797700016 PM 12161029 ER PT J AU Sakurai, K Zou, JP Tschetter, JR Ward, JM Shearer, GM AF Sakurai, K Zou, JP Tschetter, JR Ward, JM Shearer, GM TI Effect of indoleamine 2,3-dioxygenase on induction of experimental autoimmune encephalomyelitis SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE experimental autoimmune encephalomyelitis; cytokines; indoleamine 2,3-dioxygenase; myelin basic protein; Th1; Th2 ID CENTRAL-NERVOUS-SYSTEM; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; MYELIN OLIGODENDROCYTE GLYCOPROTEIN; MESSENGER-RNA EXPRESSION; T-CELL PROLIFERATION; MULTIPLE-SCLEROSIS; TRYPTOPHAN CATABOLISM; INTERFERON-GAMMA; IFN-GAMMA; GENE-EXPRESSION AB Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated demyelinating disease of the central nervous system (CNS). Indoleamine 2,3-dioxygenase (IDO) is an enzyme that catabolizes tryptophan, which can result in the death of T lymphocytes. This effect of IDO is inhibited by 1-methyl-tryptophan (1-MT). We used a murine model of EAE to demonstrate; (1) opposing patterns of spinal cord IDO and interferon-gamma (INF-gamma) mRNA expression through the preclinical, acute and remission I phases of EAE; (2) a change in the kynurenine-to-tryptophan (KIT) ratio during these same phases; and (3) 1-MT-induced exacerbation of clinical and histologic disease parameters during EAE. These results suggest that IDO may contribute to the regulation of T cell activity associated with the different phases of this animal model of multiple sclerosis (MS). (C) 2002 Elsevier Science B.V. All rights reserved. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NCI, NIH, Frederick, MD 21702 USA. RP Shearer, GM (reprint author), NCI, Expt Immunol Branch, NIH, Bldg 10,Room 4B36,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Tschetter, Jolynne/0000-0003-1743-1221 NR 39 TC 138 Z9 139 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-5728 J9 J NEUROIMMUNOL JI J. Neuroimmunol. PD AUG PY 2002 VL 129 IS 1-2 BP 186 EP 196 AR PII S0165-5728(02)00176-5 DI 10.1016/S0165-5728(02)00176-5 PG 11 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 590BQ UT WOS:000177797700022 PM 12161035 ER PT J AU Leigh, RJ Ramat, S Zee, DS AF Leigh, RJ Ramat, S Zee, DS TI High frequency saccadic osscillations are generated by brain stem, not cerebellar, neurons SO JOURNAL OF NEUROLOGY NEUROSURGERY AND PSYCHIATRY LA English DT Meeting Abstract C1 Case Western Reserve Univ, Cleveland, OH 44106 USA. Johns Hopkins Univ, Baltimore, MD 21218 USA. NEI, Bethesda, MD 20892 USA. RI Ramat, Stefano/E-6495-2011 OI Ramat, Stefano/0000-0001-5932-186X NR 0 TC 0 Z9 0 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0022-3050 J9 J NEUROL NEUROSUR PS JI J. Neurol. Neurosurg. Psychiatry PD AUG PY 2002 VL 73 IS 2 BP 220 EP 220 PG 1 WC Clinical Neurology; Psychiatry; Surgery SC Neurosciences & Neurology; Psychiatry; Surgery GA 577KK UT WOS:000177059900067 ER PT J AU Solinas, M Ferre, S You, ZB Karcz-Kubicha, M Popoli, P Goldberg, SR AF Solinas, M Ferre, S You, ZB Karcz-Kubicha, M Popoli, P Goldberg, SR TI Caffeine induces dopamine and glutamate release in the shell of the nucleus accumbens SO JOURNAL OF NEUROSCIENCE LA English DT Article DE caffeine; adenosine; dopamine; glutamate; accumbens; microdialysis ID EXTRACELLULAR DOPAMINE; ADENOSINE RECEPTORS; LOCOMOTOR-ACTIVITY; BASAL GANGLIA; D-AMPHETAMINE; RAT STRIATUM; CORE; SYSTEM; MICRODIALYSIS; ANTAGONISTS AB An increase in the extracellular concentration of dopamine in the nucleus accumbens (NAc) is believed to be one of the main mechanisms involved in the rewarding and motor-activating properties of psychostimulants such as amphetamines and cocaine. Using in vivo microdialysis in freely moving rats, we demonstrate that systemic administration of behaviorally relevant doses of caffeine can preferentially increase extracellular levels of dopamine and glutamate in the shell of the NAc. These effects could be reproduced by the administration of a selective adenosine A1 receptor antagonist but not by a selective adenosine A2A receptor antagonist. This suggests that caffeine, because of its ability to block adenosine A1 receptors, shares neurochemical properties with other psychostimulants, which could contribute to the widespread consumption of caffeine-containing beverages. C1 NIDA, NIH, Intramural Res Program, Behav Neurosci Branch,Preclin Pharmacol Sect, Baltimore, MD 21224 USA. NIDA, NIH, Intramural Res Program, Behav Neurosci Branch,Sect Behav Neurosci, Baltimore, MD 21224 USA. Ist Super Sanita, Dept Pharmacol, I-00161 Rome, Italy. RP Goldberg, SR (reprint author), NIDA, NIH, Intramural Res Program, Behav Neurosci Branch,Preclin Pharmacol Sect, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Popoli, Patrizia/B-5397-2008; Ferre, Sergi/K-6115-2014; Solinas, Marcello/M-3500-2016 OI Ferre, Sergi/0000-0002-1747-1779; Solinas, Marcello/0000-0002-0664-5964 NR 25 TC 121 Z9 122 U1 1 U2 16 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD AUG 1 PY 2002 VL 22 IS 15 BP 6321 EP 6324 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 579MV UT WOS:000177182700004 PM 12151508 ER PT J AU Zhang, WL Yamada, M Gomeza, J Basile, AS Wess, J AF Zhang, WL Yamada, M Gomeza, J Basile, AS Wess, J TI Multiple muscarinic acetylcholine receptor subtypes modulate striatal dopamine release, as studied with M-1-M-5 muscarinic receptor knock-out mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE acetylcholine; dopamine release; knock-out mice; muscarinic receptors; oxotremorine; striatum ID IN-VIVO MICRODIALYSIS; CAT CAUDATE-NUCLEUS; BASAL GANGLIA; RAT-BRAIN; NEUROTRANSMITTER RELEASE; MEDIATED MODULATION; M2-MUSCARINIC DRUGS; NERVE-TERMINALS; MESSENGER-RNAS; GABA RELEASE AB A proper balance between striatal muscarinic cholinergic and dopaminergic neurotransmission is required for coordinated locomotor control. Activation of striatal muscarinic acetylcholine receptors (mAChRs) is known to modulate striatal dopamine release. To identify the mAChR subtype(s) involved in this activity, we used genetically altered mice that lacked functional M-1-M-5 mAChRs [knock-out (KO) mice]. In superfused striatal slices from wild-type mice, the non-subtype-selective muscarinic agonist oxotremorine led to concentration-dependent increases in potassium-stimulated [H-3] dopamine release (by up to 60%). The lack of M-1 or M-2 receptors had no significant effect on the magnitude of these responses. Strikingly, oxotremorine-mediated potentiation of stimulated striatal [H-3] dopamine release was abolished in M-4 receptor KO mice, significantly increased in M-3 receptor-deficient mice, and significantly reduced (but not abolished) in M-5 receptor KO mice. Additional release studies performed in the presence of tetrodotoxin suggested that the dopamine release-stimulating M-4 receptors are probably located on neuronal cell bodies, but that the release-facilitating M-5 and the release-inhibiting M-3 receptors are likely to be located on nerve terminals. Studies with the GABA(A) receptor blocker bicuculline methochloride suggested that M-3 and M-4 receptors mediate their dopamine release-modulatory effects via facilitation or inhibition, respectively, of striatal GABA release. These results provide unambiguous evidence that multiple mAChR subtypes are involved in the regulation of striatal dopamine release. These findings should contribute to a better understanding of the important functional roles that the muscarinic cholinergic system plays in striatal function. C1 NIDDKD, Mol Signalling Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP NIDDKD, Mol Signalling Sect, Bioorgan Chem Lab, NIH, Bldg 8A,Room B1A-05,8 Ctr Dr MSC 0810, Bethesda, MD 20892 USA. EM jwess@helix.nih.gov NR 42 TC 100 Z9 103 U1 1 U2 2 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD AUG 1 PY 2002 VL 22 IS 15 BP 6347 EP 6352 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 579MV UT WOS:000177182700008 PM 12151512 ER PT J AU Morales, M Wng, SD AF Morales, M Wng, SD TI Differential composition of 5-hydroxytryptamine(3) receptors synthesized in the rat CNS and peripheral nervous system SO JOURNAL OF NEUROSCIENCE LA English DT Article DE 5-HT3A subunit; 5-HT3B subunit; myenteric plexus; nodose ganglia; superior cervical ganglia; serotonin receptors ID NUCLEUS TRACTUS SOLITARIUS; SUPERIOR CERVICAL-GANGLION; 5-HT3 RECOGNITION SITES; FUNCTIONAL EXPRESSION; SEROTONIN RECEPTOR; GABAERGIC NEURONS; INDUCED EMESIS; BINDING-SITES; H-3 ZACOPRIDE; VAGUS NERVE AB The type 3 serotonin (5-HT3) receptor is the only ligand-gated ion channel receptor for serotonin in vertebrates. Two 5-HT3 receptor subunits have been cloned, subunit A (5-HT3A) and subunit B (5-HT3B). We used in situ hybridization histochemistry and reverse transcriptase-PCR amplification to demonstrate that 5-HT3A subunit transcripts are expressed in central and peripheral neurons. In contrast, 5-HT3B subunit transcripts are restricted to peripheral neurons. Thus, the prevalent form of 5-HT3 receptor synthesized within the CNS lacks the 5-HT3B subunit. Because coexpression of 5-HT3A and 5-HT3B subunits produces heteromeric 5-HT3A/3B receptors with properties that differ from those of 5-HT3A homomeric receptors, we investigated possible coexpression of both subunits at the cellular level. We found that near to 90% of all 5-HT3B expressing neurons coexpress the 5-HT3A subunit in superior cervical and nodose ganglia (NG). In addition, there is a cellular population that expresses only the 5-HT3A subunit. Therefore, peripheral neurons have the capacity to synthesize two different 5-HT3 receptors, 5-HT3A+(/3B)- and 5-HT3A+/(3B)- receptors. We also determined that neurons of NG projecting to the nucleus tractus solitarium and those of dorsal root ganglia projecting to superficial layers of the spinal cord express 5-HT3A or 5-HT3A/3B subunits. Thus, presynaptic 5-HT3 receptors containing the 5-HT3B subunit might be present in these target brain areas. The compartmentalized structural composition of the 5-HT3 receptor may be the basis of functional diversity within this receptor. This raises the possibility that 5-HT3 receptors participating in sympathetic, parasympathetic and sensory functions may be functionally different from those involved in cognition and emotional behavior. C1 NIDA, Baltimore, MD 21224 USA. Natl Def Med Ctr, Dept Biol & Anat, Taipei 114, Taiwan. RP Morales, M (reprint author), NIDA, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. EM mmorales@intra.nida.nih.gov NR 49 TC 121 Z9 125 U1 0 U2 2 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD AUG 1 PY 2002 VL 22 IS 15 BP 6732 EP 6741 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 579MV UT WOS:000177182700048 PM 12151552 ER PT J AU Pace, JR Lonser, RR Kirkby, RD Jeffries, N Rogawski, MA Oldfield, EH AF Pace, JR Lonser, RR Kirkby, RD Jeffries, N Rogawski, MA Oldfield, EH TI Epileptiform activity extinguished by amygdala infusion of the neurotoxin ibotenate in a rat model of temporal lobe epilepsy SO JOURNAL OF NEUROSURGERY LA English DT Article DE ibotenate; epilepsy; convection; amygdala; rat ID KAINIC ACID; BRAIN; DELIVERY; SEIZURES; NEURONS; LESIONS AB Object. The long-term antiseizure effects of local convection-enhanced infusion of the excitotoxin ibotenate were examined in a rat model of temporal lobe epilepsy. Methods. A single injection of kainate, an epileptogenic excitatory amino acid, into the left amygdala elicited chronic spontaneous recurrent seizure activity for at least 36 days after the injection. Two weeks after the injection, infusion of ibotenate, a nonepileptogenic excitatory amino acid that is an axon-sparing neuronal cell toxin, into the left amygdala and piriform lobe induced immediate and permanent extinction of electrical and behavioral seizure activity. Conclusions. Lesioning of an epileptic focus by convective distribution of ibotenate can produce an enduring suppression of seizure activity, indicating a chemical neurosurgical approach for epilepsy therapy. C1 NINDS, Surg Neurol Branch, Epilepsy Res Sect, NIH, Bethesda, MD 20892 USA. NINDS, Biostat Branch, NIH, Bethesda, MD 20892 USA. RP Oldfield, EH (reprint author), NINDS, Surg Neurol Branch, Epilepsy Res Sect, NIH, Room 5D37,Bldg 10, Bethesda, MD 20892 USA. RI Kipke, Daryl/A-2167-2009 NR 20 TC 3 Z9 4 U1 0 U2 0 PU AMER ASSOC NEUROLOGICAL SURGEONS PI CHARLOTTESVILLE PA UNIV VIRGINIA, 1224 WEST MAIN ST, STE 450, CHARLOTTESVILLE, VA 22903 USA SN 0022-3085 J9 J NEUROSURG JI J. Neurosurg. PD AUG PY 2002 VL 97 IS 2 BP 450 EP 454 DI 10.3171/jns.2002.97.2.0450 PG 5 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 582LP UT WOS:000177352700028 PM 12186475 ER PT J AU Salinas-Torres, VH Pereira, CAB Tiwari, RC AF Salinas-Torres, VH Pereira, CAB Tiwari, RC TI Bayesian nonparametric estimation in a series system or a competing-risks model SO JOURNAL OF NONPARAMETRIC STATISTICS LA English DT Article DE Dirichlet processes; Dirichlet multivariate processes; randomly censored data; cumulative hazard rate; product-integration ID SURVIVAL CURVES; LARGE SAMPLE AB This article presents a Bayesian nonparametric approach to the estimation of a system and its components' survival functions arising from observing the failure of a series system or a competing risk model. A Dirichlet multivariate process is used as a prior for the vector of the components' random subsurvival function to derive Bayes estimator of the survival function when the cause of failure belongs to a certain risk subset. This is done as follows. First, Peterson's formula is evaluated using the Bayes estimators of the subsurvival functions corresponding to the risk subset, to obtain a plugged-in nonparametric estimator of the survival function associated with the risk subset. Then, using the product-integration approach, it is proved that this nonparametric estimator is in fact the Bayes estimator of the survival function corresponding to the risk subset under quadratic loss function and the Dirichlet multivariate process. The weak convergence and the strong consistency of the estimator is established. The special case when the system has only two components corresponds to well studied randomly censored model. C1 Univ Santiago, Dept Math, Santiago, Chile. Univ Sao Paulo, Dept Stat, BR-05315970 Sao Paulo, SP, Brazil. Univ N Carolina, Dept Math, Charlotte, NC 28223 USA. RP Tiwari, RC (reprint author), NCI, SRAB, DCCPS, 6116 Execut Blvd,Room 5039, Bethesda, MD 20892 USA. NR 14 TC 19 Z9 19 U1 1 U2 1 PU TAYLOR & FRANCIS LTD PI ABINGDON PA 4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLAND SN 1048-5252 J9 J NONPARAMETR STAT JI J. Nonparametr. Stat. PD AUG PY 2002 VL 14 IS 4 BP 449 EP 458 DI 10.1080/10485250290024503 PG 10 WC Statistics & Probability SC Mathematics GA 586CT UT WOS:000177566900008 ER PT J AU Price, JC Drevets, WC Ruszkiewicz, J Greer, PJ Villemagne, VL Xu, L Mazumdar, S Cantwell, MN Mathis, CA AF Price, JC Drevets, WC Ruszkiewicz, J Greer, PJ Villemagne, VL Xu, L Mazumdar, S Cantwell, MN Mathis, CA TI Sequential (H2OPET)-O-15 studies in baboons: Before and after amphetamine SO JOURNAL OF NUCLEAR MEDICINE LA English DT Article DE PET; blood flow; amphetamine ID CEREBRAL BLOOD-FLOW; POSITRON-EMISSION-TOMOGRAPHY; INDUCED DOPAMINE RELEASE; C-11 RACLOPRIDE BINDING; IN-VIVO; BRAIN; PET; PERFUSION; INVIVO; SCHIZOPHRENIA C1 Univ Pittsburgh, Sch Med, Dept Radiol, PET Facil, Pittsburgh, PA 15213 USA. NIMH, Mood & Anxiety Disorders Program, Bethesda, MD 20892 USA. Univ Pittsburgh, Sch Med, Dept Biostat, Pittsburgh, PA USA. RP Price, JC (reprint author), Univ Pittsburgh, Sch Med, Dept Radiol, PET Facil, 200 Lothrop ST,Room B938, Pittsburgh, PA 15213 USA. RI Mathis, Chester/A-8607-2009 NR 38 TC 20 Z9 20 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD AUG PY 2002 VL 43 IS 8 BP 1090 EP 1100 PG 11 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 582LH UT WOS:000177352100022 PM 12163636 ER PT J AU Ross, SA Poirier, L AF Ross, SA Poirier, L TI Proceedings of the trans-HHS workshop: Diet, DNA methylation processes and health SO JOURNAL OF NUTRITION LA English DT Editorial Material DE DNA methylation; S-adenosylmethionine; diet; cancer; disease ID ACID-DEFINED DIETS; CANCER RISK; FOLATE; DEFICIENCY; REDUCTASE; GENE; HYPOMETHYLATION; MUTATION; LIVERS; BREAST C1 NCI, Nutr Sci Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. US FDA, Natl Ctr Toxicol Res, Jefferson, AR 72079 USA. RP Ross, SA (reprint author), NCI, Nutr Sci Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. NR 24 TC 15 Z9 15 U1 0 U2 1 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD AUG PY 2002 VL 132 IS 8 SU S BP 2329S EP 2332S PG 4 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 583GH UT WOS:000177398500001 PM 12163686 ER PT J AU Ziegler, RG Weinstein, SJ Fears, TR AF Ziegler, RG Weinstein, SJ Fears, TR TI Nutritional and genetic inefficiencies in one-carbon metabolism and cervical cancer risk SO JOURNAL OF NUTRITION LA English DT Article; Proceedings Paper CT Trans-HHS Workshop on Diet, DNA Methylation Processes and Health CY AUG 06-08, 2001 CL BETHESDA, MARYLAND SP Natl Ctr Toxicol Res, Food & Durg Adm, Ctr Canc Res, Div Canc Prevent, NHLBI, NICHHD, NIDDK, NIEHS, Div Nutrit Res Coordinat, Off Dietary Supplements, Amer Soc Nutrit Sci, Int Life Sci Inst N Amer DE cervical cancer; epidemiology; folate; homocysteine; one-carbon metabolism ID FOLIC-ACID; SERUM CONCENTRATIONS; HOMOCYSTEINE LEVELS; METHYLMALONIC ACID; FRAGILE SITES; US WOMEN; FOLATE; DYSPLASIA; HYPOMETHYLATION; CARCINOGENESIS AB Folate deficiency has long been postulated to play a role in the etiology of cervical cancer, the third most frequent cancer among women worldwide. In a large, multiethnic community-based case-control study of invasive cervical cancer in five U.S. areas, we assessed accepted and postulated risk factors with an in-home interview and successfully obtained blood samples, at least 6 mo after completion of cancer treatment, from 51 and 68%, respectively, of interviewed cases and controls. Cases with advanced disease (6%) and/or receiving chemotherapy (4%) were excluded, leaving 183 cases and 540 controls. Serum and red blood cell folate were measured with both microbiologic and radiobinding assays. For all four folate measures, risk was moderately, but nonsignificantly, elevated for women in the lowest quartile, compared to the highest [fully adjusted relative risks (RR), including serologic human papillomavirus (HPV)-16 status = 1.2-1.6]. However, for women in the upper three homocysteine quartiles (>6.31 mumol/L), risk of invasive cervical cancer was substantially and significantly elevated (fully adjusted RR, including serologic HPV-16 status = 2.4-3.2; P for trend = 0.01). This strong relationship suggests that circulating homocysteine may be 1) an especially accurate indicator of inadequate folate, 2) an integratory measure of insufficient folate in tissues or 3) a biomarker of disruption of one-carbon metabolism. The contribution of common polymorphisms in one-carbon pathway genes, as well as inadequate vitamin B-6, vitamin B-12 and/or riboflavin, to elevated homocysteine, inefficient one-carbon metabolism and increased cervical cancer risk merits further exploration. C1 NCI, Epidemiol & Biostat Program, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Ziegler, RG (reprint author), NCI, Epidemiol & Biostat Program, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NR 30 TC 15 Z9 20 U1 0 U2 6 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD AUG PY 2002 VL 132 IS 8 SU S BP 2345S EP 2349S PG 5 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 583GH UT WOS:000177398500005 PM 12163690 ER PT J AU Srivastava, S Gopal-Srivastava, R AF Srivastava, S Gopal-Srivastava, R TI Biomarkers in cancer screening: A public health perspective SO JOURNAL OF NUTRITION LA English DT Article; Proceedings Paper CT Trans-HHS Workshop on Diet, DNA Methylation Processes and Health CY AUG 06-08, 2001 CL BETHESDA, MARYLAND SP Natl Ctr Toxicol Res, Food & Durg Adm, Ctr Canc Res, Div Canc Prevent, NHLBI, NICHHD, NIDDK, NIEHS, Div Nutrit Res Coordinat, Off Dietary Supplements, Amer Soc Nutrit Sci, Int Life Sci Inst N Amer DE biomarkers; cancer screening; early detection; public health; technology AB The last three decades have witnessed a rapid advancement and diffusion of technology in health services. Technological innovations have given health service providers the means to diagnose and treat an increasing number of illnesses, including cancer. In this effort, research on biomarkers for cancer detection and risk assessment has taken a center stage in our effort to reduce cancer deaths. For the first time, scientists have the technologies to decipher and understand these biomarkers and to apply them to earlier cancer detection. By identifying people at high risk of developing cancer, it would be possible to develop intervention efforts on prevention rather than treatment. Once fully developed and validated, then the regular clinical use of biomarkers in early detection and risk assessment will meet nationally recognized health care needs: detection of cancer at its earliest stage. The dramatic rise in health care costs in the past three decades is partly related to the proliferation of new technologies. More recent analysis indicates that technological change, such as new procedures, products and capabilities, is the primary explanation of the historical increase in expenditure. Biomarkers are the new entrants in this competing environment. Biomarkers are considered as a competing, halfway or add-on technology. Technology such as laboratory tests of biomarkers will cost less compared with computed tomography (CT) scans and other radiographs. However, biomarkers for earlier detection and risk assessment have not achieved the level of confidence required for clinical applications. This paper discusses some issues related to biomarker development, validation and quality assurance. Some data on the trends of diagnostic technologies, proteomics and genomics are presented and discussed in terms of the market share. Eventually, the use of biomarkers in health care could reduce cost by providing noninvasive, sensitive and reliable assays at a fraction of the cost of definitive technology, such as CT scan. The National Cancer Institute's Early Detection Research Network (EDRN) has begun an innovative, investigator-initiated project to improve methods for detecting the biomarkers of cancer cells. The EDRN is a consortium of more than 32 institutions to link discovery of biomarkers to the next steps in the process of developing early detection tests. These discoveries will lead to early clinical validation of tests with improved accuracy and reliability. C1 NCI, Canc Biomarkers Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. NCI, Div Extramural Activ, Bethesda, MD 20892 USA. RP Srivastava, S (reprint author), NCI, Canc Biomarkers Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. NR 16 TC 29 Z9 29 U1 0 U2 3 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD AUG PY 2002 VL 132 IS 8 SU S BP 2471S EP 2475S PG 5 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 583GH UT WOS:000177398500029 PM 12163714 ER PT J AU Mansky, PJ Liewehr, DJ Steinberg, SM Chrousos, GP Avila, NA Long, L Bernstein, D Mackall, CL Hawkins, DS Helman, LJ AF Mansky, PJ Liewehr, DJ Steinberg, SM Chrousos, GP Avila, NA Long, L Bernstein, D Mackall, CL Hawkins, DS Helman, LJ TI Treatment of metastatic osteosarcoma with the somatostatin analog OncoLar: Significant reduction of insulin-like growth factor-1 serum levels SO JOURNAL OF PEDIATRIC HEMATOLOGY ONCOLOGY LA English DT Article; Proceedings Paper CT 37th Annual Meeting of the American-Society-of-Clinical-Oncology CY MAY 12-15, 2001 CL SAN FRANCISCO, CALIFORNIA SP Amer Soc Clin Oncol DE IGF-1; osteosarcoma; OncoLar; growth hormone; IGF-1axis; phase 1 study ID I IGF-I; FACTOR BINDING-PROTEINS; NONMETASTATIC OSTEOSARCOMA; CANCER CELLS; CHEMOTHERAPY; OCTREOTIDE; EXTREMITY; TAMOXIFEN; RECEPTOR; APOPTOSIS AB Background: Insulin-like growth factor-1 (IGF-1) has been implicated in the growth and/or metastasis of osteosarcoma (OS) and chondrosarcoma based on in vitro and experimental animal studies. Study Purpose: To determine the degree of growth hormone (GH), IGF-1 axis blockade, toxicities, and antitumor effect of OncoLar (ONC) (Novartis, East Hanover, NJ, U.S.A.) in OS. Design/Methods: A phase 1 study with ONC enrolled 21 OS patients (median age 19 y) in four cohorts: ONC 60 mg or 90 mg intramuscularly every 4 weeks with/without tamoxifen (TAM) 20 mg oral daily. Results: There were no dose-limiting toxicities. Nineteen percent of patients had grade III drug-related toxicities including: 62% of patients showed progressive disease after two courses (8 wk). Nineteen percent received four courses. No clinical responses were observed. At weeks two and eight of therapy, IGF-1 serum levels dropped 46% (P < 0.0001, n = 21) and 53% (P = 0.003, n = 10). The difference of the area under the curve (AUC) minus baseline AUC (DeltaAUC) for arginine-stimulated GH serum levels at week two was lower than baseline (P < 0.01). At weeks two and eight, GH peak values were lower than baseline (P < 0.0001 and P 0.002, respectively). Conclusions: A long-acting somatostatin analog was able to lower IGF-1 levels of OS patients. IGF-BP-3 and GH were only transiently reduced. Although ONC was well tolerated, no sustained clinical responses were observed. The pathophysiology of serum versus tissue concentrations of IGF-1 as well as the interplay of IGFs, IGF-binding proteins, and other growth factors and cytokines in osteosarcoma warrants further investigation. A better understanding of these processes should lead to a more effective exploitation of these pathways for the targeted therapy of OS. C1 NCI, Pediat Oncol Branch, NIH, Bethesda, MD 20892 USA. NCI, Biostat & Data Management Sect, NIH, Bethesda, MD 20892 USA. NICHHD, NICHD, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NIH, CC, Dept Diagnost Radiol, Bethesda, MD 20892 USA. Childrens Hosp & Reg Med Ctr, Seattle, WA USA. RP Mansky, PJ (reprint author), Natl Ctr Complementary & Alternat Med, Clin Invest Lab, 8 West Dr,Quarters 15B-1,MSC2669, Bethesda, MD 20892 USA. FU NCRR NIH HHS [M01-RR-00037] NR 33 TC 32 Z9 36 U1 0 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1077-4114 J9 J PEDIAT HEMATOL ONC JI J. Pediatr. Hematol. Oncol. PD AUG-SEP PY 2002 VL 24 IS 6 BP 440 EP 446 DI 10.1097/00043426-200208000-00007 PG 7 WC Oncology; Hematology; Pediatrics SC Oncology; Hematology; Pediatrics GA 592NJ UT WOS:000177943500006 PM 12218590 ER PT J AU Wendler, D Rackoff, JE Emanuel, EJ Grady, C AF Wendler, D Rackoff, JE Emanuel, EJ Grady, C TI The ethics of paying for children's participation in research SO JOURNAL OF PEDIATRICS LA English DT Editorial Material C1 NIH, Dept Clin bioeth, Bethesda, MD 20892 USA. RP Grady, C (reprint author), NIH, Dept Clin bioeth, Bldg 10,1C118, Bethesda, MD 20892 USA. NR 7 TC 53 Z9 53 U1 0 U2 2 PU MOSBY-ELSEVIER PI NEW YORK PA 360 PARK AVENUE SOUTH, NEW YORK, NY 10010-1710 USA SN 0022-3476 EI 1097-6833 J9 J PEDIATR-US JI J. Pediatr. PD AUG PY 2002 VL 141 IS 2 BP 166 EP 171 DI 10.1067/mpd.2002.124381 PG 6 WC Pediatrics SC Pediatrics GA 586XY UT WOS:000177612500008 PM 12183709 ER PT J AU Rothbaum, R Perrault, J Vlachos, A Cipolli, M Alter, BP Burroughs, S Durie, P Elghetany, MT Grand, R Hubbard, V Rommens, J Rossi, T AF Rothbaum, R Perrault, J Vlachos, A Cipolli, M Alter, BP Burroughs, S Durie, P Elghetany, MT Grand, R Hubbard, V Rommens, J Rossi, T TI Shwachman-Diamond syndrome: Report from an international conference SO JOURNAL OF PEDIATRICS LA English DT Article ID EXOCRINE PANCREATIC DYSFUNCTION; ACUTE MYELOID-LEUKEMIA; PATIENT C1 Montreal Childrens Hosp, Montreal, PQ H3H 1P3, Canada. St Louis Childrens Hosp, St Louis, MO 63178 USA. Univ Toronto, Hosp Sick Children, Toronto, ON M5G 1X8, Canada. Schneider Childrens Hosp, New Hyde Pk, NY USA. Univ Rochester, Strong Mem Hosp, Rochester, NY 14642 USA. Cyst Fibrosis Ctr, Verona, Italy. Natl Canc Inst, Clin Genet Branch, Div Canc Epidemiol & Genet, Bethesda, MD USA. NIDDK, Nutr Sci Branch, Bethesda, MD USA. Univ Connecticut, Farmington, CT USA. Univ Texas, Med Branch, Galveston, TX 77550 USA. Childrens Hosp, Boston, MA 02115 USA. RP Perrault, J (reprint author), Montreal Childrens Hosp, 2300 Tupper, Montreal, PQ H3H 1P3, Canada. NR 33 TC 41 Z9 41 U1 0 U2 1 PU MOSBY-ELSEVIER PI NEW YORK PA 360 PARK AVENUE SOUTH, NEW YORK, NY 10010-1710 USA SN 0022-3476 EI 1097-6833 J9 J PEDIATR-US JI J. Pediatr. PD AUG PY 2002 VL 141 IS 2 BP 266 EP 270 DI 10.1067/mpd.2002.125850 PG 5 WC Pediatrics SC Pediatrics GA 586XY UT WOS:000177612500024 PM 12183725 ER PT J AU Yang, J Bagby, M Costa, PT Ryder, AG Herbst, JH AF Yang, J Bagby, M Costa, PT Ryder, AG Herbst, JH TI Assessing the DSM-IV structure of personality disorder with a sample of Chinese psychiatric patients SO JOURNAL OF PERSONALITY DISORDERS LA English DT Article ID CLUSTERS; MODELS AB The validity of the three-cluster system of personality disorders (PDs) in the latest version of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV; APA, 1994) was examined in a sample of Chinese psychiatric patients (n = 227), who completed the self-report Personality Disorders Questionnaire for DSM-IV (PDQ-4; Hyler, 1994) and who were also administered the clinician-rated Personality Disorders Interview-IV (PDI-IV; Widiger, Mangine, Corbit, Ellis, & Thomas,). Using confirmatory factor analysis, a three-factor model corresponding to the DSM-IV clusters was tested and compared statistically to a one-factor model and a set of random, three-factor models. Only the clinician-rated instrument supported the DSM-IV three-cluster model, and then only when the factors were allowed to correlate. Results from the theoretically more rigorous uncorrelated model testing did not support the DSM-IV model for either assessment modality. C1 Ctr Addict & Mental Hlth, Res Sect Personal & Psychopathol, Toronto, ON M5T 1R8, Canada. Ctr Addict & Mental Hlth, Dept Psychiat, Toronto, ON M5T 1R8, Canada. NIA, Personal Stress & Coping Sect, Lab Personal & Cognit, NIH, Bethesda, MD 20892 USA. Univ British Columbia, Dept Psychol, Vancouver, BC, Canada. RP Bagby, M (reprint author), Ctr Addict & Mental Hlth, Clin Res Dept, Clarke Site,250 Coll St, Toronto, ON M5T 1R8, Canada. RI Ryder, Andrew/A-1108-2008; OI Ryder, Andrew/0000-0003-3041-7168; Costa, Paul/0000-0003-4375-1712 NR 30 TC 26 Z9 27 U1 0 U2 2 PU GUILFORD PUBLICATIONS INC PI NEW YORK PA 72 SPRING STREET, NEW YORK, NY 10012 USA SN 0885-579X J9 J PERS DISORD JI J. Pers. Disord. PD AUG PY 2002 VL 16 IS 4 BP 317 EP 331 DI 10.1521/pedi.16.4.317.24127 PG 15 WC Psychiatry SC Psychiatry GA 589JG UT WOS:000177754700003 PM 12224125 ER PT J AU Weiss, R Mevissen, M Hauser, DS Scholtysik, G Portier, CJ Walter, B Studer, UE Danuser, H AF Weiss, R Mevissen, M Hauser, DS Scholtysik, G Portier, CJ Walter, B Studer, UE Danuser, H TI Inhibition of human and pig ureter motility in vitro and in vivo by the K+ channel openers PKF 217-744b and nicorandil SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID SENSITIVE POTASSIUM CHANNELS; NITRIC-OXIDE; SMOOTH-MUSCLE; PERISTALTIC FUNCTION; INTRAVESICAL URETER; MESENTERIC-ARTERIES; CYCLIC-GMP; GLIBENCLAMIDE; RECEPTOR; CROMAKALIM AB The relaxing property of the K+ channel opener and nitric oxide donor nicorandil and the new K+ channel opener PKF 217-744b was investigated on isolated human ureteral tissue in vitro and in intact ureters of anesthetized pigs in vivo. In addition, nicorandil and its antagonists, glibenclamide and methylene blue, were tested on isolated pig ureter tissue in vitro. Nicorandil decreased the frequency of spontaneous contractions in isolated pig ureter rings. This effect was antagonized by glibenclamide and methylene blue suggesting that the nicorandil induced relaxation of the ureter is mediated by activation of ATP-sensitive K+ channels and involvement of soluble guanylate cyclase. Moreover, nicorandil and PKF 217-744b reduced the amplitude of electrically induced contractions in isolated human ureter rings. Calculations of EC50 values showed that PKF 217-744b [EC50 = 4.83 x 10(-8) M] was more potent than nicorandil [EC50 = 4.38 x 10(-5) M]. Both drugs reduced the contraction frequency of the pig ureter after intravenous and topical administration in vivo. Intravenous, but not topical, administration of nicorandil and PKF 217-744b significantly decreased arterial blood pressure but did not affect the heart rate. The in vitro findings suggest that K+ channel opening and nitric oxide release mediate the effect of nicorandil. Our in vivo results indicate that PKF 217- 744b and nicorandil are promising drugs for clinical application in patients with acute stone colic to relieve obstruction and facilitate stone passage or to relax the ureter before ureteroscopy. C1 Univ Bern, Inst Vet Pharmacol, Bern, Switzerland. Univ Bern, Dept Urol, Bern, Switzerland. NIEHS, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. RP Danuser, H (reprint author), Univ Bern, Inselspital, Dept Urol, CH-3010 Bern, Switzerland. RI Portier, Christopher/A-3160-2010 OI Portier, Christopher/0000-0002-0954-0279 NR 35 TC 19 Z9 20 U1 0 U2 2 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD AUG PY 2002 VL 302 IS 2 BP 651 EP 658 AR UNSP 4787/995633 DI 10.1124/jpet.302.2.651 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 574PR UT WOS:000176899200032 PM 12130728 ER PT J AU Gould, TD Manji, HK AF Gould, TD Manji, HK TI Signaling networks in the pathophysiology and treatment of mood disorders SO JOURNAL OF PSYCHOSOMATIC RESEARCH LA English DT Review DE adenylate cyclase; bipolar disorder; depression; G proteins; phosphotidylinositol; second messenger systems ID PROTEIN-KINASE-C; BIPOLAR AFFECTIVE-DISORDER; ADENYLATE-CYCLASE ACTIVITY; CHRONIC ANTIDEPRESSANT TREATMENT; RAT CEREBRAL-CORTEX; NUCLEOTIDE-BINDING PROTEINS; BETA-ADRENERGIC RECEPTORS; LONG-TERM LITHIUM; MANIC-DEPRESSIVE ILLNESS; CYCLIC-AMP PRODUCTION AB Over the past decade, the focus of research into the pathophysiology of mood disorders (bipolar disorder and unipolar depression in particular) has shifted from an interest in the biogenic amines to an emphasis on second messenger systems within cells. Second messenger systems rely on cell membrane receptors to relay information from the extracellular environment to the interior of the cell. Within the cell, this information is processed and altered, eventually to the point where gene and protein expression patterns are changed. There is a preponderance of evidence implicating second messenger systems and their primary contact with the extracellular environment, G proteins, in the pathophysiology of mood disorders. After an introduction to G proteins and second messenger pathways, this review focuses on the evidence implicating G proteins and two second messenger systems-the adenylate cyclase (cyclic adenosine monophosphate, cAMP) and phosphoinositide (protein kinase C, PKC) intracellular signaling cascades-in the pathophysiologgy and treatment of bipolar disorder and unipolar depression. Emerging evidence implicates changes in cellular resiliency, neuroplasticity and additional cellular pathways in the pathophysiology of mood disorders. The systems discussed within this review have been implicated in neuroplastic processes and in modulation of many other cellular pathways, making them likely candidates for mediators of these findings. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NIMH, Mol Pathophysiol Lab, NIH, Bethesda, MD 20892 USA. RP Manji, HK (reprint author), NIMH, Mol Pathophysiol Lab, NIH, Bldg 49,Room B1EE16, Bethesda, MD 20892 USA. NR 129 TC 78 Z9 83 U1 1 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0022-3999 J9 J PSYCHOSOM RES JI J. Psychosomat. Res. PD AUG PY 2002 VL 53 IS 2 BP 687 EP 697 AR PII S0022-3999(02)00426-9 DI 10.1016/S0022-3999(02)00426-9 PG 11 WC Psychiatry SC Psychiatry GA 588KN UT WOS:000177700800008 PM 12169343 ER PT J AU Berry, JM Jobe, JB AF Berry, JM Jobe, JB TI At the intersection of personality and adult development SO JOURNAL OF RESEARCH IN PERSONALITY LA English DT Editorial Material AB People change and remain the same over time. This assertion is simple but not simplistic. Two decades of dogged research on the change-stability debate have finally led proponents on both sides to cede: "It's both." Meta-analyses of personality research (Roberts & DelVecchio, 2000) reveal that different statistical indices provide different answers to the change-stability question: rank-order correlation coefficients support stability, group and individual mean level indices support change. Yet how much change occurs, and when? For which traits and states? What is the relevance of change versus stability for various outcomes? Although trait structure and rank-order position may change little to not at all over time, are the manifestations of traits and their associations with daily activities and life events as stable? Who is more likely to change, and how does change at the fundamental level of personality affect other life domains, such as health, emotion, relationships, leisure, and happiness? This special issue of papers provides empirical evidence couched in theoretical perspectives that addresses most of the questions posed above. Of those not yet answered, promising paradigms for getting those answers are proffered for the new and continuing generations of behavioral scientists who study personality as a life-long process. (C) 2002 Published by Elsevier Science (USA). C1 Univ Richmond, Dept Psychol, Richmond, VA 23173 USA. NHLBI, Richmond, VA USA. RP Berry, JM (reprint author), Univ Richmond, Dept Psychol, Richmond, VA 23173 USA. EM jberry@richmond.edu NR 6 TC 1 Z9 1 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0092-6566 J9 J RES PERS JI J. Res. Pers. PD AUG PY 2002 VL 36 IS 4 BP 283 EP 286 AR PII S0092-6566(02)00009-0 DI 10.1016/S0092-6566(02)00009-0 PG 4 WC Psychology, Social SC Psychology GA 582KA UT WOS:000177348400001 ER PT J AU McCrae, RR AF McCrae, RR TI The maturation of personality psychology: Adult personality development and psychological well-being SO JOURNAL OF RESEARCH IN PERSONALITY LA English DT Article ID AGE-DIFFERENCES; LIFE-SPAN; STABILITY; TRAITS; NEUROTICISM; HAPPINESS; HAPPY AB Personality psychology has made striking advances in the past two decades, demonstrating the importance of individual differences in a wide variety of life domains. Longitudinal studies of adult development contributed to these advances by revealing the stability of personality traits even in the face of changing life circumstances. More recently, cross-culturally replicated patterns of adult age differences have suggested that traits are endogenous dispositions with intrinsic paths of development. The importance of personality traits was underscored by research linking dimensions of personality to psychological well-being and mental health, and the rapidly maturing science of personality psychology holds promise for understanding many other social and psychological phenomena. C1 NIA, Lab Personal & Cognit, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. RP McCrae, RR (reprint author), NIA, Lab Personal & Cognit, Gerontol Res Ctr, NIH, Box 03,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 58 TC 28 Z9 28 U1 1 U2 3 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0092-6566 J9 J RES PERS JI J. Res. Pers. PD AUG PY 2002 VL 36 IS 4 BP 307 EP 317 AR PII S0092-6566(02)00011-9 DI 10.1016/S0092-6566(02)00011-9 PG 11 WC Psychology, Social SC Psychology GA 582KA UT WOS:000177348400003 ER PT J AU Kashdan, TB Pelham, WE Lang, AR Hoza, B Jacob, RG Jennings, JR Blumenthal, JD Gnagy, EM AF Kashdan, TB Pelham, WE Lang, AR Hoza, B Jacob, RG Jennings, JR Blumenthal, JD Gnagy, EM TI Hope and optimism as human strengths in parents of children with externalizing disorders: Stress is in the eye of the beholder SO JOURNAL OF SOCIAL AND CLINICAL PSYCHOLOGY LA English DT Article ID ATTENTION-DEFICIT DISORDER; PSYCHOSOCIAL TREATMENTS; ALCOHOL-CONSUMPTION; BEHAVIOR PROBLEMS; SOCIAL SUPPORT; LIFE STRESS; PERCEPTIONS; ADJUSTMENT; DEPRESSION; SYMPTOMS AB We examined hope as a potential resiliency factor for the daily strains of raising children with disruptive behavior disorders. In light of the motivational component of hope theory, initiating and sustaining effort toward goals (i.e., agency), we were interested in hope's relation to constructs addressing self-esteem, familial functioning, and stress. Two hundred, fifty-two parents of children with externalizing disorders completed self-report questionnaires. Significant associations were foundamong hope and parental and familial functioning indices (e.g., warm and nurturing parenting styles, cohesive and active family environment, adaptive coping strategies). Considering their conceptual overlap, we tested the unique predictive power of hope and optimistic attributions on indices of psychological functioning. Separate regressions indicated that hope significantly predicted psychological functioning beyond what was accounted for by social desirability, the severity of child symptoms, and optimistic attributions. Hope agency compared to hope pathways (i.e., perceived ability to generate strategies to obtain goals) accounted for the vast amount of variance in regression models. In contrast, optimistic attributions failed to predict any of the variables of interest. Treatment and prevention strategies are suggested with an integrated focus on both the disruptive behaviors of children and parental character traits. C1 SUNY Buffalo, Dept Psychol, Buffalo, NY 14260 USA. Florida State Univ, Tallahassee, FL 32306 USA. Purdue Univ, W Lafayette, IN 47907 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. NIMH, Bethesda, MD 20892 USA. RP Kashdan, TB (reprint author), SUNY Buffalo, Dept Psychol, Pk Hall,Box 604110, Buffalo, NY 14260 USA. NR 77 TC 26 Z9 26 U1 5 U2 17 PU GUILFORD PUBLICATIONS INC PI NEW YORK PA 72 SPRING STREET, NEW YORK, NY 10012 USA SN 0736-7236 J9 J SOC CLIN PSYCHOL JI J. Soc. Clin. Psychol. PD AUG PY 2002 VL 21 IS 4 BP 441 EP 468 DI 10.1521/jscp.21.4.441.22597 PG 28 WC Psychology, Clinical; Psychology, Social SC Psychology GA 586RY UT WOS:000177599900006 ER PT J AU Deroo, BJ Archer, TK AF Deroo, BJ Archer, TK TI Differential activation of the I kappa B alpha and mouse mammary tumor virus promoters by progesterone and glucocorticoid receptors SO JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY LA English DT Review DE progestin; glucocorticoid; MMTV; promoter; I-kappa-B ID TRANSCRIPTIONAL ACTIVATION; CHROMATIN STRUCTURE; RESPONSE ELEMENTS; HORMONE RECEPTORS; GENE ACTIVATION; DOWN-REGULATION; PROGESTINS; EXPRESSION; ANDROGEN; SEQUENCE AB The glucocorticoid and progesterone receptors (GR and PR) are structurally homologous and bind a common hormone response element (HRE). The mechanisms by which receptors activate specific promoters when multiple steroids are present in a cell is a critical question in endocrinology. To investigate how co-existing steroid receptors regulate gene transcription, we have compared two hormone-responsive promoters in T47D/A1-2 human breast cancer cells expressing both the GR and PR. The promoters chosen were those for the mouse mammary tumor virus (MMTV) and the gene for IkappaBalpha, the inhibitor of the ubiquitous transcription factor, nuclear factor kappa B (NFkappaB). Several differences between glucocorticoid and progestin activation of the IkappaBalpha and MMTV promoters were revealed. Both steroids activated the endogenous IkappaBalpha promoter, while only glucocorticoids activated a stably integrated MMTV promoter. In combination, progestins enhanced glucocorticoid activation of IkappaBalpha, but antagonized glucocorticoid activation of MMTV. These differences in steroid receptor competition were further demonstrated when levels of the PR were reduced by prolonged treatment with progestin. Under these conditions, the PR no longer competes effectively with the GR for activation of the MMTV promoter. However, on the IkappaBalpha promoter, the GR and PR still activate the promoter in a cooperative fashion. Another difference between the two promoters is their chromatin structure. In this cell line, the MMTV promoter chromatin is "closed" and insensitive to restriction enzyme cleavage, while the IkappaBalpha promoter is "open." Using PR antagonists, we demonstrate that at least one cofactor complex, the BRG-1 chromatin remodeling complex, differentially contributes to activation of both promoters. Published by Elsevier Science Ltd. C1 NIEHS, Chromat & Gene Express Sect, NIH, Res Triangle Pk, NC 27709 USA. Univ Western Ontario, Dept Biochem, London, ON N6A 4L6, Canada. RP Archer, TK (reprint author), NIEHS, Chromat & Gene Express Sect, NIH, 111 Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 36 TC 16 Z9 16 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-0760 J9 J STEROID BIOCHEM JI J. Steroid Biochem. Mol. Biol. PD AUG PY 2002 VL 81 IS 4-5 BP 309 EP 317 AR PII S0960-0760(02)00072-9 DI 10.1016/S0960-0760(02)00072-9 PG 9 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 611NC UT WOS:000179023100002 PM 12361720 ER PT J AU Rubenstein, DS Blauvelt, A Chen, SC Darling, TN AF Rubenstein, DS Blauvelt, A Chen, SC Darling, TN TI The future of academic dermatology in the United States: Report on the resident retreat for future physician-scientists, June 15-17, 2001 SO JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY LA English DT Article AB Fewer and fewer residents appear to be choosing careers in academic dermatology and research. Recognizing the impact that this trend would have on the future of our specialty, the Society for Investigative Dermatology invited a group of dermatology residents and academic dermatologists to participate in a retreat whose aim was to identify the reasons behind this trend and to begin to address these issues. C1 Univ N Carolina, Sch Med, Dept Dermatol, Chapel Hill, NC 27599 USA. NCI, Dermatol Branch, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Dept Dermatol, Atlanta, GA 30322 USA. Uniformed Serv Univ Hlth Sci, Dept Dermatol, Bethesda, MD 20814 USA. RP Rubenstein, DS (reprint author), Univ N Carolina, Sch Med, Dept Dermatol, Suite 1300,Thurston Bowles Bldg,CB 7287, Chapel Hill, NC 27599 USA. OI Darling, Thomas/0000-0002-5161-1974 NR 2 TC 17 Z9 17 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0190-9622 J9 J AM ACAD DERMATOL JI J. Am. Acad. Dermatol. PD AUG PY 2002 VL 47 IS 2 BP 300 EP 303 DI 10.1067/mjd.2002.123491 PG 4 WC Dermatology SC Dermatology GA 579YJ UT WOS:000177206300017 PM 12140479 ER PT J AU Beenken, SW Hockett, R Grizzle, W Weiss, HL Pickens, A Perloff, M Malone, WF Bland, KI AF Beenken, SW Hockett, R Grizzle, W Weiss, HL Pickens, A Perloff, M Malone, WF Bland, KI TI Transforming growth factor-alpha: A surrogate endpoint biomarker? SO JOURNAL OF THE AMERICAN COLLEGE OF SURGEONS LA English DT Article; Proceedings Paper CT 6th International Symposium on Predictive Oncology and Intervention Strategies CY FEB, 2002 CL PARIS, FRANCE ID FACTOR RECEPTOR GENE; SQUAMOUS-CELL CARCINOMAS; TGF-ALPHA; RET PROTOONCOGENE; ORAL LEUKOPLAKIA; MESSENGER-RNAS; RETINOIC ACID; HUMAN-BREAST; PREMALIGNANT LESIONS; NECK TUMORIGENESIS AB BACKGROUND: Dysplastic oral leukoplakia. (DOL) has been the index lesion in prevention trials for upper aerodigestive tract squamous cell carcinoma (SCC). Vitamin A derivatives, including 13-cis retinoic acid (13-CRA), have been used to treat DOL and to reduce the risk of subsequent SCC. Results from a trial of 13-CRA in patients with DOL are presented here. Transforming growth factor-alpha (TGF-alpha) and the epidermal growth factor receptor messenger RNA (mRNA) expression were studied to validate their use as surrogate endpoint biomarkers in prevention trials for SCC. STUDY DESIGN: In a prospective, randomized, double-blind trial of 13-CRA in 28 patients with DOL, TGF-alpha and epidermal growth factor receptor mRNA expression were analyzed in sequential biopsy specimens of DOL and of adjacent normal-appearing mucosa, utilizing a quantitative, competitive, reverse transcriptase polymerase chain reaction and were compared using the Wilcoxon signed-rank test for paired comparisons. RESULTS: In biopsy specimens of DOL, TGF-alpha mRNA expression at baseline, but not baseline expression of epidermal growth factor receptor mRNA, was significantly elevated when compared with its expression in specimens from adjacent normal-appearing mucosa (p = 0.003). In patients randomized to 13-CRA who had greater than or equal to 50% clearance of DOL during treatment, significant modulation of TGF-alpha mRNA overexpression was seen after 6 months of treatment (p = 0.016). TGF-alpha mRNA overexpression at baseline predicted a subsequent response to 13-CRA (p 0.066). CONCLUSIONS: The full extent of the association between TGF-alpha overexpression and the development of SCC is unknown. Evidence is presented in this article that TGF-alpha overexpression mediates the relationship between 13-CRA and DOL, but there is no direct evidence that it mediates the relationship between 13-CRA and the prevention of SCC. Determination of the extent to which TGF-alpha overexpression mediates this relationship and complete validation of TGF-alpha's role as a surrogate endpoint biomarker await the results of animal and human trials that utilize reduction in the incidence of SCC as their endpoint. (C) 2002 by the American College of Surgeons. C1 Univ Alabama, Dept Surg, Birmingham, W Midlands, England. Univ Alabama, Dept Pathol, Birmingham, W Midlands, England. Univ Alabama, Biostat Unit, Ctr Comprehens Canc, Birmingham, W Midlands, England. Eli Lilly & Co, Indianapolis, IN 46285 USA. NCI, Div Chemoprevent, Bethesda, MD 20892 USA. RP Beenken, SW (reprint author), Univ Alabama, Wallace Tumour Inst 620, 1824 6th Ave S, Birmingham, W Midlands, England. FU NCI NIH HHS [5 U01 CA68639-03] NR 53 TC 8 Z9 9 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1072-7515 J9 J AM COLL SURGEONS JI J. Am. Coll. Surg. PD AUG PY 2002 VL 195 IS 2 BP 149 EP 158 AR PII S1072-7515(02)01219-X DI 10.1016/S1072-7515(02)01219-X PG 10 WC Surgery SC Surgery GA 579GY UT WOS:000177171300001 PM 12168960 ER PT J AU Horowitz, AM Canto, MT Child, WL AF Horowitz, AM Canto, MT Child, WL TI Maryland adults' perspectives on oral cancer prevention and early detection SO JOURNAL OF THE AMERICAN DENTAL ASSOCIATION LA English DT Article ID KNOWLEDGE AB Overview. In any given 12-month period, Maryland ranks 27th among all states and the District of Columbia in estimated new cases of oral cancer The state also has the seventh highest overall mortality rate for oral cancer. Because of earlier research indicating that Maryland adults had little knowledge and many misconceptions about oral cancer, the authors undertook a study to obtain in-depth information from Maryland adults 40 years of age or older on oral cancer, oral cancer examinations and factors associated with having an oral cancer examination. Methods. The authors conducted a qualitative descriptive study using information gathered from three focus groups consisting of nine, 10 and seven adults respectively, and which met at two locations. The authors hired a private focus group research firm, which randomly selected participants from a telephone list of local residents. A professionally trained moderator conducted all focus groups using a semistructured interview guide. Results. Participants were struck by the fact that they rarely hear about this type of cancer. Many said that they never had had an oral cancer examination and did not know there was such a thing. Many participants also reported that they likely would be more comfortable discussing oral cancer with their physicians than with their dentists. Conclusions. These findings provide additional in depth insights to earlier work about Maryland adults oral cancer knowledge, opinions and practices. The state plans to use this information to develop educational materials and interventions for the public to promote oral cancer prevention and early detection in Maryland. Clinical Implications. Extensive public education about oral and pharyngeal cancers should be provided in dental offices and clinics as well as in mass media of all types. More clinicians should include comprehensive oral cancer screenings in their oral examinations, and they should explain to patients what they are doing when they provide these screenings. C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD 20892 USA. RP Horowitz, AM (reprint author), Natl Inst Dent & Craniofacial Res, NIH, Bldg 45,Room 3AN-44B, Bethesda, MD 20892 USA. NR 14 TC 29 Z9 29 U1 1 U2 6 PU AMER DENTAL ASSN PI CHICAGO PA 211 E CHICAGO AVE, CHICAGO, IL 60611 USA SN 0002-8177 J9 J AM DENT ASSOC JI J. Am. Dent. Assoc. PD AUG PY 2002 VL 133 IS 8 BP 1058 EP 1063 PG 6 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 585GA UT WOS:000177516000016 PM 12198984 ER PT J AU Greger, JL Maly, A Jensen, N Kuhn, J Monson, K Stocks, A AF Greger, JL Maly, A Jensen, N Kuhn, J Monson, K Stocks, A TI Food pantries can provide nutritionally adequate food packets but need help to become effective referral units for public assistance programs SO JOURNAL OF THE AMERICAN DIETETIC ASSOCIATION LA English DT Article ID NUTRIENT INTAKE; HOUSEHOLDS C1 Univ Wisconsin, Dept Nutr Sci, Madison, WI 53706 USA. NIH, Bethesda, MD 20892 USA. Univ Wisconsin, Dept Nutr Sci, Madison, WI 53706 USA. RP Greger, JL (reprint author), Univ Connecticut, 438 Whitney Rd,Ext,Unit 1006, Storrs, CT 06269 USA. NR 12 TC 4 Z9 4 U1 2 U2 8 PU AMER DIETETIC ASSOC PI CHICAGO PA 216 W JACKSON BLVD #800, CHICAGO, IL 60606-6995 USA SN 0002-8223 J9 J AM DIET ASSOC JI J. Am. Diet. Assoc. PD AUG PY 2002 VL 102 IS 8 BP 1126 EP 1128 DI 10.1016/S0002-8223(02)90249-X PG 3 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 581JE UT WOS:000177287800017 PM 12171458 ER PT J AU Kleta, R Gahl, WA AF Kleta, R Gahl, WA TI Cystinosis: Antibodies and healthy bodies SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Editorial Material ID NEPHROPATHIC CYSTINOSIS; GRANULAR FRACTIONS; COUNTER-TRANSPORT; CORNEAL CRYSTALS; GENE CTNS; CYSTEAMINE; FIBROBLASTS; MUTATIONS; LYSOSOMES; DELETION C1 NICHHD, Sect Human Biochem Genet, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Gahl, WA (reprint author), NICHD, NIH, 10 Ctr Dr,MSC 1830,Bldg 10,Room 9S-241, Bethesda, MD 20892 USA. NR 44 TC 9 Z9 10 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1046-6673 J9 J AM SOC NEPHROL JI J. Am. Soc. Nephrol. PD AUG PY 2002 VL 13 IS 8 AR UNSP 1046-6673/1308-2189 DI 10.1097/01.ASN.0000027098.90648.F2 PG 3 WC Urology & Nephrology SC Urology & Nephrology GA 575RE UT WOS:000176961000033 PM 12138155 ER PT J AU Altekruse, SF Elvinger, F Lee, KY Tollefson, LK Pierson, FW Eifert, J Sriranganathan, N AF Altekruse, SF Elvinger, F Lee, KY Tollefson, LK Pierson, FW Eifert, J Sriranganathan, N TI Antimicrobial susceptibilities of Escherichia coli strains from a turkey operation SO JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION LA English DT Article ID ANTIBIOTIC-RESISTANCE; SALMONELLA; PREVALENCE; CHICKENS; MEAT AB Objective-To measure minimum inhibitory concentrations (MIC) of 17 antimicrobials for Escherichia coli isolates from a turkey operation and assess whether small samples provide precise estimates of geometric mean MIC. Design-Prospective study. Sample Population-105 clinical isolates from birds and 1,104 fecal isolates from 20 flocks (poults and finisher hens). Procedure-A Mueller-Hinton broth dilution panel was used to measure MIC, and MIC of fecal and clinical isolates were compared. We drew random samples of 5, 10, 15, 20, 25, 30, 35, 40, and 45 isolates from each finisher flock and between 100 and 105 isolates from 5, 7, 10, and 20 flocks. Antimicrobial usage was determined for enrolled flocks. Results-Six of 12 poult and 18 of 20 finisher flocks had been treated with antimicrobials, often for respiratory illnesses consistent with colibacillosis. All birds received gentamicin at the hatchery. More fecal than clinical isolates were resistant to ampicillin; however, more clinical isolates were resistant to ciprofloxacin, gentamicin, and sulfamethoxazole. Precise estimates of geometric mean MIC for flocks were obtained when greater than or equal to 15 fecal isolates were obtained per flock and, for the operation, when 105 isolates were obtained from 7 flocks. Conclusion and Clinical Relevance-Antimicrobial usage was common and may have contributed to the resistance patterns of isolates. With a modest allocation of laboratory resources, producers can monitor antimicrobial susceptibilities of clinical and fecal E coli to manage risks of antimicrobial usage and resistance. C1 Virginia Maryland Reg Coll Vet Med, Dept Large Anim Clin Sci, Blacksburg, VA 24061 USA. Virginia Maryland Reg Coll Vet Med, Dept Biomed Sci & Pathobiol, Blacksburg, VA 24061 USA. Virginia Polytech Inst & State Univ, Coll Agr & Life Sci, Dept Food Sci & Technol, Blacksburg, VA 24061 USA. US FDA, Ctr Vet Med, Rockville, MD 20895 USA. RP Altekruse, SF (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,Room 7082 MSC 7234, Rockville, MD 20852 USA. NR 26 TC 19 Z9 21 U1 0 U2 4 PU AMER VETERINARY MEDICAL ASSOC PI SCHAUMBURG PA 1931 N MEACHAM RD SUITE 100, SCHAUMBURG, IL 60173-4360 USA SN 0003-1488 J9 J AM VET MED ASSOC JI J. Am. Vet. Med. Assoc. PD AUG 1 PY 2002 VL 221 IS 3 BP 411 EP 416 DI 10.2460/javma.2002.221.411 PG 6 WC Veterinary Sciences SC Veterinary Sciences GA 577JU UT WOS:000177058400016 PM 12164541 ER PT J AU Hostetter, TH Lising, M AF Hostetter, TH Lising, M TI National Kidney Disease Education Program SO JOURNAL OF THE NATIONAL MEDICAL ASSOCIATION LA English DT Article DE burden of disease; minority populations; education program; end-stage renal disease ID RENAL-DISEASE; PREVENTION AB The predicted increase in the number of people with end-stage renal disease (ESRD) places an enormous burden on healthcare providers and the Medicare system. Recognizing this problem led the Council of American Kidney Societies to conceive, develop, and implement the National Kidney Disease Education, Program (NKDEP). It was decided to aim the program at both patients and primary care providers in an attempt to halt the relentless increase in the numbers of patients with ESRD. The development of straightforward targets for therapy and screening are a goal of the NKDEP. The strategic plan of the program calls for all those at high risk of developing ESRD to seek testing. Working groups are currently addressing the components of the plan and are actively developing messages and guidance for target audiences. The NKDEP aims to reduce the incidence of ESRD. Even though the results may not come quickly, it is hoped that ESRD mortality can be reduced over the coming years. C1 NIDDK, Natl Kidney Dis Educ Program, NIH, Bethesda, MD 20892 USA. RP Hostetter, TH (reprint author), NIDDK, Natl Kidney Dis Educ Program, NIH, 6707 Democracy Blvd,Room 645, Bethesda, MD 20892 USA. NR 11 TC 4 Z9 5 U1 0 U2 0 PU NATL MED ASSOC PI WASHINGON PA 1012 10TH ST, N W, WASHINGON, DC 20001 USA SN 0027-9684 J9 J NATL MED ASSOC JI J. Natl. Med. Assoc. PD AUG PY 2002 VL 94 IS 8 SU S BP 72S EP 75S PG 4 WC Medicine, General & Internal SC General & Internal Medicine GA 578QH UT WOS:000177129000010 PM 12152915 ER PT J AU Kimmel, PL Patel, SS Peterson, RA AF Kimmel, PL Patel, SS Peterson, RA TI Depression in African-American patients with kidney disease SO JOURNAL OF THE NATIONAL MEDICAL ASSOCIATION LA English DT Article ID STAGE RENAL-DISEASE; URBAN HEMODIALYSIS-PATIENTS; CHRONIC DIALYSIS PATIENTS; LONG-TERM DIALYSIS; MAJOR DEPRESSION; UNITED-STATES; PRIMARY-CARE; BEHAVIORAL COMPLIANCE; PSYCHOSOCIAL FACTORS; GLOBAL BURDEN AB There are few data on the epidemiology consequences and treatment of depression in African-American patients with kidney disease in the US, even though such patients disproportionately bear the burden of this illness. This paper reviews data on the diagnosis and pathogenesis of depression and its consequences in patients with and without kidney disease, in addition to work on the epidemiology of depression in the African-American population and in the US End-stage Renal Disease (ESRD) program. African Americans are thought to have similar susceptibility to the development of depression as other populations in the US, but diminished access to care for this group of patients may be associated with differential outcomes. Data are presented from longitudinal studies of psychosocial outcomes in a population comprising primarily African-American patients with ESRD, and is reviewed the treatment of depression in patients with and without kidney disease. There are few studies of the management of depression that focus on minority populations. The authors agree with recommendations that treatment trials should include minority patients, patients with medical comorbidities, and the elderly and assess function and quality of life as outcomes. The relationships between age, marital status and satisfaction, ethnicity, and perception of quality of life and depressive affect level and diagnosis of depression, and medical outcomes have not been determined in ESRD patients, or in African-American patients with ESRD. There are few studies of drugs for the treatment of depression in ESRD patients, and only one small randomized controlled trial. These have shown that therapy with selective serotonin reuptake inhibitors appears to be a safe treatment option for patients with ESRD. The long-term effectiveness of therapy, and its association with clinically important outcomes such as perception of quality of life, compliance, and survival have not been evaluated in ESRD patients. Also, therapeutic effectiveness and outcomes have not been assessed in minority populations with ESRD. These issues need to be addressed to optimize the management of depression in African Americans with kidney disease. C1 NIDDKD, Div Kidney Urol & Hematol Dis, NIH, Bethesda, MD 20892 USA. RP Kimmel, PL (reprint author), NIDDKD, Div Kidney Urol & Hematol Dis, NIH, 6707 Democracy Blvd, Bethesda, MD 20892 USA. EM kimmelp@extra.niddk.nih.gov NR 80 TC 15 Z9 15 U1 1 U2 3 PU NATL MED ASSOC PI WASHINGON PA 1012 10TH ST, N W, WASHINGON, DC 20001 USA SN 0027-9684 J9 J NATL MED ASSOC JI J. Natl. Med. Assoc. PD AUG PY 2002 VL 94 IS 8 SU S BP 92S EP 103S PG 12 WC Medicine, General & Internal SC General & Internal Medicine GA 578QH UT WOS:000177129000013 PM 12152919 ER PT J AU Macedonia, C Miller, JL Sonies, BC AF Macedonia, C Miller, JL Sonies, BC TI Power Doppler imaging of the fetal upper aerodigestive tract using a 4-point standardized evaluation - Preliminary report SO JOURNAL OF ULTRASOUND IN MEDICINE LA English DT Article DE fetal activity; fetal ingestion; fetal respiration; fetal swallowing; fetal upper aerodigestive tract; power Doppler imaging ID BRACHMANN-DELANGE-SYNDROME; PRENATAL-DIAGNOSIS; BIRTH-WEIGHT; UPPER AIRWAY; ATRESIA; FLOW; DYSFUNCTION; MANAGEMENT; SURVIVAL; INFANTS AB Objective. Our investigation of fetal swallowing has identified potential limitations in the use of color Doppler imaging for detection of amniotic fluid flow and discrimination of respiratory from ingestive activity. The objective of this study was to evaluate an alternative imaging modality, power Doppler sonography, as a technique to enhance detection of amniotic fluid flow in the upper aerodigestive tract. Methods. We applied a standardized 4-axis sonographic examination of upper aerodigestive structures and used power Doppler imaging to document amniotic fluid flow. Normal aerodigestive activities from 62 healthy control subjects were compared with 4 abnormal cases. Results. Our longitudinal experience with 66 subjects showed that a directed evaluation of the fetal upper aerodigestive tract with power Doppler imaging provided a systematic approach for studying the physiologic development of this region in both healthy and at-risk fetuses. Conclusions. A standardized 4-axis examination with power Doppler imaging is a useful adjunct in addressing ingestive and respiratory functions in the developing fetus. C1 Uniformed Serv Univ Hlth Sci, Natl Naval Med Ctr, Dept Obstet & Gynecol, Bethesda, MD 20814 USA. NIH, Bethesda, MD 20892 USA. RP Macedonia, C (reprint author), Uniformed Serv Univ Hlth Sci, Natl Naval Med Ctr, Dept Obstet & Gynecol, 4301 Jones Bridge Rd, Bethesda, MD 20814 USA. NR 28 TC 7 Z9 8 U1 0 U2 0 PU AMER INST ULTRASOUND MEDICINE PI LAUREL PA SUBSCRIPTION DEPT, 14750 SWEITZER LANE, STE 100, LAUREL, MD 20707-5906 USA SN 0278-4297 J9 J ULTRAS MED JI J. Ultrasound Med. PD AUG PY 2002 VL 21 IS 8 BP 869 EP 878 PG 10 WC Acoustics; Radiology, Nuclear Medicine & Medical Imaging SC Acoustics; Radiology, Nuclear Medicine & Medical Imaging GA 581HQ UT WOS:000177286200009 PM 12164572 ER PT J AU Schaeffer, AJ Landis, JR Knauss, JS Propert, KJ Alexander, RB Litwin, MS Nickel, JC O'Leary, MP Nadler, RB Pontari, MA Shoskes, DA Zeitlin, SI Fowler, JE Mazurick, CA Kishel, L Kusek, JW Nyberg, LM AF Schaeffer, AJ Landis, JR Knauss, JS Propert, KJ Alexander, RB Litwin, MS Nickel, JC O'Leary, MP Nadler, RB Pontari, MA Shoskes, DA Zeitlin, SI Fowler, JE Mazurick, CA Kishel, L Kusek, JW Nyberg, LM CA Chronic Prostatitis Collaborative TI Demographic and clinical characteristics of men with chronic prostatitis: The National Institutes of Health Chronic Prostatitis Cohort study SO JOURNAL OF UROLOGY LA English DT Article DE prostate; prostatitis; questionnaires; pelvic pain; demography ID SYMPTOM INDEX; COMBINATION; ANTIBIOTICS AB Purpose: We describe the study design of the National Institutes of Health Chronic Prostatitis Cohort (CPC) study characterizing men with chronic prostatitis/the chronic pelvic pain syndrome. Materials and Methods: All 488 men screened into the CPC study before close of recruitment on August 22, 2001 were selected for analysis. The National Institutes of Health Chronic Prostatitis Symptom Index, including subscores, was used to measure symptoms. A comprehensive history, physical examination and demographic profile were obtained from each participant. Generalized Mantel-Haenszel procedures were used to investigate baseline associations between selected factors and symptoms. Results: Chronic prostatitis/chronic pelvic pain syndrome is a chronic syndrome affecting men over a wide age range. The majority of CPC study participants are white, well educated and affluent. However, lower education, lower income and unemployment were associated with more severe symptoms. Patients most frequently reported pain in the perineum and tenderness in the prostate. The highest self-reported diseases were genitourinary (55%), allergies (53%), neurological (40%) and hematopoietic, lymphatic or infectious (40%). This disease has a significant negative impact on mental and physical domains of quality of life. Almost all patients (95%) reported antimicrobial drug use. Of these 488 participants 280 (57%) reported the previous or current use of 5 or more categories of prostatitis related treatments. Conclusions: Chronic prostatitis/chronic pelvic pain syndrome is a multifactorial problem affecting men of all ages and demographics. Patients with the chronic pelvic pain syndrome have dismal quality of life and many have benefited only minimally from empirical, goal directed therapy. Long-term followup of this cohort may answer important questions on the natural treated history of this syndrome. C1 Northwestern Univ, Sch Med, Dept Urol, Chicago, IL 60611 USA. Univ Mississippi, Jackson, MS 39216 USA. Cleveland Clin Florida, Weston, ACT, Australia. Queens Univ, Dept Urol, Kingston, ON, Canada. Harvard Univ, Sch Med, Boston, MA USA. Univ Calif Los Angeles, Los Angeles, CA USA. NIDDK, Div Kidney Urol & Hematol Dis, NIH, Bethesda, MD USA. Univ Maryland, Baltimore, MD 21201 USA. Temple Univ, Philadelphia, PA 19122 USA. Univ Penn, Sch Med, Ctr Clin Epidemiol & Biostat, Philadelphia, PA 19104 USA. RP Schaeffer, AJ (reprint author), Northwestern Univ, Sch Med, Dept Urol, Tarry 11-715,303 E Chicago Ave, Chicago, IL 60611 USA. RI Landis, J. Richard/A-9330-2010 FU NIDDK NIH HHS [R01 DK53732, R01 DK53730, R01 DK53734, R01 DK53736, R01 DK53746, R01 DK53752] NR 20 TC 120 Z9 145 U1 0 U2 6 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-5347 J9 J UROLOGY JI J. Urol. PD AUG PY 2002 VL 168 IS 2 BP 593 EP 598 AR UNSP 0022-5347/02/1682-0593/0 DI 10.1016/S0022-5347(05)64686-1 PG 6 WC Urology & Nephrology SC Urology & Nephrology GA 573DD UT WOS:000176813300043 PM 12131316 ER PT J AU Dahle, SE Chokkalingam, AP Gao, YT Deng, J Stanczyk, FZ Hsing, AW AF Dahle, SE Chokkalingam, AP Gao, YT Deng, J Stanczyk, FZ Hsing, AW TI Body size and serum levels of insulin and leptin in relation to the risk of benign prostatic hyperplasia SO JOURNAL OF UROLOGY LA English DT Article DE prostate; prostatic hyperplasia; insulin; leptin; obesity ID HORMONE-BINDING GLOBULIN; FAT DISTRIBUTION; GROWTH-FACTORS; OBESITY; CANCER; RESISTANCE; SYSTEM; CHINA AB Purpose: Obesity has been implicated in the etiology of benign and malignant prostatic growth due to its influence on metabolic and endocrine changes. Because obesity is an important determinant of serum levels of insulin and leptin (the product of the obesity gene Ob), we investigated the role of obesity and serum levels of insulin and leptin in benign prostatic hyperplasia (BPH) etiology. Materials and Methods: Fasting serum levels of insulin and leptin as well as the body mass index, a measure of overall obesity, and waist-to-hip ratio, an indicator of abdominal obesity, were determined in 200 men newly diagnosed with BPH who were hospitalized for surgery and in 302 randomly selected healthy male subjects from the population in Shanghai, China. Results: A higher waist-to-hip ratio and higher serum insulin were significantly associated with an increased risk of BPH. Relative to men in the lowest waist-to-hip ratio quartile (less than 0.856) those in the highest quartile (greater than 0.923) were at 2.4-fold risk (odds ratio 2.42, 95% confidence interval [CI] 1.34 to 4.37, test for trend p = 0.01). Similarly relative to men in the lowest quartile of insulin (less than 5.87 muU. per ml.) those in the highest quartile (greater than 9.76 muU. per ml.) were at significantly increased risk (odds ratio 2.47, 95% CI 1.35 to 4.54, test for trend p = 0.009). The effect of insulin on BPH risk was more pronounced in men in low and middle tertiles of the waist-to-hip ratio (odds ratios comparing high to low insulin tertiles 2.8 and 2.7, respectively), while among men in the highest waist-to-hip ratio tertile insulin was not significantly associated with BPH risk. In contrast, we found no significant odds ratio comparing the highest to lowest quartiles of leptin (odds ratio 0.62, 95% CI 0.33 to 1.17) or body mass index (odds ratio 1.64, 95% CI 0.96 to 2.81). Conclusions: Our results suggest that abdominal obesity and increasing serum insulin, and possibly overall obesity but not serum leptin are associated with a higher risk of BPH. Further prospective and laboratory studies are needed to confirm these results and elucidate the underlying mechanisms. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ So Calif, Keck Sch Med, Dept Prevent Med, Los Angeles, CA USA. Univ So Calif, Keck Sch Med, Dept Obstet & Gynecol, Los Angeles, CA USA. Shanghai Canc Inst, Shanghai, Peoples R China. George Washington Univ, Sch Publ Hlth & Hlth Serv, Dept Epidemiol & Biostat, Washington, DC USA. RP Hsing, AW (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,Room 7058,MSC 7234, Bethesda, MD 20892 USA. NR 24 TC 131 Z9 150 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-5347 J9 J UROLOGY JI J. Urol. PD AUG PY 2002 VL 168 IS 2 BP 599 EP 604 AR UNSP 0022-5347/02/1682-0599/0 DI 10.1016/S0022-5347(05)64687-3 PG 6 WC Urology & Nephrology SC Urology & Nephrology GA 573DD UT WOS:000176813300044 PM 12131317 ER PT J AU Shamloul, AM Faggioli, F Keith, JM Hadidi, A AF Shamloul, AM Faggioli, F Keith, JM Hadidi, A TI A novel multiplex RT-PCR probe capture hybridization (RT-PCR-ELISA) for simultaneous detection of six viroids in four genera: Apscaviroid, Hostuviroid, Pelamoviroid, and Pospiviroid SO JOURNAL OF VIROLOGICAL METHODS LA English DT Article DE viroids; PSTVd; ASSVd; AFDVd; PBCVd; PLMVd; HSVd; detection; amplification; standard RT-PCR-ELISA; multiplex RT-PCR-ELISA; cDNA biotinylated probe ID POME FRUIT VIROIDS; POTATO SPINDLE TUBER; SENSITIVE DETECTION; NUCLEOTIDE-SEQUENCE; IDENTIFICATION; PEACH; DISEASE; STONE; PLUM AB A rapid and sensitive assay was developed for the detection and identification of viroids by standard or multiplex reverse transcription-polymerase chain reaction (RT-PCR)-probe capture hybridization (RT-PCR-ELISA). The assay was applied successfully for the detection and identification of the following six viroid species from infected tissues: Potato spindle tuber viroid (Pospiviroid), Peach latent mosaic viroid (Pelamoviroid), Apple scar skin viroid (Apscaviroid), Apple dimple fruit viroid (Apscaviroid), Pear blister canker viroid (Apscaviroid), and Hop stunt viroid (Hostuviroid). Total RNA was obtained from infected tissue by the Qiagen RNeasy kit and, then viroid cDNA was synthesized using viroid specific complementary DNA primer. To identify and differentiate the amplicons of the six viroids. each amplicon was digoxigenin (DIG)-labelled during the amplification process, and then detected by a colorimetric system using a biotinylated cDNA capture probe specific for each viroid. The results revealed that each capture probe hybridized only to its complementary DIG-labelled amplicon. Thus the six viroids can be detected and differentiated in a multiplex RT-PCR-ELISA assay, In the multiplex assay. cDNAs of six viroids were synthesized simultaneously in one tube, DIG-labelled during amplification, then a portion of the DIG-labelled amplified products was hybridized with selected capture probe, All the six viroid capture probes hybridized to their respective complementary DIG-labelled RT-PCR-amplified product. These findings are important for viroid detection and identification for studying host-viroid interactions and for management and control viroid diseases. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIDCR, Oral Infect & Immun Branch, Vaccine & Therapeut Dev Sect, NIH, Bethesda, MD 20892 USA. Ist Sperimentale Patol Vegetale, I-00156 Rome, Italy. Fraunhofer Ctr Mol Biotechnol, Newark, DE 19711 USA. USDA ARS, Fruit Lab, Beltsville, MD 20705 USA. RP Shamloul, AM (reprint author), S Valley Univ, Fac Sci, Dept Bot, Genet Lab, Sohag, Egypt. EM mshamloul@dir.nidcr.nih.gov NR 19 TC 26 Z9 31 U1 0 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-0934 J9 J VIROL METHODS JI J. Virol. Methods PD AUG PY 2002 VL 105 IS 1 BP 115 EP 121 AR PII S0166-0934(02)00090-3 DI 10.1016/S0166-0934(02)00090-3 PG 7 WC Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Virology GA 596EP UT WOS:000178151400012 PM 12176148 ER PT J AU Zhang, WH Hwang, CK Hu, WS Gorelick, RJ Pathak, VK AF Zhang, WH Hwang, CK Hu, WS Gorelick, RJ Pathak, VK TI Zinc finger domain of murine leukemia virus nucleocapsid protein enhances the rate of viral DNA synthesis in vivo SO JOURNAL OF VIROLOGY LA English DT Article ID HIV-1 REVERSE-TRANSCRIPTASE; PRIMER BINDING-SITE; RETROVIRAL SHUTTLE VECTOR; STRAND TRANSFER-REACTIONS; SINGLE REPLICATION CYCLE; ACID-CHAPERONE ACTIVITY; TYPE-1 GENOMIC RNA; BASIC-AMINO-ACIDS; STRONG-STOP DNA; IN-VITRO AB In vitro studies have indicated that retroviral nucleocapsid (NC) protein facilitates both DNA synthesis by reverse transcriptase (RT) and annealing of the nascent DNA with acceptor template. Increasing the rate of DNA synthesis is expected to reduce the frequency of RT template switching, whereas annealing the nascent DNA with acceptor template promotes template switching. We performed a mutational analysis of the murine leukemia virus (MLV) NC zinc finger domain to study its effect on RT template switching in vivo and to explore the role of NC during reverse transcription. The effects of NC mutations on RT template switching were determined by using a previously described in vivo direct-repeat deletion assay. A trans-complementation assay was also developed in which replication-defective NC mutants were rescued by coexpression of replication-defective RT mutants that provided wild-type NC in trans. We found that mutations in the MLV NC zinc finger domain increased the frequency of template switching approximately twofold. When a predicted stem-loop RNA secondary structure was introduced into the template RNA, the template-switching frequency increased 5-fold for wild-type NC and further increased up to an additional 6-fold for NC zinc finger domain mutants, resulting in an overall increase of as much as 30-fold. Thus, wild-type NC increased the efficiency with which RT was able to reverse transcribe through regions of RNA secondary structure that might serve as RT pause sites. These results provide the first in vivo evidence that NC enhances the rate of DNA synthesis by RT in regions of the template possessing stable RNA secondary structure. C1 NCI, HIV Drug Resistance Program, Frederick, MD 21702 USA. NCI, AIDS Vaccine Program, Sci Applicat Int Corp Frederick, Frederick, MD 21702 USA. W Virginia Univ, Dept Biol, Morgantown, WV 26506 USA. W Virginia Univ, Dept Microbiol & Immunol, Morgantown, WV 26506 USA. RP Pathak, VK (reprint author), NCI, HIV Drug Resistance Program, Bldg 535,Rm 334, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 65 TC 33 Z9 33 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7473 EP 7484 DI 10.1128/JVI.76.15.7473-7484.2002 PG 12 WC Virology SC Virology GA 571ZP UT WOS:000176749300012 PM 12097560 ER PT J AU Tomiyama, H Akari, H Adachi, A Takiguchi, M AF Tomiyama, H Akari, H Adachi, A Takiguchi, M TI Different effects of Nef-mediated HLA class I down-regulation on human immunodeficiency virus type I-specific CD8(+) T-cell cytolytic activity and cytokine production SO JOURNAL OF VIROLOGY LA English DT Article ID HIV-1 CTL EPITOPES; DENDRITIC CELLS; LYMPHOCYTES; MOLECULES; INFECTION; REPLICATION; PROGRESSION; AIDS; IDENTIFICATION; RESPONSES AB A previous study using a Nef-defective human immunodeficiency virus type I (HIV-1) mutant suggested that Nef-mediated down-regulation of HLA class I on the infected cell surface affects the cytolytic activity of HIV-1-specific cytotoxic T-lymphocyte (CTL) clones for HIV-1-infected primary CD4(+) T cells. We confirmed this effect by using a nef-mutant HIV-1 strain (NL-M20A) that expresses a Nef protein which does not induce down-regulation of HLA class I molecules but is otherwise functional. HIV-1-specific CTL clones were not able to kill primary CD4(+) T cells infected with a Nef-positive HIV-1 strain (NL-432) but efficiently lysed CD4(+) T cells infected with NL-M20A. Interestingly, CTL clones stimulated with NL-432-infected CD4(+) T cells were able to produce cytokines, albeit at a lower level than when stimulated with NL-M20A-infected CD4(+) T cells. This indicates that Nef-mediated HLA class I down-regulation affects CTL cytokine production to a lesser extent than cytolytic activity. Replication of NL-432 was partially suppressed in a coculture of HIV-1-infected CD4(+) T cells and HIV-1-specific CTL clones, while replication of NL-M20A was completely suppressed. These results suggest that HIV-1-specific CD8(+) T cells are able to partially suppress the replication of HIV-1 through production of soluble HIV-1-suppressive factors such as chemokines and gamma interferon. These findings may account for the mechanism whereby HIV-1-specific CD8(+) T cells are able to partially but not completely control HIV-1 replication in vivo. C1 Kumamoto Univ, Ctr AIDS Res, Div Viral Immunol, Kumamoto 8600811, Japan. Univ Tokushima, Sch Med, Dept Virol, Tokushima 7708503, Japan. NIAID, Mol Microbiol Lab, Viral Biochem Sect, NIH, Bethesda, MD 20892 USA. RP Takiguchi, M (reprint author), Kumamoto Univ, Ctr AIDS Res, Div Viral Immunol, 2-2-1 Honjo, Kumamoto 8600811, Japan. RI Takiguchi, Masafumi/E-7468-2013; Kumamoto University, CAIDS/G-8446-2013; Publications, Remarkable/I-1756-2013 NR 46 TC 64 Z9 66 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7535 EP 7543 DI 10.1128/JVI.76.15.7535-7543.2002 PG 9 WC Virology SC Virology GA 571ZP UT WOS:000176749300018 PM 12097566 ER PT J AU Rinke, CS Boyer, PL Sullivan, MD Hughes, SH Linial, ML AF Rinke, CS Boyer, PL Sullivan, MD Hughes, SH Linial, ML TI Mutation of the catalytic domain of the foamy virus reverse transcriptase leads to loss of processivity and infectivity SO JOURNAL OF VIROLOGY LA English DT Article ID DOUBLE-STRANDED DNA; HUMAN SPUMARETROVIRUS; ANGSTROM RESOLUTION; CRYSTAL-STRUCTURE; STERIC HINDRANCE; GENE-EXPRESSION; 3TC RESISTANCE; TYPE-1; REPLICATION; COMPLEX AB Foamy virus (FV) replication is resistant to most nucleoside analog reverse transcriptase (RT) inhibitors. In an attempt to create a 2',3'-dideoxy-3'-thiacytidine (3TC)-sensitive virus, the second residue in the highly conserved YXDD motif of simian foamy virus-chimpanzee (human isolate) [SFVcpz(hu)] RT was changed from Val (V) to Met (M). Unexpectedly, the resultant virus, SFVcpz(hu) RT-V313M, replicated poorly, and Met rapidly reverted to Val. Despite the presence of approximately 50% of wild-type RT activity in RT-V313M virions, full-length DNA products were not detected in transfected cells. Using purified recombinant enzymes, we found that the wild-type FV RT is significantly more processive than human immunodeficiency virus type 1 RT. However, the V313M mutant has about 40% of the wild-type level of FV RT activity and has a lower processivity than the wild-type FV enzyme. The V313M mutant RT is also relatively resistant to 3TC. These results suggest that the decrease in RT activity and processivity of FV RT-V313M prevents completion of reverse transcription and greatly diminishes viral replication. C1 Fred Hutchinson Canc Res Ctr, Div Basic Sci, Seattle, WA 98109 USA. Univ Washington, Dept Microbiol, Seattle, WA 98195 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Frederick, MD 21702 USA. RP Linial, ML (reprint author), Fred Hutchinson Canc Res Ctr, Div Basic Sci, 1100 Fairview Ave N, Seattle, WA 98109 USA. FU NCI NIH HHS [R01 CA018282, CA-18282, CA-09229, T32 CA009229] NR 31 TC 27 Z9 27 U1 1 U2 4 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7560 EP 7570 DI 10.1128/JVI.76.15.7560-7570.2002 PG 11 WC Virology SC Virology GA 571ZP UT WOS:000176749300021 PM 12097569 ER PT J AU Amara, RR Villinger, F Staprans, SI Altman, JD Montefiori, DC Kozyr, NL Xu, Y Wyatt, LS Earl, PL Herndon, JG McClure, HM Moss, B Robinson, HL AF Amara, RR Villinger, F Staprans, SI Altman, JD Montefiori, DC Kozyr, NL Xu, Y Wyatt, LS Earl, PL Herndon, JG McClure, HM Moss, B Robinson, HL TI Different patterns of immune responses but similar control of a simian-human immunodeficiency virus 89.6P mucosal challenge by modified vaccinia virus Ankara (MVA) and DNA/MVA vaccines SO JOURNAL OF VIROLOGY LA English DT Article ID POLYMERASE CHAIN-REACTION; RHESUS-MONKEYS; T-CELLS; NEUTRALIZING ANTIBODIES; MACAQUES; AIDS; VACCINATION; SIV; REPLICATION; PREVENTION AB Recently we demonstrated the control of a mucosal challenge with a pathogenic chimera of simian and human immunodeficiency virus (SHIV-89.6P) by priming with a Gag-Pol-Env-expressing DNA and boosting with a Gag-Pol-Env-expressing recombinant modified vaccinia virus Ankara (DNA/MVA) vaccine. Here we evaluate the ability of the MVA component of this vaccine to serve as both a prime and a boost for an AIDS vaccine. The same immunization schedule, MVA dose, and challenge conditions were used as in the prior DNA/MVA vaccine trial. Compared to the DNA/MVA vaccine, the MVA-only vaccine raised less than 1/10 the number of vaccine-specific T cells but 10-fold-higher titers of binding antibody for Env. Postchallenge, the animals vaccinated with WA alone increased their CD8 cell numbers to levels that were similar to those seen in DNA/MVA-vaccinated animals. However, they underwent a slower emergence and contraction of antiviral CD8 T cells and were slower to generate neutralizing antibodies than the DNA/MVA-vaccinated animals. Despite this, by 5 weeks postchallenge, the MVA-only-vaccinated animals had achieved as good control of the viral infection as the DNA/MVA group, a situation that has held up to the present time in the trial (48 weeks postchallenge). Thus, MVA vaccines, as well as DNA/MVA vaccines, merit further evaluation for their ability to control the current AIDS pandemic. C1 Emory Univ, Vaccine Res Ctr, Atlanta, GA 30329 USA. Emory Univ, Yerkes Reg Primate Res Ctr, Atlanta, GA 30329 USA. Emory Univ, Sch Med, Dept Microbiol & Immunol, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Pathol & Lab Med, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA. Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA. NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Robinson, HL (reprint author), 954 Gatewood Rd NE, Atlanta, GA 30329 USA. FU NCRR NIH HHS [P51 RR000165, P51 RR00165]; NIAID NIH HHS [AI 85343, P01 AI 43045, P01 AI043045]; NIDA NIH HHS [P30 DA 12121] NR 25 TC 115 Z9 119 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7625 EP 7631 DI 10.1128/JVI.76.15.7625-7631.2002 PG 7 WC Virology SC Virology GA 571ZP UT WOS:000176749300028 PM 12097576 ER PT J AU Yang, GS Schmidt, M Yan, ZY Lindbloom, JD Harding, TC Donahue, BA Engelhardt, JF Kotin, R Davidson, BL AF Yang, GS Schmidt, M Yan, ZY Lindbloom, JD Harding, TC Donahue, BA Engelhardt, JF Kotin, R Davidson, BL TI Virus-mediated transduction of murine retina with adeno-associated virus: Effects of viral capsid and genome size SO JOURNAL OF VIROLOGY LA English DT Article ID RECOMBINANT ADENOASSOCIATED VIRUS; GENE-TRANSFER; EFFICIENT TRANSDUCTION; AIRWAY EPITHELIA; AAV SEROTYPES; MOUSE RETINA; IN-VIVO; VECTORS; TYPE-5; EXPRESSION AB Gene therapy vectors based on adeno-associated viruses (AAVs) show promise for the treatment of retinal degenerative diseases. In prior work, subretinal injections of AAV2, AAV5, and AAV2 pseudotyped with AAV5 capsids (AAV2/5) showed variable retinal pigmented epithelium (RPE) and photoreceptor cell transduction, while AAV2/1 predominantly transduced the RPE. To more thoroughly compare the efficiencies of gene transfer of AAV2, AAV3, AAV5, and AAV6, we quantified, using stereological methods, the kinetics and efficiency of AAV transduction to mouse photoreceptor cells. We observed persistent photoreceptor and RPE transduction by AAV5 and AAV2 up to 31 weeks and found that AAV5 transduced a greater volume than AAV2. AAV5 containing full-length or half-length genomes and AAV2/5 transduced comparable numbers of photoreceptor cells with similar rates of onset of expression. Compared to AAV2, AAV5 transduced significantly greater numbers of photoreceptor cells at 5 and 15 weeks after surgery (greater than 1,000 times and up to 400 times more, respectively). Also, there were 30 times more genome copies in eyes injected with AAV2/5 than in eyes injected with AAV2. Comparing AAVs with half-length genomes, AAV5 transduced only four times more photoreceptor cells than AAV2 at 5 weeks and nearly equivalent numbers at 15 weeks. The enhancement of transduction was seen at the DNA level, with 50 times more viral genome copies in retinas injected with AAV having short genomes than in retinas injected with AAV containing full-length ones. Subretinal injection of AAV2/6 showed only RPE transduction at 5 and 15 weeks, while AAV2/3 did not transduce retinal cells. We conclude that varying genome length and AAV capsids may allow for improved expression and/or gene transfer to specific cell types in the retina. C1 Univ Iowa, Coll Med, Dept Internal Med, Program Gene Therapy, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Otolaryngol, Program Gene Therapy, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Neurol, Program Gene Therapy, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Physiol & Biophys, Program Gene Therapy, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Anat & Cell Biol, Program Gene Therapy, Iowa City, IA 52242 USA. NHLBI, Bethesda, MD 20892 USA. Cell Genesys Inc, Foster City, CA USA. RP Davidson, BL (reprint author), Univ Iowa, Coll Med, Dept Internal Med, Program Gene Therapy, 200 EMRB, Iowa City, IA 52242 USA. RI kotin, robert/B-8954-2008 FU NHLBI NIH HHS [R01 HL058340, HL58340]; NICHD NIH HHS [HD33531, R01 HD033531]; NIDCD NIH HHS [DC00040, T32 DC000040]; NIDDK NIH HHS [DK54759, P30 DK054759] NR 40 TC 114 Z9 118 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7651 EP 7660 DI 10.1128/JVI.76.15.7651-7660.2002 PG 10 WC Virology SC Virology GA 571ZP UT WOS:000176749300031 PM 12097579 ER PT J AU Farrell, KB Russ, JL Murthy, RK Eiden, MV AF Farrell, KB Russ, JL Murthy, RK Eiden, MV TI Reassessing the role of region A in Pit1-mediated viral entry SO JOURNAL OF VIROLOGY LA English DT Article ID APE LEUKEMIA-VIRUS; PROTEIN SECONDARY STRUCTURE; CELL-SURFACE RECEPTORS; SUBGROUP-B; PHOSPHATE TRANSPORTER; TRANSMEMBRANE TOPOLOGY; ENDOPLASMIC-RETICULUM; RETROVIRUS RECEPTOR; MEMBRANE-PROTEINS; INFECTION AB The mammalian gammaretroviruses gibbon ape leukemia virus (GALV) and feline leukemia virus subgroup B (FeLV-B) can use the same receptor, Pit1, to infect human cells. A highly polymorphic nine-residue sequence within Pit1, designated region A, has been proposed as the virus binding site, because mutations in this region abolish Pit1-mediated cellular infection by GALV and FeLV-B. However, a direct correlation between region A mutations deleterious for infection and loss of virus binding has not been established. We report that cells expressing a Pit1 protein harboring mutations in region A that abolish receptor function retain the ability to bind virus, indicating that Pit1 region A is not the virus binding site. Furthermore, we have now identified a second region in Pit1, comprising residues 232 to 260 (region B), that is required for both viral entry and virus binding. Epitope-tagged Pit1 proteins were used to demonstrate that mutations in region B result in improper orientation of Pit1 in the cell membrane. Compensatory mutations in region A can restore proper orientation and full receptor function to these region B mutants. Based on these results, we propose that region A of Pit1 confers competence for viral entry by influencing the topology of the authentic binding site in the membrane and hence its accessibility to a viral envelope protein. Based on glycosylation studies and results obtained by using N- and C-terminal epitope-tagged Pit1, region A and region B mutants, and the transmembrane helices predicted with the PHD PredictProtein algorithm, we propose a new Pit1 topology model. C1 NIMH, Unit Mol Virol, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. RP Eiden, MV (reprint author), NIMH, Unit Mol Virol, Lab Cellular & Mol Regulat, Bldg 36,Room 2A11,36 Convent Dr,MSC 4068, Bethesda, MD 20892 USA. NR 38 TC 28 Z9 30 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7683 EP 7693 DI 10.1128/JVI.76.15.7683-7693.2002 PG 11 WC Virology SC Virology GA 571ZP UT WOS:000176749300034 PM 12097582 ER PT J AU Husain, M Moss, B AF Husain, M Moss, B TI Similarities in the induction of post-Golgi vesicles by the vaccinia virus F13L protein and phospholipase D SO JOURNAL OF VIROLOGY LA English DT Article ID EXTRACELLULAR ENVELOPED VIRUS; ACTIN-CONTAINING MICROVILLI; TO-CELL SPREAD; INTRACELLULAR MOVEMENT; CATALYTIC MOTIF; D DEFINES; GLYCOPROTEIN; MEMBRANE; GENE; LOCALIZATION AB Intracellular mature vaccinia virions are wrapped by cisternae, derived from virus-modified trans-Golgi or endosomal membranes, and then transported via microtubules to the cell periphery. Two viral proteins, encoded by the F13L and B5R open reading frames, are essential for the membrane-wrapping step. Previous transfection studies indicated that F13L induces the formation of post-Golgi vesicles that incorporate the B5R protein and that this activity depends on an intact F13L phospholipase motif. Here we show that the F13L protein has a general effect on the trafficking of integral membrane proteins from the Golgi apparatus, as both the vaccinia virus A36R protein and the vesicular stomatitis virus G protein also colocalized with the F13L protein in vesicles. In addition, increased expression of cellular phospholipase D, which has a similar phospholipase motif as, but little amino acid sequence identity with, F13L, induced post-Golgi vesicles that contained B5R and A36R proteins. Butanol-1, which prevents the formation of phosphatidic acid by phospholipase D and specifically inhibits phospholipase D-mediated vesicle formation, also inhibited F13L-induced vesicle formation, whereas secondary and tertiary alcohols had no effect. Moreover, inhibition of phospholipase activity by butanol-1 also reduced plaque size and decreased the formation of extracellular vaccinia virus without affecting the yield of intracellular mature virus. Phospholipase D, however, could not complement a vaccinia virus F13L deletion mutant, indicating that F13L has additional virus-specific properties. Taken together, these data support an important role for F13L in inducing the formation of vesicle precursors of the vaccinia virus membrane via phospholipase activity or activation. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, 4 Ctr Dr,MSC 0445, Bethesda, MD 20892 USA. NR 53 TC 33 Z9 34 U1 1 U2 5 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7777 EP 7789 DI 10.1128/JVI.76.15.7777-7789.2002 PG 13 WC Virology SC Virology GA 571ZP UT WOS:000176749300042 PM 12097590 ER PT J AU Cheng, N Trus, BL Belnap, DM Newcomb, WW Brown, JC Steven, AC AF Cheng, N Trus, BL Belnap, DM Newcomb, WW Brown, JC Steven, AC TI Handedness of the herpes simplex virus capsid and procapsid SO JOURNAL OF VIROLOGY LA English DT Article ID CRYOELECTRON MICROSCOPY; BIOLOGICAL MACROMOLECULES; ELECTRON CRYOMICROSCOPY; 3-DIMENSIONAL STRUCTURE; CYTOMEGALOVIRUS; MATURATION; VISUALIZATION; PENTONS; TYPE-1; DNA AB The capsid of herpes simplex virus has an icosahedral surface lattice with a nonskew triangulation number, T=16. Nevertheless, the proteins arrayed on this lattice necessarily have an intrinsic handedness. We have determined the handedness of both the herpes simplex virus type 1 capsid and its precursor procapsid by a cryoelectron microscopic tilting method. C1 NIAMSD, Struct Biol Lab, NIH, Bethesda, MD 20892 USA. NIAMSD, Computat Biosci & Engn Lab, Ctr Informat Technol, NIH, Bethesda, MD 20892 USA. Univ Virginia, Dept Microbiol, Charlottesville, VA 22908 USA. RP Steven, AC (reprint author), NIAMSD, Struct Biol Lab, NIH, Bldg 50,Rm 1517,MSC 8025, Bethesda, MD 20892 USA. FU NIAID NIH HHS [R01 AI041644, R-01 AI41644-04] NR 28 TC 33 Z9 33 U1 0 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7855 EP 7859 DI 10.1128/JVI.76.15.7855-7859.2002 PG 5 WC Virology SC Virology GA 571ZP UT WOS:000176749300049 PM 12097597 ER PT J AU Center, RJ Leapman, RD Lebowitz, J Arthur, LO Earl, PL Moss, B AF Center, RJ Leapman, RD Lebowitz, J Arthur, LO Earl, PL Moss, B TI Oligomeric structure of the human immunodeficiency virus type I envelope protein on the virion surface SO JOURNAL OF VIROLOGY LA English DT Article ID MEMBRANE-FUSION; CRYSTAL-STRUCTURE; GLYCOPROTEIN ECTODOMAIN; MONOCLONAL-ANTIBODIES; DISULFIDE BONDS; GP41 ECTODOMAIN; RESOLUTION; BINDING; GP160; GP120 AB The envelope protein (Env) of human immunodeficiency virus type 1 forms homo-oligomers in the endoplasmic reticullum. The oligomeric structure of Env is maintained after cleavage in a Golgi compartment and transport to the surfaces of infected cells, where incorporation into budding virions takes place. Here, we use biophysical techniques to assess the oligomeric valency of virion-associated Env prior to fusion activation. Virion-associated Env oligomers were stabilized by chemical cross-linking prior to detergent extraction and were purified by immunoaffinity chromatography. Gel filtration revealed a single predominant oligomeric species, and sedimentation equilibrium analysis-derived mass values indicated a trimeric structure. Determination of the masses of individual Env molecules by scanning transmission electron microscopy demonstrated that virion-associated Env was trimeric, and a triangular morphology was observed in 20 to 30% of the molecules. These results, which firmly establish the oligomeric structure of human immunodeficiency virus virion-associated Env, parallel those of our previous analysis of the simian immunodeficiency virus Env. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. NIH, Div Bioengn & Phys Sci, Off Res Serv, Bethesda, MD 20892 USA. NCI, AIDS Vaccine Program, Sci Applicat Int Corp, Frederick, MD 21702 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, Bldg 4,Room 229,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 37 TC 83 Z9 85 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 15 BP 7863 EP 7867 DI 10.1128/JVI.76.15.7863-7867.2002 PG 5 WC Virology SC Virology GA 571ZP UT WOS:000176749300051 PM 12097599 ER PT J AU Peterson, KE Stromnes, I Messer, R Hasenkrug, K Chesebro, B AF Peterson, KE Stromnes, I Messer, R Hasenkrug, K Chesebro, B TI Novel role of CD8(+) T cells and major histocompatibility complex class I genes in the generation of protective CD4(+) Th1 responses during retrovirus infection in mice SO JOURNAL OF VIROLOGY LA English DT Article ID FRIEND-VIRUS MODEL; GAMMA-INTERFERON; INDUCED LEUKEMIA; SPONTANEOUS-RECOVERY; MURINE RETROVIRUS; SUBSET ACTIVATION; LEISHMANIA-MAJOR; IMMUNE-RESPONSE; DENDRITIC CELLS; MHC CONTROL AB CD4(+) Th1 responses to virus infections are often necessary for the development and maintenance of virus-specific CD8(+) T-cell responses. However, in the present study with Friend murine retrovirus (FV), the reverse was also found to be true. In the absence of a responder H-2(b) allele at major histocompatibillity complex (MHC) class 11 loci, a single H-2D(b) MHC class I allele was sufficient for the development of a CD4(+) Th1 response to FV. This effect of H-2D(b) on CD4(+) T-cell responses was dependent on CD8(+) T cells, as demonstrated by depletion studies. A direct effect of CD8(+) T-cell help in the development of CD4(+) Th1 responses to FV was also shown in vaccine studies. Vaccination of nonresponder H-2(a/a) mice induced FV-specific responses of H-2D(d)-restricted CD8(+) cytotoxic T lymphocytes (CTL). Adoptive transfer of vaccine-primed CD8(+) T cells to naive H-2(a/a) mice prior to infection resulted in the generation of FV-specific CD4(+) Th1 responses. This novel helper effect of CD8(+) T cells could be an important mechanism in the development of CD4(+) Th1 responses following vaccinations that induce CD8(+) CTL responses. The ability of MHC class I genes to facilitate CD4(+) Th1 development could also be considerable evolutionary advantage by allowing a wider variety of MHC genotypes to generate protective immune responses against intracellular pathogens. C1 NIAID, Rocky Mt Labs, Lab Persistent Viral Dis, NIH, Hamilton, MT 59840 USA. RP Chesebro, B (reprint author), NIAID, Rocky Mt Labs, Lab Persistent Viral Dis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. RI Peterson, Karin/D-1492-2016 OI Peterson, Karin/0000-0003-4177-7249 NR 46 TC 17 Z9 18 U1 0 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 16 BP 7942 EP 7948 DI 10.1128/JVI.76.16.7942-7948.2002 PG 7 WC Virology SC Virology GA 577EX UT WOS:000177049500003 PM 12133998 ER PT J AU Lauring, AS Cheng, HH Eiden, MV Overbaugh, J AF Lauring, AS Cheng, HH Eiden, MV Overbaugh, J TI Genetic and biochemical analyses of receptor and cofactor determinants for T-cell-tropic feline leukemia virus infection SO JOURNAL OF VIROLOGY LA English DT Article ID ECOTROPIC MURINE RETROVIRUSES; SUBGROUP-B; ENVELOPE PROTEIN; BINDING DOMAIN; HOST-RANGE; PHOSPHATE TRANSPORTERS; SURFACE GLYCOPROTEIN; MEMBRANE-FUSION; HAMSTER-CELLS; ENTRY AB Entry by retroviruses is mediated through interactions between the viral envelope glycoprotein and the host cell receptor(s). We recently identified two host cell proteins, FeLIX and Pit1, that are necessary for infection by cytopathic, T-cell-tropic feline leukemia viruses (FeLV-T). Pit1 is a classic multiple transmembrane protein used as a receptor by several other simple retroviruses, including subgroup B FeLV (FeLV-B), and FeLIX is a secreted cellular protein expressed from endogenous FeLV-related sequences (enFeLV). FeLIX is nearly identical to FeLV-B envelope sequences that encode the N-terminal half of the viral surface unit (SU), because these FeLV-B sequences are acquired by recombination with enFeLV. FeLV-B SUs can functionally substitute for FeLIX in mediating FeLV-T infection. Both of these enFeLV-derived cofactors can efficiently facilitate FeLV-T infection only of cells expressing Pit1, not of cells expressing the related transport protein Pit2. We therefore have used chimeric Pit1/Pit2 receptors to map the determinants for cofactor binding and FeLV-T infection. Three distinct determinants appear to be required for cofactor-dependent infection by FeLV-T. We also found that Pit1 sequences within these same domains were required for binding by FeLIX to the Pit receptor. In contrast, these determinants were not all required for receptor binding by the FeLV-B SU cofactors used in this study. These data indicate that cofactor binding is not sufficient for FeLV-T infection and suggest that there may be a direct interaction between FeLV-T and the Pit1 receptor. C1 Fred Hutchinson Canc Res Ctr, Div Human Biol, Seattle, WA 98109 USA. Univ Washington, Program Mol & Cellular Biol, Seattle, WA 98195 USA. Natl Inst Mental Hlth, Lab Cellular & Mol Regulat, Bethesda, MD USA. RP Overbaugh, J (reprint author), Fred Hutchinson Canc Res Ctr, Div Human Biol, 1100 Fairview Ave,N,C3-168, Seattle, WA 98109 USA. FU NCI NIH HHS [CA 51080, T32 CA009229, R01 CA051080, 2 T32 CA09229] NR 60 TC 11 Z9 11 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 16 BP 8069 EP 8078 DI 10.1128/JVI.76.16.8069-8078.2002 PG 10 WC Virology SC Virology GA 577EX UT WOS:000177049500017 PM 12134012 ER PT J AU Jung, YT Lyu, MS Buckler-White, A Kozak, CA AF Jung, YT Lyu, MS Buckler-White, A Kozak, CA TI Characterization of a polytropic murine leukemia virus proviral sequence associated with the virus resistance gene Rmcf of DBA/2 mice SO JOURNAL OF VIROLOGY LA English DT Article ID CELL-SURFACE RECEPTOR; MUS CASTANEUS; MOUSE CELLS; INFECTION; SUSCEPTIBILITY; CLONING; FV-4; ERYTHROLEUKEMIA; IDENTIFICATION; GLYCOPROTEIN AB The DBA/2 mouse Rmcf gene is responsible for in vivo and in vitro resistance to infection by the polytropic mink cell focus-forming (MCF) virus subgroup of murine leukemia viruses (MLVs). Previous studies suggested that Rmcf resistance is mediated by expression of an interfering MCF MLV envelope (Env) gene. To characterize this env gene, we examined resistance in crosses between Rmcf(r) DBA/2 mice and Mus castaneus, a species that lacks endogenous MCF env sequences. In backcross progeny, inheritance of Rmcf resistance correlated with inheritance of a specific endogenous MCF virus env-containing 4.6-kb EcoRI fragment. This fragment was present in the DBA/2N substrain with Rmcf-mediated resistance but not in virus-susceptible DBA/2J substrain mice. This fragment contains a provirus with a 5' long terminal repeat and the 5' half of env; the gag and pol genes have been partially deleted. The Env sequence is identical to that of a highly immunogenic viral glycoprotein expressed in the DBA/2 cell line L5178Y and closely resembles the env genes of modified polytropic proviruses. The coding sequence for the full-length Rmcf Env surface subunit was amplified from DNAs from virus-resistant backcross mice and was cloned into an expression vector. NIH 3T3 and BALB 3T3 cells stably transfected with this construct showed significant resistance to infection by MCF MLV but not by amphotropic MLV. This study identifies an Rmcf-linked MCF provirus and indicates that, like the ecotropic virus resistance gene Fv4, Rmcf may mediate resistance through an interference mechanism. C1 NIAID, LMM, NIH, Bethesda, MD 20892 USA. RP Kozak, CA (reprint author), NIAID, LMM, NIH, Bldg 4,Room 329,4 Ctr Dr,MSC 0460, Bethesda, MD 20892 USA. NR 33 TC 22 Z9 22 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 16 BP 8218 EP 8224 DI 10.1128/JVI.76.16.8218-8224.2002 PG 7 WC Virology SC Virology GA 577EX UT WOS:000177049500032 PM 12134027 ER PT J AU Mastakov, MY Baer, K Symes, CW Leichtlein, CB Kotin, RM During, MJ AF Mastakov, MY Baer, K Symes, CW Leichtlein, CB Kotin, RM During, MJ TI Immunological aspects of recombinant adeno-associated virus delivery to the mammalian brain SO JOURNAL OF VIROLOGY LA English DT Article ID MEDIATED GENE-TRANSFER; ADENOASSOCIATED VIRAL VECTORS; IN-VIVO EXPRESSION; FACTOR-IX; TRANSGENE EXPRESSION; TRANSIENT IMMUNOSUPPRESSION; NEUTRALIZING ANTIBODIES; IMMUNE-RESPONSES; RABBIT AIRWAY; AAV VECTOR AB Recombinant adeno-associated viruses (rAAV) are highly efficient vectors for gene delivery into the central nervous system (CNS). However, host inflammatory and immune responses may play a critical role in limiting the use of rAAV vectors for gene therapy and functional genomic studies in vivo. Here, we evaluated the effect of repeated injections of five rAAV vectors expressing different genetic sequences (coding or noncoding) in a range of combinations into the rat brain. Specifically, we wished to determine whether a specific immune or inflammatory response appeared in response to the vector and/or the transgene protein after repeated injections under conditions of mannitol coinjection. We show that readministration of the same rAAV to the CNS is possible if the interval between the first and second injection is more than 4 weeks. Furthermore, our data demonstrate that rAAV vectors carrying different genetic sequences can be administered at intervals of 2 weeks. Our data therefore suggest that the AAV capsid structure is altered by the vector genetic sequence, such that secondary structures of the single-stranded genome have an impact on the antigenicity of the virus. This study provides guidelines for more rational design of gene transfer studies in the rodent brain and, in addition, suggests the use of repeated administration of rAAV as a viable form of therapy for the treatment of chronic diseases. C1 Jefferson Med Coll, Dept Neurosurg, CNS Gene Therapy Ctr, Philadelphia, PA 19107 USA. Univ Auckland, Fac Med & Hlth Sci, Div Mol Med, Funct Genom & Translat Neurosci Lab, Auckland 1, New Zealand. NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. RP During, MJ (reprint author), Jefferson Med Coll, Dept Neurosurg, CNS Gene Therapy Ctr, Philadelphia, PA 19107 USA. RI kotin, robert/B-8954-2008 NR 54 TC 76 Z9 80 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD AUG PY 2002 VL 76 IS 16 BP 8446 EP 8454 DI 10.1128/JVI.76.16.8446-8454.2002 PG 9 WC Virology SC Virology GA 577EX UT WOS:000177049500052 PM 12134047 ER PT J AU Karamessinis, PM Tzinia, AK Kitsiou, PV Stetler-Stevenson, WG Michael, AF Fan, WW Zhou, B Margaritis, LH Tsilibary, EC AF Karamessinis, PM Tzinia, AK Kitsiou, PV Stetler-Stevenson, WG Michael, AF Fan, WW Zhou, B Margaritis, LH Tsilibary, EC TI Proximal tubular epithelial cell integrins respond to high glucose by altered cell-matrix interactions and differentially regulate matrixin expression SO LABORATORY INVESTIGATION LA English DT Article ID DEPENDENT DIABETES-MELLITUS; COLLAGENASE GENE-EXPRESSION; HUMAN MESANGIAL CELLS; IV COLLAGEN; BASEMENT-MEMBRANE; EXTRACELLULAR-MATRIX; SIGNAL-TRANSDUCTION; ALPHA-3-BETA-1 INTEGRIN; FIBRONECTIN; ADHESION AB Thickening of the tubular basement membrane (TBM) occurs in diabetic nephropathy, but the effects of high glucose on the functional aspects of proximal tubular epithelial cells are not clearly understood. In the present study, we examined the effects of elevated glucose concentrations on (a) integrin expression by human proximal tubular epithelial cells (HK-2) and integrin-mediated interactions with type IV collagen (coIIV) and laminin, major components of TBM; (b) the expression of matrixins/matrix metalloproteinases (MMPs), which is regulated by integrins; and (c) the expression of tissue inhibitors of metalloproteinases (TIMPs). HK-2 cells cultured in 25 mM glucose underwent a reduction of the expression of alpha(3), beta1, alphavbeta3, and alpha5 integrin subunits, with a concomitant increase of the alpha2 subunit, compared with cells grown in 5 mM glucose. Adhesion experiments demonstrated that high glucose led to increased cell adhesion on either coIIV or laminin. Experiments of competition of adhesion using anti-integrin antibodies indicated that HK-2 cells in 5 mM glucose used mainly alphavbeta3 and alpha5beta1 integrins to adhere to coIIV, whereas in 25 mM glucose they additionally used alpha2beta1. In the case of laminin, a beta1-mediated adhesion was observed when HIK-2 cells were in 5 mM glucose, whereas in 25 mm glucose, alpha2beta1 and alphavbeta3 were also involved. Elevated glucose concentrations resulted in decreased expression of MMP-9 and MMP-2, whereas an increase in TIMP-1 and a decrease in TIMP-2 expression were observed. We also examined which integrins mediated the expression and secretion of matrixins MMP-2 and MMP-9. Ligation of alpha3beta1 with mAbs resulted in induction of MMP-2 expression and secretion, whereas antibody ligation of alphavbeta3 led to down-regulation of MMP-9. The above data implicate integrins of proximal tubular epithelial cells in the regulation of MMPs and in the development of TBM thickening in diabetic nephropathy. C1 Univ Athens, Inst Biol, Natl Ctr Sci Res Demokritos, GR-15310 Athens, Greece. Univ Athens, Fac Biol, Dept Cell Biol & Biophys, GR-15310 Athens, Greece. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Univ Minnesota, Sch Med, Dept Pediat, Minneapolis, MN 55455 USA. RP Tsilibary, EC (reprint author), Univ Athens, Inst Biol, Natl Ctr Sci Res Demokritos, GR-15310 Athens, Greece. RI Stetler-Stevenson, William/H-6956-2012 OI Stetler-Stevenson, William/0000-0002-5500-5808 FU NIAID NIH HHS [AI0708] NR 73 TC 22 Z9 29 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0023-6837 J9 LAB INVEST JI Lab. Invest. PD AUG PY 2002 VL 82 IS 8 BP 1081 EP 1093 DI 10.1097/01.LAB.0000022224.86237.31 PG 13 WC Medicine, Research & Experimental; Pathology SC Research & Experimental Medicine; Pathology GA 586DY UT WOS:000177570400013 PM 12177247 ER PT J AU Crump, M Couban, S Meyer, R Rudinskas, L Zanke, B Gluck, S Maksymiuk, A Hoskins, P Matthews, S Eisenhauer, E AF Crump, M Couban, S Meyer, R Rudinskas, L Zanke, B Gluck, S Maksymiuk, A Hoskins, P Matthews, S Eisenhauer, E TI Phase II study of sequential topotecan and etoposide in patients with intermediate grade non-Hodgkin's lymphoma: A National Cancer Institute of Canada Clinical Trials Group study SO LEUKEMIA & LYMPHOMA LA English DT Article DE topoisomerase I; relapsed; non-Hodgkin's lymphoma; topotecan; etoposide ID BONE-MARROW TRANSPLANTATION; CELL LUNG-CANCER; PROSPECTIVE RANDOMIZED TRIAL; COOPERATIVE STUDY-GROUP; STANDARD REGIMEN CHOP; DNA TOPOISOMERASE-II; ORAL ETOPOSIDE; SALVAGE CHEMOTHERAPY; LEUKEMIA; CAMPTOTHECIN AB Preliminary results indicate that inhibitors of the nuclear enzyme topoisomerase (topo) I, such as topotecan, may be active in non-Hodgkin's lymphoma (NHL). Pre-clinical studies have shown sequential administration of a topo I and II inhibitor has supra-additive anti-tumor effects in some model systems, and that greater cytotoxicity occurs if the topo I inhibitor is given first. We enrolled 22 eligible patients with relapsed or refractory intermediate grade NHL in a phase II study of sequential administration of topotecan 1.25 mg/m(2) days 1-5 and etoposide 50 mg po b.i.d. days 6-12, every 28 days without G-CSF. Most patients had diffuse large B-cell lymphoma and all had received only one prior regimen (CHOP, 20 patients, or equivalent, 2 patients). Patients with stable or responding disease were allowed to proceed to high-dose therapy and autologous stem-cell transplant after 2 cycles of therapy. The 22 patients received a total of 62 cycles of topotecan + etoposide (median 2, range 1-6), and 4/22 completed all six planned cycles. Hematologic toxicity was significant and resulted in incomplete etoposide dosing in half of all cycles in 16/22 patients. Nineteen of twenty-two patients had grade 3/4 neutropenia, 12 had grade 3/4 thrombocytopenia, and 6 grade 3/4 anemia. Eleven patients had at least one episode of febrile neutropenia or had documented infection. Non-hematologic toxicity was mild. Four patients had a partial response (PR) (18.2%), nine had stable disease and seven progressed; three patients with stable disease went on to ABMT. The combination of topotecan and etoposide as given in this study has modest activity in relapsed/refractory aggressive histology NHL, and produces marked myelosuppression. Other doses and schedules combining topo I and II inhibitors, or topo I inhibitors with alkylating agents, should be explored with the addition of hematopoietic growth factors in this patient population. C1 Princess Margaret Hosp, Dept Med Oncol & Hematol, Toronto, ON M5G 2M9, Canada. Queen Elizabeth II Med Ctr, Dept Hematol, Halifax, NS, Canada. Hamilton Hlth Sci Corp, Hamilton, ON, Canada. Humber River Reg Hosp, Dept Med Oncol, Toronto, ON, Canada. Tom Baker Canc Clin, Dept Med, Calgary, AB, Canada. Saskatoon Canc Ctr, Sect Med Oncol, Saskatoon, SK, Canada. BC Canc Agcy, Dept Med Oncol, Vancouver, BC, Canada. NCI, Clin Trials Grp, Kingston, ON, Canada. RP Crump, M (reprint author), Princess Margaret Hosp, Dept Med Oncol & Hematol, 610 Univ Ave,Room 5-108, Toronto, ON M5G 2M9, Canada. NR 37 TC 15 Z9 17 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI ABINGDON PA 4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLAND SN 1042-8194 J9 LEUKEMIA LYMPHOMA JI Leuk. Lymphoma PD AUG PY 2002 VL 43 IS 8 BP 1581 EP 1587 DI 10.1080/1042819021000002901 PG 7 WC Oncology; Hematology SC Oncology; Hematology GA 580FF UT WOS:000177223900008 PM 12400600 ER PT J AU Hori, M Qi, CF Torrey, TA Huppi, K Morse, HC AF Hori, M Qi, CF Torrey, TA Huppi, K Morse, HC TI The Bcl6 locus is not mutated in mouse B-cell lineage lymphomas SO LEUKEMIA RESEARCH LA English DT Article DE Bcl6; Burkitt lymphoma; diffuse large cell lymphoma; plasmacytoma ID NON-HODGKINS-LYMPHOMAS; SOMATIC HYPERMUTATION; 5'-NONCODING REGION; BURKITTS-LYMPHOMA; POINT MUTATIONS; GENE-EXPRESSION; LEUKEMIA; MICE; TRANSLOCATIONS; REARRANGEMENTS AB In normal human germinal centre (GC) B-cells and post-GC B-cell lymphomas, a region in the first intron of the BCL6 gene, termed the major mutations cluster (MMC) exhibits somatic point mutations and deletions with patterns very similar to those seen in the variable regions of immunoglobulin (Ig) genes. In studies of mouse post-GC diffuse large cell lymphoma, Burkitt lymphomas, and plasmacytomas, direct sequencing or cold SSCP analyses revealed no mutations within a 686-bp region in Bc16 intron 1 with 72% identity to the human MMC. The mouse Bc16 locus must be inaccessible to the mutational machinery responsible for somatic mutations of Ig and BCL6 in humans. Published by Elsevier Science Ltd. C1 NIAID, Immunopathol Lab, Bethesda, MD 20892 USA. NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. RP Hori, M (reprint author), NIAID, Immunopathol Lab, 7 Ctr Dr,Room 304,MSC 0760, Bethesda, MD 20892 USA. OI Morse, Herbert/0000-0002-9331-3705 NR 30 TC 5 Z9 5 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-2126 J9 LEUKEMIA RES JI Leuk. Res. PD AUG PY 2002 VL 26 IS 8 BP 739 EP 743 AR PII S0145-2126(01)00200-4 DI 10.1016/S0145-2126(01)00200-4 PG 5 WC Oncology; Hematology SC Oncology; Hematology GA 574RK UT WOS:000176903200007 PM 12191569 ER PT J AU Weisinger, HS Armitage, JA Jeffrey, BG Mitchell, DC Moriguchi, T Sinclair, AJ Weisinger, RS Salem, N AF Weisinger, HS Armitage, JA Jeffrey, BG Mitchell, DC Moriguchi, T Sinclair, AJ Weisinger, RS Salem, N TI Retinal sensitivity loss in third-generation n-3 PUFA-deficient rats SO LIPIDS LA English DT Article ID POLYUNSATURATED FATTY-ACIDS; DOCOSAHEXAENOIC ACID; ELECTRICAL RESPONSE; LIGHT DAMAGE; GUINEA-PIGS; A-WAVE; BRAIN; ELECTRORETINOGRAM; MEMBRANES; DIETS AB A previous study conducted in guinea pigs suggested that ingestion of diets high in EPA and DHA may result in suboptimal retinal function. The aim of the present study was to evaluate retinal function in pigmented (Long-Evans) rats, raised to a third generation on diets that were either deficient in n-3 PUFA or adequate (with the addition of DHA). Electroretinographic assessment employed full-field white flash stimulation. Photoreceptor responses were evaluated in terms of peak amplitudes and implicit times (a-wave, b-wave), intensity-response functions (Naka-Rushton), and the parameters of a model of transduction (P3). Retinal phospholipid FA composition was measured by capillary GLC. DHA levels were reduced by 55% in n-3-deficient animals compared with the n-3-adequate group, whereas the levels of docosapentaenoic acid n-6 were 44 times higher in n-3-deficient animals. The level of arachidonic acid was marginally higher (12.8%) in n-6-adequate animals. The n-3-deficient animals exhibited significantly reduced retinal sensitivity (a and S values were both affected by 0.29 log units) and increased b-wave implicit times compared with those fed the n-3-adequate diet. These data suggest that n-3 PUFA are required for development of retinal sensitivity, more so than other indices of retinal function assessed by current methods, such as maximal response amplitude. However, the benefit for retinal function of adding preformed DHA to diets already replete in n-3 PUFA remains unclear. C1 Univ Melbourne, Howard Florey Inst Expt Physiol & Med, Melbourne, Vic 3010, Australia. NIAAA, Lab Membrane Biochem & Biophys, NIH, Rockville, MD 20852 USA. Royal Melbourne Inst Technol, Dept Food Sci, Melbourne, Vic 3000, Australia. Flinders Univ S Australia, Flinders Med Ctr, Dept Paediat & Child Hlth, Adelaide, SA 5042, Australia. RP Weisinger, RS (reprint author), Univ Melbourne, Howard Florey Inst Expt Physiol & Med, Melbourne, Vic 3010, Australia. RI Armitage, James/A-8949-2012 NR 44 TC 33 Z9 33 U1 0 U2 1 PU AMER OIL CHEMISTS SOC A O C S PRESS PI CHAMPAIGN PA 1608 BROADMOOR DRIVE, CHAMPAIGN, IL 61821-0489 USA SN 0024-4201 J9 LIPIDS JI Lipids PD AUG PY 2002 VL 37 IS 8 BP 759 EP 765 DI 10.1007/s11745-002-0958-3 PG 7 WC Biochemistry & Molecular Biology; Nutrition & Dietetics SC Biochemistry & Molecular Biology; Nutrition & Dietetics GA 599ZL UT WOS:000178365400004 PM 12371746 ER PT J AU Pergolizzi, S Santacaterina, A De Renzis, C Settineri, N Gaeta, M Frosina, P Russi, EG Altavilla, G AF Pergolizzi, S Santacaterina, A De Renzis, C Settineri, N Gaeta, M Frosina, P Russi, EG Altavilla, G TI Older people with non small cell lung cancer in clinical stage IIIA and co-morbid conditions - Is curative irradiation feasible? Final results of a prospective study SO LUNG CANCER LA English DT Article DE older; lung; radiotherapy ID PATIENTS AGED 75; RADIATION-THERAPY; ELDERLY PATIENTS; RADIOTHERAPY; CARCINOMA; SURVIVAL; NSCLC; MANAGEMENT; TOXICITY; IMPACT AB Radiotherapy (RT) is widely used in the management of lung cancer but age-oriented randomized trials are lacking in older-unfit patients. We started a prospective study to evaluate the toxicity and efficacy of definitive RT in very old-unfit patients with non-small cell lung cancer (NSCLC) in clinical stage IIIA, according to AJCC 1988. Forty patients, age greater than or equal to 75 years, Karnofsky Performance status (KPS)greater than or equal to60, unfit to receive an aggressive combined treatment, were entered in the study. Each patient had one or more comorbidities, and the Charlson score was greater than two in 7/40. All patients were treated with radiation fields encompassing the primary tumor and grossly involved lymph nodes. A median radiation dose of 60 Gy/2 Gy day/5 days a week, was delivered. The 40 patients have been followed up, including those who died, for a potential median time of 4.6 years. As results, no treatment-related mortality, and clinically insignificant acute morbidity was recorded: in 28/40 cases a mild esophagitis occurred. Two patients showed a clinical radiation pneumonitis (RP). Late normal tissue damage was represented by lung fibrosis (40/40 patients). The treatment was efficacy since each patient obtained some clinical benefit from it. Median survival (MS) was 19 months (range 5-68); the 3 and 5-year actuarial survival was 18 and 12%, respectively. In conclusion, we think that older patients with concomitant illness can be submitted to curative 'involved field' irradiation and the results observed in this trial encourage to use curative RT in older subjects with local-regionally advanced NSCLC and co-morbid condition. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Messina, Div Radiat Therapy, Dept Radiol Sci, Messina, Italy. Natl Canc Inst, Genoa, Italy. Univ Messina, Div Radiol, Dept Radiol Sci, Messina, Italy. Hosp S Croce & Carle, Dept Radiat Therapy, Cuneo, Italy. Univ Messina, Div Med Oncol, Dept Human Pathol, Messina, Italy. RP Pergolizzi, S (reprint author), Via Salita Ogliastri 16, I-98100 Messina, Italy. NR 30 TC 22 Z9 22 U1 0 U2 6 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0169-5002 J9 LUNG CANCER-J IASLC JI Lung Cancer PD AUG PY 2002 VL 37 IS 2 BP 201 EP 206 AR PII S0169-5002(02)00038-7 DI 10.1016/S0169-5002(02)00038-7 PG 6 WC Oncology; Respiratory System SC Oncology; Respiratory System GA 586XD UT WOS:000177610700012 PM 12140143 ER PT J AU Subramanian, S Devasahayam, N Murugesan, R Yamada, K Cook, J Taube, A Mitchell, JB Lohman, JAB Krishna, MC AF Subramanian, S Devasahayam, N Murugesan, R Yamada, K Cook, J Taube, A Mitchell, JB Lohman, JAB Krishna, MC TI Single-point (constant-time) imaging in radiofrequency Fourier transform electron paramagnetic resonance SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE EPR imaging; Fourier transform; single point; constant time; functional imaging ID T-1 ENHANCEMENT SPRITE; HIGH-RESOLUTION; ESR SPECTROMETER; SPECTROSCOPY; EPR; SENSITIVITY; MICROSCOPY; RADIATION; SOLIDS; MICE AB This study describes the use of the single-point imaging (SPI) modality, also known as constant-time imaging (CTI), in radiofrequency (RF) Fourier transform (FT) electron paramagnetic resonance (EPR). The SPI technique, commonly used for high-resolution solid-state nuclear magnetic resonance (NMR) imaging, has been successfully applied to 2D and 3D RF-FT-EPR imaging of phantoms containing narrow-line EPR spin probes. The SPI scheme is essentially a phase-encoding technique that operates by acquiring a single data point in the free induction decay (FID) after a fixed delay (phase-encoding time), following the pulsed RF excitation, in the presence of static magnetic field gradients. Since the phase-encoding time remains constant for a given image data set, the spectral information is automatically deconvolved, providing well-resolved pure spatial images. Therefore, images obtained using SPI are artifact-free and the resolution is not significantly limited by the line width, compared to the images obtained using the conventional filtered back-projection (FBP) scheme, suggesting that the SPI modality may have advantages for EPR imaging of large objects. In this work the advantages and limitations of SPI as compared to FBP are investigated by imaging suitable phantom objects. Although SPI takes longer to perform than the FBP method, optimization of the data collection scheme may increase the temporal resolution, rendering this technique suitable for in vivo studies. Spectral information can also be extracted from a series of SPI images that are generated as a function of the delay from the excitation pulse. C1 NCI, NIH, Radiat Biol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. Bruker Med Ltd, Coventry, W Midlands, England. RP Krishna, MC (reprint author), NCI, NIH, Radiat Biol Branch, Ctr Canc Res, Bldg 10,Room B3 B69, Bethesda, MD 20892 USA. RI Yamada, Ken-ichi/E-6318-2012 NR 40 TC 53 Z9 53 U1 1 U2 8 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD AUG PY 2002 VL 48 IS 2 BP 370 EP 379 DI 10.1002/mrm.10199 PG 10 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 577UK UT WOS:000177080300018 PM 12210946 ER PT J AU Baxter, LL Pavan, WJ AF Baxter, LL Pavan, WJ TI The oculocutaneous albinism type IV gene Matp is a new marker of pigment cell precursors during mouse embryonic development SO MECHANISMS OF DEVELOPMENT LA English DT Article DE AIM-1; Matp; melanocyte; neural crest; melanoblast; melanoma; Mitf; oculocutaneous albinism; retinal pigmented epithelium; underwhite; pigment cell; tyrosinase; dopachrome tautomerase; melanogenic enzyme; hypopigmentation ID CREST-DERIVED MELANOCYTES; TRANSCRIPTION FACTOR; MICROPHTHALMIA; MUTATIONS; MITF; MELANOMA; PROMOTER; PROTEIN; GROWTH; LOCUS AB Expression profile analysis demonstrated that the expression of membrane-associated transporter protein (MATP) varied similarly to the melanogenic enzymes dopachrome tautomerase (DCT) and tyrosinase related protein 1 (TYRP1) (Proc. Nail Acad. Sci. USA (2002) in press). Mutations in MATP result in pigmentation alterations in mice (underwhite, uw), in medaka (b-locus), and in man (Oculocutaneous Albinism Type 4, OCA4) (Nat. Genet. 28 (2001) 381; Am. J. Hum. Genet. 69 (2001) 98 1). Consistent with MATP acting in a pigment cell autonomous manner, in situ hybridization analysis demonstrated expression of murine Matp in the presumptive retinal pigmented epithelium starting at E9.5, and in neural crest-derived melanoblasts starting at E10.5. Matp expression is reduced in embryos mutated for microphthalmia-associated transcription factor (Mitf) (Cell 74 (1993) 395; J. Biol. Chem. 268 (1993) 20687), suggesting Mitf regulates Matp expression. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 NHGRI, Mouse Embryol Sect, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. RP Pavan, WJ (reprint author), NHGRI, Mouse Embryol Sect, Genet Dis Res Branch, NIH, 49,Convent Dr,Bldg 49,Room 4A82, Bethesda, MD 20892 USA. NR 17 TC 23 Z9 25 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0925-4773 J9 MECH DEVELOP JI Mech. Dev. PD AUG PY 2002 VL 116 IS 1-2 BP 209 EP 212 AR PII S0925-4773(02)00130-2 DI 10.1016/S0925-4773(02)00130-2 PG 4 WC Developmental Biology SC Developmental Biology GA 582CW UT WOS:000177333000026 PM 12128226 ER PT J AU Warren, JL AF Warren, JL TI Use of the SEER-Medicare Data for Cancer-Related Health Services Research - Data sources, analytic issues, and methods - Preface SO MEDICAL CARE LA English DT Editorial Material C1 NCI, Appl Res Program, DCCPS, Bethesda, MD 20892 USA. RP Warren, JL (reprint author), NCI, Appl Res Program, DCCPS, Execut Plaze N,Room 4005,6130 Execut Blvd,MSC 734, Bethesda, MD 20892 USA. NR 0 TC 18 Z9 18 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 1 EP 2 PG 2 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200001 ER PT J AU Warren, JL Klabunde, CN Schrag, D Bach, PB Riley, GF AF Warren, JL Klabunde, CN Schrag, D Bach, PB Riley, GF TI Overview of the SEER-Medicare data - Content, research applications, and generalizability to the United States elderly population SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE SEER Program; Medicare; cancer; health services research; epidemiology ID HEALTH MAINTENANCE ORGANIZATIONS; BREAST-CANCER; CLAIMS DATA; COLORECTAL-CANCER; LUNG-CANCER; OLDER WOMEN; PROSTATE-CANCER; COLON-CANCER; ADJUVANT CHEMOTHERAPY; HOSPITAL VOLUME AB BACKGROUND. The Surveillance, Epidemiology and End,Results (SEER)-Medicare-linked database combines clinical information from population-based cancer, registries with claims information from the Medicare program. The use of this database to study cancer screening, treatment, outcomes, and costs has grown in recent years. RESEARCH DESIGN. This paper provides an overview of the SEER-Medicare files for investigators interested in using these data for epidemiologic and health services research. The overview includes a description of the linkage of SEER and Medicare data and the files included as part of SEER-Medicare. The paper also describes the types of research projects that have been undertaken using the SEER-Medicare data. The overview concludes with a comparison of selected characteristics of elderly persons residing in the SEER areas to the US total aged. RESULTS. The paper identifies a number of potential uses of. the SEER-Medicare data. The comparison of the elderly population in SEER to the US total shows that in the SEER areas areas there are a lower percentage of white and individuals living in poverty, and persons and, a higher percentage of urban-dwellers than the US total. Elderly persons in the SEER regions also have higher rates of HMO enrollment and lower rates of cancer mortality. CONCLUSIONS. The SEER-Medicare data are a unique resource that can be used for a variety of health services research projects. Although there are some differences between the elderly residing in the SEER areas and the US total, the SEER-Medicare data offer a large population-based cohort that can be used to longitudinally track care for persons over the course of cancer p, diagnosis, treatment, and follow-up. C1 NCI, Appl Res Program, DCCPS, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, Hlth Outcomes Res Grp, New York, NY 10021 USA. Ctr Medicare Serv, Baltimore, MD USA. Ctr Medicaid Serv, Baltimore, MD USA. RP Warren, JL (reprint author), NCI, Appl Res Program, DCCPS, Execut Plaza N,Room 4005,6130 Execut Blvd,MSC 734, Bethesda, MD 20892 USA. NR 67 TC 751 Z9 753 U1 11 U2 30 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 3 EP 18 PG 16 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200002 ER PT J AU Bach, PB Guadagnoli, E Schrag, D Schussler, N Warren, JL AF Bach, PB Guadagnoli, E Schrag, D Schussler, N Warren, JL TI Patient demographic and socioeconomic characteristics in the SEER-Medicare database - Applications and limitations SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE SEER program; Medicare; demography; social class; racial stocks ID CENSUS-DATA; CANCER; MORTALITY; SURVIVAL; TRANSPLANTATION; ACCURACY; DIALYSIS; OUTCOMES; TRENDS AB Users of the linked SEER-Medicare database commonly perform analyses that focus on the complex interactions among patient characteristics, cancer treatments, and outcomes. The authors review the source. and scope of the patient-specific data elements, with a focus on three domains-demographic characteristics, socioeconomic characteristics, and survival status. They offer some concrete recommendations regarding the use of these data elements. In particular, they describe analyses that provide an estimate of the accuracy of the sex and age variables, and raise some cautionary notes about race and ethnicity variables. The authors describe the available measures of socioeconomic status, and recommend, with some caveats, the use of median income measures as a proxy for socioeconomic status. Finally, they describe the available data on date of death, and explain why confidence in these measures is justified. C1 Mem Sloan Kettering Canc Ctr, Hlth Outcomes Res Grp, Dept Epidemiol & Biostat, New York, NY 10021 USA. Harvard Univ, Sch Med, Dept Hlth Care Policy, Boston, MA 02115 USA. Informat Management Serv Inc, Silver Spring, MD USA. NCI, Appl Res Branch, Bethesda, MD USA. RP Bach, PB (reprint author), Mem Sloan Kettering Canc Ctr, Hlth Outcomes Res Grp, Dept Epidemiol & Biostat, 1275 York Ave,Box 221, New York, NY 10021 USA. NR 16 TC 164 Z9 165 U1 2 U2 8 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 19 EP 25 PG 7 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200003 ER PT J AU Klabunde, CN Warren, JL Legler, JM AF Klabunde, CN Warren, JL Legler, JM TI Assessing comorbidity using claims data - An overview SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE comorbidity; claims data; administrative data; SEER; Medicare ID ICD-9-CM ADMINISTRATIVE DATA; BREAST-CANCER SURVIVAL; PROSTATE-CANCER; CO-MORBIDITY; HOSPITAL MORTALITY; MEDICARE PATIENTS; RISK ADJUSTMENT; CHRONIC DISEASE; ELDERLY WOMEN; PRIMARY-CARE AB Comorbidity, additional disease beyond the condition under study that increases a patient's total burden of illness, is one dimension of health status. For investigators working with observational data obtained from administrative databases, comorbidity assessment may be a useful and important means of accounting for differences in patients' underlying health status. There are multiple ways of measuring comorbidity. This paper provides an overview of current approaches to and issues in assessing comorbidity using claims data, with a particular focus on established indices and the SEER-Medicare database. in addition, efforts to improve measurement of comorbidity using claims data are described, including augmentation of claims data with medical record, patient self-report, or health services utilization data; incorporation of claims data from sources other than inpatient claims; and exploration of alternative conditions, indices, or ways of grouping conditions. Finally, caveats about claims data and areas for future research in claims-based comorbidity assessment are discussed. Although the use of claims databases such as SEER-Medicare for health services and outcomes research has become increasingly common, investigators must be cognizant of the limitations of comorbidity measures derived from these data sources in capturing and controlling for differences in patient health status. The assessment of comorbidity using claims data is a complex and evolving area of investigation. C1 NCI, Appl Res Program, Hlth Serv & Econ Branch, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. NCI, Stat Res & Applicat Branch, Surveillance Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. RP Klabunde, CN (reprint author), NCI, Appl Res Program, Hlth Serv & Econ Branch, Div Canc Control & Populat Sci, Execut Plaza N,Room 4005,6130 Execut Blvd, Bethesda, MD 20892 USA. NR 70 TC 213 Z9 215 U1 6 U2 13 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 26 EP 35 PG 10 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200004 ER PT J AU Freeman, JL Klabunde, CN Schussler, N Warren, JL Virnig, BA Cooper, GS AF Freeman, JL Klabunde, CN Schussler, N Warren, JL Virnig, BA Cooper, GS TI Measuring breast, colorectal, and prostate cancer screening with Medicare claims data SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE health services utilization; cancer screening; Medicare; SEER ID OLDER WOMEN; MAMMOGRAPHY USE; CALIFORNIA MEDICARE; ELDERLY WOMEN; SURVEILLANCE; POPULATION; DIAGNOSIS; STAGE; CARE AB BACKGROUND. Evaluating the use and effectiveness of cancer screening is an important component of cancer control programs. Medicare claims may be a useful source of data when screening older populations, but they are limited in terms of completeness and the ability to distinguish screening tests from those provided for diagnosis or surveillance. RESEARCH DESIGN. A review of the major screening modalities for breast, colorectal, and prostate cancer, Medicare's policies for covering these tests, and the procedure codes used to identify them in Medicare claims. RESULTS. Although Medicare's coverage has been extended to include screening mammography, colonoscopy, sigmoidoscopy, fecal occult blood tests, double-contrast barium enema, and prostate-specific antigen tests, providers have been slow to adopt the corresponding screening codes. CONCLUSION. Challenges persist in measuring screening use, and innovative approaches are required to distinguish screening tests from diagnostic and follow-up evaluations. C1 Univ Texas, Med Branch, Sealy Ctr Aging, Galveston, TX 77555 USA. NCI, Appl Res Program, Bethesda, MD USA. Informat Management Serv Inc, Silver Spring, MD USA. Univ Minnesota, Sch Publ Hlth, Div Hlth Serv Res & Policy, Minneapolis, MN 55455 USA. Case Western Reserve Univ, Dept Med, Cleveland, OH 44106 USA. Case Western Reserve Univ, Canc Res Ctr, Cleveland, OH 44106 USA. RP Freeman, JL (reprint author), Univ Texas, Med Branch, Sealy Ctr Aging, 301 Univ Blvd, Galveston, TX 77555 USA. NR 23 TC 45 Z9 45 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 36 EP 42 PG 7 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200005 ER PT J AU Cooper, GS Virnig, B Klabunde, CN Schlussler, N Freeman, J Warren, JL AF Cooper, GS Virnig, B Klabunde, CN Schlussler, N Freeman, J Warren, JL TI Use of SEER-Medicare data for measuring cancer surgery SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE breast neoplasms; colorectal neoplasms; endometrial neoplasms; lung neoplasms; pancreatic neoplasms; prostatic neoplasms; Medicare ID CLAIMS DATA; ACCURACY; VOLUME AB BACKGROUND. The accuracy and completeness of the SEER-Medicare data for measuring cancer-related therapy have not been extensively evaluated. OBJECTIVES. To investigate the best method for measuring cancer-related surgery among patients in the SEER-Medicare database. SUBJECTS. A total of 149,970 incident cases of breast, colorectal, endometrial, lung, pancreatic, and prostate cancer diagnosed between 1991 and 1993. MEASURES. The most invasive surgical procedure identified through Medicare's inpatient, physician, and hospital outpatient claims was compared with corresponding data from the SEER files. RESULTS. Agreement between the SEER and Medicare files was generally highest for resection and radical surgery (eg, kappa 0.70-0.90). While there was less agreement regarding no surgical therapy and biopsy individually, the concordance of the two sources in excluding cancer-directed surgery was high. Compared with inpatient data alone, using the combined inpatient, physician, and outpatient data increased concordance between SEER and Medicare for less invasive procedures. CONCLUSIONS. The agreement of SEER and Medicare data appears to be good for major surgical procedures and for excluding persons who did not undergo cancer-directed surgery. Both the SEER and the Medicare data captured a small number of surgeries not reported in the other file. Therefore, where possible, using both data sources will enhance identification of surgeries. Because the analysis was performed with linked data, the accuracy of surgical claims in Medicare data alone cannot be assessed. C1 Univ Minnesota, Sch Publ Hlth, Div Hlth Serv Res & Policy, Minneapolis, MN 55455 USA. NCI, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Informat Management Serv Inc, Silver Spring, MD USA. Univ Texas, Med Ctr, Div Geriatr Med, Galveston, TX 77555 USA. RP Cooper, GS (reprint author), Univ Hosp Cleveland, Div Gastroenterol, 11100 Euclid Ave, Cleveland, OH 44106 USA. NR 10 TC 163 Z9 164 U1 1 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 43 EP 48 PG 6 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200006 ER PT J AU Virnig, BA Warren, JL Cooper, GS Klabunde, CN Schussler, N Freeman, J AF Virnig, BA Warren, JL Cooper, GS Klabunde, CN Schussler, N Freeman, J TI Studying radiation therapy using SEER-Medicare-linked data SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE radiation therapy; cancer care; Medicare; elderly; brachytherapy ID BREAST-CANCER; OLDER WOMEN; STAGE AB BACKGROUND. Numerous studies have used the SEER or Medicare data to assess the use of radiation therapy. However, the completeness of these data has not been evaluated. METHODS. Using linked SEER-Medicare data, a cohort was created that included incident cases of breast, endometrial, lung, prostate, or rectal cancer from 1991 to 1996 in persons 65 years and older. For each of these cases, use of radiation therapy as an initial course of treatment reported from the SEER data was compared with use of radiation therapy in the months following diagnosis, as determined from the Medicare claims. RESULTS. Overall, there was a high agreement between SEER and Medicare sources for use of radiation therapy. Agreement was 88% for lung cancer, 93% for prostate cancer, 94% for rectal and breast cancers, and 95% for endometrial cancer. Within the Medicare files, radiation claims were found in MedPAR Physician Supplier Part B and Outpatient files. CONCLUSIONS. There is a high level of agreement between SEER and Medicare reporting of radiation treatments after a cancer diagnosis, suggesting that either source can be used to assess radiation-related treatment patterns. However, for the most complete ascertainment of radiation use, both SEER and Medicare sources should be used. C1 Univ Minnesota, Sch Publ Hlth, Div Hlth Serv Res & Policy, Minneapolis, MN 55455 USA. NCI, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Case Western Reserve Univ, Dept Med, Cleveland, OH 44106 USA. Informat Management Serv Inc, Silver Spring, MD USA. Univ Texas, Med Ctr, Div Geriatr Med, Galveston, TX 77555 USA. RP Virnig, BA (reprint author), Univ Minnesota, Sch Publ Hlth, Div Hlth Serv Res & Policy, 420 Delaware St SE,MMC 97, Minneapolis, MN 55455 USA. NR 9 TC 176 Z9 177 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 49 EP 54 PG 6 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200007 ER PT J AU Warren, JL Harlan, LC Fahey, A Virnig, BA Freeman, JL Klabunde, CN Cooper, GS Knopf, KB AF Warren, JL Harlan, LC Fahey, A Virnig, BA Freeman, JL Klabunde, CN Cooper, GS Knopf, KB TI Utility of the SEER-Medicare data to identify chemotherapy use SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE SEER Program; Medicare; cancer; chemotherapy; claims analysis ID NEGATIVE BREAST-CANCER; PRACTICE GUIDELINES AB BACKGROUND. Medicare claims include codes for chemotherapy administration and specific drugs given, and researchers are increasingly using these data to measure the use of chemotherapy. However, the validity and completeness of these data as a source of information has not been established. OBJECTIVES. This analysis is intended to assess the utility of the Medicare claims to capture chemotherapy use. METHODS. Persons with breast, colorectal, and ovarian cancer were identified from the linked SEER-Medicare data. Their Medicare claims were reviewed to determine if there were any bills for chemotherapy, and if so, if there were claims for specific agents. This information was compared with data on the first course of treatment obtained from hospitals and treating physicians by the SEER registries through an NCI-supported Patterns of Care Studies (POC). Agreement was measured using kappa statistics. The sensitivity of the Medicare claims to capture chemotherapy, as reported from the POC data, was also measured. An additional comparison assessed the agreement between the two data sources concerning which specific drugs had been given. RESULTS. For all of the cancers, there was a high level of agreement between the Medicare claims and the POC data regarding whether or not the patient had received chemotherapy (kappa greater than or equal to0.73). The sensitivity of the Medicare data to determine if a person had received chemotherapy was high (greater than or equal to88%). In cases where the Medicare claim included a code for a specific drug, there high agreement between Medicare and POC about the specific drug given in breast and colorectal cancers, although the agreement was lower for ovarian cancers. The sensitivity of the Medicare claims to identify specific agents varies by cancer type. CONCLUSIONS. The Medicare claims can be used to identify which persons are receiving chemotherapy. The utility of Medicare data to measure treatment with specific agents varies by cancer type and specific agent. For some cancers, it is possible to use these claims to assess use of specific drugs, while for other drugs the data are limited. C1 NCI, Appl Res Program, DCCPS, Bethesda, MD 20892 USA. Informat Management Serv Inc, Silver Spring, MD USA. Univ Minnesota, Sch Publ Hlth, Div Hlth Serv Res & Policy, Minneapolis, MN 55455 USA. Univ Texas, Med Branch, Sealy Ctr Aging, Galveston, TX 77550 USA. Case Western Reserve Univ, Dept Med, Cleveland, OH 44106 USA. Case Western Reserve Univ, Canc Res Ctr, Cleveland, OH 44106 USA. RP Warren, JL (reprint author), NCI, Appl Res Program, DCCPS, Execut Plaza N,Room 4005,6130 Execut Blvd,MSC 734, Bethesda, MD 20892 USA. NR 13 TC 224 Z9 225 U1 0 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 55 EP 61 PG 7 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200008 ER PT J AU Potosky, AL Warren, JL Riedel, ER Klabunde, CN Earle, CC Begg, CB AF Potosky, AL Warren, JL Riedel, ER Klabunde, CN Earle, CC Begg, CB TI Measuring complications of cancer treatment using the SEER-Medicare data SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE neoplasms; outcome assessment; adverse effects; acute complications; chronic complications; measurement; SEER; Medicare ID ADMINISTRATIVE DATA; PROSTATE-CANCER; CLAIMS DATA; OUTCOMES; MORTALITY; ACCURACY; SURGERY; VOLUME; CARE AB BACKGROUND. The linkage of SEER registry data with Medicare claims allows the longitudinal tracking of health care and outcomes for patients after a cancer diagnosis. One category of outcomes amenable to research using Medicare claims is complications of cancer treatments: the unintentional, adverse side effects or sequelae of interventions used to treat or palliate cancer patients. RESEARCH DESIGN. The authors review some of the methods and limitations of using Medicare claims to identify both acute and chronic complications of cancer treatments, and present an original analysis comparing survey based and claims-based complications following radical prostatectomy for prostate cancer to illustrate some of the potential limitations inherent in using claims for this purpose. RESULTS. Utility of the Medicare claims for identifying postdischarge complications varies by the patient type, the initial treatment used, and any subsequent treatment of complications. For patients undergoing surgical interventions, Medicare claims can be used to identify most acute inpatient complications. However, claims data cannot be used as effectively in the long-term to capture chronic complications, particularly when the complication does not consistently prompt an intervention. CONCLUSION. Researchers who use the SEER-Medicare-linked database to assess long-term complications of cancer treatments should exercise caution when designing and interpreting studies. Ideally, for studies of most chronic complications of cancer care, validation studies similar to the one performed here would provide valuable additional evidence to assess the credibility of conclusions based on claims data. C1 NCI, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. Harvard Univ, Dana Farber Canc Inst, Boston, MA 02115 USA. RP Potosky, AL (reprint author), NCI, Appl Res Program, Div Canc Control & Populat Sci, EPN Room 4005,6130 Execut Blvd,MSC 7344, Bethesda, MD 20892 USA. NR 17 TC 71 Z9 71 U1 3 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 62 EP 68 PG 7 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200009 ER PT J AU Nattinger, AB Schapira, MM Warren, JL Earle, CC AF Nattinger, AB Schapira, MM Warren, JL Earle, CC TI Methodological issues in the use of administrative claims data to study surveillance after cancer treatment SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE cancer surveillance; Medicare claims data; Tumor Registry data ID BREAST-CANCER; MEDICARE BENEFICIARIES; COLORECTAL-CANCER; FOLLOW-UP; MAMMOGRAPHY; CARE AB BACKGROUND. Administrative claims databases linked to tumor registry data provide an excellent opportunity for studying the follow-up care of cancer patients. RESEARCH DESIGN. Methodological principles of study design are reviewed, using examples drawn from the recently published literature. RESULTS. Most follow-up care is outpatient-based. Therefore, in studies using Medicare claims, the patient should be eligible,for Medicare Part B and should not be enrolled in a Medicare HMO. In studies of surveillance testing, it may be appropriate to exclude subjects who are near death, in a hospice, or in whom a new or recurrent cancer develops. The definition of the period for measurements of surveillance testing requires consideration, as does periodicity in patterns of testing. Several analytic methods can be employed, from proportions undergoing testing to survival analysis methods. Measurement of 'surveillance tests' among control subjects (those without cancer) may be useful for comparison with cancer patients. To date, administrative claims data have been most useful in studies investigating the quality of, and disparities in, cancer care. CONCLUSION. With appropriate attention to methodological issues, linked tumor registry and administrative databases can provide important insights into the quality of survivorship care for cancer. C1 Med Coll Wisconsin, Dept Med, Milwaukee, WI 53226 USA. NCI, Appl Res Program, Bethesda, MD 20892 USA. Dana Farber Canc Inst, Dept Adult Oncol, Boston, MA 02115 USA. RP Nattinger, AB (reprint author), Div Gen Internal Med, FEC Med Off Bldg,Suite 4200,9200 W Wisconsin Ave, Milwaukee, WI 53226 USA. NR 13 TC 21 Z9 21 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 69 EP 74 PG 6 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200010 ER PT J AU Earle, CC Nattinger, AB Potosky, AL Lang, K Mallick, R Berger, M Warren, JL AF Earle, CC Nattinger, AB Potosky, AL Lang, K Mallick, R Berger, M Warren, JL TI Identifying cancer relapse using SEER-Medicare data SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE cancer; health services research; administrative data ID CHEMOTHERAPY AB INTRODUCTION. Tumor registries capture valid information at the time of cancer diagnosis, but often do not conduct longitudinal follow-up evaluations. However, investigators may be interested in questions relating to subsequent relapsed disease. Linking administrative data to registry data, as in the creation of the SEER (Surveillance, Epidemiology, and End Results) and Medicare data set, can provide the ability to infer the occurrence of relapse in selected situations. METHODS. The authors created different algorithms to detect relapse of acute myelogenous leukemia (AML). A retrospective cohort of patients with AML was identified, and both their billing data and medical records were obtained. The algorithms were then applied to the billing data, the results were compared with medical record review. RESULTS. Eighty-nine patients were identified, of whom 22 were treated for relapsed AML. The sensitivity of the best algorithm for detecting relapse was 86%, and the specificity 99%, with a positive predictive value of 95% and a negative predictive value of 96%. CONCLUSIONS. Identification of relapse from SEER-Medicare data using clinical algorithms is feasible for cancers where a majority of patients receive treatment for relapse, without a "watch and wait" strategy, and where that treatment is with a modality that can be detected in billing data (ie, intravenous chemotherapy, radiation, surgery, or all three). Optimal analytic situations are ones in which the investigator is mostly interested in positive predictive value, less interested in sensitivity, and wants to evaluate outcomes among those patients who receive treatment for their relapsed disease. However, the accuracy of such an approach for cancers other than AML has not yet been established. C1 Dana Farber Canc Inst, Ctr Outcomes & Policy Res, Dept Adult Oncol, Boston, MA 02115 USA. Med Coll Wisconsin, Madison, WI USA. NCI, Appl Res Program, Bethesda, MD USA. Boston Hlth Econ, Boston, MA USA. Wyeth Ayerst Res, Madison, NJ USA. RP Earle, CC (reprint author), Dana Farber Canc Inst, Ctr Outcomes & Policy Res, Dept Adult Oncol, 44 Binney St,454-STE 21-24, Boston, MA 02115 USA. NR 9 TC 59 Z9 60 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 75 EP 81 PG 7 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200011 ER PT J AU Baldwin, LM Adamache, W Klabunde, CN Kenward, K Dahlman, C Warren, JL AF Baldwin, LM Adamache, W Klabunde, CN Kenward, K Dahlman, C Warren, JL TI Linking physician characteristics and Medicare claims data - Issues in data availability, quality, and measurement SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE physician characteristics; Medicare; claims data ID BREAST-CANCER-TREATMENT; PRIMARY-CARE; ELDERLY PATIENTS; HOSPITAL VOLUME; PREVENTIVE CARE; SURGEON VOLUME; JUDGMENTS; OUTCOMES; WOMEN AB BACKGROUND. Increasingly, investigators are using administrative databases to answer research questions requiring physician characteristics information. This article provides a roadmap for investigators who use Medicare data to answer such questions, focusing on use of the Surveillance, Epidemiology, and End Results (SEER)-Medicare files. METHODS. Three data sources that can be linked to identify physician characteristics-Medicare claims, the Unique Physician Identification Number (UPIN) Registry, and the American Medical Association (AMA) Masterfile-were examined for data availability, linkage rates, and quality. These databases also were used to explore measurement issues regarding physician specialty and practice volume. RESULTS. Over 98 percent of UPINs identified from the Medicare claims could be linked with both the AMA Masterfile and the UPIN Registry. The AMA Masterfile is the best source of sociodemographic and medical training information; the Medicare claims are the best source of practice ZIP code; and the UPIN Registry is the best source of practice organization data. The operationalization of variables such as physician specialty and practice volume is dependent on the specific research question that is being addressed. CONCLUSION. Administrative databases, such as SEER-Medicare data linked to AMA Masterfile or UPIN Registry data, are an important resource for investigators interested in assessing the relationship between physicians' personal and practice characteristics and the content or outcomes of clinical care. C1 Univ Washington, Dept Family Med, Seattle, WA 98105 USA. Hlth Econ Res Inc, Waltham, MA USA. NCI, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Amer Med Assoc, Div Survey & Data Resources, Chicago, IL 60610 USA. CHD Res Associates, Columbia, MD USA. RP Baldwin, LM (reprint author), Univ Washington, Dept Family Med, Suite 308,4225 Roosevelt Way, Seattle, WA 98105 USA. NR 19 TC 76 Z9 76 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 82 EP 95 PG 14 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200012 ER PT J AU Schrag, D Bach, PB Dahlman, C Warren, JL AF Schrag, D Bach, PB Dahlman, C Warren, JL TI Identifying and measuring hospital characteristics using the SEER-Medicare data and other claims-based sources SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE hospital characteristics; SEER-Medicare; administrative data; research methods ID QUALITY; CARE; VOLUME; CANCER; MORTALITY; SURGERY AB The context in which health care is delivered may have an impact on the outcomes and processes of care. Researchers analyzing claims data that reflect care rendered at many institutions may wish to identify hospital characteristics such as bed size, teaching status, degree of specialization, and case volume to evaluate their influence on outcomes. The authors describe the resources available from administrative data for characterizing hospitals. The article focuses on analyses using the SEER-Medicare-linked data, but has direct relevance for investigators working with other sources of claims data. First, the authors focus on assessment of hospital case volume, which is the principal hospital attribute that can be obtained directly from the SEER-Medicare data files. Second, they assess the information regarding hospital characteristics that can be gleaned from the American Hospital Association (AHA) survey and the annual cost reports submitted to the Center for Medicare and Medicaid Services (CMS) by all certified hospitals. From a comparison of the responses of 4,434 hospitals to 1994 AHA and CMS surveys, the authors conclude that the two data sources contain very similar information, but that for most investigators, use of data from CMS cost reports will be preferable to use of AHA data because of higher survey-completion rates and public availability. C1 Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, Hlth Outcomes Res Grp, New York, NY 10021 USA. NCI, Appl Res Branch, Bethesda, MD 20892 USA. RP Schrag, D (reprint author), Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, Hlth Outcomes Res Grp, Box 221,1275 York Ave, New York, NY 10021 USA. NR 10 TC 34 Z9 34 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 96 EP 103 PG 8 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200013 ER PT J AU Brown, ML Riley, GF Schussler, N Etzioni, R AF Brown, ML Riley, GF Schussler, N Etzioni, R TI Estimating health care costs related to cancer treatment from SEER-Medicare data SO MEDICAL CARE LA English DT Article; Proceedings Paper CT SEER-Medicare Workshop CY NOV, 2000 CL BETHESDA, MARYLAND DE SEER; Medicare; cancer; cost; expenditures ID COLORECTAL-CANCER; MASTECTOMY; SERVICES; DISEASE; IMPACT AB BACKGROUND. Cancer-specific medical care costs are used by health service researchers, medical decision analysts, and health care policymakers. The SEER-Medicare database is a unique data resource that makes it possible to derive incidence- and prevalence-based estimates of cancer-related medical care costs by site and stage of disease, by treatment approach, and for age and gender strata for individuals older than 65 years. OBJECTIVES. This paper describes the cost-related data available in the SEER-Medicare database, and discusses techniques and methods that have been used to derive various cost estimates from these data. The limitations of SEER-Medicare data as a source of cost estimates are also discussed., RESULTS. Examples of cost estimates for colorectal and breast cancer derived from SEER-Medicare are presented, including estimates of incidence-based cost (average cost per patient) by the initial, terminal, and continuing care phases of cancer treatment. Estimates of cancer-related treatment costs, costs by type of treatment, and long-term costs are presented, as are prevalence-based costs (aggregate Medicare and national expenditures) by cancer type. C1 NCI, Hlth Serv & Econ Branch, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Ctr Medicare & Medicaid Serv, Baltimore, MD USA. IMS Inc, Silver Spring, MD USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. RP Brown, ML (reprint author), NCI, Hlth Serv & Econ Branch, Appl Res Program, Div Canc Control & Populat Sci, EPN-4005,6130 Execut Blvd, Bethesda, MD 20892 USA. NR 23 TC 212 Z9 214 U1 1 U2 8 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD AUG PY 2002 VL 40 IS 8 SU S BP 104 EP 117 PG 14 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 581NY UT WOS:000177300200014 ER PT J AU Stabler, SP Steegborn, C Wahl, MC Oliveriusova, J Kraus, JP Allen, RH Wagner, C Mudd, SH AF Stabler, SP Steegborn, C Wahl, MC Oliveriusova, J Kraus, JP Allen, RH Wagner, C Mudd, SH TI Elevated plasma total homocysteine in severe methionine adenosyltransferase I/III deficiency SO METABOLISM-CLINICAL AND EXPERIMENTAL LA English DT Article ID CYSTATHIONINE BETA-SYNTHASE; S-ADENOSYLMETHIONINE; FOLATE-DEFICIENCY; ISOLATED HYPERMETHIONINEMIA; CEREBROSPINAL-FLUID; INBORN-ERRORS; COBALAMIN; SERUM; RAT; METABOLISM AB Abnormal elevation of plasma methionine may result from several different genetic abnormalities, including deficiency of cystathionine beta-synthase (CBS) or of the isoenzymes of methionine adenosyltransferase (MAT) I and III expressed solely in nonfetal liver (MAT I/III deficiency). Classically, these conditions have been distinguished most readily by the presence or absence, respectively, of elevated plasma free homocystine, detected by amino acid chromatography in the former condition, but absent in the latter. During the present work, we have assayed methionine, S-adenosylmethionine, S-adenosylhomocysteine, total homocysteine (tHcy), cystathionine, N-methylglycine (sarcosine), and total cysteine (tCys) in groups of both MAT I/III- and CBS-deficient patients to provide more evidence as to their metabolite patterns. Unexpectedly, we found that MAT I/III-deficient patients with the most markedly elevated levels of plasma methionine also had elevations of plasma tHcy and often mildly elevated plasma cystathionine. Evidence is presented that methionine does not inhibit cystathionine beta-synthase, but does inhibit cystathionine gamma-lyase. Mechanisms that may possibly underlie the elevations of plasma tHcy and cystathionine are discussed. The combination of elevated methionine plus elevated tHcy may lead to the mistaken conclusion that an MAT I/III-deficient patient is instead CBS-deficient. Less than optimal management is then a real possibility. Measurements of plasma cystathionine, S-adenosylmethionine, and sarcosine should permit ready distinction between the 2 conditions in question, as well as be useful in several other situations involving abnormalities of methionine and/or homocysteine derivatives. Copyright 2002, Elsevier Science (USA). All rights reserved. C1 Univ Colorado, Hlth Sci Ctr, Dept Med, Denver, CO 80262 USA. Univ Colorado, Hlth Sci Ctr, Dept Pediat, Denver, CO 80262 USA. Univ Colorado, Hlth Sci Ctr, Dept Cellular & Struct Biol, Denver, CO 80262 USA. Max Planck Inst Biochem, Planegg Martinsried, Germany. Vanderbilt Univ, Dept Biochem, Nashville, TN 37232 USA. Vanderbilt Univ, Clin Nutr Res Unit, Nashville, TN 37232 USA. Dept Vet Affairs Med Ctr, Nashville, TN 37212 USA. NIMH, Mol Biol Lab, Bethesda, MD 20892 USA. RP Stabler, SP (reprint author), Univ Colorado, Hlth Sci Ctr, Dept Med, Campus Box B-170,4200 E 9th Ave, Denver, CO 80262 USA. RI Wahl, Markus/D-6365-2017 OI Wahl, Markus/0000-0002-2811-5307 FU NIA NIH HHS [AG-09834] NR 52 TC 44 Z9 44 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0026-0495 J9 METABOLISM JI Metab.-Clin. Exp. PD AUG PY 2002 VL 51 IS 8 BP 981 EP 988 DI 10.1053/meta.2002.34017 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 578UF UT WOS:000177138000007 PM 12145770 ER PT J AU Lee, Y Kim, HK Park, HE Park, MH Joe, YA AF Lee, Y Kim, HK Park, HE Park, MH Joe, YA TI Effect of N-1-guanyl-1,7-diaminoheptane, an inhibitor of deoxyhypusine synthase, on endothelial cell growth, differentiation and apoptosis SO MOLECULAR AND CELLULAR BIOCHEMISTRY LA English DT Article DE N-1-guanyl-1,7-diaminoheptane; hypusine; eIF5A; endothelial cells; proliferation ID YEAST SACCHAROMYCES-CEREVISIAE; INITIATION-FACTOR 5A; HAMSTER OVARY CELLS; PROTEIN-SYNTHESIS; HYPUSINE FORMATION; FACTOR EIF-4D; AMINO-ACID; SPERMIDINE; IDENTIFICATION; PURIFICATION AB An unusual amino acid, hypusine [N-epsilon-(4-amino-2-hydroxybutyl)lysine], is formed post-translationally in a single cellular protein, the eukaryotic translation initiation factor 5A (eIF5A) by deoxyhypusine synthase and deoxyhypusine hydroxylase. Although eIF5A and its hypusine modification are essential for eukaryotic cell viability, the true physiological function of eIF5A is yet unknown. We have examined the effects of N-1-guanyl-1,7-diaminoheptane (GC7), a potent inhibitor of deoxyhypusine synthase, on endothelial cell proliferation, differentiation and apoptosis. Upon treatment of human umbilical vein endothelial cells (HUVEC) with GC7, dose-dependent inhibition of hypusine formation and cellular proliferation was observed. GC7 at 10 muM caused almost complete inhibition of cellular hypusine synthesis and led to cytostasis of HUVEC. Pretreatment of HUVEC with GC7 up to 50 muM for 4 days had little effect on the attachment and differentiation of these cells on Matri-gel and did not cause induction of apoptosis. Instead, the GC7 pretreatment (96 h at 5-50 muM) elicited protective effects against apoptotic death of HUVEC induced by serum starvation. These results suggest that eIF-5A may be involved in expression of proteins essential for apoptosis of endothelial cells as well as those for cellular proliferation. C1 Catholic Univ Korea, Catholic Res Inst Med Sci, Canc Res Inst, Seoul 137701, South Korea. NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD USA. RP Joe, YA (reprint author), Catholic Univ Korea, Catholic Res Inst Med Sci, Canc Res Inst, Banpodong 505, Seoul 137701, South Korea. NR 36 TC 30 Z9 35 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0300-8177 J9 MOL CELL BIOCHEM JI Mol. Cell. Biochem. PD AUG PY 2002 VL 237 IS 1-2 BP 69 EP 76 DI 10.1023/A:1016535217038 PG 8 WC Cell Biology SC Cell Biology GA 578VX UT WOS:000177142700008 PM 12236588 ER PT J AU Antunes, A Templeton, AR Guyomard, R Alexandrino, P AF Antunes, A Templeton, AR Guyomard, R Alexandrino, P TI The role of nuclear genes in intraspecific evolutionary inference: Genealogy of the transferrin gene in the brown trout SO MOLECULAR BIOLOGY AND EVOLUTION LA English DT Article DE brown trout; gene conversion; nuclear genealogy; recombination; Salmo trutta; salmonids; transferrin ID SALMO-TRUTTA L.; LIPOPROTEIN-LIPASE GENE; DNA-SEQUENCE VARIATION; MITOCHONDRIAL-DNA; PHYLOGENETIC-RELATIONSHIPS; PHENOTYPIC ASSOCIATIONS; ONCORHYNCHUS-KISUTCH; CLADOGRAM ESTIMATION; POPULATION-STRUCTURE; CLADISTIC-ANALYSIS AB Technical and biological hurdles have precluded the retrieval of nuclear gene genealogies within most species. Among these obstacles, the possibility of intragenic recombination is one of the most demanding challenges. We studied the utility of nuclear genes for intraspecific evolutionary inferences by selecting a nuclear gene that exhibits patterns of considerable geographic differentiation in the brown trout (Salmo trutta) species complex. Haplotype variation from a nucleotide sequence of similar to3.7 kb encompassing a portion of the transferrin (TF) gene was surveyed in 31 brown trout individuals collected across the native Eurasian range. Statistically significant recombination and gene conversion events were detected. However, we showed that the substantial cladistic structure was not disrupted by recombination or gene conversion events and the additional structure was estimated to have emerged after those events. Because loci with unusually high levels of variation might indicate the presence of selection, we tested the hypothesis of neutrality and found some evidence for directional selection. The strong geographic signal observed in the TF genealogy, coupled with the current spatial distribution of electromorphs, gave us the ability to draw empirical phylogeographic inferences. We delineated the composition of current brown trout populations on the basis of 3,625 individuals electrophoretically scored for the TF locus. We hypothesized scenarios of historical radiation and dispersal events, thus providing new insights refining previous allozyme and mtDNA inferences. We infer that the most ancestral brown trout populations inhabited tributaries from the Black, Caspian, and Aral Sea drainages. An early radiation of the species occurred throughout the Mediterranean, followed by independent dispersal events from the Adriatic to the southernmost Iberian Atlantic and, more recently, a rapid expansion throughout most of the Atlantic drainages. C1 CIBIO UP, Ctr Invest Biodivers & Recursos Genet, Vairao, Portugal. Univ Porto, Fac Ciencias, Dept Zool & Anthropol, Oporto, Portugal. Washington Univ, Dept Biol, St Louis, MO 63130 USA. INRA, Lab Genet Poissons, Jouy En Josas, France. RP Antunes, A (reprint author), NCI, Lab Genom Divers, Frederick, MD 21702 USA. EM aantunes@hotmail.com RI Alexandrino, Paulo/A-3582-2010; Templeton, Alan/F-5963-2011; OI Alexandrino, Paulo/0000-0002-8968-2452; Antunes, Agostinho/0000-0002-1328-1732 NR 80 TC 41 Z9 43 U1 1 U2 8 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0737-4038 EI 1537-1719 J9 MOL BIOL EVOL JI Mol. Biol. Evol. PD AUG PY 2002 VL 19 IS 8 BP 1272 EP 1287 PG 16 WC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity GA 587MB UT WOS:000177643900008 PM 12140239 ER PT J AU Thakurta, AG Whalen, WA Yoon, JH Bharathi, A Kozak, L Whiteford, C Love, DC Hanover, JA Dhar, R AF Thakurta, AG Whalen, WA Yoon, JH Bharathi, A Kozak, L Whiteford, C Love, DC Hanover, JA Dhar, R TI Crp79p, like Mex67p, is an auxiliary mRNA export factor in Schizosaccharomyces pombe SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID MESSENGER-RNA EXPORT; NUCLEAR-PORE COMPLEX; EXON-EXON JUNCTIONS; FISSION YEAST; FUNCTIONALLY INTERACTS; BINDING-PROTEIN; POLY(A)(+) RNA; HNRNP A1; TRANSPORT; NUCLEOPORIN AB The export of mRNA from the nucleus to the cytoplasm involves interactions of proteins with mRNA and the nuclear pore complex. We isolated Crp79p, a novel mRNA export factor from the same synthetic lethal screen that led to the identification of spMex67p in Schizosaccharomyces pombe. Crp79p is a 710-amino-acid-long protein that contains three RNA recognition motif domains in tandem and a distinct C-terminus. Fused to green fluorescent protein (GFP), Crp79p localizes to the cytoplasm. Like Mex67p, Crp79-GFP binds poly(A)(+) RNA in vivo, shuttles between the nucleus and the cytoplasm, and contains a nuclear export activity at the C-terminus that is Crm1p-independent. All of these properties are essential for Crp79p to promote mRNA export. Crp79p import into the nucleus depends on the Ran system. A domain of spMex67p previously identified as having a nuclear export activity can functionally substitute for the nuclear export activity at the C-terminus of Crp79p. Although both Crp79p and spMex67p function to export mRNA, Crp79p does not substitute for all of spMex67p functions and probably is not a functional homologue of spMex67p. We propose that Crp79p is a nonessential mRNA export carrier in S. pombe. C1 NCI, Basic Res Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Lab Cell Biochem & Biol, NIH, Bethesda, MD 20892 USA. RP NCI, Basic Res Lab, NIH, Bethesda, MD 20892 USA. EM dharr@mail.nih.gov NR 51 TC 8 Z9 9 U1 0 U2 1 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 EI 1939-4586 J9 MOL BIOL CELL JI Mol. Biol. Cell PD AUG PY 2002 VL 13 IS 8 BP 2571 EP 2584 DI 10.1091/mbc.E01.11.0133 PG 14 WC Cell Biology SC Cell Biology GA 583PM UT WOS:000177416300004 PM 12181330 ER PT J AU Horikawa, I Cable, PL Mazur, SJ Appella, E Afshari, CA Barrett, JC AF Horikawa, I Cable, PL Mazur, SJ Appella, E Afshari, CA Barrett, JC TI Downstream E-box-mediated regulation of the human telomerase reverse transcriptase (hTERT) gene transcription: Evidence for an endogenous mechanism of transcriptional repression SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID CATALYTIC SUBUNIT GENE; NORMAL HUMAN-CELLS; C-MYC; CANCER-CELLS; DNA-BINDING; PROMOTER REGION; TUMOR-CELLS; IN-VIVO; EXPRESSION; IDENTIFICATION AB Regulation of the hTERT gene encoding the telomerase catalytic subunit plays an important role in human cell senescence, immortalization, and carcinogenesis. By examining the activity of various deleted or mutated hTERT promoter fragments, we show that an E-box element downstream of the transcription initiation site is critical to differential hTERT transcription between the telomerase/hTERT-positive renal cell carcinoma cell line (RCC23) and its telomerase/hTERT-negative counterpart containing a transferred, normal chromosome 3 (RCC23+3). This E-box element mediated repression of hTERT transcription in RCC23+3 but not in RCC23. A copy number-dependent enhancement of the repression suggested active repression, rather than loss of activation, in RCC23+3. Endogenous expression levels of c-Myc or Mad1, which could activate or repress hTERT transcription When overexpressed, did not account for the differential hTERT transcription. Gel mobility shift assays identified the upstream stimulatory factors (USFs) as a major E-box-binding protein complex in both RCC23 and RCC23+3 and, importantly, detected an RCC23+3-specific, E-box-binding factor that was distinct from the USF and Myc/Mad families. The E-box-mediated repression was also active in normal human fibroblasts and epithelial cells and inactive in some, but not all, telomerase/hTERT-positive cancer cells. These findings provide evidence for an endogenous, repressive mechanism that actively functions in telomerase/hTERT-negative normal cells and becomes defective during carcinogenic processes, e.g., by an inactivation of the telomerase repressor gene on chromosome 3. C1 NCI, Lab Biosyst & Canc, Canc & Aging Sect, NIH, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, Chem Immunol Sect, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Horikawa, I (reprint author), NCI, Lab Biosyst & Canc, Canc & Aging Sect, NIH, Bethesda, MD 20892 USA. NR 51 TC 72 Z9 89 U1 1 U2 5 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD AUG PY 2002 VL 13 IS 8 BP 2585 EP 2597 DI 10.1091/mbc.E01.11.0107 PG 13 WC Cell Biology SC Cell Biology GA 583PM UT WOS:000177416300005 PM 12181331 ER PT J AU Roh, DH Bowers, B Schmidt, M Cabib, E AF Roh, DH Bowers, B Schmidt, M Cabib, E TI The septation apparatus, an autonomous system in budding yeast SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID SACCHAROMYCES-CEREVISIAE; CELL-WALL; CYTOKINESIS; PROTEIN; CHITIN; RING; ACTOMYOSIN; MORPHOGENESIS; DYNAMICS; SEPTUM AB Actomyosin ring contraction and chitin primary septum deposition are interdependent processes in cell division of budding yeast. By fusing Myo1p, as representative of the contractile ring, and Chs2p for the primary septum, to different fluorescent proteins we show herein that the two processes proceed essentially at the same location and simultaneously. Chs2p differs from Myo1p in that it reflects the changes in shape of the plasma membrane to which it is attached and in that it is packed after its action into visible endocytic vesicles for its disposal. To ascertain whether this highly coordinated system could function independently of other cell cycle events, we reexamined the septum-like structures made by the septin mutant cdc3 at various sites on the cell cortex at the nonpermissive temperature. With the fluorescent fusion proteins mentioned above, we observed that in cdc3 at 37degreesC both Myo1p and Chs2p colocalize at different spots of the cell cortex. A contraction of the Myo1p patch could also be detected, as well as that of a Chs2p patch, with subsequent appearance of vesicles. Furthermore, the septin Cdc12p, fused with yellow or cyan fluorescent protein, also colocalized with Myo1p and Chs2p at the aberrant locations. The formation of delocalized septa did not require nuclear division. We conclude that the septation apparatus, composed of septins, contractile ring, and the chitin synthase II system, can function at ectopic locations autonomously and independently of cell division, and that it can recruit the other elements necessary for the formation of secondary septa. C1 NIDDKD, Lab Biochem & Genet, Bethesda, MD 20892 USA. NHLBI, Cell Biol Lab, Bethesda, MD 20892 USA. RP Cabib, E (reprint author), NIDDKD, Lab Biochem & Genet, Bethesda, MD 20892 USA. NR 21 TC 50 Z9 51 U1 0 U2 0 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD AUG PY 2002 VL 13 IS 8 BP 2747 EP 2759 DI 10.1091/mbc.E02-03-0158 PG 13 WC Cell Biology SC Cell Biology GA 583PM UT WOS:000177416300017 PM 12181343 ER PT J AU Huang, Y Xu, L Sun, YN Nabel, GJ AF Huang, Y Xu, L Sun, YN Nabel, GJ TI The assembly of Ebola virus nucleocapsid requires virion-associated proteins 35 and 24 and posttranslational modification of nucleoprotein SO MOLECULAR CELL LA English DT Article ID LINKED N-ACETYLGLUCOSAMINE; MARBURG-VIRUS; MAMMALIAN-CELLS; O-GLCNAC; REPLICATION; GLYCOSYLATION; PARTICLES; VP40; GAG; TRANSCRIPTION AB Ebola virus encodes seven viral structural and regulatory proteins that support its high rates of replication, but little is known about nucleocapsid assembly of this virus in infected cells. We report here that three viral proteins are necessary and sufficient for formation of Ebola virus particles and that intracellular posttranslational modification regulates this process. Expression of the nucleoprotein (NP) and virion-associated proteins VP35 and VP24 led to spontaneous assembly of nucleocapsids in transfected 293T cells by transmission electron microscopy. A specific biochemical interaction of these three proteins was demonstrated, and, interestingly, O-glycosylation and sialation of NP were demonstrated and necessary for their association. This distinct mechanism of regulation for filovirus assembly suggests new approaches for viral therapies and vaccines for Ebola and related viruses. C1 NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. RP Nabel, GJ (reprint author), NIAID, Vaccine Res Ctr, NIH, 40 Convent Dr, Bethesda, MD 20892 USA. NR 46 TC 109 Z9 128 U1 2 U2 22 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell PD AUG PY 2002 VL 10 IS 2 BP 307 EP 316 DI 10.1016/S1097-2765(02)00588-9 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 586LD UT WOS:000177584700014 PM 12191476 ER PT J AU Hickman, AB Ronning, DR Kotin, RM Dyda, F AF Hickman, AB Ronning, DR Kotin, RM Dyda, F TI Structural unity among viral origin binding proteins: Crystal structure of the nuclease domain of adeno-associated virus Rep SO MOLECULAR CELL LA English DT Article ID ROLLING-CIRCLE REPLICATION; DNA-REPLICATION; ESCHERICHIA-COLI; TOPOISOMERASE-I; SITE; GENE; RNA; INTEGRATION; INITIATION; AAV AB Adeno-associated virus (AAV), unique among animal viruses in itsability to integrate into a specific chromosomal location, is a promising vector for human gene therapy. AAV Replication (Rep) protein is essential for viral replication and integration, and its amino terminal domain possesses site-specific DNA binding and endonuclease activities required for replication initiation and integration. This domain displays a novel endonuclease fold and demonstrates an unexpected structural relationship to other viral origin binding proteins such as the papillomavirus E1 protein and the SV40 T antigen. The active site, located at the bottom of a positively charged cleft, is formed by the spatial convergence of a divalent metal ion and two conserved sequence motifs that define the rolling circle replication superfamily. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. RP Hickman, AB (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. EM ahickman@helix.nih.gov RI kotin, robert/B-8954-2008 NR 61 TC 87 Z9 90 U1 0 U2 5 PU CELL PRESS PI CAMBRIDGE PA 600 TECHNOLOGY SQUARE, 5TH FLOOR, CAMBRIDGE, MA 02139 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell PD AUG PY 2002 VL 10 IS 2 BP 327 EP 337 DI 10.1016/S1097-2765(02)00592-0 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 586LD UT WOS:000177584700016 PM 12191478 ER PT J AU Morrison, RS Kinoshita, Y Johnson, MD Uo, T Ho, JT McBee, JK Conrads, TP Veenstra, TD AF Morrison, RS Kinoshita, Y Johnson, MD Uo, T Ho, JT McBee, JK Conrads, TP Veenstra, TD TI Proteomic analysis in the neurosciences SO MOLECULAR & CELLULAR PROTEOMICS LA English DT Review ID GLYCOGEN-SYNTHASE KINASE-3; CODED AFFINITY TAGS; 2-DIMENSIONAL GEL-ELECTROPHORESIS; INTERACTING PROTEIN KINASE-2; TAU TRANSGENIC MICE; MASS-SPECTROMETRY; BRAIN PROTEINS; ALZHEIMERS-DISEASE; MESSENGER-RNA; KAINIC ACID AB Proteomics is a field of study directed toward providing a comprehensive view of the characteristics and activity of every cellular protein. Rapid innovations in the core technologies required to characterize proteins on a global scale are poised to bring about a comprehensive understanding of how dynamic changes in protein expression, post-translational modification, and function affect complex signaling and regulatory networks. These advances have significant implications for understanding the multitude of pathways that govern behavior and cognition and the response of the nervous system to injury and disease. C1 Univ Washington, Sch Med, Dept Neurol Surg, Seattle, WA 98195 USA. Univ Washington, Sch Med, Dept Ophthalmol, Seattle, WA 98195 USA. NCI, SAIC Frederick Inc, NIH, Frederick, MD 21702 USA. RP Morrison, RS (reprint author), Univ Washington, Sch Med, Dept Neurol Surg, Seattle, WA 98195 USA. FU NEI NIH HHS [EY07031]; NINDS NIH HHS [NS35533, NS42123, NS42699]; PHS HHS [N01-C0-12400] NR 54 TC 44 Z9 46 U1 0 U2 9 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 1535-9476 J9 MOL CELL PROTEOMICS JI Mol. Cell. Proteomics PD AUG PY 2002 VL 1 IS 8 BP 553 EP 560 DI 10.1074/mcp.R200004-MCP200 PG 8 WC Biochemical Research Methods SC Biochemistry & Molecular Biology GA 654VA UT WOS:000181515700001 PM 12376570 ER PT J AU Yudt, MR Cidlowski, JA AF Yudt, MR Cidlowski, JA TI The glucocorticoid receptor: Coding a diversity of proteins and responses through a single gene SO MOLECULAR ENDOCRINOLOGY LA English DT Review ID LIGAND-BINDING DOMAIN; MOUSE LYMPHOMA-CELLS; BETA-ISOFORM; DNA-BINDING; CORTISOL RESISTANCE; MESSENGER-RNA; TRANSCRIPTION FACTOR; MOLECULAR-BASIS; POINT MUTATION; LEUKEMIC-CELLS AB The ability of natural and synthetic glucocorticoids to elicit numerous and diverse physiological responses is remarkable. How the product of a single gene can participate in such a myriad of cell- and tissue-specific pathways has remained largely unknown. The last several years have seen increased description of glucocorticoid receptor (GR) protein isoforms. Here we review the current state of knowledge regarding naturally occurring GR isoforms and discuss how this array of receptor species generates the diversity associated with the glucocorticoid response. We propose that the multiplicity of receptor forms have unique tissue-specific actions on the downstream biology providing a mechanism to create GR signaling networks. C1 Natl Inst Environm Hlth Sci, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Cidlowski, JA (reprint author), Natl Inst Environm Hlth Sci, Lab Signal Transduct, NIH, 111 TW Alexander Dr,NC MDF307, Res Triangle Pk, NC 27709 USA. NR 80 TC 159 Z9 167 U1 3 U2 11 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD AUG PY 2002 VL 16 IS 8 BP 1719 EP 1726 DI 10.1210/me.2002-0106 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 577WV UT WOS:000177086400001 PM 12145329 ER PT J AU Prufer, K Barsony, J AF Prufer, K Barsony, J TI Retinoid X receptor dominates the nuclear import and export of the unliganded vitamin D receptor SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID THYROID-HORMONE RECEPTOR; DNA-BINDING; PROGESTERONE-RECEPTOR; GLUCOCORTICOID RECEPTOR; LOCALIZATION SIGNAL; TARGETED ABLATION; PROTEIN IMPORT; ZINC FINGERS; TRANSCRIPTION; TRAFFICKING AB Liganded and unliganded vitamin D receptors (VDRs) carry out distinct functions; both types of functions require heterodimerization with retinoid X receptors (RXRs). Our recent studies with fluorescent protein chimeras of VDR and RXR, termed GFP-VDR, YFP-RXR, and RXR-BFP, indicated that RXR regulates VDR functions in part by regulating subcellular localization. Here we explored the mechanisms of this regulation. Photobleaching experiments demonstrated that YFP-RXR and both unliganded and liganded GFP-VDR shuttle constantly between nucleus and cytoplasm. To characterize RXR import, we identified a nuclear localization sequence (NLS) in the DNA-binding domain. Mutations in this NLS caused predominant cytoplasmic localization of n/sYFP-RXR and prevented transcriptional activity. The n/sRXR-BFP retained unliganded GFP-VDR in the cytoplasm and reduced baseline transcriptional activity. After calcitriol exposure, however, both GFP-VDR and n/sRXR-BFP entered the nucleus. We characterized receptor export rates and mechanisms using permeabilization experiments. Mutations in the calreticulin binding region slowed both GFP-VDR and YFP-RXR export. Coexpression of RXR-BFP slowed the export of unliganded GFP-VDR, whereas calcitriol treatment tripled the rate of GFP-VDR export. Treatment with leptomycin B, an inhibitor of CRM-1 receptor-mediated export, inhibited export of unliganded GFP-VDR but did not influence export of liganded GFP-VDR or YFP-RXR. Leptomycin B added before calcitriol similarly decreased hormone-induced luciferase activity but was ineffective when added subsequent to calcitriol. These results indicate that the unliganded and liganded VDR interact differently with the import and export receptors and with RXR. Most likely, the regulation of VDR nuclear import by RXR is essential for ligand-independent functions. C1 NIDDK, LCBB, NIH, Bethesda, MD 20892 USA. RP Barsony, J (reprint author), NIDDK, LCBB, NIH, 8 Ctr Dr,Room 422, Bethesda, MD 20892 USA. NR 49 TC 92 Z9 94 U1 2 U2 8 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD AUG PY 2002 VL 16 IS 8 BP 1738 EP 1751 DI 10.1210/me.2001-0345 PG 14 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 577WV UT WOS:000177086400003 PM 12145331 ER PT J AU Bastepe, M Gunes, Y Perez-Villamil, B Hunzelman, J Weinstein, LS Juppner, H AF Bastepe, M Gunes, Y Perez-Villamil, B Hunzelman, J Weinstein, LS Juppner, H TI Receptor-mediated adenylyl cyclase activation through XL alpha s, the extra-large variant of the stimulatory G protein alpha-subunit SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID MOUSE GNAS LOCUS; HEREDITARY OSTEODYSTROPHY; PARATHYROID-HORMONE; GENE; EXPRESSION; TRANSCRIPTS; ANTISENSE; PSEUDOHYPOPARATHYROIDISM; METHYLATION; RESIDUES AB XLalphas, the large variant of the stimulatory G protein a subunit (Gsalpha), is derived from GNAS1 through the use of an alternative first exon and promoter. Gsa and XLalphas have distinct amino-terminal domains, but are identical over the carboxyl-terminal portion encoded by exons 2-13. XLalphas can mimic some functions of Gsalpha, including betagamma interaction and adenylyl cyclase stimulation. However, previous attempts to demonstrate coupling of XLalphas to typically Gs-coupled receptors have not been successful. We now report the generation of murine cell lines that carry homozygous disruption of Gnas exon 2, and are therefore null for endogenous XLalphas and Gsalpha (Gnas(E2-/E2-)). Gnas(E2-/E2-) cells transfected with plasmids encoding XLalphas and different heptahelical receptors, including the beta2-adrenergic receptor and receptors for PTH, TSH, and CRF, showed agonist-mediated cAMP accumulation that was indistinguishable from that observed with cells transiently coexpressing Gsalpha and these receptors. Our findings thus indicate that XLalphas is capable of functionally coupling to receptors that normally act via Gsalpha. C1 Massachusetts Gen Hosp, Endocrine Unit, Boston, MA 02114 USA. Massachusetts Gen Hosp, Ctr Canc, Boston, MA 02114 USA. MassGen Hosp Children, Boston, MA 02114 USA. Harvard Univ, Sch Med, Boston, MA 02114 USA. NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Juppner, H (reprint author), Massachusetts Gen Hosp, Endocrine Unit, 50 Blossom St,WEL 5, Boston, MA 02114 USA. FU PHS HHS [R01 46718-06] NR 40 TC 86 Z9 87 U1 2 U2 3 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD AUG PY 2002 VL 16 IS 8 BP 1912 EP 1919 DI 10.1210/me.2002-0054 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 577WV UT WOS:000177086400016 PM 12145344 ER PT J AU Montagnani, M Ravichandran, LV Chen, H Esposito, DL Quon, MJ AF Montagnani, M Ravichandran, LV Chen, H Esposito, DL Quon, MJ TI Insulin receptor substrate-1, and phosphoinositide-dependent kinase-1 are required for insulin-stimulated production of nitric oxide in endothelial SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID RAT ADIPOSE-CELLS; PROTEIN-KINASE; PHOSPHATIDYLINOSITOL 3-KINASE; C-ZETA; GLUCOSE-TRANSPORTER-4 TRANSLOCATION; GLUT4 TRANSLOCATION; GLUCOSE-TRANSPORT; FULL ACTIVATION; PHOSPHORYLATION; CALMODULIN AB Vasodilator actions of insulin are mediated by signaling pathways involving phosphatidylinositol 3-kinase (PI 3-kinase) and Akt that lead to activation of endothelial nitric oxide synthase (eNOS) in endothelium. Signaling molecules immediately upstream and downstream from PI 3-kinase involved with production of NO in response to insulin have not been previously identified. In this study, we evaluated roles of insulin receptor substrate 1 (IRS-1) and phosphoinositide-dependent kinase 1 (PDK-1) in production of NO. The fluorescent dye 4,5-diamine fluorescein diacetate was used to directly measure NO in NIH-3T3(IR) cells transiently cotransfected with eNOS and various IRS-1 or PDK-1 constructs. In control cells, transfected with only eNOS, insulin stimulated a rapid dose-dependent increase in NO. Overexpression of wild-type IRS-1 increased the maximal insulin response 3-fold. Overexpression of IRS1-F6 (mutant that does not bind PI 3-kinase) or an antisense ribozyme against IRS-1 substantially inhibited insulin-stimulated production of NO. Likewise, overexpression of wild-type PDK-1 enhanced insulin-stimulated production of NO, whereas a kinase-inactive mutant PDK-1 inhibited this action of insulin. Qualitatively similar results were observed in vascular endothelial cells. Production of NO by a calcium-dependent mechanism in response to lysophosphatidic acid was unaffected by either wild-type or mutant IRS-1 and PDK-1. We conclude that IRS-1 and PDK-1 play necessary roles in insulin-signaling pathways leading to activation of eNOS. Furthermore, classical Ca2+-mediated pathways for activation of eNOS are separable from IRS-1- and PDK-1-dependent insulin-signaling pathways. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. Univ G Annunzio, Dept Oncol & Neurosci, I-66013 Chieti, Italy. RP Quon, MJ (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,Room 8C-218,10 Ctr Dr MSC 1755, Bethesda, MD 20892 USA. RI Quon, Michael/B-1970-2008; OI montagnani, monica/0000-0002-5697-8185 NR 50 TC 141 Z9 151 U1 0 U2 1 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD AUG PY 2002 VL 16 IS 8 BP 1931 EP 1942 DI 10.1210/me.2002-0074 PG 12 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 577WV UT WOS:000177086400018 PM 12145346 ER PT J AU McDonald, JD Andriolo, M Cali, F Mirisola, M Puglisi-Allegra, S Romano, V Sarkissian, CN Smith, CB AF McDonald, JD Andriolo, M Cali, F Mirisola, M Puglisi-Allegra, S Romano, V Sarkissian, CN Smith, CB TI The phenylketonuria mouse model: a meeting review SO MOLECULAR GENETICS AND METABOLISM LA English DT Review ID PHENYLALANINE-HYDROXYLASE; MATERNAL PHENYLKETONURIA C1 Wichita State Univ, Dept Biol Sci, Wichita, KS 67208 USA. OASI Ist Ric Ritardo Mentale & Involuz Cerebrale, Troina, Sicily, Italy. Univ Palermo, Dept Biopathol & Biomed Methodol, Palermo, Sicily, Italy. Univ Rome, Dept Psychol, Rome, Italy. Fdn Santa Lucia IRCCS, Rome, Italy. McGill Univ, Montreal Childrens Hosp, Res Inst, Montreal, PQ H3H 1P3, Canada. NIMH, Cerebral Metab Lab, Bethesda, MD 20892 USA. RP McDonald, JD (reprint author), Wichita State Univ, Dept Biol Sci, Wichita, KS 67208 USA. RI Cali, Francesco/D-9178-2017; OI Cali, Francesco/0000-0001-8667-8223; Romano, Valentino/0000-0003-3238-5702; Scavuzzo, Cataldo/0000-0002-3905-4191 FU Telethon [E.1259] NR 23 TC 16 Z9 16 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1096-7192 J9 MOL GENET METAB JI Mol. Genet. Metab. PD AUG PY 2002 VL 76 IS 4 BP 256 EP 261 AR PII S1096-7192(02)00115-4 PG 6 WC Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental SC Endocrinology & Metabolism; Genetics & Heredity; Research & Experimental Medicine GA 593WV UT WOS:000178018200003 PM 12208130 ER PT J AU Orvisky, E Park, JK LaMarca, ME Ginns, EI Martin, BM Tayebi, N Sidransky, E AF Orvisky, E Park, JK LaMarca, ME Ginns, EI Martin, BM Tayebi, N Sidransky, E TI Glucosylsphingosine accumulation in tissues from patients with Gaucher disease: correlation with phenotype and genotype SO MOLECULAR GENETICS AND METABOLISM LA English DT Article DE Gaucher disease; glucocerebrosidase; glucosylsphingosine; lysosomal storage; high performance liquid chromatography ID PROTEIN-KINASE-C; LYSOSULFATIDE SULFOGALACTOSYLSPHINGOSINE; METACHROMATIC LEUKODYSTROPHY; GLUCOCEREBROSIDASE GENE; PSYCHOSINE; LYSOSPHINGOLIPIDS; GLUCOSYLCERAMIDE; DIFFERENTIATION; SPHINGOSINE; MUTATIONS AB Gaucher disease, the inherited deficiency of lysosomal glucocerebrosidase, presents with a wide spectrum of clinical manifestations including neuronopathic and non-neuronopathic forms. While the lipid glucosylceramide is stored in both patients with Gaucher disease and in a null allele mouse model of Gaucher disease, elevated levels of a second potentially toxic substrate, glucosylsphingosine, are also found. Using high performance liquid chromatography, glucosylsphingosine levels were measured in tissues from patients with type 1, 2, and 3 Gaucher disease. Glucosylsphingosine was measured in 16 spleen samples (8 type 1; 4 type 2; and 4, type 3) and levels ranged from 54 to 728 ng/mg protein in the patients with type I disease, 133 to 1200 ng/mg protein in the patients with type 2, and 109 to 1298 ng/mg protein in the type 3 samples. The levels of splenic glucosylsphingosine bore no relation to the type of Gaucher disease, the age of the patient, the genotype, nor the clinical course. In the same patients, hepatic glucosylsphingosine levels were lower than in spleen. Glucosylsphingosine was also measured in brains from 13 patients (I type 1; 8 type 2; and 4 type 3). While the glucosylsphingosine level in the brain from the type 1 patient, 1.0 ng/mg protein, was in the normal range, the levels in the type 3 samples ranged from 14 to 32 ng/mg protein, and in the type 2 samples from 24 to 437 ng/mg protein, with the highest values detected in two fetuses with hydrops fetalis. The elevated levels found in brains from patients with neuronopathic Gaucher disease support the hypothesis that glucosylsphingosine may contribute to the nervous system involvement in these patients. (C) 2002 Elsevier Science (USA). All rights reserved. C1 NIMH, Clin Neurosci Branch, NIH, Bethesda, MD 20892 USA. RP Sidransky, E (reprint author), NIMH, Clin Neurosci Branch, NIH, Bethesda, MD 20892 USA. NR 37 TC 79 Z9 81 U1 1 U2 6 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1096-7192 J9 MOL GENET METAB JI Mol. Genet. Metab. PD AUG PY 2002 VL 76 IS 4 BP 262 EP 270 AR PII S1096-7192(02)00117-8 DI 10.1016/S1096-7192(02)00117-8 PG 9 WC Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental SC Endocrinology & Metabolism; Genetics & Heredity; Research & Experimental Medicine GA 593WV UT WOS:000178018200004 PM 12208131 ER PT J AU Nash, TE AF Nash, TE TI Surface antigenic variation in Giardia lamblia SO MOLECULAR MICROBIOLOGY LA English DT Review ID EXPERIMENTAL HUMAN INFECTIONS; GENE-EXPRESSION; ZINC-BINDING; PROTEIN VSP; MICE; ANTIBODIES; EUKARYOTE; DNA; IDENTIFICATION; VARIABILITY AB Giardia lamblia, a common intestinal dwelling protozoan and a cause of diarrhoea in humans and animals world-wide, undergoes surface antigenic variation. The variant-specific surface proteins (VSPs) are a family of related, highly unusual proteins that cover the entire surface of the parasite. VSPs are cysteine-rich proteins containing many CXXC motifs, one or two GGCY motifs, a conserved hydrophobic tail and a Zn finger motif. The biological role(s) of VSPs is unclear. As VSPs are resistant to the effects of intestinal proteases, they likely allow the organism to survive in the protease-rich small intestine. Although immune escape is commonly mentioned as the reason antigenic variation occurs, VSP expression changes in vivo even in the absence of an adaptive immune system suggesting the biological role of antigenic variation is more complex. The molecular mechanisms involved in antigenic variation are not known but appear to differ from those known to occur in other protozoa. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Nash, TE (reprint author), NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. NR 41 TC 84 Z9 86 U1 0 U2 2 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD AUG PY 2002 VL 45 IS 3 BP 585 EP 590 DI 10.1046/j.1365-2958.2002.03029.x PG 6 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 577XF UT WOS:000177087600001 PM 12139606 ER PT J AU Fu, LZ Buchholz, D Shi, YB AF Fu, LZ Buchholz, D Shi, YB TI Novel double promoter approach for identification of transgenic animals: A tool for in vivo analysis of gene function and development of gene-based therapies SO MOLECULAR REPRODUCTION AND DEVELOPMENT LA English DT Article DE transgenesis; green fluorescent protein; Xenopus laevis; matrix metalloproteinase ID GREEN FLUORESCENT PROTEIN; HORMONE RESPONSE GENES; EXPRESSION PATTERN; XENOPUS EMBRYOS; STROMELYSIN-3; APOPTOSIS; SELECTION; PROGRAMS; LAEVIS; MARKER AB Advances in vertebrate genetics have allowed studies of gene function in developing animals through gene knockout and transgenic analyses. These advances have encouraged the development of gene-based therapies through introduction of exogenous genes to enhance and/or replace dysfunctional or missing genes. However, in vertebrates, such analyses often involve tedious screening for transgenic animals, such as PCR-based genotype determinations. Here, we report the use of double-promoter plasmids carrying the transgene of interest and the crystallin-promotor-driven Green fluorescent protein (GFP) in transgenic Xenopus laevis tadpoles. This strategy allows a simple examination for the presence of GFP in the eyes to identify transgenic animals. PCR-based genotyping and functional characterization confirms that all animals expressing GFP in the eyes indeed carry the desired promoter/transgene units. Thus, the use of this and other similar vectors should dramatically improve current transgenesis protocols and reduce the time and cost for identifying transgenic animals. (C) 2002 Wiley-Liss, Inc. C1 NICHD, Lab Gene Regulat & Dev, UNit Mol Morphogenesis, NIH, Bethesda, MD 20892 USA. RP Shi, YB (reprint author), NICHD, Lab Gene Regulat & Dev, UNit Mol Morphogenesis, NIH, Bldg 18T,Rm 106, Bethesda, MD 20892 USA. NR 15 TC 45 Z9 45 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1040-452X J9 MOL REPROD DEV JI Mol. Reprod. Dev. PD AUG PY 2002 VL 62 IS 4 BP 470 EP 476 DI 10.1002/mrd.10137 PG 7 WC Biochemistry & Molecular Biology; Cell Biology; Developmental Biology; Reproductive Biology SC Biochemistry & Molecular Biology; Cell Biology; Developmental Biology; Reproductive Biology GA 571CP UT WOS:000176697500005 PM 12112579 ER PT J AU Caplen, NJ Zheng, ZL Falgout, B Morgan, RA AF Caplen, NJ Zheng, ZL Falgout, B Morgan, RA TI Inhibition of viral gene expression and replication in mosquito cells by dsRNA-triggered RNA interference SO MOLECULAR THERAPY LA English DT Article DE RNA interference; RNAi; mosquito; dsRNA; Semliki Forest virus; dengue virus; gene silencing ID DOUBLE-STRANDED-RNA; MESSENGER-RNA; CAENORHABDITIS-ELEGANS; VIRUS TRANSMISSION; DROSOPHILA CELLS; AEDES-AEGYPTI; C-ELEGANS; HELICASE; DENGUE; DEGRADATION AB Mosquitoes transmit numerous viral pathogens to humans including dengue virus which affects similar to 50 million individuals per year. Inhibition of viral gene expression within an insect host could be used to block virus replication and subsequent transmission of the pathogen to humans. A naturally occurring gene silencing mechanism triggered by double-stranded RNA (dsRNA), RNA interference (RNAi), has recently been described in a number of species including Drosophila. To ascertain if dsRNA-triggered RNAi is present in mosquito cells, we used Aedes albopictus C6/36 cells, and to investigate the feasibility of blocking viral gene expression and replication, we used two mosquito-borne viruses, Semliki Forest virus (SFV) and the serotype 1 dengue virus (DEN1). We demonstrate that dsRNA can specifically inhibit transgene expression in C6/36 cells from both plasmid and SFV replicons and can significantly modify the kinetics of DEN1 RNA and virus replication. The inhibition mediated by dsRNA was sequence-specific and either equal or superior to that induced by antisense single-stranded RNA (ssRNA). This study demonstrates dsRNA-triggered inhibition of gene expression and virus replication in mosquito cells and suggests that this mechanism could be used to block pathogen replication within an insect host and, thus, block disease transmission. C1 NHGRI, NIH, Bethesda, MD 20892 USA. US FDA, Lab Vector Borne Viral Dis, Off Vaccine Res & Review, Ctr Biol Evaluat & Res, Bethesda, MD 20892 USA. RP Morgan, RA (reprint author), NHGRI, NIH, Bethesda, MD 20892 USA. RI Caplen, Natasha/H-2768-2016 OI Caplen, Natasha/0000-0002-0001-9460 NR 45 TC 62 Z9 73 U1 0 U2 6 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1525-0016 J9 MOL THER JI Mol. Ther. PD AUG PY 2002 VL 6 IS 2 BP 243 EP 251 DI 10.1006/mthe.2002.0652 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 582TX UT WOS:000177368200017 PM 12161191 ER PT J AU Thorgeirsson, SS Grisham, JW AF Thorgeirsson, SS Grisham, JW TI Molecular pathogenesis of human hepatocellular carcinoma SO NATURE GENETICS LA English DT Article ID HEPATITIS-B VIRUS; COMPARATIVE GENOMIC HYBRIDIZATION; PRIMARY LIVER-CANCER; LASER CAPTURE MICRODISSECTION; DIFFERENTIAL GENE-EXPRESSION; ABERRANT DNA METHYLATION; HEPATOMA-CELL LINES; MICROSATELLITE INSTABILITY; DYSPLASTIC NODULES; CLONAL ANALYSIS AB Hepatocarcinogenesis is a slow process during which genomic changes progressively alter the hepatocellular phenotype to produce cellular intermediates that evolve into hepatocellular carcinoma. During the long pre-neoplastic stage, in which the liver is often the site of chronic hepatitis, cirrhosis, or both, hepatocyte cycling is accelerated by upregulation of mitogenic pathways, in part through epigenetic mechanisms. This leads to the production of monoclonal populations of aberrant and dysplastic hepatocytes that have telomere erosion and telomerase re-expression, sometimes microsatellite instability, and occasionally structural aberrations in genes and chromosomes. Development of dysplastic hepatocytes in foci and nodules and emergence of hepatocellular carcinoma are associated with the accumulation of irreversible structural alterations in genes and chromosomes, but the genomic basis of the malignant phenotype is heterogeneous. The malignant hepatocyte phenotype may be produced by the disruption of a number of genes that function in different regulatory pathways, producing several molecular variants of hepatocellular carcinoma. New strategies should enable these variants to be characterized. C1 NCI, Expt Carcinogenesis Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA. Univ N Carolina, Dept Pathol & Lab Med, Chapel Hill, NC 27599 USA. RP Thorgeirsson, SS (reprint author), NCI, Expt Carcinogenesis Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA. RI jia, xu/A-8386-2016 NR 96 TC 946 Z9 984 U1 7 U2 73 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD AUG PY 2002 VL 31 IS 4 BP 339 EP 346 DI 10.1038/ng0802-339 PG 8 WC Genetics & Heredity SC Genetics & Heredity GA 578XV UT WOS:000177147100006 PM 12149612 ER PT J AU Lipkin, SM Moens, PB Wang, V Lenzi, M Shanmugarajah, D Gilgeous, A Thomas, J Cheng, J Touchman, JW Green, ED Schwartzberg, P Collins, FS Cohen, PE AF Lipkin, SM Moens, PB Wang, V Lenzi, M Shanmugarajah, D Gilgeous, A Thomas, J Cheng, J Touchman, JW Green, ED Schwartzberg, P Collins, FS Cohen, PE TI Meiotic arrest and aneuploidy in MLH3-deficient mice SO NATURE GENETICS LA English DT Article ID DNA MISMATCH REPAIR; CROSSING-OVER; MUTS HOMOLOG; SACCHAROMYCES-CEREVISIAE; CHROMOSOME SYNAPSIS; MEIOSIS; GENE; RECOMBINATION; MLH1; MSH5 AB MutL homolog 3 (Mlh3) is a member of a family of proteins conserved during evolution and having dual roles in DNA mismatch repair and meiosis(1,2). The pathway in eukaryotes consists of the DNA-binding components, which are the homologs of the bacterial MutS protein (MSH 2-6), and the MutL homologs, which bind to the MutS homologs and are essential for the repair process. Three of the six homologs of MutS that function in these processes, Msh2, Msh3 and Msh6, are involved in the mismatch repair of mutations, frameshifts and replication errors(2,3), and two others, Msh4 and Msh5, have specific roles in meiosis(4-10). Of the four MutL homologs, Mlh1, Mlh3, Pms1 and Pms2, three are involved in mismatch repair and at least two, Pms2 and Mlh1, are essential for meiotic progression in both yeast and mice(2,3,11-14). To assess the role of Mlh3 in mammalian meiosis, we have generated and characterized Mlh3(-/-) mice 15. Here we show that Mlh3(-/-) mice are viable but sterile. Mlh3 is required for Mlh1 binding to meiotic chromosomes and localizes to meiotic chromosomes from the mid pachynema stage of prophase I. Mlh3(-/-) spermatocytes reach metaphase before succumbing to apoptosis, but oocytes fail to complete meiosis I after fertilization. Our results show that Mlh3 has an essential and distinct role in mammalian meiosis. C1 Univ Calif Irvine, Dept Med, Div Hematol Oncol, Irvine, CA 92697 USA. NHGRI, Genet & Mol Biol Branch, Bethesda, MD 20892 USA. York Univ, Dept Biol, N York, ON M3J 1P3, Canada. Albert Einstein Coll Med, Coll Med, Dept Mol Genet, Bronx, NY 10467 USA. NHGRI, Genome Technol Branch, Bethesda, MD 20892 USA. NHGRI, Transgen & Knockout Core Facil, Bethesda, MD 20892 USA. RP Cohen, PE (reprint author), Univ Calif Irvine, Dept Med, Div Hematol Oncol, Irvine, CA 92697 USA. NR 29 TC 194 Z9 205 U1 1 U2 6 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD AUG PY 2002 VL 31 IS 4 BP 385 EP 390 DI 10.1038/ng931 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 578XV UT WOS:000177147100013 PM 12091911 ER PT J AU Castillo-Davis, CI Mekhedov, SL Hartl, DL Koonin, EV Kondrashov, FA AF Castillo-Davis, CI Mekhedov, SL Hartl, DL Koonin, EV Kondrashov, FA TI Selection for short introns in highly expressed genes SO NATURE GENETICS LA English DT Article ID GENOME EVOLUTION; CODON BIAS; C-ELEGANS; DROSOPHILA; LENGTH; TRANSCRIPTION; RATES; SIZE; DNA; CAENORHABDITIS AB Transcription is a slow and expensive process: in eukaryotes, approximately 20 nucleotides can be transcribed per second(1,2) at the expense of at least two ATP molecules per nucleotide(3). Thus, at least for highly expressed genes, transcription of long introns, which are particularly common in mammals, is costly. Using data on the expression of genes that encode proteins in Caenorhabditis elegans and Homo sapiens, we show that introns in highly expressed genes are substantially shorter than those in genes that are expressed at low levels. This difference is greater in humans, such that introns are, on average, 14 times shorter in highly expressed genes than in genes with low expression, whereas in C. elegans the difference in intron length is only twofold. In contrast, the density of introns in a gene does not strongly depend on the level of gene expression. Thus, natural selection appears to favor short introns in highly expressed genes to minimize the cost of transcription and other molecular processes, such as splicing. C1 Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. Harvard Univ, Dept Organism & Evolutionary Biol, Cambridge, MA 02138 USA. RP Natl Ctr Biotechnol Informat, NIH, 8600 Rockville Pike, Bethesda, MD 20894 USA. EM fkondras@ncbi.nlm.nih.gov RI Kondrashov, Fyodor Alexeevich/H-6331-2015 OI Kondrashov, Fyodor Alexeevich/0000-0001-8243-4694 NR 30 TC 304 Z9 312 U1 2 U2 14 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 1061-4036 EI 1546-1718 J9 NAT GENET JI Nature Genet. PD AUG PY 2002 VL 31 IS 4 BP 415 EP 418 DI 10.1038/ng940 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 578XV UT WOS:000177147100019 PM 12134150 ER PT J AU Martin, MP Gao, XJ Lee, JH Nelson, GW Detels, R Goedert, JJ Buchbinder, S Hoots, K Vlahov, D Trowsdale, J Wilson, M O'Brien, SJ Carrington, M AF Martin, MP Gao, XJ Lee, JH Nelson, GW Detels, R Goedert, JJ Buchbinder, S Hoots, K Vlahov, D Trowsdale, J Wilson, M O'Brien, SJ Carrington, M TI Epistatic interaction between KIR3DS1 and HLA-B delays the progression to AIDS SO NATURE GENETICS LA English DT Article ID CELL-INHIBITORY RECEPTOR; NATURAL-KILLER-CELLS; IMMUNE-RESPONSES; T-LYMPHOCYTES; GENE FAMILIES; HIV-1; RECOGNITION; MOLECULES; BW4; POLYMORPHISM AB Natural killer (NK) cells provide defense in the early stages of the innate immune response against viral infections by producing cytokines and causing cytotoxicity(1). The killer immunoglobulin-like receptors (KIRs) on NK cells regulate the inhibition and activation of NK-cell responses through recognition of human leukocyte antigen (HLA) class I molecules on target cells(2). KIR and HLA loci are both highly polymorphic, and some HLA class I products bind and trigger cell-surface receptors specified by KIR genes. Here we report that the activating KIR allele KIR3DS1, in combination with HLA-B alleles that encode molecules with isoleucine at position 80 (HLA-B Bw4-80Ile), is associated with delayed progression to AIDS in individuals infected with human immunodeficiency virus type 1 (HIV-1). In the absence of KIR3DS1, the HLA-B Bw4-80Ile allele was not associated with any of the AIDS outcomes measured. By contrast, in the absence of HLA-B Bw4-80Ile alleles, KIR3DS1 was significantly associated with more rapid progression to AIDS. These observations are strongly suggestive of a model involving an epistatic interaction between the two loci. The strongest synergistic effect of these loci was on progression to depletion of CD4(+) T cells, which suggests that a protective response of NK cells involving KIR3DS1 and its HLA class I ligands begins soon after HIV-1 infection. C1 NCI, Basic Res Program, SAIC Frederick, Frederick, MD 21702 USA. NCI, Lab Genom Divers, Frederick, MD 21702 USA. Univ Calif Los Angeles, Sch Publ Hlth, Dept Epidemiol, Los Angeles, CA 90095 USA. NCI, Viral Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. San Francisco City Clin Cohort, San Francisco, CA 94102 USA. Univ Texas, Hlth Sci Ctr, Gulf States Hemophilia Ctr, Houston, TX 77030 USA. Johns Hopkins Med Inst, Sch Hyg & Publ Hlth, Baltimore, MD 21205 USA. Univ Cambridge, Dept Pathol, Div Immunol, Cambridge CB2 1QP, England. RP Carrington, M (reprint author), NCI, Basic Res Program, SAIC Frederick, Frederick, MD 21702 USA. NR 30 TC 734 Z9 771 U1 5 U2 25 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD AUG PY 2002 VL 31 IS 4 BP 429 EP 434 DI 10.1038/ng934 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 578XV UT WOS:000177147100022 PM 12134147 ER PT J AU Mykytyn, K Nishimura, DY Searby, CC Shastri, M Yen, HJ Beck, JS Braun, T Streb, LM Cornier, AS Cox, GF Fulton, AB Carmi, R Luleci, G Chandrasekharappa, SC Collins, FS Jacobson, SG Heckenlively, JR Weleber, RG Stone, EM Sheffield, VC AF Mykytyn, K Nishimura, DY Searby, CC Shastri, M Yen, HJ Beck, JS Braun, T Streb, LM Cornier, AS Cox, GF Fulton, AB Carmi, R Luleci, G Chandrasekharappa, SC Collins, FS Jacobson, SG Heckenlively, JR Weleber, RG Stone, EM Sheffield, VC TI Identification of the gene (BBS1) most commonly involved in Bardet-Biedl syndrome, a complex human obesity syndrome SO NATURE GENETICS LA English DT Article ID CHROMOSOME 16Q; SYNDROME LOCUS; DNA; MUTATIONS; INTERVAL; LINKAGE AB Bardet-Biedl syndrome (BBS, OMIM 209900) is a genetic disorder with the primary features of obesity, pigmentary retinopathy, polydactyly, renal malformations, mental retardation and hypogenitalism(1-4). Individuals with BBS are also at increased risk for diabetes mellitus, hypertension and congenital heart disease(4-6). What was once thought to be a homogeneous autosomal recessive disorder is now known to map to at least six loci: 11q13 (BBS1), 16q21 (BBS2), 3p13-p12 (BBS3), 15q22.3-q23 (BBS4), 2q31 (BBS5) and 20p12 (BBS6)(7-13). There has been considerable interest in identifying the genes that underlie BBS, because some components of the phenotype are common. Cases of BBS mapping ro BBS6 are caused by mutations in MKKS12,13; mutations in this gene also cause McKusick-Kaufman syndrome (hydrometrocolpos, post-axial polydactyly and congenital heart defects)(14,15). In addition, we recently used positional cloning to identify the genes underlying BBS2 (ref. 16) and BBS4 (ref. 17). The BBS6 protein has similarity to a Thermoplasma acidophilum chaperonin(15), whereas BBS2 and BBS4 have no significant similarity to chaperonins. It has recently been suggested that three mutated alleles (two at one locus, and a third at a second locus) may be required for manifestation of BBS (triallelic inheritance)(18). Here we report the identification of the gene BBS1 and show that a missense mutation of this gene is a frequent cause of BBS. In addition, we provide data showing that this common mutation is not involved in triallelic inheritance. C1 Univ Iowa, Dept Pediat, Div Med Genet, Iowa City, IA 52242 USA. Univ Iowa, Howard Hughes Med Inst, Iowa City, IA 52242 USA. Univ Iowa, Dept Elect Engn, Iowa City, IA 52242 USA. Univ Iowa, Dept Ophthalmol, Iowa City, IA 52242 USA. Ponce Sch Med, Dept Biochem, Ponce, PR USA. Childrens Hosp, Div Genet, Boston, MA 02115 USA. Childrens Hosp, Dept Ophthalmol, Boston, MA 02115 USA. Ben Gurion Univ Negev, Inst Genet, Soroka Med Ctr, IL-84105 Beer Sheva, Israel. Arkdeniz Univ, Dept Med Biol Genet, Antalya, Turkey. NHGRI, NIH, Bethesda, MD 20892 USA. Univ Penn, Scheie Eye Inst, Philadelphia, PA 19104 USA. Harbor UCLA Med Ctr, Jules Stein Eye Inst, Torrance, CA 90509 USA. Oregon Hlth Sci Univ, Casey Eye Inst, Portland, OR 97201 USA. RP Sheffield, VC (reprint author), Univ Iowa, Dept Pediat, Div Med Genet, Iowa City, IA 52242 USA. RI Mykytyn, Kirk/E-3723-2011; OI Mykytyn, Kirk/0000-0002-0158-4592; Searby, Charles/0000-0002-8108-8782 NR 22 TC 190 Z9 195 U1 1 U2 9 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD AUG PY 2002 VL 31 IS 4 BP 435 EP 438 DI 10.1038/ng935 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 578XV UT WOS:000177147100023 PM 12118255 ER PT J AU Kelsall, BL Biron, CA Sharma, O Kaye, PM AF Kelsall, BL Biron, CA Sharma, O Kaye, PM TI Dendritic cells at the host-pathogen interface SO NATURE IMMUNOLOGY LA English DT Editorial Material ID TOLL-LIKE RECEPTORS; MOUSE LYMPH-NODES; IN-VIVO; MYCOBACTERIUM-TUBERCULOSIS; T-CELLS; CD8-ALPHA(+); EXPRESSION; ACTIVATION; INFECTION; IMMUNITY C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Brown Univ, Dept Mol Microbiol & Immunol, Providence, RI 02912 USA. NIAID, Div AIDS, NIH, Bethesda, MD 20892 USA. London Sch Hyg & Trop Med, Dept Infect & Trop Dis, London WC1, England. RP Kelsall, BL (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 31 TC 107 Z9 110 U1 0 U2 3 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1529-2908 J9 NAT IMMUNOL JI Nat. Immunol. PD AUG PY 2002 VL 3 IS 8 BP 699 EP 702 DI 10.1038/ni0802-699 PG 4 WC Immunology SC Immunology GA 578LF UT WOS:000177118500003 PM 12145651 ER PT J AU Parravicini, V Gadina, M Kovarova, M Odom, S Gonzalez-Espinosa, C Furumoto, Y Saitoh, S Samelson, LE O'Shea, JJ Rivera, J AF Parravicini, V Gadina, M Kovarova, M Odom, S Gonzalez-Espinosa, C Furumoto, Y Saitoh, S Samelson, LE O'Shea, JJ Rivera, J TI Fyn kinase initiates complementary signals required for IgE-dependent mast cell degranulation SO NATURE IMMUNOLOGY LA English DT Article ID FC-EPSILON-RI; PROTEIN-TYROSINE KINASE; LYN-DEFICIENT MICE; HIGH-AFFINITY; T-CELL; AUTOIMMUNE-DISEASE; PLASMA-MEMBRANE; GAMMA-CHAIN; SH2 DOMAIN; ACTIVATION AB FcepsilonRI activation of mast cells is thought to involve Lyn and Syk kinases proximal to the receptor and the signaling complex organized by the linker for activation of T cells (LAT). We report here that FcepsilonRI also uses a Fyn kinase-dependent pathway that does not require Lyn kinase or the adapter LAT for its initiation, but is necessary for mast cell degranulation. Lyn-deficiency enhanced Fyn-dependent signals and degranulation, but inhibited the calcium response. Fyn-deficiency impaired degranulation, whereas Lyn-mediated signaling and calcium was normal. Thus, FcepsilonRI-dependent mast cell degranulation involves cross-talk between Fyn and Lyn kinases. C1 NCI, Cellular & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NIAMSD, Mol Inflammat Sect, NIH, Bethesda, MD 20892 USA. NIAMSD, Lymphocyte Cell Biol Sect, NIH, Bethesda, MD 20892 USA. Acad Sci Czech Republ, Inst Mol Genet, Prague, Czech Republic. RP Rivera, J (reprint author), IPN, CINVESTAV, Pharmacobiol Dept, Tenorios 235, Mexico City 14330, DF, Mexico. NR 49 TC 338 Z9 345 U1 2 U2 11 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1529-2908 J9 NAT IMMUNOL JI Nat. Immunol. PD AUG PY 2002 VL 3 IS 8 BP 741 EP 748 DI 10.1038/ni817 PG 8 WC Immunology SC Immunology GA 578LF UT WOS:000177118500013 PM 12089510 ER PT J AU Longo, DL AF Longo, DL TI DR's orders: Human antibody kills tumors by direct signaling SO NATURE MEDICINE LA English DT Editorial Material ID B-CELL LYMPHOMA; INHIBITION; ACTIVATION; GROWTH; BCL-2 AB Murine antibodies that block cell-surface antigens have been engineered ( humanized) to successfully elicit human immune-effector mechanisms. Now, a fully human antibody produced entirely in vitro kills tumor cells directly through signal transduction and shows promise against lymphoma in animal studies ( pages 801 - 807). C1 NIA, Baltimore, MD 21224 USA. RP Longo, DL (reprint author), NIA, Baltimore, MD 21224 USA. NR 12 TC 6 Z9 6 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1078-8956 J9 NAT MED JI Nat. Med. PD AUG PY 2002 VL 8 IS 8 BP 781 EP 783 DI 10.1038/nm0802-781 PG 4 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 579WJ UT WOS:000177200900017 PM 12152028 ER PT J AU Smolders, I Bortolotto, ZA Clarke, VRJ Warre, R Khan, GM O'Neill, MJ Ornstein, PL Bleakman, D Ogden, A Weiss, B Stables, JP Ho, KH Ebinger, G Collingridge, GL Lodge, D Michotte, Y AF Smolders, I Bortolotto, ZA Clarke, VRJ Warre, R Khan, GM O'Neill, MJ Ornstein, PL Bleakman, D Ogden, A Weiss, B Stables, JP Ho, KH Ebinger, G Collingridge, GL Lodge, D Michotte, Y TI Antagonists of GLU(K5)-containing kainate receptors prevent pilocarpine-induced limbic seizures SO NATURE NEUROSCIENCE LA English DT Article ID MOSSY FIBER SYNAPSES; SYNAPTIC TRANSMISSION; HIPPOCAMPAL-NEURONS; RAT HIPPOCAMPUS; KAINIC ACID; EPILEPTIFORM ACTIVITY; ANTIEPILEPTIC DRUGS; CEREBRAL-ISCHEMIA; PYRAMIDAL CELLS; GLUR5 SUBTYPE AB Developments in the molecular biology and pharmacology of GLU(K5), a subtype of the kainate class of ionotropic glutamate receptors, have enabled insights into the roles of this subunit in synaptic transmission and plasticity. However, little is known about the possible functions of GLU(K5)-containing kainate receptors in pathological conditions. We report here that, in hippocampal slices, selective antagonists of GLU(K5)-containing kainate receptors prevented development of epileptiform activity-evoked by the muscarinic agonist, pilocarpine-and inhibited the activity when it was preestablished. In conscious rats, these GLU(K5) antagonists prevented and interrupted limbic seizures induced by intra-hippocampal pilocarpine perfusion, and attenuated accompanying rises in extracellular L-glutamate and GABA. This anticonvulsant activity occurred without overt side effects. GLU(K5) antagonism also prevented epileptiform activity induced by electrical stimulation, both in vitro and in vivo. Therefore, we propose that subtype-selective GLU(K5) kainate receptor antagonists offer a potential new therapy for epilepsy. C1 Free Univ Brussels, Inst Pharmaceut, Dept Pharmaceut Chem & Drug Anal, B-1090 Brussels, Belgium. Univ Bristol, Sch Med, Dept Anat, MRC Ctr Synapt Plast, Bristol BS8 1TD, Avon, England. Eli Lilly & Co, Lilly Res Ctr, Windlesham GU20 6PH, Surrey, England. Eli Lilly & Co, Lilly Corp Ctr, Indianapolis, IN 46285 USA. NINDS, Epilepsy Branch, NIH, Bethesda, MD 20892 USA. Allelix Pharmaceut, Mississauga, ON L4V 1V7, Canada. Univ Hosp AZ VUB, Dept Neurol, B-1090 Brussels, Belgium. RP Lodge, D (reprint author), Free Univ Brussels, Inst Pharmaceut, Dept Pharmaceut Chem & Drug Anal, Laarbeeklaan 103, B-1090 Brussels, Belgium. RI Michotte, Yvette/G-9818-2013; Smolders, Ilse/J-9399-2013; Collingridge, Graham/C-4605-2015; Ornstein, Paul/E-4212-2016 OI Collingridge, Graham/0000-0002-9572-5359; Ornstein, Paul/0000-0002-4335-1877 NR 50 TC 101 Z9 105 U1 1 U2 12 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1097-6256 J9 NAT NEUROSCI JI Nat. Neurosci. PD AUG PY 2002 VL 5 IS 8 BP 796 EP 804 DI 10.1038/nn880 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 577XT UT WOS:000177088900024 PM 12080343 ER PT J AU Vousden, KH Lu, X AF Vousden, KH Lu, X TI Live or let die: The cell's response to p53 SO NATURE REVIEWS CANCER LA English DT Review ID WILD-TYPE P53; RIBONUCLEOTIDE REDUCTASE GENE; COLORECTAL-CANCER CELLS; MYC-INDUCED APOPTOSIS; NF-KAPPA-B; DNA-DAMAGE; P53-DEPENDENT APOPTOSIS; TRANSCRIPTIONAL TARGET; MUTANT P53; P53-INDUCED APOPTOSIS AB Compared with many normal tissues, cancer cells are highly sensitized to apoptotic signals, and survive only because they have acquired lesions - such as loss of p53 - that prevent or impede cell death. We are now beginning to understand the complex mechanisms that regulate whether or not a cell dies in response to p53 - insights that will ultimately contribute to the development of therapeutic strategies to repair the apoptotic p53 response in cancers. C1 NCI, Regulat Cell Growh Lab, Frederick, MD 21702 USA. Univ London Imperial Coll Sci Technol & Med, Ludwig Inst Canc Res, Fac Med, London W2 1PG, England. RP Vousden, KH (reprint author), Beatson Inst Canc Res, Garscube Estate,Switchback Rd, Glasgow G61 1BD, Lanark, Scotland. NR 145 TC 2086 Z9 2136 U1 17 U2 151 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1474-175X J9 NAT REV CANCER JI Nat. Rev. Cancer PD AUG PY 2002 VL 2 IS 8 BP 594 EP 604 DI 10.1038/nrc864 PG 11 WC Oncology SC Oncology GA 636GY UT WOS:000180447300013 PM 12154352 ER PT J AU Melino, G De Laurenzi, V Vousden, KH AF Melino, G De Laurenzi, V Vousden, KH TI p73: Friend or foe in tumorigenesis SO NATURE REVIEWS CANCER LA English DT Review ID WILD-TYPE P73; P53-RELATED PROTEIN P73; P53 HOMOLOG P73; KINASE C-ABL; CELL-LINES; SAM DOMAIN; CHROMOSOME 1P36.3; BREAST-CANCER; TRANSCRIPTIONAL ACTIVITIES; HEPATOCELLULAR-CARCINOMA AB As p53 and its homologue p73 have significant sequence and functional similarities, p73 might also be expected to act as a tumour suppressor However, p73 is activated after DNA damage in a way that is distinct from that of p53. The existence of DeltaNp73 - an isoform of p73 that is encoded by a distinct promoter and that lacks the transactivation domain - further complicates matters. It seems to function as an oncogene by inhibiting both p73- and p53-induced apoptosis. So how can these opposing functions be reconciled in human tumours?. C1 Univ Roma Tor Vergata, Biochem Lab, Dept Expt Med & Biochem Sci, I-00133 Rome, Italy. NCI, Regulat Cell Growth Lab, Frederick, MD 21702 USA. RP Melino, G (reprint author), Univ Roma Tor Vergata, Biochem Lab, Dept Expt Med & Biochem Sci, I-00133 Rome, Italy. FU Telethon [E.1224] NR 134 TC 416 Z9 427 U1 0 U2 10 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1474-175X J9 NAT REV CANCER JI Nat. Rev. Cancer PD AUG PY 2002 VL 2 IS 8 BP 605 EP 615 DI 10.1038/nrc861 PG 11 WC Oncology SC Oncology GA 636GY UT WOS:000180447300014 PM 12154353 ER PT J AU Nabel, EG AF Nabel, EG TI CDKS and CKIs: Molecular targets for tissue remodelling SO NATURE REVIEWS DRUG DISCOVERY LA English DT Review ID DEPENDENT-KINASE INHIBITOR; CELL-CYCLE ARREST; EXPERIMENTAL DIABETIC NEPHROPATHY; HEMATOPOIETIC STEM-CELLS; HUMAN CORONARY-ARTERIES; TUMOR-SUPPRESSOR GENE; GROWTH-FACTOR-BETA; MICE LACKING; TGF-BETA; NEOINTIMAL PROLIFERATION AB Effective tissue remodelling is essential to the survival of adult organs. Many of the signalling pathways that control these cellular decisions are regulated by nuclear interactions of cell-cycle proteins. Molecules that target cyclin-dependent kinases (CDKs) or CDK inhibitors (CKIs) represent a new class of therapeutic agents that influence tissue remodelling in several organ systems. An understanding of their cell-specific functions is leading to the development of exciting and bold approaches to the treatment cancer, cardiovascular disease and other diseases. C1 NHLBI, Cardiovasc Branch, NIH, Bethesda, MD 20892 USA. RP Nabel, EG (reprint author), NHLBI, Cardiovasc Branch, NIH, Bldg 10,8C103,10 Ctr Dr, Bethesda, MD 20892 USA. NR 139 TC 42 Z9 42 U1 0 U2 8 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1474-1776 J9 NAT REV DRUG DISCOV JI Nat. Rev. Drug Discov. PD AUG PY 2002 VL 1 IS 8 BP 587 EP 598 DI 10.1038/nrd869 PG 12 WC Biotechnology & Applied Microbiology; Pharmacology & Pharmacy SC Biotechnology & Applied Microbiology; Pharmacology & Pharmacy GA 607EJ UT WOS:000178778400012 PM 12402499 ER PT J AU Burke, B Stewart, CL AF Burke, B Stewart, CL TI Life at the edge: The nuclear envelope and human disease SO NATURE REVIEWS MOLECULAR CELL BIOLOGY LA English DT Review ID DREIFUSS MUSCULAR-DYSTROPHY; LAMINA-ASSOCIATED POLYPEPTIDE-2; FAMILIAL PARTIAL LIPODYSTROPHY; INTEGRAL MEMBRANE-PROTEINS; CAUSE AUTOSOMAL-DOMINANT; LEM-DOMAIN; CELL-CYCLE; A/C GENE; DILATED CARDIOMYOPATHY; MISSENSE MUTATIONS AB A group of human diseases, known as 'laminopathies', are associated with defects in proteins of the nuclear envelope. Most laminopathy mutations have been mapped to the A-type lamin gene, which is expressed in most adult cell types. So, why should different mutations in a near-ubiquitously expressed gene be associated with various discrete tissue-restricted diseases? Attempts to resolve this paradox are uncovering new molecular interactions - both inside the nucleus and at its periphery - which indicate that the nuclear envelope has functions that go beyond mere housekeeping. C1 Univ Florida, Dept Anat & Cell Biol, Gainesville, FL 32610 USA. NCI, Lab Canc & Dev Biol, FCRDC, Frederick, MD 21701 USA. RP Burke, B (reprint author), Univ Florida, Dept Anat & Cell Biol, 1600 SW Archer Rd, Gainesville, FL 32610 USA. NR 104 TC 290 Z9 304 U1 4 U2 20 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1471-0072 J9 NAT REV MOL CELL BIO JI Nat. Rev. Mol. Cell Biol. PD AUG PY 2002 VL 3 IS 8 BP 575 EP 585 DI 10.1038/nrm879 PG 11 WC Cell Biology SC Cell Biology GA 583UC UT WOS:000177427600017 PM 12154369 ER PT J AU Hirai, T Heymann, JAW Shi, D Sarker, R Maloney, PC Subramaniam, S AF Hirai, T Heymann, JAW Shi, D Sarker, R Maloney, PC Subramaniam, S TI Three-dimensional structure of a bacterial oxalate transporter SO NATURE STRUCTURAL BIOLOGY LA English DT Article ID 3-DIMENSIONAL STRUCTURE; MOLECULAR ARCHITECTURE; SCANNING MUTAGENESIS; ELECTRON-MICROSCOPY; ESCHERICHIA-COLI; SELECTIVITY; PROTEINS; CHANNEL AB The major facilitator superfamily (MFS) represents one of the largest classes of evolutionarily related membrane transporter proteins. Here we present the three-dimensional structure at 6.5 Angstrom resolution of a bacterial member of this superfamily, OAT. The structure, derived from an electron crystallographic analysis of two-dimensional crystals, reveals that the 12 helices in the OAT molecule are arranged around a central cavity, which is widest at the center of the membrane. The helices divide naturally into three groups: a peripheral set comprising helices 3, 6, 9 and 12; a second set comprising helices 2, 5, 8 and 11 that faces the central substrate transport pathway across most of the length of the membrane; and a third set comprising helices 1, 4, 7 and 10 that participate in the pathway either on the cytoplasmic side (4 and 10) or on the periplasmic side (1 and 7). Overall, the architecture of the protein is remarkably symmetric, providing a compelling molecular explanation for the ability of such transporters to carry out bi-directional substrate transport. C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Physiol, Baltimore, MD 21205 USA. RP Subramaniam, S (reprint author), NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RI Hirai, Teruhisa/G-2105-2015 OI Hirai, Teruhisa/0000-0002-2114-8149 NR 20 TC 163 Z9 166 U1 1 U2 3 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1072-8368 J9 NAT STRUCT BIOL JI Nat. Struct. Biol. PD AUG PY 2002 VL 9 IS 8 BP 597 EP 600 DI 10.1038/nsb821 PG 4 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 580BK UT WOS:000177214200012 PM 12118242 ER PT J AU Cui, YH Le, Y Zhang, X Gong, W Abe, K Sun, R Van Damme, J Proost, P Wang, JM AF Cui, YH Le, Y Zhang, X Gong, W Abe, K Sun, R Van Damme, J Proost, P Wang, JM TI Up-regulation of FPR2, a chemotactic receptor for amyloid beta 1-42 (A beta(42)), in murine microglial cells by TNF alpha SO NEUROBIOLOGY OF DISEASE LA English DT Article DE A beta(42); chemotaxis; FPR2; inflammation; microglia; TNF alpha; receptor ID TUMOR-NECROSIS-FACTOR; N-FORMYLPEPTIDE RECEPTOR; PROTEIN-COUPLED RECEPTOR; CHEMOKINE RECEPTORS; PEPTIDE RECEPTOR; MICE LACKING; IN-VITRO; EXPRESSION; NEUTROPHILS; ACTIVATION AB Human FPRL1 and its mouse homologue FPR2 are functional receptors for several exogenous and host-derived chemotactic peptides, including amyloid 1342 (A)342), a critical pathogenic factor in Alzheimer's disease. We investigated the effect of TNFalpha on the expression and function of FPR2 in mouse microglial cells, a crucial inflammatory cell type in the CNS. Primary murine microglia and a cell line N9 in resting state expressed low levels of FPR2 gene and lacked the response to chemotactic agonists for this receptor. Incubation with TNFa, however, increased microglial expression of FPR2 gene, in association with potent chemotactic responses to FPR2-specific agonists including Abeta(42). The effect of TNFalpha was dependent on the p55 TNFalpha receptor and activation of MAP kinase p38. TNFalpha concomitantly down-regulated microglial response to the chemokine SDF-1alpha. Thus, by selectively up-regulating FPR2 in microglia, TNFalpha has the capacity to amplify host response in inflammatory diseases in the CNS. (C) 2002 Elsevier Science (USA). C1 NCI, Canc Res Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NCI, Canc Res Ctr, Expt Immunol Lab, Frederick, MD 21702 USA. NCI, Intramural Res Support Program, SAIC Frederick, Frederick, MD 21702 USA. Univ Louvain, Rega Inst, Louvain, Belgium. RP Cui, YH (reprint author), NCI, Canc Res Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. RI Zhang, Xia/B-8152-2008 OI Zhang, Xia/0000-0002-9040-1486 FU NCI NIH HHS [N01-CO-56000] NR 45 TC 33 Z9 36 U1 0 U2 3 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0969-9961 J9 NEUROBIOL DIS JI Neurobiol. Dis. PD AUG PY 2002 VL 10 IS 3 BP 366 EP 377 DI 10.1006/nbdi.2002.0517 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 589GU UT WOS:000177751100016 PM 12270697 ER PT J AU Basile, AS AF Basile, AS TI Direct and indirect enhancement of GABAergic neurotransmission by ammonia: implications for the pathogenesis of hyperammonemic syndromes SO NEUROCHEMISTRY INTERNATIONAL LA English DT Article DE ammonia; muscimol; flunitrazepam; GABA(A) receptor; liver failure ID GAMMA-AMINOBUTYRIC-ACID; HEPATIC-ENCEPHALOPATHY; SYNAPTIC TRANSMISSION; RECEPTOR-BINDING; RAT-BRAIN; GABA; ASTROCYTES; MODULATION; GLUTAMATE; CHLORIDE AB Experimental evidence indicates that ammonia causes neuroexcitation and seizures. This contrasts with the lethargy, confusion and other manifestations of global CNS depression commonly considered to be major components of hyperammonemic encephalopathies. Substantial data now indicates that ammonia can modulate GABAergic neurotransmission through direct and indirect mechanisms. This modulation consists of an enhancement of GABAergic neurotransmission at concentrations commonly encountered in hyperammonemic states and precedes the suppression of inhibitory neuronal function observed at higher (>1 mM) ammonia concentrations. Not only is this increase in GABAergic neurotransmission consistent with the clinical picture of lethargy, ataxia and cognitive deficits associated with liver failure and congenital hyperammonemia, but it also provides a mechanism for testing new therapeutic modalities for the treatment of hyperammonemic encephalopathy. (C) 2002 Published by Elsevier Science Ltd. C1 NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Basile, AS (reprint author), NIDDK, Bioorgan Chem Lab, NIH, Bldg 8,Room 121,MSC 0826, Bethesda, MD 20892 USA. NR 36 TC 19 Z9 20 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0197-0186 J9 NEUROCHEM INT JI Neurochem. Int. PD AUG-SEP PY 2002 VL 41 IS 2-3 BP 115 EP 122 AR PII S0197-0186(02)00032-3 DI 10.1016/S0197-0186(02)00032-3 PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 576XP UT WOS:000177032700007 PM 12020612 ER PT J AU Langheim, FJP Callicott, JH Mattay, VS Duyn, JH Weinberger, DR AF Langheim, FJP Callicott, JH Mattay, VS Duyn, JH Weinberger, DR TI Cortical systems associated with covert music rehearsal SO NEUROIMAGE LA English DT Article DE fMRI; prefrontal; music; imagination; parietal ID POSITRON-EMISSION-TOMOGRAPHY; AUTOMATED IMAGE REGISTRATION; WORKING-MEMORY; FUNCTIONAL MRI; CORTEX ACTIVATION; BRAIN-REGIONS; PRIMARY MOTOR; PET; PERFORMANCE; FMRI AB Musical representation and overt music production are necessarily complex cognitive phenomena. While overt musical performance may be observed and studied, the act of performance itself necessarily skews results toward the importance of primary sensorimotor and auditory cortices. However, imagined musical performance (IMP) represents a complex behavioral task involving components suited to exploring the physiological underpinnings of musical cognition in music performance without the sensorimotor and auditory confounds of overt performance. We mapped the blood oxygenation level-dependent fMRI activation response associated with IMP in experienced musicians independent of the piece imagined. IMP consistently activated supplementary motor and premotor areas, right superior parietal lobule, right inferior frontal gyrus, bilateral mid-frontal gyri, and bilateral lateral cerebellum in contrast with rest, in a manner distinct from fingertapping versus rest and passive listening to the same piece versus rest. These data implicate an associative network independent of primary sensorimotor and auditory activity, likely representing the cortical elements most intimately linked to music production. C1 NIMH, Intramural Res Program, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. NINDS, Lab Funct & Mol Imaging, IRP, NIH, Bethesda, MD 20892 USA. RP Callicott, JH (reprint author), NIMH, Intramural Res Program, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. RI Duyn, Jozef/F-2483-2010; Callicott, Joseph/C-9102-2009 OI Callicott, Joseph/0000-0003-1298-3334 NR 47 TC 50 Z9 51 U1 0 U2 8 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1053-8119 J9 NEUROIMAGE JI Neuroimage PD AUG PY 2002 VL 16 IS 4 BP 901 EP 908 DI 10.1006/nimg.2002.1144 PG 8 WC Neurosciences; Neuroimaging; Radiology, Nuclear Medicine & Medical Imaging SC Neurosciences & Neurology; Radiology, Nuclear Medicine & Medical Imaging GA 584AK UT WOS:000177444900005 PM 12202078 ER PT J AU de Zwart, JA van Gelderen, P Kellman, P Duyn, JH AF de Zwart, JA van Gelderen, P Kellman, P Duyn, JH TI Reduction of gradient acoustic noise in MRI using SENSE-EPI SO NEUROIMAGE LA English DT Article ID AUDITORY-CORTEX; FMRI; SOUND; STIMULATION; MOTOR; COILS; NMR AB A new approach to reduce gradient acoustic noise levels in EPI experiments is presented. Using multichannel RF receive coils, combined. With SENSE data Acquisition and reconstruction, gradient slew-rates in single-shot EPI were reduced fourfold for rate-2 and ninefold for rate-3 SENSE. Multislice EPI experiments Were performed on three different scanner platforms. With 3.4 min in-plane resolution, measuring 6 slices per second (1 slices with 2000 ms TR), this resulted in average sound pressure level reductions of 11.3 dB(A) and 16.5 dB(A) for rate-2 and rate-3 SENSE, respectively. BOLD fMRI experiments, using visually paced finger-tapping paradigms, showed no detrimental effect of the acoustic noise reduction strategy con temporal noise levels and t scores. C1 NINDS, Lab Funct & Mol Imaging, NIH, Bethesda, MD 20892 USA. NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP de Zwart, JA (reprint author), NINDS, Lab Funct & Mol Imaging, NIH, Bldg 36,Rm 4D04, Bethesda, MD 20892 USA. RI Duyn, Jozef/F-2483-2010 NR 26 TC 40 Z9 40 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1053-8119 J9 NEUROIMAGE JI Neuroimage PD AUG PY 2002 VL 16 IS 4 BP 1151 EP 1155 DI 10.1006/nimg.2002.1119 PG 5 WC Neurosciences; Neuroimaging; Radiology, Nuclear Medicine & Medical Imaging SC Neurosciences & Neurology; Radiology, Nuclear Medicine & Medical Imaging GA 584AK UT WOS:000177444900028 PM 12202101 ER PT J AU Ikeda, SR Kammermeier, PJ AF Ikeda, SR Kammermeier, PJ TI M current mystery messenger revealed? SO NEURON LA English DT Editorial Material ID RAT SYMPATHETIC NEURONS; M-CHANNEL AB The identity of signaling elements that couple muscarinic acetylcholine receptor (mAChR) activation to M current (KCNQ K+ channels) modulation has remained unknown despite decades of study. Suh and Hille (in this issue of Neuron) demonstrate that activation of phospholipase C (PLC) initiates M current modulation and that recovery requires ATP and phosphoinositide 4-kinase (PI4-K). These data suggest that breakdown of phosphotidylinositol 4,5-bisphosphate (PIP2) is a crucial determinant of M channel modulation. C1 NIAAA, Lab Mol Physiol, NIH, Bethesda, MD 20892 USA. RP Ikeda, SR (reprint author), NIAAA, Lab Mol Physiol, NIH, Bethesda, MD 20892 USA. OI Ikeda, Stephen/0000-0002-4088-9508 NR 10 TC 9 Z9 10 U1 0 U2 0 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 0896-6273 J9 NEURON JI Neuron PD AUG 1 PY 2002 VL 35 IS 3 BP 411 EP 412 DI 10.1016/S0896-6273(02)00792-4 PG 2 WC Neurosciences SC Neurosciences & Neurology GA 582CB UT WOS:000177331200002 PM 12165463 ER PT J AU Luo, ZG Wang, Q Zhou, JZ Wang, JB Luo, ZJ Liu, MY He, X Wynshaw-Boris, A Xiong, WC Lu, B Mei, L AF Luo, ZG Wang, Q Zhou, JZ Wang, JB Luo, ZJ Liu, MY He, X Wynshaw-Boris, A Xiong, WC Lu, B Mei, L TI Regulation of AChR clustering by dishevelled interacting with MuSK and PAK1 SO NEURON LA English DT Article ID RECEPTOR TYROSINE KINASE; NICOTINIC ACETYLCHOLINE-RECEPTOR; NEUROMUSCULAR SYNAPSE FORMATION; POLARITY GENE; POSTSYNAPTIC APPARATUS; JUXTAMEMBRANE REGION; SIGNALING PATHWAYS; SKELETAL-MUSCLE; CELL POLARITY; CRD DOMAIN AB An important aspect of synapse development is the clustering of neurotransmitter receptors in the postsynaptic membrane. Although MuSK is required for acetylcholine receptor (AChR) clustering at the neuromuscular junction (NMJ), the underlying molecular mechanisms remain unclear. We report here that in muscle cells, MuSK interacts with Dishevelled (Dvl), a signaling molecule important for planar cell polarity. Disruption of the MuSK-Dvl interaction inhibits Agrin- and neuron-induced AChR clustering. Expression of dominant-negative Dvl1 in postsynaptic muscle cells reduces the amplitude of spontaneous synaptic currents at the NMJ. Moreover, Dvl1 interacts with downstream kinase PAK1. Agrin activates PAK, and this activation requires Dvl. Inhibition of PAK1 activity attenuates AChR clustering. These results demonstrate important roles of DO and PAK in Agrin/MuSK-induced AChR clustering and reveal a novel function of Dvl in synapse development. C1 Univ Alabama, Civitan Int Res Ctr, Dept Neurobiol, Birmingham, AL 35294 USA. Univ Alabama, Civitan Int Res Ctr, Dept Pathol, Birmingham, AL 35294 USA. Univ Alabama, Civitan Int Res Ctr, Dept Phys Med & Rehabil, Birmingham, AL 35294 USA. Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Neurosci, Shanghai 200031, Peoples R China. NICHD, Unit Synapse Dev & Plast, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. Univ Calif San Diego, Sch Med, Dept Pediat & Med, La Jolla, CA 92093 USA. Univ Calif San Diego, Sch Med, Ctr Canc, La Jolla, CA 92093 USA. Boston Univ, Sch Med, Evans Dept Med, Endocrinol Sect,Diabet & Metab Res Unit, Boston, MA 02118 USA. Boston Univ, Sch Med, Dept Biochem, Boston, MA 02118 USA. Texas A&M Univ Syst, Hlth Sci Ctr, Dept Med Biochem & Genet, Inst Biosci & Technol, Houston, TX 77030 USA. Harvard Univ, Sch Med, Childrens Hosp, Div Neurosci, Boston, MA 02115 USA. RP Mei, L (reprint author), Univ Alabama, Civitan Int Res Ctr, Dept Neurobiol, 1530 3rd Ave S, Birmingham, AL 35294 USA. RI Lu, Bai/A-4018-2012; Mei, Lin/G-8755-2012 NR 83 TC 157 Z9 163 U1 0 U2 4 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 0896-6273 J9 NEURON JI Neuron PD AUG 1 PY 2002 VL 35 IS 3 BP 489 EP 505 DI 10.1016/S0896-6273(02)00783-3 PG 17 WC Neurosciences SC Neurosciences & Neurology GA 582CB UT WOS:000177331200010 PM 12165471 ER PT J AU Tarasov, KV Tarasova, YS Crider, DG Anisimov, SV Wobus, AM Boheler, KR AF Tarasov, KV Tarasova, YS Crider, DG Anisimov, SV Wobus, AM Boheler, KR TI Galanin and galanin receptors in embryonic stem cells: accidental or essential? SO NEUROPEPTIDES LA English DT Article ID LEUKEMIA INHIBITORY FACTOR; SENSORY NEURONS; SELF-RENEWAL; TARGETED DISRUPTION; GENE-EXPRESSION; SERIAL ANALYSIS; LIGAND-BINDING; MECHANISMS; ACTIVATION; PATTERNS AB Galanin is a peptide consisting of 29 (mouse) or 30 (human) amino acids that was recently identified in undifferentiated mouse embryonic stem (ES) cells through transcriptome analyses. Galanin is known to have important modulatory roles in neuronal cells, but it is currently unclear what biological role, if any, galanin has in stem cells. Here we show that galanin transcripts represent a distinguishing molecular feature of embryonic stem cell lines and that all three galanin receptors subtypes are expressed in mouse ES cells (Gal-R2 > Gal-R3 much greater than Gal-R1). Based on cell culture data, galanin in a dose-dependent manner appears to regulate growth characteristics of ES cells, at least partially, through interactions with leukemia inhibitory factor (LIF), a cytokine implicated in the self-renewal process of ES cells. The regulation of ES cell growth can therefore be added to the list of biological processes regulated by this peptide. Published by Elsevier Science Ltd. C1 NIA, GRC, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. Inst Plant Genet, D-06466 Gatersleben, Germany. RP Boheler, KR (reprint author), NIA, GRC, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 35 TC 24 Z9 25 U1 0 U2 0 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0143-4179 J9 NEUROPEPTIDES JI Neuropeptides PD AUG PY 2002 VL 36 IS 4 BP 239 EP 245 DI 10.1016/S0143-4179(02)00050-1 PG 7 WC Endocrinology & Metabolism; Neurosciences SC Endocrinology & Metabolism; Neurosciences & Neurology GA 609BU UT WOS:000178883400001 PM 12372696 ER PT J AU Brenneman, DE Hauser, JM Spong, CY Phillips, TM AF Brenneman, DE Hauser, JM Spong, CY Phillips, TM TI Chemokine release is associated with the protective action of PACAP-38 against HIV envelope protein neurotoxicity SO NEUROPEPTIDES LA English DT Article ID VASOACTIVE-INTESTINAL-PEPTIDE; CYCLASE-ACTIVATING POLYPEPTIDE; IMMUNODEFICIENCY-VIRUS-INFECTION; FEMTOMOLAR CONCENTRATIONS; GP120 NEUROTOXICITY; CEREBRAL-CORTEX; RAT ASTROCYTES; CELL-CULTURES; COAT PROTEIN; HUMAN BRAIN AB The envelope protein (gp120) of the human immunodeficiency virus produces neuronal cell death in cultures that can be prevented by co-treatment with pituitary adenylate activating pepticle-38 (PACAP-38) or chemokines. To investigate the hypothesis that a functional relationship exists between these two protectants, the release of chemokines was measured in rat astrocyte cultures after PACAP-38 treatment. Chemokine analyses were performed by immunoaffinity capillary electrophoresis. Bell-shaped dose-responses for PACAP-mediated release of chemokines into the culture medium were observed with EC50's of 3 x 10(15) M (RANTES: regulated upon activation normal T cell expressed and secreted), 3 x 10(-11) M (MIP-1beta) and 10(-7) M (MIp-1alpha). In addition, PACAP-mediated depletion of chemokines from cultured astrocytes exhibited inverted bell-shaped curves, with similar EC50's to those observed for chemokine measurements of the medium. Comparative studies with structurally related peptides (vasoactive intestinal peptide [VIP] and secretin) revealed that PACAP was the most potent secretagogue for RANTES on astrocyte cultures. Gp120-mediated neuronal cell death was prevented by co-treatment with PACAP-38, although the efficacy of protection varied significantly among the gp120 isolates. A bi-model dose-response was observed with EC50's of 3 x 10(-15) and 3 x 10(-11) M. Co-treatment with neutralizing antiserum to RANTES attenuated PACAP-mediated protection from toxicity associated with gp120. In contrast to previous studies with VIP and gp120 toxicity, co-treatment with anti-MIP-1alpha did not affect PACAP-induced protection. These studies support the hypothesis that PACAP produces neuroprotection from gp120 toxicity, in part, through the release of RANTES and this mechanism is distinct from that observed with VIP. Published by Elsevier Science Ltd. C1 NICHHD, Dev Neurobiol Lab, Sect Dev & Mol Pharmacol, NIH, Bethesda, MD 20892 USA. NIH, Off Director, Off Res Serv, Div Bioengn & Phys Sci, Bethesda, MD 20892 USA. RP Brenneman, DE (reprint author), NICHHD, Dev Neurobiol Lab, Sect Dev & Mol Pharmacol, NIH, Bldg 49,Room 5A38, Bethesda, MD 20892 USA. NR 48 TC 35 Z9 36 U1 0 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0143-4179 J9 NEUROPEPTIDES JI Neuropeptides PD AUG PY 2002 VL 36 IS 4 BP 271 EP 280 DI 10.1016/S0143-4179(02)00045-8 PG 10 WC Endocrinology & Metabolism; Neurosciences SC Endocrinology & Metabolism; Neurosciences & Neurology GA 609BU UT WOS:000178883400006 PM 12372701 ER PT J AU Elman, I Goldstein, DS Green, AI Eisenhofer, G Folio, CJ Holmes, CS Pickar, D Breier, A AF Elman, I Goldstein, DS Green, AI Eisenhofer, G Folio, CJ Holmes, CS Pickar, D Breier, A TI Effects of risperidone on the peripheral noradrenegic system in patients with schizophrenia: A comparison with clozapine and placebo SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE schizophrenia; catecholamine; spillover; clearance; DHPG; tritium-labeled norepinephrine ID EARLY-ONSET SCHIZOPHRENIA; PLASMA NOREPINEPHRINE; NEGATIVE SYMPTOMS; TREATMENT RESPONSE; BIOGENIC-AMINES; HEART-FAILURE; CATECHOLAMINES; NORADRENALINE; NEUROLEPTICS; RELEASE AB Risperidone is an atypical antipsychotic drug that increases plasma norepinephrine (NE) levels, but the mechanism behind this effect is unclear. We measured arterial plasma levels of NE and other catechols during intravenous infusion of tritium-labeled NE (H-3-NE) in risperidone-treated patients and compared their data with those from patients treated with clozapine or placebo. NE levels in risperidone patients were significantly higher than in placebo patients, but lower than in clozapine patients. Neither drug, however, had significant effect on plasma levels of the main neuronal metabolite of NE, dihydroxyphenylglycol (DHPG), suggesting that adrenoceptors blockade alone would not explain the NE findings. The rate of release of endogenous NE into the bloodstream (spillover) was elevated in both risperidone and clozapine patients in a manner that paralleled their NE levels; the NE clearance in both groups did not differ from placebo. Following H-3-NE infusion in risperidone-treated individuals, production of H-3-DHPG was normal, as it was in the clozapine group, suggesting that risperidone does not impede neuronal uptake or intraneuronal metabolism of NE by monoamine oxidase. Our data suggest that both risperidone and clozapine elevate plasma NE levels via enhanced neurotransmitter spillover, with risperidone producing a smaller effect. C1 Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Psychiat, Boston, MA 02114 USA. Commonwealth Res Ctr, Boston, MA USA. NINDS, Clin Neurocardiol Sect, NIH, Bethesda, MD 20892 USA. Comprehens NeuroSci Inc, Washington, DC USA. Lilly Res Labs, Indianapolis, IN USA. Indiana Univ, Sch Med, Dept Psychiat, Indianapolis, IN 46202 USA. RP Elman, I (reprint author), Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Psychiat, 15 Parkman St WACC-812, Boston, MA 02114 USA. NR 63 TC 14 Z9 15 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD AUG PY 2002 VL 27 IS 2 BP 293 EP 300 AR PII S0893-133X(02)00314-7 DI 10.1016/S0893-133X(02)00314-7 PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 566AE UT WOS:000176402600017 PM 12093603 ER PT J AU Kenchaiah, S Evans, JC Levy, D Wilson, PWF Benjamin, EJ Larson, MG Kannel, WB Vasan, RS AF Kenchaiah, S Evans, JC Levy, D Wilson, PWF Benjamin, EJ Larson, MG Kannel, WB Vasan, RS TI Obesity and the risk of heart failure SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID LEFT-VENTRICULAR MASS; ARTERIAL-HYPERTENSION; GENERAL-POPULATION; WEIGHT-GAIN; FRAMINGHAM; DISEASE; MEN; PROGNOSIS; WOMEN AB Background Extreme obesity is recognized to be a risk factor for heart failure. It is unclear whether overweight and lesser degrees of obesity also pose a risk. Methods We investigated the relation between the body-mass index (the weight in kilograms divided by the square of the height in meters) and the incidence of heart failure among 5881 participants in the Framingham Heart Study (mean age, 55 years; 54 percent women). With the use of Cox proportional-hazards models, the body-mass index was evaluated both as a continuous variable and as a categorical variable (normal, 18.5 to 24.9; overweight, 25.0 to 29.9; and obese, 30.0 or more). Results During follow-up (mean, 14 years), heart failure developed in 496 subjects (258 women and 238 men). After adjustment for established risk factors, there was an increase in the risk of heart failure of 5 percent for men and 7 percent for women for each increment of 1 in body-mass index. As compared with subjects with a normal body-mass index, obese subjects had a doubling of the risk of heart failure. For women, the hazard ratio was 2.12 (95 percent confidence interval, 1.51 to 2.97); for men, the hazard ratio was 1.90 (95 percent confidence interval, 1.30 to 2.79). A graded increase in the risk of heart failure was observed across categories of body-mass index. The hazard ratios per increase in category were 1.46 in women (95 percent confidence interval, 1.23 to 1.72) and 1.37 in men (95 percent confidence interval, 1.13 to 1.67). Conclusions In our large, community-based sample, increased body-mass index was associated with an increased risk of heart failure. Given the high prevalence of obesity in the United States, strategies to promote optimal body weight may reduce the population burden of heart failure. C1 Framingham Heart Dis Epidemiol Study, Framingham, MA 01702 USA. Boston Univ, Sch Med, Sect Prevent Med, Boston, MA 02118 USA. Boston Univ, Sch Med, Cardiol Sect, Boston, MA 02118 USA. Beth Israel Deaconess Med Ctr, Div Cardiol, Boston, MA 02215 USA. Harvard Univ, Sch Med, Boston, MA USA. NHLBI, Bethesda, MD 20892 USA. RP Vasan, RS (reprint author), Framingham Heart Dis Epidemiol Study, 73 Mt Wayte Ave,Suite 2, Framingham, MA 01702 USA. RI Kenchaiah, Satish/A-1519-2016; OI Larson, Martin/0000-0002-9631-1254; Ramachandran, Vasan/0000-0001-7357-5970; Benjamin, Emelia/0000-0003-4076-2336 FU NHLBI NIH HHS [1K24 HL04334, N01-HC-25195] NR 37 TC 1189 Z9 1244 U1 6 U2 32 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD AUG 1 PY 2002 VL 347 IS 5 BP 305 EP 313 DI 10.1056/NEJMoa020245 PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 587WC UT WOS:000177665900002 PM 12151467 ER PT J AU McCracken, JT McGough, J Shah, B Cronin, P Hong, D Aman, MG Arnold, E Lindsay, R Nash, P Hollway, J McDougle, CJ Posey, D Swiezy, N Kohn, A Scahill, L Martin, A Koenig, K Volkmar, F Carroll, D Lancor, A Tierney, E Ghuman, J Gonzalez, NM Grados, M Vitiello, B Ritz, L Davies, M Robinson, J McMahon, D AF McCracken, JT McGough, J Shah, B Cronin, P Hong, D Aman, MG Arnold, E Lindsay, R Nash, P Hollway, J McDougle, CJ Posey, D Swiezy, N Kohn, A Scahill, L Martin, A Koenig, K Volkmar, F Carroll, D Lancor, A Tierney, E Ghuman, J Gonzalez, NM Grados, M Vitiello, B Ritz, L Davies, M Robinson, J McMahon, D CA Res Units Pediat Psychopharmacolog TI Risperidone in children with autism and serious behavioral problems SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID PERVASIVE DEVELOPMENTAL DISORDERS; ATYPICAL ANTIPSYCHOTICS; INFANTILE-AUTISM; HALOPERIDOL; ADOLESCENTS; SYMPTOMS; NETWORK; NORMS AB Background Atypical antipsychotic agents, which block postsynaptic dopamine and serotonin receptors, have advantages over traditional antipsychotic medications in the treatment of adults with schizophrenia and may be beneficial in children with autistic disorder who have serious behavioral disturbances. However, data on the safety and efficacy of atypical antipsychotic agents in children are limited. Methods We conducted a multisite, randomized, double-blind trial of risperidone as compared with placebo for the treatment of autistic disorder accompanied by severe tantrums, aggression, or self-injurious behavior in children 5 to 17 years old. The primary outcome measures were the score on the Irritability subscale of the Aberrant Behavior Checklist and the rating on the Clinical Global Impressions - Improvement (CGI-I) scale at eight weeks. Results A total of 101 children (82 boys and 19 girls; mean [+/-SD] age, 8.8 +/- 2.7 years) were randomly assigned to receive risperidone (49 children) or placebo (52). Treatment with risperidone for eight weeks (dose range, 0.5 to 3.5 mg per day) resulted in a 56.9 percent reduction in the Irritability score, as compared with a 14.1 percent decrease in the placebo group (P < 0.001). The rate of a positive response, defined as at least a 25 percent decrease in the Irritability score and a rating of much improved or very much improved on the CGI-I scale, was 69 percent in the risperidone group (34 of 49 children had a positive response) and 12 percent in the placebo group (6 of 52, P < 0.001). Risperidone therapy was associated with an average weight gain of 2.7 +/- 2.9 kg, as compared with 0.8 +/- 2.2 kg with placebo (P < 0.001). Increased appetite, fatigue, drowsiness, dizziness, and drooling were more common in the risperidone group than in the placebo group (P < 0.05 for each comparison). In two thirds of the children with a positive response to risperidone at eight weeks (23 of 34), the benefit was maintained at six months. Conclusions Risperidone was effective and well tolerated for the treatment of tantrums, aggression, or self-injurious behavior in children with autistic disorder. The short period of this trial limits inferences about adverse effects such as tardive dyskinesia. C1 Yale Univ, Ctr Child Study, New Haven, CT 06520 USA. Univ Calif Los Angeles, Los Angeles, CA 90024 USA. Ohio State Univ, Columbus, OH 43210 USA. Indiana Univ, Indianapolis, IN 46204 USA. Yale Univ, New Haven, CT USA. Kennedy Krieger Inst, Baltimore, MD USA. NIMH, Bethesda, MD 20892 USA. Columbia Univ, New York, NY USA. Nathan S Kline Inst Psychiat Res, Orangeburg, NY USA. RP Scahill, L (reprint author), Yale Univ, Ctr Child Study, POB 207900, New Haven, CT 06520 USA. OI Scahill, Lawrence/0000-0001-5073-1707 FU NCRR NIH HHS [M01 RR00052, M01 RR00034, M01 RR00750]; NIMH NIH HHS [K23 MH001883-02, N01MH70001, N01MH70009, N01MH70010, N01MH80011] NR 25 TC 703 Z9 721 U1 15 U2 53 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD AUG 1 PY 2002 VL 347 IS 5 BP 314 EP 321 DI 10.1056/NEJMoa013171 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 587WC UT WOS:000177665900003 PM 12151468 ER PT J AU Harger, JH Ernest, JM Thurnau, GR Moawad, A Thom, E Landon, MB Paul, R Miodovnik, M Dombrowski, M Sibai, B Van Dorsten, P McNellis, D AF Harger, JH Ernest, JM Thurnau, GR Moawad, A Thom, E Landon, MB Paul, R Miodovnik, M Dombrowski, M Sibai, B Van Dorsten, P McNellis, D CA Natl Inst Child Hlth Human Dev TI Frequency of congenital varicella syndrome in a prospective cohort of 347 pregnant women SO OBSTETRICS AND GYNECOLOGY LA English DT Article; Proceedings Paper CT 20th Annual Meeting of the Society-for-Maternal-Fetal-Medicine CY JAN 31-FEB 05, 2000 CL MIAMI BEACH, FLORIDA SP Soc Maternal Fetal Med ID MATERNAL VARICELLA; INFECTION AB OBJECTIVE: To estimate the rate of congenital varicella zoster virus syndrome in neonates born to women developing varicella zoster virus infections during pregnancy, METHODS: Pregnant women with clinical varicella zoster virus infection were enrolled at ten perinatal centers. Maternal and fetal immunoglobulin (Ig) G and IgM by fluorescent antibody confirmed 74.3% of cases. Specialists examined neonates at 0-6 months, 7-18 months, and 19-30 months after delivery to detect abnormalities of their eyes, hearing, and physical and developmental features. A hierarchical set of criteria was used to define congenital varicella syndrome. A jury of four investigators assigned the classification of all findings. RESULTS: In 362 women enrolled from 1993 to 1996, 15 had herpes zoster, and 347 had primary varicella zoster virus infection. Varicella zoster virus affected 140 women (38.7%) in the first trimester, 122 (133.7%) in the second trimester, and 100 (27.6%) in the third trimester. Five twin pairs were included. Only one case (0.4%) of definite congenital varicella syndrome was found, a 3360-g female infant having a left retinal macular lesion with typical skin scars after maternal varicella at 24 weeks. The maternal blood sample at birth was negative for IgG antibodies to toxoplasmosis and cytomegalovirus. Two cases involved fetal death at 20 weeks and fetal hydrops at 17 weeks after maternal varicella at 11 and 5 weeks, respectively. We found no cases of limb hypoplasia, microcephalus, or cataract. CONCLUSION: The frequency of congenital varicella syndrome is very low (0.4%) in a prospectively studied cohort. Eye examinations of exposed infants had a low yield. (C) 2002 by The American College of Obstetricians and Gynecologists. C1 Univ Pittsburgh, Sch Med, Dept Obstet Gynecol & Reprod Sci, Pittsburgh, PA 15213 USA. NICHHD, Bethesda, MD 20892 USA. RP Harger, JH (reprint author), Univ Pittsburgh, Sch Med, Dept Obstet Gynecol & Reprod Sci, 119 Douglas Dr, Pittsburgh, PA 15213 USA. FU NICHD NIH HHS [HD 21410, HD 27889, HD 27860, HD 27915, HD 27917, HD 27883, HD 19897, HD 27905, HD 27861, HD 21414] NR 16 TC 41 Z9 46 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD AUG PY 2002 VL 100 IS 2 BP 260 EP 265 AR PII S0029-7844(02)02059-8 DI 10.1016/S0029-7844(02)02059-8 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 578ZV UT WOS:000177151700010 PM 12151147 ER PT J AU Weaver, Z Montagna, C Xu, XL Howard, T Gadina, M Brodie, SG Deng, CX Ried, T AF Weaver, Z Montagna, C Xu, XL Howard, T Gadina, M Brodie, SG Deng, CX Ried, T TI Mammary tumors in mice conditionally mutant for Brca1 exhibit gross genomic instability and centrosome amplification yet display a recurring distribution of genomic imbalances that is similar to human breast cancer SO ONCOGENE LA English DT Article DE Brca1; mouse models; SKY; genomic imbalances; centrosome ID SUPPRESSOR GENE BRCA1; CHROMOSOMAL-ABERRATIONS; COPY NUMBER; CELL-CYCLE; MOUSE; TUMORIGENESIS; CARCINOMAS; CARCINOGENESIS; MORPHOGENESIS; PROGRESSION AB BRCA1 mutation carriers have an increased susceptibility to breast and ovarian cancer. Excision of exon 11 of Brca1 in the mouse, using a conditional knockout (Cre-loxP) approach, results in mammary tumor formation after long latency. To characterize the genomic instability observed in these tumors, to establish a comparative map of chromosomal imbalances and to contribute to the validation of this mouse model of breast cancer, we have characterized chromosomal imbalances and aberrations using comparative genomic hybridization (CGH), and spectral karyotyping (SKY). We found that all tumors exhibit chromosome instability as evidenced by structural chromosomal aberrations and aneuploidy, yet they display a pattern of chromosomal gain and loss that is similar to the pattern in human breast carcinomas. Of note, nine of 15 tumors exhibited a gain of distal chromosome 11, a region that is orthologous to human chromosome 17q11-qter, the mapping position of Erbb2. However, our analysis suggests that genes distal to Erbb2 are the main targets of amplification. Four of the tumors also exhibited a copy number loss of proximal chromosome 11 (11A-B), a region orthologous to human 17p. In eight of the tumors we observed whole or partial gain of chromosome 15 centering on 15D2-D3 (orthologous to human chromosome 8q24), the map location of the c-Myc gene, and six of the tumors exhibited copy number loss of whole or partial chromosome 14, including 14D3, the map location of Rb1. We conclude that despite the tremendous shuffling of chromosomes during the course of mammalian evolution, the pattern of genomic imbalances is conserved between BRCA1-associated mammary gland tumors in mice and humans. Western blot analysis showed that while p53 is absent or mutated in some tumors, at least two tumors revealed wild-type protein, suggesting that other genetic events may lead to tumorigenesis. Similar to BRCA1-deficient mouse embryonic fibroblasts, the tumor cells contained supernumerary functional centrosomes with intact centrioles whose presence results in multipolar mitoses and aneuploidy. C1 NIH, Genet Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NIDDK, Genet Dev & Dis Branch, NIH, Bethesda, MD USA. Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA. NIAMSD, NIH, Bethesda, MD USA. RP Ried, T (reprint author), NIH, Genet Branch, Ctr Canc Res, 50 S Dr,Room 1306, Bethesda, MD 20892 USA. RI deng, chuxia/N-6713-2016 NR 35 TC 97 Z9 103 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD AUG 1 PY 2002 VL 21 IS 33 BP 5097 EP 5107 DI 10.1038/sj.onc.1205636 PG 11 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 575YK UT WOS:000176975900008 PM 12140760 ER PT J AU Abrams, JS Schoenfeldt, M AF Abrams, JS Schoenfeldt, M TI Clinical Trials Referral Resource SO ONCOLOGY-NEW YORK LA English DT Editorial Material C1 NCI, Bethesda, MD 20892 USA. RP Abrams, JS (reprint author), NCI, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU P R R INC PI MELVILLE PA 48 SOUTH SERVICE RD, MELVILLE, NY 11747 USA SN 0890-9091 J9 ONCOLOGY-NY JI Oncology-NY PD AUG PY 2002 VL 16 IS 8 BP 1074 EP + PG 5 WC Oncology SC Oncology GA 585RR UT WOS:000177538800014 PM 12201646 ER PT J AU Rubinstein, WS O'Neill, SM Peters, JA Rittmeyer, LJ Stadler, MP AF Rubinstein, WS O'Neill, SM Peters, JA Rittmeyer, LJ Stadler, MP TI Mathematical modeling for breast cancer risk assessment - State of the art and role in medicine SO ONCOLOGY-NEW YORK LA English DT Editorial Material ID FAMILY HISTORY; OVARIAN-CANCER; BRCA2 MUTATIONS; INDIVIDUALIZED PROBABILITIES; PROPHYLACTIC MASTECTOMY; GENETIC-HETEROGENEITY; GERMLINE MUTATIONS; SEQUENCE-ANALYSIS; LIFE EXPECTANCY; WOMEN AB Women at increased risk of breast cancer have important opportunities for early detection and prevention. There are, however, serious drawbacks to the available interventions. The magnitude of breast cancer risk is a crucial factor in the optimization of medical benefit when considering the efficacy of risk-reduction methods, the adverse effects of intervention, and economic and quality-of-life outcomes. Breast cancer risk assessment has become increasingly quantitative and is amenable to computerization. The assembly of risk factor information into practical, quantitative models for clinical and scientific use is relatively advanced for breast cancer, and represents a paradigm for broader risk management in medicine. Using a case-based approach, we will summarize the major breast cancer risk assessment models, compare and contrast their utility, and illustrate the role of genetic testing in risk management. Important considerations relevant to clinical oncology practice include the role of risk assessment in cancer prevention, the logistics of implementing risk assessment, the ramifications of conveying risk information with limited genetic counseling, and the mechanisms for genetics referral. Medical professionals can embrace new preventive medicine techniques more effectively by utilizing quantitative methods to assess their patients' risks. C1 Evanston NW Healthcare, Ctr Med Genet, Evanston, IL 60201 USA. Northwestern Univ, Feinberg Sch Med, Evanston, IL USA. NCI, Clin Genet Branch, Div Canc Epidemiol & Genet, Rockville, MD USA. Univ Pittsburgh, Pittsburgh, PA USA. RP Rubinstein, WS (reprint author), Evanston NW Healthcare, Ctr Med Genet, 1000 Cent St,Suite 620, Evanston, IL 60201 USA. NR 56 TC 9 Z9 9 U1 0 U2 1 PU P R R INC PI MELVILLE PA 48 SOUTH SERVICE RD, MELVILLE, NY 11747 USA SN 0890-9091 J9 ONCOLOGY-NY JI Oncology-NY PD AUG PY 2002 VL 16 IS 8 BP 1082 EP 1094 PG 13 WC Oncology SC Oncology GA 585RR UT WOS:000177538800015 PM 12201647 ER PT J AU Goldberg, RM Saltz, L Grem, JL Lenz, HJ AF Goldberg, RM Saltz, L Grem, JL Lenz, HJ TI Oxaliplatin in colorectal cancer: New data in first-line and adjuvant therapy SO ONKOLOGIE LA German DT Article ID LEUCOVORIN; FLUOROURACIL; IRINOTECAN C1 Mayo Clin, Rochester, MN 55905 USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. NCI, Bethesda, MD 20892 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Wake Forest Univ, Winston Salem, NC 27109 USA. Univ Calif San Francisco, Ctr Canc, San Francisco, CA 94143 USA. Univ Iowa, Iowa City, IA 52242 USA. Univ Hosp Cleveland, Cleveland, OH 44106 USA. RP Goldberg, RM (reprint author), Mayo Clin, Rochester, MN 55905 USA. RI Goldberg , Richard/M-1311-2013 NR 20 TC 1 Z9 1 U1 0 U2 0 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 0378-584X J9 ONKOLOGIE JI Onkologie PD AUG PY 2002 VL 25 SU 3 BP 1 EP 11 PG 11 GA 599QQ UT WOS:000178347000002 ER PT J AU Gasch, AT Caruso, RC Kaler, SG Kaiser-Kupfer, M AF Gasch, AT Caruso, RC Kaler, SG Kaiser-Kupfer, M TI Menkes' syndrome - Ophthalmic findings SO OPHTHALMOLOGY LA English DT Article ID KINKY HAIR DISEASE; LYSYL OXIDASE AB Purpose: To report the prevalence and clinical significance of ocular findings in 20 patients with Menkes' syndrome recruited for a clinical trial at the National Institute of Child Health and Human Development (NICHHD). Design: Retrospective observational case series. Participants. Twenty patients with Menkes' syndrome enrolled in a clinical trial at the NICHHD, who underwent ophthalmic evaluation at the National Eye Institute from 1990 through 1997. Methods. Review of clinical ophthalmic examination records, photographs, and visual evoked potential recordings. Main Outcome Measures. Prevalence of ophthalmic signs of unusual frequency in patients with Menkes' syndrome. Results. There was a high prevalence of very poor visual acuity (8 patients), myopia at 21 months old or younger (5 of 9 patients this age who underwent cycloplegic refraction), strabismus (11 of 18 patients whose motility was evaluated), blue irides (15 of 16 patients whose iris color was documented), iris stromal hypoplasia and bilateral peripheral transillumination (7 and 3 patients, respectively), peripheral retinal hypopigmentation (7 of 18 patients whose retinal periphery was noted at the initial examination), and aberrant eyelashes (5 patients). Conclusions. Patients with Menkes' syndrome exhibit a high prevalence of several ocular findings, including some (very poor visual acuity, myopia, strabismus) that may warrant special care. Early ocular examination is thus indicated for patients with Menkes' syndrome, particularly those with mild variants of the disease, whose neurologic status is better and lifespan is longer. (C) 2002 by the American Academy of Ophthalmology. C1 Washington Hosp Ctr, Washington Natl Eye Ctr, Washington, DC 20010 USA. NINDS, Clin Neurosci Branch, NIH, Bethesda, MD 20892 USA. RP Kaiser-Kupfer, M (reprint author), NEI, Ophthalm Genet & Visual Funct Branch, NIH, 10 Ctr Dr,MSC 1860,Bldg 10,Room 10N226, Bethesda, MD 20892 USA. NR 33 TC 8 Z9 11 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0161-6420 J9 OPHTHALMOLOGY JI Ophthalmology PD AUG PY 2002 VL 109 IS 8 BP 1477 EP 1483 AR PII S0161-6420(02)01095-3 DI 10.1016/S0161-6420(02)01095-3 PG 7 WC Ophthalmology SC Ophthalmology GA 578ZR UT WOS:000177151400024 PM 12153799 ER PT J AU Takahashi, Y Ruhl, S Yoon, JW Sandberg, AL Cisar, JO AF Takahashi, Y Ruhl, S Yoon, JW Sandberg, AL Cisar, JO TI Adhesion of viridans group streptococci to sialic acid-, galactose- and N-acetylgalactosamine-containing receptors SO ORAL MICROBIOLOGY AND IMMUNOLOGY LA English DT Article DE adhesins; glycoconjugate receptors; viridans group streptococci ID GORDONII DL1 CHALLIS; CELL-WALL POLYSACCHARIDES; ACTINOMYCES-VISCOSUS T14V; TYPE-2 FIMBRIAL LECTIN; ORAL ACTINOMYCES; POLYMORPHONUCLEAR LEUKOCYTES; INTRAGENERIC COAGGREGATION; INSERTIONAL INACTIVATION; COLONIAL MORPHOLOGY; PHASE VARIATION AB The binding of 10 viridans group streptococci to sialic acid-, galactose (Gal)- and N-acetylgalactosamine (GalNAc)-containing receptors was defined by analysis of the interactions between these bacteria and structurally defined glycoconjugates, host cells and other streptococci. All interactions with sialic acid-containing receptors were Ca2+ -independent as they were not affected by ethyleneglycoltetraacetic acid (EGTA), whereas all interactions with Gal- and GalNAc-containing receptors were Ca2+-dependent. Recognition of sialic acid-, Gal- and GalNAc-containing receptors varied widely among the strains examined, in a manner consistent with the association of each of the three lectin-like activities with a different bacterial cell surface component. C1 NIDCR, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Cisar, JO (reprint author), NIDCR, Oral Infect & Immun Branch, NIH, Bldg 30,Room 532, Bethesda, MD 20892 USA. NR 35 TC 34 Z9 34 U1 0 U2 3 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0902-0055 J9 ORAL MICROBIOL IMMUN JI Oral Microbiol. Immunol. PD AUG PY 2002 VL 17 IS 4 BP 257 EP 262 DI 10.1034/j.1399-302X.2002.170409.x PG 6 WC Dentistry, Oral Surgery & Medicine; Immunology; Microbiology SC Dentistry, Oral Surgery & Medicine; Immunology; Microbiology GA 574TY UT WOS:000176906700009 PM 12121477 ER PT J AU Baum, BJ AF Baum, BJ TI Biomedical research, oral medicine, and the future SO ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTICS LA English DT Editorial Material C1 Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD USA. RP Baum, BJ (reprint author), Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 1079-2104 J9 ORAL SURG ORAL MED O JI Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. PD AUG PY 2002 VL 94 IS 2 BP 141 EP 142 DI 10.1067/moe.2002.127564 PG 2 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 592VG UT WOS:000177957100001 PM 12221377 ER PT J AU Sankar, V Brennan, MT Radfar, L Leakan, RA Pillemer, SR AF Sankar, V Brennan, MT Radfar, L Leakan, RA Pillemer, SR TI Elevated blood pressure is not related to saliva flow in patients with Sjogren's syndrome SO ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTICS LA English DT Article ID MUSCARINIC ACETYLCHOLINE-RECEPTORS; AUTOANTIBODIES; HYPERTENSION; ANTIBODIES; CLASSIFICATION; NEUROPATHY; CRITERIA; GLANDS; ALPHA; RATES AB Objective. The purpose of this study was to determine whether systolic and diastolic blood pressures are associated with salivary flow, dry mouth, or dry eye symptoms in patients with primary Sjogren's syndrome as compared with xerostomic control subjects. Study design. One hundred forty consecutive patients seen at the Sjogren's Syndrome Clinic were categorized retrospectively with various classification schemes: (1) subjective dry mouth; (2) subjective dry eye; (3) European criteria; and (4) international criteria. Data collection included age, gender, systolic blood pressure, diastolic blood pressure, salivary flow rate, focus score, Schirmer's test, and laboratory findings, including antinuclear antibodies, anti-SSA, anti-SSB, IgG, IgA, IgM, erythrocyte sedimentation rate, and rheumatoid factor. Results. No meaningful associations of salivary flow rates with systolic or diastolic blood pressures were found in patients with Sjogren's syndrome or in xerostomic control subjects. An inverse correlation was seen between salivary flow and elevated diastolic blood pressure in xerostomic control subjects only. Conclusion. Elevated blood pressure was not related to saliva flow in patients with Sjogren's syndrome. C1 NIDCR, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA. Carolinas Med Ctr, Dept Oral Med, Charlotte, NC 28203 USA. NIH, Clin Ctr Nursing, Bethesda, MD 20892 USA. RP Sankar, V (reprint author), NIDCR, Gene Therapy & Therapeut Branch, NIH, Bldg 10,Rm 1N113,10 Ctr Dr,MSC 1190, Bethesda, MD 20892 USA. NR 33 TC 4 Z9 4 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 1079-2104 J9 ORAL SURG ORAL MED O JI Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. PD AUG PY 2002 VL 94 IS 2 BP 179 EP 183 DI 10.1067/moe.2002.123861 PG 5 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 592VG UT WOS:000177957100009 PM 12221385 ER PT J AU Han, F Kipnes, JR Li, Y Tuan, RS Hall, DJ AF Han, F Kipnes, JR Li, Y Tuan, RS Hall, DJ TI The murine COMP (cartilage oligomeric matrix protein) promoter contains a potent transcriptional repressor region SO OSTEOARTHRITIS AND CARTILAGE LA English DT Article DE COMP; cartilage; arthritis; genomic clone; promoter; transcription ID MULTIPLE EPIPHYSEAL DYSPLASIA; 5-STRANDED COILED-COIL; PSEUDOACHONDROPLASIA; MUTATIONS; GENE; IDENTIFICATION; EXPRESSION AB Objective: A subgroup of patients with pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED) have been found to harbor mutations within the cartilage oligomeric matrix protein (COMP) gene. These two diseases are autosomal dominant disorders that are characterized by an early onset of osteoarthritis (OA). The COMP gene is expressed primarily in chondrocytes in articular cartilage as well as in tendon and ligament. Therefore, control over tissue specific COMP expression may be an important aspect in cartilage biology. To begin an analysis of the regulation of COMP expression, we have cloned, sequenced and characterized the entire genomic clone for mouse COMP that includes the COMP promoter. Methods and Results: The COMP coding region spans 19 exons over approximately 8.4 kb of DNA. The arrangement and size of the exons have a remarkable similarity to those of the human COMP genomic sequence, indicating a significant degree of genomic conservation. Analysis of a 453 basepair region of the putative COMP promoter reveals two strong transcriptional repressor elements located between position -356 and -304 and between -251 and -180, relative to the start site for transcription. These repressor elements down-regulate transcription from the promoter in a broad spectrum of cell lines. Removal of the repressor DNA sequence from the COMP promoter leads to significant enhancement in transcriptional activity, indicating that this region acts in a dominant manner to transcriptional activators located more proximal to the start site of transcription. This region also represses transcription when linked to a heterologous promoter. Conclusions: This repressor region probably down-regulates transcription from the COMP promoter in vivo. It may help to repress transcription of COMP in non-cartilaginous tissues and/or may aid in the expression of COMP to the appropriate level in tissues such as cartilage, tendon and ligament. (C) 2002 Elsevier Science Ltd on behalf of OsteoArthritis Research Society International. C1 NIAMSD, Cartilage Biol & Orthopaed Branch, NIH, Bethesda, MD 20892 USA. Harvard Univ, Sch Dent Med, Dept Growth & Dev, Boston, MA 02115 USA. RP Hall, DJ (reprint author), NIAMSD, Cartilage Biol Branch, NIH, MSC 5755,Bldg 13,Rm 3W17, Bethesda, MD 20892 USA. FU NIAMS NIH HHS [AR39740, AR45823] NR 25 TC 6 Z9 6 U1 1 U2 2 PU W B SAUNDERS CO LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1063-4584 J9 OSTEOARTHR CARTILAGE JI Osteoarthritis Cartilage PD AUG PY 2002 VL 10 IS 8 BP 638 EP 645 DI 10.1053/joca.2002.0532 PG 8 WC Orthopedics; Rheumatology SC Orthopedics; Rheumatology GA 594GD UT WOS:000178040100007 PM 12479386 ER PT J AU Stoll, BJ Hansen, N Fanaroff, AA Wright, LL Carlo, WA Ehrenkranz, RA Lemons, JA Donovan, EF Stark, AR Tyson, JE Oh, W Bauer, CR Korones, SB Shankaran, S Laptook, AR Stevenson, DK Papile, LA Poole, WK AF Stoll, BJ Hansen, N Fanaroff, AA Wright, LL Carlo, WA Ehrenkranz, RA Lemons, JA Donovan, EF Stark, AR Tyson, JE Oh, W Bauer, CR Korones, SB Shankaran, S Laptook, AR Stevenson, DK Papile, LA Poole, WK TI Late-onset sepsis in very low birth weight neonates: The experience of the NICHD Neonatal Research Network SO PEDIATRICS LA English DT Article DE sepsis; infant; newborn infant; very low birth weight infant; premature ID INTENSIVE-CARE UNIT; BLOOD-STREAM INFECTIONS; INFANTS AB Objective. Late-onset sepsis (occurring after 3 days of age) is an important problem in very low birth weight (VLBW) infants. To determine the current incidence of late-onset sepsis, risk factors for disease, and the impact of late-onset sepsis on subsequent hospital course, we evaluated a cohort of 6956 VLBW (401-1500 g) neonates admitted to the clinical centers of the National Institute of Child Health and Human Development Neonatal Research Network over a 2-year period (1998-2000). Methods. The National Institute of Child Health and Human Development Neonatal Research Network maintains a prospective registry of all VLBW neonates admitted to participating centers within 14 days of birth. Expanded infection surveillance was added in 1998. Results. Of 6215 infants who survived beyond 3 days, 1313 (21%) had 1 or more episodes of blood culture-proven late-onset sepsis. The vast majority of infections (70%) were caused by Gram-positive organisms, with coagulase-negative staphylococci accounting for 48% of infections. Rate of infection was inversely related to birth weight and gestational age. Complications of prematurity associated with an increased rate of late-onset sepsis included patent ductus arteriosus, prolonged ventilation, prolonged intravascular access, bronchopulmonary dysplasia, and necrotizing enterocolitis. Infants who developed late-onset sepsis had a significantly prolonged hospital stay (mean length of stay: 79 vs 60 days). They were significantly more likely to die than those who were uninfected (18% vs 7%), especially if they were infected with Gram-negative organisms (36%) or fungi (32%). Conclusions. Late-onset sepsis remains an important risk factor for death among VLBW preterm infants and for prolonged hospital stay among VLBW survivors. Strategies to reduce late-onset sepsis and its medical, social, and economic toll need to be addressed urgently. C1 Emory Univ, Sch Med, Dept Pediat, Atlanta, GA 30322 USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. Case Western Reserve Univ, Dept Pediat, Cleveland, OH 44106 USA. NICHHD, Bethesda, MD 20892 USA. Univ Alabama, Dept Pediat, Birmingham, AL USA. Yale Univ, Sch Med, Dept Pediat, New Haven, CT 06510 USA. Indiana Univ Sch Med, Dept Pediat, Indianapolis, IN USA. Univ Cincinnati, Coll Med, Dept Pediat, Cincinnati, OH 45267 USA. Harvard Univ, Childrens Hosp, Joint Program Neonatol, Boston, MA 02115 USA. Univ Texas, Houston Med Sch, Hlth Sci Ctr, Ctr Clin Res & Evidence Based Med, Houston, TX USA. Brown Univ, Dept Pediat, Providence, RI 02912 USA. Univ Miami, Dept Pediat, Miami, FL 33152 USA. Univ Tennessee, Newborn Ctr, Memphis, TN USA. Wayne State Univ, Div Neonatal & Perinatal Med, Detroit, MI USA. Univ Texas, SW Med Ctr, Dept Pediat, Dallas, TX USA. Stanford Univ, Med Ctr, Div Neonatol, Palo Alto, CA 94304 USA. Univ New Mexico, Sch Med, Dept Pediat, Albuquerque, NM 87131 USA. RP Stoll, BJ (reprint author), Emory Univ, Sch Med, Dept Pediat, 2040 Ridgewood Dr NE, Atlanta, GA 30322 USA. FU NICHD NIH HHS [U01 HD36790, U10 HD21364, U10 HD21373, U10 HD21385, U10 HD21397, U10 HD21415, U10 HD27851, U10 HD27853, U10 HD27856, U10 HD27871, U10 HD27880, U10 HD27881, U10 HD27904, U10 HD34167, U10 HD34216, U10 HD40689] NR 20 TC 967 Z9 1027 U1 6 U2 38 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD AUG PY 2002 VL 110 IS 2 BP 285 EP 291 DI 10.1542/peds.110.2.285 PG 7 WC Pediatrics SC Pediatrics GA 581DA UT WOS:000177274800021 PM 12165580 ER PT J AU Salbe, AD Weyer, C Lindsay, RS Ravussin, E Tataranni, PA AF Salbe, AD Weyer, C Lindsay, RS Ravussin, E Tataranni, PA TI Assessing risk factors for obesity between childhood and adolescence: I. Birth weight, childhood adiposity, parental obesity, insulin, and leptin SO PEDIATRICS LA English DT Article DE childhood obesity; growth and development; parent-child relationship; overweight tracking; Pima Indians ID BODY-MASS INDEX; PHYSICAL-ACTIVITY; PIMA-INDIANS; 5-YEAR-OLD CHILDREN; DIABETES-MELLITUS; YOUNG ADULTHOOD; FOLLOW-UP; OVERWEIGHT; GAIN; PREVALENCE AB Objective. To assess the effects of body weight, body composition, parental obesity, and metabolic variables on the development of obesity in a large cohort of 5-year-old Native American children with a high propensity for obesity. Methods. During the summer months of 1992 to 1995 and again 5 years later, 138 (65 boys and 73 girls) 5-year-old Pima Indian children were studied. Height; weight; body composition; parental obesity; and fasting plasma insulin, glucose, and leptin concentrations were determined at baseline and follow-up. Linear regression models were used to assess the effect of the baseline variables on the development of obesity. Results. At both 5 and 10 years of age, Pima Indian children were heavier and fatter than an age- and gender-matched reference population. All anthropometric and metabolic variables tracked strongly from 5 to 10 years of age (r greater than or equal to 0.70). The most significant determinant of percentage of body fat at 10 years of age was percentage of body fat at 5 years of age (R-2 = 0.53). The combined effect of high maternal body mass index, elevated fasting plasma leptin concentrations, and low fasting plasma insulin concentrations at baseline explained an additional 4% of the total variance in adiposity at follow-up. Conclusions. Although parental obesity and metabolic variables such as insulinemia and leptinemia at baseline account for a small percentage of the variance in adiposity at follow-up, early childhood obesity is the dominant predictor of obesity 5 years later. These results suggest that strategies to prevent childhood obesity must be initiated at a very early age. C1 NIDDKD, NIH, CDNS, Phoenix, AZ 85016 USA. NIDDKD, Clin Diabet & Nutr Sect, NIH, Bethesda, MD 20892 USA. NIDDKD, Diabet Arthritis Epidemiol Sect, NIH, Bethesda, MD 20892 USA. Louisiana State Univ, Pennington Biomed Res Ctr, Baton Rouge, LA 70808 USA. RP Salbe, AD (reprint author), NIDDKD, NIH, CDNS, 4212 N 16th St, Phoenix, AZ 85016 USA. NR 53 TC 74 Z9 75 U1 0 U2 7 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD AUG PY 2002 VL 110 IS 2 BP 299 EP 306 DI 10.1542/peds.110.2.299 PG 8 WC Pediatrics SC Pediatrics GA 581DA UT WOS:000177274800023 PM 12165582 ER PT J AU Salbe, AD Weyer, C Harper, I Lindsay, RS Ravussin, E Tataranni, PA AF Salbe, AD Weyer, C Harper, I Lindsay, RS Ravussin, E Tataranni, PA TI Assessing risk factors for obesity between childhood and adolescence: II. Energy metabolism and physical activity SO PEDIATRICS LA English DT Article DE childhood obesity; growth and development; energy metabolism; physical activity; Pima Indians ID DOUBLY-LABELED WATER; RESPIRATORY EXCHANGE RATIO; BODY-WEIGHT GAIN; 5-YEAR-OLD CHILDREN; CAUCASIAN CHILDREN; AFRICAN-AMERICAN; FUEL UTILIZATION; WHITE-CHILDREN; YOUNG-CHILDREN; UNITED-STATES AB Objective. To assess the effect of energy expenditure, including resting metabolic rate (RMR), total energy expenditure (TEE), and activity energy expenditure (AEE), as well as substrate oxidation (respiratory quotient [RQ]), on the development of obesity in a large cohort of Native American children with a high propensity for obesity. Methods. During the summer months of 1992 to 1995 and again 5 years later, 138 (65 boys and 73 girls) 5-year-old Pima Indian children were studied. At baseline and follow-up, height and weight were measured; body composition was assessed with the use of O-18 dilution; RMR and RQ were assessed with the use of indirect calorimetry; TEE was measured with the use of the doubly-labeled water method; and AEE was calculated (TEE - [RMR + 0.1 x TEE]). In addition, an activity questionnaire was used to assess participation in sporting activities as well as television viewing during the previous year. Linear regression models were used to assess the effects of the baseline variables on the development of obesity. Results. Pima Indian children were markedly overweight at both 5 and 10 years of age. Cross-sectionally, percentage of body fat and body weight at 5 and 10 years of age were negatively correlated with sports participation and positively correlated with television viewing. Most important, there was a marked change in the correlation between body size and activity between 5 and 10 years of age: at age 5 years, weight was positively correlated with AEE and PAL, but at age 10 years, the correlation with AEE was lost and that with PAL was negative. However, prospectively, none of the variables measured at baseline was a predictor of percentage of body fat at age 10 years after adjustment for percentage of body fat at age 5 years. Conclusions. At age 5 years, obesity is associated with decreased participation in sports and increased television viewing but not with a decreased PAL. At age 10 years, obesity is associated with decreased participation in sports, increased television viewing, and a decreased PAL, suggesting that a decrease in PAL in free-living conditions seems to follow, not precede, the development of obesity. C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Bethesda, MD 20892 USA. NIDDKD, Diabet & Arthritis Epidemiol Sect, NIH, Bethesda, MD 20892 USA. Louisiana State Univ, Pennington Biomed Res Ctr, Baton Rouge, LA 70808 USA. RP Salbe, AD (reprint author), NIDDKD, NIH, CDNS, 4212 N 16th St, Phoenix, AZ 85016 USA. NR 62 TC 78 Z9 80 U1 1 U2 14 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD AUG PY 2002 VL 110 IS 2 BP 307 EP 314 DI 10.1542/peds.110.2.307 PG 8 WC Pediatrics SC Pediatrics GA 581DA UT WOS:000177274800024 PM 12165583 ER PT J AU Snider, LA Seligman, LD Ketchen, BR Levitt, SJ Bates, LR Garvey, MA Swedo, SE AF Snider, LA Seligman, LD Ketchen, BR Levitt, SJ Bates, LR Garvey, MA Swedo, SE TI Tics and problem behaviors in schoolchildren: Prevalence, characterization, and associations SO PEDIATRICS LA English DT Article DE observational study; longitudinal study; tic disorder; childhood onset ID TOURETTES-SYNDROME; DISORDERS AB Objective. Tic disorders are the most common movement disorder diagnosed in children and have symptoms that fluctuate in frequency and intensity over time. We conducted an 8-month longitudinal observational study to determine the variations in frequency of motor tics and associated problem behaviors. Methods. A total of 553 children, kindergarten through sixth grade, were observed monthly from November 1999 to June 2000 by 3 raters. Motor tics were recorded by location and rated for severity as none (0), mild (1), moderate (2), or severe (3). Problem behaviors were rated as absent (0), subclinical (1), or clinical (2) in each of 6 categories: disruptive, hyperactive, impulsive, aggressive, anxious, and distracted. Results. The monthly point prevalence of motor tics ranged from 3.2% to 9.6%, with an overall frequency of 24.4%. The monthly point prevalence of problem behaviors ranged from 2.6% to 11.0%, with an overall frequency of 25.7%. The incidence of motor tics and problem behaviors was significantly higher during the winter months of November through February, compared with the spring months of March through June (motor tics: z = 4.97; problem behaviors: z = 3.79). Motor tics were observed in 2 distinct patterns (isolated and persistent), which varied by the number of months present, gender ratio of affected children, severity of tic symptoms, and association with problem behaviors. Conclusions. Motor tics and problem behaviors are frequent occurrences among schoolchildren and seem to occur more frequently during the winter months. For most children, the tics were mild, observed on only 1 occasion, and were not accompanied by problem behaviors. C1 NIMH, Pediat & Dev Neuropsychiat Branch, NIH, Bethesda, MD 20892 USA. RP Snider, LA (reprint author), NIMH, Pediat & Dev Neuropsychiat Branch, NIH, 10 Ctr Dr,Room 4N208,MSC 1255, Bethesda, MD 20892 USA. NR 20 TC 61 Z9 66 U1 0 U2 1 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD AUG PY 2002 VL 110 IS 2 BP 331 EP 336 DI 10.1542/peds.110.2.331 PG 6 WC Pediatrics SC Pediatrics GA 581DA UT WOS:000177274800027 PM 12165586 ER PT J AU Shankaran, S Laptook, A Wright, LL Ehrenkranz, RA Donovan, EF Fanaroff, AA Stark, AR Tyson, JE Poole, K Carlo, WA Lemons, JA Oh, W Stoll, BJ Papile, LA Bauer, CR Stevenson, DK Korones, SB McDonald, S AF Shankaran, S Laptook, A Wright, LL Ehrenkranz, RA Donovan, EF Fanaroff, AA Stark, AR Tyson, JE Poole, K Carlo, WA Lemons, JA Oh, W Stoll, BJ Papile, LA Bauer, CR Stevenson, DK Korones, SB McDonald, S TI Whole-body hypothermia for neonatal encephalopathy: Animal observations as a basis for a randomized, controlled pilot study in term infants SO PEDIATRICS LA English DT Article DE encephalopathy; hypothermia; asphyxia; term infants ID BRAIN INJURY; POSTISCHEMIC HYPOTHERMIA; NEWBORN-INFANTS; SELECTIVE HEAD; BLOOD-FLOW; NEUROPROTECTION; ISCHEMIA; TEMPERATURE; PROTECTION; ASPHYXIA AB Objective. Modest reduction in brain temperature is a promising therapy to reduce brain damage after neonatal encephalopathy as a result of acute perinatal asphyxia. The efficacy of modest hypothermia may in part be dependent on the stability of the desired brain temperature. The objective of this study was 1) to evaluate in newborn animals a commercially available cooling system (Blanketrol II Hyperthermia-Hypothermia system) to control brain temperature during whole-body hypothermia and 2) to use the results of the animal experiments to perform a pilot study evaluating the feasibility of whole-body hypothermia as a neuroprotective therapy for newborns with encephalopathy at birth. Methods. In the animal investigation, 3 miniature swine were instrumented and ventilated, and temperature probes were placed in the esophagus and the brain (1 cm and 2 cm beneath the parietal cortical surface and the dura). Body cooling was achieved using the automatic control mode (servo) of the cooling system. In the human investigation, 19 term infants with moderate or severe encephalopathy were randomized to either normothermia (n = 10) or hypothermia (n = 9) within 6 hours of birth. Whole-body hypothermia was achieved using the hyperthermia-hypothermia cooling system with servo control of esophageal temperature to 34.5degrees C for 72 hours followed by slow rewarming. Results. In the animal investigation, body cooling with the animal lying on a single blanket resulted in rapid cooling of the body within 90 minutes. Repetitive cyclical swings in esophageal temperature of 1.7 +/- 0.2degrees C (mean +/- standard deviation) around the set point of 33.5degrees C were reduced to 0.7 +/- 0.2degrees C when a second, larger blanket was attached and suspended. Esophageal temperature was a good marker of deep brain temperature (esophageal to 2-cm brain difference: 0.1 +/- 0.3degrees C). In the human investigation, the infants were randomized at 4.1 +/- 1.3 hours (mean +/- standard deviation) after birth. Age at randomization was similar in the 2 groups. Cooling was initiated at an average age of 5.3 hours. Target temperature of 34.5degrees C was achieved within 30 minutes and remained constant throughout the intervention period. Heart rate decreased to 108 +/- 14 beats per minute (bpm) at 60 minutes and remained between 115 and 130 bpm for the duration of cooling compared with 130 to 145 bpm in the normothermia group. Blood pressure was similar in the 2 groups. No adverse events occurred during 72 hours of cooling. The mortality rate and frequency of persistent pulmonary hypertension, renal failure, hepatic dysfunction, and need for pressor support were similar in both groups. Conclusions. Animal studies showed that a simple modification of a commercially available cooling system (2 blankets attached, subject lying on 1 and the second hanging freely) results in stable core body and brain temperature when used in the automatic control mode. The pilot study in term infants with encephalopathy using this cooling system demonstrates feasibility of initiating whole-body hypothermia at <6 hours of age to a constant esophageal temperature using servo control and provides no evidence that hypothermia involved greater hazard than benefit. C1 Wayne State Univ, Detroit, MI USA. Univ Texas, SW Med Ctr, Dallas, TX USA. NICHHD, Bethesda, MD 20892 USA. Yale Univ, New Haven, CT USA. Univ Cincinnati, Cincinnati, OH USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. Brigham & Womens Hosp, Boston, MA 02115 USA. Univ Texas, Sch Med, Houston, TX USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. Univ Alabama, Birmingham, AL USA. Indiana Univ, Indianapolis, IN 46204 USA. Brown Univ, Women & Infants Hosp, Providence, RI USA. Emory Univ, Atlanta, GA 30322 USA. Univ New Mexico, Albuquerque, NM 87131 USA. Univ Miami, Miami, FL 33152 USA. Stanford Univ, Stanford, CA 94305 USA. Univ Tennessee, Memphis, TN USA. RP Shankaran, S (reprint author), Childrens Hosp Michigan, 3901 Beaubien, Detroit, MI 48201 USA. FU NICHD NIH HHS [U10 HD34167, U01 HD36790, U10 HD21364, U10 HD21373, U10 HD21385, U10 HD21397, U10 HD21415, U10 HD27851, U10 HD27853, U10 HD27856, U10 HD27871, U10 HD27880, U10 HD27881, U10 HD27904, U10 HD34216, U10 HD40689] NR 25 TC 131 Z9 135 U1 0 U2 1 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD AUG PY 2002 VL 110 IS 2 BP 377 EP 385 DI 10.1542/peds.110.2.377 PG 9 WC Pediatrics SC Pediatrics GA 581DA UT WOS:000177274800035 PM 12165594 ER PT J AU Jia, L Wong, H Cerna, C Weitman, SD AF Jia, L Wong, H Cerna, C Weitman, SD TI Effect of nanonization on absorption of 301029: Ex vivo and in vivo pharmacokinetic correlations determined by liquid chromatography/mass spectrometry SO PHARMACEUTICAL RESEARCH LA English DT Article DE nanonization; permeability; bioavailability; thiadiazole; ex vivo-in vivo correlation ID NITRIC-OXIDE DONOR; PHARMACODYNAMICS; MITOXANTRONE; TRANSPORT; CELLS; MICE AB Purpose. To compare Caco-2 monolayer permeability and in vivo bioavailability of microparticle with nanoparticle 301029, a thiadiazole derivative, and to determine whether nanonization could improve oral bioavailability of the poorly soluble compound. Methods. The mean particle size of 301029 was reduced from 7 mum to 280 nm by pearl milling. In the ex vivo assay, both microparticle and nanoparticle 301029 at the same concentration were separately added to apical side and were collected from basolateral side of Caco-2 monolayer. In the bioavailability study, the two particle sizes of 301029 were orally administered to rats, respectively, and blood samples were collected. Nanoparticle 301029 in culture medium and rat serum was detected by a liquid chomatography-mass spectrometer (LC/MS) coupled with atmospheric pressure chemical ionization (APCI). Results. Permeability rate and permeated amounts of nanoparticle 301029 across the Caco-2 monolayer were about four times higher than those of microparticle 301029. In a pharmacokinetic study, nanoparticle 301029 showed T-max about 1 h, whereas the microparticle 301029 showed T-max at 4 h. The C-max and AUC of nanoparticle 301029 were 3- to 4-fold greater than those of microparticle 301029, resulting in a significant increase in oral bioavailability of 301029 as compared with microparticle 301029. The ex vivo permeability and in vivo pharmacokinetic data indicate that nanoparticle formulation improves both absorption rate and absorption extent of the poorly soluble drug. Conclusions. Nanoparticle formulation enhances both Caco-2 monolayer permeability and rat oral bioavailability of the poorly soluble 301029. The result also demonstrates a close correlation between ex vivo Caco-2 permeability model and in vivo gastrointestinal absorption. C1 Canc Therapy & Res Ctr S Texas, Inst Drug Dev, San Antonio, TX 78245 USA. Ilex Oncol Inc, San Antonio, TX 78245 USA. RP Jia, L (reprint author), NIH, EPN, Rm 8042,6130 Execut Blvd, Rockville, MD 20852 USA. NR 14 TC 50 Z9 51 U1 1 U2 5 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0724-8741 J9 PHARMACEUT RES JI Pharm. Res. PD AUG PY 2002 VL 19 IS 8 BP 1091 EP 1096 AR UNSP 0724-8741/02/0800-1091/0 DI 10.1023/A:1019829622088 PG 6 WC Chemistry, Multidisciplinary; Pharmacology & Pharmacy SC Chemistry; Pharmacology & Pharmacy GA 586MN UT WOS:000177587900002 PM 12240933 ER PT J AU Shalev, U Kafkafi, N AF Shalev, U Kafkafi, N TI Repeated maternal separation does not alter sucrose-reinforced and open-field behaviors SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE emotionality; male; maternal separation; open-field; operant response; rat; reward; sucrose preference ID PROGRESSIVE RATIO SCHEDULE; STRESSFUL LIFE EVENT; ADULT-RATS; EXPLORATORY-BEHAVIOR; BENZODIAZEPINE RECEPTOR; RATTUS-NORVEGICUS; D-AMPHETAMINE; ANIMAL-MODEL; RESPONSES; DOPAMINE AB Repeated separation of rat pups from their mothers has been reported to increase behavioral fearfulness and hypothalamic-pituitary-adrenal (HPA) response to stress. Recently, it was suggested that it might also alter behavioral responses to natural and drug rewards. Here, we studied whether maternal separation (MS) would alter behavioral responses to a sucrose reward. We also tested whether MS would alter behavioral responses in an open-field test using a novel method of analysis [Software for the Exploration of Exploration (SEE)]. Long-Evans rat pups were exposed to either 180 min of MS, 15 min of separation [early handling (EH)] or left undisturbed [nonhandled (NH)] from postnatal day (PND) 3 to 14. The adult male offspring were tested for sucrose solution preference using a two-bottle free-choice test, operant response for sucrose under fixed ratio and progressive ratio (PR) schedules of reinforcement and response to a novel environment (open-field test). MS had no effect on sucrose preference or operant responding for sucrose reward. In the open-field test, NH rats showed a brief decrease in locomotor response, but MS rats did not differ from the NH and EH groups in the other behavioral measures. Thus, under the conditions of the present study, MS did not appear to alter reward-related processes and also had a minimal effect on open-field behavior. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NIDA, Behav Neurosci Branch, IRP, NIH, Baltimore, MD 21224 USA. RP Shalev, U (reprint author), NIDA, Behav Neurosci Branch, IRP, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 46 TC 57 Z9 59 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD AUG PY 2002 VL 73 IS 1 BP 115 EP 122 AR PII S0091-3057(02)00756-6 DI 10.1016/S0091-3057(02)00756-6 PG 8 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 575KR UT WOS:000176945900012 PM 12076730 ER PT J AU Makino, S Hashimoto, K Gold, PW AF Makino, S Hashimoto, K Gold, PW TI Multiple feedback mechanisms activating corticotropin-releasing hormone system in the brain during stress SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Review DE corticotropin-releasing hormone; paraventricular nucleus; amygdala; bed nucleus of the stria terminalis; vasopressin; corticosteroid receptor; neuropeptide Y; antidepressant ID RECEPTOR MESSENGER-RNA; HYPOTHALAMIC PARAVENTRICULAR NUCLEUS; PITUITARY-ADRENOCORTICAL AXIS; FACTOR-LIKE IMMUNOREACTIVITY; CENTRAL AMYGDALOID NUCLEUS; ANXIETY-LIKE BEHAVIOR; REPEATED IMMOBILIZATION STRESS; II CORTICOSTEROID RECEPTORS; RAT-BRAIN; STRIA TERMINALIS AB Stress-associated disorders such as melancholic depression are characterized by persistent hypothalamic-pituitary-adrenocortical (HPA) axis activation and intensive anxiety. Corticotropin-releasing hormone (CRH) appears to play an essential role in pathophysiology of such disorders. In an attempt to elucidate possible mechanisms underlying persistent activation of CRH in the central nervous system (CNS), we examined responses of hypothalamic and extrahypothalamic CRH systems to the stressors (immobilization stress or psychological stress) and interactions between these CRH systems and glucocorticoids in rats. We propose multiple feedback loops activating central CRH system: (1) attenuation of glucocorticoid-induced negative feedback on the activity of the hypothalamic and brainstem nuclei during chronic stress, (2) autoregulation of CRH biosynthesis in the hypothalamic paraventricular nucleus (PVN) through up-regulation of Type-1 CRH receptor (CRHR-1), and (3) glucocorticoid-mediated positive effects on the amygdaloid CRH system. Stress initially activates the hypothalamic CRH system, resulting in the hypersecretion of glucocorticoids from the adrenal gland. In addition, the psychological component of the stressor stimulates the amygdaloid CRH system. In the chronic phase of stress, down-regulation of GR in the PVN and other brain structures such as the locus coeruleus (LC) fails to restrain hyperfunction of the HPA axis, and persistent activation of the HPA axis further up-regulates the amygdaloid CRH system. Thus, the hypothalamic and the amygdaloid CRH systems cooperatively constitute stress-responsive, anxiety-producing neurocircuitry during chronic stress, which is responsible for the clinical manifestations of stress-associated disorders. Effects of tricyclic antidepressants (TCAs), which appear to mitigate the above mentioned multiple feedback loop forming the vicious circle to activate central CRH systems, will also be discussed. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Kochi Med Sch, Dept Internal Med 2, Kochi 783, Japan. NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. RP Makino, S (reprint author), Kochi Med Sch, Dept Internal Med 2, Okoh Cho, Kochi 783, Japan. NR 160 TC 160 Z9 168 U1 2 U2 19 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD AUG PY 2002 VL 73 IS 1 BP 147 EP 158 AR PII S0091-3057(02)00791-8 DI 10.1016/S0091-3057(02)00791-8 PG 12 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 575KR UT WOS:000176945900016 PM 12076734 ER PT J AU Hall, FS Ghaed, S Pert, A Xing, G AF Hall, FS Ghaed, S Pert, A Xing, G TI The effects of isolation rearing on glutamate receptor NMDAR1A mRNA expression determined by in situ hybridization in Fawn hooded and Wistar rats SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE isolation rearing; Fawn hooded rat; glutamate receptors; NMDA; in situ hybridization ID VENTRAL TEGMENTAL AREA; ACOUSTIC STARTLE RESPONSE; MEDIAL PREFRONTAL CORTEX; FORCED SWIM TEST; NUCLEUS-ACCUMBENS; PREPULSE INHIBITION; SOCIAL-ISOLATION; LOCOMOTOR-ACTIVITY; ADULT RATS; ENVIRONMENTAL ENRICHMENT AB Rats reared in social isolation exhibit a syndrome of behavioral and biochemical effects indicative of enhanced mesolimbic dopamine (DA) function. The precise nature of the neurodevelopmental changes that produce this state are unknown but result in enhanced DA neurotransmission in the nucleus accumbens (NAC). It was hypothesized that this may be the indirect result of chronic changes in glutamate NMDA receptor function. The same prediction has been made for Fawn hooded (FH) rats that exhibit some of the characteristic effects of isolation-reared rats when compared to Wistar rats. Therefore, mRNA levels of the NMDAR1A receptor subunit were determined by in situ hybridization and were quantified in the striatum, hippocampus and prefrontal cortex of FH and Wistar rats. Isolation rearing alone was not found to have an effect on the expression of NMDAR1A, while FH rats had reduced levels across most brain regions examined. In some areas of the striatum and prefrontal cortex, this effect was greater in FH isolates than in FH socials, while in the hippocampus, the opposite was observed. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NIDA, Mol Neurobiol Branch, IRP, Baltimore, MD 21224 USA. Univ Maryland, Dept Psychol, College Pk, MD 20742 USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. RP Hall, FS (reprint author), NIDA, Mol Neurobiol Branch, IRP, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Hall, Frank/C-3036-2013 OI Hall, Frank/0000-0002-0822-4063 NR 58 TC 23 Z9 23 U1 0 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD AUG PY 2002 VL 73 IS 1 BP 185 EP 191 AR PII S0091-3057(02)00796-7 DI 10.1016/S0091-3057(02)00796-7 PG 7 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 575KR UT WOS:000176945900020 PM 12076738 ER PT J AU Gubbins, PO Penzak, SR Polston, S McConnell, SA Anaissie, E AF Gubbins, PO Penzak, SR Polston, S McConnell, SA Anaissie, E TI Characterizing and predicting amphotericin B-associated nephrotoxicity in bone marrow or peripheral blood stem cell transplant recipients SO PHARMACOTHERAPY LA English DT Article; Proceedings Paper CT 38th Interscience Conference on Antimicrobial Agents and Chemotherapy CY SEP 24-28, 1998 CL SAN DIEGO, CALIFORNIA ID RISK-FACTORS AB Study Objective. To characterize amphotericin B-associated nephrotoxicity and to determine the variables associated with it that can be used to identify, a priori, at-risk patients. Design. Retrospective analysis. Setting. University hospital. Patients. A homogeneous population of 69 recipients of a bone marrow (BMT) or peripheral blood stem cell transplant (PBSCT) with multiple myeloma and who received at least two doses of amphotericin B deoxycholate from January 1, 1992-January 1, 1995. Intervention. Data on demographics, prior and concomitant nephrotoxic drug therapy, daily, laboratory values, and amphotericin B dosing were collected serially from medical and pharmacy records. Measurements and Main Results. Forward stepwise logistic regression analysis was performed on the data from the first day of therapy to characterize and determine variables related to amphotericin B-associated nephrotoxicity. Nephrotoxicity occurred in 30 patients (43%) and developed rapidly Patients who developed nephrotoxicity were similar to those who did not in many aspects associated with their treatment. However, baseline estimated creatinine clearance, cyclosporine therapy, nephrotoxic drug therapy within 30 days of starting amphotericin B, and the number of concomitant nephrotoxic drugs were significant predictors of amphotericin B-associated nephrotoxicity. Conclusion. Recipients of a BMT or PBSCT who have multiple myeloma and are receiving cyclosporine or multiple nephrotoxic drugs at the start of amphotericin B therapy should be considered at high risk for developing amphotericin B-associated nephrotoxicity. Also, because amphotericin B-associated nephrotoxicity develops rapidly, clinicians should be aware of the rapid changes in serum creatinine and electrolyte levels that can occur. C1 Univ Arkansas Med Sci, Coll Pharm, Little Rock, AR 72205 USA. Univ Arkansas Med Sci, Myeloma Inst Res & Therapy, Little Rock, AR 72205 USA. NIH, Clin Pharmacokinet Lab, Dept Pharm, Ctr Clin, Bethesda, MD USA. Cent Arkansas Hosp, Dept Pharm, Searcy, AR USA. RP Gubbins, PO (reprint author), Univ Arkansas Med Sci, Coll Pharm, 4301 W Markham,Slot 522, Little Rock, AR 72205 USA. NR 16 TC 22 Z9 23 U1 0 U2 0 PU PHARMACOTHERAPY PUBLICATIONS INC PI BOSTON PA NEW ENGLAND MEDICAL CENTER, 806, 750 WASHINGTON ST, BOSTON, MA 02111 USA SN 0277-0008 J9 PHARMACOTHERAPY JI Pharmacotherapy PD AUG PY 2002 VL 22 IS 8 BP 961 EP 971 DI 10.1592/phco.22.12.961.33607 PG 11 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 580AQ UT WOS:000177212200004 PM 12173799 ER PT J AU Rachel, RA Mason, CA Beermann, F AF Rachel, RA Mason, CA Beermann, F TI Influence of tyrosinase levels on pigment accumulation in the retinal pigment epithelium and on the uncrossed retinal projection SO PIGMENT CELL RESEARCH LA English DT Article DE dark-eyed albino (Tyr(c44H) ); melanin; retinal ganglion cells; tyrosinase minigene ID GANGLION-CELL PROJECTIONS; TRANSGENIC MICE; OPTIC CHIASM; ALBINO MICE; OCULAR ALBINISM; GENETIC-CONTROL; MOLECULAR-BASIS; MOUSE; EXPRESSION; PROTEINS AB To study the relationship among tyrosinase activity, melanin production, and the routing of retinal ganglion cell (RGC) axons at the optic chiasm, we analysed mice with varying doses of the tyrosinase gene. These include the dark-eyed albino (Tyr(c44H)), a radiation-induced hypomorphic allele of tyrosinase; and transgenic mice carrying 1 or 2 alleles of a tyrosinase minigene on both wild-type (Tyr(+)) and albino (Tyr(c)) backgrounds. Melanization of the retinal pigment epithelium (RPE) occurred gradually even at <2% wild-type tyrosinase activity and was sensitive to tyrosinase activity up to <35% of wild-type levels, beyond which melanin synthesis appeared to be saturated. Overexpression of tyrosinase led to tyrosinase activity above wild type level, but did not increase melanin production. Although a loss of melanin because of a mutation in tyrosinase is associated with a decrease in the number of uncrossed fibers, elevating tyrosinase levels does not appear to cause an increase in the size of the uncrossed retinal projection. Our results suggest that replacing less than 35% of wild-type tyrosinase activity is sufficient to restore normal pigmentation of the RPE, and potentially, to allay visual defects. C1 Columbia Univ Coll Phys & Surg, Ctr Neurobiol & Behav, New York, NY 10032 USA. Columbia Univ Coll Phys & Surg, Dept Pathol, New York, NY 10032 USA. Columbia Univ Coll Phys & Surg, Dept Anat & Cell Biol, New York, NY 10032 USA. Swiss Inst Expt Canc Res, CH-1066 Epalinges, Switzerland. RP Rachel, RA (reprint author), NCI, Mouse Canc Genet Program, POB B,Bldg 539,Room 234, Frederick, MD 21702 USA. FU NEI NIH HHS [EY12736, R01 EY015290] NR 41 TC 20 Z9 20 U1 1 U2 3 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0893-5785 J9 PIGM CELL RES JI Pigm. Cell. Res. PD AUG PY 2002 VL 15 IS 4 BP 273 EP 281 DI 10.1034/j.1600-0749.2002.02019.x PG 9 WC Cell Biology; Dermatology SC Cell Biology; Dermatology GA 570MN UT WOS:000176662200005 PM 12100493 ER PT J AU Kalache, KD Bierlich, A Hammer, H Bollmann, R AF Kalache, KD Bierlich, A Hammer, H Bollmann, R TI Is unexplained third trimester intrauterine death of fetuses with gastroschisis caused by umbilical cord compression due to acute extra-abdominal bowel dilatation? SO PRENATAL DIAGNOSIS LA English DT Article DE gastroschisis; prenatal diagnosis; Doppler ultrasonography ID VELOCITY WAVE-FORMS; FETAL AB We report on a case of gastroschisis in which sudden dilatation of extra-abdominal bowel at 34+1 weeks was followed by an unusual umbilical flow velocity waveform (diastolic notching). The condition was associated with normal umbilical Doppler indices, brain sparing effect and a non-reactive cardiotocograph (CTG). Findings at postnatal surgery strongly suggested severe cord compression by the herniated dilated bowel. The significance of notching in the umbilical artery waveform is discussed, as is the potential importance of this sign in the prepartum management of fetuses with gastroschisis. Copyright (C) 2002 John Wiley Sons, Ltd. C1 Humboldt Univ Hosp, Fetal Med Unit, Dept Obstet & Gynecol, Fac Med, Berlin, Germany. Humboldt Univ Hosp, Fac Med, Clin Neonatol, Berlin, Germany. RP Kalache, KD (reprint author), Wayne State Univ, Hutzel Hosp, Perinatol Res Branch, NICHD,NIH, 4707 St Antoine Blvd, Detroit, MI 48201 USA. NR 9 TC 20 Z9 20 U1 0 U2 0 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0197-3851 J9 PRENATAL DIAG JI Prenat. Diagn. PD AUG PY 2002 VL 22 IS 8 BP 715 EP 717 DI 10.1002/pd.386 PG 3 WC Genetics & Heredity; Obstetrics & Gynecology SC Genetics & Heredity; Obstetrics & Gynecology GA 583ZY UT WOS:000177443300014 PM 12210582 ER PT J AU Siahpush, M Singh, GK AF Siahpush, M Singh, GK TI Sociodemographic variations in breast cancer screening behavior among Australian women: Results from the 1995 National Health Survey SO PREVENTIVE MEDICINE LA English DT Article DE breast cancer screening; sociodemographic; Australia ID VIETNAMESE-AMERICAN WOMEN; MAMMOGRAPHY USE; PRIMARY-CARE; SELF-REPORT; BARRIERS; AUDIT; AGE AB Background. Knowledge of sociodemographic variations in breast cancer screening can help identify population groups that are at risk of underutilization of breast cancer screening procedures and practices. We examined sociodemographic variations in breast cancer screening behavior among Australian women. Methods. We used a subsample of women aged 18 years and older (n = 10,179) from the 1995 National Health Survey. We examined the association of sociodemogarphic variables with mammography, clinical breast examination, and breast self-examination. Results. Being in the oldest age group, never being or previously being married, living in rural regions (except in the case of breast self-examination), residing in more disadvantaged areas (except in the case of breast self-examination), and having lower levels of education were all associated with a smaller likelihood of screening. Ethnicity was also significantly associated with screening. Conclusion. Strategies to promote breast cancer screening practices should pay particular attention to the underserved groups and should be part of a more comprehensive policy that ensures the accessibility to regular health care of these population groups. (C) 2002 American Health Foundation and Elsevier Science (USA). C1 Canc Council Victoria, Vichlth Ctr Tobacco Control, Carlton, Vic 3053, Australia. NCI, NIH, Bethesda, MD 20892 USA. RP Siahpush, M (reprint author), Canc Council Victoria, Vichlth Ctr Tobacco Control, 100 Drummond St, Carlton, Vic 3053, Australia. NR 46 TC 47 Z9 49 U1 2 U2 3 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0091-7435 J9 PREV MED JI Prev. Med. PD AUG PY 2002 VL 35 IS 2 BP 174 EP 180 DI 10.1006/pmed.2002.1063 PG 7 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 583GA UT WOS:000177397400012 PM 12200103 ER PT J AU Lin, Y Salem, N AF Lin, Y Salem, N TI A technique for the in vivo study of multiple stable isotope-labeled essential fatty acids SO PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS LA English DT Article ID ARACHIDONIC-ACID; DOCOSAHEXAENOIC ACIDS; LINOLENIC ACIDS; IN-VIVO; METABOLISM; HUMANS; INFANTS; CONVERSION; FORMULAS; RATS AB A novel tracer technique is presented for the simultaneous and independent measurement of multiple stable isotopically labeled essential fatty acids. Gas chromatography/negative chemical ionization mass spectrometry was employed for high sensitivity detection of the following isotopes: deuterium-labeled-linolenate, carbon-13-U-labeled-eicosapentaenoate, carbon-13-U-labeled-linoleate, and deuterium-labeled-dihomo-gamma-linolenate. These isotope-labeled fatty acids in vehicle oil were given to rats either singly or together as a single oral dose. Rat blood was collected after dosing and the isotopomers of the precursors and their main metabolites, including those containing both C-13 and H-2, were detected simultaneously with good resolution and without interference from other isotopes due to differences in mass and chromatographic retention. Published by Elsevier Science Ltd. C1 NIAAA, Sect Nutr Neurosci, Lab Membrane Biochem & Biophys, NIH, Rockville, MD 20852 USA. RP Salem, N (reprint author), NIAAA, Sect Nutr Neurosci, Lab Membrane Biochem & Biophys, NIH, 12420 Parklawn Dr,Pk Bldg,Rm 1-14, Rockville, MD 20852 USA. NR 23 TC 6 Z9 6 U1 1 U2 2 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0952-3278 J9 PROSTAG LEUKOTR ESS JI Prostaglandins Leukot. Essent. Fatty Acids PD AUG-SEP PY 2002 VL 67 IS 2-3 BP 141 EP 146 DI 10.1054/plef.411 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Cell Biology; Endocrinology & Metabolism GA 598ZW UT WOS:000178308600013 PM 12324233 ER PT J AU Loftin, CD Tiano, HF Langenbach, R AF Loftin, CD Tiano, HF Langenbach, R TI Phenotypes of the COX-deficient mice indicate physiological and pathophysiological roles for COX-1 and COX-2 SO PROSTAGLANDINS & OTHER LIPID MEDIATORS LA English DT Article DE cyclooxygenase; mice; isoform; inflammation; reproduction; cancer ID KNOCKOUT MICE; CYCLOOXYGENASE-2-DEFICIENT MICE; RENIN EXPRESSION; FEBRILE RESPONSE; GENE DISRUPTION; MUCOSAL DEFENSE; GROWTH-FACTOR; REDUCED RISK; 3T3 CELLS; INHIBITION AB The development of mice deficient in either cyclooxygenase-1 (COX-1) or COX-2, as well as mice deficient in both COX isoforms, has provided models to elucidate the physiological and pathophysiological roles of these enzymes. The findings obtained with the COX-deficient mice suggest that COX-2 may be more important than COX-1 for supplying prostaglandins (PGs) to maintain tissue homeostasis. Furthermore, both isoforms may be involved in the development of diseases, such as inflammation and cancer. Therefore, the contribution of each isoform to the prevention or development of disease is more complex than originally described. Studies with the COX-deficient mice suggest that in addition to COX-2-selective inhibition, therapeutic advances may also be achieved with COX-1-selective inhibitors which lack gastrointestinal side effects. (C) 2002 Published by Elsevier Science Inc. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, NIH, Res Triangle Pk, NC 27709 USA. RP Langenbach, R (reprint author), NIEHS, Lab Environm Carcinogenesis & Mutagenesis, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 49 TC 54 Z9 58 U1 1 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0090-6980 J9 PROSTAG OTH LIPID M JI Prostaglandins Other Lipid Mediat. PD AUG PY 2002 VL 68-9 SI SI BP 177 EP 185 AR PII S0090-6980(02)00028-X DI 10.1016/S0090-6980(02)00028-X PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 611EP UT WOS:000179003700011 PM 12432917 ER PT J AU Achanzar, WE Webber, MM Waalkes, MP AF Achanzar, WE Webber, MM Waalkes, MP TI Altered apoptotic gene expression and acquired apoptotic resistance in cadmium-transformed human prostate epithelial cells SO PROSTATE LA English DT Article DE cadmium; apoptosis; prostate; metal carcinogenesis ID NOBLE NBL/CR RAT; PROLIFERATIVE LESIONS; INJECTION SITE; CANCER RISK; IN-VITRO; CARCINOGENESIS; INDUCTION; ENVIRONMENT; MORTALITY; CASPASES AB BACKGROUND. Cadmium is a suspected prostatic carcinogen, although the underlying mechanisms are unclear. To investigate these mechanisms, we performed molecular comparisons between the cadmium-transformed prostate epithelial cell line CTPE and the nontumorigenic parental line RWPE-1. METHODS. Gene expression patterns were compared by using cDNA arrays, RNase protection assays, and Western blots. Apoptosis was analyzed by using flow cytometry to quantify apoptotic nuclei and an enzyme-linked immunosorbent assay method to measure DNA fragmentation. Caspase-3 activity was measured colorimetrically. RESULTS. Among the genes down-regulated in CTPE cells were those encoding several members of the caspase family of apoptotic proteases as well as the apoptotic regulator Bax. Ribonuclease protection assays confirmed global down-regulation of caspase gene expression in CTPE. Decreased Bax expression in CTPE was confirmed by Western blots, which also revealed increased expression of anti-apoptotic Bcl-2. Consistent with these changes, CTPE cells exhibited increased resistance to apoptosis induced by cadmium, cisplatin, and etoposide. CTPE cells also exhibited lower caspase-3 activity vs. RWPE-1 after etoposide treatment. CONCLUSIONS. CTPE cells exhibited altered expression of important apoptotic regulators as well as resistance to several apoptotic stimuli. We hypothesize that acquired apoptotic resistance may be a key aspect of cadmium-induced malignant transformation of prostate epithelial cells and that this may contribute to both tumor initiation and the acquisition of aggressive characteristics subsequent to tumor formation. (C) 2002 Wiley-Liss, Inc. C1 NIEHS, NCI, Inorgan Carcinogenesis Sect, Res Triangle Pk, NC 27709 USA. Michigan State Univ, Dept Zool, E Lansing, MI 48824 USA. Michigan State Univ, Dept Med, E Lansing, MI 48824 USA. RP Waalkes, MP (reprint author), NIEHS, NCI, Inorgan Carcinogenesis Sect, MD F0-09,111 Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 42 TC 36 Z9 40 U1 1 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0270-4137 J9 PROSTATE JI Prostate PD AUG 1 PY 2002 VL 52 IS 3 BP 236 EP 244 DI 10.1002/pros.10106 PG 9 WC Endocrinology & Metabolism; Urology & Nephrology SC Endocrinology & Metabolism; Urology & Nephrology GA 575GR UT WOS:000176939000005 PM 12111698 ER PT J AU Rigden, DJ Mosolov, VV Galperin, MY AF Rigden, DJ Mosolov, VV Galperin, MY TI Sequence conservation in the chagasin family suggests a common trend in cysteine proteinase binding by unrelated protein inhibitors SO PROTEIN SCIENCE LA English DT Article DE protease inhibitor; thiol protease; conserved domain; binding mechanism; convergent evolution ID RAY CRYSTAL-STRUCTURE; TRYPANOSOMA-CRUZI; PROTEOLYTIC-ENZYMES; CONVERGENT EVOLUTION; STRUCTURAL BASIS; PROTEASES; DISEASE; PURIFICATION; CHEMOTHERAPY; RESOLUTION AB The recently described inhibitor of cysteine proteinases from Trypanosoma cruzi, chagasin, was found to have close homologs in several eukaryotes, bacteria and archaea, the first protein inhibitors of cysteine proteases in prokaryotes. These previously uncharacterized 110-130 residue-long proteins share a well-conserved sequence motif that corresponds to two adjacent beta-strands and the short loop connecting them. Chagasin-like proteins also have other conserved, mostly aromatic, residues, and share the same predicted secondary structure. These proteins adopt an all-beta fold with eight predicted beta-strands of the immunoglobulin type. The phylogenetic distribution of the chagasins generally correlates with the presence of papain-like cysteine proteases. Previous studies have uncovered similar trends in cysteine proteinase binding by two unrelated inhibitors, stefin and p41, that belong to the cystatin and thyroglobulin families, respectively. A hypothetical model of chagasin-cruzipain interaction suggests that chagasin may dock to the cruzipain active site in a similar manner with the conserved NPTTG motif of chagasin forming a loop that is similar to the wedge structures formed at the active sites of papain and cathepsin L by stefin and p41. C1 EMBRAPA, Cenargen, Natl Ctr Genet Resources & Biotechnol, Estacao Parque Biol, BR-70770900 Brasilia, DF, Brazil. Russian Acad Sci, AN Bach Inst Biochem, Moscow 117071, Russia. NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. RP Rigden, DJ (reprint author), EMBRAPA, Cenargen, Natl Ctr Genet Resources & Biotechnol, Estacao Parque Biol, SAIN Parque Rural,Final W5, BR-70770900 Brasilia, DF, Brazil. EM daniel@cenargen.embrapa.br; vvmosolov@yahoo.com RI Galperin, Michael/B-5859-2013; OI Galperin, Michael/0000-0002-2265-5572; Rigden, Daniel/0000-0002-7565-8937 NR 56 TC 40 Z9 43 U1 0 U2 2 PU WILEY-BLACKWELL PI MALDEN PA COMMERCE PLACE, 350 MAIN ST, MALDEN 02148, MA USA SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD AUG PY 2002 VL 11 IS 8 BP 1971 EP 1977 DI 10.1110/ps.0207202 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 576ZF UT WOS:000177036500011 PM 12142451 ER PT J AU Hofmann, A Ruvinov, S Hess, S Schantz, R Delmer, DP Wlodawer, A AF Hofmann, A Ruvinov, S Hess, S Schantz, R Delmer, DP Wlodawer, A TI Plant annexins form calcium-independent oligomers in solution SO PROTEIN SCIENCE LA English DT Article DE annexin; Capsicum annuum; gel filtration; Gossypium hirsutum; sedimentation equilibrium analysis; ultracentrifugation ID PHOSPHOLIPID-BINDING PROTEINS; SELF-ASSOCIATION; MEMBRANE-BINDING; GENE-EXPRESSION; CAPSICUM-ANNUUM; CELLS; FAMILY; AGGREGATION; CALELECTRIN; SEQUENCE AB The oligomeric state in solution of four plant annexins, namely Anx23(Ca3g), Anx24(Ca32), Anx(Gh1), and Anx(Gh2), was characterized by sedimentation equilibrium analysis and gel filtration. All proteins were expressed and purified as amino-terminal His,, fusions. Sequencing of the Anx(Gh1) construct revealed distinct differences with the published sequence. Sedimentation equilibrium analysis of Anx23(Ca38), Anx24(Ca32), and Anx(Gh1) suggests monomer-trimer equilibria for each protein with association constants in the range of 0.9 x 10(10)-1.7 x 10(11) M-2. All four proteins were subjected to analytical gel filtration under different buffer conditions. Observations from this experiment series agree quantitatively with the ultracentrifugation results, and strongly suggest calcium independence of the annexin oligomerization behavior; moreover, binding of calcium ions to the proteins seems to require disassembly of the oligomers. Anx(Gh2) showed a different elution profile than the other plant annexins; while having only a very small trimer content, this annexin seems to exist in a monomer-dimer equilibrium in solution. C1 NCI, Prot Struct Sect, Macromol Crystallog Lab, Frederick, MD 21702 USA. NCI, Biochem Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. NIDDK, Struct Mass Spectrometry Facil, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. CNRS, Inst Biol Mol Plantes, F-67084 Strasbourg, France. Univ Calif Davis, Plant Biol Sect, Davis, CA 95616 USA. RP Hofmann, A (reprint author), NCI, Prot Struct Sect, Macromol Crystallog Lab, Frederick, MD 21702 USA. RI Hess, Sonja/K-4842-2013; Hofmann, Andreas/B-9515-2008 OI Hess, Sonja/0000-0002-5904-9816; Hofmann, Andreas/0000-0003-4408-5467 NR 33 TC 16 Z9 17 U1 0 U2 4 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD AUG PY 2002 VL 11 IS 8 BP 2033 EP 2040 DI 10.1110/ps.4770102 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 576ZF UT WOS:000177036500017 PM 12142457 ER PT J AU Shatsky, M Nussinov, R Wolfson, HJ AF Shatsky, M Nussinov, R Wolfson, HJ TI Flexible protein alignment and hinge detection SO PROTEINS-STRUCTURE FUNCTION AND GENETICS LA English DT Article DE structural comparison; hinge bending; flexible structural comparison; efficient algorithm; protein structural comparison ID MULTIPLE STRUCTURE ALIGNMENT; DOMAIN MOVEMENTS; DATA-BANK; SEQUENCE; IDENTIFICATION; ALGORITHMS; EFFICIENT; MOTIFS; CORE AB Here we present a novel technique for the alignment of flexible proteins. The method does not require an a priori knowledge of the flexible hinge regions. The FlexProt algorithm simultaneously detects the hinge regions and aligns the rigid subparts of the molecules. Our technique is not sensitive to insertions and deletions. Numerous methods have been developed to solve rigid structural comparisons. Unlike FlexProt, all previously developed methods designed to solve the protein flexible alignment require an a priori knowledge of the hinge regions. The FlexProt method is based on 3-D pattern-matching algorithms combined with graph theoretic techniques. The algorithm is highly efficient. For example, it performs a structural comparison of a pair of proteins with 300 amino acids in about 7 s on a 400-MHz desktop PC. We provide experimental results obtained with this algorithm. First, we flexibly align pairs of proteins taken from the database of motions. These are extended by taking additional proteins from the same SCOP family. Next, we present some of the results obtained from exhaustive all-against-all flexible structural comparisons of 1329 SCOP family representatives. Our results include relatively high-scoring flexible structural alignments between the C-terminal merozoite surface protein vs. tissue factor; class II aminoacyl-tRNA synthase, histocompatibility antigen vs. neonatal FC receptor; tyrosine-protein kinase C-SRC vs. haematopoetic cell kinase (HCK); tyrosine-protein kinase C-SRC vs. titine protein (autoinhibited serine kinase domain); and tissue factor vs. hormone-binding protein. These are illustrated and discussed, showing the capabilities of this structural alignment algorithm, which allows un-predefined hinge-based motions. (C) 2002 Wiley-Liss, Inc. C1 NCI, NCI Frederick, Intramural Res Support Program, SAIC,Lab Expt & Computat Biol, Frederick, MD 21702 USA. Tel Aviv Univ, Sch Comp Sci, IL-69978 Tel Aviv, Israel. Tel Aviv Univ, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, Israel. Tel Aviv Univ, Sackler Fac Med, Dept Human Genet & Mol Med, IL-69978 Tel Aviv, Israel. RP Nussinov, R (reprint author), NCI, NCI Frederick, Intramural Res Support Program, SAIC,Lab Expt & Computat Biol, Bldg 469,Room 151, Frederick, MD 21702 USA. RI Wolfson, Haim/A-1837-2011 FU NCI NIH HHS [N01-CO-56000] NR 42 TC 95 Z9 101 U1 0 U2 6 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-3585 J9 PROTEINS JI Proteins PD AUG 1 PY 2002 VL 48 IS 2 BP 242 EP 256 DI 10.1002/prot.10100 PG 15 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 570EN UT WOS:000176646100012 PM 12112693 ER PT J AU Kessler, RC Andrews, G Colpe, LJ Hiripi, E Mroczek, DK Normand, SLT Walters, EE Zaslavsky, AM AF Kessler, RC Andrews, G Colpe, LJ Hiripi, E Mroczek, DK Normand, SLT Walters, EE Zaslavsky, AM TI Short screening scales to monitor population prevalences and trends in non-specific psychological distress SO PSYCHOLOGICAL MEDICINE LA English DT Article ID INTERNATIONAL DIAGNOSTIC INTERVIEW; PSYCHIATRIC-DISORDERS; VALIDITY; COMORBIDITY; RELIABILITY; DEPRESSION; VALIDATION; INSTRUMENT; HISTORY AB Background. A 10-question screening scale of psychological distress and a six-question short-form scale embedded within the 10-question scale were developed for the redesigned US National Health Interview Survey (NHIS). Methods. Initial pilot questions were administered in a US national mail survey (N = 1401). A reduced set of questions was subsequently administered in a US national telephone survey (N = 1574). The 10-question and six-question scales, which we refer to as the K10 and K6, were constructed from the reduced set of questions based on Item Response Theory models. The scales were subsequently validated in a two-stage clinical reappraisal survey (N = 1000 telephone screening interviews in the first stage followed by N = 153 face-to-face clinical interviews in the second stage that oversampled first-stage respondents who screened positive for emotional problems) in a local convenience sample. The second-stage sample was administered the screening scales along with the Structured Clinical Interview for DSM-IV (SCID). The K6 was subsequently included in the 1997 (N = 36116) and 1998 (N = 32440) US National Health Interview Survey, while the K10 was included in the 1997 (N = 10641) Australian National Survey of Mental Health and Well-Being. Results. Both the K10 and K6 have good precision in the 90th-99th percentile range of the population distribution (standard errors of standardized scores in the range 0(.)20-0(.)25) as well as consistent psychometric properties across major sociodemographic subsamples. The scales strongly discriminate between community cases and non-cases of DSM-IV/SCID disorders, with areas under the Receiver Operating Characteristic (ROC) curve of 0(.)87-0(.)88 for disorders having Global Assessment of Functioning (GAF) scores of 0-70 and 0(.)95-0(.)96 for disorders having GAF scores of 0-50. Conclusions. The brevity, strong psychometric properties, and ability to discriminate DSM-IV cases from non-cases make the K10 and K6 attractive for use in general-purpose health surveys. The scales are already being used in annual government health surveys in the US and Canada as well as in the WHO World Mental Health Surveys. Routine inclusion of either the K10 or K6 in clinical studies would create an important, and heretofore missing, crosswalk between community and clinical epidemiology. C1 Harvard Univ, Sch Med, Dept Hlth Care Policy, Boston, MA 02115 USA. NIMH, Div Mental Disorders Behav Res & AIDS, Bronx, NY 10458 USA. Harvard Univ, Sch Publ Hlth, Dept Biostat, Boston, MA 02115 USA. Univ New S Wales, Clin Res Unit Anxiety Disorders, Sch Psychiat, Kensington, NSW 2033, Australia. Fordham Univ, Dept Psychol, Bronx, NY 10458 USA. RP Kessler, RC (reprint author), Harvard Univ, Sch Med, Dept Hlth Care Policy, 180 Longwood Ave, Boston, MA 02115 USA. RI Andrews, Gavin/J-6537-2012; Page, Andrew/G-5438-2012 OI Andrews, Gavin/0000-0002-4315-2173; Page, Andrew/0000-0003-3133-2844 NR 57 TC 2239 Z9 2264 U1 26 U2 172 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH ST, NEW YORK, NY 10011-4221 USA SN 0033-2917 J9 PSYCHOL MED JI Psychol. Med. PD AUG PY 2002 VL 32 IS 6 BP 959 EP 976 DI 10.1017/S0033291702006074 PG 18 WC Psychology, Clinical; Psychiatry; Psychology SC Psychology; Psychiatry GA 586TE UT WOS:000177600500002 PM 12214795 ER PT J AU Arnold, MC Papanicolaou, DA O'Grady, JA Lotsikas, A Dale, JK Straus, SE Grafman, J AF Arnold, MC Papanicolaou, DA O'Grady, JA Lotsikas, A Dale, JK Straus, SE Grafman, J TI Using an interleukin-6 challenge to evaluate neuropsychological performance in chronic fatigue syndrome SO PSYCHOLOGICAL MEDICINE LA English DT Article ID RANDOMIZED CONTROLLED-TRIAL; MULTIPLE-SCLEROSIS; DEPRESSION; DEFICITS; DISEASE; HUMANS; MEMORY; DEFINITION; IMPAIRMENT; SECRETION AB Background. Individuals with acute infections experience a range of symptoms including fatigue, malaise, muscle aches, and difficulties with concentration and memory that are usually self-limited. This cluster of symptoms is otherwise, similar to those that characterize chronic fatigue syndrome (CFS). The goal of the present study was to evaluate the cognitive and psychological functioning of CFS patients and normal controls (NCs) when they both were experiencing acute influenza-like symptoms. To induce influenza-like symptoms, we administered interleukin-6 (IL-6), a cytokine that temporarily activates the acute phase immunological and endocrine responses. Methods. Nineteen patients who met the 1994 International CFS Study Group Criteria and ten normal controls (NCs) completed routine clinical evaluations, neuropsychological tests of short-term memory, selective attention, and executive control, and self-ratings of somatic symptoms and psychological mood before, shortly following, and 1 day after IL-6 administration. Results. CFS patients consistently reported more somatic symptoms, even when both groups perceived that they were ill. Both groups somatic symptoms increased during the IL-6 challenge, but the CFS patients symptoms increased more rapidly than controls. In general, the CFS patients performed similarly to NCs on the cognitive measures before, during, and after the IL-6. In contrast to predictions, IL-6 provocation did not impair the cognitive performance of either CFS patients or NCs. Conclusions. The IL-6 provocation exacerbated the patients self-reported symptoms but did not reveal notable cognitive impairments between patients and controls during cytokine-induced acute influenza-like symptoms. C1 NINDS, NIH, Cognit Neurosci Sect, Bethesda, MD 20892 USA. NICHHD, NIH, Bethesda, MD USA. NIAID, NIH, Bethesda, MD USA. Emory Univ, Dept Med, Atlanta, GA 30322 USA. RP Grafman, J (reprint author), NINDS, NIH, Cognit Neurosci Sect, Bldg 10,Room 5C205,10 Ctr Dr,MSC 1440, Bethesda, MD 20892 USA. OI Grafman, Jordan H./0000-0001-8645-4457 NR 52 TC 18 Z9 18 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 32 AVENUE OF THE AMERICAS, NEW YORK, NY 10013-2473 USA SN 0033-2917 J9 PSYCHOL MED JI Psychol. Med. PD AUG PY 2002 VL 32 IS 6 BP 1075 EP 1089 DI 10.1017/S0033291702006086 PG 15 WC Psychology, Clinical; Psychiatry; Psychology SC Psychology; Psychiatry GA 586TE UT WOS:000177600500012 PM 12214788 ER PT J AU Chausmer, AL Elmer, GI Rubinstein, M Low, MJ Grandy, DK Katz, JL AF Chausmer, AL Elmer, GI Rubinstein, M Low, MJ Grandy, DK Katz, JL TI Cocaine-induced locomotor activity and cocaine discrimination in dopamine D2 receptor mutant mice SO PSYCHOPHARMACOLOGY LA English DT Article DE dopamine; D2; knockout; cocaine; raclopride; locomotor activity; drug discrimination ID SQUIRREL-MONKEYS; DEFICIENT MICE; STIMULUS; RATS; AGONISTS; SUBTYPES; D-2; D1 AB Rationale: Dopamine (DA) D2-like antagonists block several effects of cocaine, including its locomotor stimulant and interoceptive discriminative-stimulus effects. Because these compounds generally lack selectivity among the D2-like DA receptors, the specific roles of the subtypes remain unclear. Objectives: DA D2 receptor knockout (DA D2R KO), heterozygous (HET), and wildtype (WT) mice were used to study the role of D2 DA receptors in the effects of cocaine. Some effects of the relatively selective DA D2-like antagonist raclopride were also studied to further assess the role of D2 receptors. Methods: DA D2R KO, HET, and WT mice were treated with cocaine (1-10 mg/kg) or vehicle, and their horizontal locomotor activity was assessed. The mice were also trained to discriminate i.p. injections of saline from cocaine (10 mg/kg) using a two-response key, fixed-ratio-20 response, food-reinforcement procedure. A range of doses of cocaine (1.0-17 mg/kg) was administered before 15-min test sessions. Results: Both DA D2R KO and HET mice showed reduced levels of horizontal activity relative to WT mice. Cocaine dose dependently stimulated activity in each genotype, with the highest level of activity induced in the DA D2R WT mice. All three genotypes acquired the discrimination of 10 mg/kg cocaine; tested doses of 1.0-10.0 mg/kg produced dose-related increases in the number of cocaine-appropriate responses. Raclopride, at inactive to fully active doses (0.1-1.0 mg/kg), did not fully substitute for cocaine. Raclopride dose dependently shifted the cocaine dose-effect curve to the right in DA D2R WT and HET mice. However, in DA D2R KO mice, raclopride was inactive as an antagonist. Conclusions: The present data indicate an involvement of D2 DA receptors in the locomotor-stimulating effects and the interoceptive discriminative-stimulus effects of cocaine in WT subjects. However, the D2 receptor is not necessary for the effects, suggesting redundant dopaminergic mechanisms for the discriminative-stimulus interoceptive effects of cocaine. C1 NIDA, Psychobiol Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Univ Maryland, Sch Med, Dept Psychiat, Maryland Psychiat Res Ctr, Baltimore, MD 21201 USA. Univ Maryland, Sch Med, Dept Pharmacol & Expt Therapeut, Maryland Psychiat Res Ctr, Baltimore, MD 21201 USA. Oregon Hlth & Sci Univ, Dept Physiol & Pharmacol, Portland, OR 97201 USA. RP Katz, JL (reprint author), NIDA, Psychobiol Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. EM jkatz@intra.nida.nih.gov OI Katz, Jonathan/0000-0002-1068-1159 FU NIDA NIH HHS [DA12062] NR 30 TC 47 Z9 47 U1 1 U2 1 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0033-3158 EI 1432-2072 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD AUG PY 2002 VL 163 IS 1 BP 54 EP 61 DI 10.1007/s00213-002-1142-y PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 591JK UT WOS:000177876100008 PM 12185400 ER PT J AU Gasior, M Jaszyna, M Munzar, P Witkin, JM Goldberg, SR AF Gasior, M Jaszyna, M Munzar, P Witkin, JM Goldberg, SR TI Caffeine potentiates the discriminative-stimulus effects of nicotine in rats SO PSYCHOPHARMACOLOGY LA English DT Article DE caffeine; nicotine; drug discrimination; rat ID LOCOMOTOR-ACTIVITY; PHYSICAL-DEPENDENCE; INDUCED STIMULATION; INTRAVENOUS NICOTINE; TOBACCO WITHDRAWAL; CROSS-TOLERANCE; COCAINE; DOPAMINE; ALCOHOL; COFFEE AB Rationale: Caffeine and nicotine are the main psychoactive ingredients of coffee and tobacco, respectively, with a high frequency of concurrent use in humans. Objectives: The aim of the present study was to examine the interaction of caffeine and nicotine in rats trained to discriminate nicotine from saline. Methods: Two groups of male Sprague-Dawley rats (n=8 per group) were trained to discriminate 0.4 mg/kg nicotine, SC, from saline under a fixed ratio schedule of food presentation. One group of rats was chronically exposed to caffeine (1.0 mg/ml) dissolved in their drinking water whereas the other group was exposed to tap water. Effects of IP injections of caffeine on nicotine-lever selection were subsequently examined. In separate groups of rats exposed to the same caffeine-drinking or water-drinking regimen, effects of caffeine pretreatment on nicotine plasma levels were evaluated. Results: Although caffeine (1.0-30.0 mg/kg) did not generalize to nicotine when administered alone, it markedly potentiated discriminative-stimulus effects of the threshold dose of nicotine (0.05 mg/kg) in both water- and caffeine-drinking rats. Nicotine plasma levels were, however, not affected by acute or chronic caffeine exposure. Conclusions: Caffeine appears to enhance the discriminative-stimulus effects of the threshold dose of nicotine by a pharmacodynamic rather than a pharmacokinetic interaction. This suggests that caffeine consumption may be a contributing factor in the onset, maintenance of and relapse to tobacco dependence. C1 NIDA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Harvard Univ, McLean Hosp, Sch Med,Alcohol & Drug Abuse Res Ctr, Behav Phrmacol Program,Dept Psychiat, Belmont, MA 02478 USA. RP Gasior, M (reprint author), NIDA, Intramural Res Program, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 74 TC 32 Z9 33 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0033-3158 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD AUG PY 2002 VL 162 IS 4 BP 385 EP 395 DI 10.1007/s00213-002-1113-3 PG 11 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 588DR UT WOS:000177686200007 PM 12172692 ER PT J AU Levy-Clarke, G Byrnes, GG Buggage, RR Chan, CC AF Levy-Clarke, G Byrnes, GG Buggage, RR Chan, CC TI Primary intraocular lymphoma diagnosed by fine needle aspiration biopsy of a subretinal lesion - Reply SO RETINA-THE JOURNAL OF RETINAL AND VITREOUS DISEASES LA English DT Letter C1 Sharp Rees Stealy Med Ctr, San Diego, CA USA. NEI, Bethesda, MD 20892 USA. RP Levy-Clarke, G (reprint author), Sharp Rees Stealy Med Ctr, San Diego, CA USA. NR 4 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0275-004X J9 RETINA-J RET VIT DIS JI Retin.-J. Retin. Vitr. Dis. PD AUG PY 2002 VL 22 IS 4 BP 513 EP 513 DI 10.1097/00006982-200208000-00026 PG 1 WC Ophthalmology SC Ophthalmology GA 583XT UT WOS:000177437200026 ER PT J AU Elci, OC Akpinar-Elci, M Blair, A Dosemeci, M AF Elci, OC Akpinar-Elci, M Blair, A Dosemeci, M TI Occupational dust exposure and the risk of laryngeal cancer in Turkey SO SCANDINAVIAN JOURNAL OF WORK ENVIRONMENT & HEALTH LA English DT Article DE case-referent study; case-control study; cotton; epidemiology; larynx; work; silica ID UPPER RESPIRATORY-TRACT; EPIDEMIOLOGIC EVIDENCE; ASBESTOS EXPOSURE; WOOD DUST; MORTALITY; WORKERS; CHRYSOTILE; TOBACCO; ALCOHOL; MATRIX AB Objectives A hospital-based case-referent study was conducted to identify occupational risk factors for laryngeal cancer. In a previous report an association was found between laryngeal cancer and occupations with potential dust exposure; a job-exposure matrix was developed to aid further evaluation of laryngeal cancer risks from five occupational dust exposures. Methods Among 7631 cancer cases from the Okmeydani Hospital, Istanbul, between 1979 and 1984, 958 larynx cancer cases were identified among men. After exclusions, 940 laryngeal cancer cases and 1519 referents were available. A standardized questionnaire was used to obtain basic information on the patients. Seven-digit standard occupational and industrial codes were created to classify the job and industrial titles. A job-exposure matrix was developed for occupational dusts. including silica, asbestos, wood, cotton, and grain, and age-, smoking-, and alcohol-adjusted odds ratios (OR) and 95% confidence intervals (95% CI) were calculated to evaluate risks of laryngeal cancer. Results An excess of laryngeal cancer occurred for workers potentially exposed to silica and cotton dust, particularly for supraglottic cancer (OR 1.8, 95% CI 1.3-2.3, for silica and OR 1.6, 95% CI 1.1-2.5, for cotton dust), and there was a significant dose-response relationship with silica exposure, No relationship was found between laryngeal cancer and asbestos, grain, or wood dust exposures. Conclusion Laryngeal cancer, especially supraglottic tumors, is associated with silica and cotton dust exposures in Turkey. C1 NCI, Div Canc Epidemiol & Genet, Rockville, MD USA. Izmir Chest Dis & Surg Training Hosp, Dept Resp Med, Izmir, Turkey. RP Elci, OC (reprint author), NIOSH, Div Resp Dis Studies, Field Studies Branch, MS2800,1095 Willowdale Rd, Morgantown, WV 26505 USA. NR 46 TC 17 Z9 19 U1 1 U2 3 PU SCAND J WORK ENV HEALTH PI HELSINKI PA TOPELIUKSENKATU 41A, SF-00250 HELSINKI, FINLAND SN 0355-3140 J9 SCAND J WORK ENV HEA JI Scand. J. Work Environ. Health PD AUG PY 2002 VL 28 IS 4 BP 278 EP 284 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 584XB UT WOS:000177493100010 PM 12199430 ER PT J AU Cheson, BD AF Cheson, BD TI Hematologic malignancies: New developments and future treatments SO SEMINARS IN ONCOLOGY LA English DT Review ID NON-HODGKINS-LYMPHOMA; ANTI-CD20 MONOCLONAL-ANTIBODY; CHRONIC LYMPHOCYTIC-LEUKEMIA; MULTICENTER PHASE-II; REFRACTORY LOW-GRADE; B-CELL LYMPHOMA; ACUTE PROMYELOCYTIC LEUKEMIA; ADVANCED FOLLICULAR LYMPHOMA; APOPTOTIC DEATH ACTIVATION; IODINE I-131 TOSITUMOMAB C1 Clin Invest Branch, NCI, Bethesda, MD USA. RP Cheson, BD (reprint author), Georgetown Univ Hosp, Lombardi Canc Ctr, 3800 Reservoir Rd NW, Washington, DC 20007 USA. NR 111 TC 34 Z9 35 U1 1 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0093-7754 J9 SEMIN ONCOL JI Semin. Oncol. PD AUG PY 2002 VL 29 IS 4 SU 13 BP 33 EP 45 DI 10.1053/sonc.2002.34878 PG 13 WC Oncology SC Oncology GA 584AV UT WOS:000177445800008 PM 12170431 ER PT J AU Silverberg, MJ Ahdieh, L Munoz, A Anastos, K Burk, RD Cu-Uvin, S Duerr, A Greenblatt, RM Klein, RS Massad, S Minkoff, H Muderspach, L Palefsky, J Piessens, E Schuman, P Watts, H Shah, KV AF Silverberg, MJ Ahdieh, L Munoz, A Anastos, K Burk, RD Cu-Uvin, S Duerr, A Greenblatt, RM Klein, RS Massad, S Minkoff, H Muderspach, L Palefsky, J Piessens, E Schuman, P Watts, H Shah, KV TI The impact of HIV infection and immunodeficiency on human papillomavirus type 6 or 11 infection and on genital warts SO SEXUALLY TRANSMITTED DISEASES LA English DT Article ID WOMENS INTERAGENCY HIV; HIGH-RISK; UNINFECTED WOMEN; SERONEGATIVE WOMEN; SEROPOSITIVE WOMEN; TRACT INFECTIONS; VIRUS INFECTION; NEGATIVE WOMEN; CONDYLOMATA; PREVALENCE AB Background: HIV infection and associated immunodeficiency are known to alter the course of human papillomavirus (HPV)infections and of associated diseases. Goal: This study investigated the association between HIV and HPV and genital warts. Study Design: HPV testing and physical examinations were performed in two large prospective studies: the Women's Interagency HIV Study (WHIS) and the HIV Epidemiology Research Study (HERS). Statistical methods incorporating dependencies of longitudinal data were used to examine the relationship between HIV and HPV and genital warts. Results: A total of 1008 HIV-seronegative and 2930 HIV-seropositive women were enrolled in the two studies. The prevalence of HPV 6 or 11 was 5.6 times higher in HIV-seropositive women in the WIHS and 3.6 times higher in the HERS. Genital wart prevalence increased by a factor of 3.2 in the WHIS and 2.7 in the HERS in HIV-seropositive women. In the WHIS, infection with HPV type 6 or 11, in comparison with no HPV infection, was associated with odds of genital wart prevalence of 5.1 (95% CI: 2.9-8.8), 8.8 (95% CI: 6.1-12.8), and 12.8 (95% CI: 8.8-18.8) in HIV-seronegative women, HIV-seropositive women with greater than or equal to201 CD4 cells/mul, and HIV-seropositive women with less than or equal to200 CD4 cells/mul, respectively. In the HERS, infection with HPV type 6 or 11 was associated with odds of 2.7 (95% CI: 1.6-4.6), 4.9 (95 % CI: 3.2-7.7), and 5.3 (95% CI: 3.3-8.5) in these same groups. Other HPV types showed a similar dose-response relation, but of substantially lower magnitude and statistical significance. Conclusions: HIV infection and immunodeficiency synergistically modified the relation between HPV 6 or 11 infection and genital wart prevalence. C1 Johns Hopkins Univ, Sch Hyg & Publ Hlth, Baltimore, MD 21205 USA. Albert Einstein Coll Med, Bronx, NY 10467 USA. Montefiore Med Ctr, Bronx, NY 10467 USA. Brown Univ, Miriam Hosp, Providence, RI USA. Ctr Dis Control & Prevent, Atlanta, GA USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Cook Cty Hosp, Chicago, IL 60612 USA. Maimonides Hosp, Brooklyn, NY 11219 USA. Univ So Calif, Sch Med, Los Angeles, CA USA. Georgetown Univ, Med Ctr, Washington, DC 20007 USA. Wayne State Univ, Detroit, MI USA. NICHHD, Bethesda, MD 20892 USA. RP Shah, KV (reprint author), Johns Hopkins Univ, Sch Hyg & Publ Hlth, 615 N Wolfe St,Room E-2039, Baltimore, MD 21205 USA. NR 31 TC 36 Z9 44 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0148-5717 J9 SEX TRANSM DIS JI Sex. Transm. Dis. PD AUG PY 2002 VL 29 IS 8 BP 427 EP 435 DI 10.1097/00007435-200208000-00001 PG 9 WC Infectious Diseases SC Infectious Diseases GA 581XF UT WOS:000177319300001 PM 12172526 ER PT J AU Tooze, JA Grunwald, GK Jones, RH AF Tooze, JA Grunwald, GK Jones, RH TI Analysis of repeated measures data with clumping at zero SO STATISTICAL METHODS IN MEDICAL RESEARCH LA English DT Review ID LINEAR MIXED MODELS; SAMPLE SELECTION; INFLATED POISSON; MARKOV-MODELS; REGRESSION; VARIABLES AB Longitudinal or repeated measures data with clumping at zero occur in many applications in biometrics, including health policy research, epidemiology, nutrition, and meteorology. These data exhibit correlation because they are measured on the same subject over time or because subjects may be considered repeated measures within a larger unit such as a family. They present special challenges because of the extreme non-normality of the distributions involved. A model for repeated measures data with clumping at zero, using a mixed-effects mixed-distribution model with correlated random effects, is presented. The model contains components to model the probability of a nonzero value and the mean of nonzero values, allowing for repeated measurements using random effects and allowing for correlation between the two components. Methods for describing the effect of predictor variables on the probability of nonzero values, on the mean of nonzero values, and on the overall mean amount are given. This interpretation also applies to the mixed-distribution model for cross-sectional data. The proposed methods are illustrated with analyses of effects of several covariates on medical expenditures in 1996 for subjects clustered within households using data from the Medical Expenditure Panel Survey. C1 NCI, Bethesda, MD 20892 USA. Univ Colorado, Hlth Sci Ctr, Dept Prevent Med & Biometr, Denver, CO 80262 USA. RP Tooze, JA (reprint author), NCI, Execut Plaza N,Suite 3131,6130 Execut Blvd,MSC735, Bethesda, MD 20892 USA. FU NIGMS NIH HHS [GM38519] NR 22 TC 115 Z9 115 U1 2 U2 14 PU ARNOLD, HODDER HEADLINE PLC PI LONDON PA 338 EUSTON ROAD, LONDON NW1 3BH, ENGLAND SN 0962-2802 J9 STAT METHODS MED RES JI Stat. Methods Med. Res. PD AUG PY 2002 VL 11 IS 4 BP 341 EP 355 DI 10.1191/0962280202sm291ra PG 15 WC Health Care Sciences & Services; Mathematical & Computational Biology; Medical Informatics; Statistics & Probability SC Health Care Sciences & Services; Mathematical & Computational Biology; Medical Informatics; Mathematics GA 585WU UT WOS:000177548200005 PM 12197301 ER PT J AU Norris, JW Beletsky, V Nadareishvili, Z AF Norris, JW Beletsky, V Nadareishvili, Z TI "Spontaneous" cervical arterial dissection SO STROKE LA English DT Letter C1 Univ Toronto, Toronto, ON, Canada. Univ Western Ontario, London, ON N6A 3K7, Canada. NIH, Bethesda, MD 20892 USA. RP Norris, JW (reprint author), Univ Toronto, 100 Coll St, Toronto, ON, Canada. NR 4 TC 4 Z9 4 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0039-2499 J9 STROKE JI Stroke PD AUG PY 2002 VL 33 IS 8 BP 1945 EP 1945 DI 10.1161/01.STR.0000023580.25308.D0 PG 1 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA 581XM UT WOS:000177320100005 PM 12154240 ER PT J AU Krieg, RC Paweletz, CP Liotta, LA Petricoin, EF AF Krieg, Rene C. Paweletz, Cloud P. Liotta, Lance A. Petricoin, Emanuel F., III TI Clinical Proteomics for Cancer Biomarker Discovery and Therapeutic Targeting SO TECHNOLOGY IN CANCER RESEARCH & TREATMENT LA English DT Article DE SELDI; 2D-PAGE; laser capture microdissection; proteomic pattern diagnostics; protein array AB As we emerge into the post-genome era, proteomics finds itself as the driving force field as we translate the nucleic acid information archive into understanding how the cell actually works and how disease processes operate. Even so, the traditionally held view of proteomics as simply cataloging and developing lists of the cellular protein repertoire of a cell are now changing, especially in the sub-discipline of clinical proteomics. The most relevant information archive to clinical applications and drug development involves the elucidation of the information flow of the cell; the "software" of protein pathway networks and circuitry. The deranged circuitry of the cell as the drug target itself as well as the effect of the drug on not just the target, but also the entire network, is what we now are striving towards. Clinical proteomics, as a new and most exciting sub-discipline of proteomics, involves the bench-to-bedside clinical application of proteomic tools. Unlike the genome, there are potentially thousands of proteomes: each cell type has its own unique proteome. Moreover, each cell type can alter its proteome depending on the unique tissue microenvironment in which it resides, giving rise to multiple permutations of a single proteome. Since there is no polymerase chain reaction equivalent to proteomics- identifying and discovering the "wiring diagram" of a human diseased cell in a biopsy specimen remains a daunting challenge. New micro-proteomic technologies are being and still need to be developed to drill down into the proteomes of clinically relevant material. Cancer, as a model disease, provides a fertile environment to study the application of proteomics at the bedside. The promise of clinical proteomics and the new technologies that are developed is that we will detect cancer earlier through discovery of biomarkers, we will discover the next generation of targets and imaging biomarkers, and we can then apply this knowledge to patient-tailored therapy. C1 [Krieg, Rene C.; Paweletz, Cloud P.; Liotta, Lance A.] NCI, FDA NCI Clin Prote Program, Pathol Lab, NIH, Bethesda, MD 20892 USA. [Petricoin, Emanuel F., III] US FDA, FDA NCI Clin Prote Program, Div Therapeut Prot, CBER, Bethesda, MD 20892 USA. RP Petricoin, EF (reprint author), US FDA, FDA NCI Clin Prote Program, Div Therapeut Prot, CBER, Bethesda, MD 20892 USA. EM petricoin@cber.fda.gov NR 61 TC 25 Z9 27 U1 0 U2 1 PU ADENINE PRESS PI SCHENECTADY PA 2066 CENTRAL AVE, SCHENECTADY, NY 12304 USA SN 1533-0346 J9 TECHNOL CANCER RES T JI Technol. Cancer Res. Treat. PD AUG PY 2002 VL 1 IS 4 BP 263 EP 272 PG 10 WC Oncology SC Oncology GA V28FX UT WOS:000208667800007 PM 12625785 ER PT J AU Hung, W Sarlis, NJ AF Hung, W Sarlis, NJ TI Current controversies in the management of pediatric patients with well-differentiated nonmedullary thyroid cancer: A review SO THYROID LA English DT Review ID RECOMBINANT HUMAN THYROTROPIN; POSITRON-EMISSION-TOMOGRAPHY; SERUM THYROGLOBULIN LEVELS; NEEDLE ASPIRATION BIOPSY; LOW-IODINE DIET; SURGICAL DISCIPLINE COMMITTEE; RET PROTOONCOGENE MUTATIONS; NEGATIVE RADIOIODINE SCANS; ENDOCRINE NEOPLASIA TYPE-2; TERM FOLLOW-UP AB Current treatment strategies for pediatric patients with nonmedullary, well-differentiated thyroid carcinoma (WDTC) are derived from single-institution clinical cohorts, reports of extensive personal experience, and extrapolation of several common therapeutic practices for this tumor in adults. Because pediatric WDTC is an uncommon malignancy, the issues of its optimal initial and subsequent long-term treatment and follow-up remain controversial. Pediatric patients with WDTC can be divided into two groups: children younger than 10 years of age and teenagers/adolescents between 10 and 18 years of age because these groups have different recurrence and mortality rates. We hereby present our views and interpret them in the light of the pertinent literature. Our recommendations on treatment strategies are more relevant for younger children. After midpuberty, optimal treatment is adequately addressed in the relevant literature on adults. For the majority of patients, total/near-total thyroidectomy is currently recommended as the standard initial therapy for WDTC. This is commonly followed by administration of radioiodine (RAI; I-131) therapy to destroy residual normal thyroid tissue (remnant). Routine I-131 remnant ablation has been shown to: (1). decrease the risk of local recurrences, (2) increase the sensitivity of subsequent diagnostic RAI whole-body scanning (WBS), and (3) render serum thyroglobulin (Tg) a highly sensitive marker for recurrent/residual disease during long-term follow-up. We recognize that the above practices are not universally adhered to in children and adolescents, because the risk stratification and intensity of applied therapeutic measures are influenced by institutional traditions and personal experience. In our view, aggressive initial management, followed by evaluations at regular intervals after thyroidectomy and I-131 remnant ablation, in conjunction with long-term thyroid hormone suppressive therapy (THST), result in decreased recurrence rates in pediatric patients with WDTC. Follow-up examinations should include a diagnostic RAI (I-131 or I-123) WBS and measurement of serum Tg, both performed under conditions of TSH stimulation, as well as neck ultrasonography (US). Our strategy is corroborated by data from retrospective clinical cohort studies. In this malignancy, no evidence of disease (NED) status can be defined as the combination of a negative diagnostic WBS and the presence of undetectable or low serum Tg levels, both tested under TSH stimulation. These findings should be accompanied by the absence of anatomically definable disease by standard imaging modalities, e.g., neck US or chest computed tomography (CT). Although the long-term survival rates are good overall in this disease, selected patients may require further surgery or 1311 therapy for the eradication or clinical control of metastases. Finally, and importantly, because the duration of follow-up is lifelong, the care of children with prior diagnosis of WDTC should be transferred to an adult endocrinologist after they reach adulthood, even if they have achieved NED status by that time. C1 NIDDK, DIR, NIH, Dev Endocrinol Branch, Bethesda, MD 20892 USA. Georgetown Univ, Med Ctr, Div Pediat Endocrinol & Metab, Washington, DC 20007 USA. NIDDKD, Div Intramural Res, Bethesda, MD USA. RP Sarlis, NJ (reprint author), NIDDK, DIR, NIH, Dev Endocrinol Branch, Bldg 10,Room 8D12C,10 Ctr Dr,MSC 1758, Bethesda, MD 20892 USA. NR 197 TC 79 Z9 86 U1 0 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1050-7256 J9 THYROID JI Thyroid PD AUG PY 2002 VL 12 IS 8 BP 683 EP 702 DI 10.1089/105072502760258668 PG 20 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 586BB UT WOS:000177561300006 PM 12225637 ER PT J AU Aframian, DJ Redman, RS Yamano, S Nikolovski, J Cukierman, E Yamada, KM Kriete, MF Swaim, WD Mooney, DJ Baum, BJ AF Aframian, DJ Redman, RS Yamano, S Nikolovski, J Cukierman, E Yamada, KM Kriete, MF Swaim, WD Mooney, DJ Baum, BJ TI Tissue compatibility of two biodegradable tubular scaffolds implanted adjacent to skin or buccal mucosa in mice SO TISSUE ENGINEERING LA English DT Article ID IN-VIVO BIOCOMPATIBILITY; ORAL IMMUNIZATION; SALIVARY-GLANDS; CELLS; COPOLYMERS; MODEL; MICROPARTICLES; MICROSPHERES; DEGRADATION; DYSFUNCTION AB Radiation therapy for cancer in the head and neck region leads to a marked loss of salivary gland parenchyma, resulting in a severe reduction of salivary secretions. Currently, there is no satisfactory treatment for these patients. To address this problem, we are using both tissue engineering and gene transfer principles to develop an orally implantable, artificial fluid-secreting device. In the present study, we examined the tissue compatibility of two biodegradable substrata potentially useful in fabricating such a device. We implanted in Balb/c mice tubular scaffolds of poly-L-lactic acid (PLLA), poly-glycolic acid coated with PLLA (PGA/PLLA), or nothing (sham-operated controls) either beneath the skin on the back, a site widely used in earlier toxicity and biocompatibility studies, or adjacent to the buccal mucosa, a site quite different functionally and immunologically. At 1, 3, 7, 14, and 28 days postimplantation, implant sites were examined histologically, and systemic responses were assessed by conventional clinical chemistry and hematology analyses. Inflammatory responses in the connective tissue were similar regardless of site or type of polymer implant used. However, inflammatory reactions were shorter and without epithelioid and giant cells in sham-operated controls. Also, biodegradation proceeded more slowly with the PLLA tubules than with the PGA/PLLA tubules. No significant changes in clinical chemistry and hematology were seen due to the implantation of tubular scaffolds. These results indicate that the tissue responses to PLLA and PGA/PLLA scaffolds are generally similar in areas subjacent to skin in the back and oral cavity. However, these studies also identified several potentially significant concerns that must be addressed prior to initiating any clinical applications of this device. C1 NIDCR, GTTB, NIH, Bethesda, MD 20892 USA. NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD USA. NIDCR, Anim Care Unit, NIH, Bethesda, MD USA. NIDCR, Cellular Imaging Core Facil, NIH, Bethesda, MD USA. Dept Vet Affairs Med Ctr, Oral Pathol Res Lab, Washington, DC USA. Univ Michigan, Dept Biomed Engn, Ann Arbor, MI 48109 USA. RP Baum, BJ (reprint author), NIDCR, GTTB, NIH, Bldg 10,Rm 1N113,MSC-1190, Bethesda, MD 20892 USA. OI Yamada, Kenneth/0000-0003-1512-6805; Yamano, Seiichi/0000-0003-2056-4359 NR 51 TC 26 Z9 28 U1 0 U2 8 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1076-3279 J9 TISSUE ENG JI Tissue Eng. PD AUG PY 2002 VL 8 IS 4 BP 649 EP 659 DI 10.1089/107632702760240562 PG 11 WC Cell & Tissue Engineering SC Cell Biology GA 584MR UT WOS:000177470800012 PM 12202004 ER PT J AU Wyde, ME Cambre, T Lebetkin, M Eldridge, SR Walker, NJ AF Wyde, ME Cambre, T Lebetkin, M Eldridge, SR Walker, NJ TI Promotion of altered hepatic foci by 2,3,7,8-tetrachlorodibenzo-p-dioxin and 17 beta-estradiol in male Sprague-Dawley rats SO TOXICOLOGICAL SCIENCES LA English DT Article DE carcinogenesis; dioxin; estrogen; promotion cell proliferation; oxidative damage ID DOSE-RESPONSE RELATIONSHIPS; PARA-DIOXIN-CONGENERS; OXIDATIVE DNA-DAMAGE; CELL-PROLIFERATION; CHRONIC EXPOSURE; TUMOR PROMOTION; POLYCHLORINATED-BIPHENYLS; DIBENZOFURAN-CONGENERS; OVARIECTOMIZED RATS; CATECHOL ESTROGENS AB The determination of differences in hormonal regulation of tumor promotion-related response to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) between males and females may identify factors contributing to the female-specific hepatocarcinogenicity of TCDD in rats. In the current study, diethylnitrosamine-initiated male Sprague-Dawley rats were exposed to TCDD or corn oil vehicle in the presence and absence of 17beta-estradiol (E2), and cell proliferation and development of preneoplastic altered hepatic foci (AHF) were determined. After 30 weeks of exposure, gamma-glutamyltranspeptidase (GGT)-positive AHF and the number of placental glutathione-s-transferase (PGST)-positive AHF were significantly higher in TCDD-treated rats than in control rats. Both the number and volume fraction of GGT-positive AHF were significantly lower in rats cotreated with E2 regardless of TCDD exposure compared with corresponding non-E2-treated groups and were unaffected by TCDD. In contrast, the number of PGST-positive AHF was significantly higher in E2-treated rats in the absence of TCDD treatment. In addition, whereas E2 had no effect on the volume fraction of PGST-positive foci, the levels in rats cotreated with both E2 and TCDD were significantly higher than in controls. No differences were observed in cell proliferation between TCDD-treated and control rats, although cell proliferation was lower in rats exposed to E2 compared with placebo controls. The weaker potency of tumor promotion and lack of induction of cell replication and DNA damage in male rats likely explain the female-specific hepatocarcinogenicity of TCDD in chronic bioassays. C1 NIEHS, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Curriculum Toxicol, Chapel Hill, NC 27599 USA. Pathol Associates Inc, Frederick, MD 21701 USA. RP Walker, NJ (reprint author), NIEHS, Environm Toxicol Program, 111 Alexander Dr,Bldg 101,Room D452,MD4-01, Res Triangle Pk, NC 27709 USA. EM walker3@niehs.nih.gov RI Walker, Nigel/D-6583-2012 OI Walker, Nigel/0000-0002-9111-6855 NR 58 TC 9 Z9 9 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 EI 1096-0929 J9 TOXICOL SCI JI Toxicol. Sci. PD AUG PY 2002 VL 68 IS 2 BP 295 EP 303 DI 10.1093/toxsci/68.2.295 PG 9 WC Toxicology SC Toxicology GA 580GC UT WOS:000177226000007 PM 12151625 ER PT J AU Tinwell, H Haseman, J Lefevre, PA Wallis, N Ashby, J AF Tinwell, H Haseman, J Lefevre, PA Wallis, N Ashby, J TI Normal sexual development of two strains of rat exposed in utero to low doses of bisphenol A SO TOXICOLOGICAL SCIENCES LA English DT Article DE bisphenol A; Sprague-Dawley rats; Alderley Park rats; sexual development; in utero exposure ID ESTROGENS; SPERMATOGENESIS; ASSAY AB Pregnant Sprague-Dawley (SD) and Alderley Park (Wistar derived) rats were exposed by gavage during gestation days 6-21 to 20 mug/kg, 100 mug/kg, or 50 mg/kg body weight of BPA with ethinylestradiol (EE; 200 mug/kg) acting as a positive control agent. The sexual development of the derived pups was monitored until termination at postnatal day 90-98. The endpoints evaluated were litter size and weight, anogenital distance at birth, days of vaginal opening, first estrus and prepuce separation, weights of the liver, seminal vesicles, epididimydes, testes, ventral prostate, uterus, vagina, cervix and ovaries, and daily sperm production. Males were terminated at postnatal day 90 and females at postnatal day 98. The only statistically significant effects observed for any dose of BPA were a decrease in daily sperm production and an increase in the age of vaginal opening for the Alderley Park animals at the highest dose evaluated (50 mg/kg). The dose of EE evaluated proved to be maternally toxic in our laboratory, but provided gross evidence of endocrine disruption in the treated dams. These results diverge from those of Chahoud and his colleagues who indicated disturbances to the sexual development of both male and female SD rat pups administered the same 3 doses of BPA. This failure to confirm low dose endocrine effects for BPA is discussed within the context of similar divergent conclusions derived from other assessments of the endocrine toxicity of this agent to rats. C1 Syngenta Cent Toxicol Lab, Macclesfield SK10 4TJ, Cheshire, England. NIEHS, Res Triangle Pk, NC 27709 USA. RP Ashby, J (reprint author), Syngenta Cent Toxicol Lab, Alderley Pk, Macclesfield SK10 4TJ, Cheshire, England. NR 32 TC 66 Z9 66 U1 0 U2 8 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 J9 TOXICOL SCI JI Toxicol. Sci. PD AUG PY 2002 VL 68 IS 2 BP 339 EP 348 DI 10.1093/toxsci/68.2.339 PG 10 WC Toxicology SC Toxicology GA 580GC UT WOS:000177226000012 PM 12151630 ER PT J AU Gunther, MR Sturgeon, BE Mason, RP AF Gunther, MR Sturgeon, BE Mason, RP TI Nitric oxide trapping of the tyrosyl radical-chemistry and biochemistry SO TOXICOLOGY LA English DT Article DE nitric oxide; vasodilation; tyrosyl radical-chemistry ID PROSTAGLANDIN-H SYNTHASE; RIBONUCLEOTIDE REDUCTASE; PHOTOSYSTEM-II; ESCHERICHIA-COLI; ENDOPEROXIDE SYNTHASE; ANACYSTIS-NIDULANS; HYDROGEN-PEROXIDE; ELECTRON-TRANSFER; DNA PHOTOLYASE; COMPOUND-I AB The quenching of the Y-D tyrosyl radical in photosystem II by nitric oxide was reported to result from the formation of a weak tyrosyl radical-nitric oxide complex. This radical/radical reaction is expected to generate an electron spin resonance (ESR)-silent nitrosocyclohexadietione species that can reversibly regenerate the tyrosyl radical and nitric oxide or undergo rearrangement to form 3-nitrosotyrosine. It has been proposed that 3-nitrosotyrosine can be oxidized by one electron to form the tyrosine iminoxyl radical ( > C = N-O-.). This proposal was put forth as a result of ESR detection of the iminoxyl radical intermediate when photosystem II was exposed to nitric oxide. Although the detection of the iminoxyl radical in photosystem II strongly suggested a mechanism involving 3-nitrosotyrosine, the iminoxyl radical ESR spectrum was not unequivocally identified as originating from tyrosine. Subsequently, non-protein L-tyrosine iminoxyl radical was generated by two methods: (1) peroxidase oxidation of synthetic 3-nitroso-N-acetyl-L-tyrosine; and (2) peroxidase oxidation of free L-tyrosine in the presence of nitric oxide. The determination of protein nitrotyrosine content has become a frequently used technique for the detection of nitrosative tissue damage. Protein nitration has been suggested to be a final product of the production of highly reactive nitrogen oxide intermediates (e.g. peroxynitrite) formed in reactions between nitric oxide (NO.) and oxygen-derived species such as superoxide. The enzyme prostaglandin H synthase-2 also forms a tyrosyl radical during its enzymatic catalysis of prostaglandin formation. In the presence of the NO.-generator diethylamine nonoate, the tyrosyl radical of prostaglandin H synthase-2 also changes to that of an iminoxyl radical. Western blot analysis of prostaglandin H synthase-2 after exposure to the NO.-generator revealed nitrotyrosine formation. The results provide a mechanism for nitric oxide-dependent tyrosine nitration that does not require formation of more highly reactive nitrogen oxide intermediates such as peroxynitrite or nitrogen dioxide. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. RP Mason, RP (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 33 TC 35 Z9 36 U1 0 U2 4 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0300-483X J9 TOXICOLOGY JI Toxicology PD AUG 1 PY 2002 VL 177 IS 1 BP 1 EP 9 AR PII S0300-483X(02)00191-9 DI 10.1016/S0300-483X(02)00191-9 PG 9 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 585QW UT WOS:000177536900001 PM 12126791 ER PT J AU Taube, JR Gao, CY Ueda, Y Zelenka, PS David, LL Duncan, MK AF Taube, JR Gao, CY Ueda, Y Zelenka, PS David, LL Duncan, MK TI General utility of the chicken beta B1-Crystallin promoter to drive protein expression in lens fiber cells of transgenic mice SO TRANSGENIC RESEARCH LA English DT Article DE cataract; beta B1-crystallin; eye; gene expression; lens; transgenic mice ID ALPHA-A-CRYSTALLIN; EUKARYOTIC MESSENGER-RNAS; B-CRYSTALLIN; EYE LENS; DEVELOPMENTAL REGULATION; DELTA-1-CRYSTALLIN GENE; 3'-UNTRANSLATED REGIONS; DIFFERENTIAL EXPRESSION; PREFERRED ACTIVITY; DELTA-CRYSTALLIN AB Transgenic mouse technology has been very valuable for the study of lens fiber cells since they can not be propagated in cell culture. The targeting of transgenes to the lens has traditionally been done with the alphaA-crystallin promoter. However, while lens-specific, transgenic lines made with the alphaA-crystallin promoter express the transgene at levels 100-300-fold lower than endogenous alphaA-crystallin. Here we propose an alternative, the chicken betaB1-crystallin promoter (-432/+30). Transgenic mice made with this promoter have successfully expressed CAT, d/n m-calpain, Wee1, and betaB2-crystallin mRNA at levels comparable to the endogenous betaB1-crystallin gene and no eye abnormalities such as cataracts, have resulted. All of the transgenic lines made with the chicken betaB1-crystallin promoter have expressed the transgene in the lens fiber cells, and the best lines express at levels close to endogenous betaB1-crystallin. While RNA expression is very high, only moderate protein expression has been achieved, implying that the high protein expression of the crystallins is partially controlled at the level of translation. Thus, the chicken betaB1-crystallin promoter directs high level RNA expression to lens fiber cells, which may be especially useful for the expression of ribozyme and anti-sense RNAs in addition to ectopic proteins. C1 Univ Delaware, Dept Biol Sci, Newark, DE 19716 USA. NEI, Mol & Dev Biol Lab, Bethesda, MD 20892 USA. Oregon Hlth Sci Univ, Sch Dent, Dept Oral Mol Biol, Portland, OR 97201 USA. Oregon Hlth Sci Univ, Sch Dent, Dept Ophthalmol, Portland, OR 97201 USA. Oregon Hlth Sci Univ, Sch Med, Dept Ophthalmol, Portland, OR 97201 USA. Oregon Hlth Sci Univ, Sch Med, Dept Oral Mol Biol, Portland, OR 97201 USA. RP Duncan, MK (reprint author), Univ Delaware, Dept Biol Sci, 314 Wolf Hall, Newark, DE 19716 USA. FU NEI NIH HHS [EY12016, EY12221] NR 70 TC 9 Z9 9 U1 1 U2 2 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0962-8819 J9 TRANSGENIC RES JI Transgenic Res. PD AUG PY 2002 VL 11 IS 4 BP 397 EP 410 DI 10.1023/A:1016364001095 PG 14 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology GA 573GJ UT WOS:000176820700007 PM 12212842 ER PT J AU Makarova, KS Aravind, L Koonin, EV AF Makarova, KS Aravind, L Koonin, EV TI SWIM, a novel Zn-chelating domain present in bacteria, archaea and eukaryotes SO TRENDS IN BIOCHEMICAL SCIENCES LA English DT Article ID ZINC-FINGER PROTEINS; TRANSCRIPTION FACTORS; TRANSPOSABLE ELEMENT; SUPERFAMILY; MAIZE AB A previously undetected domain with a CxCx(n) CxH pattern of predicted zinc-chelating residues was identified in a variety of prokaryotic and eukaryotic proteins. These include bacterial ATPases of the SW12/SNF2 family, plant MuDR transposases and transposase-derived Far1 nuclear proteins, and vertebrate MEK kinase-1. This domain was designated SWIM after SW12/SNF2 and MuDR, and is predicted to have DNA-binding and protein-protein interaction functions in different contexts. C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. RP Makarova, KS (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. NR 17 TC 39 Z9 41 U1 1 U2 1 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0968-0004 J9 TRENDS BIOCHEM SCI JI Trends Biochem.Sci. PD AUG PY 2002 VL 27 IS 8 BP 384 EP 386 AR PII S0968-0004(02)02140-0 DI 10.1016/S0968-0004(02)02140-0 PG 3 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 586JV UT WOS:000177581600002 PM 12151216 ER PT J AU Uhl, GR Liu, QR Naiman, D AF Uhl, GR Liu, QR Naiman, D TI Substance abuse vulnerability loci: converging genome scanning data SO TRENDS IN GENETICS LA English DT Review ID RECEPTOR KNOCKOUT MICE; MORPHINE-INDUCED ANALGESIA; DRUG-ABUSE; ALCOHOL DEPENDENCE; ENVIRONMENTAL-INFLUENCES; FEMALE TWINS; LINKAGE; GENES; RISK; DISORDERS AB Classical genetic studies suggest strong complex genetic contributions to a predisposition to abuse multiple addictive substances. Until recently, there were no reproducible genome scanning data identifying chromosomal positions likely to contain allelic variants that predispose the carrier to illegal substance addiction. Nominal results of linkage-based genome scanning studies for ethanol and nicotine addictions failed to display much agreement. Our recent data from association-based genome scans for illegal addictions, and reanalyses of previous results now provide a substantial body of converging results. The 15 reproducible chromosomal loci identified here are good candidates to harbor allelic variants that alter human substance abuse vulnerabilities. We discuss several approaches to identifying the specific gene variants that underlie these convergent association and linkage observations, and the impact that these convergent observations should have on understanding important human addictive disorders. C1 NIDA, Mol Neurobiol Branch, IRP, NIH, Baltimore, MD 21224 USA. Johns Hopkins Univ, Dept Math Sci, Baltimore, MD 21218 USA. RP Uhl, GR (reprint author), NIDA, Mol Neurobiol Branch, IRP, NIH, Box 5180, Baltimore, MD 21224 USA. RI Naiman, Daniel/A-3304-2010; Liu, Qing-Rong/A-3059-2012 OI Naiman, Daniel/0000-0001-6504-9081; Liu, Qing-Rong/0000-0001-8477-6452 NR 49 TC 72 Z9 72 U1 5 U2 10 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0168-9525 J9 TRENDS GENET JI Trends Genet. PD AUG PY 2002 VL 18 IS 8 BP 420 EP 425 AR PII S0168-9525(02)02719-1 DI 10.1016/S0168-9525(02)02719-1 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 578HK UT WOS:000177111200008 PM 12142011 ER PT J AU Giuditta, A Kaplan, BB van Minnen, J Alvarez, J Koenig, E AF Giuditta, A Kaplan, BB van Minnen, J Alvarez, J Koenig, E TI Axonal and presynaptic protein synthesis: new insights into the biology of the neuron SO TRENDS IN NEUROSCIENCES LA English DT Editorial Material ID SQUID GIANT-AXON; LONG-TERM FACILITATION; ACTIN MESSENGER-RNA; GROWTH CONES; SYMPATHETIC NEURONS; MYELINATED AXONS; BETA-ACTIN; SYNAPTOSOMAL FRACTION; LOCAL SYNTHESIS; RIBOSOMAL-RNA AB The presence of a local mRNA translation system in axons and terminals was proposed almost 40 years ago. Over the ensuing period, an impressive body of evidence has grown to support this proposal - yet the nerve cell body is still considered to be the only source of axonal and presynaptic proteins. To dispel this lingering neglect, we now present the wealth of recent observations bearing on this central idea, and consider their impact on our understanding of the biology of the neuron. We demonstrate that extrasomatic translation sites, which are now well recognized in dendrites, are also present in axonal and presynaptic compartments. C1 Univ Naples Federico II, Dept Gen & Environm Physiol, Naples, Italy. NIMH, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Vrije Univ Amsterdam, Grad Sch Neurosci, Neurosci Res Inst, Fac Earth & Life Sci, Amsterdam, Netherlands. Pontificia Univ Catolica Chile, Dept Cellular & Mol Biol, Santiago, Chile. Univ Buffalo, Dept Physiol & Biophys, Buffalo, NY USA. RP Giuditta, A (reprint author), Univ Naples Federico II, Dept Gen & Environm Physiol, Naples, Italy. EM giuditta@unina.it FU NIGMS NIH HHS [GM39600]; NINDS NIH HHS [NS30785] NR 70 TC 130 Z9 132 U1 1 U2 8 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0166-2236 J9 TRENDS NEUROSCI JI Trends Neurosci. PD AUG PY 2002 VL 25 IS 8 BP 400 EP 404 DI 10.1016/S0166-2236(02)02188-4 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 576HU UT WOS:000176998500013 PM 12127756 ER PT J AU Masson, GS Yang, DS Miles, FA AF Masson, GS Yang, DS Miles, FA TI Reversed short-latency ocular following SO VISION RESEARCH LA English DT Article DE apparent motion; ocular tracking; reversed phi motion; motion detectors ID VERGENCE EYE-MOVEMENTS; 2ND-ORDER MOTION; OPTOKINETIC NYSTAGMUS; BINOCULAR DISPARITY; FIRST-ORDER; CONTRAST MODULATIONS; VIEWING DISTANCE; DEPTH-PERCEPTION; APPARENT MOTION; FOURIER MOTION AB Using the scleral search coil technique to monitor eye movements, we recorded short-latency ocular following responses to displacement steps of large random-dot patterns. On half of the trials, the luminance of the dots and background were reversed during the step, a procedure that is known to reverse the direction of the perceived motion ("reverse phi"). Steps without luminance reversal induced small but consistent ocular following in the direction of the steps at ultra-short latency (<80 ms). Steps with luminance reversal induced small but Consistent tracking at the same latency but in the direction opposite to the actual displacement. Tuning curves describing the dependence of initial ocular following on the amplitude of the displacement had a form approximating the derivative of a Gaussian and were well fit by Gabor functions, the cosine term being phase shifted similar to180degrees by the luminance reversal. This result is consistent with the idea that the initial ocular following is mediated, at least in part. by first-order (luminance) motion-energy detectors. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 CNRS, FRE2098, Ctr Rech Neurosci Cognit, F-13402 Marseille 20, France. NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. RP Masson, GS (reprint author), CNRS, FRE2098, Ctr Rech Neurosci Cognit, 31 Chemin Joseph Aiguier, F-13402 Marseille 20, France. RI MASSON, Guillaume/G-4615-2012 FU Intramural NIH HHS [Z01 EY000153-24] NR 47 TC 28 Z9 28 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0042-6989 J9 VISION RES JI Vision Res. PD AUG PY 2002 VL 42 IS 17 BP 2081 EP 2087 AR PII S0042-6989(02)00082-2 DI 10.1016/S0042-6989(02)00082-2 PG 7 WC Neurosciences; Ophthalmology SC Neurosciences & Neurology; Ophthalmology GA 596ZJ UT WOS:000178196800005 PM 12169427 ER PT J AU Wlodawer, A AF Wlodawer, A TI Structure-based design of AIDS drugs and the development of resistance SO VOX SANGUINIS LA English DT Article; Proceedings Paper CT 27th Congress of the international-Society-of-Blood-Transfusion CY AUG 24-29, 2002 CL VANCOUVER, CANADA SP Int Soc Blood Transfusion ID HUMAN-IMMUNODEFICIENCY-VIRUS; HIV-PROTEINASE-INHIBITORS; CRYSTAL-STRUCTURE; REVERSE-TRANSCRIPTASE; PROTEASE INHIBITOR; CATALYTIC DOMAIN; 3-DIMENSIONAL STRUCTURE; ANGSTROM RESOLUTION; TRANSITION-STATE; TYPE-2 PROTEASE C1 NCI, Macromol Crystallog Lab, Frederick, MD 21701 USA. RP Wlodawer, A (reprint author), NCI, Macromol Crystallog Lab, Frederick, MD 21701 USA. NR 46 TC 10 Z9 11 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0042-9007 J9 VOX SANG JI Vox Sang. PD AUG PY 2002 VL 83 SU 1 BP 23 EP 26 DI 10.1111/j.1423-0410.2002.tb05261.x PG 4 WC Hematology SC Hematology GA 607UZ UT WOS:000178810300008 PM 12617097 ER PT J AU Alving, BM AF Alving, BM TI Platelet substitutes: The reality and the potential SO VOX SANGUINIS LA English DT Article; Proceedings Paper CT 27th Congress of the international-Society-of-Blood-Transfusion CY AUG 24-29, 2002 CL VANCOUVER, CANADA SP Int Soc Blood Transfusion ID RECOMBINANT FACTOR VIIA; IN-VIVO; BLOOD-LOSS; THROMBOCYTOPENIA; TRANSFUSION; MODEL; FRESH C1 NHLBI, NIH, Bethesda, MD 20892 USA. RP Alving, BM (reprint author), NHLBI, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 16 TC 3 Z9 3 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0042-9007 J9 VOX SANG JI Vox Sang. PD AUG PY 2002 VL 83 SU 1 BP 287 EP 288 DI 10.1111/j.1423-0410.2002.tb05319.x PG 2 WC Hematology SC Hematology GA 607UZ UT WOS:000178810300066 PM 12617154 ER PT J AU Stroncek, D AF Stroncek, D TI Neutrophil-specific antigen HNA-2a (NB1, CD177): Serology, biochemistry, and molecular biology SO VOX SANGUINIS LA English DT Article; Proceedings Paper CT 27th Congress of the international-Society-of-Blood-Transfusion CY AUG 24-29, 2002 CL VANCOUVER, CANADA SP Int Soc Blood Transfusion ID MONOCLONAL-ANTIBODIES; TRANSFUSION REACTIONS; GENE-FREQUENCIES; GLYCOPROTEIN; EXPRESSION; NEUTROPENIAS; PATHOGENESIS C1 NIH, Dept Transfus Med, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Stroncek, D (reprint author), NIH, Dept Transfus Med, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. NR 32 TC 4 Z9 4 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0042-9007 J9 VOX SANG JI Vox Sang. PD AUG PY 2002 VL 83 SU 1 BP 359 EP 361 DI 10.1111/j.1423-0410.2002.tb05334.x PG 3 WC Hematology SC Hematology GA 607UZ UT WOS:000178810300081 PM 12617169 ER PT J AU Ohiro, Y Garkavtsev, I Kobayashi, S Sreekumar, KR Nantz, R Higashikubo, BT Duffy, SL Higashikubo, R Usheva, A Gius, D Kley, N Horikoshi, N AF Ohiro, Y Garkavtsev, I Kobayashi, S Sreekumar, KR Nantz, R Higashikubo, BT Duffy, SL Higashikubo, R Usheva, A Gius, D Kley, N Horikoshi, N TI A novel p53-inducible apoptogenic gene, PRG3, encodes a homologue of the apoptosis-inducing factor (AIF) SO FEBS LETTERS LA English DT Article DE p53; target gene; cloning; apoptosis; oxidoreductase; transcription ID WILD-TYPE P53; CELL-CYCLE ARREST; P53-DEPENDENT APOPTOSIS; POTENTIAL MEDIATOR; PROMOTES APOPTOSIS; DNA-DAMAGE; PROTEIN; DOMAIN; EXPRESSION; INHIBITOR AB The p53 tumor suppressor protein induces cell cycle arrest or apoptosis in response to cellular stresses. We have identified PRG3 (p53-responsive gene 3), which is induced specifically under p53-dependent apoptotic conditions in human colon cancer cells, and encodes a novel polypeptide of 373 amino acids with a predicted molecular mass of 40.5 kDa. PRG3 has significant homology to bacterial oxidoreductases and the apoptosis-inducing factor, AIF, and the gene was assigned to chromosome 10q21.3-q22.1. Expression of PRG3 was induced by the activation of endogenous p53 and it contains a p53-responsive element. Unlike AIF, PRG3 localizes in the cytoplasm and its ectopic expression induces apoptosis. An amino-terminal deletion mutant of PRG3 that lacks a putative oxidoreductase activity retains its apoptotic activity, suggesting that the oxidoreductase activity is dispensable for the apoptotic function of PRG3. The PRG3 gene is thus a novel p53 target gene in a p53-dependent apoptosis pathway. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 Washington Univ, Sch Med, Dept Radiat Oncol, St Louis, MO 63108 USA. GPC Biotech Inc, Waltham, MA 02451 USA. NCI, Radiat Oncol Sci Program, Radiat Oncol Branch, NIH, Bethesda, MD 20892 USA. Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. Harvard Univ, Sch Med, Massachusetts Gen Hosp, Dept Radiat Oncol, Boston, MA 02129 USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med, Boston, MA 02215 USA. RP Horikoshi, N (reprint author), Washington Univ, Sch Med, Dept Radiat Oncol, St Louis, MO 63108 USA. RI Ohiro, Yoichi/A-5470-2012 FU NCI NIH HHS [CA75556]; NHLBI NIH HHS [R01 HL062458, HL62458] NR 57 TC 51 Z9 57 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD JUL 31 PY 2002 VL 524 IS 1-3 BP 163 EP 171 AR PII S0014-5793(02)03049-1 DI 10.1016/S0014-5793(02)03049-1 PG 9 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 580DM UT WOS:000177219500031 PM 12135761 ER PT J AU Kim, CH Park, YS Chung, KN Elwood, PC AF Kim, CH Park, YS Chung, KN Elwood, PC TI Sorting and function of the human folate receptor is independent of the caveolin expression in Fisher rat thyroid epithelial cells SO JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY LA English DT Article DE caveolin; folate receptor; Fischer rat thyroid epithelial (FRT) cell; GPI tailed protein ID PROTEIN-KINASE-C; ANCHORED PROTEINS; BINDING PROTEINS; PHOSPHOLIPASE D1; KB CELLS; ALPHA; SURFACE; GLUT4; INTERNALIZATION; MICRODOMAINS AB Caveolae are small, flask-shaped, non-clathrin coated invaginations of the plasma membrane of many mammalian cells. Caveolae have a coat that includes caveolin. They have been implicated in numerous cellular processes, including potocytosis. Since the human folate receptor (hFR) and other glycosyl-phosphatidylinositol (GPI)-tailed proteins have been co-localized to caveolae, we studied the caveolin role in the hFR function by transfecting hFR and/or caveolin cDNA into Fischer rat thyroid epithelial (FRT) cells that normally do not express detectable levels of either protein. We isolated and characterized stable clones as follows: they express (1) high levels of caveolin alone, (2) hFR and caveolin, or (3) hFR alone. We discovered that hFR is correctly processed, sorted, and anchored by a GPI tail to the plasma, membrane in FRT cells. No difference in the total folic acid binding or cell surface folic acid binding activity were found between the FRT cells that were transfected with hFR, or cells, that, were transfected with hFR and caveolin. The hFR, that was expressed on the cell surface of clones that were transfected with hFR was also sensitive to phosphaddylinositol-specific phospholipase C (PI-PLC) release, and incorporated radiolabeled ethanolamine that supports the attachment of a GPI-tail on hFR. We conclude that the processing, sorting, and function of hFR is independent on the caveolin expression in FRT cells. C1 Wonkwang Hlth Sci Coll, Dept Clin Pathol, Iksan 570750, South Korea. Wonkwang Univ, Div Biol Sci, Iksan 570749, South Korea. NIH, Bethesda, MD 20892 USA. RP Kim, CH (reprint author), Wonkwang Hlth Sci Coll, Dept Clin Pathol, Iksan 570750, South Korea. NR 37 TC 13 Z9 14 U1 0 U2 0 PU SPRINGER-VERLAG SINGAPORE PTE LTD PI SINGAPORE PA #04-01 CENCON I, 1 TANNERY RD, SINGAPORE 347719, SINGAPORE SN 1225-8687 J9 J BIOCHEM MOL BIOL JI J. Biochem. Mol. Biol. PD JUL 31 PY 2002 VL 35 IS 4 BP 395 EP 402 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 579JB UT WOS:000177173900008 PM 12296999 ER PT J AU Gonzalez, FJ AF Gonzalez, FJ TI The peroxisome proliferator-activated receptor alpha (PPAR alpha): role in hepatocarcinogenesis SO MOLECULAR AND CELLULAR ENDOCRINOLOGY LA English DT Article; Proceedings Paper CT Estrogen 2001 Congress CY MAY 21-22, 2001 CL MODENA, ITALY DE peroxisome proliferators; PPAR alpha; apoptosis; oxidative stress; hepatocarcinogenesis; Kupffer cells; cytokines ID KUPFFER CELLS; MOUSE-LIVER; FUNCTIONAL-CHARACTERIZATION; HEPATOCYTE PROLIFERATION; TARGETED DISRUPTION; SPECIES-DIFFERENCES; GENE-EXPRESSION; RAT HEPATOCYTES; DNA-SYNTHESIS; IN-VIVO AB The peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the nuclear receptor superfamily and mediates most of the known biological effects of peroxisome proliferators. The latter represents a large group of chemicals that include the fibrate hyperlipidemic drugs, the pthalate plasticizers, various solvents and degreasing agents, and endogenous hormones and fatty acids. Peroxisome proliferators are classical members of the nongenotoxic group of chemical carcinogens that do not require metabolic activation to electrophiles in order to exert their harmful effects. These chemicals are of particular concern to regulatory agencies since they can only be detected by long-term carcinogen bioassays using rodents. The mechanism of the carcinogenic action of peroxisome proliferators is beginning to emerge. PPARalpha-null mice are resistant to hepatocarcinogenesis indicating that this receptor is necessary for cancer. However, recent studies indicate that Kupffer cells, in a PPARalpha independent manor, are required for the major effects of peroxisome proliferators on cell proliferation. An interaction between PPARalpha and estrogen carcinogenesis has also been elucidated. (C) 2002 Published by Elsevier Science Ireland Ltd. C1 NCI, NIH, Bethesda, MD 20892 USA. RP Gonzalez, FJ (reprint author), NCI, NIH, Bldg 37,Room 3E-24, Bethesda, MD 20892 USA. NR 73 TC 47 Z9 48 U1 0 U2 4 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0303-7207 J9 MOL CELL ENDOCRINOL JI Mol. Cell. Endocrinol. PD JUL 31 PY 2002 VL 193 IS 1-2 SI SI BP 71 EP 79 AR PII S0303-7207(02)00098-9 DI 10.1016/S0303-7207(02)00098-9 PG 9 WC Cell Biology; Endocrinology & Metabolism SC Cell Biology; Endocrinology & Metabolism GA 584UV UT WOS:000177487900011 PM 12161004 ER PT J AU Elias, AF Schmutzhard, J Steward, PE Schwan, TG Rosa, P AF Elias, AF Schmutzhard, J Steward, PE Schwan, TG Rosa, P TI Population dynamics of a heterogenous Borrelia burgdorferi B31 strain in an experimental mouse-tick infectious cycle SO WIENER KLINISCHE WOCHENSCHRIFT LA English DT Article; Proceedings Paper CT Symposium on Tick-Transmitted Diseases CY SEP, 2001 CL LJUBLJANA, SLOVENIA DE Borrelia burgdorferi; infectious cycle; ticks; OspB; plasmids ID LYME-DISEASE SPIROCHETE; OUTER SURFACE PROTEIN; INVITRO CULTIVATION; PLASMID; ANTIBODY; MARKER; AGENT; OSPB; MICE AB We have recently shown that low-passage, infectious Borrelia burgdorferi strain B31 M I is a heterogenous mixture of clones varying in colony morphology, growth rate, protein profiles, plasmid content and infectivity. In this study, we asked whether there is a selection for certain clonal populations during the infectious cycle when uncloned B31 MI is used as the starting strain. B31 MI derivatives were reisolated from various tissues of two mice after completion of a mouse-tick-mouse infectious cycle and their protein and plasmid profiles were analyzed. Both mice developed ostensibly clonal infections despite the fact that the infectious cycle was started with a heterogenous strain. Moreover, the mice became infected with two different clones varying in protein profile and growth phenotype. Comparison of the mouse reisolates to uncloned B31 MI and clonal variants derived from B31 MI before mouse-tick-mouse passage suggests that they were derived from clonal populations present in the uncloned B31 MI. Our results indicate the presence of at least two distinct populations within B31 Mill that are competent to complete an experimental mouse-tick infectious cycle. The study provides insight into infectivity profiles and infection dynamics of different clonal populations present in a low-passage, infectious B. burgdorferi strain. C1 NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. RP Elias, AF (reprint author), Univ Klinikum Berlin, Charite, Inst Mikrobiol & Hyg, Campus Virchow Klinikum,Augustenburger Pl 1, D-13353 Berlin, Germany. NR 20 TC 7 Z9 7 U1 2 U2 2 PU SPRINGER-VERLAG WIEN PI VIENNA PA SACHSENPLATZ 4-6, PO BOX 89, A-1201 VIENNA, AUSTRIA SN 0043-5325 J9 WIEN KLIN WOCHENSCHR JI Wien. Klin. Wochen. PD JUL 31 PY 2002 VL 114 IS 13-14 BP 557 EP 561 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 581HV UT WOS:000177286700018 PM 12422601 ER PT J AU Dangi, B Dobrodumov, AV Louis, JM Gronenborn, AM AF Dangi, B Dobrodumov, AV Louis, JM Gronenborn, AM TI Solution structure and dynamics of the human - Escherichia coli thioredoxin chimera: Insights into thermodynamic stability SO BIOCHEMISTRY LA English DT Article ID HETERONUCLEAR NMR-SPECTROSCOPY; SIDE-CHAIN; BACKBONE DYNAMICS; LARGER PROTEINS; STAPHYLOCOCCAL NUCLEASE; RELAXATION MEASUREMENTS; CHEMICAL-SHIFT; J-COUPLINGS; RESOLUTION; N-15 AB We have determined the high-resolution solution structure of the oxidized form of a chimeric human and Escherichia coli thioredoxin (TRXHE) by NMR. The overall structure is well-defined with a rms difference for the backbone atoms of 0.27 +/- 0.06 Angstrom. The topology of the protein is identical to those of the human and E. coli parent proteins, consisting of a central five-stranded beta-sheet surrounded by four alpha-helices. Analysis of the interfaces between the two domains derived from the human and E. coli sequences reveals that the general hydrophobic packing is unaltered and only subtle changes in the details of side chain interactions are observed. The packing of helix alpha(4) with helix alpha(2) across the hybrid interface is less optimal than in the parent molecules, and electrostatic interactions between polar side chains are missing. In particular, lysine-glutamate salt bridges between residues on helices alpha(2) and alpha(4), which were observed in both human and E. coli proteins, are not present in the chimeric protein. The origin of the known reduced thermodynamic stability of TRXHE was probed by mutagenesis on the basis of these structural findings. Two mutants of TRXHE, S44D and S44E, were created, and their thermal and chemical stabilities were examined. Improved stability toward chaotropic agents was observed for both mutants, but no increase in the denaturation temperature was seen compared to that of TRXHE, In addition to the structural analysis, the backbone dynamics of TRXHE were investigated by N-15 NMR relaxation measurements. Analysis using the model free approach reveals that the protein is fairly rigid with an average S-2 of 0.88. Increased mobility is primarily present in two external loop regions comprising residues 72-74 and 92-94 that contain glycine and proline residues. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Gronenborn, AM (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5,Room 130, Bethesda, MD 20892 USA. OI Gronenborn, Angela M/0000-0001-9072-3525 NR 55 TC 11 Z9 12 U1 0 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 30 PY 2002 VL 41 IS 30 BP 9376 EP 9388 DI 10.1021/bi0258501 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 578HP UT WOS:000177111600013 PM 12135359 ER PT J AU Kim, S Bondeva, T Nelson, PG AF Kim, S Bondeva, T Nelson, PG TI Activation of protein kinase C isozymes in primary mouse myotubes by carbachol SO DEVELOPMENTAL BRAIN RESEARCH LA English DT Article DE PKC-alpha; PKC-theta; nicotinic acetylcholine receptor; myotube; carbachol ID ACETYLCHOLINE-RECEPTOR CLUSTERS; CULTURED RAT MYOTUBES; NEUROMUSCULAR-JUNCTION; SIGNAL-TRANSDUCTION; PHORBOL ESTER; MUSCLE-CELLS; SKELETAL-MUSCLE; ALPHA-ACTININ; DELTA-SUBUNIT; PHOSPHORYLATION AB The activation of muscle PKC isozymes following treatment with carbachol, an acetylcholine receptor agonist, has been investigated. Primary mouse myotubes were treated with carbachol, and protein extracts from the cytosol and membrane fractions of the myotubes were subjected to Western blot analyses. Carbachol treatment resulted in a rapid translocation of PKC-theta to the membrane. This effect was dependent on both carbachol concentration and incubation time. The treatment also resulted in a drastic increase of PKC-alpha in the cytosol followed by an increase of PKC-alpha in the membrane. The regulation of PKC-alpha in response to carbachol was quite distinct from that produced by the PKC activator, PMA, which rapidly translocated PKC-alpha from the cytosol to the membrane without any increases in PKC-alpha in the cytosol. Confocal microscopy demonstrated an enhanced membrane localization of PKC-theta and overall increased intensity of PKC-alpha staining in the cyrosol accompanied by a characteristic membrane staining of PKC-alpha in the myotubes treated with carbachol. Taken together, the results suggested that the activation of PKC isozymes in response to the receptor agonist is quite distinct, which indicates their diverse role in the muscle upon the release of neurotransmitter at the neuromuscular junction. C1 NICHD, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Kim, S (reprint author), Human Genome Sci Inc, 9410 Key W Ave, Rockville, MD 20850 USA. NR 38 TC 11 Z9 11 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-3806 J9 DEV BRAIN RES JI Dev. Brain Res. PD JUL 30 PY 2002 VL 137 IS 1 BP 13 EP 21 AR PII S0165-3806(02)00362-0 DI 10.1016/S0165-3806(02)00362-0 PG 9 WC Developmental Biology; Neurosciences SC Developmental Biology; Neurosciences & Neurology GA 583QB UT WOS:000177418000002 ER PT J AU Donovan, DM Becker, KG AF Donovan, DM Becker, KG TI Double round hybridization of membrane based cDNA arrays: improved background reduction and data replication SO JOURNAL OF NEUROSCIENCE METHODS LA English DT Article DE cDNA; microarray; neuronal; hybridizations; radioactive; probe; background; gene expression ID MICROARRAY; EXPRESSION; CANCER AB Microarray analysis provides the ability to obtain gene expression data on thousands of gene targets in parallel and is being applied to an increasing variety of neurobiological samples. Nylon filter based cDNA arrays often provide an alternative to oligobased or glass-based gene expression analysis systems. cDNA arrays regardless of the platform, are usually quite expensive to purchase and equally expensive to produce. As such, obtaining both high quality and replicated data, as well as the ability to recover from an apparently failed hybridization experiment are important factors in microarray experimental design. Using a collection of 1152 neuronally relevant cDNAs, this report describes simple modifications in experimental design of radioactive microarray experiments that, (1) improves reproducibility; (2) allows recovery from a failed experiment and; (3) provides a type of replicate data set with no requirement for additional arrays, no additional probe labelings, and little extra cost. (C) 2002 Published by Elsevier Science B.V. C1 NIA, DNA Array Unit, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. RP Becker, KG (reprint author), NIA, DNA Array Unit, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Becker, Kevin/0000-0002-6794-6656 NR 8 TC 10 Z9 10 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-0270 J9 J NEUROSCI METH JI J. Neurosci. Methods PD JUL 30 PY 2002 VL 118 IS 1 BP 59 EP 62 AR PII S0165-0270(02)00141-3 DI 10.1016/S0165-0270(02)00141-3 PG 4 WC Biochemical Research Methods; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 590VC UT WOS:000177843800007 PM 12191758 ER PT J AU Ling, BH Veazey, RS Luckay, A Penedo, C Xu, KY Lifson, JD Marx, PA AF Ling, BH Veazey, RS Luckay, A Penedo, C Xu, KY Lifson, JD Marx, PA TI SIVmac pathogenesis in rhesus macaques of Chinese and Indian origin compared with primary HIV infections in humans SO AIDS LA English DT Article DE rhesus; macaque; subspecies; SIV; HIV; viral load; intestinal effector cells ID SIMIAN IMMUNODEFICIENCY VIRUS; T-CELL DEPLETION; DISEASE PROGRESSION; LYMPHOCYTES; VACCINE; SEROCONVERSION; REPLICATION; RETROVIRUS; EXPRESSION; RESPONSES AB Objective: To develop a SIV-rhesus macaque (Rh) model of AIDS that more closely approximates HIV pathogenesis in humans. Design: The pathogenesis of SIV was compared in two different types of Rh, the Chinese (Ch) and Indian (Ind) subspecies. Methods: Ch Rh and Ind Rh origin were identified genetically and infected with the SIVmac239 molecular clone. Plasma viral loads, depletion of intestinal lymphocytes with memory phenotype, humoral immune responses and CD4/CD8 cell ratios were compared during acute and steady-state periods of infection. Results: Plasma viral loads from 7 days after infection through 240 days were significantly lower in Rh of Ch origin compared with Ind Rh. Viral loads in Ch Rh were closer to viral loads observed in untreated humans infected with HIV-1. Depletion of intestinal effector cells was less evident in SIV-infected Ch Rh compared with Ind Rh. An index of intestinal pathogenesis was devised that closely paralleled viral load and severity of infection, There were no rapid progressors to AIDS among 10 Ch Rh. In contrast, three of four Ind Rh progressed rapidly to AIDS. Conclusions: Compared with Ind Rh, SIVmac pathogenesis in Ch Rh was closer to HIV-1 infections in untreated adult humans. The differences were statistically significant. The Ch Rh subspecies is a suitable AIDS model and may have advantages over the rapid and highly pathogenic Ind Rh model. Moreover, Ind Rh supplies are limited and use of Ch Rh provides a new resource. (C) 2002 Lippincott Williams Wilkins. C1 Tulane Univ, Hlth Sci Ctr, Dept Trop Med, Covington, LA USA. Tulane Reg Primate Res Ctr, Covington, LA USA. Rockefeller Univ, Aaron Diamond AIDS Res Ctr, New York, NY 10021 USA. Univ Calif Davis, Vet Genet Lab, Davis, CA 95616 USA. NCI, FCRDC, AIDS Vaccine Program, Frederick, MD 21701 USA. RP Marx, PA (reprint author), Tulane Nalt Primate Res Ctr, 18703 3 Rivers Rd, Covington, LA 70433 USA. FU NCRR NIH HHS [P51 RR000164, P51RR000164-40]; NIAID NIH HHS [R01 AI45510]; NICHD NIH HHS [HD36310] NR 26 TC 150 Z9 160 U1 0 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 26 PY 2002 VL 16 IS 11 BP 1489 EP 1496 DI 10.1097/00002030-200207260-00005 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 578HT UT WOS:000177111900005 PM 12131186 ER PT J AU Chowdhury, B Mantile-Selvaggi, G Miele, L Cordella-Miele, E Zhang, ZJ Mukherjee, AB AF Chowdhury, B Mantile-Selvaggi, G Miele, L Cordella-Miele, E Zhang, ZJ Mukherjee, AB TI Lys 43 and Asp 46 in alpha-helix 3 of uteroglobin are essential for its phospholipase A(2)-inhibitory activity SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID RECOMBINANT HUMAN UTEROGLOBIN; CELL 10-KDA PROTEIN; GENE-EXPRESSION; ANTIFLAMMIN PEPTIDES; RABBIT UTEROGLOBIN; BINDING PROTEINS; LIGAND-BINDING; PURIFICATION; INHIBITION; CHEMOTAXIS AB Uteroglobin (UG) is an anti-inflammatory, secreted protein with soluble phospholipase A, (sPLA,)-inhibitory activity. However, the mechanism by which UG inhibits sPLA, activity is unknown. UG is a homodimer in which each of the 70-amino acid subunits forms four alpha-helices. We previously reported that sPLA(2)-inhibitory activity of UG may reside in a segment of alpha-helix 3 that is exposed to the solvent. In addition. it has been suggested that UG may inhibit sPLA(2) activity by binding and sequestering Ca-`(++) essential for sPLA(2) activation. By site-specific mutation, we demonstrate here that Lys 43 Glu Asp 46 Lys or a combination of the two mutations in the full-length, recombinant human UG (rhUG) abrogates its sPLA(2)-inhibitory activity. We demonstrate further that recombinant UG does not bind Ca++ although when it is expressed with histidine-tag (H-tag) it is capable of binding Ca+-. Taken together our results show that: (i) Lys 43 and Asp 46 in rhUG are critical residues for the sPLA(2)-inhibitory activity of UG and (ii) Ca++-sequestration by rhUG is not likely to be one of the mechanisms responsible for its sPLA(2)-inhibitory activity. (C) 2002 Elsevier Science (USA). All rights reserved. C1 NICHD, Sect Dev Genet, Heritable Disorders Branch, Bethesda, MD 20892 USA. RP Mukherjee, AB (reprint author), NICHD, Sect Dev Genet, Heritable Disorders Branch, Bethesda, MD 20892 USA. NR 51 TC 13 Z9 13 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUL 26 PY 2002 VL 295 IS 4 BP 877 EP 883 AR PII S0006-291X(02)00767-2 DI 10.1016/S0006-291X(02)00767-2 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 580XL UT WOS:000177262000019 PM 12127976 ER PT J AU Choung, PH Seo, BM Chung, CP Yamada, KM Jang, JH AF Choung, PH Seo, BM Chung, CP Yamada, KM Jang, JH TI Synergistic activity of fibronectin and fibroblast growth factor receptors on neuronal adhesion and neurite extension through extracellular signal-regulated kinase pathway SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE FGFR; FGF; fibronectin; integrin; neurite outgrowth ID MAP KINASE; TYROSINE KINASES; ACTIVATION; CELLS; SPECIFICITY; INTEGRINS; TRANSIENT; SEQUENCE; CULTURE; FAMILY AB Fibroblast growth factor (FGF) is an important modulator of cell growth and differentiation of various Cells including neuron. Cells need to adhere specifically to cellular and extracellular components of their environment to carry out diverse physiological functions. Here. we examined whether fibronectin (FN) and FGF can cooperate for neuronal adhesion and neurite outgrowth. Using recombinant FN peptide (FNIII9-10), we found that FNIII9-10-mediated adhesion promotes the effect of FGF1 on neurite outgrowth of PC12 cells, while FGF1 enhances the FNIII9-10-mediated neuronal adhesion of PC12 cells. This collaboration of FNIII9-10 and FGF1 was the result of the sustained activation of extracellular signal-regulated kinase (ERK)-type MAP kinase, Finally. the synergistic activity of FGF1 and FN was inhibited by PD98059, an MEK inhibitor. Taken together, these findings indicate that FN-mediated signaling can collaborate with FGFRs signaling for neurite outgrowth through selective activation of ERK-type MAP kinase in PC12 cells. and suggest that FN and FGF act in concert to regulate cell differentiation in the nervous system. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Seoul Natl Univ, Coll Dent, Intellectual Biointerface Engn Ctr, Seoul 110768, South Korea. Seoul Natl Univ, Coll Dent, Dept Oral & Maxillofacial Surg, Craniofacial Tissue Engn Lab BK21 Human Life Sci, Seoul 110768, South Korea. Seoul Natl Univ, Coll Dent, Dept Periodontol, Seoul 110768, South Korea. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. RP Jang, JH (reprint author), Seoul Natl Univ, Coll Dent, Intellectual Biointerface Engn Ctr, 28 Yongon Dong, Seoul 110768, South Korea. OI Yamada, Kenneth/0000-0003-1512-6805 NR 26 TC 18 Z9 19 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUL 26 PY 2002 VL 295 IS 4 BP 898 EP 902 AR PII S0006-291X(02)00774-X DI 10.1016/S0006-291X(02)00774-X PG 5 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 580XL UT WOS:000177262000022 PM 12127979 ER PT J AU Gustchina, A Li, M Phylip, LH Lees, WE Kay, J Wlodawer, A AF Gustchina, A Li, M Phylip, LH Lees, WE Kay, J Wlodawer, A TI An unusual orientation for Tyr75 in the active site of the aspartic proteinase from Saccharomyces cerevisiae SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE aspartic proteinases; active site; protein structure; structure comparison; crystal packing ID COMPLEMENT FACTOR-D; CRYSTAL-STRUCTURE; 3-DIMENSIONAL STRUCTURE; RETROVIRAL PROTEASES; ANGSTROM RESOLUTION; BOVINE CHYMOSIN; SERINE-PROTEASE; SPECIFICITY; INHIBITOR; ZYMOGEN AB The structures of the native Saccharomyces cerevisiae proteinase A have been solved by molecular replacement in the monoclinic and trigonal crystal forms and refined at 2.6-2.7 A resolution. These structures agree overall with those of other uninhibited aspartic proteinases. However, an unusual orientation for the side chain of Tyr75, a conserved residue on the flexible "flap" that covers the active site and is important for the activity of these enzymes. was found in the trigonal crystals. A similar conformation of Tyr75 occupying the S-1 substrate-binding pocket was previously reported only for chymosin (where it was interpreted as representing a "self-inhibited" state of the enzyme). but for no other aspartic proteinases. Since this orientation of Tyr75 has now been seen in the structures of two members of the family of aspartic proteinases, it might indicate that the placement of that residue in the S, substrate-binding pocket might have some functional significance. analogous to what was seen for self-inhibited structures of serine proteinases. (C) 2002 Elsevier Science (USA). All rights reserved. C1 NCI, Prot Struct Sect, Macromol Crystallog Lab, Frederick, MD 21702 USA. NCI, Intramural Res Support Program, Sci Applicat Int Corp, Frederick, MD 21702 USA. Cardiff Univ, Sch Biosci, Cardiff CF10 3US, S Glam, Wales. RP Wlodawer, A (reprint author), NCI, Prot Struct Sect, Macromol Crystallog Lab, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 29 TC 14 Z9 14 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUL 26 PY 2002 VL 295 IS 4 BP 1020 EP 1026 AR PII S0006-291X(02)00742-8 DI 10.1016/S0006-291X(02)00742-8 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 580XL UT WOS:000177262000041 PM 12127998 ER PT J AU Renstrom, E Ivarsson, R Shears, SB AF Renstrom, E Ivarsson, R Shears, SB TI Inositol 3,4,5,6-tetrakisphosphate inhibits insulin granule acidification and fusogenic potential SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CELL-LINE; CHLORIDE CHANNELS; CYSTIC-FIBROSIS; CALCIUM; SECRETION; INS(3,4,5,6)P-4; CONDUCTANCE; EXOCYTOSIS; GLUCOSE AB CIC Cl- channels in endosomes, synaptosomes, lysosomes, and beta-cell insulin granules provide charge neutralization support for the functionally indispensable acidification of the luminal interior by electrogenic H+-ATPases (Jentsch, T. J., Stein, V., Weinreich, F., and Zdebik, A. A. (2002) Physiol Rev. 82,503-568). Regulation of CIC activity is, therefore, of widespread biological significance (Forgac, M. (1999) J. Biol Chem. 274,1295112954). We now ascribe just such a regulatory function to the increases in cellular levels of inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P-4) that inevitably accompany activation of the ubiquitous Ins(1,4,5)P-3 signaling pathway. We used confocal. imaging to record insulin granule acidification in single mouse pancreatic beta-cells. Granule acidification was reduced by perfusion of single cells with 10 muM lns(3,4,5,6)P-4 (the concentration following receptor activation), whereas at 1 pm ("resting" levels), Ins(3,4,5,6)P-4 was ineffective. This response to lns(3,4,5,6)P-4 was not mimicked by 100 muM lns(1,4,5,6)P4 or by 100 muM Ins(1,3,4,5,6)P-5. Ins(3,4,5,6)P-4 did not affect granular H+-ATPase activity or H+ leak, indicating that Ins(3,4,5,6)P4 instead inhibited charge neutralization by CIC. The lns(3,4,5,6)P-4-mediated inhibition of vesicle acidification reduced exocytic release of insulin as determined by whole-cell capacitance recordings. This may impinge upon type 2 diabetes etiology. Regulatory control over vesicle acidification by this negative signaling pathway in other cell types should be considered. C1 Lund Univ, Dept Physiol Sci, SE-22184 Lund, Sweden. NIEHS, Inositide Signaling Grp, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Renstrom, E (reprint author), Lund Univ, Dept Physiol Sci, BMC F11, SE-22184 Lund, Sweden. NR 27 TC 22 Z9 23 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 26717 EP 26720 DI 10.1074/jbc.C200314200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900002 PM 12055181 ER PT J AU Zakalskiy, A Hogenauer, G Ishikawa, T Wehrschutz-Sigl, E Wendler, F Teis, D Zisser, G Steven, AC Bergler, H AF Zakalskiy, A Hogenauer, G Ishikawa, T Wehrschutz-Sigl, E Wendler, F Teis, D Zisser, G Steven, AC Bergler, H TI Structural and enzymatic properties of the AAA protein Drg1p from Saccharomyces cerevisiae - Decoupling of intracellular function from ATPase activity and hexamerization SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SENSITIVE FUSION PROTEIN; ESCHERICHIA-COLI; MEMBRANE-FUSION; CLPAP PROTEASE; BINDING SITES; FAMILY; YEAST; TRANSPORT; HOMOLOGY; DISTINCT AB The AAA protein Drg1 from yeast was affinity-purified, and its ATPase activity and hexamerization properties were analyzed. The same parameters were also determined for several mutant proteins and compared in light of the growth characteristics of the corresponding cells. The protein from a thermosensitive mutant exhibited reduced ATPase activity and hexamerization. These defects were not reversed by an intragenic suppressor mutation, although this allele supported growth at the nonpermissive temperature. A different set of mutants was generated by site-specific mutagenesis intended to adjust the Walker A box of the D2 domain of Drg1p to that of the D1 domain. A S562G exchange in D2 produced a nonfunctional protein that did not hexamerize but showed above-normal ATPase activity. The C561T mutant protein, on the other hand, was functional but hexamerized less readily and had reduced ATPase activity. In contrast, the C561T/S562G protein hexamerized less than wild type but had much higher ATPase activity. We distinguished strong and weak ATP-binding sites in the wild type protein but two weak sites in the C561T/S562G protein, indicating that the stronger site resides in D2. These observations are discussed in terms of the inter-relationship of ATPase activity per se, oligomeric status, and intracellular function for AAA proteins. C1 Karl Franzens Univ Graz, Inst Mol Biol Biochem & Mikrobiol, A-8010 Graz, Austria. NIAMS, Struct Biol Lab, NIH, Bethesda, MD 20892 USA. RP Hogenauer, G (reprint author), Karl Franzens Univ Graz, Inst Mol Biol Biochem & Mikrobiol, Univ Pl 2, A-8010 Graz, Austria. RI Ishikawa, Takashi/E-5023-2017; OI Ishikawa, Takashi/0000-0002-1976-7477; Bergler, Helmut/0000-0002-7724-309X NR 30 TC 19 Z9 19 U1 1 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 26788 EP 26795 DI 10.1074/jbc.M201515200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900012 PM 12006565 ER PT J AU Fehlner-Gardiner, C Roshick, C Carlson, JH Hughes, S Belland, RJ Caldwell, HD McClarty, G AF Fehlner-Gardiner, C Roshick, C Carlson, JH Hughes, S Belland, RJ Caldwell, HD McClarty, G TI Molecular basis defining human Chlamydia trachomatis tissue tropism - A possible role for tryptophan synthase SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID COMPLETE GENOME SEQUENCE; OUTER-MEMBRANE PROTEIN; ESCHERICHIA-COLI; ALPHA-SUBUNIT; SALMONELLA-TYPHIMURIUM; GAMMA-INTERFERON; FLEXIBLE LOOP; ALPHA-2-BETA-2 COMPLEX; NUCLEOTIDE-SEQUENCE; BETA-2 SUBUNIT AB Here we report the cloning and sequencing of a region of the chlamydiae chromosome termed the "plasticity zone" from all the human serovars of C. trachomatis containing the tryptophan biosynthesis genes. Our results show that this region contains orthologues of the tryptophan repressor as well as the alpha and beta subunits of tryptophan synthase. Results from reverse transcription-PCR and Western blot analyses indicate that the trpBA genes are transcribed, and protein products are expressed. The TrpB sequences from all serovars are highly conserved. In comparison with other tryptophan synthase beta subunits, the chlamydial TrpB subunit retains all conserved amino acid residues required for beta reaction activity. In contrast, the chlamydial TrpA sequences display numerous mutations, which distinguish them from TrpA sequences of all other prokaryotes. All ocular serovars contain a deletion mutation resulting in a truncated TrpA protein, which lacks a reaction activity. The TrpA protein from the genital serovars retains conserved amino acids required for catalysis but has mutated several active site residues involved in substrate binding. Complementation analysis in Escherchia coli strains, with defined mutations in tryptophan biosynthesis, and in vitro enzyme activity data, with cloned TrpB and TrpA proteins, indicate these mutations result in a TrpA protein that is unable to utilize indole glycerol 3-phosphate as substrate. In contrast, the chlamydial TrpB protein can carry out the betareaction, which catalyzes the formation of tryptophan from indole and serine. The activity of the chlamydial Trp B protein differs from that of the well characterized E. coli and Salmonella TrpBs in displaying an absolute requirement for full-length TrpA. Taken together our data indicate that genital, but not ocular, serovars are capable of utilizing exogenous indole for the biosynthesis of tryptophan. C1 Univ Manitoba, Dept Med Microbiol, Winnipeg, MB R3E 0W3, Canada. Hlth Canada, Natl Microbiol Lab, Winnipeg, MB R3E 0W3, Canada. NIAID, Rocky Mt Labs, Intracellular Parasites Lab, NIH, Hamilton, MT 59840 USA. NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. RP McClarty, G (reprint author), Univ Manitoba, Dept Med Microbiol, Winnipeg, MB R3E 0W3, Canada. EM mcclart@cc.umanitoba.ca NR 67 TC 97 Z9 97 U1 0 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 26893 EP 26903 DI 10.1074/jbc.M203937200 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900027 PM 12011099 ER PT J AU Sweet, DH Miller, DS Pritchard, JB Fujiwara, Y Beier, DR Nigam, SK AF Sweet, DH Miller, DS Pritchard, JB Fujiwara, Y Beier, DR Nigam, SK TI Impaired organic anion transport in kidney and choroid plexus of organic anion transporter 3 (Oat3 (Slc22a8)) knockout mice SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID OSTEOSCLEROSIS OC MUTATION; MOLECULAR-CLONING; RAT-LIVER; FUNCTIONAL-CHARACTERIZATION; EXPRESSION CLONING; EPITHELIAL-CELLS; LOCALIZATION; BRAIN; IDENTIFICATION; PROTEIN AB To begin to develop in vivo model systems for the assessment of the contributions of specific organic anion transporter (OAT) family members to detoxification, development, and disease, we carried out a targeted disruption of the murine organic anion transporter 3 (Oat3) gene. Surviving Oat3(-/-) animals appear healthy, are fertile, and do not exhibit any gross morphological tissue abnormalities. No Oat3 mRNA expression was detected in kidney, liver, or choroid plexus (CP) of Oat3(-/-) mice. A distinct phenotype manifested by a substantial loss of organic anion transport capacity in kidney and CP was identified. Uptake sensitive to inhibition by bromosulfophthalein or probenecid was observed for taurocholate, estrone sulfate, and para-aminohippurate in renal slices from wild-type mice, whereas in Oat3(-/-) animals transport of these substances was greatly reduced. No discernable differences in uptake were observed between hepatic slices from wild-type and Oat(-/-) littermates, suggesting Oat3 does not play a major role in hepatic organic anion uptake. Cellular accumulation of fluorescein was reduced by similar to75% in CP from Oat3(-/-) mice. However, capillary accumulation of fluorescein-methotrexate was unchanged, indicating the effects of Oat3 loss are restricted to the entry step and that Oat3 is localized to the apical membrane of CP. These data indicate a key role for Oat3 in systemic detoxification and in control of the organic anion distribution in cerebrospinal fluid. C1 Univ Calif San Diego, Dept Pediat, Div Nephrol Hypertens, La Jolla, CA 92093 USA. Univ Calif San Diego, Dept Med, Div Nephrol Hypertens, La Jolla, CA 92093 USA. Univ Calif San Diego, Dept Cellular & Mol Med, La Jolla, CA 92093 USA. NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. Harvard Univ, Childrens Hosp, Howard Hughes Med Inst, Div Hematol Oncol,Sch Med, Boston, MA 02115 USA. Harvard Univ, Brigham & Womens Hosp, Sch Med, Div Genet, Boston, MA 02115 USA. RP Nigam, SK (reprint author), Univ Calif San Diego, Dept Pediat, Div Nephrol Hypertens, 9500 Gilman Dr 0693, La Jolla, CA 92093 USA. RI Sweet, Douglas/H-7914-2013 OI Sweet, Douglas/0000-0002-8911-9184 FU NICHD NIH HHS [R01 HD 40011] NR 47 TC 194 Z9 201 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 26934 EP 26943 DI 10.1074/jbc.M203803200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900032 PM 12011098 ER PT J AU Erxleben, C Everhart, AL Romeo, C Florance, H Bauer, MB Alcorta, DA Rossie, S Shipston, MJ Armstrong, DL AF Erxleben, C Everhart, AL Romeo, C Florance, H Bauer, MB Alcorta, DA Rossie, S Shipston, MJ Armstrong, DL TI Interacting effects of N-terminal variation and strex exon splicing on slo potassium channel regulation by calcium, phosphorylation, and oxidation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CA2+-ACTIVATED K+ CHANNELS; LARGE-CONDUCTANCE; SMOOTH-MUSCLE; BETA-SUBUNIT; PROTEIN-PHOSPHORYLATION; XENOPUS-OOCYTES; BKCA CHANNEL; CA CHANNELS; MODULATION; CELLS AB We have investigated the structural basis for the phenotype of a native rat Slo (rSlo) potassium channel (BKCa; KCNMA1) in a rat pituitary cell line, GH(4)C(1). Opposing regulation of these calcium- and voltage-activated potassium channels by cAMP- and cGMP-dependent protein kinases requires an alternatively spliced exon (strex) of 59 amino acids in the cytoplasmic C terminus of the pore-forming alpha subunit encoded by rslo. However, inclusion of this cysteine-rich exon produces a 10-fold increase in the sensitivity of the channels to inhibition by oxidation. Inclusion of the strex exon also increases channel sensitivity to stimulation by calcium, but responses in the physiological ranges of calcium and voltage require coassembly with beta(1) subunits. With strex present, however, beta(1) subunits only stimulated channels assembled from rSlo alpha subunits with a truncated N terminus beginning MDALI-. Thus N-terminal variation and strex exon splicing in rSlo interact to produce BKCa channels with a physiologically relevant phenotype. C1 NIEHS, Lab Signal Transduct F205, NIH, Res Triangle Pk, NC 27709 USA. Univ Edinburgh, Membrane Biol Grp, Edinburgh EH8 9XD, Midlothian, Scotland. Purdue Univ, Dept Biochem, W Lafayette, IN 47907 USA. RP Armstrong, DL (reprint author), NIEHS, Lab Signal Transduct F205, NIH, 111 Alexander Dr, Res Triangle Pk, NC 27709 USA. RI Shipston, Mike/C-7309-2013 OI Shipston, Mike/0000-0001-7544-582X NR 44 TC 46 Z9 49 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27045 EP 27052 DI 10.1074/jbc.M203087200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900046 PM 12016222 ER PT J AU Chikumi, H Vazquez-Prado, J Servitja, JM Miyazaki, H Gutkind, JS AF Chikumi, H Vazquez-Prado, J Servitja, JM Miyazaki, H Gutkind, JS TI Potent activation of RhoA by G alpha(q) and G(q)-coupled receptors SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GUANINE-NUCLEOTIDE EXCHANGE; PROTEIN-COUPLED RECEPTORS; HETEROTRIMERIC G-PROTEINS; STRESS FIBER FORMATION; P115 RHOGEF; KINASE; ADHESION; GROWTH; FAMILY; RAS AB Heterotrimeric G proteins of the G(i), G(s), and G(q) family control a wide array of physiological functions primarily by regulating the activity of key intracellular second messenger-generating systems. a subunits of the G(12) family, Galpha(12) and Galpha(13), however, can promote cellular responses that are independent of conventional second messengers but that result from the activation of small GTP-binding proteins of the Rho family and their downstream targets. These findings led to the identification of a novel family of guanine-nucleotide exchange factors (GEFs) that provides a direct link between Galpha(12/13) and Rho stimulation. Recent observations suggest that many cellular responses elicited by Galpha(q) and its coupled receptors also require the functional activity of Rho. However, available evidence suggests that Gaq may act on pathways downstream from Rho rather than by promoting Rho activation. These seemingly conflicting observations and the recent development of sensitive assays to assess the in vivo levels of active Rho prompted us to ask whether Gaq and its coupled receptors can stimulate endogenous Rho. Here we show that the expression of activated forms of Gaq and the stimulation of G(q)-coupled receptors or chimeric Galpha(q) molecules that respond to G(i)-linked receptors can promote a robust activation of endogenous Rho in HEK-293T cells. Interestingly, this response was not prevented by molecules interfering with the ability of G a,, to stimulate its linked RhoGEFs, together suggesting the existence of a novel molecular mechanism by which Gaq and the large family of G(q)-coupled receptors can regulate the activity of Rho and its downstream signaling pathways. C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Gutkind, JS (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, 9000 Rockville Pike,Bldg 30,Rm 211, Bethesda, MD 20892 USA. RI Gutkind, J. Silvio/A-1053-2009; VAZQUEZ-PRADO, JOSE/C-1630-2017; OI SERVITJA DUQUE, JOAN-MARC/0000-0002-1241-8004 NR 24 TC 116 Z9 116 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27130 EP 27134 DI 10.1074/jbc.M204715200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900056 PM 12016230 ER PT J AU Gu, JY Park, JM Song, EJ Mizuguchi, G Yoon, JH Kim-Ha, J Lee, KJ Kim, YJ AF Gu, JY Park, JM Song, EJ Mizuguchi, G Yoon, JH Kim-Ha, J Lee, KJ Kim, YJ TI Novel mediator proteins of the small Mediator complex in Drosophila SL2 cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID RNA-POLYMERASE-II; SIGNAL-TRANSDUCTION PATHWAYS; TRANSCRIPTIONAL ACTIVATION; IN-VIVO; BINDING; YEAST; HOLOENZYME; GENES AB The Mediator complex is generally required for transcriptional regulation in species ranging from yeast to human. Throughout evolution, the functional diversity of the Mediator complex has been enhanced to meet the increasing requirements for sophisticated gene regulation. It is likely that greater structural complexity is thus required to accomplish these new, complex regulatory functions. In this study, we took systematic steps to examine various types of Mediator complexes in Drosophila melanogaster. Such efforts led to the identification of three distinct forms of Mediator complexes. In exploring their compositional and functional heterogeneity, we found that the smallest complex (Cl) is highly enriched in a certain type of Drosophila cells and possesses novel Mediator proteins. The subunits shared among the three Mediator complexes (C1, C2, and C3) appear to form a stable modular structure that serves as a binding surface for transcriptional activator proteins. However, only C2 and C3 were able to support activated transcription in vitro. These findings suggest that different cell types may require distinct Mediator complexes, some of which may participate in nuclear processes other than the previously identified functions. C1 Yonsei Univ, Dept Biochem, Coll Sci,Natl Creat Res Initiat Ctr Genome Regula, Seodaemoon Ku, Seoul 120749, South Korea. Digital Genom Inc, Seoul 120749, South Korea. Ewha Womans Univ, Ctr Cell Signaling Res, Div Mol Life Sci, Seoul 120750, South Korea. Ewha Womans Univ, Coll Pharm, Seoul 120750, South Korea. NCI, Mol Cell Biol Lab, NIH, Bethesda, MD 20892 USA. Sejong Univ, Dept Biol Mol, Seoul 143797, South Korea. RP Kim, YJ (reprint author), Yonsei Univ, Dept Biochem, Coll Sci,Natl Creat Res Initiat Ctr Genome Regula, Seodaemoon Ku, Sinchondong 134, Seoul 120749, South Korea. OI Lee, Kong-Joo/0000-0001-7972-6020 NR 28 TC 16 Z9 28 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27154 EP 27161 DI 10.1074/jbc.M204144200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900059 PM 12021283 ER PT J AU Inamdar, KV Pouliot, JJ Zhou, T Lees-Miller, SP Rasouli-Nia, A Povirk, LF AF Inamdar, KV Pouliot, JJ Zhou, T Lees-Miller, SP Rasouli-Nia, A Povirk, LF TI Conversion of phosphoglycolate to phosphate termini on 3 ' overhangs of DNA double strand breaks by the human tyrosyl-DNA phosphodiesterase hTdp1 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID 3'-PHOSPHOGLYCOLATE TERMINI; IONIZING-RADIATION; OXIDATIVE DAMAGE; IN-VITRO; REPAIR; NEOCARZINOSTATIN; EXONUCLEASE; MECHANISMS; BLEOMYCIN; CALICHEAMICIN AB Mammalian cells contain potent activity for removal of 3'-phosphoglycolates from single-stranded oligomers and from 3' overhangs of DNA double strand breaks, but no specific enzyme has been implicated in such removal. Fractionated human whole-cell extracts contained an activity, which in the presence of EDTA, catalyzed removal of glycolate from phosphoglycolate at a single-stranded 3' terminus to leave a 3'-phosphate, reminiscent of the human tyrosyl-DNA phosphodiesterase hTdp1. Recombinant hTdp1, as well as Saccharomyces cerevisiae Tdp1, catalyzed similar removal of glycolate, although less efficiently than removal of tyrosine. Moreover, glycolate-removing activity could be immunodepleted from the fractionated extracts by antiserum to hTdpl. When a plasmid containing a double strand break with a 3'-phosphoglycolate on a 3-base 3' overhang was incubated in human cell extracts, phosphoglycolate processing proceeded rapidly for the first few minutes but then slowed dramatically, suggesting that the single-stranded overhangs gradually became sequestered and inaccessible to hTdp1. The results suggest a role for hTdp1 in repair of free radical-mediated DNA double strand breaks bearing terminally blocked 3' overhangs. C1 Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. NIMH, Mol Biol Lab, Bethesda, MD 20892 USA. Univ Calgary, Dept Biochem & Mol Biol, Calgary, AB T2N 1N4, Canada. Univ Alberta, Dept Oncol, Edmonton, AB T6G 1Z2, Canada. RP Povirk, LF (reprint author), Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, POB 980230, Richmond, VA 23298 USA. FU NCI NIH HHS [CA 40615] NR 32 TC 112 Z9 114 U1 0 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27162 EP 27168 DI 10.1074/jbc.M204688200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900060 PM 12023295 ER PT J AU Dobson, GP Hitchins, S Teague, WE AF Dobson, GP Hitchins, S Teague, WE TI Thermodynamics of the pyruvate kinase reaction and the reversal of glycolysis in heart and skeletal muscle SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GASTROCNEMIUS IN-VIVO; CREATINE-KINASE; GLYCOGEN-SYNTHESIS; LACTATE; METABOLISM; EXERCISE; FROG; PH AB The effect of temperature, pH, and free [Mg2+] on the apparent equilibrium constant of pyruvate kinase (phosphoenol transphosphorylase) (EC 2.7.1.40) was investigated. The apparent equilibrium constant, K', for the biochemical reaction P-enolpyruvate + ADP = ATP + Pyr was defined as K' = [ATP][Pyr]/[ADP][P-enolpyruvatel] where each reactant represents the sum of all the ionic and metal complexed species in M. The K' at pH 7.0, 1.0 mm free Mg2+ and I of 0.25 m was 3.89 X 10(4) (n = 8) at 25 degreesC. The standard apparent enthalpy (DeltaH'degrees) for the biochemical reaction was -4.31 kJmol(-1) in the direction of ATP formation. The corresponding standard apparent entropy (DeltaS'degrees) was +73.4 J K-1 mol(-1). The DeltaHdegrees and AS' values for the reference reaction, P-enolpyruvate(3-) + ADp(3-) + H+ = ATp(4-) + Pyr(1-), were -6.43 kJmol(-1) and + 180 J K-1 mol(-1), respectively (5 to 38 degreesC). We examined further the mass action ratio in rat heart and skeletal muscle at rest and found that the pyruvate kinase reaction in vivo was close to equilibrium i.e. within a factor of about 3 to 6 of K' in the direction of ATP at the same pH, free [Mg2+], and T. We conclude that the pyruvate kinase reaction may be reversed under some conditions in vivo, a finding that challenges the long held dogma that the reaction is displaced far from equilibrium. C1 James Cook Univ N Queensland, Sch Biomed & Mol Sci, Div Physiol & Pharmacol, Townsville, Qld 4811, Australia. NIAAA, Sect Nucl Magnet Resonacne Studies, Lab Membrane Biochem & Biophys, Rockville, MD 20852 USA. RP Dobson, GP (reprint author), James Cook Univ N Queensland, Sch Biomed & Mol Sci, Div Physiol & Pharmacol, Townsville, Qld 4811, Australia. NR 40 TC 14 Z9 14 U1 1 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27176 EP 27182 DI 10.1074/jbc.M111422200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900062 PM 11986306 ER PT J AU Mills, EM Xu, D Fergusson, MM Combs, CA Xu, YH Finkel, T AF Mills, EM Xu, D Fergusson, MM Combs, CA Xu, YH Finkel, T TI Regulation of cellular oncosis by uncoupling protein 2 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID APOPTOSIS; DEATH; MITOCHONDRIA; OBESITY; GENE; NECROSIS; MICE; UCP2 AB Cell death can proceed through at least two distinct pathways. Apoptosis is an energy-dependent process characterized morphologically by cell shrinkage, whereas oncosis is a form of cell death induced by energy depletion and initially characterized by cell swelling. We demonstrate in HeLa cells but not in normal diploid fibroblasts that modest increases in the expression level of uncoupling protein 2 (UCP-2) leads to a rapid and dramatic fall in mitochondrial membrane potential and to a reduction of mitochondrial NADH and intracellular ATP. In HeLa cells, increased UCP-2 expression leads to a form of cell death that is not inhibited by the anti-apoptotic gene product Bcl-2 and that morphologically resembles cellular oncosis. We further describe the creation of a dominant interfering mutant of UCP-2 whose expression increases resting mitochondrial membrane potential and selectively increases the resistance to cell death following oncotic but not apoptotic stimuli. These results suggest that distinct genetic programs may regulate the cellular response to either apoptotic or oncotic stimuli. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Finkel, T (reprint author), NHLBI, Cardiol Branch, NIH, 10 Ctr Dr,Bldg 10-6N-240, Bethesda, MD 20892 USA. NR 30 TC 70 Z9 78 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27385 EP 27392 DI 10.1074/jbc.M111860200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900088 PM 12011039 ER PT J AU Varnai, P Lin, X Lee, SB Tuymetova, G Bondeva, T Spat, A Rhee, SG Hajnoczky, G Balla, T AF Varnai, P Lin, X Lee, SB Tuymetova, G Bondeva, T Spat, A Rhee, SG Hajnoczky, G Balla, T TI Inositol lipid binding and membrane localization of isolated pleckstrin homology (PH) domains - Studies on the PH domains of phospholipase C delta(1) and p130 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HETEROTRIMERIC G-PROTEINS; BETA-GAMMA-SUBUNITS; CATALYTICALLY INACTIVE PROTEIN; ADRENERGIC-RECEPTOR KINASE; BRUTONS TYROSINE KINASE; PHOSPHOINOSITIDE-BINDING; PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE; STRUCTURAL BASIS; FYVE DOMAIN; 1,4,5-TRISPHOSPHATE RECEPTOR AB The relationship between the ability of isolated pleckstrin homology (PH) domains to bind inositol lipids or soluble inositol phosphates in vitro and to localize to cellular membranes in live cells was examined by comparing the PH domains of phospholipase Cdelta(1) (PLCdelta(1)) and the recently cloned PLC-like protein p130 fused to the green fluorescent protein (GFP). The prominent membrane localization of PLCdelta(1)PH-GFP was paralleled with high affinity binding to inositol 1,4,5-trisphosphate (InsP(3)) as well as to phosphatidylinositol 4,5-bisphosphate-containing lipid vesicles or nitrocellulose membrane strips. In contrast, no membrane localization was observed with p130PH-GFP despite its InsP(3) and phosphatidylinositol 4,5-bisphosphate-binding properties being comparable with those of PLCdelta(1)PH-GFP. The N-terminal ligand binding domain of the type I InsP(3) receptor also failed to localize to the plasma membrane despite its 5-fold higher affinity to InsP(3) than the PH domains. By using a chimeric approach and cassette mutagenesis, the C-terminal alpha-helix and the short loop between the beta6-beta7 sheets of the PLCdelta(1)PH domain, in addition to its InsP(3)-binding region, were identified as critical components for membrane localization in intact cells. These data indicate that binding to the inositol phosphate head group is necessary but may not be sufficient for membrane localization of the PLCdelta(1)PH-GFP fusion protein, and motifs located within the C-terminal half of the PH domain provide auxiliary contacts with additional membrane components. C1 NICHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Thomas Jefferson Univ, Dept Pathol Anat & Cell Biol, Philadelphia, PA 19107 USA. NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. Semmelweis Univ, Sch Med, Dept Physiol, H-1444 Budapest, Hungary. Semmelweis Univ, Sch Med, Lab Cellular & Mol Physiol, H-1444 Budapest, Hungary. RP Balla, T (reprint author), NICHD, Endocrinol & Reprod Res Branch, NIH, Rm 6A35,Bldg 49,49 Convent Dr, Bethesda, MD 20892 USA. EM tambal@box-t.nih.gov OI Balla, Tamas/0000-0002-9077-3335 FU NIDDK NIH HHS [DK 51526]; NIGMS NIH HHS [GM 59419] NR 62 TC 81 Z9 83 U1 0 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27412 EP 27422 DI 10.1074/jbc.M109672200 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900091 PM 12019260 ER PT J AU Cavanaugh, AH Hirschler-Laszkiewicz, I Hu, QY Dundr, M Smink, T Misteli, T Rothblum, LI AF Cavanaugh, AH Hirschler-Laszkiewicz, I Hu, QY Dundr, M Smink, T Misteli, T Rothblum, LI TI Rrn3 phosphorylation is a regulatory checkpoint for ribosome biogenesis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID RNA-POLYMERASE-I; TRANSCRIPTION FACTOR UBF; FACTOR-TIF-IA; SACCHAROMYCES-CEREVISIAE ENCODES; GROWTH-DEPENDENT REGULATION; GENE-TRANSCRIPTION; RDNA TRANSCRIPTION; DNA-TRANSCRIPTION; FACTOR SL1; INITIATION AB Cycloheximide inhibits ribosomal DNA (rDNA) transcription in vivo. The mouse homologue of yeast Rrn3, a polymerase-associated transcription initiation factor, can complement extracts from cycloheximide-treated mammalian cells. Cycloheximide inhibits the phosphorylation of Rrn3 and causes its dissociation from RNA polymerase I. Rrn3 interacts with the rpa43 subunit of RNA polymerase 1, and treatment with cycloheximide inhibits the formation of a Rrn3.rpa43, complex in vivo. Rrn3 produced in Sf9 cells but not in bacteria interacts with rpa43 in vitro, and such interaction is dependent upon the phosphorylation state of Rrn3. Significantly, neither dephosphorylated Rrn3 nor Rrn3 produced in Escherichia coli can restore transcription by extracts from cycloheximide-treated cells. These results suggest that the phosphorylation. state of Rrn3 regulates rDNA transcription by determining the steady-state concentration of the Rrn3.RNA polymerase I complex within the nucleolus. C1 Sigfried & Janet Weis Ctr Res, Geisinger Clin, Danville, PA 17821 USA. NCI, NIH, Bethesda, MD 20892 USA. RP Rothblum, LI (reprint author), Sigfried & Janet Weis Ctr Res, Geisinger Clin, 100 N Acad Ave, Danville, PA 17821 USA. NR 58 TC 38 Z9 40 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 26 PY 2002 VL 277 IS 30 BP 27423 EP 27432 DI 10.1074/jbc.M201232200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577HR UT WOS:000177055900092 PM 12015311 ER PT J AU Zhao, CQ Xia, CW Mao, QK Forsterling, H DeRose, E Antholine, WE Subczynski, WK Petering, DH AF Zhao, CQ Xia, CW Mao, QK Forsterling, H DeRose, E Antholine, WE Subczynski, WK Petering, DH TI Structures of HO2-Co(III)bleomycin A(2) bound to d(GAGCTC)(2) and d(GGAAGCTTCC)(2): structure-reactivity relationships of Co and Fe bleomycins SO JOURNAL OF INORGANIC BIOCHEMISTRY LA English DT Article; Proceedings Paper CT 10th International Conference on Bioinorganic Chemistry CY AUG 26-31, 2001 CL FLORENCE, ITALY DE bleomycin; cobalt; DNA; iron; NMR; structure ID CENTER-DOT-BLEOMYCIN; ASCITES TUMOR-CELLS; SELECTIVE DNA CLEAVAGE; DOUBLE-STRAND BREAKAGE; IRON BLEOMYCIN; COBALT(II) BLEOMYCIN; GROWTH-INHIBITION; A2 GREEN; NMR; RECOGNITION AB HO2-Co(III)bleomycin is a model for HO2-Fe(III)bleomycin, which initiates single and double strand cleavage of DNA. In order to enlarge the understanding of its structure and reactivity, three-dimensional structures of HO2-Co(III)bleomycin bound to two DNA oligomers, d(GAGCTC)(2) (1) and d(GGAAGCTTCC)(2) (II), that have 5'-GC-3' binding sites, have been determined by nuclear magnetic resonance (NMR) methods. Besides previously recognized determinants of binding selectivity, a probable hydrogen bond was detected between the pyrimidinyl acetamido NH2 and the carbonyl of cytosine base paired to G at the recognition site. Another hydrogen bond between the NH of the dimethylsulfonium R group and N7 of guanine opposite cytosine at the GC site may contribute to specification of the pyrimidine. Substitution of G with inosine shifted HO2-Co(III)Blm A(2)-I and Fe(III)Blm-I into fast exchange on the NMR time scale, supporting the role of the 2-amino group in site specification for each molecule. The conformationally stable metal-domain linker established a close-packed adduct with the minor groove in which the hydroperoxide ligand occupies a sterically constrained pocket that is isolated from the solvent. The hydroperoxide group is directed toward one of the two cytosine H4' hydrogens but is sterically blocked from access to the other by the drug. These findings enlarge the structural understanding of selective binding of Co(III)/Fe(III)Blm species at G-pyrimidine sites. They also rationalize the instability of a number of ligands bound to Co(III)/Fe(III)Blm at specific binding sequences and the relative unreactivity of Fe(III)Blm-I with ascorbate as well as its lack of interaction. with spin labels. (C) 2002 Published by Elsevier Science Inc. C1 Univ Wisconsin, Dept Chem, Milwaukee, WI 53201 USA. NIEHS, Res Triangle Pk, NC 27709 USA. Med Coll Wisconsin, Res Inst, Milwaukee, WI 53233 USA. RP Petering, DH (reprint author), Univ Wisconsin, Dept Chem, Milwaukee, WI 53201 USA. FU NCI NIH HHS [CA-22184] NR 47 TC 21 Z9 21 U1 1 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0162-0134 J9 J INORG BIOCHEM JI J. Inorg. Biochem. PD JUL 25 PY 2002 VL 91 IS 1 BP 259 EP 268 AR PII S0162-0134(02)00420-8 DI 10.1016/S0162-0134(02)00420-8 PG 10 WC Biochemistry & Molecular Biology; Chemistry, Inorganic & Nuclear SC Biochemistry & Molecular Biology; Chemistry GA 578VK UT WOS:000177141500030 PM 12121784 ER PT J AU Cosentino, L Malling, HV Heddle, JA AF Cosentino, L Malling, HV Heddle, JA TI Response of the phi X174 am3, cs70 transgene to acute and chronic ENU exposure: implications for protocol design SO MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS LA English DT Article DE treatment protocol; transgenic mice; mutagen ID MUTATIONS IN-VIVO; SOMATIC MUTATION; MOLECULAR CHARACTERISTICS; LACZ TRANSGENE; STEM-CELLS; SHORT-TERM; MICE; MUTAGENESIS; INVIVO; LOCUS AB Studies of other transgenic assays have shown that time after treatment is a very important variable in the analysis of mutation frequencies but that eventually a plateau frequency is reached, indicating that the mutations are neutral. This neutrality is very important for the design of both experiments and testing protocols. Here we show that the phi 174 am3, cs70 transgene gives qualitatively similar results to the other transgenes studied after exposure of the mice to N-ethyl-N-nitrosourea. In the small intestine, the mutant frequency induced by an acute dose did not change significantly from 10 to 70 days post-treatment, indicating that the mutations induced are, indeed, neutral. Likewise, the mutant frequency increased linearly with duration of exposure to ENU at a constant rate. Mutant frequencies obtained were 10 times higher from the chronic exposure than produced by a nearly lethal acute dose. As in previous comparisons of a transgene and the endogenous Dlb-1 locus in the small intestine, the chronic exposure was much more effective at increasing the sensitivity of the transgene than of the endogenous gene. The Dlb-1 locus shows more complex kinetics in this strain, as in others, with mutations initially accumulating at a slower rate, indicating a differential repair of genetic damage. (C) 2002 Elsevier Science B.V. All rights reserved. C1 Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94550 USA. York Univ, Dept Biol, Toronto, ON M3J 1P3, Canada. NIEHS, Mammalian Genet Grp, Toxicol Lab, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. RP Cosentino, L (reprint author), Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, 7000 East Ave, Livermore, CA 94550 USA. NR 27 TC 1 Z9 1 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1383-5718 J9 MUTAT RES-GEN TOX EN JI Mutat. Res. Genet. Toxicol. Environ. Mutagen. PD JUL 25 PY 2002 VL 518 IS 2 BP 113 EP 121 AR PII S1383-5718(02)00078-5 DI 10.1016/S1383-5718(02)00078-5 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 581CN UT WOS:000177273700002 PM 12113762 ER PT J AU Giaever, G Chu, AM Ni, L Connelly, C Riles, L Veronneau, S Dow, S Lucau-Danila, A Anderson, K Andre, B Arkin, AP Astromoff, A El Bakkoury, M Bangham, R Benito, R Brachat, S Campanaro, S Curtiss, M Davis, K Deutschbauer, A Entian, KD Flaherty, P Foury, F Garfinkel, DJ Gerstein, M Gotte, D Guldener, U Hegemann, JH Hempel, S Herman, Z Jaramillo, DF Kelly, DE Kelly, SL Kotter, P LaBonte, D Lamb, DC Lan, N Liang, H Liao, H Liu, L Luo, CY Lussier, M Mao, R Menard, P Ooi, SL Revuelta, JL Roberts, CJ Rose, M Ross-Macdonald, P Scherens, B Schimmack, G Shafer, B Shoemaker, DD Sookhai-Mahadeo, S Storms, RK Strathern, JN Valle, G Voet, M Volckaert, G Wang, CY Ward, TR Wilhelmy, J Winzeler, EA Yang, YH Yen, G Youngman, E Yu, KX Bussey, H Boeke, JD Snyder, M Philippsen, P Davis, RW Johnston, M AF Giaever, G Chu, AM Ni, L Connelly, C Riles, L Veronneau, S Dow, S Lucau-Danila, A Anderson, K Andre, B Arkin, AP Astromoff, A El Bakkoury, M Bangham, R Benito, R Brachat, S Campanaro, S Curtiss, M Davis, K Deutschbauer, A Entian, KD Flaherty, P Foury, F Garfinkel, DJ Gerstein, M Gotte, D Guldener, U Hegemann, JH Hempel, S Herman, Z Jaramillo, DF Kelly, DE Kelly, SL Kotter, P LaBonte, D Lamb, DC Lan, N Liang, H Liao, H Liu, L Luo, CY Lussier, M Mao, R Menard, P Ooi, SL Revuelta, JL Roberts, CJ Rose, M Ross-Macdonald, P Scherens, B Schimmack, G Shafer, B Shoemaker, DD Sookhai-Mahadeo, S Storms, RK Strathern, JN Valle, G Voet, M Volckaert, G Wang, CY Ward, TR Wilhelmy, J Winzeler, EA Yang, YH Yen, G Youngman, E Yu, KX Bussey, H Boeke, JD Snyder, M Philippsen, P Davis, RW Johnston, M TI Functional profiling of the Saccharomyces cerevisiae genome SO NATURE LA English DT Article ID CAENORHABDITIS-ELEGANS; YEAST GENOME; EXPRESSION; DISRUPTION; SEQUENCES; DELETION; PROTEIN; GENES AB Determining the effect of gene deletion is a fundamental approach to understanding gene function. Conventional genetic screens exhibit biases, and genes contributing to a phenotype are often missed. We systematically constructed a nearly complete collection of gene-deletion mutants (96% of annotated open reading frames, or ORFs) of the yeast Saccharomyces cerevisiae. DNA sequences dubbed 'molecular bar codes' uniquely identify each strain, enabling their growth to be analysed in parallel and the fitness contribution of each gene to be quantitatively assessed by hybridization to high-density oligonucleotide arrays. We show that previously known and new genes are necessary for optimal growth under six well-studied conditions: high salt, sorbitol, galactose, pH 8, minimal medium and nystatin treatment. Less than 7% of genes that exhibit a significant increase in messenger RNA expression are also required for optimal growth in four of the tested conditions. Our results validate the yeast gene-deletion collection as a valuable resource for functional genomics. C1 Stanford Univ, Stanford Genome Technol Ctr, Palo Alto, CA 94304 USA. Stanford Univ, Sch Med, Dept Biochem, Stanford, CA 94305 USA. Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA. Yale Univ, Dept Mol Biophys & Biochem, New Haven, CT 06520 USA. Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA. Washington Univ, Sch Med, Dept Genet, St Louis, MO 63110 USA. McGill Univ, Dept Biol, Montreal, PQ H3A 1B1, Canada. Rosetta Inpharmat Inc, Kirkland, WA 98034 USA. Catholic Univ Louvain, FYSA, B-1348 Louvain, Belgium. Free Univ Brussels, Lab Physiol Cellulaire, Gosselies, Belgium. Univ Calif Berkeley, Dept Bioengn, Berkeley, CA 94720 USA. Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA. Univ Calif Berkeley, Lawrence Berkeley Lab, Phys Biosci Div, Howard Hughes Med Inst, Berkeley, CA 94720 USA. Free Univ Brussels, IRMW, B-1070 Brussels, Belgium. Univ Salamanca, CSIC, Inst Microbiol & Bioquim, Dept Genet & Microbiol, E-37007 Salamanca, Spain. Univ Basel, Biozentrum, Dept Mol Microbiol, CH-4056 Basel, Switzerland. Univ Padua, Dept Biol, I-35121 Padua, Italy. Goethe Univ Frankfurt, Inst Microbiol, EUROSCARF, D-60439 Frankfurt, Germany. Univ Calif Berkeley, Dept Elect Engn & Comp Sci, Berkeley, CA 94720 USA. NCI, Ctr Canc Res, Gene Regulat & Chromosome Biol Lab, Frederick, MD 21702 USA. Univ Dusseldorf, Inst Mikrobiol, D-40225 Dusseldorf, Germany. Univ Wales, Inst Biol Sci, Aberystwyth SY23 3DA, Dyfed, Wales. Concordia Univ, Dept Biol, Montreal, PQ H3G 1M8, Canada. Katholieke Univ Leuven, Lab Gene Technol, B-3001 Heverlee, Belgium. RP Davis, RW (reprint author), Stanford Univ, Stanford Genome Technol Ctr, Palo Alto, CA 94304 USA. EM dbowe@cmgm.stanford.edu RI Arkin, Adam/A-6751-2008; Flaherty, Patrick/A-1720-2011; Andre, Bruno/D-1725-2010; Entian, Karl-Dieter/D-3080-2011; Revuelta, Jose/C-2324-2012; Guldener, Ulrich/G-5227-2012; Storms, Reginald/B-2405-2013; Benito, Rocio/B-4271-2017; OI VALLE, GIORGIO/0000-0003-4377-5685; CAMPANARO, STEFANO/0000-0002-9431-1648; Arkin, Adam/0000-0002-4999-2931; Revuelta, Jose/0000-0001-7838-5308; Guldener, Ulrich/0000-0001-5052-8610; Benito, Rocio/0000-0001-9781-4198; Boeke, Jef/0000-0001-5322-4946; Johnston, Mark/0000-0002-4932-7229 NR 22 TC 2258 Z9 3440 U1 12 U2 192 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 EI 1476-4687 J9 NATURE JI Nature PD JUL 25 PY 2002 VL 418 IS 6896 BP 387 EP 391 DI 10.1038/nature00935 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 576MU UT WOS:000177009700029 PM 12140549 ER PT J AU Lauwereyns, J Watanabe, K Coe, B Hikosaka, O AF Lauwereyns, J Watanabe, K Coe, B Hikosaka, O TI A neural correlate of response bias in monkey caudate nucleus SO NATURE LA English DT Article ID SACCADIC EYE-MOVEMENTS; NEURONAL-ACTIVITY; FUNCTIONAL-PROPERTIES; BASAL GANGLIA; REWARD; EXPECTATION; CORTEX; SIGNALS; TARGET AB Primates are equipped with neural circuits in the prefrontal cortex(1-6), the parietal cortex(7) and the basal ganglia(6,8-11) that predict the availability of reward during the performance of behavioural tasks. It is not known, however, how reward value is incorporated in the control of action. Here we identify neurons in the monkey caudate nucleus that create a spatially selective response bias depending on the expected gain. In behavioural tasks, the monkey had to make a visually guided eye movement in every trial, but was rewarded for a correct response in only half of the trials. Reward availability was predictable on the basis of the spatial position of the visual target. We found that caudate neurons change their discharge rate systematically, even before the appearance of the visual target, and usually fire more when the contralateral position is associated with reward. Strong anticipatory activity of neurons with a contralateral preference is associated with decreased latency for eye movements in the contralateral direction. We conclude that this neuronal mechanism creates an advance bias that favours a spatial response when it is associated with a high reward value. C1 NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. Juntendo Univ, Dept Physiol, Bunkyo Ku, Tokyo 1130033, Japan. RP Hikosaka, O (reprint author), NEI, Sensorimotor Res Lab, NIH, Bldg 49,Room 2A50, Bethesda, MD 20892 USA. OI Lauwereyns, Jan/0000-0003-0551-2550 NR 28 TC 240 Z9 244 U1 2 U2 13 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 25 PY 2002 VL 418 IS 6896 BP 413 EP 417 DI 10.1038/nature00892 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 576MU UT WOS:000177009700037 PM 12140557 ER PT J AU Stoll, BJ Hansen, N Fanaroff, AA Wright, LL Carlo, WA Ehrenkranz, RA Lemons, JA Donovan, EF Stark, AR Tyson, JE Oh, W Bauer, CR Korones, SB Shankaran, S Laptook, AR Stevenson, DK Papile, LA Poole, WK AF Stoll, BJ Hansen, N Fanaroff, AA Wright, LL Carlo, WA Ehrenkranz, RA Lemons, JA Donovan, EF Stark, AR Tyson, JE Oh, W Bauer, CR Korones, SB Shankaran, S Laptook, AR Stevenson, DK Papile, LA Poole, WK TI Changes in pathogens causing early-onset sepsis in very-low-birth-weight infants SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID NEONATAL SEPSIS; ESCHERICHIA-COLI; INFECTION; AMPICILLIN; DISEASE; CONSEQUENCES; ANTIBIOTICS; RESISTANCE; PREVENTION; HOSPITALS AB Background It is uncertain whether the rates and causes of early-onset sepsis (that occurring within 72 hours after birth) among very-low-birth-weight infants have changed in recent years, since antibiotics have begun to be used more widely during labor and delivery. Methods We studied 5447 very-low-birth-weight infants (those weighing between 401 and 1500 g) born at centers of the Neonatal Research Network of the National Institute of Child Health and Human Development between 1998 and 2000 who had at least one blood culture in the first three days of life and compared them with 7606 very-low-birth-weight infants born at centers in the network between 1991 and 1993. Results Early-onset sepsis (as confirmed by positive blood cultures) was present in 84 infants in the more recent birth cohort (1.5 percent). As compared with the earlier birth cohort, there was a marked reduction in group B streptococcal sepsis (from 5.9 to 1.7 per 1000 live births of infants weighing 401 to 1500 g, P < 0.001) and an increase in Escherichia coli sepsis (from 3.2 to 6.8 per 1000 live births, P = 0.004); the overall rate of early-onset sepsis was not significantly changed. Most E. coli isolates from the recent birth cohort (85 percent) were resistant to ampicillin, and mothers of infants with ampicillin-resistant E. coli infections were more likely to have received intrapartum ampicillin than were those with ampicillin-sensitive strains (26 of 28 with sensitivity data vs. 1 of 5, P = 0.01). Infants with early-onset sepsis were more likely to die than uninfected infants (37 percent vs. 13 percent, P < 0.001), especially if they were infected with gram-negative organisms. Conclusions Early-onset sepsis remains an uncommon but potentially lethal problem among very-low-birth-weight infants. The; change in pathogens over time from predominantly gram-positive to predominantly gram-negative requires confirmation by ongoing surveillance. C1 Emory Univ, Sch Med, Dept Pediat, Atlanta, GA 30322 USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. Case Western Reserve Univ, Dept Pediat, Cleveland, OH 44106 USA. NICHHD, Bethesda, MD 20892 USA. Univ Alabama, Dept Pediat, Birmingham, AL USA. Yale Univ, Sch Med, Dept Pediat, New Haven, CT 06510 USA. Indiana Univ, Sch Med, Dept Pediat, Indianapolis, IN 46202 USA. Univ Cincinnati, Coll Med, Dept Pediat, Cincinnati, OH USA. Brigham & Womens Hosp, Dept Newborn Med, Boston, MA 02115 USA. Univ Texas, Houston, TX USA. Brown Univ, Women & Infants Hosp, Providence, RI USA. Univ Miami, Dept Pediat, Miami, FL 33152 USA. Univ Tennessee, Newborn Ctr, Memphis, TN USA. Wayne State Univ, Div Neonatol & Perinatal Med, Detroit, MI USA. Univ Texas, SW Med Ctr, Dept Pediat, Dallas, TX USA. Stanford Univ, Div Neonatol, Med Ctr, Palo Alto, CA 94304 USA. Univ New Mexico, Sch Med, Dept Pediat, Albuquerque, NM 87131 USA. RP Stoll, BJ (reprint author), Emory Univ, Sch Med, Dept Pediat, 2040 Ridgewood Dr NE, Atlanta, GA 30322 USA. FU NICHD NIH HHS [U10 HD27851, U01 HD36790, U10 HD21364, U10 HD21373, U10 HD21385, U10 HD21397, U10 HD21415, U10 HD27853, U10 HD27856, U10 HD27871, U10 HD27880, U10 HD27881, U10 HD27904, U10 HD34167, U10 HD34216, U10 HD40689] NR 34 TC 367 Z9 387 U1 0 U2 10 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUL 25 PY 2002 VL 347 IS 4 BP 240 EP 247 DI 10.1056/NEJMoa012657 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 587WB UT WOS:000177665800003 PM 12140299 ER PT J AU Uwaifo, GI Elberg, J Yanovski, JA AF Uwaifo, GI Elberg, J Yanovski, JA TI Impaired glucose tolerance in obese children and adolescents SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter ID INDEX C1 NIH, Bethesda, MD 20892 USA. RP Uwaifo, GI (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. RI Uwaifo, Gabriel/M-2361-2016 OI Uwaifo, Gabriel/0000-0002-6962-9304 NR 7 TC 15 Z9 15 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUL 25 PY 2002 VL 347 IS 4 BP 290 EP 291 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 587WB UT WOS:000177665800018 PM 12141328 ER PT J AU Samuni, A Goldstein, S Russo, A Mitchell, JB Krishna, MC Neta, P AF Samuni, A Goldstein, S Russo, A Mitchell, JB Krishna, MC Neta, P TI Kinetics and mechanism of hydroxyl radical and OH-adduct radical reactions with nitroxides and with their hydroxylamines SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID AROMATIC HYDROXYLATION; AQUEOUS-SOLUTIONS; TEREPHTHALATE; SUPEROXIDE; RADIOLYSIS; DOSIMETER; IRON(II); TEMPOL AB Stable nitroxide radicals are potent antioxidants and are among the most effective non-thiol radioprotectants, although they react with hydroxyl radicals more slowly than typical phenolic antioxidants or thiols. Surprisingly, the reduced forms of cyclic nitroxides, cyclic hydroxylamines, are better reductants yet have no radioprotective activity. To clarify the reason for this difference, we studied the kinetics and mechanisms of the reactions of nitroxides and their hydroxylamines with OH radicals and with OH-adducts by using pulse radiolysis, fluorimetric determination of phenolic radiation products, and electron paramagnetic resonance spectrometric determination of nitroxide concentrations following radiolysis. Competition kinetics with phenylalanine as a reference compound in pulse radiolysis experiments yielded rate constants of (4.5 +/- 0.4) x 10(9) m(-1) s(-1) for the reaction of (OH)-O-. radical with 2,2,6,6-tetramethylpiperidine-N-oxyl (TPO), 4-hydroxy-TPO (4-OH-TPO), and 4-oxo-TPO (4-O-TPO), (3.0 +/- 0.3) x 10(9) m(-1) s(-1) for deuterated 4-O-TPO, and (1.0 +/- 0.1) x 10(9) m(-1) s(-1) for the hydroxylamine 4-OH-TPO-H. The kinetic isotope effect suggests the occurrence of both (OH)-O-. addition to the aminoxyl moiety of 4-O-TPO and H-atom abstraction from the 2- or 6-methyl groups or from the 3- and 5-methylene positions. This conclusion was further supported by final product analysis, which demonstrated that (OH)-O-. partially oxidizes 4-O-TPO to the corresponding oxoammonium cation. The rate constants for the reactions of the nitroxides with the OH-adducts of phenylalanine and terephthalate have been determined to be near 4 x 10(6) m(-1) s(-1), whereas the hydroxylamine reacted at least 50 times slower, if at all. These findings indicate that the reactivity toward (OH)-O-. does not explain the differences between the radioprotective activities of nitroxides and hydroxylamines. Instead, the radioprotective activity of nitroxides, but not of hydroxylamines, can be partially attributed to their ability to detoxify OH-derived secondary radicals. C1 Hebrew Univ Jerusalem, Hadassah Med Sch, Fac Med, Dept Mol Biol, IL-91120 Jerusalem, Israel. NCI, Radiat Biol Branch, NIH, Bethesda, MD 20892 USA. Hebrew Univ Jerusalem, Dept Phys Chem, IL-91904 Jerusalem, Israel. Natl Inst Stand & Technol, Phys & Chem Properties Div, Gaithersburg, MD 20899 USA. RP Samuni, A (reprint author), Hebrew Univ Jerusalem, Hadassah Med Sch, Fac Med, Dept Mol Biol, IL-91120 Jerusalem, Israel. NR 28 TC 99 Z9 100 U1 4 U2 50 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0002-7863 J9 J AM CHEM SOC JI J. Am. Chem. Soc. PD JUL 24 PY 2002 VL 124 IS 29 BP 8719 EP 8724 AR UNSP JA017587H DI 10.1021/ja017587h PG 6 WC Chemistry, Multidisciplinary SC Chemistry GA 575EF UT WOS:000176933100044 PM 12121116 ER PT J AU Pryor, DE O'Brate, A Bilcer, G Diaz, JF Wang, YF Wang, Y Kabaki, M Jung, MK Andreu, JM Ghosh, AK Giannakakou, P Hamel, E AF Pryor, DE O'Brate, A Bilcer, G Diaz, JF Wang, YF Wang, Y Kabaki, M Jung, MK Andreu, JM Ghosh, AK Giannakakou, P Hamel, E TI The microtubule stabilizing agent laulimalide does not bind in the taxoid site, kills cells resistant to paclitaxel and epothilones, and may not require its epoxide moiety for activity SO BIOCHEMISTRY LA English DT Article ID ALPHA-BETA-TUBULIN; PURIFIED TUBULIN; SARCODICTYIN-A; CANCER-CELLS; POLYMERIZATION; DISCODERMOLIDE; ELEUTHEROBIN; PROTEINS; TAXOL(R) AB Laulimalide is a cytotoxic natural product that stabilizes microtubules. The compound enhances tubulin assembly, and laulimalide is quantitatively comparable to paclitaxel in its effects on the reaction. Laulimalide is also active in P-glycoprotein overexpressing cells, while isolaulimalide, a congener without the drug's epoxide moiety, was reported to have negligible cytotoxic and biochemical activity [Mooberry et al. (1999) Cancer Res. 59, 653-660]. We report here that laulimalide binds at a site on tubulin polymer that is distinct from the taxoid site. We found that laulimalide, while as active as paclitaxel, epothilone A, and eleutherobin in promoting the assembly of cold-stable microtubules, was unable to inhibit the binding of radiolabeled paclitaxel or of 7-O-[N-(2,7-difluoro-4'-fluoresceincarbonyl)-L-alanyl]paclitaxel, a fluorescent paclitaxel derivative, to tubulin. Confirming this observation, we demonstrated that microtubules formed in the presence of both laulimalide and paclitaxel contained near-molar quantities, relative to tubulin, of both drugs. Laulimalide was active against cell lines resistant to paclitaxel or epothilones A and B on the basis of mutations in the M40 human beta-tubulin gene. We also report that a laulimalide analogue lacking the epoxide moiety, while less active than laulimalide in biochemical and cellular systems, is probably more active than isolaulimalide. Further exploration of the role of the epoxide in the interaction of laulimalide with tubulin is therefore justified. C1 NCI, Screening Technol Branch, Dev Therapeut Program, Div Canc Treatment & Diag,NIH, Frederick, MD 21702 USA. Emory Univ, Sch Med, Winship Canc Inst, Atlanta, GA 30322 USA. Univ Illinois, Dept Chem, Chicago, IL USA. CSIC, Ctr Invest Biol, E-28006 Madrid, Spain. NCI, Sci Applicat Int Corp, NIH, Frederick, MD 21702 USA. RP Hamel, E (reprint author), NCI, Screening Technol Branch, Dev Therapeut Program, Div Canc Treatment & Diag,NIH, Bldg 469,Room 104, Frederick, MD 21702 USA. OI Andreu, Jose M/0000-0001-8064-6933; Giannakakou, Paraskevi/0000-0001-7378-262X; Diaz, J. Fernando/0000-0003-2743-3319 FU NCI NIH HHS [N01-CO-1240]; NIGMS NIH HHS [GM 55600] NR 26 TC 167 Z9 176 U1 1 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 23 PY 2002 VL 41 IS 29 BP 9109 EP 9115 AR UNSP BI020211B DI 10.1021/bi020211b PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 574NF UT WOS:000176895900013 PM 12119025 ER PT J AU Jordan, J Lipp, A Tank, J Schroder, C Stoffels, M Franke, G Diedrich, A Arnold, G Goldstein, DS Sharma, AM Luft, FC AF Jordan, J Lipp, A Tank, J Schroder, C Stoffels, M Franke, G Diedrich, A Arnold, G Goldstein, DS Sharma, AM Luft, FC TI Catechol-O-methyltransferase and blood pressure in humans SO CIRCULATION LA English DT Article DE baroreflex; nervous system, autonomic; multiple system atrophy; Shy-Drager syndrome; receptors, adrenergic ID AUTONOMIC FAILURE; PARKINSONS-DISEASE; ORTHOSTATIC HYPOTENSION; HEALTHY-VOLUNTEERS; COMT INHIBITOR; ENTACAPONE; METABOLISM; BAROREFLEX; HEMODYNAMICS; HYPERTENSION AB Background-Whether catechol-O-methyltransferase (COMT). the enzyme that metabolizes extraneuronal norepinephrine, contributes to blood pressure regulation in humans is unknown. Methods and Results-We studied incremental doses of the COMT inhibitor entacapone, the sympathetic stimulant yohimbine, and placebo in 7 patients with multiple system atrophy (Shy Drager syndrome). We selected these unique subjects because norepinephrine exerts an exaggerated increase in blood pressure in these patients. Autonomic regulation was characterized with intravenous phenylephrine, nitroprusside, and trimethaphan, Patients were extremely hypersensitive to phenylephrine and nitroprusside. Trimethaphan elicited a profound depressor response. Phenylephrine sensitivity increased only slightly during ganglionic blockade. Entacapone increased systolic blood pressure dose-dependently, however. the pressor response to yohimbine was approximate to3.5 times greater than the maximal response to entacapone. Conclusions-COMT inhibition elicits a moderate, dose-dependent pressor response in the setting of severely impaired baroreflex buffering. Patients with multiple system atrophy allow for the characterization of subtle manipulations of norepinephrine turnover and blood pressure regulation in small numbers of subjects. C1 Humboldt Univ, Franz Volhard Clin, Clin Res Ctr, Med Fac,Charite, D-13125 Berlin, Germany. Humboldt Univ, HELIOS Klinikum Berlin, Fac Med, Charite, D-13125 Berlin, Germany. Humboldt Univ, Dept Neurol, Fac Med, Charite, D-13125 Berlin, Germany. Vanderbilt Univ, Autonom Dysfunct Serv, Nashville, TN USA. Natl Inst Neurol Disorders & Stroke, Clin Neurocardiol Sect, NIH, Bethesda, MD USA. RP Jordan, J (reprint author), Humboldt Univ, Franz Volhard Clin, Clin Res Ctr, Med Fac,Charite, Wiltbergstr 50, D-13125 Berlin, Germany. OI Luft, Friedrich/0000-0002-8635-1199 NR 34 TC 14 Z9 16 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUL 23 PY 2002 VL 106 IS 4 BP 460 EP 465 DI 10.1161/01.CIR.0000022844.50161.3B PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 579AV UT WOS:000177154000014 PM 12135946 ER PT J AU Dormann, D Weijer, G Parent, CA Devreotes, PN Weijer, CJ AF Dormann, D Weijer, G Parent, CA Devreotes, PN Weijer, CJ TI Visualizing PI3 kinase-mediated cell-cell signaling during Dictyostelium development SO CURRENT BIOLOGY LA English DT Article ID DENSITY WAVE-PROPAGATION; ADENYLYL-CYCLASE; NUCLEAR TRANSLOCATION; LIVING CELLS; BETA-GAMMA; CHEMOTAXIS; DISCOIDEUM; MOVEMENT; PROTEIN; ACTIVATION AB Background: Starving amoebae of Dictyostelium discoideum communicate by relaying extracellular cAMP signals, which direct chemotactic movement, resulting in the aggregation of thousands of cells into multicellular aggregates. Both cAMP relay and chemotaxis require the activation of PI3 kinase signaling. The spatiotemporal dynamics of PI3 kinase signaling can be followed in individual cells via the cAMP-induced membrane recruitment of a GFP-tagged PH domain-containing protein, CRAC, which is required for the activation of adenylylcyclase. Results: We show that polarized periodic CRAC-GFP translocation occurs during the aggregation and mound stages of development in response to periodic cAMP signals. The duration of CRAC translocation to the membrane is determined by the duration of the rising phase of the cAMP signal. The system shows rapid adaptation and responds to the rate of change of the extracellular cAMP concentration. When the cells are in close contact, it takes 10 s for the signal to propagate from one cell to the next. In slugs, all cells show a permanent polarized PI3 kinase signaling in their leading edge, which is dependent on cell-cell contact. Conclusions: Measuring the redistribution of GFP-tagged CRAC has enabled us to study the dynamics of PI3 kinase-mediated cell-cell communication at the individual cell level in the multicellular stages of Dictyostelium development. This approach should also be useful to study the interactions between cell-cell signaling, cell polarization, and movement in the development of other organisms. C1 Univ Dundee, Sch Life Sci, Wellcome Trust Bioctr, Dundee DD1 5EH, Scotland. NCI, Cellular & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Biol Chem, Baltimore, MD 21205 USA. RP Weijer, CJ (reprint author), Univ Dundee, Sch Life Sci, Wellcome Trust Bioctr, Dow St, Dundee DD1 5EH, Scotland. RI weijer, cornelis/I-4538-2012; OI Weijer, Cornelis/0000-0003-2192-8150 NR 35 TC 47 Z9 49 U1 0 U2 0 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 23 PY 2002 VL 12 IS 14 BP 1178 EP 1188 AR PII S0960-9822(02)00950-8 DI 10.1016/S0960-9822(02)00950-8 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 577ZW UT WOS:000177093800016 PM 12176327 ER PT J AU Brzostowski, JA Johnson, C Kimmel, AR AF Brzostowski, JA Johnson, C Kimmel, AR TI G alpha-mediated inhibition of developmental signal response SO CURRENT BIOLOGY LA English DT Article ID HETEROTRIMERIC G-PROTEINS; G-BETA-GAMMA; DICTYOSTELIUM-DISCOIDEUM; CHEMOTACTIC CELLS; RECEPTORS; SUBUNIT; RGS; DIFFERENTIATION; MORPHOGENESIS; ACTIVATION AB Background: Seven-transmembrane receptor (7-TMR)-G protein networks are molecular sensors of extracellular signals in all eukarya. These pathways cycle through activated (sensitized) and inhibited (desensitized) states, and, while many of the molecular components for signal activation have been described, inhibitory mechanisms are not well characterized. In Dictyostelium, 7-TM cAMP receptors direct chemotaxis and development but also regulate the periodic synthesis of their own ligand, the chemoattractant/morphogen cAMP. We now demonstrate through loss-of-function/gain-of-function studies that the novel heterotrimeric Gag protein subunit regulates an inhibitory pathway during early Dictyostelium development for the cAMP signal response. Results: galpha9 null cells form more cAMP signaling centers, are more resistant to compounds that inhibit cAMP signaling, and complete aggregation sooner and at lower cell densities than wild-type cells. These phentoypes are consistent with the loss of an inhibitory signaling pathway during development of galpha9 null cells. Cells expressing constitutively activated Galpha9 are defective in cAMP signaling center formation and development at low cell density and display an increased sensitivity to cAMP signal inhibition that is characteristic of enhanced suppression of the cAMP signal response. Finally, we demonstrate that galpha9 null cells, which have been codeveloped with a majority of wild-type cells, primarily establish cAMP signaling centers and are able to non-autonomously direct wild-type cells to adopt a galpha9 null-like phenotype. Conclusions: We suggest that Gag functions in an inhibitory-feed back pathway that regulates cAMP signaling center formation and propagation. Galpha9 may be part of the mechanism that regulates lateral signal inhibition or that modulates receptor desensitization. C1 NIDDKD, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Kimmel, AR (reprint author), NIDDKD, Cellular & Dev Biol Lab, NIH, Bldg 50-3351, Bethesda, MD 20892 USA. NR 41 TC 32 Z9 34 U1 0 U2 1 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 23 PY 2002 VL 12 IS 14 BP 1199 EP 1208 AR PII S0960-9822(02)00953-3 DI 10.1016/S0960-9822(02)00953-3 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 577ZW UT WOS:000177093800018 PM 12176329 ER PT J AU Dasso, M AF Dasso, M TI The Ran GTPase: Theme and variations SO CURRENT BIOLOGY LA English DT Review ID NUCLEAR-PORE COMPLEX; ACTIVATING PROTEIN RANGAP1; BINDING-PROTEIN; MITOTIC-SPINDLE; IMPORTIN-BETA; TRANSPORT; EXPORT; RCC1; REGULATOR; ORGANIZATION C1 NICHD, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA. RP Dasso, M (reprint author), NICHD, Lab Gene Regulat & Dev, NIH, Bldg 18T,Room 106,18 Lib Dr, Bethesda, MD 20892 USA. OI Dasso, Mary/0000-0002-5410-1371 NR 48 TC 85 Z9 88 U1 0 U2 4 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE, CAMBRIDGE, MA 02138 USA SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 23 PY 2002 VL 12 IS 14 BP R502 EP R508 AR PII S0960-9822(02)00970-3 DI 10.1016/S0960-9822(02)00970-3 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 577ZW UT WOS:000177093800014 PM 12176353 ER PT J AU Sparks, BF Friedman, SD Shaw, DW Aylward, EH Echelard, D Artru, AA Maravilla, KR Giedd, JN Munson, J Dawson, G Dager, SR AF Sparks, BF Friedman, SD Shaw, DW Aylward, EH Echelard, D Artru, AA Maravilla, KR Giedd, JN Munson, J Dawson, G Dager, SR TI Brain structural abnormalities in young children with autism spectrum disorder SO NEUROLOGY LA English DT Article ID DIAGNOSTIC OBSERVATION SCHEDULE; EARLY INFANTILE-AUTISM; CHILDHOOD AUTISM; MRI; HIPPOCAMPUS; AMYGDALA; VOLUME; AREA; INDIVIDUALS; CEREBELLAR AB Objective: To explore the specific gross neuroanatomic substrates of this brain developmental disorder, the authors examine brain morphometric features in a large sample of carefully diagnosed 3- to 4-year-old children with autism spectrum disorder (ASD) compared with age-matched control groups of typically developing (TD) children and developmentally delayed (DD) children. Methods: Volumes of the cerebrum, cerebellum, amygdala, and hippocampus were measured from three-dimensional coronal MR images acquired from 45 children with ASD, 26 TD children, and 14 DD children. The volumes were analyzed with respect to age, sex, volume of the cerebrum, and clinical status. Results: Children with ASD were found to have significantly increased cerebral volumes compared with TD and DD children. Cerebellar volume for the ASD group was increased in comparison with the TD group, but this increase was proportional to overall increases in cerebral volume. The DD group had smaller cerebellar volumes compared with both of the other groups. Measurements of amygdalae and hippocampi in this group of young children with ASD revealed enlargement bilaterally that was proportional to overall increases in total cerebral volume. There were similar findings of cerebral enlargement for both girls and boys with ASD. For subregion analyses, structural abnormalities were observed primarily in boys, although this may reflect low statistical power issues because of the small sample (seven girls with ASD) studied. Among the ASD group, structural findings were independent of nonverbal IQ. In a subgroup of children with ASD with strictly defined autism, amygdalar enlargement was in excess of increased cerebral volume. Conclusions: These structural findings suggest abnormal brain developmental processes early in the clinical course of autism. Research currently is underway to better elucidate mechanisms underlying these structural abnormalities and their longitudinal progression. C1 Univ Washington, Sch Med, Dept Radiol, Seattle, WA 98105 USA. Univ Washington, Sch Med, Dept Psychiat, Seattle, WA 98105 USA. Univ Washington, Sch Med, Dept Bioengn, Seattle, WA 98105 USA. NIMH, Bethesda, MD 20892 USA. Univ Washington, Sch Med, Ctr Human Dev & Disabil, Seattle, WA 98105 USA. RP Dager, SR (reprint author), Univ Washington, Sch Med, Dept Radiol, 4225 Roosevelt Way NE,Suite 306-C, Seattle, WA 98105 USA. RI Giedd, Jay/A-3080-2008; Giedd, Jay/B-7302-2012; Giedd, Jay/J-9644-2015 OI Giedd, Jay/0000-0003-0827-3460; Giedd, Jay/0000-0003-2002-8978 FU NICHD NIH HHS [P01 HD34565] NR 48 TC 455 Z9 468 U1 19 U2 77 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUL 23 PY 2002 VL 59 IS 2 BP 184 EP 192 PG 9 WC Clinical Neurology SC Neurosciences & Neurology GA 574EL UT WOS:000176875400008 PM 12136055 ER PT J AU Gaillard, WD Balsamo, L Xu, B Grandin, CB Braniecki, SH Papero, PH Weinstein, S Conry, J Pearl, PL Sachs, B Sato, S Jabbari, B Vezina, LG Frattali, C Theodore, WH AF Gaillard, WD Balsamo, L Xu, B Grandin, CB Braniecki, SH Papero, PH Weinstein, S Conry, J Pearl, PL Sachs, B Sato, S Jabbari, B Vezina, LG Frattali, C Theodore, WH TI Language dominance in partial epilepsy patients identified with an fMRI reading task SO NEUROLOGY LA English DT Article ID POSITRON-EMISSION-TOMOGRAPHY; INFERIOR PREFRONTAL CORTEX; CEREBRAL BLOOD-FLOW; FUNCTIONAL MRI; RIGHT-HEMISPHERE; WADA TEST; ELECTROCORTICAL STIMULATION; CORTICAL REPRESENTATION; SENTENCE COMPREHENSION; AUDITORY COMPREHENSION AB Background: fMRI language tasks readily identify frontal language areas; temporal activation has been less consistent. No studies have compared clinical visual judgment to quantitative region of interest (ROI) analysis. Objective: To identify temporal language areas in patients with partial epilepsy using a reading paradigm with clinical and ROI interpretation. Methods: Thirty patients with temporal lobe epilepsy, aged 8 to 56 years, had 1.5-T fMRI. Patients silently named an object described by a sentence compared to a visual control. Data were analyzed with ROI analysis from t-maps. Regional asymmetry indices (AI) were calculated ([L-R]/[L+R]) and language dominance defined as >0.20. t-Maps were visually rated by three readers at three t thresholds. Twenty-one patients had intracarotid amobarbital test (IAT). Results: The fMRI reading task provided evidence of language lateralization in 27 of 30 patients with ROI analysis. Twenty-five were left dominant, two right, one bilateral, and two were nondiagnostic; IAT and fMRI agreed in most patients, three had partial agreement, none overtly disagreed. Interrater agreement ranged between 0.77 to 0.82 (Cramer V; p < 0.0001); agreement between visual and ROI reading with IAT was 0.71 to 0.77 (Cramer V; p < 0.0001). Viewing data at lower thresholds added interpretation to 12 patients on visual analysis and 8 with ROI analysis. Conclusions: An fMRI reading paradigm can identify language dominance in frontal and temporal areas. Clinical visual interpretation is comparable to quantitative ROI analysis. C1 George Washington Univ, Sch Med, Childrens Natl Med Ctr, Dept Neurosci, Washington, DC USA. George Washington Univ, Sch Med, Childrens Natl Med Ctr, Dept Radiol, Washington, DC USA. Natl Inst Neurol Disorders & Stroke, Epilepsy Res Branch, Bethesda, MD USA. NIH, Speech Language Pathol Sect, Dept Rehabil Med, Ctr Clin, Bethesda, MD USA. Uniformed Serv Univ Hlth Sci, Dept Neurol, Bethesda, MD USA. Univ Catholique Louvain, St Luc Univ Hosp, Dept Radiol, Brussels, Belgium. RP Gaillard, WD (reprint author), Childrens Natl Med Ctr, Dept Neurol, 111 Michigan Ave NW, Washington, DC 20010 USA. FU NINDS NIH HHS [KO8-NS1663] NR 62 TC 144 Z9 147 U1 9 U2 13 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUL 23 PY 2002 VL 59 IS 2 BP 256 EP 265 PG 10 WC Clinical Neurology SC Neurosciences & Neurology GA 574EL UT WOS:000176875400020 PM 12136067 ER PT J AU Nestorovich, EM Danelon, C Winterhalter, M Bezrukov, SM AF Nestorovich, EM Danelon, C Winterhalter, M Bezrukov, SM TI Designed to penetrate: Time-resolved interaction of single antibiotic molecules with bacterial pores SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ESCHERICHIA-COLI; OUTER-MEMBRANE; BETA-LACTAMS; DRUG-RESISTANCE; PORIN CHANNELS; TRANSPORT; DIFFUSION; PROTEIN; PERMEABILITY; MECHANISM AB Membrane permeability barriers are among the factors contributing to the intrinsic resistance of bacteria to antibiotics. We have been able to resolve single ampicillin molecules moving through a channel of the general bacterial porin, OmpF (outer membrane protein F), believed to be the principal pathway for the beta-lactam antibiotics. With ion channel reconstitution and high-resolution conductance recording, we find that ampicillin and several other efficient penicillins and cephalosporins strongly interact with the residues of the constriction zone of the OmpF channel. Therefore, we hypothesize that, in analogy to substrate-specific channels that evolved to bind certain metabolite molecules, antibiotics have "evolved" to be channel-specific. Molecular modeling suggests that the charge distribution of the ampicillin molecule complements the charge distribution at the narrowest part of the bacterial porin. Interaction of these charges creates a region of attraction inside the channel that facilitates drug translocation through the constriction zone and results in higher permeability rates. C1 NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. Inst Pharmacol & Biol Struct, F-31077 Toulouse, France. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. RP Bezrukov, SM (reprint author), NICHHD, Lab Phys & Struct Biol, NIH, Bldg 9,Room 1E-122, Bethesda, MD 20892 USA. NR 33 TC 175 Z9 180 U1 0 U2 15 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 9789 EP 9794 DI 10.1073/pnas.152206799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400036 PM 12119404 ER PT J AU Ellis, CA Vos, MD Howell, H Vallecorsa, T Fults, DW Clark, GJ AF Ellis, CA Vos, MD Howell, H Vallecorsa, T Fults, DW Clark, GJ TI Rig is a novel Ras-related protein and potential neural tumor suppressor SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID FARNESYLTRANSFERASE INHIBITORS; FAMILY PROTEINS; GENE; TRANSFORMATION; HETEROZYGOSITY; EXPRESSION; CELLS; FARNESYLATION; ASTROCYTOMAS; CARCINOMAS AB The Ras superfamily consists of a large group of monomeric GTPases demonstrating homology to Ras oncoproteins. Although structurally similar, Ras-superfamily proteins are functionally diverse. Whereas some members exhibit oncogenic properties, others may serve as tumor suppressors. We have identified a novel Ras-related protein that suppresses cell growth and have designated it Rig (Ras-related inhibitor of cell growth). Overexpression of Rig inhibited Ras-mediated cellular transformation and activation of downstream signaling in NIH 3T3 cells. rig mRNA is expressed at high levels in normal cardiac and neural tissue. However, Rig protein expression is frequently lost or down-regulated in neural tumor-derived cell lines and primary human neural tumors. Moreover, expression of exogenous Rig in human astrocytoma cells suppressed growth. Rig has a C-terminal CAAX motif that codes for posttranslational modification by both farnesyl and geranylgeranyl isoprenoid lipids. Consequently, Rig may play a role in the cellular response to farnesyl transferase inhibitors. Rig bears 63% overall sequence homology to a recently described Ras-family member Noey2, a tumor suppressor in breast and ovarian tissue. Therefore, Rig and Noey2 may represent a new subfamily of Ras-like tumor suppressors. C1 NCI, Dept Cell & Canc Biol, NIH, Rockville, MD 20850 USA. Univ Utah, Sch Med, Dept Neurosurg, Salt Lake City, UT 84132 USA. Huntsman Canc Inst, Salt Lake City, UT 84132 USA. RP Clark, GJ (reprint author), NCI, Dept Cell & Canc Biol, NIH, 9610 Med Ctr Dr,Suite 307, Rockville, MD 20850 USA. NR 45 TC 31 Z9 34 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 9876 EP 9881 DI 10.1073/pnas.142193799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400051 PM 12107278 ER PT J AU Eissenberg, JC Ma, JY Gerber, MA Christensen, A Kennison, JA Shilatifard, A AF Eissenberg, JC Ma, JY Gerber, MA Christensen, A Kennison, JA Shilatifard, A TI dELL is an essential RNA polymerase II elongation factor with a general role in development SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DROSOPHILA EYE DEVELOPMENT; TRANSCRIPTIONAL ELONGATION; WING MARGIN; FACTOR-SIII; ELL; PROTEIN; IDENTIFICATION; CLONING; SUPPRESSOR; EXPRESSION AB Several eukaryotic proteins increase RNA polymerase II (Pol II) transcription rates in vitro. The relative contributions of these factors to gene expression in vivo is unknown. The ELL family of proteins promote Pol II elongation in vitro, and the Drosophila ELL homolog (dELL) is associated with Pol II at sites of transcription in vivo. The purpose of this study was to test whether an ELL family protein is required for gene expression in vivo. We show that dELL is encoded by the Suppressor of Triplo-lethal locus [Su(Tpl)]. We have characterized seven distinct mutant alleles of Su(Tpl) and show that a dELL transgene rescues recessive lethality of Su(Tpl). Su(Tpl) mutations cause abnormal embryonic segmentation and dominantly modify expression of diverse genes during development. These data show that an ELL family elongation factor is essential, acts broadly in development, and is not functionally redundant to other elongation factors in vivo. C1 St Louis Univ, Sch Med, Dept Biochem & Mol Biol, St Louis, MO 63104 USA. Univ Nebraska, Sch Biol Sci, Lincoln, NE 68588 USA. NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. RP Eissenberg, JC (reprint author), St Louis Univ, Sch Med, Dept Biochem & Mol Biol, 1402 S Grand Blvd, St Louis, MO 63104 USA. FU NCI NIH HHS [R01 CA089455, 1R01CA089455]; NIGMS NIH HHS [1R55GM057005] NR 35 TC 49 Z9 50 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 9894 EP 9899 DI 10.1073/pnas.152193699 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400054 PM 12096188 ER PT J AU Jung, GM Jones, G Masison, DC AF Jung, GM Jones, G Masison, DC TI Amino acid residue 184 of yeast Hsp104 chaperone is critical for prion-curing by guanidine, prion propagation, and thermotolerance SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID SACCHAROMYCES-CEREVISIAE; TRANSLATION TERMINATION; PROTEIN HSP104; URE3 PRION; PSI+ PRION; HYDROCHLORIDE; MECHANISM; SYSTEM; STRESS; HSP70 AB inactivation of Hsp104 by guanidine is contended to be the mechanism by which guanidine cures yeast prions. We now find an Hsp104 mutation (D184N) that confers resistance to guanidine-curing of the yeast [PSI+] prion. In an independent screen we isolated an HSP104 allele altered in the same residue (D184Y) that dramatically impairs [PSI+] propagation in a temperature-dependent manner. Directed mutagenesis of HSP104 produced additional alleles that conferred varying degrees of resistance to guanidine-curing or impaired [PSI+] propagation. The mutations similarly affected propagation of the [URE3] prion. Basal and induced abundance of all mutant proteins was normal. Thermotolerance of cells expressing mutant proteins was variably resistant to guanidine, and the degree of thermotolerance did not correlate with [PSI+] stability. We thus show that guanidine cures yeast prions by inactivating Hsp104 and identify a highly conserved Hsp104 residue that is critical for yeast prion propagation. Our data suggest that Hsp104 activity can be reduced substantially without affecting [PSI+] stability, and that Hsp104 interacts differently with prion aggregates than with aggregates of thermally denatured protein. C1 NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP NIDDKD, Lab Biochem & Genet, NIH, Bldg 8,Room 407, Bethesda, MD 20892 USA. EM masisond@helix.nih.gov NR 45 TC 90 Z9 92 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 9936 EP 9941 DI 10.1073/pnas.152333299 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400062 PM 12105276 ER PT J AU Pavlov, YI Rogozin, IB Galkin, AP Aksenova, AY Hanaoka, F Rada, C Kunkel, TA AF Pavlov, YI Rogozin, IB Galkin, AP Aksenova, AY Hanaoka, F Rada, C Kunkel, TA TI Correlation of somatic hypermutation specificity and A-T base pair substitution errors by DNA polymerase eta during copying of a mouse immunoglobulin kappa light chain transgene SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID CYTIDINE DEAMINASE AID; DEPENDENT REPLICATION; TRANSLESION SYNTHESIS; PASSENGER TRANSGENES; ANTIBODY DIVERSITY; MUTATION SPECTRA; FIDELITY; GENES; SEQUENCES; CONSEQUENCES AB To test the hypothesis that inaccurate DNA synthesis by mammalian DNA polymerase eta (pol eta) contributes to somatic hypermutation (SHIM) of Ig genes, we measured the error specificity of mouse pol eta during synthesis of each strand of a mouse Ig K light chain transgene. We then compared the results to the base substitution specificity of SHM of this same gene in the mouse. The in vitro and in vivo base substitution spectra shared a number of common features. A highly significant correlation was observed for overall substitutions at A-T pairs but not for substitutions at G-C pairs. Sixteen mutational hotspots at A-T pairs observed in vivo were also found in spectra generated by mouse pol eta in vitro. The correlation was strongest for errors made by pol eta during synthesis of the non-transcribed strand, but it was also observed for synthesis of the transcribed strand. These facts, and the distribution of substitutions generated in vivo, support the hypothesis that pol eta contributes to SHM of Ig genes at A-T pairs via short patches of low fidelity DNA synthesis of both strands, but with a preference for the non-transcribed strand. C1 NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. Russian Acad Sci, Inst Cytol & Genet, Siberian Branch, Novosibirsk 630090, Russia. Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. St Petersburg State Univ, Dept Genet, St Petersburg 199034, Russia. Osaka Univ, Inst Mol & Cellular Biol, Suita, Osaka 5650871, Japan. Japan Sci & Technol Corp, CREST, Suita, Osaka 5650871, Japan. RIKEN, Inst Phys & Chem Res, Wako, Saitama 3510198, Japan. MRC, Mol Biol Lab, Cambridge CB2 2QH, England. RP Pavlov, YI (reprint author), NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. RI Galkin, Alexey/K-2968-2013; Aksenova, Anna/O-8309-2015 OI Galkin, Alexey/0000-0002-7362-8857; Aksenova, Anna/0000-0002-1601-1615 NR 47 TC 91 Z9 93 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 9954 EP 9959 DI 10.1073/pnas.152126799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400065 PM 12119399 ER PT J AU An, P Nelson, GW Wang, LH Donfield, S Goedert, JJ Phair, J Vlahov, D Buchbinder, S Farrar, WL Modi, W O'Brien, SJ Winkler, CA AF An, P Nelson, GW Wang, LH Donfield, S Goedert, JJ Phair, J Vlahov, D Buchbinder, S Farrar, WL Modi, W O'Brien, SJ Winkler, CA TI Modulating influence on HIV/AIDS by interacting RANTES gene variants SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; MULTICENTER AIDS COHORT; HIV-1 INFECTION; DISEASE PROGRESSION; LONG-TERM; PROMOTER POLYMORPHISM; CHEMOKINE RANTES; IN-VITRO; CCR5; MIP-1-ALPHA AB RANTES (regulated on activation normal T cell expressed and secreted), a ligand for the CC chemokine receptor 5, potently inhibits HIV-1 replication in vitro. We tested the influence of four RANTES single nucleotide polymorphism (SNP) variants and their haplotypes on HIV-1 infection and AIDS progression in five AIDS cohorts. Three SNPs in the RANTES gene region on chromosome 17 (403A in the promoter, In1.1C in the first intron, and 3'222C in the 3' untranslated region) are associated with increased frequency of HIV-1 infection. The common In1.1C SNP allele is nested within an intronic regulatory sequence element that exhibits differential allele binding to nuclear proteins and a down-regulation of gene transcription. The In1.1C allele or haplotypes that include In1.1C display a strong dominant association with rapid progression to AIDS among HIV-1-infected individuals in African-American, European-American, and combined cohorts. The principal RANTES SNP genetic influence on AIDS progression derives from the down-regulating RANTES In1.1C allele, although linkage disequilibrium with adjoining RANTES SNPs including a weaker up-regulating RANTES promoter allele (-28G), can modify the observed epidemiological patterns. The In1.1C-bearing genotypes account for 37% of the attributable risk for rapid progression among African Americans and may also be an important influence on AIDS progression in Africa. The diminished transcription of RANTES afforded by the In1.1C regulatory allele is consistent with increased HIV-1 spread in vivo, leading to accelerated progression to AIDS. C1 Natl Canc Inst, SAIC Frederick, Intramural Res Support Program, Ft Detrick, MD 21702 USA. NCI, Lab Genom Divers, Ft Detrick, MD 21702 USA. NCI, Lab Mol Immunoregulat, Ft Detrick, MD 21702 USA. Rho Inc, Chapel Hill, NC 27514 USA. NCI, Viral Epidemiol Branch, Bethesda, MD 20892 USA. Northwestern Univ, Sch Med, Comprehens AIDS Ctr, Chicago, IL 60611 USA. Johns Hopkins Bloomberg Sch Publ Hlth, Dept Epidemiol, Baltimore, MD 21205 USA. San Francisco Dept Publ Hlth, San Francisco, CA 94102 USA. RP Winkler, CA (reprint author), Natl Canc Inst, SAIC Frederick, Intramural Res Support Program, Ft Detrick, MD 21702 USA. FU NCI NIH HHS [N01-CO-12400, N01CO12400] NR 41 TC 138 Z9 151 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 10002 EP 10007 DI 10.1073/pnas.142313799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400073 PM 12114533 ER PT J AU Gamain, B Gratepanche, S Miller, LH Baruch, DI AF Gamain, B Gratepanche, S Miller, LH Baruch, DI TI Molecular basis for the dichotomy in Plasmodium falciparum adhesion to CD36 and chondroitin sulfate A SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ERYTHROCYTE-MEMBRANE PROTEIN-1; INFECTED ERYTHROCYTES; PARASITE ADHESION; BINDING DOMAIN; PLACENTAL MALARIA; VARIANT ANTIGEN; PREGNANT-WOMEN; CELL-SURFACE; RECEPTOR; ADHERENCE AB Plasmodium falciparum-infected erythrocytes adhere dichotomously to the host receptors CD36 and chondroitin sulfate A (CSA). This dichotomy is associated with parasite sequestration to microvasculature beds (CD36) or placenta (CSA), leading to site-specific pathogenesis. Both properties are mediated by members of the variant A falciparum erythrocyte membrane protein 1 (PfEMP-1) family and reside on nonoverlapping domains of the molecule. To identify the molecular basis for the apparent dichotomy, we expressed various domains of PfEMP-1 individually or in combination and tested their binding properties. We found that the CD36-binding mode of the cysteine-rich interdomain region-1 (CIDR1) ablates the ability of the Duffy binding-like gamma domain to bind CSA. In contrast, neither a non-CD36-binding CIDR1 nor an intercellular adhesion molecule 1 binding domain had any affect on CSA binding. Our findings point out that interactions between different domains of PfEMP-1 can alter the adhesion phenotype of infected erythrocytes and provide a molecular basis for the apparent dichotomy in adhesion. We suggest that the basis for the dichotomy is structural and that mutually exclusive conformations of PfEMP-1 are involved in binding to CD36 or CSA. Furthermore, we propose a model explaining the requirement for structural dichotomy between placental and nonplacental isolates. C1 Natl Inst Allergy & Infect Dis, Lab Parasit Dis, NIH, Bethesda, MD 20892 USA. RP Gamain, B (reprint author), Natl Inst Allergy & Infect Dis, Lab Parasit Dis, NIH, 4 Ctr Dr,Bldg 4,Rm B1-37,MSC 0425, Bethesda, MD 20892 USA. OI Gamain, Benoit/0000-0002-8255-2145 NR 31 TC 41 Z9 41 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 10020 EP 10024 DI 10.1073/pnas.152321599 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400076 PM 12096191 ER PT J AU Beres, SB Sylva, GL Barbian, KD Lei, BF Hoff, JS Mammarella, ND Liu, MY Smoot, JC Porcella, SF Parkins, LD Campbell, DS Smith, TM McCormick, JK Leung, DYM Schlievert, PM Musser, JM AF Beres, SB Sylva, GL Barbian, KD Lei, BF Hoff, JS Mammarella, ND Liu, MY Smoot, JC Porcella, SF Parkins, LD Campbell, DS Smith, TM McCormick, JK Leung, DYM Schlievert, PM Musser, JM TI Genome sequence of a serotype M3 strain of group A Streptococcus: Phage-encoded toxins, the high-virulence phenotype, and clone emergence SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE superantigen; streptococcal pyrogenic exotoxin; comparative genomics ID GROUP-A STREPTOCOCCUS; POPULATION GENETIC-ANALYSIS; GRAM-POSITIVE BACTERIA; PYROGENIC EXOTOXIN; SHOCK-SYNDROME; MOLECULAR CHARACTERIZATION; PHYLOGENETIC DISTRIBUTION; ALLELIC VARIANTS; PYOGENES; IDENTIFICATION AB Genome sequences are available for many bacterial strains, but there has been little progress in using these data to understand the molecular basis of pathogen emergence and differences in strain virulence. Serotype M3 strains of group A Streptococcus (GAS) are a common cause of severe invasive infections with unusually high rates of morbidity and mortality. To gain insight into the molecular basis of this high-virulence phenotype, we sequenced the genome of strain MGAS315, an organism isolated from a patient with streptococcal toxic shock syndrome. The genome is composed of 1,900,521 bp, and it shares approximate to1.7 Mb of related genetic material with genomes of serotype M1 and M18 strains. Phage-like elements account for the great majority of variation in gene content relative to the sequenced M1 and M18 strains. Recombination produces chimeric phages and strains with previously uncharacterized arrays of virulence factor genes. Strain MGAS315 has phage genes that encode proteins likely to contribute to pathogenesis, such as streptococcal pyrogenic exotoxin A (SpeA) and SpeK, streptococcal superantigen (SSA), and a previously uncharacterized phospholipase A(2) (designated Sla). Infected humans had anti-SpeK, -SSA, and -Sla antibodies, indicating that these GAS proteins are made in vivo. SpeK and SSA were pyrogenic and toxic for rabbits. Serotype M3 strains with the phage-encoded speK and sla genes increased dramatically in frequency late in the 20th century, commensurate with the rise in invasive disease caused by M3 organisms. Taken together, the results show that phage-mediated recombination has played a critical role in the emergence of a new, unusually virulent clone of serotype M3 GAS. C1 NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. Geospiza Inc, Seattle, WA 98107 USA. Univ Minnesota, Dept Microbiol, Sch Med, Fairview Univ Med Ctr, Minneapolis, MN 55455 USA. Natl Jewish Med & Res Ctr, Div Pediat Allergy & Immunol, Denver, CO 80262 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. RI McCormick, John/E-4232-2015; OI McCormick, John/0000-0003-3742-7289 NR 48 TC 325 Z9 755 U1 2 U2 10 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 10078 EP 10083 DI 10.1073/pnas.152298499 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400086 PM 12122206 ER PT J AU Zhang, WH Svarovskaia, ES Barr, R Pathak, VK AF Zhang, WH Svarovskaia, ES Barr, R Pathak, VK TI Y586F mutation in murine leukemia virus reverse transcriptase decreases fidelity of DNA synthesis in regions associated with adenine-thymine tracts SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID RETROVIRAL SHUTTLE VECTOR; SINGLE REPLICATION CYCLE; DOUBLE-STRANDED DNA; RNASE-H; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; TEMPLATE-PRIMER; POLYMERASE-BETA; BROAD-SPECTRUM; MINOR-GROOVE AB Using in vivo fidelity assays in which bacteria[)beta-galactosidase or green fluorescent protein genes served as reporters of mutations, we have identified a murine leukemia virus (MLV) RNase H mutant (Y586F) that exhibited an increase in the retroviral mutation rate approximate to5-fold in a single replication cycle. DNA-sequencing analysis indicated that the Y586F mutation increased the frequency of substitution mutations 17-fold within 18 nt of adenine-thymine tracts (AAAA, TTTT, or AATT), which are known to induce DNA bending. Sequence alignments indicate that MLV Y586 is equivalent to HIV-1 Y501, a component of the recently described RNase H primer grip domain, which contacts and positions the DNA primer strand near the RNase H active site. The results suggest that wild-type reverse transcriptase (RT) facilitates a specific conformation of the template-primer duplex at the polymerase active site that is important for accuracy of DNA synthesis; when an adeninethymine tract is within 18 nt of the polymerase active site, the Y586F mutant RT cannot facilitate this specific template-primer conformation, leading to an increase in the frequency of substitution mutations. These findings indicate that the RNase H primer grip can affect the template-primer conformation at the polymerase active site and that the MLV Y586 residue and template-primer conformation are important determinants of RT fidelity. C1 NCI, HIV Drug Resistance Program, Frederick, MD 21702 USA. W Virginia Univ, Dept Biol, Morgantown, WV 26506 USA. RP Pathak, VK (reprint author), NCI, HIV Drug Resistance Program, Bldg 535,Room 334, Frederick, MD 21702 USA. NR 49 TC 23 Z9 23 U1 1 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 23 PY 2002 VL 99 IS 15 BP 10090 EP 10095 DI 10.1073/pnas.152186199 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 577DG UT WOS:000177042400088 PM 12119402 ER PT J AU He, J Ogden, LG Bazzano, LA Vupputuri, S Loria, C Whelton, PK AF He, J Ogden, LG Bazzano, LA Vupputuri, S Loria, C Whelton, PK TI Dietary sodium intake and incidence of congestive heart failure in overweight US men and women - First National Health and Nutrition Examination Survey Epidemiologic Follow-up Study SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID BLOOD-PRESSURE; SALT INTAKE; HYPERTENSION; OBESITY; DISEASE; RISK; PATHOGENESIS; INFORMATION; EXCRETION; MORTALITY AB Background: Cross-sectional epidemiologic studies suggest that a higher intake of dietary sodium is associated with an increased risk of left ventricular hypertrophy. We studied the relationship between dietary sodium intake and incidence of congestive heart failure (CHF) in the first National Health and Nutrition Examination Survey Epidemiologic Follow-up Study participants. Participants and Methods: The study sample consisted of 5233 nonoverweight and 5129 overweight men and women without a history of CHF at their baseline examination. Dietary sodium and other nutrient intake estimates were obtained by a 24-hour dietary recall method at the baseline examination, conducted from 1971 to 1975. The incidence of CHF was assessed using medical records and death certificates obtained in 1982 to 1984, 1986, 1987, and 1992. Results: During an average og 19 years of follow-up we documented 413 case of CHF in nonoverweight and 679 cases of CHF in overweight participants. After adjustment for known CHF risk factors, the relative risk of CHF among overweight participants was 1.43 (95% confidence interval, 1.07-1.91) for those whose sodium intake was greater than 113.6 mmol/d compared with those whose intake was less than 50.2 mmol/d. The relative risks of CHF for a 100-mmol/d higher intake of sodium or per 1743 kcal (average energy intake in the study population) were 1.26 (95% confidence interval, 1.03-1.53) and 1.21 (95% confidence interval, 1.04-1.40), respectively. Conclusions: A higher intake of dietary sodium is a strong independent risk factor for CHF in overweight persons. A reduction in sodium intake may play an important role in the prevention of CHF in overweight individuals and populations. C1 Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, New Orleans, LA 70112 USA. Tulane Univ, Sch Publ Hlth & Trop Med, Dept Biostat, New Orleans, LA 70112 USA. NHLBI, Bethesda, MD 20892 USA. RP He, J (reprint author), Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, 1430 Tulane Ave,Mail Box SL18, New Orleans, LA 70112 USA. FU NHLBI NIH HHS [R01 HL60300, R03 HL61954] NR 39 TC 81 Z9 84 U1 0 U2 5 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD JUL 22 PY 2002 VL 162 IS 14 BP 1619 EP 1624 DI 10.1001/archinte.162.14.1619 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 573GW UT WOS:000176821800010 PM 12123406 ER PT J AU Wright, JT Agodoa, L Contreras, G Greene, T Douglas, JG Lash, J Randall, O Rogers, N Smith, MC Massry, S AF Wright, JT Agodoa, L Contreras, G Greene, T Douglas, JG Lash, J Randall, O Rogers, N Smith, MC Massry, S CA African Amer Study Kidney Dis Hy TI Successful blood pressure control in the African American study of kidney disease and hypertension SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID CHRONIC RENAL-DISEASE; CARDIOVASCULAR-DISEASE; POPULATION; TRIAL AB Background: The African American Study of Kidney Disease and Hypertension (AASK) is an ongoing trial to evaluate the effect of blood pressure and choice of antihypertensive drug on the rate of decline of renal function. Objective: To present the success of the AASK in achieving the trial's rigorous blood pressure goals in an extremely challenging patient population. Methods: The AASK participants included African American patients with hypertension (n=1094), aged 18 to 70 years, with glomerular filtration rates between 20 and 65 mL/min per 1.73 m(2) and no other identified causes of renal insufficiency. Participants were randomized to a goal mean arterial blood pressure (MAP) of either 102 to 107 mm Hg (usual MAP goal) or 92 mm Hg or less (low MAP goal). Participants in each of these groups were also randomized (double-blind) to a regimen containing metoprolol succinate, ramipril, or amlodipine besylate. Additional agents were added, if required, in the following recommended order: furosemide, doxazosin mesylate, clonidine hydrochloride, or hydralazine hydrochloride (or minoxidil, if needed). Results: In participants randomized to the low MAP goal, the percentage of participants who achieved a blood pressure of less than 140/90 mm Hg increased from a baseline of 20.0% to 78.9% by 14 months after randomization. For usual MAP goal participants, the corresponding percentages increased from 21.5% to 41.8%. The difference in median levels of MAP between the 2 MAP goal groups increased and remained at approximately 12 mm Hg. Blood pressure reduction was similar regardless of age, sex, body mass index, education, insurance or employment status, income, or marital status. Conclusion: The blood pressure goals set and achieved in AASK participants clearly demonstrate that adequate blood pressure control can be achieved even in hypertensive populations whose blood pressure is the most difficult to control. C1 Case Western Reserve Univ, Dept Med, Div Hypertens, Cleveland, OH 44106 USA. Case Western Reserve Univ, Div Nephrol, Cleveland, OH 44106 USA. Univ So Calif, Dept Med, Div Nephrol, Los Angeles, CA USA. Emory Univ, Dept Med, Div Hypertens, Atlanta, GA 30322 USA. Howard Univ, Dept Med, Div Cardiol, Washington, DC 20059 USA. Univ Illinois, Dept Med, Div Nephrol, Chicago, IL USA. Cleveland Clin Fdn, Dept Biostat & Epidemiol, Cleveland, OH 44195 USA. Univ Miami, Div Nephrol, Dept Med, Miami, FL 33152 USA. NIDDK, Div Kidney Urol & Hematol Dis, Bethesda, MD USA. RP Wright, JT (reprint author), Case Western Reserve Univ, Dept Med, Div Hypertens, 10900 Euclid Ave,Wood Bldg,Room W-165, Cleveland, OH 44106 USA. FU NCRR NIH HHS [2P20 RR11104, P20-RR11145, 5M01 RR-00071, M01 RR-00080, M01 RR00827]; PHS HHS [M01 00032] NR 21 TC 79 Z9 81 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD JUL 22 PY 2002 VL 162 IS 14 BP 1636 EP 1643 DI 10.1001/archinte.162.14.1636 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 573GW UT WOS:000176821800013 PM 12123409 ER PT J AU Kresina, TF Flexner, CW Sinclair, J Correia, MA Stapleton, JT Adeniyi-Jones, S Cargill, V Cheever, LW AF Kresina, TF Flexner, CW Sinclair, J Correia, MA Stapleton, JT Adeniyi-Jones, S Cargill, V Cheever, LW TI Alcohol use and HIV pharmacotherapy SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Review ID HUMAN-IMMUNODEFICIENCY-VIRUS; HEPATITIS-C VIRUS; ACTIVE ANTIRETROVIRAL THERAPY; LIVER FIBROSIS PROGRESSION; PRIMARY-CARE PHYSICIANS; SEXUAL RISK-TAKING; INFECTED PATIENTS; DRUG-USERS; PROTEASE INHIBITOR; SUBSTANCE USE AB Alcohol consumption by individuals infected with HIV is an important medical management issue with significant implications for the effectiveness of antiretroviral therapy as well as an important evolving field of HIV research. Alcohol consumption is a risk factor for poor medication adherence and can modify liver drug metabolism, both of which can lead to the emergence of drug-resistant virus. Research indicates that alcohol consumption greater than 50 g/day (four or five drinks) is a risk factor for liver disease progression among patients with HIV/HCV coinfection. In addition, alcohol-induced cirrhosis can result in changes in drug metabolism in the liver through compromised liver function. More research studies are needed to elucidate the biological and molecular basis of the clinical changes induced by alcohol consumption in HIV-infected individuals and on the relationship of these changes to the effectiveness of HIV pharmacotherapy. Specifically, research areas that are of particular importance are (1) determining alcohol consumption levels and patterns and its impact on antiretroviral medication adherence, efficacy, and physician prescribing practices; (2) identifying behavioral interventions to enhance adherence to HIV medications and reduce alcohol consumption; (3) clarifying the relationships and interactions among alcohol metabolism, HIV drug metabolism, and pharmacogenetics; (4) elucidating the extent of liver toxicity due to antiretroviral therapy and drug-drug interactions in individuals who consume alcohol; and (5) delineating the contribution of alcohol consumption to end-stage organ damage, particularly in HIV/HCV coinfection. C1 US Hlth Resources & Serv Adm, HIV Educ Branch, HIV AIDS Bur, Rockville, MD 20857 USA. NIDA, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Baltimore, MD 21205 USA. Vet Affairs Med Ctr, Hanover, NH 03755 USA. Dartmouth Coll Sch Med, Hanover, NH 03755 USA. Univ Calif San Francisco, Sch Med, San Francisco, CA 94143 USA. Univ Iowa, Iowa City, IA 52246 USA. Vet Affairs Med Ctr, Iowa City, IA 52246 USA. NIAID, NIH, Bethesda, MD 20892 USA. NIH, Off AIDS Res, Bethesda, MD 20892 USA. RP Cheever, LW (reprint author), US Hlth Resources & Serv Adm, HIV Educ Branch, HIV AIDS Bur, Parklawn Bldg,Room 7-29,5600 Fishers Lane, Rockville, MD 20857 USA. NR 114 TC 51 Z9 51 U1 2 U2 5 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD JUL 20 PY 2002 VL 18 IS 11 BP 757 EP 770 DI 10.1089/08892220260139495 PG 14 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 577AD UT WOS:000177038600003 PM 12167267 ER PT J AU Russo, P Malacarne, D Falugi, C Trombino, S O'Connor, PM AF Russo, P Malacarne, D Falugi, C Trombino, S O'Connor, PM TI RPR-115135, a farnesyltransferase inhibitor, increases 5-FU- cytotoxicity in ten human colon cancer cell lines: Role of p53 SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE farnesyltransferase; apoptosis; 5-FU; cell cycle; p53 ID FARNESYL TRANSFERASE INHIBITORS; SIGNAL-TRANSDUCTION PATHWAY; ANTICANCER DRUG SCREEN; COLORECTAL-CANCER; GERANYLGERANYLTRANSFERASE-I; PROTEIN TRANSFERASE; RAS MUTATION; APOPTOSIS; THERAPY; SENSITIVITY AB A new non peptidic farnesyltransferase inhibitor, RPR-115135, in combination with 5-FU was studied in 10 human colon cancer cell lines (HCT-116, RKO, DLD-1, Colo- 320, LoVo, SW-620, HT-29, HCT-15, Colo-205 and KM-12) carrying several mutations but well characterized for p53 and Ras status. We found that there was a slight tendency (not statistically significant) for the p53 inactivated cells to be less sensitive to S-FU after 6 days continuous treatment. Simultaneous administration of RPR- 115135 and 5-FU, at subtoxic concentrations, resulted in a synergistic enhancement of S-FU cytotoxicity in the p53 wildtype cells (HCT-116, RKO, DLD-1, Colo-320, LoVo). In the p53 mutated cells (SW-620, HT-29, HCT- IS, Colo-205, KM-12) the effect was very complicated. In HCT- IS the combination resulted in antagonism, in KM-12 in antagonism or in synergy (at different concentrations) and in SW-620, HT-29 and Colo-205 cells in synergy but only when S-FU was administered at high concentrations. Growth inhibition could be accounted for on the basis of a specific cell cycle arrest phenotype (G2-M arrest), as assayed by flow cytometry, only in the p53 functioning cell lines. The combination RPR-115135 + 5-FU increases apoptotic events only in these cell lines. In the mutated cell lines no major alterations on cell cycle arrest phenotype and no induction of apoptosis was observed. Although RPR- 115 135 can potentiate the effect of S-FU in cells in which p53 function is disrupted, these data suggest strongly that RPR- 115 135 significantly enhances the efficacy of S-FU only when p53 is functioning. (C) 2002 Wiley-Liss, Inc. C1 Natl Inst Canc Res, Expt Oncol Lab, Mol Pathol Sect, I-16132 Genoa, Italy. Univ Genoa, DIBISAA, Dept Expt Environm & Appl Biol, Genoa, Italy. NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Russo, P (reprint author), Natl Inst Canc Res, Expt Oncol Lab, Mol Pathol Sect, Largo Rosanna Benzi 10, I-16132 Genoa, Italy. NR 47 TC 36 Z9 36 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD JUL 20 PY 2002 VL 100 IS 3 BP 266 EP 275 DI 10.1002/ijc.10461 PG 10 WC Oncology SC Oncology GA 571DF UT WOS:000176699000003 PM 12115540 ER PT J AU Lorincz, AT Castle, PE Sherman, ME Scott, DR Glass, AG Wacholder, S Rush, BB Gravitt, PE Schussler, JE Schiffman, M AF Lorincz, AT Castle, PE Sherman, ME Scott, DR Glass, AG Wacholder, S Rush, BB Gravitt, PE Schussler, JE Schiffman, M TI Viral load of human papillomavirus and risk of CIN3 or cervical cancer SO LANCET LA English DT Article ID INTRAEPITHELIAL NEOPLASIA AB Carcinogenic human papillomaviruses (HPV) are thought to be necessary for development of cervical cancer. We assessed whether higher viral loads of such viruses predicted future risk of CIN3 or cancer (CIN3+) in a cohort of 20 810 women followed up for 10 years with cytological screening. We measured the viral load for 13 types of carcinogenic HPV (relative light units normalised to 1 pg/mL HPV 16 positive controls [RLU/PC]) using Hybrid Capture 2 testing of cervicovaginal lavages obtained at enrolment. Results were stratified into four groups (RLU/PC 1 to <10, 10 to <100, 100 to <1000, &GE;1000). Although presence of HPV strongly Increased risk of CIN3+, high viral load did not further predict risk of CIN3+. C1 Digene Corp, Gaithersburg, MD 20878 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Kaiser Permanente, Portland, OR USA. Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Baltimore, MD USA. Informat Management Serv Inc, Silver Spring, MD USA. RP Lorincz, AT (reprint author), Digene Corp, 1201 Clopper Rd, Gaithersburg, MD 20878 USA. NR 5 TC 148 Z9 177 U1 0 U2 5 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUL 20 PY 2002 VL 360 IS 9328 BP 228 EP 229 DI 10.1016/S0140-6736(02)09463-1 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 576DX UT WOS:000176988500016 PM 12133661 ER PT J AU Patera, AC Pesnicak, L Bertin, J Cohen, JI AF Patera, AC Pesnicak, L Bertin, J Cohen, JI TI Interleukin 17 modulates the immune response to vaccinia virus infection SO VIROLOGY LA English DT Article DE interleukin-17; NK cell cytotoxicity; herpesvirus; vaccinia virus infection ID KILLER-CELL-ACTIVITY; NATURAL-KILLER; HERPESVIRUS SAIMIRI; T-CELLS; GROWTH TRANSFORMATION; CYTOKINE RECEPTOR; NK CELL; IL-17; MICE; GENE AB Interleukin 17 (IL-17) is a newly identified cytokine that has a homolog in herpesvirus saimiri. We inserted murine IL-17 into vaccinia virus to study the role of IL-17 in viral infection. Vaccinia virus expressing IL-17 (vv-IL17) and its parental control virus (vv-pRB) grew to similar titers in vitro; however, vv-IL17 was more virulent in mice with a threefold lower LD50 than for vv-pRB, and mean time to death of 2.8 days versus 4.5 days. Mice infected with vv-IL17 had higher titers of virus in the ovaries (P < 0.02) and showed a decrease in NK cell cytotoxicity (P < 0.02) on day 3 after infection. No significant difference was found in CTL activity. In addition, a significant decrease in IgG2a (P < 0.01) and increases in IgG1, IgG3, and IgA (P < 0.03) vaccinia virus-specific antibody titers were observed in mice infected with vv-IL17 versus vv-pRB, suggesting a Th-2-like response to infection. These data indicate that IL-17 modulates the immune response during virus infection. C1 NIAID, Med Virol Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Cohen, JI (reprint author), NIAID, Med Virol Sect, Clin Invest Lab, NIH, Bldg 10,Room 11N228,10 Ctr Dr, Bethesda, MD 20892 USA. NR 40 TC 33 Z9 37 U1 1 U2 5 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD JUL 20 PY 2002 VL 299 IS 1 BP 56 EP 63 DI 10.1006/viro.2002.1400 PG 8 WC Virology SC Virology GA 585KW UT WOS:000177524800007 PM 12167341 ER PT J AU Hoshino, Y Jones, RW Kapikian, AZ AF Hoshino, Y Jones, RW Kapikian, AZ TI Characterization of neutralization specificities of outer capsid spike protein VP4 of selected murine, lapine, and human rotavirus strains SO VIROLOGY LA English DT Article ID INDUCED DIARRHEA; MONOCLONAL-ANTIBODIES; VACCINE DEVELOPMENT; SEROTYPE; PROTECTION; MICE; IDENTIFICATION; INFECTION; IMMUNITY; MODEL AB Neutralization specificities of outer capsid spike protein VP4 of murine rotavirus strains EW (P?[16],G3) and EHP (P?[20],G3) and lapine rotavirus strains Ala (P?[14],G3), C11 (P?[14],G3), and R2 (P?[14],G3) as well as human rotavirus strains PA169 (P?[14],G6) and HAL1166 (P?[14],G8) were determined by two-way cross-neutralization. This was done by generating and characterizing (i) three murine X human, three lapine X human, and two human X human single gene substitution reassortant rotaviruses, each of which bore identical human rotavirus DS-1 strain VP7 (G2), and (ii) guinea pig hyperimmune antiserum raised against each reassortant. Reference rotavirus strains employed in the study represented 10 established VP4 (P) serotypes, including 1A[8], 1B[4], 2A[6], 3[9], 4[10], 5A[2], 5B[2], 5B[3], 6[1], 7[5], 8[11], 9[7], and 10[16] as well as a P serotype unknown P[18]. Murine rotavirus strains EW and EB were demonstrated to share the same P serotype (P10[16]) distinct from (i) 9 established P serotypes, (ii) lapine and human rotavirus strains bearing the P[14] genotype, and (iii) an equine rotavirus strain bearing the P[18] genotype. Both lapine (Ala, C11, and R2) and human (PA169 and HAL1166) rotaviruses were shown to belong to the same VP4 serotype, which represented a distinct new P serotype (P11[14]). P serotype 13[20] was assigned to murine rotavirus EHP strain VP4, which was shown to be distinct from all the P serotypes/genotypes examined in the present study. C1 NIAID, ES, LID, NIH, Bethesda, MD 20892 USA. RP Hoshino, Y (reprint author), NIAID, ES, LID, NIH, Bldg 50,Room 6308,50 S Dr MSC 8026, Bethesda, MD 20892 USA. NR 47 TC 32 Z9 35 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD JUL 20 PY 2002 VL 299 IS 1 BP 64 EP 71 DI 10.1006/viro.2002.1474 PG 8 WC Virology SC Virology GA 585KW UT WOS:000177524800008 PM 12167342 ER PT J AU Mogayzel, PJ Esmail, M Wagner, TL Zeitlin, PL Ashlock, M AF Mogayzel, PJ Esmail, M Wagner, TL Zeitlin, PL Ashlock, M TI cis-Acting elements within CFTR 5 '-flanking DNA are not sufficient to decrease gene expression in response to phorbol ester SO BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION LA English DT Article DE cystic fibrosis; phorbol ester; CFTR expression ID TRANSMEMBRANE CONDUCTANCE REGULATOR; CYSTIC-FIBROSIS GENE; YEAST ARTIFICIAL CHROMOSOMES; I-HYPERSENSITIVE SITES; TRANSGENIC MICE; PROMOTER REGION; DOWN-REGULATION; MESSENGER-RNA; CELL LINE; CAMP AB The cystic fibrosis transmembrane conductance regulator gene (CFTR) is regulated in a tissue-specific and developmental fashion. Although it has been known for some time that phorbol esters decrease CFTR expression in cell lines that have high CFTR mRNA levels, the cis-acting elements that control this down-regulation remain ill-defined. The role of cis-acting elements within the CFTR minimal promoter in modulating responses to phorbol 12-myristate 13-acetate (PMA) and forskolin was assessed using luciferase reporter gene (luc)-containing plasmids transfected into Calu-3 and HT-29 cells. PMA treatment had no effect on luciferase activity in Calu-3 cells transiently transfected with plasmids containing luc driven by up to 2.3 kb of CFTR 5'-flanking DNA. PMA increased luciferase activity in transfected HT-29 cells. A more extensive region of DNA was evaluated using a yeast artificial chromosome (YAC) containing luc driven by similar to335 of CFTR 5'-flanking DNA (y5' luc) stably introduced into HT-29 cells. Clonal cell lines containing y5' luc were created and assessed for luciferase activity at baseline and in response to forskolin and PMA. There was a wide range of baseline luciferase activities among the clones (421038 units/mug protein) that was not entirely due to the number of luc copies present within the cells. Treatment with both PMA and forskolin led to increased luciferase activity in six randomly selected clonal cell lines. As expected. endogenous CFTR expression increased in response to forskolin and decreased in response to PMA. The-se studies demonstrate that lire-containing YAC vectors can be used to study CFTR expression in human cells. In addition, these data suggest that important regulatory elements responsible for decreased CFTR expression in response to PMA are not located upstream of CFTR in the similar to335 kb 5'-flanking sequence included in this YAC construct. (C) 2002 Elsevier Science B.V All rights reserved. C1 Johns Hopkins Med Inst, Eudowood Div Pediat Resp Sci, Baltimore, MD 21287 USA. NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. RP Mogayzel, PJ (reprint author), Johns Hopkins Med Inst, Eudowood Div Pediat Resp Sci, 600 N Wolfe St,Pk 316, Baltimore, MD 21287 USA. NR 50 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4781 J9 BBA-GENE STRUCT EXPR JI Biochim. Biophys. Acta-Gene Struct. Expression PD JUL 19 PY 2002 VL 1576 IS 3 BP 306 EP 315 AR PII S0167-4781(02)00395-0 DI 10.1016/S0167-4781(02)00395-0 PG 10 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 571KK UT WOS:000176716400008 PM 12084577 ER PT J AU Porter, AC Bymaster, FP DeLapp, NW Yamada, M Wess, J Hamilton, SE Nathanson, NM Felder, CC AF Porter, AC Bymaster, FP DeLapp, NW Yamada, M Wess, J Hamilton, SE Nathanson, NM Felder, CC TI M1 muscarinic receptor signaling in mouse hippocampus and cortex SO BRAIN RESEARCH LA English DT Article DE M1 muscarinic acetylcholine receptor; antibody capture GTP-binding; receptor reserve; Furchgott analysis; hippocampus; cortex ID ACETYLCHOLINE-RECEPTOR; MICE LACKING; CHOLINERGIC HYPOTHESIS; CORTICAL-NEURONS; SUBTYPE; IDENTIFICATION; LOCALIZATION; CELLS; BRAIN; GENE AB The five subtypes (M1-M5) of muscarinic acetylcholine receptors signal through Galphaq or Galphai/Galphao. M1, M3 and M5 receptors couple through Galphaq and function predominantly as postsynaptic receptors in the central nervous system. M1 and M3 receptors are localized to brain regions involved in cognition, such as hippocampus and cortex. but their relative contribution to function has been difficult to ascertain due to the lack of subtype specific ligands. A functional and genetic approach was used to identify the predominant muscarinic receptor subtype(s) mediating responses in mouse hippocampus and cortex, is well as the relative degree of spare muscarinic receptors in hippocampus. The nonselective muscarinic agonist oxotremorine-M stimulated Galphaq/11-specific GTP-gamma-S-35 binding in a concentration dependent manner with a Hill slope near unity in wild type mouse hippocampus and cortex. Muscarinic receptor stimulated Galphaq/11-specific GTP-gamma-S-35 binding was virtually abolished in both the hippocampus and cortex of M1 receptor knockout (KO) mice. In contrast, there was no loss of signaling in M3 receptor KO ruice in either brain region. Muscarinic receptor reserve in wildtype mouse hippocampus was measured by Furchgott analysis after partial receptor alkylation with propylbenzylcholine mustard. Occupation of just 15% of the MI receptors in mouse hippocampus was required for maximal efficacy of oxotrernorine-M -stimulated GTP- S bind ng indicating a substantial level of spare receptors. These findings support a role for the M1 receptor subtype as the primary Gaq/11-coupled muscarinic receptor in mouse hippocampus and cortex. (C) 2002 Elsevier Science B.V. All rights reserved. C1 Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs, Neurosci Res Div, Indianapolis, IN 46285 USA. NIDDK, Bioorgan Chem Lab, Bethesda, MD 20892 USA. Univ Washington, Sch Med, Dept Pharmacol, Seattle, WA 98195 USA. RP Porter, AC (reprint author), Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs, Neurosci Res Div, Indianapolis, IN 46285 USA. FU NINDS NIH HHS [NS26920] NR 27 TC 57 Z9 64 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD JUL 19 PY 2002 VL 944 IS 1-2 BP 82 EP 89 AR PII S0006-8993(02)02721-X DI 10.1016/S0006-8993(02)02721-X PG 8 WC Neurosciences SC Neurosciences & Neurology GA 580FH UT WOS:000177224100009 PM 12106668 ER PT J AU Spiegel, S Milstien, S AF Spiegel, S Milstien, S TI Sphingosine 1-phosphate, a key cell signaling molecule. SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Review ID PROTEIN-COUPLED RECEPTOR; SACCHAROMYCES-CEREVISIAE; LYSOPHOSPHATIDIC ACID; INTRACELLULAR SPHINGOSINE-1-PHOSPHATE; FUNCTIONAL-CHARACTERIZATION; CALCIUM MOBILIZATION; 2ND MESSENGER; PDGF-B; KINASE; APOPTOSIS C1 Virginia Commonwealth Univ, Dept Biochem, Richmond, VA 23298 USA. NIH, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. RP Spiegel, S (reprint author), Virginia Commonwealth Univ, Dept Biochem, Med Coll Virginia Campus, Richmond, VA 23298 USA. FU NCI NIH HHS [CA61774]; NIGMS NIH HHS [GM43880] NR 73 TC 388 Z9 405 U1 1 U2 11 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 19 PY 2002 VL 277 IS 29 BP 25851 EP 25854 DI 10.1074/jbc.R200007200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 574UV UT WOS:000176908700004 PM 12011102 ER PT J AU Kolter, T Proia, RL Sandhoff, K AF Kolter, T Proia, RL Sandhoff, K TI Combinatorial ganglioside biosynthesis. SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Review ID GALACTOSE-CERAMIDE GALACTOSYLTRANSFERASE; RAT-LIVER GOLGI; COMPLEX GANGLIOSIDES; NERVOUS-SYSTEM; GLYCOSPHINGOLIPID BIOSYNTHESIS; GLYCOLIPID BIOSYNTHESIS; ENDOPLASMIC-RETICULUM; EXPRESSION CLONING; RECEPTOR FUNCTION; MEMBRANE-PROTEIN C1 Kekule Inst Organ Chem & Biochem, D-53121 Bonn, Germany. NIDDK, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. RP Sandhoff, K (reprint author), Kekule Inst Organ Chem & Biochem, Gerhard Domagk Str 1, D-53121 Bonn, Germany. RI Proia, Richard/A-7908-2012 NR 73 TC 190 Z9 193 U1 0 U2 14 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 19 PY 2002 VL 277 IS 29 BP 25859 EP 25862 DI 10.1074/jbc.R200001200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 574UV UT WOS:000176908700006 PM 12011101 ER PT J AU Chipuk, JE Stewart, LV Ranieri, A Song, K Danielpour, D AF Chipuk, JE Stewart, LV Ranieri, A Song, K Danielpour, D TI Identification and characterization of a novel rat ov-serpin family member, trespin SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PLASMINOGEN-ACTIVATOR INHIBITOR; GROWTH-FACTOR-BETA; PROSTATE EPITHELIAL-CELLS; GRANZYME-B INHIBITOR; PROTEINASE-INHIBITORS; CRYSTAL-STRUCTURE; TGF-BETA; TRANSFORMING GROWTH-FACTOR-BETA-1; ALPHA-1-PROTEINASE INHIBITOR; PROTEASE INHIBITOR-10 AB Serpins are responsible for regulating a variety of proteolytic processes through a unique irreversible suicide substrate mechanism. To discover novel genes regulated by transforming growth factor-beta1 (TGF-beta1), we performed differential display reverse transcriptase-PCR analysis of NRP-152 rat prostatic epithelial cells and cloned a novel rat serpin that is transcriptionally down-regulated by TGF-beta and hence named trespin (TGF-beta-repressible serine proteinase inhibitor (trespin). Trespin is a 397-amino acid member of the ov-serpin clade with a calculated molecular mass of 45.2 kDa and 72% amino acid sequence homology to human bomapin; however, trespin exhibits different tissue expression, cellular localization, and proteinase specificity compared with bomapin. Trespin mRNA is expressed in many tissues, including brain, heart, kidney, liver, lung, prostate, skin, spleen, and stomach. FILAG-trespin expressed in HEK293 cells is localized predominantly in the cytoplasm and is not constitutively secreted. The presence of an arginine at the P1 position of trespin's reactive site loop suggests that trespin inhibits trypsin-like proteinases. Accordingly, in vitro transcribed and translated trespin forms detergent-stable and thermostable complexes with plasmin and elastase but not subtilisin A, trypsin, chymotrypsin, thrombin, or papain. Trespin interacts with plasmin at a near 1:1 stoichiometry, and immunopurified mammal-expressed trespin inhibits plasmin in a dose-dependent manner. These data suggest that trespin is a novel and functional member of the rat ov-serpin family. C1 Ireland Canc Ctr, Res Labs, Cleveland, OH 44106 USA. Case Western Reserve Univ Hosp, Dept Pharmacol, Cleveland, OH 44106 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Danielpour, D (reprint author), Ireland Canc Ctr, Res Labs, Samuel Gerber Bldg,Suite 200,Lab 3,11001 Cedar Rd, Cleveland, OH 44106 USA. OI Chipuk, Jerry Edward/0000-0002-1337-842X FU NCI NIH HHS [1R01 CA3069-01, P30CA43703, R01 CA083069] NR 69 TC 6 Z9 8 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 19 PY 2002 VL 277 IS 29 BP 26412 EP 26421 DI 10.1074/jbc.M20124420 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 574UV UT WOS:000176908700078 PM 11986314 ER PT J AU Lener, D Budihas, SR Le Grice, SFJ AF Lener, D Budihas, SR Le Grice, SFJ TI Mutating conserved residues in the ribonuclease H domain of Ty3 reverse transcriptase affects specialized cleavage events SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DNA STRAND TRANSFER; RNA SECONDARY STRUCTURE; YEAST RETROTRANSPOSON TY1; PRIMER TRANSFER-RNA; ESCHERICHIA-COLI; ACTIVE-SITE; NUCLEOCAPSID PROTEIN; CRYSTAL-STRUCTURE; POLYPURINE TRACT; POLYMERASE AB The reverse transcriptase-associated ribonuclease H (RT/RNase H) domains from the gypsy group of retrotransposons, of which Ty3 is a member, share considerable sequence homology with their retroviral counterparts. However, the gypsy elements have a conserved tyrosine (position 459 in Ty3 RT) instead of the conserved histidine in the catalytic center of retroviral RTs such as at position 539 of HIV-1. In addition, the gypsy group shows conservation of histidine adjacent to the third of the metal-chelating carboxylate residues, which is Asp-426 of Ty3 RT. The role of these and additional catalytic residues was assessed with purified recombinant enzymes and through the ability of Ty3 mutants to support transposition in Saccaromyces cerevisiae. Although all mutations had minimal impact on DNA polymerase function, amidation of Asp-358, Glu-401, and Asp-426 eliminated Me2+ and Mn2+-dependent RNase H function. Replacing His-427 and Tyr-459 with Ala and Asp-469 with Asn resulted in reduced RNase H activity in the presence of Mg2+, whereas in the presence of Mn2+ these mutants displayed a lack of turnover. Despite this, mutations at all positions were lethal for transposition. To reconcile these apparently contradictory findings, the efficiency of specialized RNase H-mediated events was examined for each enzyme. Mutants retaining RNase H activity on a heteropolymeric RNA-DNA hybrid failed to support DNA strand transfer and release of the (+) strand polypurine tract primer from (+) RNA, suggesting that interrupting one or both of these events might account for the transposition defect. C1 NCI, Reverse Transcriptase Biochem Sect, Resistance Mechanisms Lab, HIV Drug Resistance Program,NIH, Frederick, MD 21702 USA. RP Le Grice, SFJ (reprint author), NCI, Reverse Transcriptase Biochem Sect, Resistance Mechanisms Lab, HIV Drug Resistance Program,NIH, 1050 Boyles St,POB B, Frederick, MD 21702 USA. NR 60 TC 18 Z9 18 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 19 PY 2002 VL 277 IS 29 BP 26486 EP 26495 DI 10.1074/jbc.M200496200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 574UV UT WOS:000176908700087 PM 11994277 ER PT J AU Kirby, TW Mueller, GA DeRose, EF Lebetkin, MS Meiss, G Pingoud, A London, RE AF Kirby, TW Mueller, GA DeRose, EF Lebetkin, MS Meiss, G Pingoud, A London, RE TI The nuclease A inhibitor represents a new variation of the rare PR-1 fold SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE nuclease A inhibitor; NuiA; nuclease A; NucA; Anabaena ID PATHOGENESIS-RELATED PROTEIN; 3-DIMENSIONAL SOLUTION STRUCTURE; NMR STRUCTURE CALCULATION; CHEMICAL-SHIFT INDEX; RESONANCE SPECTROSCOPY; DISTANCE RESTRAINTS; SECONDARY STRUCTURE; BACKBONE DYNAMICS; ENDONUCLEASE-G; CLASSIFICATION AB Nuclease A (NucA) from Anabaena sp. is a non-specific endonuclease able to degrade single and double-stranded DNA and RNA. The endonucleolytic activity is inhibited by the nuclease A inhibitor (NuiA), which binds to NucA with 1:1 stoichiometry and picomolar affinity. In order to better understand the mechanism of inhibition, the solution structure of NuiA was determined by NMR methods. The fold of NuiA is an alpha-beta-alpha sandwich but standard database searches by DALI and TOP revealed no structural homologies. A visual inspection of alpha-beta-alpha folds in the CATH database revealed similarities to the PR-1-like fold (SCOP nomenclature). The similarities include the ordering of secondary structural elements, a single helix on one face of the alpha-beta-alpha sandwich, and three helices on the other face. However, a major difference is in the IV helix, which in the PR-1 fold is short and perpendicular to the I and III helices, but in NuiA is long and parallel to the I and III helices. Additionally, a strand insertion in the beta-sheet makes the NuiA beta-sheet completely antiparallel in organization. The fast time-scale motions of NuiA, characterized by enhanced flexibility of the extended loop between helices III and IV, also show similarities to P14a, which is a PR-1 fold. We propose that the purpose of the PR-1 fold is to form a stable scaffold to present this extended structure for biological interactions with other proteins. This hypothesis is supported by data that show that when NuiA is bound to NucA significant changes in chemical shift occur in the extended loop between helices III and IV. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 NIEHS, Res Triangle Pk, NC 27709 USA. Univ Giessen, Inst Biochem, D-35392 Giessen, Germany. RP London, RE (reprint author), NIEHS, POB 12233,MD MR-01, Res Triangle Pk, NC 27709 USA. NR 61 TC 14 Z9 14 U1 1 U2 3 PU ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD JUL 19 PY 2002 VL 320 IS 4 BP 771 EP 782 DI 10.1061/S0022-2836(02)00460-6 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 581DU UT WOS:000177276500008 PM 12095254 ER PT J AU Moody, TW Czerwinski, G Tarasova, NI Michejda, CJ AF Moody, TW Czerwinski, G Tarasova, NI Michejda, CJ TI VIP-ellipticine derivatives inhibit the growth of breast cancer cells SO LIFE SCIENCES LA English DT Article DE VIP-ellipticine derivatives; VIP agonists; VPAC(1) receptor; breast cancer ID FUNCTIONAL EXPRESSION; RECEPTOR; PEPTIDE; POLYPEPTIDE; ANTAGONIST; TUMORS; PACAP; LUNG AB The effects of vasoactive intestinal peptide (VIP)-ellipticine (E) derivatives were investigated on breast cancer cells. VIP-ALALA-E and VIP-LALA-E inhibited I-125-VIP binding to MCF-7 cells with an IC50 values of 1 and 0.2 muM respectively. VIP-ALALA-E and VIP-LALA-E caused elevation of cAMP in MCF-7 cells with ED50 values of 1 and 0.1 muM. VIP-LALA-E caused increased c-fos mRNA in MCF-7 cells. Radiolabeled VIP-LALA-E was internalized at 37 C and delivered the cytotoxic E into MCF-7 cells. VIP-LALA-E inhibited the clonal growth of MCF-7 cells, decreased cell viability based on trypan blue exclusion and reduced S-35-methionine uptake. These results indicate that VIP-E derivatives function as breast cancer VPAC(1) receptor agonists which inhibit MCF-7 cellular viability. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NCI, Cell & Canc Biol Branch, Ctr Canc Res, Rockville, MD 20850 USA. NCI, Mol Aspects Drug Design Sect, Macromol Struct Lab, Adv BioSci Labs, Frederick, MD 21702 USA. RP Moody, TW (reprint author), NCI, Cell & Canc Biol Branch, Ctr Canc Res, Rockville, MD 20850 USA. NR 23 TC 29 Z9 31 U1 1 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0024-3205 J9 LIFE SCI JI Life Sci. PD JUL 19 PY 2002 VL 71 IS 9 BP 1005 EP 1014 AR PII S0024-3205(02)01741-1 DI 10.1016/S0024-3205(02)01741-1 PG 10 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 583BX UT WOS:000177386600003 PM 12088760 ER PT J AU O'Farrell, C Hernandez, DG Evey, C Singleton, AB Cookson, MR AF O'Farrell, C Hernandez, DG Evey, C Singleton, AB Cookson, MR TI Normal localization of Delta F323-Y328 mutant torsinA in transfected human cells SO NEUROSCIENCE LETTERS LA English DT Article DE dystonia; torsinA; subcellular localization; endoplasmic reticulum ID DYSTONIA; ONSET; MUTATION; FORMS; GENE AB Two mutations in torsinA have been identified to date, both of which are associated with an autosomal dominant form of early onset-dystonia. It has been reported previously that expression of the more common mutation, a deletion of one of a pair of glutamates (DeltaE302/303) produces intracellular, endoplasmic reticulum-derived inclusions in cultured cells. In this study we have replicated these previous results and have additionally looked at the localization of the more recently described DeltaF323-Y328 mutation. We show that the localization of this latter mutation is similar to wild type torsinA and unlike the DeltaE302/303 mutation. This data suggests that the formation of intracellular inclusions is specific to DeltaE302/303 and not a property shared by DeltaF323-Y328. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Mayo Clin, Dept Neurosci, Jacksonville, FL 32224 USA. NIA, NIH, Neurogenet Lab, Bethesda, MD 20892 USA. RP Cookson, MR (reprint author), Mayo Clin, Dept Neurosci, 4500 San Pablo Rd, Jacksonville, FL 32224 USA. RI Singleton, Andrew/C-3010-2009 FU NINDS NIH HHS [P01-NS40256, R01-NS41816-01] NR 10 TC 20 Z9 20 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD JUL 19 PY 2002 VL 327 IS 2 BP 75 EP 78 AR PII S0304-3940(02)00400-7 DI 10.1016/S0304-3940(02)00400-7 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 582UP UT WOS:000177369800001 PM 12098639 ER PT J AU Tachibana, T Noguchi, K Ruda, MA AF Tachibana, T Noguchi, K Ruda, MA TI Analysis of gene expression following spinal cord injury in rat using complementary DNA microarray SO NEUROSCIENCE LETTERS LA English DT Article DE microarray; spinal cord injury; reverse transcription-polymerase chain reaction; heat shock protein; tissue inhibitor of metalloproteinase; fatty acid-binding protein; lecithin; cholesterol acyltransferase; plasma membrane Ca2+-ATPase ID CENTRAL-NERVOUS-SYSTEM; DIFFERENTIAL EXPRESSION; BINDING PROTEIN; POLYPEPTIDE 7B2; FAMILY; CELLS AB To identify genes that were altered by spinal cord injury (SCI), we used complementary DNA microarray consisting 1176 rat genes. Rats were subjected to contusive injury of the thoracic spinal cord. Sham animals received only a laminectomy. Twenty-four hours later, spinal cord was dissected out, a P-32 labeled probe was prepared and hybridized to the microarray. We identified three genes that showed a greater than 2-fold increase in SCI tissue, heat shock 27-kDa protein, tissue inhibitor of metalloproteinase-1 and epidermal fatty acid-binding protein. Seven genes, lecithin:cholesterol acyltransferase, dipeptidyl aminopeptidase related protein, phospholipase C delta 4, plasma membrane Ca2+ ATPase isoform 2, G-protein G(O) alpha subunit, GABA transporter 3, and neuroendrocrine protein 7132 were downregulated greater than 50% in SCI tissue. Changes in expression of these genes were confirmed by reverse transcription-polymerase chain reaction. These genes may play a role in the response to tissue damage or repair following SCI. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 NIDCR, Cellular Neurosci Sect, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. Hyogo Med Univ, Dept Anat & Neurosci, Nishinomiya, Hyogo 6638501, Japan. RP Tachibana, T (reprint author), NIDCR, Cellular Neurosci Sect, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. NR 20 TC 53 Z9 55 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD JUL 19 PY 2002 VL 327 IS 2 BP 133 EP 137 AR PII S0304-3940(02)00375-0 DI 10.1016/S0304-3940(02)00375-0 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 582UP UT WOS:000177369800015 PM 12098653 ER PT J AU Schmidt, LA Fox, NA Rubin, KH Hu, S Hamer, DH AF Schmidt, LA Fox, NA Rubin, KH Hu, S Hamer, DH TI Molecular genetics of shyness and aggression in preschoolers SO PERSONALITY AND INDIVIDUAL DIFFERENCES LA English DT Article DE shyness; aggression; molecular genetics; DRD4; 5-HTT; 5-HT2C; preschoolers ID SEROTONIN TRANSPORTER GENE; EXON-III POLYMORPHISM; HUMAN PERSONALITY-TRAIT; RECEPTOR D4 DRD4; DEFICIT HYPERACTIVITY DISORDER; NOVELTY-SEEKING; DOPAMINE-D4 RECEPTOR; BEHAVIORAL-INHIBITION; REPEAT POLYMORPHISM; REGULATORY REGION AB Associations between three candidate gene polymorphisms [dopamine D4 receptor (DRD4), serotonin transporter (5-HTT), and serotonin 2C receptor (5-HT2C)] with shy and aggression-related behaviors derived from maternal report and peer play at age four were examined. We noted a significant association of the DRD4 receptor gene with maternal report of problems with aggression at age four. Children with long versus short repeat alleles of the DRD4 gene were reported by their mothers to have significantly more problems with aggression at age four. There were no significant associations of the DRD4 gene with observed behavioral measures of aggression at age four. There were, in addition, no significant associations of either of the serotonin genes with any of the maternal report and observed behavioral measures. The present study extends earlier findings of adults to the preschool years and appears to be the first large scale investigation to examine the molecular genetics of preschoolers' temperament using behavioral and maternal report measures in normal childhood development. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 McMaster Univ, Dept Psychol, Hamilton, ON L8S 4K1, Canada. Univ Maryland, Inst Child Study, College Pk, MD 20742 USA. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Schmidt, LA (reprint author), McMaster Univ, Dept Psychol, Hamilton, ON L8S 4K1, Canada. NR 48 TC 68 Z9 71 U1 1 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0191-8869 J9 PERS INDIV DIFFER JI Pers. Individ. Differ. PD JUL 19 PY 2002 VL 33 IS 2 BP 227 EP 238 AR PII S0191-8869(01)00147-7 DI 10.1016/S0191-8869(01)00147-7 PG 12 WC Psychology, Social SC Psychology GA 577DW UT WOS:000177047100003 ER PT J AU Long, CA Hoffman, SL AF Long, CA Hoffman, SL TI Parasitology: Malaria - from infants to genomics to vaccines SO SCIENCE LA English DT Editorial Material C1 NIH, Malaria Vaccine Dev Unit, Rockville, MD 20852 USA. Celera Genom, Rockville, MD 20850 USA. RP Long, CA (reprint author), NIH, Malaria Vaccine Dev Unit, Rockville, MD 20852 USA. NR 2 TC 14 Z9 15 U1 0 U2 0 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD JUL 19 PY 2002 VL 297 IS 5580 BP 345 EP 347 DI 10.1126/science.1074484 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574LV UT WOS:000176892600033 PM 12130768 ER PT J AU Hardy, J Selkoe, DJ AF Hardy, J Selkoe, DJ TI Medicine - The amyloid hypothesis of Alzheimer's disease: Progress and problems on the road to therapeutics SO SCIENCE LA English DT Review ID BETA-PROTEIN-PRECURSOR; HEREDITARY CEREBRAL-HEMORRHAGE; TRANSGENIC MOUSE MODEL; CENTRAL-NERVOUS-SYSTEM; SPASTIC PARAPARESIS; IN-VIVO; FRONTOTEMPORAL DEMENTIA; PRESENILIN-1 MUTATIONS; PLAQUE-FORMATION; GENETIC-LINKAGE C1 Harvard Univ, Brigham & Womens Hosp, Sch Med, Ctr Neurol Dis, Boston, MA 02115 USA. NIA, Neurogenet Lab, Bethesda, MD 20892 USA. RP Selkoe, DJ (reprint author), Harvard Univ, Brigham & Womens Hosp, Sch Med, Ctr Neurol Dis, Boston, MA 02115 USA. RI Hardy, John/C-2451-2009 NR 91 TC 6664 Z9 6991 U1 297 U2 1883 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD JUL 19 PY 2002 VL 297 IS 5580 BP 353 EP 356 DI 10.1126/science.1072994 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574LV UT WOS:000176892600038 PM 12130773 ER PT J AU Hariri, AR Mattay, VS Tessitore, A Kolachana, B Fera, F Goldman, D Egan, MF Weinberger, DR AF Hariri, AR Mattay, VS Tessitore, A Kolachana, B Fera, F Goldman, D Egan, MF Weinberger, DR TI Serotonin transporter genetic variation and the response of the human amygdala SO SCIENCE LA English DT Article ID ANXIETY-RELATED TRAITS; PERSONALITY-TRAITS; POLYMORPHISM; NEUROTICISM; ASSOCIATION; DISORDERS; GENOTYPE; EXPRESSION; CORTEX; BRAIN AB A functional polymorphism in the promoter region of the human serotonin transporter gene (SLC6A4) has been associated with several dimensions of neuroticism and psychopathology, especially anxiety traits, but the predictive value of this genotype against these complex behaviors has been inconsistent. Serotonin [5-hydroxytryptamine, (5-HT)] function influences normal fear as well as pathological anxiety, behaviors critically dependent on the amygdala in animal models and in clinical studies. We now report that individuals with one or two copies of the short allele of the serotonin transporter (5-HTT) promoter polymorphism, which has been associated with reduced 5-HTT expression and function and increased fear and anxiety-related behaviors, exhibit greater amygdala neuronal activity, as assessed by BOLD functional magnetic resonance imaging, in response to fearful stimuli compared with individuals homozygous for the long allele. These results demonstrate genetically driven variation in the response of brain regions underlying human emotional behavior and suggest that differential excitability of the amygdala to emotional stimuli may contribute to the increased fear and anxiety typically associated with the short SLC6A4 allele. C1 NIMH, Clin Brain Disorders Branch, Intramural Res Program, NIH, Bethesda, MD 20892 USA. NIAAA, Neurogenet Lab, Intramural Res Program, NIH, Bethesda, MD 20892 USA. RP Weinberger, DR (reprint author), NIMH, Clin Brain Disorders Branch, Intramural Res Program, NIH, 10 Ctr Dr,Room 4S235, Bethesda, MD 20892 USA. RI Hariri, Ahmad/D-5761-2011; Goldman, David/F-9772-2010 OI Goldman, David/0000-0002-1724-5405 NR 37 TC 1415 Z9 1445 U1 18 U2 145 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD JUL 19 PY 2002 VL 297 IS 5580 BP 400 EP 403 DI 10.1126/science.1071829 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574LV UT WOS:000176892600051 PM 12130784 ER PT J AU Pais, GCG Zhang, XC Marchand, C Neamati, N Cowansage, K Svarovskaia, ES Pathak, VK Tang, Y Nicklaus, M Pommier, Y Burke, TR AF Pais, GCG Zhang, XC Marchand, C Neamati, N Cowansage, K Svarovskaia, ES Pathak, VK Tang, Y Nicklaus, M Pommier, Y Burke, TR TI Structure activity of 3-aryl-1,3-diketo-containing compounds as HIV-1 integrase inhibitors SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID DRUG DESIGN; STRAND TRANSFER; TARGET; CELLS; REPLICATION; COMPLEXES; ACYLATION; INFECTION; DYNAMICS; ACIDS AB The 4-aryl-2-hydroxy-4-oxo-2-butenoic acids and their isosteric tetrazoles are among an emerging class of aryl beta-diketo (ADK)-based agents which exhibit potent inhibition of HIV-1 integrase (IN)-catalyzed strand transfer (ST) processes, while having much reduced potencies against X-processing (3'-P) reactions. In the current study, L-708,906 (10e) and 5CITEP (13b), which are two examples of ADK inhibitors that have been reported by Merck and Shionogi pharmaceutical companies, served as model ADK leads. Structural variations to both the "left" and "right" sides of these molecules were made in order to examine effects on HIV-1 integrase inhibitory potencies. It was found that a variety of groups could be introduced onto the left side aryl ring with maintenance of good ST inhibitory potency. However, introduction of carboxylic acid-containing substituents onto the left side aryl ring enhanced T-P inhibitory potency and reduced selectivity toward ST reactions. Although both L-708,906 and 5CITEP show potent inhibition of IN in biochemical assays, there is a disparity of antiviral activity in cellular assays using HIV-1-infected cells. Neither 5CITEP nor any other of the indolyl-containing inhibitors exhibit significant antiviral effects in cellular systems. Alternatively, consistent with literature reports, L-708,906 does provide antiviral protection at low micromolar concentrations. Interestingly, several analogues of L-708,906 with varied substituents on the left side aryl ring, while having good inhibitory potencies against IN in extracellular assays, are not antiviral in whole-cell systems. C1 NCI, Ctr Canc Res, Med Chem Lab, NIH, Frederick, MD 21702 USA. NCI, Ctr Canc Res, Mol Pharmacol Lab, NIH, Frederick, MD 21702 USA. NCI, Ctr Canc Res, HIV Drug Resistance Program, NIH, Frederick, MD 21702 USA. RP Burke, TR (reprint author), NCI, Ctr Canc Res, Med Chem Lab, NIH, Frederick, MD 21702 USA. RI Burke, Terrence/N-2601-2014; Marchand, Christophe/D-8559-2016; OI Nicklaus, Marc/0000-0002-4775-7030 NR 44 TC 156 Z9 163 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD JUL 18 PY 2002 VL 45 IS 15 BP 3184 EP 3194 DI 10.1021/jm020037p PG 11 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 573TB UT WOS:000176846000003 PM 12109903 ER PT J AU Evan-Ram, S AF Evan-Ram, S TI Names: a historical or political perspective? SO NATURE LA English DT Letter C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD 20892 USA. RP Evan-Ram, S (reprint author), Natl Inst Dent & Craniofacial Res, NIH, 30 Convent Dr, Bethesda, MD 20892 USA. OI Even-Ram, Sharona/0000-0002-5540-3822 NR 1 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 18 PY 2002 VL 418 IS 6895 BP 273 EP 273 DI 10.1038/418273a PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574BF UT WOS:000176868000014 ER PT J AU Wootton, JC Feng, XR Ferdig, MT Cooper, RA Mu, JB Baruch, DI Magill, AJ Su, XZ AF Wootton, JC Feng, XR Ferdig, MT Cooper, RA Mu, JB Baruch, DI Magill, AJ Su, XZ TI Genetic diversity and chloroquine selective sweeps in Plasmodium falciparum SO NATURE LA English DT Article ID HUMAN MALARIA; PARASITE POPULATIONS; RECENT ORIGIN; RESISTANCE; PROTEIN; POLYMORPHISMS; PFCRT AB Widespread use of antimalarial agents can profoundly influence the evolution of the human malaria parasite Plasmodium falciparum. Recent selective sweeps for drug-resistant genotypes may have restricted the genetic diversity of this parasite, resembling effects attributed in current debates(1-4) to a historic population bottleneck. Chloroquine-resistant (CQR) parasites were initially reported about 45 years ago from two foci in southeast Asia and South America(5), but the number of CQR founder mutations and the impact of chlorquine on parasite genomes worldwide have been difficult to evaluate. Using 342 highly polymorphic microsatellite markers from a genetic map(6), here we show that the level of genetic diversity varies substantially among different regions of the parasite genome, revealing extensive linkage disequilibrium surrounding the key CQR gene pfcrt(7) and at least four CQR founder events. This disequilibrium and its decay rate in the pfcrt-flanking region are consistent with strong directional selective sweeps occurring over only similar to20-80 sexual generations, especially a single resistant pfcrt haplotype spreading to very high frequencies throughout most of Asia and Africa. The presence of linkage disequilibrium provides a basis for mapping genes under drug selection in P. falciparum. C1 NIAID, Lab Malaria & Vector Res, NIH, Bethesda, MD 20892 USA. NIH, Computat Biol Branch, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. Univ Notre Dame, Dept Biol Sci, Notre Dame, IN 46556 USA. Walter Reed Army Med Ctr, Walter Reed Army Inst Res, Div Communicable Dis & Immunol, Washington, DC 20307 USA. RP Su, XZ (reprint author), NIAID, Lab Malaria & Vector Res, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. RI feng, xiaorong/G-4811-2010; Ferdig, Michael/C-6627-2016; OI feng, xiaorong/0000-0001-8410-3020; Su, Xinzhuan/0000-0003-3246-3248 NR 20 TC 460 Z9 468 U1 2 U2 41 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 18 PY 2002 VL 418 IS 6895 BP 320 EP 323 DI 10.1038/nature00813 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574BF UT WOS:000176868000041 PM 12124623 ER PT J AU Mu, JB Duan, JH Makova, KD Joy, DA Huynh, CQ Branch, OH Li, WH Su, XZ AF Mu, JB Duan, JH Makova, KD Joy, DA Huynh, CQ Branch, OH Li, WH Su, XZ TI Chromosome-wide SNPs reveal an ancient origin for Plasmodium falciparum SO NATURE LA English DT Article ID SEQUENCE; PROTEIN; POLYMORPHISMS; POPULATIONS; COMPLEXITY; REGIONS; BOTTLENECK; SELECTION; EVOLUTION; MALARIAS AB The Malaria's Eve hypothesis, proposing a severe recent population bottleneck (about 3,000-5,000 years ago) of the human malaria parasite Plasmodium falciparum, has prompted a debate about the origin and evolution of the parasite(1-6). The hypothesis implies that the parasite population is relatively homogeneous, favouring malaria control measures. Other studies, however, suggested an ancient origin and large effective population size(5,7-10). To test the hypothesis, we analysed single nucleotide polymorphisms (SNPs) from 204 genes on chromosome 3 of P. falciparum. We have identified 403 polymorphic sites, including 238 SNPs and 165 microsatellites, from five parasite clones, establishing chromosome-wide haplotypes and a dense map with one polymorphic marker per similar to2.3 kilobases. On the basis of synonymous SNPs and non-coding SNPs, we estimate the time to the most recent common ancestor to be similar to100,000-180,000 years, significantly older than the proposed bottleneck. Our estimated divergence time coincides approximately with the start of human population expansion(11), and is consistent with a genetically complex organism able to evade host immunity and other antimalarial efforts. C1 NIAID, Lab Malaria & Vector Res, NIH, Bethesda, MD 20892 USA. Univ Chicago, Dept Ecol & Evolut, Chicago, IL 60637 USA. NIH, Informat Engn Branch, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. NIH, Computat Biol Branch, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. RP Su, XZ (reprint author), NIAID, Lab Malaria & Vector Res, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Su, Xinzhuan/0000-0003-3246-3248; Li, Wen-Hsiung/0000-0003-1086-1456 NR 29 TC 105 Z9 110 U1 0 U2 4 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 18 PY 2002 VL 418 IS 6895 BP 323 EP 326 DI 10.1038/nature00836 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 574BF UT WOS:000176868000042 PM 12124624 ER PT J AU Koechlin, E Danek, A Brunod, Y Grafman, J AF Koechlin, E Danek, A Brunod, Y Grafman, J TI Medial prefrontal and subcortical mechanisms underlying the acquisition of motor and cognitive action sequences in humans SO NEURON LA English DT Article ID POSITRON-EMISSION-TOMOGRAPHY; ANTERIOR CINGULATE CORTEX; VENTRAL STRIATUM; BASAL GANGLIA; PARKINSONS-DISEASE; NEURONAL-ACTIVITY; WORKING-MEMORY; REWARD; MONKEY; ORGANIZATION AB The anterior media[ prefrontal cortex (AMPC) in humans is involved in affect and in regulating goal-directed behaviors. The precise function of the AMPC, however, is poorly understood. Using magnetic resonance imaging, we found that bilateral regions in the AMPC were selectively recruited to compute the reliability of subjects' expectations that developed when subjects were learning sequences of cognitive tasks. In contrast, regions similarly recruited in learning sequences of motor acts were found in the ventral striatum. Our results show that beyond the execution of motor acts, the AMPC is selectively engaged in computing the relevance of cognitive goals that subjects intend to achieve. This indicates that the fronto-striatal circuit, including the ventral striatum and AMPC, subserves hierarchically distinct evaluative processes mediating the human ability to build behavioral plans, ranging from motor to cognitive action plans. C1 Univ Paris 06, INSERM, U483, F-75005 Paris, France. NINDS, Cognit Neurosci Sect, Bethesda, MD 20892 USA. RP Koechlin, E (reprint author), Univ Paris 06, INSERM, U483, 9 Quai St Bernard, F-75005 Paris, France. EM koechlin@ccr.jussieu.fr RI Danek, Adrian/G-7339-2011; Koechlin, Etienne/E-5061-2016; OI Danek, Adrian/0000-0001-8857-5383; Grafman, Jordan H./0000-0001-8645-4457 NR 50 TC 55 Z9 55 U1 1 U2 4 PU CELL PRESS PI CAMBRIDGE PA 600 TECHNOLOGY SQUARE, 5TH FLOOR, CAMBRIDGE, MA 02139 USA SN 0896-6273 J9 NEURON JI Neuron PD JUL 18 PY 2002 VL 35 IS 2 BP 371 EP 381 DI 10.1016/S0896-6273(02)00742-0 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 576YF UT WOS:000177034200016 PM 12160754 ER PT J AU Liang, TJ Ghany, M AF Liang, TJ Ghany, M TI Hepatitis B e antigen - The dangerous endgame of hepatitis B SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material ID HEPATOCELLULAR-CARCINOMA; VIRUS; VACCINATION; CHILDREN; OUTCOMES; TAIWAN C1 NIDDKD, Bethesda, MD 20892 USA. RP Liang, TJ (reprint author), NIDDKD, Bethesda, MD 20892 USA. NR 16 TC 13 Z9 16 U1 1 U2 1 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUL 18 PY 2002 VL 347 IS 3 BP 208 EP 210 DI 10.1056/NEJMe020060 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 574BR UT WOS:000176869000009 PM 12124411 ER PT J AU Emerling, BM Bonifas, J Kratz, CP Donovan, S Taylor, BR Green, ED Le Beau, MM Shannon, KM AF Emerling, BM Bonifas, J Kratz, CP Donovan, S Taylor, BR Green, ED Le Beau, MM Shannon, KM TI MLL5, a homolog of Drosophila trithorax located within a segment of chromosome band 7q22 implicated in myeloid leukemia SO ONCOGENE LA English DT Article DE monosomy 7; deletion 7q; myeloid leukemia; trithorax genes ID BONE-MARROW CELLS; MYELOGENOUS LEUKEMIA; MUTANT MICE; GENE; FUSION; REGION; NEUROFIBROMATOSIS; T(7-11)(P15-P15); TRANSLOCATIONS; LEUKEMOGENESIS AB Proteins encoded by Polycomb and Trithorax-group (Pc-G and Trx-G) genes regulate developmental fates by maintaining or repressing HOX gene expression, respectively. In a search for candidate myeloid leukemia tumor suppressor genes from a similar to2.5 Mb commonly-deleted segment within chromosome band 7q22, we identified a novel human Trithorax (Trx) family member named MLL5. Trx-G genes encode proteins that modulate transcriptional programs through protein-protein interactions that are mediated by PHD and SET domains, and by binding to DNA via A-T hooks and methyltransferase homology motifs. MLL5 is a homolog of the Drosophila gene CG9007; it encodes a 6.5 kb mRNA that is expressed widely. MLL5 includes a SET domain and a single PHD finger, but lacks A-T hooks and methyltransferase homology domains that are found in MLL. The leukemia cell line RCV-ACV-A carries a heterozygous missense mutation within the PHD domain; however, no mutations within the MLL5 coding region were detected in primary leukemias. MLL5 is a novel mammalian Trx-G gene that might modulate transcription by protein association. C1 Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94143 USA. Univ Calif San Francisco, Ctr Comprehens Canc, San Francisco, CA 94143 USA. NHGRI, Genome Technol Branch, Bethesda, MD 20892 USA. Univ Chicago, Hematol Oncol Sect, Dept Med, Chicago, IL 60637 USA. Univ Chicago, Ctr Canc Res, Chicago, IL 60637 USA. RP Shannon, KM (reprint author), Univ Calif San Francisco, Dept Pediat, 513 Parnassus Ave,HSE 302, San Francisco, CA 94143 USA. FU NCI NIH HHS [CA 40046] NR 35 TC 66 Z9 69 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUL 18 PY 2002 VL 21 IS 31 BP 4849 EP 4854 DI 10.1038/sj.onc.1205615 PG 6 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 571KJ UT WOS:000176716300015 PM 12101424 ER PT J AU Zhou, L Nepote, V Rowley, DL Levacher, B Zvara, A Santha, M Mi, QS Simonneau, M Donovan, DM AF Zhou, L Nepote, V Rowley, DL Levacher, B Zvara, A Santha, M Mi, QS Simonneau, M Donovan, DM TI Murine peripherin gene sequences direct Cre recombinase expression to peripheral neurons in transgenic mice SO FEBS LETTERS LA English DT Article DE ROSA26; dorsal root ganglion; sensory neuron; Cre; loxP; peripherin ID INTERMEDIATE FILAMENT PROTEIN; SITE-SPECIFIC RECOMBINATION; MOUSE EMBRYOS; BACTERIOPHAGE-P1; KNOCKOUT; BRAIN AB Spatially and temporally regulated somatic mutations can be achieved by using the Cre/loxP recombination system of bacteriophage PI. To develop a cell type-specific system of gene targeting in the peripheral nervous system, we generated the transgenic mouse lines expressing Cre recombinase under the control of the mouse peripherin gene promoter. The activity of the Cre recombinase during embryonic development was examined by mating the peripherin-Cre transgenic mice to the knock-in Cre-mediated recombination reporter strain, R26R. Analysis of F1 embryos from this cross showed specific excision of loxP-flanked sequences in the dorsal root ganglia, trigeminal ganglia, and olfactory epithelium, in a pattern very similar to the expression of the endogenous mouse peripherin gene, and the previously reported peripherin-lacZ transgenic mice. Thus, the peripherin-Cre mouse described here will provide a valuable tool for Cre-loxP-mediated conditional expression in the peripheral nervous system. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies. C1 NIA, Gerontol Res Ctr, Transgen & Knockout Facil Sect, IRP,NIH, Baltimore, MD 21224 USA. Hop Robert Debre, INSERM E9935, F-75019 Paris, France. NIA, Gerontol Res Ctr, Immunol Lab, IRP,NIH, Baltimore, MD 21224 USA. RP Donovan, DM (reprint author), NIA, Gerontol Res Ctr, Transgen & Knockout Facil Sect, IRP,NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Santha, Miklos/F-4772-2013 NR 23 TC 18 Z9 18 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD JUL 17 PY 2002 VL 523 IS 1-3 BP 68 EP 72 AR PII S0014-5793(02)02936-8 DI 10.1016/S0014-5793(02)02936-8 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 580DK UT WOS:000177219300014 PM 12123806 ER PT J AU Lenders, JWM Pacak, K Eisenhofer, G AF Lenders, JWM Pacak, K Eisenhofer, G TI Imaging vs biochemical testing for pheochromocytoma - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter ID ENDOCRINE NEOPLASIA TYPE-2; LINDAU-DISEASE C1 St Radboud Univ, Med Ctr, Nijmegen, Netherlands. NIH, Bethesda, MD 20892 USA. RP Lenders, JWM (reprint author), St Radboud Univ, Med Ctr, Nijmegen, Netherlands. RI Lenders, J.W.M./L-4487-2015 NR 4 TC 1 Z9 1 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD JUL 17 PY 2002 VL 288 IS 3 BP 315 EP 315 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 573AK UT WOS:000176807000019 ER PT J AU Rossouw, JE Anderson, GL Prentice, RL LaCroix, AZ Kooperberg, C Stefanick, ML Jackson, RD Beresford, SAA Howard, BV Johnson, KC Kotchen, JM Ockene, J AF Rossouw, JE Anderson, GL Prentice, RL LaCroix, AZ Kooperberg, C Stefanick, ML Jackson, RD Beresford, SAA Howard, BV Johnson, KC Kotchen, JM Ockene, J TI Risks and benefits of estrogen plus progestin in healthy postmenopausal women - Principal results from the Women's Health Initiative randomized controlled trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HORMONE REPLACEMENT THERAPY; BREAST-CANCER RISK; CORONARY-ARTERY ATHEROSCLEROSIS; CARDIOVASCULAR-DISEASE; SECONDARY PREVENTION; HEART-DISEASE; METAANALYSIS; EVENTS; ASSOCIATION; FRACTURES AB Context Despite decades of accumulated observational evidence, the balance of risks and benefits for hormone use in healthy postmenopausal women remains uncertain. Objective To assess the major health benefits and risks of the most commonly used combined hormone preparation in the United States. Design Estrogen plus progestin component of the Women's Health Initiative, a randomized controlled primary prevention trial (planned duration, 8.5 years) in which 16608 postmenopausal women aged 50-79 years with an intact uterus at baseline were recruited by 40 US clinical centers in 1993-1998. Interventions Participants received conjugated equine estrogens, 0.625 mg/d, plus medroxylprogesterone acetate, 2.5 mg/d, in 1 tablet (n = 8506) or placebo (n = 8102). Main Outcomes Measures The primary outcome was coronary heart disease (CHD) (nonfatal myocardial infarction and CHD death), with invasive breast cancer as the primary adverse outcome. A global index summarizing the balance of risks and benefits included the 2 primary outcomes plus stroke, pulmonary embolism (PE), endometrial cancer, colorectal cancer, hip fracture, and death due to other causes. Results On May 31, 2002, after a mean of 5.2 years of follow-up, the data and safety monitoring board recommended stopping the trial of estrogen plus progestin vs placebo because the test statistic for invasive breast cancer exceeded the stopping boundary for this adverse effect and the global index statistic supported risks exceeding benefits. This report includes data on the major clinical outcomes through April 30, 2002. Estimated hazard ratios (HRs) (nominal 95% confidence intervals [Cis]) were as follows: CHD, 1.29 (1.02-1.63) with 286 cases; breast cancer, 1.26 (1.00-1.59) with 290 cases; stroke, 1.41 (1.07-1.85) with 212 cases; PE, 2.13 (1.39-3.25) with 101 cases; colorectal cancer, 0.63 (0.43-0.92) with 112 cases; endometrial cancer, 0.83 (0.47-1.47) with 47 cases; hip fracture, 0.66 (0.45-0.98) with 106 cases; and death due to other causes, 0.92 (0.74-1.14) with 331 cases, Corresponding HRs (nominal 95% Cis) for composite outcomes were 1.22 (1.09-1.36) for total cardiovascular disease (arterial and venous disease), 1,03 (0.90-1.17) for total cancer, 0.76 (0.69-0.85) for combined fractures, 0.98 (0.82-1.18) for total mortality, and 1.15 (1.03-1.28) for the global index. Absolute excess risks per 10000 person-years attributable to estrogen plus progestin were 7 more CHD events, 8 more strokes, 8 more PEs, and 8 more invasive breast cancers, while absolute risk reductions per 10000 1 person-years were 6 fewer colorectal cancers and 5 fewer hip fractures. The absolute excess risk of events included in the global index was 19 per 10000 person-years. Conclusions Overall health risks exceeded benefits from use of combined estrogen plus progestin for an average 5.2-year follow-up among healthy postmenopausal US women. All-cause mortality was not affected during the trial. The risk-benefit profile found in this trial is not consistent with the requirements for a viable intervention for primary prevention of chronic diseases, and the results indicate that this regimen should not be initiated or continued for primary prevention of CHD. C1 NHLBI, Div Womens Hlth Initiat, Bethesda, MD 20817 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. Stanford Univ, Ctr Clin, Stanford, CA 94305 USA. Ohio State Univ, Ctr Clin, Columbus, OH 43210 USA. MedStar Res Inst, Washington, DC USA. Univ Tennessee, Memphis, TN USA. Med Coll Wisconsin, Milwaukee, WI 53226 USA. Univ Massachusetts, Sch Med, Worcester, MA USA. RP Rossouw, JE (reprint author), NHLBI, Div Womens Hlth Initiat, 6705 Rockledge Dr,1 Rockledge Ctr,Suite 300,MS-79, Bethesda, MD 20817 USA. EM rossouw@nih.gov NR 43 TC 8344 Z9 8678 U1 78 U2 733 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60654-0946 USA SN 0098-7484 EI 1538-3598 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD JUL 17 PY 2002 VL 288 IS 3 BP 321 EP 333 PG 13 WC Medicine, General & Internal SC General & Internal Medicine GA 573AK UT WOS:000176807000024 ER PT J AU Lacey, JV Mink, PJ Lubin, JH Sherman, ME Troisi, R Hartge, P Schatzkin, A Schairer, C AF Lacey, JV Mink, PJ Lubin, JH Sherman, ME Troisi, R Hartge, P Schatzkin, A Schairer, C TI Menopausal hormone replacement therapy and risk of ovarian cancer SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID NONCONTRACEPTIVE ESTROGEN USE; STATES CASE-CONTROL; BREAST-CANCER; UNITED-STATES; COLLABORATIVE ANALYSIS; EPITHELIAL-CELLS; RECEPTOR-ALPHA; MESSENGER-RNA; FOLLOW-UP; WOMEN AB Context The association between menopausal hormone replacement therapy and ovarian cancer is unclear. Objective To determine whether hormone replacement therapy using estrogen only, estrogen-progestin only, or both estrogen only and estrogen-progestin increases ovarian cancer risk. Design A 1979-1998 cohort study of former participants in the Breast Cancer Detection Demonstration Project, a nationwide breast cancer screening program. Setting Twenty-nine US clinical centers. Participants A total of 44241 postmenopausal women (mean age at start of follow-up, 56.6 years). Main Outcome Measure Incident ovarian cancer. Results We identified 329 women who developed ovarian cancer during follow-up. In time-dependent analyses adjusted for age, menopause type, and oral contraceptive use, ever use of estrogen only was significantly associated with ovarian cancer (rate ratio [RR], 1.6; 95% confidence interval [CI], 1.2-2.0). Increasing duration of estrogen-only use was significantly associated with ovarian cancer: RRS for 10 to 19 years and 20 or more years were 1.8 (95% Cl, 1.1-3.0) and 3.2 (95% Cl, 1.7-5.7), respectively (P value for trend <.001), and we observed a 7% (95% Cl, 2%-13%) increase in RR per year of use. We observed significantly elevated RRs with increasing duration of estrogen-only use across all strata of other ovarian cancer risk factors, including women with hysterectomy. The RR for estrogen-progestin use after prior estrogen-only use was 1.5 (95% Cl, 0.91-2.4), but the RR for estrogen-progestin-only use was 1.1 (95% Cl, 0.64-1.7). The RRs for less than 2 years and 2 or more years of estrogen-progestin-only use were 1.6 (95% Cl, 0.78-3.3) and 0.80 (95% Cl, 0.35-1.8), respectively, and there was no evidence of a duration response (P value for trend =30). Conclusion Women who used estrogen-only replacement therapy, particularly for 10 or more years, were at significantly increased risk of ovarian cancer in this study. Women who used short-term estrogen-progestin-only replacement therapy were not at increased risk, but risk associated with short-term and longer-term estrogen-progestin replacement therapy warrants further investigation. C1 NCI, Div Canc Epidemiol & Genet, Rockville, MD 20852 USA. RP Lacey, JV (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,MSC 7234, Rockville, MD 20852 USA. NR 48 TC 342 Z9 371 U1 2 U2 11 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD JUL 17 PY 2002 VL 288 IS 3 BP 334 EP 341 DI 10.1001/jama.288.3.334 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 573AK UT WOS:000176807000025 PM 12117398 ER PT J AU Johnson, PC Wheat, LJ Cloud, GA Goldman, M Lancaster, D Bamberger, DM Powderly, WG Hafner, R Kauffman, CA Dismukes, WE AF Johnson, PC Wheat, LJ Cloud, GA Goldman, M Lancaster, D Bamberger, DM Powderly, WG Hafner, R Kauffman, CA Dismukes, WE CA US Natl Inst Allergy & Infect Dis TI Safety and efficacy of liposomal amphotericin B compared with conventional amphotericin B for induction therapy of histoplasmosis in patients with AIDS SO ANNALS OF INTERNAL MEDICINE LA English DT Article ID ACQUIRED-IMMUNODEFICIENCY-SYNDROME; CENTRAL-NERVOUS-SYSTEM; CAPSULATUM ANTIGEN; FLUCONAZOLE; DIAGNOSIS AB Background: In patients with moderate to severe histoplasmosis associated with AIDS, the preferred treatment has been the deoxycholate formulation of amphotericin B. However, serious side effects are associated with use of amphotericin B. Objective: To compare amphotericin B with liposomal amphotericin B for induction therapy of moderate to severe disseminated histoplasmosis in patients with AIDS. Design: Randomized, double-blind, multicenter clinical trial. Setting: 21 sites of the U.S. National Institute of Allergy and Infectious Diseases Mycoses Study Group. Patients: 81 patients with AIDS and moderate to severe disseminated histoplasmosis. Measurements: Clinical success, conversion of baseline blood cultures to negative, and acute toxicities that necessitated discontinuation of treatment. Results: Clinical success was achieved in 14 of 22 patients (64%) treated with amphotericin B compared with 45 of 51 patients (88%) receiving liposomal amphotericin B (difference, 24 percentage points [95% Cl, 1 to 52 percentage points]). Culture conversion rates were similar. Three patients treated with amphotericin B and one treated with liposomal amphotericin 8 died during induction (P = 0.04). Infusion-related side effects were greater with amphotericin B (63%) than with liposomal amphotericin B (25%) (P = 0.002). Nephrotoxicity occurred in 37% of patients treated with amphotericin B and 9% of patients treated with liposomal amphotericin B (P = 0.003). Conclusion: Liposomal amphotericin B seems to be a less toxic alternative to amphotericin B and is associated with improved survival. C1 Univ Texas, Sch Med, Div Gen Med, Houston, TX 77030 USA. Indiana Univ, Sch Med, Indianapolis, IN 46204 USA. Richard L Roudebush Vet Affairs Med Ctr, Indianapolis, IN 46202 USA. Univ Alabama, Birmingham, AL USA. Methodist Hosp, Memphis, TN USA. Univ Missouri, Kansas City, MO 64110 USA. Washington Univ, Sch Med, St Louis, MO USA. NIAID, Mycoses Study Grp, NIH, Bethesda, MD 20892 USA. Vet Affairs Med Ctr, Ann Arbor, MI USA. Univ Michigan, Sch Med, Ann Arbor, MI 48109 USA. RP Johnson, PC (reprint author), Univ Texas, Sch Med, Div Gen Med, 6431 Fannin,MSB 1-122, Houston, TX 77030 USA. FU NCRR NIH HHS [MO1-RR02558]; PHS HHS [N01-A1-15802] NR 17 TC 122 Z9 131 U1 0 U2 1 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 J9 ANN INTERN MED JI Ann. Intern. Med. PD JUL 16 PY 2002 VL 137 IS 2 BP 105 EP 109 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 575VN UT WOS:000176969300004 PM 12118965 ER PT J AU Tarasov, SG Nelson, C Cholody, WM Hariprakasha, H Kosakowska-Cholody, T Casas-Finet, JR Michejda, CJ AF Tarasov, SG Nelson, C Cholody, WM Hariprakasha, H Kosakowska-Cholody, T Casas-Finet, JR Michejda, CJ TI Physicochemical study of DNA binding properties of the naphthylimido-imidazoacridone WMC79 and related compounds. SO BIOCHEMISTRY LA English DT Meeting Abstract C1 NCI, Mol Aspects Drug Design Sect, Struct Biophys Lab, Frederick, MD 21702 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 16 PY 2002 VL 41 IS 28 MA 160 BP 8996 EP 8997 PG 2 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 572LW UT WOS:000176777000179 ER PT J AU O'Donnell, CJ Chazaro, I Wilson, PWF Fox, C Hannan, MT Kiel, DP Cupples, LA AF O'Donnell, CJ Chazaro, I Wilson, PWF Fox, C Hannan, MT Kiel, DP Cupples, LA TI Evidence for heritability of abdominal aortic calcific deposits in the Framingham Heart Study SO CIRCULATION LA English DT Article DE atherosclerosis; calcification; abdominal aorta; genetics; heritability ID CORONARY-ARTERY CALCIFICATION; RISK-FACTORS; VASCULAR CALCIFICATION; DISEASE; POLYMORPHISM; AGGREGATION; FAMILIES; LESIONS; GENE AB Background-Atherosclerosis is a systemic disease that underlies clinical cardiovascular disease. The radiographic finding of abdominal aortic calcific deposits is an indicator of the presence of aortic atherosclerosis and an independent predictor of cardiovascular disease events. Little is known about the heritability of aortic calcification. Methods and Results-Original Framingham Heart Study cohort participants (2151) in 1109 extended pedigrees had a lateral lumbar radiograph. The presence and severity of abdominal aortic calcific (AAC) deposits at the levels of the first through fourth lumbar vertebrae was graded by a previously validated rating scale. Correlation coefficients were calculated in pairs of siblings, parent-offspring, and spouses. Age-, sex-, and multivariable-adjusted correlation coefficients for AAC were 0.52 for parent-offspring pairs and 0.20 for sibling pairs. In contrast, the multivariable-adjusted correlation for AAC in spouse pairs was -0.02. Using variance component methods implemented in SOLAR, the estimated heritability for age-, sex-, and multivariable-adjusted AAC was 0.49 (P<0.001). Thirty-one percent of the overall variance in AAC deposits was due to measured covariates, and 49% to heritable factors. Conclusions-In our large, population-based sample, heritable factors play a role in the presence and extent of abdominal aortic calcification. Thus, a substantial proportion of the variation in AAC is due to additive effects of genes, which have yet to be characterized. Measures of aortic atherosclerosis may provide heritable quantitative phenotypes for the genetic dissection of the complex condition of atherosclerosis in human populations. C1 Natl Heart Lung & Blood Inst, Framingham Heart Study, Framingham, MA 01702 USA. Massachusetts Gen Hosp, Dept Med, Div Cardiol, Boston, MA USA. Hebrew Rehabil Ctr Aged, Res & Training Inst, Boston, MA USA. Harvard Univ, Sch Med, Div Aging, Boston, MA 02115 USA. Boston Univ, Sch Publ Hlth, Dept Epidemiol & Biostat, Boston, MA USA. NHLBI, NIH, Bethesda, MD 20892 USA. RP O'Donnell, CJ (reprint author), Natl Heart Lung & Blood Inst, Framingham Heart Study, 73 Mt Wayte Ave, Framingham, MA 01702 USA. OI Cupples, L. Adrienne/0000-0003-0273-7965; Kiel, Douglas/0000-0001-8474-0310 FU NHLBI NIH HHS [N01-HC-38038]; NIAMS NIH HHS [AR/AG 41398] NR 28 TC 52 Z9 53 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUL 16 PY 2002 VL 106 IS 3 BP 337 EP 341 DI 10.1161/01.CIR.0000022663.26468.5B PG 5 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 575JZ UT WOS:000176944300014 PM 12119250 ER PT J AU Engel, LS Taioli, E Pfeiffer, R Garcia-Closas, M Marcus, PM Lan, Q Boffetta, P Vineis, P Autrup, H Bell, DA Branch, RA Brockmoller, J Daly, AK Heckbert, SR Kalina, I Kang, DH Katoh, T Lafuente, A Lin, HJ Romkes, M Taylor, JA Rothman, N AF Engel, LS Taioli, E Pfeiffer, R Garcia-Closas, M Marcus, PM Lan, Q Boffetta, P Vineis, P Autrup, H Bell, DA Branch, RA Brockmoller, J Daly, AK Heckbert, SR Kalina, I Kang, DH Katoh, T Lafuente, A Lin, HJ Romkes, M Taylor, JA Rothman, N TI Pooled analysis and meta-analysis of glutathione S-transferase M1 and bladder cancer: A HuGE review SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Review DE bladder neoplasms; epidemiology; genetics; glutathione transferase ID CHLORINATION BY-PRODUCTS; LOWER URINARY-TRACT; MICROSOMAL EPOXIDE HYDROLASE; GENE-ENVIRONMENT INTERACTION; OCCUPATIONAL RISK-FACTORS; SQUAMOUS-CELL CARCINOMA; REDUCTION PLANT WORKERS; SMOKING-RELATED CANCERS; GSTM1 NULL GENOTYPE; CIGARETTE-SMOKING AB Smoking is a known risk factor for bladder cancer. The product of the GSTM1 gene, glutathione S-transferase M1 (GSTM1), is involved in the detoxification of polycyclic aromatic hydrocarbons found in tobacco smoke; a homozygous deletion of this gene in approximately 50% of Caucasians and Asians results in a lack of GSTM1 enzyme activity. Most studies examining the relation between bladder cancer and GSTM1 have reported an increased risk associated with a lack of GSTM1 activity. The authors performed meta- and pooled analyses of published and unpublished, case-control, genotype-based studies that examined this association (17 studies, 2,149 cases, 3,646 controls) and excluded studies conducted in populations with a high prevalence of exposure to known bladder cancer risk factors other than tobacco smoke. Using random effects models in the meta-analysis, the authors obtained a summary odds ratio of 1.44 (95% confidence interval (CI): 1.23, 1.68) for GSTM1 null status with all studies included. Results from studies with at least 100 cases and 100 controls produced a summary odds ratio of 1.42 (95% CI: 1.26, 1.60). Pooled analyses using original data sets from 10 studies (1,496 cases and 1,444 controls) and adjusting for age, sex, and race produced similar results. There was no evidence of multiplicative interaction between the GSTM1 null genotype and ever smoking in relation to bladder cancer, although there was a suggestion of additive interaction (additive interaction = 0.45, 95% CI: -0.03, 0.93). These results indicate that, among populations studied to date, GSTM1 null status is associated with a modest increase in the risk of bladder cancer. C1 NCI, Occupat Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ Milan, IRCCS, Milan, Italy. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. Int Agcy Res Canc, F-69372 Lyon, France. Univ Turin, Turin, Italy. Univ Aarhus, DK-8000 Aarhus C, Denmark. Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. Univ Pittsburgh, Pittsburgh, PA USA. Univ Goettingen, Dept Clin Pharmacol, Gottingen, Germany. Univ Newcastle Upon Tyne, Newcastle Upon Tyne, Tyne & Wear, England. Univ Washington, Seattle, WA 98195 USA. Safarik Univ, Kosice, Slovakia. Seoul Natl Univ, Seoul, South Korea. Miyazaki Med Coll, Miyazaki 88916, Japan. Univ Barcelona, Barcelona, Spain. Harbor UCLA Med Ctr, Torrance, CA 90509 USA. RP Engel, LS (reprint author), NCI, Occupat Epidemiol Branch, Div Canc Epidemiol & Genet, 6120 Execut Blvd,Room 8113, Bethesda, MD 20892 USA. RI Pfeiffer, Ruth /F-4748-2011; Daly, Ann/H-3144-2011; Kang, Dae Hee/E-8631-2012; Garcia-Closas, Montserrat /F-3871-2015; OI Daly, Ann/0000-0002-7321-0629; Garcia-Closas, Montserrat /0000-0003-1033-2650; taylor, jack/0000-0001-5303-6398 NR 150 TC 154 Z9 161 U1 3 U2 9 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUL 15 PY 2002 VL 156 IS 2 BP 95 EP 109 DI 10.1093/aje/kwf018 PG 15 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 574KN UT WOS:000176888600001 PM 12117698 ER PT J AU Harris, TB AF Harris, TB TI Invited commentary: Body composition in studies of aging: New opportunities to better understand health risks associated with weight SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE aging; body composition; cachexia; disability evaluation; methods; obesity ID HIP FRACTURE; COMPUTED-TOMOGRAPHY; MUSCLE MASS; WOMEN; MEN; SARCOPENIA; DISABILITY; EPIDEMIOLOGY; RATIOS; MODELS AB The expected increase in the proportion of older persons over the next century underscores the need to identify modifiable risk factors for disease and disability in this population. One such risk factor is weight, which plays a role in many of the diseases common in old age and contributes to risk of disability and death. However, there is confusion and controversy regarding health risks associated with weight in old age. The emergence of new technologies to assess body composition should allow opportunities to better understand health risks associated with weight in old age, as suggested by the new report in this issue of the Journal. While application of these technologies to population studies will still require careful attention to methodological caveats important in studies of weight, the ability to separately examine lean mass, bone, and fat should shed light on the underlying biologic processes pertinent to risk. C1 NIA, Geriatr Epidemiol Sect, Lab Epidemiol Demog & Biometry, NIH, Bethesda, MD 20892 USA. RP Harris, TB (reprint author), NIA, Geriatr Epidemiol Sect, Lab Epidemiol Demog & Biometry, NIH, 7201 Wisconsin Ave,3C-309, Bethesda, MD 20892 USA. NR 26 TC 26 Z9 27 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUL 15 PY 2002 VL 156 IS 2 BP 122 EP 124 DI 10.1093/aje/kwf024 PG 3 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 574KN UT WOS:000176888600003 PM 12117701 ER PT J AU Ng, D Hadley, DW Tifft, CJ Biesecker, LG AF Ng, D Hadley, DW Tifft, CJ Biesecker, LG TI Genetic heterogeneity of syndromic X-linked recessive microphthalmia-anophthalmia: Is Lenz microphthalmia a single disorder? SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE microphthalmia; X-linked; syndromic; linkage analysis ID ANOMALIES AB Nonsyndromic congenital microphthalmia or anophthalmia is a heterogeneous malformation with autosomal dominant, autosomal recessive, and X-linked modes of inheritance. Lenz microphthalmia syndrome comprises microphthalmia with mental retardation, malformed ears, skeletal anomalies, and is inherited in an X-linked recessive pattern. Prior studies have shown linkage of both isolated (or nonsyndromic) anophthalmos (ANOP1, [MINI 3015901) and Lenz syndrome [MIM 3098001 to Xq27-q28. Nonsyndromic colobomatous microphthalmia [MIM 3003451 has been linked to Xp11.4-Xq11.1. We describe a five-generation African-American family with microphthalmia or anophthalmia, mental retardation, and urogenital anomalies, in an X-linked recessive inheritance pattern, consistent with Lenz syndrome. Initial linkage analysis with microsatellite markers excluded the region in Xq27-q28 previously reported as a candidate region for ANOP1 [MIM 301590]. An X-chromosome scan revealed linkage to a 10-cM region between markers DXS228 and DXS992 in Xp11.4-p21.2. Multipoint analysis gave a maximum LOD score of 2.46 at marker DXS993. These data show that X-linked recessive syndromic microphthalmia exhibits genetic heterogeneity. In addition, it suggests that Lenz microphthalmia syndrome, previously thought to be a single disorder, may represent an amalgam of two distinct disorders. Published (C) 2002 Wiley-Liss, Inc. C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. NIDDK, Genet Dev & Dis Branch, NIH, Bethesda, MD USA. NHGRI, Genet Dis Res Branch, NIH, Bethesda, MD USA. RP Ng, D (reprint author), NHGRI, Med Genet Branch, NIH, Bldg 49,Room 4C72, Bethesda, MD 20892 USA. NR 19 TC 16 Z9 16 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD JUL 15 PY 2002 VL 110 IS 4 BP 308 EP 314 DI 10.1002/ajmg.10484 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 567PX UT WOS:000176494200002 PM 12116202 ER PT J AU Kumaki, F Takeda, K Yu, ZX Moss, J Ferrans, VJ AF Kumaki, F Takeda, K Yu, ZX Moss, J Ferrans, VJ TI Expression of human telomerase reverse transcriptase in lymphangioleiomyomatosis SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article DE lymphangioleiomyomatosis; lung; telomerase; smooth muscle cells ID CANCER-CELLS; PULMONARY LYMPHANGIOLEIOMYOMATOSIS; RNA COMPONENT; C-MYC; ACTIVATION; ASSOCIATION; FAMILY; HTERT AB Telomerase synthesizes nucleotide hexameric repeats (telomeres) at the ends of chromosomes, replacing base sequences that are lost from these sites during each mitotic cycle and protecting these ends against the action of exonucleases and ligases. Therefore, telomerase is essential for maintaining cellular replication. To evaluate the role of telomerase in the proliferation of abnormal smooth muscle cells (lymphangioleiomyomatosis [LAM] cells) in LAM, we performed immunostaining and in situ hybridization studies to identify telomerase protein and messenger RNA (mRNA), respectively, in pulmonary (n = 18) and extrapulmonary (n = 4) lesions from 22 women with LAM (14 untreated and 8 treated with progesterone or tamoxifen). Immunoreactivity and hybridization signals for telomerase were observed in 5 to 20% of LAM cells, mostly of the spindle-shaped type, in 21 of the 22 patients, and were less intense in the treated group. Other types of cells were unreactive in both groups. Telomerase colocalized in the same cells with a-smooth muscle actin, but only rarely with HMB-45 antibody (a marker for epithelioid LAM cells); colocalization with proliferating cell nuclear antigen was incomplete. The telomerase-positive LAM cells may constitute the sources of renewal of LAM cells. Modulation of telomerase may be involved in the control of LAM cell proliferation. C1 NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. RP Yu, ZX (reprint author), NHLBI, Pathol Sect, NIH, Bldg 10 2N240,10 Ctr Dr M5C-1S18, Bethesda, MD 20892 USA. NR 23 TC 8 Z9 10 U1 0 U2 1 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1073-449X J9 AM J RESP CRIT CARE JI Am. J. Respir. Crit. Care Med. PD JUL 15 PY 2002 VL 166 IS 2 BP 187 EP 191 DI 10.1164/rccm.2105045 PG 5 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 571XX UT WOS:000176745400013 PM 12119231 ER PT J AU Crystal, RG Bitterman, PB Mossman, B Schwarz, MI Sheppard, D Almasy, L Chapman, HA Friedman, SL King, TE Leinwand, LA Liotta, L Martin, GR Schwartz, DA Schultz, GS Wagner, CR Musson, RA AF Crystal, RG Bitterman, PB Mossman, B Schwarz, MI Sheppard, D Almasy, L Chapman, HA Friedman, SL King, TE Leinwand, LA Liotta, L Martin, GR Schwartz, DA Schultz, GS Wagner, CR Musson, RA TI Future research directions in idiopathic pulmonary fibrosis - Summary of a National Heart, Lung, and Blood Institute working group SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article DE lung diseases; interstitial; pulmonary fibrosis; National Institutes of Health (United States) ID GROWTH-FACTOR-BETA; OBLITERANS-ORGANIZING PNEUMONIA; LASER CAPTURE MICRODISSECTION; CONNECTIVE-TISSUE; GENE-EXPRESSION; IN-VIVO; FIBROBLAST PROLIFERATION; PROGNOSTIC-SIGNIFICANCE; INTERSTITIAL PNEUMONIA; SYSTEMIC-SCLEROSIS AB Idiopathic pulmonary fibrosis (IPF) is an insidious inflammatory fibroproliferative disease whose cause and course before diagnosis are unknown, and for which existing treatments are of limited benefit. The National Heart, Lung, and Blood Institute convened a working group to develop specific recommendations for future IPF research. Inflammatory and immune processes are involved in IPF pathogenesis, and current therapeutic strategies are aimed at suppressing the inflammation. Recent data suggest that the molecular processes underlying the fibrogenesis may provide new opportunities for therapeutic intervention. Specific areas of future research recommended by the working group include studies to elucidate the etiology of IPF, to develop novel diagnostic techniques and molecular diagnostics, to establish a program for identification of molecular targets for IPF treatment and identification and generation of agonists or antagonists that inhibit fibrogenesis, to foster investigations that couple the use of new technologies (e.g., laser capture microdissection, microarrays, and mass spectroscopic analysis of proteins) with data from the human genome project, to establish a national consortium of Clinical Centers of Excellence to conduct coordinated clinical and laboratory studies of well-characterized patients and patient-derived materials, and to stimulate research to develop animal models of persistent and progressive pulmonary fibrosis for evaluation of new intervention approaches. C1 Cornell Univ, Weill Med Coll, Div Pulm & Crit Care Med, New York, NY USA. Univ Minnesota, Sch Med, Dept Med, Minneapolis, MN 55455 USA. Univ Vermont, Dept Pathol, Burlington, VT 05405 USA. Univ Colorado, Hlth Sci Ctr, Denver, CO USA. Univ Calif San Francisco, Dept Med, San Francisco, CA USA. SW Fdn Biomed Res, Dept Genet, San Antonio, TX USA. Univ Calif San Francisco, Div Pulm & Crit Care, San Francisco, CA 94143 USA. Mt Sinai Sch Med, Div Liver Dis, New York, NY USA. Univ Calif San Francisco, San Francisco Gen Hosp, Dept Med, Div Pulm & Crit Care Med, San Francisco, CA USA. Univ Colorado, Dept Mol Cellular & Dev Biol, Boulder, CO 80309 USA. NCI, Ctr Canc Res, Cell & Canc Biol Branch, Bethesda, MD 20892 USA. FibroGen Inc, San Francisco, CA USA. Duke Univ, Dept Med, Durham, NC USA. Univ Florida, Dept Obstet & Gynecol, Gainesville, FL 32611 USA. Univ Minnesota, Dept Med Chem, Minneapolis, MN 55455 USA. NHLBI, Div Lung Dis, Bethesda, MD 20892 USA. RP Musson, RA (reprint author), Room 7126,MSC 7922,6701 Rockledge Dr, Bethesda, MD 20892 USA. OI LEINWAND, LESLIE/0000-0003-1470-4810 NR 84 TC 130 Z9 137 U1 1 U2 6 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1073-449X J9 AM J RESP CRIT CARE JI Am. J. Respir. Crit. Care Med. PD JUL 15 PY 2002 VL 166 IS 2 BP 236 EP 246 DI 10.1164/rccm.2201069 PG 11 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 571XX UT WOS:000176745400018 PM 12119236 ER PT J AU Evans, J Wang, TC Heyes, MP Markey, SP AF Evans, J Wang, TC Heyes, MP Markey, SP TI LC/MS analysis of NAD biosynthesis using stable isotope pyridine precursors SO ANALYTICAL BIOCHEMISTRY LA English DT Article ID PERFORMANCE LIQUID-CHROMATOGRAPHY; NICOTINIC-ACID; RAT-LIVER; REVERSED-PHASE; PURINE-BASES; NUCLEOTIDES; DINUCLEOTIDES; NUCLEOSIDES; TRYPTOPHAN; ADENINE AB A liquid chromatographic-electrospray ionization ion trap mass spectrometry (LUMS) method has been developed to measure the biosynthetic incorporation of specific precursors into NAD. The stable isotope-labeled precursors tryptophan, quinolinic acid, nicotinic acid, and nicotinamide were added to the media of human liver tumor cells (SK-HEP) grown in culture. The cells were harvested, the NAD was extracted, and the ratio of labeled to unlabeled NAD was measured using the newly developed LC/MS assay. The quantity of NAD formed from each precursor relative to an internal standard (fully labeled C-13, N-15-labeled NAD prepared from baker's yeast) was measured. The detection limit (signal-to-noise ratio 5:1) of the LC/MS method was 37 fmol (25 pg) of NAD and was linear from 20.0 ng to 25 pg. All reported NAD levels were normalized relative to cellular protein measurements. At 50 muM precursor concentrations, nicotinamide was the dominant precursor and NAD levels in the cell rose well above normal levels. Other precursors were minimally incorporated. The same methods were applied to NAD biosynthesized by macrophages derived from peripheral blood monocytes. However, the NAD concentration in macrophages was about 5% of that in SK-HEP cells and the incorporation of stable isotopelabeled substrates remained below measurable levels. C1 NIMH, Lab Neurotoxicol, Bethesda, MD 20892 USA. RP Evans, J (reprint author), NIMH, Lab Neurotoxicol, Bethesda, MD 20892 USA. NR 30 TC 11 Z9 12 U1 3 U2 8 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-2697 J9 ANAL BIOCHEM JI Anal. Biochem. PD JUL 15 PY 2002 VL 306 IS 2 BP 197 EP 203 DI 10.1006/abio.2002.5715 PG 7 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 574UL UT WOS:000176907900005 PM 12123656 ER PT J AU Chen, YR Mason, RP AF Chen, YR Mason, RP TI Mechanism in the reaction of cytochrome c oxidase with organic hydroperoxides: an ESR spin-trapping investigation SO BIOCHEMICAL JOURNAL LA English DT Article DE compound F; compound P; free radical; oxidative damage ID HYDROGEN-PEROXIDE; HEART-MITOCHONDRIA; SUPEROXIDE FORMATION; RADICAL FORMATION; 2.8 ANGSTROM; GENERATION; CYANYL; H2O2; METMYOGLOBIN; INVOLVEMENT AB Organic hydroperoxides are of great utility in probing the reaction mechanism and the toxicological consequences of lipid peroxidation. In the present study, ESR spin-trapping was employed to investigate the peroxidation of mitochondrial cytochrome c oxidase (CcO) with t-butyl hydroperoxide (t-BuOOH) and cumene hydroperoxide (CumOOH). The spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was used to detect the radical species formed from the reaction of CcO with t-BuOOH. The presence of t-BuOOH-derived alkoxyl radical (t-BuO.) as the primary radical indicates reductive scission of the O-O bond by CcO. The ESR signal of DMPO/(.)Ot-Bu can be partially abolished by cyanide, implying that the reductive cleavage involved the haem a(3)Cu(B) binuclear site of CcO. A nitroso spin trap, 2-methyl-2-nitrosopropane (MNP), was used to detect and identify radical species from the reaction of CcO with CumOOH. In addition to the t-BuOOH-derived methyl, hydroxylmethyl and tertiary carbon-centred radicals, a protein-derived radical was detected. The intensity of the ESR signal from the protein radical increased with the CumOOH concentration at low CumOOH/CcO ratios, with maximal intensity at a ratio of 100 mol of CumOOH/mol of CcO. The immobilized protein radical adduct of MNP was stable and persistent after dialysis; it was also resistant to proteolytic digestion, suggesting that it was formed in the transmembrane region, a region that is not accessible to proteases. Its signal was greatly enhanced when CeO cysteine residues were chemically modified by N-ethylmaleimide, when the tryptophan residues in CcO were oxidized by N-bromosuccimide, and when tyrosine residues on the surface of CcO were iodinated, showing that a radical equilibrium was established among the cysteine, tryptophan and tyrosine residues of the protein-centred radical. Pre-treatment of CcO with cyanide prevented detectable MNP adduct formation, confirming that the haem a(3)-Cu-B binuclear centre was the initial reaction site. When the CeO was pre-treated with 10 mM (100 equivalents) of CumOOH, the enzyme activity decreased by more than 20%. This inhibition was persistent after dialysis, suggesting that the detected protein-centred radical was, in part, involved in the irreversible inactivation by CumOOH. Visible spectroscopic analysis revealed that the haem a of CcO was not affected during the reaction. However, the addition of pyridine to the reaction mixture under alkaline conditions resulted in the destruction of the haem centre of CcO, suggesting that its protein matrix rather than its haem a is the target of oxidative damage by the organic hydroperoxide. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. RP Chen, YR (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 43 TC 19 Z9 19 U1 2 U2 7 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD JUL 15 PY 2002 VL 365 BP 461 EP 469 DI 10.1042/BJ20020170 PN 2 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577NC UT WOS:000177066400014 PM 11931642 ER PT J AU Fuda, H Shimizu, C Lee, YC Akita, H Strott, CA AF Fuda, H Shimizu, C Lee, YC Akita, H Strott, CA TI Characterization and expression of human bifunctional 3 '-phosphoadenosine 5 '-phosphosulphate synthase isoforms SO BIOCHEMICAL JOURNAL LA English DT Article DE cartilage; sulphate activation; sulphoconjugation; sulphonate donor; sulphonation ID ATP SULFURYLASE; SULFATE ACTIVATION; PAPS SYNTHETASE; KINASE; CLONING; MOUSE; SULFOTRANSFERASES; LOCALIZATION; PATHWAY; GENES AB Sulphonation, a fundamental process essential for normal growth and development, requires the sulphonate donor molecule 3'-phosphoadenosine 5'-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the bifunctional enzyme PAPS synthase. In humans, two genes encode isoenzymes that are 77% identical at the amino acid level, and alternative splicing creates two subtypes of PAPS synthase 2. The question as to whether distinctions in amino acid composition are reflected in differences in activity has been examined. The specific activity of the PAPS synthase 2 subtypes is 10- to 15-fold higher than that for PAPS synthase 1. The greater catalytic efficiency of the PAPS synthase 2 subtypes is demonstrated further by the 3- to 6-fold higher k(cat)/K-m ratios for ATP and inorganic sulphate as compared with the ratios for PAPS synthase 1. In humans, PAPS synthase I is expressed ubiquitously, and is the dominant isoform in most tissues, whereas expression of the PAPS synthase 2 subtypes is variable and tissue-specific. It is noteworthy that, similar to other human tissues, PAPS synthase 1 also appears to be the dominant isoform expressed in cartilage. The latter finding initially created a conundrum, since there is a specific human dwarfing disorder that is known to be caused by a mutation in the PAPS synthase 2 gene. This apparent enigma would seem to be resolved by examination of cartilage from guinea-pigs as an animal model. Similar to humans, cartilage from mature animals predominantly expresses PAPS synthase 1. In contrast, expression of PAPS synthase I is relatively low in the cartilage of immature guinea-pigs, including the growth plate of long bones, whereas PAPS synthase 2 is the highly expressed isoenzyme. C1 NICHHD, Sect Steroid Regulat, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Strott, CA (reprint author), NICHHD, Sect Steroid Regulat, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. NR 21 TC 33 Z9 34 U1 2 U2 4 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD JUL 15 PY 2002 VL 365 BP 497 EP 504 DI 10.1042/BJ20020044 PN 2 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 577NC UT WOS:000177066400018 PM 11931637 ER PT J AU Wang, CY Lei, HJ Huang, CYF Zhang, ZJ Mukherjee, AB Yuan, CJ AF Wang, CY Lei, HJ Huang, CYF Zhang, ZJ Mukherjee, AB Yuan, CJ TI Induction of cyclooxygenase-2 by staurosporine through the activation of nuclear factor for IL-6 (NF-IL6) and activator protein 2 (AP2) in an osteoblast-like cell line SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE staurosporine; NF-IL6; AP2; cyclooxygenase-2 ID PROSTAGLANDIN-ENDOPEROXIDE SYNTHASE-2; NECROSIS-FACTOR-ALPHA; FACTOR-KAPPA-B; KINASE-C; PHORBOL ESTER; TRANSCRIPTIONAL REGULATION; RETINOBLASTOMA PROTEIN; G(1) ARREST; C/EBP-BETA; GENE AB The induction of cyclooxygenase-2 (COX-2) plays a crucial role in many physiological and pathological processes. The expression of the COX-2 gene is regulated by many extracellular stimuli, including growth factors, cytokines, and tumor promoters. Staurosporine, a potential anti-tumor drug, was found recently to up-regulate the expression of the COX-2 gene in the mouse osteoblast-like cell line MC3T3-E1. The ability of staurosporine to induce the expression of the COX-2 gene was investigated using luciferase reporters controlled by various COX-2 core promoter regions. Two cis-acting sites for activator protein 2 (AP2) and nuclear factor for IL-6 (NF-IL6), respectively, were identified as responsible for the staurosporine-mediated COX-2 up-regulation. Mutational analysis further verified that both NF-IL6 and AP2 are involved in this process. Further studies showed the stimulatory effect of staurosporine on luciferase activity to be both time- and concentration-dependent. Luciferase activity could be induced at as low as 5 nM staurosporine and reached a maximum at around 20 nM. At 50 nM, the stimulatory effect of staurosporine on luciferase activity reached a maximum at about 8 hr and fell rapidly following 10 hr of incubation. Interestingly, a selective protein kinase C inhibitor, 2-[1-(3-dimethylaminopropyl)indol-3-yl]-3-(indol-3-yl) maleimide (GF109203X), failed to stimulate luciferase activity under the same conditions. This finding implies that staurosporine-mediated COX-2 gene expression is specific and independent of protein kinase C activity. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Natl Chiao Tung Univ, Dept Biol Sci & Technol, Hsinchu 30050, Taiwan. Natl Hlth Res Inst, Dept Mol Genet, Taipei, Taiwan. NICHD, NIH, Sect Dev Genet, Heritable Disorders Branch, Bethesda, MD 20892 USA. RP Yuan, CJ (reprint author), Natl Chiao Tung Univ, Dept Biol Sci & Technol, 75 Po Ai St, Hsinchu 30050, Taiwan. NR 43 TC 5 Z9 5 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD JUL 15 PY 2002 VL 64 IS 2 BP 177 EP 184 AR PII S0006-2952(02)01106-1 DI 10.1016/S0006-2952(02)01106-1 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 580KH UT WOS:000177233400003 PM 12123737 ER PT J AU Shaywitz, BA Shaywitz, SE Pugh, KR Mencl, WE Fulbright, RK Skudlarski, P Constable, RT Marchione, KE Fletcher, JM Lyon, GR Gore, JC AF Shaywitz, BA Shaywitz, SE Pugh, KR Mencl, WE Fulbright, RK Skudlarski, P Constable, RT Marchione, KE Fletcher, JM Lyon, GR Gore, JC TI Disruption of posterior brain systems for reading in children with developmental dyslexia SO BIOLOGICAL PSYCHIATRY LA English DT Article DE dyslexia; reading; fMRI; children; brain ID LOW ACHIEVEMENT DEFINITIONS; FUNCTIONAL CONNECTIVITY; CEREBRAL MECHANISMS; PHONOLOGICAL-CORE; ANGULAR GYRUS; WORD; DISABILITY; DISCREPANCY; PERFORMANCE; PROFILES AB Background: Converging evidence indicates a functional disruption in the neural systems,for reading in adults with dyslexia. We examined brain activation patterns in dyslexic and nonimpaired children during pseudoword and real-word reading tasks that required phonologic analysis (i.e., tapped the problems experienced by dyslexic children in sounding out words). Methods: We used functional magnetic resonance imaging (fMRI) to study 144 right-handed children, 70 dyslexic readers, and 74 nonimpaired readers as they read pseudowords and real words. Results: Children with dyslexia demonstrated a disruption in neural systems for reading involving posterior brain regions, including parietotemporal sites and sites in the occipitotemporal area. Reading skill was positively correlated with the magnitude of activation in the left occipitotemporal region. Activation in the left and right inferior frontal gyri was greater in older compared with younger dyslexic children. Conclusions: These findings provide neurobiological evidence of an underlying disruption in the neural systems for reading in children with dyslexia and indicate that it is evident at a young age. The locus of the disruption places childhood dyslexia within the same neurobiological framework as dyslexia, and acquired alexia, occurring in adults. C1 Yale Univ, Sch Med, Dept Pediat, New Haven, CT 06510 USA. Yale Univ, Sch Med, Dept Diagnost Radiol, New Haven, CT 06510 USA. Yale Univ, Sch Med, Dept Appl Phys, New Haven, CT 06510 USA. Haskins Labs Inc, New Haven, CT 06511 USA. Univ Texas, Sch Med, Dept Pediat, Houston, TX USA. NICHD, Child Dev & Behav Branch, NIH, Washington, DC USA. RP Shaywitz, BA (reprint author), Yale Univ, Sch Med, Dept Pediat, POB 3333, New Haven, CT 06510 USA. FU NICHD NIH HHS [P01 HD 21888, P50 HD25802] NR 53 TC 497 Z9 505 U1 6 U2 46 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD JUL 15 PY 2002 VL 52 IS 2 BP 101 EP 110 AR PII S0006-3223(02)01365-3 DI 10.1016/S0006-3223(02)01365-3 PG 10 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 572LY UT WOS:000176777200004 PM 12114001 ER PT J AU Lee, SJ Klein, JP Barrett, AJ Ringden, O Antin, JH Cahn, JY Carabasi, MH Gale, RP Giralt, S Hale, GA Ilhan, O McCarthy, PL Socie, G Verdonck, LF Weisdorf, DJ Horowitz, MM AF Lee, SJ Klein, JP Barrett, AJ Ringden, O Antin, JH Cahn, JY Carabasi, MH Gale, RP Giralt, S Hale, GA Ilhan, O McCarthy, PL Socie, G Verdonck, LF Weisdorf, DJ Horowitz, MM TI Severity of chronic graft-versus-host disease: association with treatment-related mortality and relapse SO BLOOD LA English DT Article ID BONE-MARROW-TRANSPLANTATION; CHRONIC MYELOGENOUS LEUKEMIA; BLOOD PROGENITOR-CELL; CHRONIC PHASE; HIGH-RISK; SURVIVAL; CYCLOSPORINE; PREDNISONE; PREDICTORS; DONOR AB Chronic graft-versus-host disease (cGVHD) is the leading cause of late treatment-related deaths among recipients of allogeneic bone marrow and blood transplants. However, cGVHD is also associated with fewer relapses. We sought to determine whether severity of cGVHD predicts the magnitude of these effects. One impediment to such an analysis is the current limited/extensive grading system for cGVHD because this classification was designed to identify patients likely to benefit from systemic immune suppression and does not capture the severity of multiorgan involvement. We, therefore, first developed a grading system predictive for survival by using data from 1827 HLA-matched sibling allotransplant recipients reported to the International Bone Marrow Transplant Registry (IBMTR). We found Karnofsky performance score, diarrhea, weight loss, and cutaneous and oral involvement to be independent prognostic variables, from which we generated a grading scheme. We tested this scheme, the limited/extensive classification system, and a classification based on clinical impression of overall cGVHD severity (mild/moderate/severe) in parallel analyses of 1092 HLA-matched sibling transplant recipients from the IBMTR and 553 recipients of unrelated donor marrow from the National Marrow Donor Program. Presence of cGVHD was associated with fewer relapses (relative risk [RR], 0.5-0.6) but more treatment-related mortality (RR, 1.8-2.8) in the 3 analyses. No grading scheme correlated cGVHD severity with relapse rates, but all schemes predicted treatment-related mortality. Survival and disease-free survival of the most favorable cGVHD group in each scheme were similar, or better, than those of patients without cGVHD; these patients may not need aggressive or extended immune suppression. (Blood. 2002; 100:406-414). (C) 2002 by The American Society of Hematology. C1 Med Coll Wisconsin, Graft Vs Host Dis Working Comm Int Bone Marrow Tr, Hlth Policy Inst, Milwaukee, WI 53226 USA. Natl Marrow Donor Program, Coordinating Ctr, Minneapolis, MN USA. Dana Farber Canc Ctr, Boston, MA USA. NHLBI, Bethesda, MD 20892 USA. Huddinge Hosp, S-14186 Huddinge, Sweden. Hop Jean Minjoz, F-25030 Besancon, France. Univ So Calif, Norris Canc Ctr, Los Angeles, CA USA. Ctr Adv Studies Leukemia, Los Angeles, CA USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. St Jude Childrens Res Hosp, Memphis, TN 38105 USA. Ibni Sina Hosp, Ankara, Turkey. Roswell Pk Canc Inst, Buffalo, NY 14263 USA. Hop St Louis, Paris, France. Univ Utrecht Hosp, Utrecht, Netherlands. Fairview Univ, Med Ctr, Minneapolis, MN USA. RP Lee, SJ (reprint author), Dana Farber Canc Inst, 44 Binney St, Boston, MA 02115 USA. EM stephanie_lee@dfci.harvard.edu FU NCI NIH HHS [CA75267-03, P01-CA-40053]; PHS HHS [U24-76518] NR 33 TC 271 Z9 284 U1 2 U2 6 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD JUL 15 PY 2002 VL 100 IS 2 BP 406 EP 414 DI 10.1182/blood.V100.2.406 PG 9 WC Hematology SC Hematology GA 571WB UT WOS:000176741200005 PM 12091329 ER PT J AU Yavuz, AS Lipsky, PE Yavuz, S Metcalfe, DD Akin, C AF Yavuz, AS Lipsky, PE Yavuz, S Metcalfe, DD Akin, C TI Evidence for the involvement of a hematopoietic progenitor cell in systemic mastocytosis from single-cell analysis of mutations in the c-kit gene SO BLOOD LA English DT Article ID B-CELLS; HEMATOLOGIC DISORDER; CATALYTIC DOMAIN; MYELOID CELLS; IDENTIFICATION; CHROMOSOME; LEUKEMIA; BURDEN; BLOOD; LINE AB Mast cells are derived from multipotential hernatopoietic progenitors and are clonally increased in systemic mastocytosis, a disease associated with point mutations of codon 816 (most commonly Asp816Val) of c-kit. To study the lineage relationship and the extent of expansion of cells derived from the mutated clone, we examined the occurrence of the Asp816Val c-kit mutation in genomic DNA of individual sorted peripheral blood T cells, B cells, and monocytes in patients with indolent systemic mastocytosis. The mutation was detected in varying frequencies in the genomic DNA of individual B cells and monocytes and bone marrow mast cells in patients with extensive disease. In B cells, the immunoglobulin repertoire was polyclonal, indicating that the mutation occurred before V-H/(D)/J(H) recombination. These results show that mastocytosis is a disorder of a pluripotential hematopoietic progenitor cell that gives rise to B cells and monocytes in addition to mast cells and that the affected clone shows variable expansion in these lineages in the peripheral blood of patients with systemic mastocytosis. (C) 2002 by The American Society of Hematology. C1 NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. NIAMS, Autoimmun Branch, Bethesda, MD USA. RP Akin, C (reprint author), NIAID, Lab Allerg Dis, NIH, 10 Ctr Dr,Rm 11C205, Bethesda, MD 20892 USA. OI Akin, Cem/0000-0001-6301-4520 NR 17 TC 115 Z9 121 U1 0 U2 2 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD JUL 15 PY 2002 VL 100 IS 2 BP 661 EP 665 DI 10.1182/blood-2002-01-0203 PG 5 WC Hematology SC Hematology GA 571WB UT WOS:000176741200038 PM 12091362 ER PT J AU Micheli, A Yancik, R Krogh, V Verdecchia, A Sant, M Capocaccia, R Berrino, F Ries, L AF Micheli, A Yancik, R Krogh, V Verdecchia, A Sant, M Capocaccia, R Berrino, F Ries, L TI Contrasts in cancer prevalence in Connecticut, Iowa, and Utah SO CANCER LA English DT Article DE cancer prevalence; stomach carcinoma; colorectal carcinoma; pancreatic carcinoma; breast carcinoma; carcinoma of the corpus uteri; ovarian carcinoma; non-Hodgkin lymphoma ID CHRONIC DISEASE MORBIDITY; TUMOR-REGISTRY; BREAST-CANCER; COMORBIDITY; MORTALITY; PROGRAM AB BACKGROUND. Cancer prevalence-the proportion of a population with cancer, including those recently diagnosed, those in treatment, and survivors-is an important indicator of future health care requirements. Only limited information on cancer prevalence is available for the United States. In particular, comparative interstate studies are not available. In this study, we estimate and analyze the prevalence of seven major cancers in Connecticut, Iowa, and Utah using the tried and tested PREVAL method applied to National Cancer Institute registry data. METHODS. We analyzed data on 242,851 carcinomas of the stomach, colorectum, pancreas, breast, uterus (corpus), ovary, and non-Hodgkin lymphoma (NHL), diagnosed in white Americans from 1973 through 1992. Observed prevalence was estimated by applying the PREVAL method to incidence and life status data from the cancer registries. Complete prevalence was estimated by applying correction factors obtained by modeling incidence and survival rates. RESULTS. The ratio of the highest to the lowest prevalence (as proportions) ranged from 1.69 for uterine carcinoma to 2.73 for stomach carcinoma, showing that marked differences in cancer prevalence exist within the United States. Utah had the lowest prevalence for each carcinoma. Connecticut and Iowa had similar prevalence levels for carcinomas of the colorectum, pancreas, and ovary and for NHL. Breast carcinoma was the most prevalent, with 826 cases per 100,000 of population in Utah, 1518 per 100,000 in Iowa, and 1619 per 100,000 in Connecticut. Cancer survival did not differ greatly among the three registry populations. The major determinants of prevalence differences were incidence and the population age distribution. CONCLUSIONS. PREVAL provides reliable estimates of the numbers of living people in a population who have had a cancer diagnosis. Prevalence depends on incidence and survival and on the age structure of population. All these factors have changed markedly in recent years and will continue to do so in the future. Cancer prevalence should be monitored over time to evaluate changes by area, sex, age, and cancer site. The prevalence figures presented are directly comparable with those from European cancer registries. (C) 2002 American Cancer Society. C1 Ist Nazl Studio & Cura Tumori, Unita Epidemiol, I-20133 Milan, Italy. NIA, Bethesda, MD 20892 USA. Ist Super Sanita, Epidemiol & Biostat Lab, I-00161 Rome, Italy. NCI, Canc Stat Branch, SEER Program, Bethesda, MD 20892 USA. RP Micheli, A (reprint author), Ist Nazl Studio & Cura Tumori, Unita Epidemiol, Via Venezian 1, I-20133 Milan, Italy. RI Krogh, Vittorio/K-2628-2016; Sant, Milena/D-2362-2017; OI Krogh, Vittorio/0000-0003-0122-8624; Sant, Milena/0000-0002-4148-8597; Micheli, Andrea/0000-0002-4558-4754 NR 21 TC 6 Z9 6 U1 1 U2 1 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0008-543X J9 CANCER JI Cancer PD JUL 15 PY 2002 VL 95 IS 2 BP 430 EP 439 DI 10.1002/cncr.10675 PG 10 WC Oncology SC Oncology GA 575CH UT WOS:000176928100029 PM 12124844 ER PT J AU Calvo, A Feldman, AL Libutti, SK Green, JE AF Calvo, A Feldman, AL Libutti, SK Green, JE TI Adenovirus-mediated endostatin delivery results in inhibition of mammary gland tumor growth in C3(1)/SV40 T-antigen transgenic mice SO CANCER RESEARCH LA English DT Article ID ANTIANGIOGENIC GENE-THERAPY; ANTITUMOR-ACTIVITY; MOUSE MODEL; ANGIOGENESIS; CANCER; METASTASES AB We demonstrate the efficacy of systemic administration of a replication-defective adenovirus expressing endostatin (Ad-mEndo) administered during the preinvasive stage of mammary tumor development in C3(1)/T antigen transgenic mice. Mean serum levels of endostatin increased about 8-fold above that of controls and resulted in a significant decrease in tumor growth and an increase in survival. The inhibitory effect of endostatin occurred during or after the progression to invasive carcinoma. Reduced levels of vascular endothelial growth factor mRNA were found in association with high levels of endostatin. Our results demonstrate that the adenoviral induction of high levels of circulating endostatin significantly inhibits mammary tumor growth during the period when the "angiogenic switch" occurs. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Green, JE (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bldg 41,Room C629,41 Lib Dr, Bethesda, MD 20892 USA. RI Feldman, Andrew/D-5028-2012 NR 20 TC 29 Z9 30 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUL 15 PY 2002 VL 62 IS 14 BP 3934 EP 3938 PG 5 WC Oncology SC Oncology GA 574CU UT WOS:000176871500008 PM 12124322 ER PT J AU Fears, TR Bird, CC Guerry, D Sagebiel, RW Gail, MH Elder, DE Halpern, A Holly, EA Hartge, P Tucker, MA AF Fears, TR Bird, CC Guerry, D Sagebiel, RW Gail, MH Elder, DE Halpern, A Holly, EA Hartge, P Tucker, MA TI Average midrange ultraviolet radiation flux and time outdoors predict melanoma risk SO CANCER RESEARCH LA English DT Article ID CUTANEOUS MALIGNANT-MELANOMA; SUN EXPOSURE; DYSPLASTIC NEVI; MORTALITY-RATES; UNITED-STATES AB Sunlight is the major environmental risk factor for melanoma. Descriptive studies have shown latitudinal variation in population incidence and mortality rates [D. C. Whiteman and A. C. Green, Int. J. Dermatol., 38: 481-489, 1999, and B. K. Armstrong, Australian J. Dermatol., 38 (Suppl. 1): 51-56, 1997]. In analytic studies, individual exposure has been particularly difficult to quantify. Lifetime residential history was coupled with levels of midrange UV radiation (UVB flux) to provide a measure of individual exposure to sunlight thought to be less subject to misclassification and recall bias. Data were analyzed from 718 non-Hispanic white patients with invasive cutaneous melanoma from melanoma clinics in Philadelphia and San Francisco. Matched controls were 945 patients from outpatient clinics with similar catchment areas. The association of melanoma risk and history of UVB flux along with the usual outdoor exposure risk factors were studied. A 10% increase in the average annual UVB flux was associated with a 19% [95% confidence interval (CI), 5-35%] increase in individual odds for melanoma for men and 16% (95% CI, 2-32%) for women. In men, a 10% increase in hours outdoors was associated with a 2.8% (95% CI, 1.2-4.5%) increase in odds. Even in women who could develop a deep tan, a 10% increase in hours outdoors was associated with a 5.8% increase in odds (95% CI, 1.4-10.4%). The association between melanoma risk and average annual UVB flux was strong and consistent for men and for women. The association with total adult hours outdoors was notable for men of all skin types and women who develop a suntan. C1 NCI, Div Canc Epidemiol & Genet, NIH, Bethesda, MD 20892 USA. Univ Penn, Sch Med, Pigmented Les Study Grp, Philadelphia, PA 19104 USA. Univ Calif San Francisco, Melanoma Clin, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Epidemiol & Biostat, San Francisco, CA 94143 USA. RP Fears, TR (reprint author), NCI, Div Canc Epidemiol & Genet, NIH, Execut Plaza S,Room 8040, Bethesda, MD 20892 USA. NR 19 TC 84 Z9 87 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUL 15 PY 2002 VL 62 IS 14 BP 3992 EP 3996 PG 5 WC Oncology SC Oncology GA 574CU UT WOS:000176871500018 PM 12124332 ER PT J AU Ojo-Amaize, EA Nchekwube, EJ Cottam, HB Bai, RL Verdier-Pinard, P Kakkanaiah, VN Varner, JA Leoni, L Okogun, JI Adesomoju, AA Oyemade, OA Hamel, E AF Ojo-Amaize, EA Nchekwube, EJ Cottam, HB Bai, RL Verdier-Pinard, P Kakkanaiah, VN Varner, JA Leoni, L Okogun, JI Adesomoju, AA Oyemade, OA Hamel, E TI Hypoestoxide, a natural nonmutagenic diterpenoid with antiangiogenic and antitumor activity: Possible mechanisms of action SO CANCER RESEARCH LA English DT Article ID ENDOTHELIAL GROWTH-FACTOR; HUMAN OVARIAN; CANCER CELLS; IN-VIVO; ANGIOGENESIS; METASTASIS; POLYMERIZATION; PROLIFERATION; INTERLEUKIN-8; CYTOKINESIS AB We have shown previously that hypoestoxide (HE), a natural diterpenoid [a bicyclo (9, 3, 1) pentadecane], is a potent nonsteroidal anti-inflammatory drug. In this report, we demonstrate that HE also inhibits the growth of a variety of human and murine tumor cell lines in vitro at concentrations ranging from 0.3 to 10 muM and was inactive as a mutagen in the Ames test. HE exhibited highly potent (0.3-10 mg/kg dose ranges) activities against B16 melanoma growth in C57BL/6 mice and P388D1 leukemia in C57BL/6 x DBA/2 F-I mice, respectively. At a low maximal effective dose of 5 mg/kg, HE induced significant in vivo antitumor activities that were better than or comparable with most of the standard chemotherapeutic antiangiogenic agents tested: cortisone acetate, vincris-tine, bleomycin, Adriamycin, 5-fluorouracil, cyclophosphamide, and etoposide. All of the agents, except vincristine, had much higher maximal effective doses than HE. HE arrested the growth of human Burkitt lymphoma CA46 cells and HeLa (cervical epitheloid carcinoma) cells in the G(2)-M phase of the cell cycle, which was caused by interference, either direct or indirect, with actin assembly. Thus, the cell cycle arrest occurred at cytokinesis, as demonstrated by an increase in the number of binucleate cells. Moreover, HE inhibited vascular endothelial growth factor-induced cell proliferation in vitro, with an IC50 of 28.6 muM, and it significantly inhibited basic fibroblast growth factor-induced angiogenesis on the chick chorioallantoic membrane, with an IC50 of 10 muM. Furthermore, HE inhibited endothelial cell migration on vitronectin, collagen, and fibronectin. Besides its activity as a nonsteroidal anti-inflammatory drug, HE also has promise for the chemotherapy of cancer. C1 Immune Modulat Inc, Bloomington, CA 92316 USA. Paraquest Inc, Bloomington, CA 92316 USA. NCI, Screening Technol Branch, DTP, DCTD,NIH, Frederick, MD 21702 USA. Univ Calif San Diego, Ctr Canc, La Jolla, CA 92093 USA. Univ Calif San Diego, Dept Med, La Jolla, CA 92093 USA. Univ Ibadan, Dept Chem, Ibadan, Nigeria. RP Ojo-Amaize, EA (reprint author), Immune Modulat Inc, POB 998, Bloomington, CA 92316 USA. OI Varner, Judith/0000-0002-9251-0600 NR 40 TC 15 Z9 16 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUL 15 PY 2002 VL 62 IS 14 BP 4007 EP 4014 PG 8 WC Oncology SC Oncology GA 574CU UT WOS:000176871500020 PM 12124334 ER PT J AU Turner, BC Ottey, M Zimonjic, DB Potoczek, M Hauck, WW Pequignot, E Keck-Waggoner, CL Sevignani, C Aldaz, CM McCue, PA Palazzo, J Huebner, K Popescu, NC AF Turner, BC Ottey, M Zimonjic, DB Potoczek, M Hauck, WW Pequignot, E Keck-Waggoner, CL Sevignani, C Aldaz, CM McCue, PA Palazzo, J Huebner, K Popescu, NC TI The fragile histidine triad/common chromosome fragile site 3B locus and repair-deficient cancers SO CANCER RESEARCH LA English DT Article ID BREAST-CANCER; FHIT EXPRESSION; GENE-MUTATIONS; DNA-REPAIR; CARCINOMAS; BRCA1; SUPPRESSOR; WWOX; APHIDICOLIN; PHENOTYPE AB In various studies of sporadic breast cancers, 40-70% were strongly positive for fragile histidine triad (Fhit) protein expression, whereas only 18% of BRCA2 mutant breast cancers demonstrated strong Fhit expression, suggesting that the BRCA2 repair function may be necessary to retain intact fragile common chromosome fragile site 3B (FRA3B)/FHIT loci. In the current study, 22 breast tumors with deleterious BRCA1 mutations were analyzed for Fhit expression by immunohistochemistry in a case-control matched pair analysis. Loss of Fhit expression was significantly more frequent in the BRCA1 cancers compared with sporadic breast tumors (9% Fhit positive versus 68% Fhit positive), suggesting that the BRCA1 pathway is also important in protecting the FRA3B/FHIT locus from damage. To investigate the relationship between repair gene deficiencies and induction of chromosome fragile sites in vitro, we have analyzed the frequency of aphidicolin induction of chromosome gaps and breaks in PMS2-, BRCA1-, MSH2-, MLH1-, FHIT-, and TP53-deficient cell lines. Each of the repair-deficient cell lines showed elevated expression of chromosome gaps and breaks, consistent with the proposal that proteins involved in mismatch and double-strand break repair are important in maintaining the integrity of common fragile regions. Correspondingly, genes at common fragile sites may sustain elevated levels of DNA damage in cells with deficient DNA repair proteins such as those mutated in several familial cancer syndromes. C1 Thomas Jefferson Univ, Kimmel Canc Ctr, BLSB, Dept Radiat Oncol, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Kimmel Canc Ctr, Dept Microbiol & Immunol, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Kimmel Canc Ctr, Dept Med, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Kimmel Canc Ctr, Dept Pathol, Philadelphia, PA 19107 USA. NCI, Expt Carcinogenesis Lab, Bethesda, MD 20892 USA. Univ Texas, MD Anderson Canc Ctr, Dept Carcinogenesis, Div Sci Pk Res, Smithville, TX 77030 USA. RP Huebner, K (reprint author), Thomas Jefferson Univ, Kimmel Canc Ctr, BLSB, Dept Radiat Oncol, Room 1008,233 S 10th St, Philadelphia, PA 19107 USA. NR 39 TC 48 Z9 49 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUL 15 PY 2002 VL 62 IS 14 BP 4054 EP 4060 PG 7 WC Oncology SC Oncology GA 574CU UT WOS:000176871500027 PM 12124341 ER PT J AU Moss, WJ Monze, M Ryon, JJ Quinn, TC Griffin, DE Cutts, F AF Moss, WJ Monze, M Ryon, JJ Quinn, TC Griffin, DE Cutts, F TI Prospective study of measles in hospitalized, human immunodeficiency virus (HIV)-infected and HIV-uninfected children in Zambia SO CLINICAL INFECTIOUS DISEASES LA English DT Article ID INFECTED CHILDREN; IMMUNITY; VACCINATION AB Measles in persons coinfected with human immunodeficiency virus (HIV) has been reported to be unusual in its presentation and frequently fatal. To determine the effect of HIV coinfection on the clinical features and outcome of measles, a prospective study of hospitalized children with measles was conducted between January 1998 and October 2000 in Lusaka, Zambia. One-sixth (17%) of 546 children hospitalized with laboratory-confirmed measles were coinfected with HIV. One-third of the HIV-infected children hospitalized with confirmed measles were <9 months old, compared with 23% of HIV-uninfected children (P=.03). Few differences in clinical manifestations, complications, or mortality were found between HIV-infected and HIV-uninfected children with measles. HIV-infected children constitute a significant proportion of children hospitalized with measles in countries with high HIV prevalence and are more likely to be younger than the age for routine measles immunization. C1 Johns Hopkins Univ, Bloomberg Sch Publ Hlth, W Harry Feinstone Dept Mol Microbiol & Immunol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Int Hlth, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Div Infect Dis, Baltimore, MD 21205 USA. NIAID, NIH, Bethesda, MD 20892 USA. Univ Teaching Hosp, Virol Lab, Lusaka, Zambia. London Sch Hyg & Trop Med, Dept Infect & Trop Dis, London, England. RP Moss, WJ (reprint author), Johns Hopkins Univ, Bloomberg Sch Publ Hlth, W Harry Feinstone Dept Mol Microbiol & Immunol, 515 N Wolfe St, Baltimore, MD 21205 USA. FU NIAID NIH HHS [AI-23047] NR 24 TC 46 Z9 48 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD JUL 15 PY 2002 VL 35 IS 2 BP 189 EP 196 DI 10.1086/341248 PG 8 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 567FQ UT WOS:000176475100011 PM 12087526 ER PT J AU Dass, KN Smith, MA Gill, VJ Goldstein, SA Lucey, DR AF Dass, KN Smith, MA Gill, VJ Goldstein, SA Lucey, DR TI Brevibacterium endocarditis: A first report SO CLINICAL INFECTIOUS DISEASES LA English DT Article ID IMMUNOCOMPROMISED PATIENT; PERITONITIS; BACTEREMIA; OTITIDIS; CASEI AB There are few case reports of infections caused by Brevibacterium species, and there have been no previously reported cases of endocarditis caused by any of the 6 known species of Brevibacterium. We report the first case of Brevibacterium endocarditis (caused by Brevibacterium otitidis) in a patient with prosthetic heart valves. The patient responded to 6 weeks of treatment with vancomycin and 2 weeks with gentamicin, and she has been receiving long-term maintenance therapy with oral azithromycin. C1 Washington Hosp Ctr, Infect Dis Serv, Washington, DC 20010 USA. Washington Hosp Ctr, Serv Cardiol, Washington, DC 20010 USA. NIH, Microbiol Serv, Dept Clin Pathol, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Dass, KN (reprint author), Washington Hosp Ctr, Infect Dis Serv, Rm 2A-56, Washington, DC 20010 USA. NR 10 TC 10 Z9 10 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD JUL 15 PY 2002 VL 35 IS 2 BP E20 EP E21 DI 10.1086/340984 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 567FQ UT WOS:000176475100024 PM 12087539 ER PT J AU Brody, T Stivers, C Odenwald, WF AF Brody, T Stivers, C Odenwald, WF TI A search for targets of the drosophila neuroblast temporal network. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract ID CNS C1 NINDS, Neurogenet Unit, Neurochem Lab, NIH, Bethesda, MD USA. NR 3 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 115 BP 464 EP 464 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700117 ER PT J AU Kishigami, S Trisha, C Mishina, Y AF Kishigami, S Trisha, C Mishina, Y TI BMP signaling through ALK2 plays a role as tight determinant in the establishment of left-right asymmetry in mouse embryos. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 NIEHS, Reproduct & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 125 BP 466 EP 466 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700126 ER PT J AU Yamaguchi, TP Biris, K Greear, J AF Yamaguchi, TP Biris, K Greear, J TI Wnt3a is required for establishment of L-R asymmetry. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 NCI, NIH, Frederick, MD 21701 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 124 BP 466 EP 466 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700125 ER PT J AU Itoh, M Dorsky, RI Moon, RT Chitnis, A AF Itoh, M Dorsky, RI Moon, RT Chitnis, A TI Basal repression of writ target genes by hdl and tcf3b helps define the low end of a writ activity gradient in the neurectoderm. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 NICHD, LMG, NIH, Bethesda, MD 20892 USA. Univ Utah, Dept Neurobiol & Anat, Salt Lake City, UT 84132 USA. Univ Washington, Howard Hughes Med Inst, Dept Pharmacol, Seattle, WA 98195 USA. Univ Washington, Ctr Dev Biol, Seattle, WA 98195 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 140 BP 468 EP 469 PG 2 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700136 ER PT J AU Habas, R Kato, Y Dawid, I He, X AF Habas, R Kato, Y Dawid, I He, X TI The role of the Rho family of GTPases in gastrulation cell movements. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 Harvard Univ, Childrens Hosp, Sch Med, Dept Neurol,Div Neurosci, Boston, MA 02115 USA. NICHD, Mol Genet Lab, NIH, Bethesda, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 160 BP 473 EP 473 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700155 ER PT J AU Voronina, VA Kozlov, SV Mathers, PH Lewandoski, M AF Voronina, VA Kozlov, SV Mathers, PH Lewandoski, M TI Conditional inactivation of the Rx homeobox gene results in viable anophthalmic animals. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 W Virginia Univ, Sensory Neurosci Res Ctr, Morgantown, WV 26506 USA. NCI, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 241 BP 490 EP 491 PG 2 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700226 ER PT J AU Waters, ST Wilson, C Anderson, R Lewandoski, M AF Waters, ST Wilson, C Anderson, R Lewandoski, M TI Analysis of GBX genes during neurogenesis. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 NCI, Frederick Canc Res & Dev Ctr, CCR, CDBL,NIH, Frederick, MD 21702 USA. SAIC Frederick, Frederick, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 242 BP 491 EP 491 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700227 ER PT J AU Kuzin, A Stivers, C Brody, T Odenwald, WF AF Kuzin, A Stivers, C Brody, T Odenwald, WF TI nerfin-1, a member of a conserved Zn-finger gene subfamily, is required for proper neuronal cell fate specification. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 Neurogenet Unit, LNC, NINDS, NIH, Bethesda, MD USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 285 BP 501 EP 501 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700267 ER PT J AU Miura, S Tallquist, MD Soriano, P Mishina, Y AF Miura, S Tallquist, MD Soriano, P Mishina, Y TI BMP signaling is important for mesoderm induction and germ layer development in mouse embryogenesis. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 NIEHS, LRDT, Mol Dev Biol Grp, NIH, Res Triangle Pk, NC 27709 USA. UT SW Med Ctr, Dept Mol Biol, Dallas, TX 75390 USA. Fred Hutchinson Canc Res Ctr, Program Dev Biol, Seattle, WA 98109 USA. Fred Hutchinson Canc Res Ctr, Div Basic Sci, Seattle, WA 98109 USA. RI Soriano, Philippe/E-5797-2015 OI Soriano, Philippe/0000-0002-0427-926X NR 0 TC 3 Z9 3 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 309 BP 505 EP 505 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700285 ER PT J AU Hoffman, MP Kidder, BL Steinberg, Z Larsen, M Kleinman, HK AF Hoffman, MP Kidder, BL Steinberg, Z Larsen, M Kleinman, HK TI Fibroblast growth factor receptors-1 and-2(IIIb), FGF7, and FGF10 regulate branching morphogenesis of developing mouse submandibular glands in organ culture. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 CDBRB, NIDCR, NIH, Bethesda, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 315 BP 507 EP 507 PG 1 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700291 ER PT J AU Stottmann, RW Mishina, Y Klingensmith, JA AF Stottmann, RW Mishina, Y Klingensmith, JA TI BMP signaling in mammalian neural tube development. SO DEVELOPMENTAL BIOLOGY LA English DT Meeting Abstract C1 Duke Univ, Ctr Med, Dept Cell Biol, Durham, NC USA. NIEHS, Res Triangle Pk, NC USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2002 VL 247 IS 2 MA 345 BP 513 EP 514 PG 2 WC Developmental Biology SC Developmental Biology GA 573LG UT WOS:000176830700318 ER PT J AU Kim, Y Yun, CW Philpott, CC AF Kim, Y Yun, CW Philpott, CC TI Ferrichrome induces endosome to plasma membrane cycling of the ferrichrome transporter, Arn1p, in Saccharomyces cerevisiae SO EMBO JOURNAL LA English DT Article DE endocytosis; exocytosis; iron; siderophore; yeast ID MAJOR FACILITATOR SUPERFAMILY; MICROBIAL IRON TRANSPORT; GPI-ANCHORED PROTEINS; VACUOLAR HYDROLASES; SIDEROPHORE-IRON; YEAST; ENDOCYTOSIS; GOLGI; IDENTIFICATION; DELIVERY AB Siderophores are small iron-binding molecules that are synthesized and secreted in the iron-free form by microorganisms. Saccharomyces cerevisiae takes up iron bound to siderophores by two separate systems, one of which requires the ARN family of sidero phore-iron transporters. Arn1p and Arn3p are expressed in endosome-like intracellular vesicles. Here we present evidence that, in the absence of its specific substrate, ferrichrome, Arn1p is sorted directly from the Golgi to the endosomal compartment and does not cycle to the plasma membrane. When cells are exposed to ferrichrome at low concentrations, Arn1p stably relocalizes to the plasma membrane. At higher concentrations of ferrichrome, Arn1p relocalizes to the plasma membrane and rapidly undergoes endocytosis. Plasma membrane localization of Arn1p occurs only in the presence of its specific substrate, and not in the presence of other siderophores. Despite expression of Arn1p on the plasma membrane, mutant strains with defects in endocytosis exhibit reduced uptake of ferrichrome-iron. Thus, siderophores influence the trafficking of the Arn transporters within the cell and this trafficking is important for transporter function. C1 NIDDK, Liver Dis Sect, NIH, Bethesda, MD 20892 USA. RP NIDDK, Liver Dis Sect, NIH, Bldg 10,Room 9B16,10 Ctr Dr, Bethesda, MD 20892 USA. EM carolinep@intra.niddk.nih.gov NR 48 TC 47 Z9 47 U1 0 U2 1 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0261-4189 EI 1460-2075 J9 EMBO J JI Embo J. PD JUL 15 PY 2002 VL 21 IS 14 BP 3632 EP 3642 DI 10.1093/emboj/cdf382 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 572PT UT WOS:000176784700008 PM 12110576 ER PT J AU Novoselov, SV Rao, M Onoshko, NV Zhi, HJ Kryukov, GV Xiang, YB Weeks, DP Hatfield, DL Gladyshev, VN AF Novoselov, SV Rao, M Onoshko, NV Zhi, HJ Kryukov, GV Xiang, YB Weeks, DP Hatfield, DL Gladyshev, VN TI Selenoproteins and selenocysteine insertion system in the model plant cell system, Chlamydomonas reinhardtii SO EMBO JOURNAL LA English DT Article DE Chlamydomonas reinhardtii; evolutionary analysis; SECIS elements; selenoproteins ID METHIONINE SULFOXIDE REDUCTASE; THIOREDOXIN REDUCTASE; CONTAINING PROTEINS; ESCHERICHIA-COLI; SECIS ELEMENTS; GENE; IDENTIFICATION; EXPRESSION; DROSOPHILA; SEQUENCE AB Known eukaryotic selenocysteine (Sec)-containing proteins are animal proteins, whereas selenoproteins have not been found in yeast and plants. Surprisingly, we detected selenoproteins in a member of the plant kingdom, Chlamydomonas reinhardtii, and directly identified two of them as phospholipid hydroperoxide glutathione peroxidase and selenoprotein W homologs. Moreover, a selenocysteyl-tRNA was isolated that recognized specifically the Sec codon UGA. Subsequent gene cloning and bioinformatics analyses identified eight additional selenoproteins, including methionine-S-sulfoxide reductase, a selenoprotein specific to Chlamydomonas. Chlamydomonas selenoprotein genes contained selenocysteine insertion sequence (SECIS) elements that were similar, but not identical, to those of animals. These SECIS elements could direct selenoprotein synthesis in mammalian cells, indicating a common origin of plant and animal Sec insertion systems. We found that selenium is required for optimal growth of Chlamydomonas. Finally, evolutionary analyses suggested that selenoproteins present in Chlamydomonas and animals evolved early, and were independently lost in land plants, yeast and some animals. C1 Univ Nebraska, Dept Biochem, Lincoln, NE 68588 USA. NCI, Sect Mol Biol Selenium, Basic Res Lab, NIH, Bethesda, MD 20892 USA. RP Gladyshev, VN (reprint author), Univ Nebraska, Dept Biochem, Lincoln, NE 68588 USA. RI Kryukov, Gregory/A-9592-2008; Gladyshev, Vadim/A-9894-2013; OI Novoselov, Sergey/0000-0003-0104-6492 FU NIGMS NIH HHS [R01 GM061603, GM61603] NR 36 TC 158 Z9 171 U1 3 U2 14 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD JUL 15 PY 2002 VL 21 IS 14 BP 3681 EP 3693 DI 10.1093/emboj/cdf372 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 572PT UT WOS:000176784700013 PM 12110581 ER PT J AU Yu, L Hebert, MC Zhang, YE AF Yu, L Hebert, MC Zhang, YE TI TGF-beta receptor-activated p38 MAP kinase mediates smad-independent TGF-beta responses SO EMBO JOURNAL LA English DT Article DE apoptosis; epithelial-to-mesenchymal transition; p38 MAP kinase; Smad; TGF-beta receptor ID GROWTH-FACTOR-BETA; CELL-CYCLE ARREST; EPITHELIAL-CELLS; SIGNAL-TRANSDUCTION; MESENCHYMAL TRANSDIFFERENTIATION; INHIBITOR P15(INK4B); TRANSCRIPTION FACTOR; I RECEPTORS; APOPTOSIS; EXPRESSION AB Through the action of its membrane-bound type I receptors, transforming growth factor-beta (TGF-beta) elicits a wide range of cellular responses that regulate cell proliferation, differentiation and apoptosis. Many of the signaling responses induced by TGF-beta are mediated by Smad proteins, but certain evidence has suggested that TGF-beta can also signal independently of Smads. We found in mouse mammary epithelial (NMuMG) cells, which respond to TGF-beta treatment in multiple ways, that TGF-beta-induced activation of p38 MAP kinase is required for TGF-beta-induced apoptosis, epithelial-to-mesenchymal transition (EMT), but not growth arrest. We further demonstrated that activation of p38 is independent of Smads using a mutant type I receptor, which is incapable of activating Smads but still retains the kinase activity. This mutant receptor is sufficient to activate p38 and cause NMuMG cells to undergo apoptosis. However, it is not sufficient to induce EMT. These results indicate that TGF-beta receptor signals through multiple intracellular pathways and provide first-hand biochemical evidence for the existence of Smad-independent TGF-beta receptor signaling. C1 NCI, Ctr Canc Res, Cellular & Mol Biol Lab, Bethesda, MD 20892 USA. RP Zhang, YE (reprint author), NCI, Ctr Canc Res, Cellular & Mol Biol Lab, Bethesda, MD 20892 USA. RI Zhang, Ying/G-3657-2015 OI Zhang, Ying/0000-0003-2753-7601 FU Intramural NIH HHS [Z01 BC010419-08] NR 58 TC 404 Z9 426 U1 0 U2 9 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD JUL 15 PY 2002 VL 21 IS 14 BP 3749 EP 3759 DI 10.1093/emboj/cdf366 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 572PT UT WOS:000176784700019 PM 12110587 ER PT J AU Cardozo-Pelaez, F Stedeford, TJ Brooks, PJ Song, SJ Sanchez-Raoms, JR AF Cardozo-Pelaez, F Stedeford, TJ Brooks, PJ Song, SJ Sanchez-Raoms, JR TI Effects of diethylmaleate on DNA damage and repair in the mouse brain SO FREE RADICAL BIOLOGY AND MEDICINE LA English DT Article DE DNA damage; DNA repair; 8-oxoguanine DNA glycosylase; free radicals ID BASE-EXCISION-REPAIR; ALZHEIMERS-DISEASE; OXIDATIVE DAMAGE; POSTISCHEMIC REPERFUSION; GLUTATHIONE DEPLETORS; PARKINSONS-DISEASE; AP-ENDONUCLEASE; CELL LINES; OGG1 GENE; 8-HYDROXY-2'-DEOXYGUANOSINE AB The enzyme 8-oxoguanine DNA glycosylase 1 participates in the repair of damaged DNA by excising the oxidized base 8-hydroxy-2'-deoxyguanosine. We have previously demonstrated that enzymatic activity of this enzyme is inversely related to the levels of the damaged base in specific brain regions. We now report that the activity of 8-oxoguanine DNA glycosylase 1 is increased in a region-specific manner following treatment with diethylmaleate, a compound that reduces glutathione levels in the cell. A single treatment with diethylmaleate elicited a significant increase (similar to2-fold) in the activity of 8-oxoguanine DNA glycosylase I in three brain regions with low basal levels of activity (cerebellum, cortex, and pons/medulla). There was no change in the activity of 8-oxoguanine DNA glycosylase 1 in those regions with high basal levels of activity (hippocampus, caudate/putamen, and midbrain). This is the first report to demonstrate that DNA repair capacity can be upregulated in the CNS, and the increased repair activity correlates with a reduction in the levels of DNA damage. The brain region-specific capacity to deal with increased oxidative damage to DNA may be responsible, in part, for the vulnerability of specific neuronal populations with aging, sources of oxidative stress, and neurodegenerative diseases. (C) 2002 Elsevier Science Inc. C1 Univ S Florida, Dept Neurol MDC 55, Ctr Aging & Brain Repair, Tampa, FL 33612 USA. NIAAA, Neurogenet Lab, NIH, Rockville, MD USA. James A Haley Vet Hosp, Res Serv, Tampa, FL 33612 USA. RP Sanchez-Raoms, JR (reprint author), Univ S Florida, Dept Neurol MDC 55, Ctr Aging & Brain Repair, 12901 Bruce B Downs Blvd, Tampa, FL 33612 USA. RI Sanchez-Ramos, Juan/A-1188-2009 OI Sanchez-Ramos, Juan/0000-0002-3391-7857 NR 31 TC 18 Z9 18 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0891-5849 J9 FREE RADICAL BIO MED JI Free Radic. Biol. Med. PD JUL 15 PY 2002 VL 33 IS 2 BP 292 EP 298 AR PII S0891-5849(02)00881-X DI 10.1016/S0891-5849(02)00881-X PG 7 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 574RY UT WOS:000176904400015 PM 12106825 ER PT J AU Cui, CY Durmowicz, M Tanaka, TS Hartung, AJ Tezuka, T Hashimoto, K Ko, MSH Srivastava, AK Schlessinger, D AF Cui, CY Durmowicz, M Tanaka, TS Hartung, AJ Tezuka, T Hashimoto, K Ko, MSH Srivastava, AK Schlessinger, D TI EDA targets revealed by skin gene expression profiles of wild-type, Tabby and Tabby EDA-A1 transgenic mice SO HUMAN MOLECULAR GENETICS LA English DT Article ID HYPOHIDROTIC ECTODERMAL DYSPLASIA; EPIDERMAL GROWTH-FACTOR; HAIR FOLLICLE MORPHOGENESIS; C-FOS; ECTODYSPLASIN-A; KAPPA-B; BETA-CATENIN; PROTEIN; RECEPTOR; MUTATIONS AB Mutations in the EDA gene cause anhidrotic ectodermal dysplasia (EDA), with lesions in skin appendage formation. To begin to analyze EDA pathways, we have used expression profiling on 15000-gene mouse cDNA microarrays, comparing adult mouse skin from wild-type, EDA-defective (Tabby) mice, and Tabby mice supplemented with the EDA-A1 isoform, which is sufficient to rescue multiple Tabby phenotypes. Given the sensitivity of the current microarray system, 8500 genes (60%) were estimated to be expressed, including transcription factors and growth-regulatory genes that had not previously been identified in skin; but only 24 (0.16%), one-third of them novel, showed significant differences between wild type and Tabby. An additional eight genes not included in the 15000 gene set were shown to have expression differences by real-time RT-PCR. Sixteen of 32 affected genes were restored significantly toward wild-type levels in EDA-A1 transgenic Tabby mice. Significant up-regulation in Tabby skin was observed for several dermal matrix genes, including Col1a1, Col1a2, Col3a1 and Sparc. In contrast, down-regulation occurred for the NEMO/ NF-kB pathway, already implicated in skin appendage formation, and even more markedly for a second pathway, JNK/c-jun/c-fos and their target genes, that has not previously been clearly associated with skin development. These data are consistent with the regulation of the NF-kB pathway by EDA, and support its involvement in the regulation of the JNK pathway as well. C1 NIA, Genet Lab, NIH, Baltimore, MD 21224 USA. Greenwood Genet Ctr, JC Self Res Inst Human Genet, Greenwood, SC 29646 USA. Kinki Univ, Sch Med, Dept Dermatol, Osaka 589, Japan. Wayne State Univ, Sch Med, Dept Dermatol & Syphilol, Detroit, MI 48201 USA. RP Schlessinger, D (reprint author), NIA, Genet Lab, NIH, Baltimore, MD 21224 USA. RI Ko, Minoru/B-7969-2009 OI Ko, Minoru/0000-0002-3530-3015 NR 55 TC 29 Z9 32 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 J9 HUM MOL GENET JI Hum. Mol. Genet. PD JUL 15 PY 2002 VL 11 IS 15 BP 1763 EP 1773 DI 10.1093/hmg/11.15.1763 PG 11 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 575VU UT WOS:000176969800008 PM 12095918 ER PT J AU Piccirillo, CA Letterio, JJ Thornton, AM McHugh, RS Mamura, M Mizuhara, H Shevach, EM AF Piccirillo, CA Letterio, JJ Thornton, AM McHugh, RS Mamura, M Mizuhara, H Shevach, EM TI CD4(+)CD25(+) regulatory T cells can mediate suppressor function in the absence of transforming growth factor beta 1 production and responsiveness SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE CD4(+) suppressor T cells; autoimmunity; tolerance; immunoregulation; IL-2 receptor ID ORGAN-SPECIFIC AUTOIMMUNITY; IMMUNOLOGICAL SELF-TOLERANCE; TGF-BETA; IN-VITRO; INTESTINAL INFLAMMATION; IMMUNOREGULATORY CELLS; TARGETED DISRUPTION; EFFECTOR FUNCTION; PERIPHERAL-BLOOD; MICE AB CD4(+)CD25(+) regulatory T cells inhibit organ-specific autoimmune diseases induced by CD4(+)CD25(-) T cells and are potent suppressors of T cell activation in vitro. Their mechanism of suppression remains unknown. but most in vitro studies suggest that it is cell contact-dependent and cytokine independent. The role of TGF-beta1 in CD4(+)CD25(+) suppressor function remains unclear. While most studies have failed to reverse suppression with anti-transforming growth factor (TGF)-beta1 in vitro, one recent study has reported that CD4(+)CD25(+) T cells express cell surface TGF-beta1 and that suppression can be completely abrogated by high concentrations of anti-TGF-beta suggesting that cell-associated TGF-beta1 was the primary effector of CD4(+)CD25(+)-mediated suppression. Here, we have reevaluated the role of TGF-beta1 in CD4(+)CD25(+)-mediated suppression, Neutralization of TGF-beta1 with either monoclonal antibody (mAb) or soluble TGF-betaRII-Fc did not reverse in vitro suppression mediated by resting or activated CD4(+)CD25(+) T cells. Responder T cells from Smad3(-/-) or dominant-negative TGF-beta type RII transgenic (DNRIITg) mice, that are both unresponsive to TGF-beta1-induced growth arrest, were as susceptible to CD4(+)CD25(+)-mediated suppression as T cells from wildtype mice, Furthermore, CD4(+)CD25(+) T cells from neonatal TGF-beta1(-/-) mice were as suppressive as CD4(+)CD25(+) from TGF-beta1(+/+) mice. Collectively, these results demonstrate that CD4(+)CD25(+) suppressor function can occur independently of TGF-beta. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Shevach, EM (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Rm 11N315,10 Ctr Dr, Bethesda, MD 20892 USA. OI Mamura, Mizuko/0000-0003-4531-0144 NR 47 TC 447 Z9 484 U1 0 U2 7 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD JUL 15 PY 2002 VL 196 IS 2 BP 237 EP 245 DI 10.1084/jem.20020590 PG 9 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 576PG UT WOS:000177013200009 PM 12119348 ER PT J AU Ishii, KJ Takeshita, F Gursel, I Gursel, M Conover, J Nussenzweig, A Klinman, DM AF Ishii, KJ Takeshita, F Gursel, I Gursel, M Conover, J Nussenzweig, A Klinman, DM TI Potential role of phosphatidylinositol 3 kinase, rather than DNA-dependent protein kinase, in CpG DNA-induced immune activation SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE toll-like receptors; wortmannin; dendritic cells; innate immunity; cytokines ID BACTERIAL-DNA; V(D)J RECOMBINATION; IMMUNOSTIMULATORY DNA; CELLS; MATURATION; RECOGNITION; 3-KINASES; INNATE; MOTIFS; PKCS AB Unmethylated CpG motifs present in bacterial DNA stimulate a strong innate immune response. There is evidence that DNA-dependent protein kinase (DNA-PK) mediates CpG signaling. Specifically, wortmannin (an inhibitor of phosphatidylinositol 3 kinase [PI3]-kinases including DNA-PK) interferes with CpG-dependent cell activation, and DNA-PK knockout (KO) mice fail to respond to CpG stimulation. Current studies establish that wortmannin actually inhibits the uptake and colocalization of CpG DNA with toll-like receptor (TLR)-9 in endocytic vesicles, thereby preventing CpG-induced activation of the NF-kappaB signaling cascade. We find that DNA-PK is not involved in this process, since three strains of DNA-PK KO mice responded normally to CpG DNA. These results support a model in which CpG signaling is mediated through TLR-9 but not DNA-PK, and suggest that wortmannin-sensitive member(s) of the PI3-kinase family play a critical role in shuttling CpG DNA to TLR-9. C1 US FDA, Ctr Biol Evaluat & Res, Div Viral Prod, Sect Retroviral Immunol, Bethesda, MD 20892 USA. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Klinman, DM (reprint author), US FDA, Ctr Biol Evaluat & Res, Div Viral Prod, Sect Retroviral Immunol, Bldg 29A,Rm 3D10, Bethesda, MD 20892 USA. RI Gursel, Mayda /H-1812-2012; Ishii, Ken/B-1685-2012; OI Ishii, Ken/0000-0002-6728-3872; Gursel, Ihsan/0000-0003-3761-1166; Gursel, Mayda/0000-0003-0044-9054 NR 26 TC 108 Z9 112 U1 1 U2 5 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD JUL 15 PY 2002 VL 196 IS 2 BP 269 EP 274 DI 10.1084/jem.20020773 PG 6 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 576PG UT WOS:000177013200013 PM 12119352 ER PT J AU Ishaq, M Fan, M Wigmore, K Gaddam, A Natarajan, V AF Ishaq, M Fan, M Wigmore, K Gaddam, A Natarajan, V TI Regulation of retinoid X receptor responsive element-dependent transcription in T lymphocytes by Ser/Thr phosphatases: Functional divergence of protein kinase C (PKC)theta and PKC alpha in mediating calcineurin-induced transactivation SO JOURNAL OF IMMUNOLOGY LA English DT Article ID IL-2 PROMOTER; CELL-CYCLE; RXR-ALPHA; ACTIVATION; THETA; COMPLEX; PHOSPHORYLATION; COSTIMULATION; ASSOCIATION; EXPRESSION AB T lymphocyte activation signals regulate the expression and transactivation function of retinoid X receptor (RXR) a through an interplay of complex signaling cascades that are not yet fully understood. We show that cellular Ser/Thr protein phosphatases (PPs) play an important role in mediating these processes. Inhibitors specific for PP1 and PP2A decreased basal expression of RXRalpha RNA and protein in T lymphocyte leukemia Jurkat cells and prevented activation-induced RXRalpha accumulation in these cells. In addition, these inhibitors attenuated the RXR responsive element (RXRE)-dependent transcriptional activation in transient transfection assays. Inhibitors of calcineurin (CN), by contrast, did not have any effect on the basal RXRalpha expression and even augmented activation-induced RXRalpha expression. Expression of a dominant-active (DA) mutant of CN together with a DA mutant of protein kinase C (PKQ)theta, a novel PKC isoform, significantly increased RXRE-dependent transcription. Expression of catalytically inactive PKCtheta or a dominant-negative mutant of PKCtheta failed to synergize with CN and did not increase RXRE-dependent transcription. Expression of a DA mutant of PKCalpha or treatment with PMA was found to attenuate PKCtheta and CN synergism. We conclude that PP1, MA, and CN regulate levels and transcriptional activation function of RXRa in T cells. In addition, CN synergizes with PKCtheta to induce RXRE-dependent activation, a cooperative function that is antagonized by the activation of the conventional PKCalpha isoform. Thus, PKCtheta and PKCalpha may function as positive and negative modulators, respectively, of CN-regulated RXRE-dependent transcription during T cell activation. C1 NCI, Mol Cell Biol Lab, Sci Applicat Int Corp Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Ishaq, M (reprint author), NCI, Mol Cell Biol Lab, Sci Applicat Int Corp Frederick, Frederick Canc Res & Dev Ctr, Bldg 550,Room 104, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 39 TC 14 Z9 15 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD JUL 15 PY 2002 VL 169 IS 2 BP 732 EP 738 PG 7 WC Immunology SC Immunology GA 572BK UT WOS:000176753500014 PM 12097375 ER PT J AU Cunningham, CK Chaix, ML Rekacewicz, C Britto, P Rouzioux, C Gelber, RD Dorenbaum, A Delfraissy, JF Bazin, B Mofenson, L Sullivan, JL AF Cunningham, CK Chaix, ML Rekacewicz, C Britto, P Rouzioux, C Gelber, RD Dorenbaum, A Delfraissy, JF Bazin, B Mofenson, L Sullivan, JL CA Pediat AIDS Clin Trials Grp Protoc TI Development of resistance mutations in women receiving standard antiretroviral therapy who received intrapartum nevirapine to prevent perinatal human immunodeficiency virus type 1 transmission: A substudy of Pediatric AIDS Clinical Trials Group protocol 316 SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 8th Conference on Retroviruses and Opportunistic Infections CY FEB 04-08, 2001 CL CHICAGO, ILLINOIS ID ZIDOVUDINE; HIV-1; PREVALENCE; INFECTION AB Pediatric AIDS Clinical Trials Group protocol 316 was an international, multicenter, placebo-controlled trial comparing single-dose oral nevirapine (200 mg to mother and 2 mg/kg to infant) with placebo in human immunodeficiency virus (HIV)-infected pregnant women receiving standard antiretroviral therapy. This substudy evaluated the emergence of nevirapine-resistance mutations at 6 weeks postpartum in a subgroup of participants. Maternal risk factors for the emergence of nevirapine-resistance mutations were evaluated. Mutations associated with nevirapine resistance were detectable at delivery, prior to receipt of study drug, in 5 (2.3%) of 217 women. Fourteen (15%; 95% confidence interval, 8%-23%) of 95 women who received intrapartum nevirapine developed a nevirapine-resistance mutation 6 weeks postpartum. The most common mutation was K103N, which was present in 10 women. The risk for development of a new nevirapine-resistance mutation did not correlate with CD4 cell count or HIV-1 RNA load at delivery or with type of antepartum antiretroviral therapy. The risk of nevirapine resistance should be considered when determining the risks or benefits of intrapartum nevirapine in women receiving antepartum antiretroviral therapy. C1 SUNY Upstate Med Univ, Dept Pediat, Syracuse, NY 13210 USA. Harvard Univ, Sch Publ Hlth, Stat & Data Anal Ctr, Boston, MA 02115 USA. Univ Massachusetts, Sch Med, Dept Pediat, Worcester, MA USA. Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94143 USA. NICHHD, NIH, Rockville, MD USA. Hop Necker Enfants Malad, Dept Virol, Paris, France. SIDA, Agence Natl Rech, Paris, France. INSERM, Serv Commun 10, Villejuif, France. Med Interne, Le Kremlin Bicetre, France. RP Cunningham, CK (reprint author), SUNY Upstate Med Univ, Dept Pediat, 750 E Adams St, Syracuse, NY 13210 USA. FU NIAID NIH HHS [AI32907, AI41110, AI42845] NR 17 TC 81 Z9 84 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUL 15 PY 2002 VL 186 IS 2 BP 181 EP 188 DI 10.1086/341300 PG 8 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 570QG UT WOS:000176668500005 PM 12134253 ER PT J AU Biggar, RJ Janes, M Pilon, R Roy, R Broadhead, R Kumwenda, N Taha, TET Cassol, S AF Biggar, RJ Janes, M Pilon, R Roy, R Broadhead, R Kumwenda, N Taha, TET Cassol, S TI Human immunodeficiency virus type 1 infection in twin pairs infected at birth SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID TO-CHILD TRANSMISSION; AFRICAN INFANTS; HIV-1; MACROPHAGES; MONOCYTES; VARIANTS AB Host genetic factors may influence the course of human immunodeficiency virus (HIV) infection. In Blantyre, Malawi, polymerase chain reaction was used to identify twin pairs who were concordantly HIV-1-infected in utero or perinatally and then to examine strain divergence or virus levels in identical and fraternal twin pairs. Among 315 twin pairs, both infants in 14 fraternal and 5 identical pairs were found to be infected at the same visit. Among 10 pairs, HIV-1 sequences were determined for both infants at greater than or equal to1 time point. HIV levels had a common profile in both fraternal and identical twin pairs. Identical twins were not always infected by the same quasi species, indicating that their mothers had multiple quasi species capable of infecting their infants. Subsequent viral divergence appears to depend on quasi-species stability rather than on genetically controlled host immune responses. Thus, given infection, factors intrinsic to HIV-1 are more important than host genetics in viral evolution. C1 NCI, Viral Epidemiol Branch, Rockville, MD 20852 USA. Johns Hopkins Sch Publ Hlth, Baltimore, MD USA. Univ Ottawa, Ottawa, ON, Canada. St Louis Cty Police Crime Lab, St Louis, MO USA. Univ Malawi, Dept Paediat, Zomba, Malawi. Johns Hopkins Univ Univ Malawi Project, Blantyre, Malawi. Univ Natal, Nelson R Mandela Sch Med, ZA-4001 Durban, South Africa. Univ Natal, Africa Ctr, ZA-4001 Durban, South Africa. RP Biggar, RJ (reprint author), NCI, Viral Epidemiol Branch, 6120 Execut Blvd, Rockville, MD 20852 USA. NR 13 TC 9 Z9 9 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUL 15 PY 2002 VL 186 IS 2 BP 281 EP 285 DI 10.1086/341453 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 570QG UT WOS:000176668500019 PM 12134267 ER PT J AU Kim, OJ Ariano, MA Lazzarini, RA Levine, MS Sibley, DR AF Kim, OJ Ariano, MA Lazzarini, RA Levine, MS Sibley, DR TI Neurofilament-m interacts with the D-1 dopamine receptor to regulate cell surface expression and desensitization SO JOURNAL OF NEUROSCIENCE LA English DT Article DE D-1 receptor; neurofilament-M; yeast two-hybrid; interaction; desensitization; colocalization ID NEURONAL INTERMEDIATE FILAMENTS; ANTIRECEPTOR ANTISERA; RAT-BRAIN; PROTEIN; LOCALIZATION; PHOSPHORYLATION; ANTIBODIES; TRANSPORT; SYNAPSES; SYSTEM AB We used the yeast two-hybrid assay to identify novel proteins that interact with the D-1 dopamine receptor. The third cytoplasmic loop (residues 217-273) of the rat D-1 receptor was used as bait to identify clones encoding interacting proteins from a rat brain cDNA library. This identified two clones encoding the C terminus of rat neurofilament-M (NF-M) (residues 782-846). The NF-M clone did not interact with the third cytoplasmic loops of the rat D-2, D-3, or D-4 receptors, but showed weak interaction with that of the D-5 receptor. Coexpression of full-length NF-M with the D-1 receptor in HEK-293 cells resulted in >50% reduction of receptor binding accompanied by a reduction in D-1 receptor-mediated cAMP accumulation. NF-M had no effect on the expression of other dopamine receptor subtypes. Using a D-1 receptor-green fluorescent protein chimera and confocal fluorescence microscopy, we found that NF-M reduced D-1 receptor expression at the cell surface and promoted accumulation of the receptor in the cytosol. Interestingly, the D-1 receptors that were expressed at the cell surface in the presence of NF-M were resistant to agonist-induced desensitization. Cellular colocalization of NF-M and the D-1 receptor in the rat brain was examined by epifluorescence microscopy. These experiments showed that similar to50% of medium-sized striatal neurons expressed both proteins. Colocalization was also observed in pyramidal cells and interneurons within the frontal cortex. Similar immunohistochemical analyses using NF-M-deficient mice showed decrements in D-1 receptor expression compared with control mice. These results suggest that NF-M interacts with the D-1 receptor in vivo and may modify its expression and regulation. C1 NINDS, Mol Neuropharmacol Sect, NIH, Bethesda, MD 20892 USA. Finch Univ Hlth Sci Chicago Med Sch, Dept Neurosci, N Chicago, IL 60064 USA. CUNY Mt Sinai Sch Med, Dept Biochem & Mol Biol, New York, NY 10029 USA. Univ Calif Los Angeles, Mental Retardat Res Ctr, Los Angeles, CA 90095 USA. RP Sibley, DR (reprint author), NINDS, Mol Neuropharmacol Sect, NIH, Bldg 10,Room 5C108,10 Ctr Dr,MSC 1406, Bethesda, MD 20892 USA. FU NINDS NIH HHS [NS33538] NR 32 TC 55 Z9 56 U1 0 U2 0 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD JUL 15 PY 2002 VL 22 IS 14 BP 5920 EP 5930 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 573PR UT WOS:000176840500017 PM 12122054 ER PT J AU Ye, P Li, LQ Richards, RG DiAugustine, RP D'Ercole, AJ AF Ye, P Li, LQ Richards, RG DiAugustine, RP D'Ercole, AJ TI Myelination is altered in insulin-like growth factor-I null mutant mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE IGF-I; IGF-II; oligodendrocytes; oligodendrocyte precursors; myelination; development ID GLUTATHIONE-S-TRANSFERASE; CENTRAL-NERVOUS-SYSTEM; IGF BINDING PROTEIN-1; TRANSGENIC MICE; OLIGODENDROCYTE DEVELOPMENT; PROTEOLIPID PROTEIN; RAT-BRAIN; DIFFERENTIAL EXPRESSION; PROGENITOR CELLS; GENE-EXPRESSION AB Increasing evidence indicates that insulin-like growth factor-I (IGF-I) has an important role in oligodendrocyte development. In this study, we examined myelination during postnatal development in IGF-I knock-out (KO) mice by assessing myelin staining, the expression of myelin basic protein (MBP) and proteolipid protein (PLP), two major myelin-specific proteins, and the number of oligodendrocytes and their precursors. For comparison, we also measured the expression of median subunit of the neuron-specific intermediate filament, M-neurofilament (MNF), to obtain an index of the effects of IGF-I deficiency on neurons. We found that myelin staining, MBP and PLP expression, and the percentage of oligodendrocytes and their precursors are significantly reduced in all brain regions of developing IGF-I KO mice but are similar to controls in adult IGF-I KO mice. In contrast, the abundance of M-NF was decreased in both the developing and adult brain of IGF-IKO mice. We also found that IGF-II protein abundance is increased in the brains of IGF-IKO mice. Our data indicate, therefore, that myelination during early development is altered in the absence of IGF-I by mechanisms that involve a reduction in oligodendrocyte proliferation and development. Although neuronal actions cannot be excluded in the myelin normalization, the reduced axonal growth suggested by the reduced M-NF expression makes a role for neuronal factors less compelling. These data suggest that IGF-I plays a significant role in myelination during normal early development and that IGF-II can compensate in part for IGF-I actions on myelination. C1 Univ N Carolina, Dept Pediat, Chapel Hill, NC 27599 USA. NIEHS, Hormones & Canc Grp, Res Triangle Pk, NC 27709 USA. RP D'Ercole, AJ (reprint author), Univ N Carolina, Dept Pediat, CB 7220, Chapel Hill, NC 27599 USA. RI Ye, Ping/A-7144-2009; Messier, Claude/A-2322-2008 OI Messier, Claude/0000-0002-4791-1763 FU NICHD NIH HHS [HD08299]; NINDS NIH HHS [NS38891] NR 52 TC 134 Z9 139 U1 0 U2 2 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD JUL 15 PY 2002 VL 22 IS 14 BP 6041 EP 6051 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 573PR UT WOS:000176840500028 PM 12122065 ER PT J AU Fallon, JR Rafii, M Hagiwara, H McKechnie, BA McQuillan, DM Young, MA AF Fallon, JR Rafii, M Hagiwara, H McKechnie, BA McQuillan, DM Young, MA TI Biglycan is an extracellular scaffolding molecular for the dystrophin-associated protein complex that constitutes a novel link to the extracellular matrix SO JOURNAL OF THE NEUROLOGICAL SCIENCES LA English DT Meeting Abstract CT 10th International Congress on Neuromuscular Diseases CY JUL 07-12, 2002 CL VANCOUVER, CANADA C1 Brown Univ, Dept Neurosci, Providence, RI 02912 USA. Lifecell Corp, Branchburg, NJ USA. NIDR, NIH, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-510X J9 J NEUROL SCI JI J. Neurol. Sci. PD JUL 15 PY 2002 VL 199 SU 1 MA 09 BP S3 EP S3 PG 1 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 632WT UT WOS:000180247400010 ER PT J AU Frith, MC Spouge, JL Hansen, U Weng, ZP AF Frith, MC Spouge, JL Hansen, U Weng, ZP TI Statistical significance of clusters of motifs represented by position specific scoring matrices in nucleotide sequences SO NUCLEIC ACIDS RESEARCH LA English DT Article ID TRANSCRIPTION FACTOR CP2; SERUM RESPONSE FACTOR; ALPHA-ACTIN PROMOTER; DNA-BINDING SITES; GENE-EXPRESSION; HIV-1 TRANSCRIPTION; REGULATORY ELEMENTS; MESSENGER-RNA; MOUSE GENOME; IN-VITRO AB The human genome encodes the transcriptional control of its genes in clusters of cis-elements that constitute enhancers, silencers and promoter signals. The sequence motifs of individual cis-elements are usually too short and degenerate for confident detection. In most cases, the requirements for organization of cis-elements within these clusters are poorly understood. Therefore, we have developed a general method to detect local concentrations of cis-element motifs, using predetermined matrix representations of the cis-elements, and calculate the statistical significance of these motif clusters. The statistical significance calculation is highly accurate not only for idealized, pseudo-random DNA, but also for real human DNA. We use our method 'cluster of motifs E-value tool' (COMET) to make novel predictions concerning the regulation of genes by transcription factors associated with muscle. COMET performs comparably with two alternative state-of-the-art techniques, which are more complex and lack E-value calculations. Our statistical method enables us to clarify the major bottleneck in the hard problem of detecting cis-regulatory regions, which is that many known enhancers do not contain very significant clusters of the motif types that we search for. Thus, discovery of additional signals that belong to these regulatory regions will be the key to future progress. C1 Boston Univ, Bioinformat Program, Boston, MA 02215 USA. Boston Univ, Dept Biomed Engn, Boston, MA 02215 USA. Boston Univ, Dept Biol, Boston, MA 02215 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. RP Weng, ZP (reprint author), Boston Univ, Bioinformat Program, 44 Cummington St, Boston, MA 02215 USA. EM zhiping@bu.edu FU NCI NIH HHS [CA81157, R01 CA081157, R01 CA081157-05] NR 71 TC 85 Z9 92 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD JUL 15 PY 2002 VL 30 IS 14 BP 3214 EP 3224 DI 10.1093/nar/gkf438 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 579AY UT WOS:000177154300031 PM 12136103 ER PT J AU Lavrik, OI Kolpashchikov, DM Prasad, R Sobol, RW Wilson, SH AF Lavrik, OI Kolpashchikov, DM Prasad, R Sobol, RW Wilson, SH TI Binary system for selective photoaffinity labeling of base excision repair DNA polymerases SO NUCLEIC ACIDS RESEARCH LA English DT Article ID REPLICATION PROTEIN-A; BETA; CHROMOSOMES; SUBSTRATE; LINKING; ANALOGS AB A system of photoaffinity reagents for selective labeling of DNA polymerases In extracts has been examined. To create the photoreactive DNA probe in situ, DNA substrates containing a synthetic abasic site are Incubated in mouse embryonic fibroblast (MEF) cellular extract In the presence of base-substituted arylazido derivatives of dNTPs. This results In synthesis of a photoreactive long patch base excision repair (BER) intermediate. The arylazido photoreactive group Is then activated through energy transfer from the pyrene group of a dNTP analog (Pyr-dUTP), following 365 nm UV light exposure. Pyr-dUTP binds to the active site of DNA polymerases, and the pyrene group, when excited by 365 nm UV light, activates the nearby photoreactive group In the BER Intermediate resulting In crosslinking of DNA-bound DNA polymerases. Under these conditions, various DNA binding proteins that are unable to bind Pyr-dUTP are not crosslinked to DNA. DNA polymerase beta Is the predominant crosslinked protein observed in the MEF extract. In contrast, several other DNA binding proteins are labeled under conditions of direct UV light activation of the photoreactive group at 312 nm. This study illustrates use of a new method of selective labeling of DNA polymerases In a crude cellular extract. C1 NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. Russian Acad Sci, Siberian Div, Novosibirsk Bioorgan Chem Inst, Novosibirsk 630090, Russia. RP Wilson, SH (reprint author), NIEHS, Struct Biol Lab, NIH, 111 T W Alexander Dr, Res Triangle Pk, NC 27709 USA. RI Lavrik, Olga /G-4641-2013; Sobol, Robert/E-4125-2013 OI Sobol, Robert/0000-0001-7385-3563 NR 26 TC 14 Z9 14 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD JUL 15 PY 2002 VL 30 IS 14 AR e73 DI 10.1093/nar/gnf073 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 579AY UT WOS:000177154300010 PM 12136121 ER PT J AU Charan, RD McKee, TC Gustafson, KR Pannell, LK Boyd, MR AF Charan, RD McKee, TC Gustafson, KR Pannell, LK Boyd, MR TI Thorectandramine, a novel beta-carboline alkaloid from the marine sponge Thorectandra sp. SO TETRAHEDRON LETTERS LA English DT Article DE Thorectandra; marine natural product; beta-carboline alkaloid; sponge AB The isolation, structure elucidation and biological activity of a novel hexacyclic quaternary alkaloid, thorectandramine (1), from a Palauan sponge of the genus Thorectandra are reported. (C) 2002 Published by Elsevier Science Ltd. C1 NCI, Canc Res Ctr, Mol Targets Drug Discovery Program, Frederick, MD 21702 USA. NIDDK, Bioorgan Chem Lab, Bethesda, MD 20892 USA. RP McKee, TC (reprint author), NCI, Canc Res Ctr, Mol Targets Drug Discovery Program, Frederick, MD 21702 USA. NR 10 TC 29 Z9 29 U1 0 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0040-4039 J9 TETRAHEDRON LETT JI Tetrahedron Lett. PD JUL 15 PY 2002 VL 43 IS 29 BP 5201 EP 5204 AR PII S0040-4039(02)01023-7 DI 10.1016/S0040-4039(02)01023-7 PG 4 WC Chemistry, Organic SC Chemistry GA 570BX UT WOS:000176639100048 ER PT J AU Williams, L Chou, MW Yan, J Young, JF Chan, PC Doerge, DR AF Williams, L Chou, MW Yan, J Young, JF Chan, PC Doerge, DR TI Toxicokinetics of riddelliine, a carcinogenic pyrrolizidine alkaloid, and metabolites in rats and mice SO TOXICOLOGY AND APPLIED PHARMACOLOGY LA English DT Article DE riddelliine; pyrrolizidine alkaloid; mass spectrometry; toxicokinetics ID IN-VIVO AB Riddelliine is a representative pyrrolizidine alkaloid, a class of naturally occurring toxic phytochemicals present in plant species worldwide. Human exposure to pyrrolizidine alkaloids occurs through consumption of herbal dietary supplements, including comfrey, and through contaminated livestock products (e.g., milk). A recently completed 2-year bioassay of riddelliine carcinogenicity showed that male and female rats and male mice, but not female mice, developed liver tumors. The toxicokinetics of riddelliine and two metabolites, the N-oxide and retronecine, were determined in serum following an oral gavage dose in male and female rats and mice using a validated liquid chromatography-electrospray mass spectrometric method. The results are consistent with extensive metabolism of riddelliine and its more polar metabolites prior to excretion. It is concluded that factors other than toxicokinetics are responsible for the observed species/sex specificity of gross toxicity or liver tumor induction in rats and mice. C1 Natl Ctr Toxicol Res, Jefferson, AR 72079 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Doerge, DR (reprint author), Natl Ctr Toxicol Res, Jefferson, AR 72079 USA. NR 13 TC 18 Z9 20 U1 0 U2 3 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0041-008X J9 TOXICOL APPL PHARM JI Toxicol. Appl. Pharmacol. PD JUL 15 PY 2002 VL 182 IS 2 BP 98 EP 104 DI 10.1006/taap.2002.9441 PG 7 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 580XC UT WOS:000177261200002 PM 12140173 ER PT J AU Petricoin, EF Mills, GB Kohn, EC Liotta, LA AF Petricoin, EF Mills, GB Kohn, EC Liotta, LA TI Proteomic patterns in serum and identification of ovarian cancer - Reply SO LANCET LA English DT Letter C1 US FDA, NIH, Clin Proteom Program, Dept Therapeut Prot,Ctr Biol Evaluat & Res, Bethesda, MD 20892 USA. Univ Texas, MD Anderson Canc Ctr, Dept Med Therapeut, Div Canc Med, Houston, TX 77030 USA. NCI, Pathol Lab, Ctr Canc Res, Bethesda, MD 20892 USA. RP Petricoin, EF (reprint author), US FDA, NIH, Clin Proteom Program, Dept Therapeut Prot,Ctr Biol Evaluat & Res, Bldg 29A,Room 2B02,880 Rockville Pike, Bethesda, MD 20892 USA. NR 5 TC 30 Z9 31 U1 0 U2 3 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUL 13 PY 2002 VL 360 IS 9327 BP 170 EP 171 DI 10.1016/S0140-6736(02)09391-1 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 573PB UT WOS:000176839100034 ER PT J AU Remaley, AT Bark, S Walts, AD Freeman, L Shulenin, S Annilo, T Elgin, E Rhodes, HE Joyce, C Dean, M Santamarina-Fojo, S Brewer, HB AF Remaley, AT Bark, S Walts, AD Freeman, L Shulenin, S Annilo, T Elgin, E Rhodes, HE Joyce, C Dean, M Santamarina-Fojo, S Brewer, HB TI Comparative genome analysis of potential regulatory elements in the ABCG5-ABCG8 gene cluster SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE sitosterolemia; phytosterols; ABCG5; ABCG8; ABC transporter ID DIETARY-CHOLESTEROL ABSORPTION; BIDIRECTIONAL PROMOTER; TRANSCRIPTION FACTORS; SITOSTEROLEMIA; GATA; ABCG5; MUTATIONS; PROTEIN; DOMAIN; LOCUS AB The excretion of sterols from the liver and intestine is regulated by the ABCG5 and ABCG8 transporters. To identify potential regulatory elements, 152kb of the human ABCG5-ABCG8 gene cluster was sequenced and comparative genome analysis was performed. The two genes are oriented in a head-to-head configuration and are separated by a 374-bp intergenic region, which is highly conserved among several species. Using a reporter construct, the intergenic region was found to act as a bidirectional promoter. A conserved GATA site in the intergenic region was shown by site-directed mutagenesis to act Lis a repressor for the ABCG5 promoter. The intergenic region was also shown to be partially responsive to treatment by LXR agonists. In Summary, several potential regulatory elements were found for the ABCG5 and ABCG8 genes. and the intergenic region was found to act as a bidirectional promoter. Published by Elsevier Science (USA). C1 NHLBI, NIH, Bethesda, MD 20892 USA. NCI, Lab Genom Divers, NIH, Frederick, MD USA. Walter Reed Army Med Ctr, Walter Reed Army Med Ctr, Dept Med, Serv Cardiol, Washington, DC USA. RP Remaley, AT (reprint author), NHLBI, NIH, Bldg 10-2C-433,10 Ctr Dr, Bethesda, MD 20892 USA. RI Dean, Michael/G-8172-2012; Annilo, Tarmo/J-2900-2013 OI Dean, Michael/0000-0003-2234-0631; Annilo, Tarmo/0000-0002-9588-3058 NR 25 TC 39 Z9 39 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUL 12 PY 2002 VL 295 IS 2 BP 276 EP 282 AR PII S0006-291X(02)00652-6 DI 10.1016/S0006-291X(02)00652-6 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 574WA UT WOS:000176911500010 PM 12150943 ER PT J AU Gu, HY Wang, SX Messam, CA Yao, ZB AF Gu, HY Wang, SX Messam, CA Yao, ZB TI Distribution of nestin immunoreactivity in the normal adult human forebrain SO BRAIN RESEARCH LA English DT Article DE nestin; adult human; basal forebrain; hippocampus; neuroprogenitor cell ID CENTRAL-NERVOUS-SYSTEM; INTERMEDIATE FILAMENTS; HUMAN CNS; CELLS; EXPRESSION; BRAIN; STEM; BIOLOGY; TISSUE; INJURY AB The distribution of nestin immunoreactivity was studied in the whole normal adult human forebrains using new anti-human nestin mouse monoclonal and rabbit polyclonal antiserum. The nestin immunoreactive cells could be divided into three types according to their morphological characteristics. The first type contained neuron-like nestin immunoreactive cells, distributed in CA1-3 of hippocampus, septum, the nucleus of diagonal band, amygdala and basal nucleus of Meynert. The second type contained astrocyte-like cells, distributed in the subependymal zone and subgranular layer of dentate gyrus. The third type of cells had smaller cell bodies and fewer processes, also distributed in the subependymal zone and subgranular layer of dentate gyrus. Double immunohistochemical staining showed that the nestin positive, neuron-like cells in the nucleus of diagonal band and hippocampus also expressed NSE. However, the astrocyte-like nestin immunoreactive cells of the subependymal zone and subgranular layer of dentate gyrus were not double labeled with GFAP. Although some nestin immunoreactive fibers were distributed in the infundibulum, no nestin-immunoreactive cells were detected in the cortex. These data indicate that nestin exist in the adult human brain outside of the subependymal zone and dentate gyrus and also implies that nestin-immunoreactive cells may play a role in the modulation of basal forebrain function. (C) 2002 Elsevier Science B.V. All rights reserved. C1 Sun Yat Sen Univ Med Sci, Dept Neurobiol & Anat, Guangzhou 510089, Peoples R China. NINDS, Lab Mol Med & Neurosci, NIH, Bethesda, MD 20892 USA. RP Sun Yat Sen Univ Med Sci, Dept Neurobiol & Anat, 74 Zhongshan 2nd Rd, Guangzhou 510089, Peoples R China. EM zbyao@gzsums.edu.cn NR 24 TC 45 Z9 52 U1 1 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 EI 1872-6240 J9 BRAIN RES JI Brain Res. PD JUL 12 PY 2002 VL 943 IS 2 BP 174 EP 180 AR PII S0006-8993(02)02615-X DI 10.1016/S0006-8993(02)02615-X PG 7 WC Neurosciences SC Neurosciences & Neurology GA 580FE UT WOS:000177223800002 PM 12101039 ER PT J AU Makino, S Smith, MA Gold, PW AF Makino, S Smith, MA Gold, PW TI Regulatory role of glucocorticoids and glucocorticoid receptor mRNA levels on tyrosine hydroxylase gene expression in the locus coeruleus during repeated immobilization stress SO BRAIN RESEARCH LA English DT Article DE glucocorticoid; feedback; repeated stress; tyrosine hydroxylase; glucocorticoid receptor; locus coeruleus ID CORTICOTROPIN-RELEASING FACTOR; MESSENGER-RNA LEVELS; RAT-BRAIN; NORADRENERGIC NEURONS; INSITU HYBRIDIZATION; PARAVENTRICULAR NUCLEUS; SUBTOTAL DESTRUCTION; RESPONSIVE ELEMENT; PROMOTER REGION; FIRING RATE AB Sustained responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis during chronic or repeated stress is associated with continuous activation of ascending noradrenergic neurons from the brainstem to the hypothalamic paraventricular nucleus (PVN). The tact that glucocorticoid receptor (GR) exists in the brainstem noradrenergic neurons including locus coeruleus (LC) suggests that glucocorticoids play a modulatory role in maintaining the activity of these neurons during chronic stress. To determine whether alterations in the sensitivity of noradrenergic neuronal activity to endogenous CORT occur during chronic or repeated stress, tyrosine hydroxylase (TH) and GR mRNA expressions in the LC were examined in acute (2 h) and repeated (2 h daily, 14 days) immobilization stress, using sham-operated rats and adrenalectomized rats with a moderate dose of CORT replacement (ADX+CORT group). In acute stress, TH mRNA in the LC increased in the ADX+CORT rats, but not in sham operated rats. In repeated stress, however, elevated endogenous CORT failed to inhibit TH mRNA responses in sham rats; LC TH mRNA in sham rats responded to the same extent as in ADX+CORT rats. A reduction of GR mRNA in the LC was observed in the acutely stressed and repeatedly stressed sham group, but not in the ADX+CORT groups. The decrease in LC GR mRNA levels in sham rats tended to be greater after repeated than after acute stress. LC GR mRNA levels decreased in response to systemic CORT treatment (200 mg pellet sc, for 14 days) and increased in response to adrenalectomy; neither CORT treatment nor adrenalectomy influenced TH mRNA levels in the LC. These results suggest that glucocorticoid responses to acute immobilization prevent LC TH mRNA levels from rising significantly, while glucocorticoids appear to decrease their capacity to restrain LC TH mRNA during repeated immobilization. Although the results clearly show glucocorticoid-dependent alterations in LC GR mRNA expression, the association between increased TH mRNA and decreased GR mRNA in the LC remains to be elucidated. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. RP Kochi Med Sch, Dept Internal Med 2, Okoh Cho, Nankoku, Kochi 783, Japan. EM fwjf6671@mb.infoweb.ne.jp NR 46 TC 45 Z9 49 U1 0 U2 5 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 EI 1872-6240 J9 BRAIN RES JI Brain Res. PD JUL 12 PY 2002 VL 943 IS 2 BP 216 EP 223 AR PII S0006-8993(02)02647-1 DI 10.1016/S0006-8993(02)02647-1 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 580FE UT WOS:000177223800007 PM 12101044 ER PT J AU Gagnon, E Duclos, S Rondeau, C Chevet, E Cameron, PH Steele-Mortimer, O Paiement, J Bergeron, JJM Desjardins, M AF Gagnon, E Duclos, S Rondeau, C Chevet, E Cameron, PH Steele-Mortimer, O Paiement, J Bergeron, JJM Desjardins, M TI Endoplasmic reticulum-mediated phagocytosis is a mechanism of entry into macrophages SO CELL LA English DT Article ID ISOLATED RAT HEPATOCYTES; LEGIONELLA-PNEUMOPHILA; PHOSPHATIDYLINOSITOL 3-KINASE; MATURING PHAGOSOMES; CONTAINING VACUOLES; IN-VITRO; MATURATION; FUSION; CALRETICULIN; CELLS AB Phagocytosis is a key aspect of our innate ability to fight infectious diseases. In this study, we have found that fusion of the endoplasmic reticulum (ER) with the macrophage plasmalemma, underneath phagocytic cups, is a source of membrane for phagosome formation in macrophages. Successive waves of ER become associated with maturing phagosomes during phagolysosome biogenesis. Thus, the ER appears to possess unexpectedly pluripotent fusion properties. ER-mediated phagocytosis is regulated in part by phosphatidylinositol 3-kinase and used for the internalization of inert particles and intracellular pathogens, regardless of their final trafficking in the host. In neutrophils, where pathogens are rapidly killed, the ER is not used as a major source of membrane for phagocytosis. We propose that intracellular pathogens have evolved to adapt and exploit ER-mediated phagocytosis to avoid destruction in host cells. C1 Univ Montreal, Dept Pathol & Biol Cellulaire, Montreal, PQ H3C 3J7, Canada. McGill Univ, Dept Surg, Montreal, PQ H3A 2T5, Canada. McGill Univ, Dept Anat & Cell Biol, Montreal, PQ H3A 2T5, Canada. NIAID, Rocky Mt Labs, Intracellular Parasites Lab, Hamilton, MT 59840 USA. Caprion Pharmaceut Inc, Montreal, PQ, Canada. RP Desjardins, M (reprint author), Univ Montreal, Dept Pathol & Biol Cellulaire, Montreal, PQ H3C 3J7, Canada. RI Chevet, Eric/E-4992-2016 OI Chevet, Eric/0000-0001-5855-4522 NR 56 TC 459 Z9 471 U1 5 U2 18 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE,, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD JUL 12 PY 2002 VL 110 IS 1 BP 119 EP 131 DI 10.1016/S0092-8674(02)00797-3 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 573PU UT WOS:000176840700012 PM 12151002 ER PT J AU Birnbaumer, L AF Birnbaumer, L TI TRPC4 knockout mice - The coming of age of TRP channels as gates of calcium entry responsible for cellular responses SO CIRCULATION RESEARCH LA English DT Editorial Material DE transient receptor potential; I-CRAC; calcium signaling; phospholipase; endothelial cell function ID DROSOPHILA TRP; MAST-CELLS; CA2+; GENE C1 Natl Inst Environm Hlth Sci, Transmembrane Signaling Grp, Lab Signal Transduct, Res Triangle Pk, NC USA. Natl Inst Environm Hlth Sci, Transmembrane Signaling Grp, Div Intramural Res, Res Triangle Pk, NC USA. RP Birnbaumer, L (reprint author), Bldg 101,Rm A-214,111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 15 TC 12 Z9 12 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7330 J9 CIRC RES JI Circ.Res. PD JUL 12 PY 2002 VL 91 IS 1 BP 1 EP 3 DI 10.1161/01.RES.0000026600.67226.92 PG 3 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 575JX UT WOS:000176944100001 PM 12114313 ER PT J AU Jo, SH Leblais, V Wang, PH Crow, MT Xiao, RP AF Jo, SH Leblais, V Wang, PH Crow, MT Xiao, RP TI Phosphatidylinositol 3-kinase functionally compartmentalizes the concurrent G(s) signaling during beta(2)-adrenergic stimulation SO CIRCULATION RESEARCH LA English DT Article DE beta(2)-adrenoceptor; cAMP signal compartmentation; phosphatidylinositol 3-kinase; phospholamban; cardiac contractility ID CARDIAC MYOCYTES; PROTEIN-KINASE; PHOSPHOINOSITIDE 3-KINASE; HEART-CELLS; PHOSPHOLAMBAN PHOSPHORYLATION; RECEPTOR STIMULATION; CA2+; ACTIVATION; INHIBITOR; CONTRACTILITY AB Compartmentation of intracellular signaling pathways serves as an important mechanism conferring the specificity of G protein-coupled receptor (GPCR) signaling. In the heart, stimulation Of beta(2)-adrenoceptor (beta(2)-AR), a prototypical GPCR, activates a tightly localized protein kinase A (PKA) signaling, which regulates substrates at cell surface membranes, bypassing cytosolic target proteins (ca, phospholamban). Although a concurrent activation of beta(2)-AR-coupled G, proteins has been implicated in the functional compartmentation of PKA signaling, the exact mechanism underlying the restriction of the beta(2)-AR-PKA pathway remains unclear. In the present study, we demonstrate that phosphatidylinositol 3-kinase (PI3K) plays an essential role in confining the beta(2)-AR-PKA signaling. Inhibition of PI3K with LY294002 or wortmannin enables beta(2)-AR-PKA signaling to reach intracellular substrates, as manifested by a robust increase in phosphorylation of phospholamban, and markedly enhances the receptor-mediated positive contractile and relaxant responses in cardiac myocytes. These potentiating effects of PI3K inhibitors are not accompanied by an increase in beta(2)-AR-induced cAMP formation. Blocking G, or GOT signaling with pertussis toxin or betaARK-ct, a peptide inhibitor of GOT, completely prevents the potentiating effects induced by PI3K inhibition, indicating that the pathway responsible for the functional compartmentation Of beta(2)-AR-PKA signaling sequentially involves G(i), Gbetagamma, and PI3K. Thus, PI3K constitutes a key downstream event of beta(2)-AR-G, signaling, which confines and negates the concurrent beta(2)-AR/G(s)-mediated PKA signaling. C1 NIA, Gerontol Res Ctr, Lab Cardiovasc Sci, NIH, Baltimore, MD 21224 USA. Univ Calif Irvine, Dept Med & Biol Chem, Irvine, CA USA. Univ Calif Irvine, Div Endocrinol Diabet & Metab, Irvine, CA USA. RP Xiao, RP (reprint author), NIA, Gerontol Res Ctr, Lab Cardiovasc Sci, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 42 TC 86 Z9 91 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7330 J9 CIRC RES JI Circ.Res. PD JUL 12 PY 2002 VL 91 IS 1 BP 46 EP 53 DI 10.1161/01.RES.0000024115.67561.54 PG 8 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 575JX UT WOS:000176944100009 PM 12114321 ER PT J AU Oganesyan, V Oganesyan, N Terzyan, S Qu, DF Dauter, Z Esmon, NL Esmon, CT AF Oganesyan, V Oganesyan, N Terzyan, S Qu, DF Dauter, Z Esmon, NL Esmon, CT TI The crystal structure of the endothelial protein C receptor and a bound phospholipid SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BLOOD-COAGULATION; BINDING; MECHANISMS; SYSTEM; THROMBOMODULIN; IDENTIFICATION; EXPRESSION; VESICLES; CELLS; MODE AB The endothelial cell protein C receptor (EPCR) shares similar to20% sequence identity with the major histocompatibility complex class 1/CD1 family of molecules, accelerates the thrombin-thrombomodulin-dependent generation of activated protein C, a natural anticoagulant, binds to activated neutrophils, and can undergo translocation from the plasma membrane to the nucleus. Blocking protein C/activated protein C binding to the receptor inhibits not only protein C activation but the ability of the host to respond appropriately to bacterial challenge, exacerbating both the coagulant and inflammatory responses. To understand how EPCR accomplishes these multiple tasks, we solved the crystal structure of EPCR alone and in complex with the phospholipid binding domain of protein C. The structures were strikingly similar to CD1d. A tightly bound phospholipid resides in the groove typically involved in antigen presentation. The protein C binding site is outside this conserved groove and is distal. from the membrane-spanning domain. Extraction of the lipid resulted in loss of protein C binding, which could be restored by lipid reconstitution. CD1d augments the immune response by presenting glycolipid antigens. The EPCR structure is a model for how CD1d binds lipids and further suggests additional potential functions for EPCR in immune regulation, possibly including the anti-phospholipid syndrome. C1 Oklahoma Med Res Fdn, Howard Hughes Med Inst, Oklahoma City, OK 73104 USA. Oklahoma Med Res Fdn, Cardiovasc Biol Res Program, Oklahoma City, OK 73104 USA. Oklahoma Med Res Fdn, Dept Crystallog, Oklahoma City, OK 73104 USA. NCI, SAIC, NIH, Frederick, MD 21702 USA. Univ Oklahoma, Hlth Sci Ctr, Dept Pathol, Oklahoma City, OK 73104 USA. Univ Oklahoma, Hlth Sci Ctr, Dept Biochem & Mol Biol, Oklahoma City, OK 73104 USA. RP Esmon, CT (reprint author), Oklahoma Med Res Fdn, Howard Hughes Med Inst, 825 NE 13th St, Oklahoma City, OK 73104 USA. FU NHLBI NIH HHS [P50 HL54502] NR 27 TC 137 Z9 142 U1 0 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 12 PY 2002 VL 277 IS 28 BP 24851 EP 24854 DI 10.1074/jbc.C200163200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 571YP UT WOS:000176747000003 PM 12034704 ER PT J AU Inoue, Y Hayhurst, GP Inoue, J Mori, M Gonzalez, FJ AF Inoue, Y Hayhurst, GP Inoue, J Mori, M Gonzalez, FJ TI Defective ureagenesis in mice carrying a liver-specific disruption of he-datocvte nuclear factor 4 alpha (HNF4 alpha) - HNF4 alpha regulates ornithine transcarbamylase in vivo SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ENRICHED TRANSCRIPTION FACTORS; HEMOPHILIA-B LEYDEN; FACTOR-IX PROMOTER; GENE-EXPRESSION; BINDING-SITE; CYCLE ENZYMES; VISCERAL ENDODERM; RESPONSE ELEMENT; C/EBP-BETA; CELL AB Hepatocyte nuclear factor 4alpha (HNF4alpha) regulates the expression of many genes preferentially expressed in liver. HNF4alpha-null mice die during embryogenesis precluding the analysis of its function in the adult. To circumvent this problem, liver-specific HNF4alpha-null mice were produced. Mice lacking hepatic HNF4alpha expression exhibited increased serum ammonia and reduced serum urea. This disruption in ureagenesis may be explained by a marked decrease in expression and activity of hepatic ornithine transcarbamylase (OTC). To determine the molecular mechanisms involved in transcriptional regulation of the mouse OTC gene, the OTC promoter region was analyzed. Sequence analysis revealed the presence of two putative HNF4alpha-binding sites in the mouse OTC promoter region. By using transient transfection analysis, it was established that high levels of promoter activity were dependent on both HNF4alpha-binding sites and the expression of HNF4alpha. Furthermore, the proximal HNF4alpha-binding site was found to be more important than the distal. one for transactivating OTC promoter. These data demonstrate that HNF4alpha is critical for urea homeostasis by direct regulation of the OTC gene in vivo. C1 NCI, Div Basic Sci, Lab Metab, NIH, Bethesda, MD 20892 USA. Kumamoto Univ, Sch Med, Dept Mol Genet, Kumamoto 8600811, Japan. RP Gonzalez, FJ (reprint author), NCI, Div Basic Sci, Lab Metab, NIH, 9000 Rockville Pike,Rm 3E-24,Bldg 37, Bethesda, MD 20892 USA. NR 57 TC 61 Z9 61 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 12 PY 2002 VL 277 IS 28 BP 25257 EP 25265 DI 10.1074/jbc.M203126200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 571YP UT WOS:000176747000054 PM 11994307 ER PT J AU Chang, TS Jeong, W Choi, SY Yu, SQ Kang, SW Rhee, SG AF Chang, TS Jeong, W Choi, SY Yu, SQ Kang, SW Rhee, SG TI Regulation of peroxiredoxin I activity by Cdc2-mediated phosphorylation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CYCLIN-DEPENDENT KINASES; TYROSINE-PHOSPHATASE 1B; MAMMALIAN PEROXIREDOXIN; HYDROGEN-PEROXIDE; CELL-CYCLE; CRYSTAL-STRUCTURE; OKADAIC ACID; REVERSIBLE INACTIVATION; THIOREDOXIN PEROXIDASE; PROTEIN PHOSPHATASES AB Hydrogen peroxide is implicated as an intracellular messenger in various cellular responses such as proliferation and differentiation. Peroxiredoxin (Prx) I is a member of the peroxiredoxin family of peroxidases and contains a consensus site (Thr(90)-Pro-Lys-Lys) for phosphorylation by cyclin-dependent kinases (CDKs). This protein has now been shown to be phosphorylated specifically on Thr(90) by several CDKs, including Cdc2, in vitro. Phosphorylation of Prx I on Thr(90) reduced the peroxidase activity of this protein by 80%. The phosphorylation of Prx I in HeLa cells was monitored with the use of antibodies specific for Prx I phosphorylated on Thr(90). Immunoblot analysis with these antibodies of HeLa cells arrested at various stages of the cell cycle revealed that Prx I phosphorylation occurs in parallel with the activation of Cdc2; Prx I phosphorylation was thus marked during mitosis but virtually undetectable during interphase. Furthermore, when Cdc2 expression was reduced by RNA interference with cognate small interfering RNAs, Prx I phosphorylation was not observed in the cells synchronized in mitotic phase. The cytosolic location of Prx I likely prevents its interaction with activated CDKs until after the breakdown of the nuclear envelope during mitosis, when Cdc2 is the CDK that is most active. Phosphorylation of Prx I on Thr90 both in vitro and in vivo was blocked by roscovitine, an inhibitor of CDKs. These results suggest that Cdc2-mediated phosphorylation and inactivation of Prx I and the resulting intracellular accumulation of H2O2 might be important for progression of the cell cycle. C1 NHLBI, Lab Cell Signaling, NIH, Bethesda, MD 20892 USA. Ewha Womans Univ, Ctr Cell Signaling Res, Seoul 120750, South Korea. RP Rhee, SG (reprint author), Bldg 50,Rm 3523,S Dr,MSC 8015, Bethesda, MD 20892 USA. NR 55 TC 168 Z9 187 U1 1 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 12 PY 2002 VL 277 IS 28 BP 25370 EP 25376 DI 10.1074/jbc.M110432200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 571YP UT WOS:000176747000069 PM 11986303 ER PT J AU Greene, BT Thorburn, J Willingham, MC Thorburn, A Planalp, RP Brechbiel, MW Jennings-Gee, J Wilkinson, J Torti, FM Torti, SV AF Greene, BT Thorburn, J Willingham, MC Thorburn, A Planalp, RP Brechbiel, MW Jennings-Gee, J Wilkinson, J Torti, FM Torti, SV TI Activation of caspase pathways during iron chelator-mediated apoptosis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DRUG-INDUCED APOPTOSIS; ISONICOTINOYL HYDRAZONE; NEUROBLASTOMA-CELLS; SIGNALING PATHWAY; NECROSIS-FACTOR; CYTOCHROME-C; PROTEIN P35; DEPRIVATION; CIS,CIS-1,3,5-TRIAMINOCYCLOHEXANE; INHIBITION AB Iron chelators have traditionally been used in the treatment of iron overload. Recently, chelators have also been explored for their ability to limit oxidant damage in cardiovascular, neurologic, and inflammatory disease as well as to serve as anti-cancer agents. To determine the mechanism of cell death induced by iron chelators, we assessed the time course and pathways of caspase activation during apoptosis induced by iron chelators. We report that the chelator tachpyridine sequentially activates caspases 9, 3, and 8. These caspases were also activated by the structurally unrelated chelators dipyridyl and desferrioxamine. The critical role of caspase activation in cell death was supported by micro-injection experiments demonstrating that p35, a broad spectrum caspase inhibitor, protected HeLa cells from chelator-induced cell death. Apoptosis mediated by tachpyridine was not prevented by blocking the CD95 death receptor pathway with a Fas-associated death domain protein (FADD) dominant-negative mutant. In contrast, chelator-mediated cell death was blocked in cells microinjected with Bcl-XL and completely inhibited in cells microinjected with a dominant-negative caspase 9 expression vector. Caspase activation was not observed in cells treated with N-methyl tachpyridine, an N-alkylated derivative of tachpyridine which lacks an ability to react with iron. These results suggest that activation of a mitochondrial caspase pathway is an important mechanism by which iron chelators induce cell death. C1 Wake Forest Univ, Sch Med, Dept Biochem, Winston Salem, NC 27157 USA. Wake Forest Univ, Sch Med, Dept Pathol, Winston Salem, NC 27157 USA. Wake Forest Univ, Sch Med, Dept Biochem, Winston Salem, NC 27157 USA. Wake Forest Univ, Sch Med, Ctr Comprehens Canc, Winston Salem, NC 27157 USA. Univ New Hampshire, Dept Chem, Durham, NH 03824 USA. NIH, Radiat Oncol Branch, Bethesda, MD 20892 USA. RP Torti, SV (reprint author), Wake Forest Univ, Sch Med, Dept Biochem, Hanes Bldg,Rm 4057,Med Ctr Blvd, Winston Salem, NC 27157 USA. FU NIDDK NIH HHS [DK 57781] NR 45 TC 61 Z9 65 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 12 PY 2002 VL 277 IS 28 BP 25568 EP 25575 DI 10.1074/jbc.M110345200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 571YP UT WOS:000176747000094 PM 11980894 ER PT J AU Du, QZ Xia, M Ito, Y AF Du, QZ Xia, M Ito, Y TI Purification of icariin from the extract of Epimedium segittatum using high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE Epimedium segittatum; counter-current chromatography; icariin AB Icariin was purified from the extract of Epimedium segittatum by high-speed counter-current chromatography with a two-phase solvent system composed of n-hexane-n-butanol-methanol-water (1:4:2:6, v/v). We used two multilayer coil planet centrifuges with different capacities. A 300 mg amount of the extract was separated using a semipreparative instrument equipped with a 230-ml capacity column to yield 103 mg of icariin at 86.2% purity. An 8 g amount of the extract was separated with a large preparative instrument equipped with a 2460-ml capacity column to produce 2.45 g of icariin at 85.7% purity. From these fractions over 98% pure icariin was obtained by recrystallization with water. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NHLBI, Lab Biophys Chem, NIH, Bethesda, MD 20892 USA. Hangzhou Univ Commerce, Inst Food & Biol Engn, Hangzhou 310035, Peoples R China. RP Ito, Y (reprint author), NHLBI, Lab Biophys Chem, NIH, Bldg 50,Rm 3334,50 S Dr MSC, Bethesda, MD 20892 USA. NR 7 TC 28 Z9 40 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD JUL 12 PY 2002 VL 962 IS 1-2 BP 239 EP 241 AR PII S0021-9673(02)00538-1 DI 10.1016/S0021-9673(02)00538-1 PG 3 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 577LB UT WOS:000177061400022 PM 12198968 ER PT J AU Ma, XF Zhang, TY Wei, Y Tu, PF Chen, YJ Ito, Y AF Ma, XF Zhang, TY Wei, Y Tu, PF Chen, YJ Ito, Y TI Preparative isolation and purification of calycosin from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE Astragalus mongholicus; preparative chromatography; counter-current chromatography; calycosin ID SEPARATION; FLAVONOIDS AB Calycosin was purified from an ethyl acetate extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography. The separation was performed in two steps with a two-phase solvent system composed of n-hexane-chloroforrn-methanol-water (1:3:3:2, v/v). From 200 mg of the crude extract, 14.8 mg of calycosin was obtained at over 99% purity as determined by HPLC analysis, and its chemical structure was confirmed by MS,H-1 and 13 degreesC nuclear magnetic resonance. Published by Elsevier Science B.V. C1 NHLBI, Lab Biophys Chem, NIH, Bethesda, MD 20892 USA. Peking Univ, Modern Res Ctr Traditional Chinese Med, Beijing 100083, Peoples R China. Beijing Inst New Technol Applicat, Beijing 100035, Peoples R China. Shenyang Pharmaceut Univ, Shenyang 110016, Peoples R China. RP Ito, Y (reprint author), NHLBI, Lab Biophys Chem, NIH, Bldg 50,Rm 3334,50 S Dr MSC 8014, Bethesda, MD 20892 USA. EM itoy@nhlbi.nih.gov NR 18 TC 24 Z9 27 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD JUL 12 PY 2002 VL 962 IS 1-2 BP 243 EP 247 AR PII S0021-9673(02)00535-6 DI 10.1016/S0021-9673(02)00535-6 PG 5 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 577LB UT WOS:000177061400023 PM 12198969 ER PT J AU Dimasi, N Sawicki, MW Reineck, LA Li, YL Natarajan, K Margulies, DH Mariuzza, RA AF Dimasi, N Sawicki, MW Reineck, LA Li, YL Natarajan, K Margulies, DH Mariuzza, RA TI Crystal structure of the Ly49I natural killer cell receptor reveals variability in dimerization mode within the Ly49 family SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE natural killer cells; NK cell receptors; MHC class I; X-ray crystallography ID C-TYPE LECTIN; CLASS-I LIGAND; BINDING PROTEIN; GENE FAMILY; RECOGNITION; DOMAIN; IDENTIFICATION; MOLECULES; PROGRAM; SYSTEM AB Natural killer (NK) cells play a crucial role in the detection and destruction of virally infected and tumor cells during innate immune responses. The cytolytic activity of NK cells is regulated through a balance of inhibitory and stimulatory signals delivered by NK receptors that recognize classical major histocompatabilty complex class I (MHC-I) molecules, or MHC-I homologs such as MICA, on target cells. The Ly49 family of NK receptors (Ly49A through W), which includes both inhibitory and activating receptors, are homodimeric type II transmembrane glycoproteins, with each subunit composed of a C-type lectin-like domain tethered to the membrane by a stalk region. We have determined the crystal structure, at 3.0 Angstrom resolution, of the murine inhibitory NK receptor Ly491. The Ly491 monomer adopts a fold similar to that of other C-type lectin-like NK receptors, including Ly49A, NKG2D and CD69. However, the Ly491 monomers associate in a manner distinct from that of these other NK receptors, forming a more open dimer. As a result, the putative MHC-binding surfaces of the Ly491 dimer are spatially more distant than the corresponding surfaces of Ly49A or NKG2D. These structural differences probably reflect the fundamentally different ways in which Ly49 and NKG2D receptors recognize their respective ligands: whereas the single MICA binding site of NKG2D is formed by the precise juxtaposition of two monomers, each Ly49 monomer contains an independent binding site for MHC-I. Hence, the structural constraints on dimerization geometry may be relatively relaxed within the Ly49 family. Such variability may enable certain Ly49 receptors, like Ly491, to bind MHC-I molecules bivalently, thereby stabilizing receptor-ligand interactions and enhancing signal transmission to the NK cell. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Univ Maryland, Maryland Biotechnol Inst, Ctr Adv Res Biotechnol, WM Keck Lab Struct Biol, Rockville, MD 20850 USA. NIAID, Mol Biol Sect, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Mariuzza, RA (reprint author), Univ Maryland, Maryland Biotechnol Inst, Ctr Adv Res Biotechnol, WM Keck Lab Struct Biol, 9600 Gudelsky Dr, Rockville, MD 20850 USA. RI Margulies, David/H-7089-2013; OI Margulies, David/0000-0001-8530-7375 FU NIAID NIH HHS [AI36900, AI47900] NR 42 TC 23 Z9 23 U1 0 U2 2 PU ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD JUL 12 PY 2002 VL 320 IS 3 BP 573 EP 585 DI 10.1016/S0022-2836(02)00498-9 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 578CB UT WOS:000177098900010 PM 12096910 ER PT J AU Bosetti, F Seemann, R Rapoport, SI AF Bosetti, F Seemann, R Rapoport, SI TI Chronic lithium chloride administration to rats decreases brain protein level of epsilon (epsilon) subunit of eukaryotic initiation factor-2B SO NEUROSCIENCE LETTERS LA English DT Article DE lithium; brain; rat; eukaryotic initiation factor-28; translation; bipolar disorder; chronic; transcription; protein synthesis ID FACTOR 2B; ADENYLATE-CYCLASE; ALPHA-SUBUNIT; PHOSPHORYLATION; EXPRESSION; RECEPTORS; SYSTEM; CELLS AB The eukaryotic initiation factor-2B (eIF-2B) can regulate translation and protein synthesis. We used Western blot analysis to quantify the protein level of the catalytic epsilon (epsilon) subunit of eIF-2B in brains of rats fed lithium chloride (LiCl) for 6 weeks so as to produce a brain lithium concentration that is therapeutically effective in bipolar disorder. The ratio of eIF-2B (epsilon) to actin protein was significantly reduced (P < 0.01) in LiCl-fed rats, 0.86 +/- 0.06 (SE) compared to 1.2 +/- 0.07 in control rats. These results suggest that a therapeutic level of lithium may downregulate the synthesis of proteins whose translation depends on eIF-2B. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. RP Bosetti, F (reprint author), NIA, Brain Physiol & Metab Sect, NIH, 9000 Rockville Pike,Bldg 10,Room 6N202, Bethesda, MD 20892 USA. NR 26 TC 7 Z9 7 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD JUL 12 PY 2002 VL 327 IS 1 BP 71 EP 73 AR PII S0304-3940(02)00354-3 DI 10.1016/S0304-3940(02)00354-3 PG 3 WC Neurosciences SC Neurosciences & Neurology GA 581BW UT WOS:000177272100018 PM 12098503 ER PT J AU Hwang, JJ Allen, PD Tseng, GC Lam, CW Fananapazir, L Dzau, VJ Liew, CC AF Hwang, JJ Allen, PD Tseng, GC Lam, CW Fananapazir, L Dzau, VJ Liew, CC TI Microarray gene expression profiles in dilated and hypertrophic cardiomyopathic end-stage heart failure SO PHYSIOLOGICAL GENOMICS LA English DT Article; Proceedings Paper CT 5th Annual Meeting of the Heart-Failure-Society-of-America CY SEP 09-12, 2001 CL WASHINGTON, D.C. SP Heart Failure Soc Amer DE cDNA microarray; normalization; real-time reverse transcription-polymerase chain reaction ID MESSENGER-RNA EXPRESSION; CARDIAC-HYPERTROPHY; CDNA MICROARRAY; CARDIOVASCULAR-DISEASE; ANNEXIN-II; PROTEINS; PATTERNS; CANCER; CELL; IDENTIFICATION AB Despite similar clinical endpoints, heart failure resulting from dilated cardiomyopathy (DCM) or hypertrophic cardiomyopathy (HCM) appears to develop through different remodeling and molecular pathways. Current understanding of heart failure has been facilitated by microarray technology. We constructed an in-house spotted cDNA microarray using 10,272 unique clones from various cardiovascular cDNA libraries sequenced and annotated in our laboratory. RNA samples were obtained from left ventricular tissues of precardiac transplantation DCM and HCM patients and were hybridized against normal adult heart reference RNA. After filtering, differentially expressed genes were determined using novel analyzing software. We demonstrated that normalization for cDNA microarray data is slide-dependent and nonlinear. The feasibility of this model was validated by quantitative real-time reverse transcription-PCR, and the accuracy rate depended on the fold change and statistical significance level. Our results showed that 192 genes were highly expressed in both DCM and HCM (e. g., atrial natriuretic peptide, CD59, decorin, elongation factor 2, and heat shock protein 90), and 51 genes were downregulated in both conditions (e. g., elastin, sarcoplasmic/endoplasmic reticulum Ca2+-ATPase). We also identified several genes differentially expressed between DCM and HCM (e. g., alphaB-crystallin, antagonizer of myc transcriptional activity, beta-dystrobrevin, calsequestrin, lipocortin, and lumican). Microarray technology provides us with a genomic approach to explore the genetic markers and molecular mechanisms leading to heart failure. C1 Harvard Univ, Sch Med, Brigham & Womens Hosp, Cardiovasc Genome Unit,Dept Med, Boston, MA 02115 USA. Harvard Univ, Sch Med, Brigham & Womens Hosp, Dept Anesthesiol, Boston, MA 02115 USA. Natl Taiwan Univ Hosp, Dept Internal Med, Div Cardiovasc Med, Taipei 100, Taiwan. Harvard Univ, Sch Publ Hlth, Dept Biostat, Boston, MA 02115 USA. NHLBI, Cardiol Branch, Bethesda, MD 20892 USA. Univ Toronto, Dept Lab Med & Pathol, Toronto, ON, Canada. Chinese Univ Hong Kong, Prince Wales Hosp, Dept Chem Pathol, Hong Kong, Hong Kong, Peoples R China. RP Liew, CC (reprint author), Harvard Univ, Sch Med, Brigham & Womens Hosp, Cardiovasc Genome Unit,Dept Med, 75 Francis St,Thorn 1326, Boston, MA 02115 USA. EM cliew@rics.bwh.harvard.edu OI Juey-Jen, HWANG/0000-0001-6437-0455 FU NHLBI NIH HHS [5R37-HL-3561016, 5P5O-HL-5931603, 5RO1-HL-5851603, 5RO1-HL-6166102] NR 54 TC 157 Z9 160 U1 0 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1094-8341 J9 PHYSIOL GENOMICS JI Physiol. Genomics PD JUL 12 PY 2002 VL 10 IS 1 BP 31 EP 44 DI 10.1152/physiolgenomics.00122.2001. PG 14 WC Cell Biology; Genetics & Heredity; Physiology SC Cell Biology; Genetics & Heredity; Physiology GA 571ZL UT WOS:000176749000005 PM 12118103 ER PT J AU Scaffidi, P Misteli, T Bianchi, ME AF Scaffidi, P Misteli, T Bianchi, ME TI Release of chromatin protein HMGB1 by necrotic cells triggers inflammation SO NATURE LA English DT Article ID LINKER HISTONES B4; HIGH-MOBILITY; CHROMOSOMAL-PROTEINS; DNA; BINDING; H1; APOPTOSIS AB High mobility group 1 (HMGB1) protein is both a nuclear factor and a secreted protein. In the cell nucleus it acts as an architectural chromatin-binding factor that bends DNA and promotes protein assembly on specific DNA targets(1,2). Outside the cell, it binds with high affinity to RAGE (the receptor for advanced glycation end products) 3 and is a potent mediator of inflammation(4-6). HMGB1 is secreted by activated monocytes and macrophages(4), and is passively released by necrotic or damaged cells(7-9). Here we report that Hmgb1(-/-) necrotic cells have a greatly reduced ability to promote inflammation, which proves that the release of HMGB1 can signal the demise of a cell to its neighbours. Apoptotic cells do not release HMGB1 even after undergoing secondary necrosis and partial autolysis, and thus fail to promote inflammation even if not cleared promptly by phagocytic cells. In apoptotic cells, HMGB1 is bound firmly to chromatin because of generalized underacetylation of histone and is released in the extracellular medium (promoting inflammation) if chromatin deacetylation is prevented. Thus, cells undergoing apoptosis are programmed to withhold the signal that is broadcast by cells that have been damaged or killed by trauma. C1 Univ Vita Salute San Raffaele, I-20132 Milan, Italy. San Raffaele Sci Inst, DIBIT, I-20132 Milan, Italy. NCI, NIH, Bethesda, MD 20892 USA. RP Bianchi, ME (reprint author), Univ Vita Salute San Raffaele, I-20132 Milan, Italy. OI Bianchi, Marco Emilio/0000-0002-5329-6445 NR 24 TC 2114 Z9 2240 U1 13 U2 112 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 11 PY 2002 VL 418 IS 6894 BP 191 EP 195 DI 10.1038/nature00858 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 571GV UT WOS:000176710400042 PM 12110890 ER PT J AU Chi, TH Wan, M Zhao, KJ Taniuchi, I Chen, L Littman, DR Crabtree, GR AF Chi, TH Wan, M Zhao, KJ Taniuchi, I Chen, L Littman, DR Crabtree, GR TI Reciprocal regulation of CD4/CD8 expression by SWI/SNF-like BAF complexes SO NATURE LA English DT Article ID CD4 GENE-EXPRESSION; T-CELL DEVELOPMENT; STOCHASTIC MECHANISM; CHROMATIN; LINEAGE; PROMOTER; SUBSETS; BRAHMA; YEAST AB Thymic development produces two sub-lineages of T cells expressing either CD4 or CD8 co-receptors that assist antibody production and mediate cell killing, respectively. The mechanisms for mutually exclusive co-receptor expression remain poorly defined(1,2). We find that mutations in the high mobility group (HMG) domain of BAF57-a DNA-binding subunit of the mammalian SWI/SNF-like chromatin-remodelling BAF complexesor in the BAF complex ATPase subunit Brg, impair both CD4 silencing and CD8 activation. Brg is haploinsufficient for CD8 activation, but not for CD4 silencing, whereas BAF57 mutations preferentially impair CD4 silencing, pointing to target- and subunit-specific mechanisms of chromatin remodelling. BAF complexes directly bind the CD4 silencer, but the BAF57 HMG domain is dispensable for tethering BAF complexes to the CD4 silencer or other chromatin loci in vivo, or for remodelling reconstituted templates in vitro(3,4), suggesting that chromatin remodelling in vivo requires HMG-dependent DNA bending. These results indicate that BAF complexes contribute to lineage bifurcation by reciprocally regulating lineage-specific genes, reminiscent of the role of the yeast SWI/SNF complex in mediating mating-type switching(5,6) C1 Stanford Univ, Sch Med, Howard Hughes Med Inst, Dept Pathol & Dev Biol, Stanford, CA 94305 USA. NHLBI, Mol Immunol Lab, NIH, Bethesda, MD 20892 USA. NYU, Sch Med, Skirball Inst Biomol Med, Mol Pathogenesis Program, New York, NY 10016 USA. NYU, Sch Med, Howard Hughes Med Inst, New York, NY 10016 USA. RP Crabtree, GR (reprint author), Stanford Univ, Sch Med, Howard Hughes Med Inst, Dept Pathol & Dev Biol, Stanford, CA 94305 USA. RI Taniuchi, Ichiro/N-6399-2015 OI Taniuchi, Ichiro/0000-0002-9853-9068 NR 23 TC 172 Z9 178 U1 0 U2 7 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 11 PY 2002 VL 418 IS 6894 BP 195 EP 199 DI 10.1038/nature00876 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 571GV UT WOS:000176710400043 PM 12110891 ER PT J AU Gahl, WA Thoene, JG Schneider, JA AF Gahl, WA Thoene, JG Schneider, JA TI Cystinosis SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Review ID NEPHROPATHIC CYSTINOSIS; CYSTEAMINE THERAPY; COUNTER-TRANSPORT; CORNEAL CRYSTALS; BINDING-PROTEIN; CTNS MUTATIONS; RENAL-FUNCTION; GENE CTNS; CHILDREN; INSUFFICIENCY C1 NICHHD, Heritable Disorders Branch, Bethesda, MD 20892 USA. Tulane Hlth Sci Ctr, Hayward Genet Ctr, New Orleans, LA USA. Univ Calif San Diego, Dept Pediat, La Jolla, CA 92093 USA. RP Gahl, WA (reprint author), NICHHD, Heritable Disorders Branch, 10 Ctr Dr,MSC 1830,Bldg 10,Rm 9S-241, Bethesda, MD 20892 USA. FU NCRR NIH HHS [M01 RR00827] NR 74 TC 272 Z9 278 U1 3 U2 16 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUL 11 PY 2002 VL 347 IS 2 BP 111 EP 121 DI 10.1056/NEJMra020552 PG 11 WC Medicine, General & Internal SC General & Internal Medicine GA 572HC UT WOS:000176767300007 PM 12110740 ER PT J AU Horng, S Miller, FG AF Horng, S Miller, FG TI Is placebo surgery unethical? SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material ID CLINICAL RESEARCH; TRIALS; CONSENT C1 NIH, Bethesda, MD 20892 USA. RP Horng, S (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 21 TC 96 Z9 97 U1 0 U2 1 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUL 11 PY 2002 VL 347 IS 2 BP 137 EP 139 DI 10.1056/NEJMsb021025 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 572HC UT WOS:000176767300011 PM 12110744 ER PT J AU Slater, DJ Hilgenfeld, E Rappaport, EF Shah, N Meek, RG Williams, WR Lovett, BD Osheroff, N Autar, RS Ried, T Felix, CA AF Slater, DJ Hilgenfeld, E Rappaport, EF Shah, N Meek, RG Williams, WR Lovett, BD Osheroff, N Autar, RS Ried, T Felix, CA TI MLL-SEPTIN6 fusion recurs in novel translocation of chromosomes 3, X, and 11 in infant acute myelomonocytic leukaemia and in t(X;11) in infant acute myeloid leukaemia, and MLL genomic breakpoint in complex MLL-SEPTIN6 rearrangement is a DNA topoisomerase II cleavage site SO ONCOGENE LA English DT Article DE MLL; SEPTIN6; complex translocation; infant leukaemia; spectral karyotype; DNA topoisomerase II ID ACUTE LYMPHOBLASTIC-LEUKEMIA; CHILDRENS CANCER GROUP; PARTNER GENE; MYELODYSPLASTIC SYNDROME; T(4-11) TRANSLOCATION; DROSOPHILA-TRITHORAX; AF-4 GENE; FUSES MLL; IDENTIFICATION; SEPTINS AB We examined the MLL translocation in two cases of infant AML with X chromosome disruption. The G-banded karyotype in the first case suggested t(X;3)(q22;p21)ins(X;11)(q22;q13q25). Southern blot analysis showed one MLL rearrangement. Panhandle PCR approaches were used to identify the MLL fusion transcript and MLL genomic breakpoint junction. SEPTIN6 from chromosome band Xq24 was the partner gene of MLL. MLL exon 7 was joined in-frame to SEPTIN6 exon 2 in the fusion transcript. The MLL genomic breakpoint was in intron 7; the SEPTIN6 genomic breakpoint was in intron 1. Spectral karyotyping revealed a complex rearrangement disrupting band 11q23. FISH with a probe for MLL confirmed MLL involvement and showed that the MLL-SEPTIN6 junction was on the der(X). The MLL genomic breakpoint was a functional DNA topoisomerase 11 cleavage site in an in vitro assay. In the second case, the karyotype revealed t(X;11)(q22;q23). Southern blot analysis showed two MLL rearrangements. cDNA panhandle PCR detected a transcript fusing MLL exon 8 in-frame to SEPTIN6 exon 2. MLL and SEPTIN6 are vulnerable to damage to form recurrent translocations in infant AML. Identification of SEPTIN6 and the SEPTIN family members hCDCrel and MSF as partner genes of MLL suggests a common pathway to leukaemogenesis. C1 Childrens Hosp Philadelphia, Div Oncol, Abramson Res Ctr, Philadelphia, PA 19104 USA. Childrens Hosp Philadelphia, Joseph Stokes Jr Res Inst, Philadelphia, PA 19104 USA. Univ Penn, Dept Pediat, Sch Med, Philadelphia, PA 19104 USA. NCI, Div Genet, Bethesda, MD 20892 USA. Geisinger Med Ctr, Div Hematol Oncol, Dept Pediat, Danville, PA 17822 USA. Alfred I Dupont Inst, Dept Hematol Oncol, Dept Pediat, Wilmington, DE 19899 USA. Vanderbilt Univ, Sch Med, Dept Biochem, Nashville, TN 37232 USA. RP Felix, CA (reprint author), Childrens Hosp Philadelphia, Div Oncol, Abramson Res Ctr, Rm 902B,3516 Civ Ctr Blvd, Philadelphia, PA 19104 USA. FU NCI NIH HHS [CA85469, CA77683, CA80175]; NIGMS NIH HHS [GM33944] NR 72 TC 43 Z9 45 U1 1 U2 5 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUL 11 PY 2002 VL 21 IS 30 BP 4706 EP 4714 DI 10.1038/sj.onc.1205572 PG 9 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 569WH UT WOS:000176625100013 PM 12096348 ER PT J AU Hall, D Minton, AP AF Hall, D Minton, AP TI Effects of inert volume-excluding macromolecules on protein fiber formation. I. Equilibrium models SO BIOPHYSICAL CHEMISTRY LA English DT Article DE protein assembly; macromolecular crowding; excluded volume; thermodynamic nonideality ID LINEAR SELF-ASSOCIATION; POLYMERIZATION; TERMS; FTSZ AB The equilibrium Oosawa-Asakura model for nucleated assembly of rod-like protein fibers is recast in terms of dimensionless (scaled) quantities. The model is then generalized to treat arbitrarily large deviations from thermodynamic ideality arising from high fractional volume occupancy by an inert protein or polymer. Each state of association of the self-associating protein is modeled as an equivalent rigid convex particle (sphere or spherocylinder) and the crowding species is modeled either as an equivalent sphere or cylindrical rod. The resulting conservation of mass relation is readily solved to yield the fractional abundance of monomer, from which the entire equilibrium distribution of oligomeric species can be calculated, either directly or through the use of an additional scaling relationship. Results indicating the potential effect of volume occupancy on the equilibrium solubility of the self-associating protein and upon the equilibrium distribution of polymer size are presented. It is found that the fractional (logarithmic) change in both solubility and in the breadth of the polymer size distribution scale almost linearly with the fractional (logarithmic) change in the thermodynamic activity of monomer. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Sect Phys Biochem, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Minton, AP (reprint author), NIDDKD, Sect Phys Biochem, Lab Biochem & Genet, NIH, Bldg 8,Room 226, Bethesda, MD 20892 USA. RI Hall, Damien/D-9927-2012 OI Hall, Damien/0000-0003-1538-7618 NR 24 TC 21 Z9 21 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0301-4622 J9 BIOPHYS CHEM JI Biophys. Chem. PD JUL 10 PY 2002 VL 98 IS 1-2 BP 93 EP 104 AR PII S0301-4622(02)00087-X DI 10.1016/S0301-4622(02)00087-X PG 12 WC Biochemistry & Molecular Biology; Biophysics; Chemistry, Physical SC Biochemistry & Molecular Biology; Biophysics; Chemistry GA 584CQ UT WOS:000177450000009 PM 12128192 ER PT J AU Henry, ER Bettati, S Hofrichter, J Eaton, WA AF Henry, ER Bettati, S Hofrichter, J Eaton, WA TI A tertiary two-state allosteric model for hemoglobin SO BIOPHYSICAL CHEMISTRY LA English DT Article DE allostery; hemoglobin; quaternary structure; oxygen binding; cooperative binding; conformational kinetics; conformational substates; stretched exponential kinetics ID CONFORMATIONAL-CHANGES; OXYGEN-AFFINITY; LIGAND-BINDING; ROOM-TEMPERATURE; SINGLE-CRYSTALS; COOPERATIVITY; KINETICS; MYOGLOBIN; EFFECTORS; MECHANISM AB The two-state allosteric model of Monod, Wyman, and Changeux (MWC) provides an excellent description of homotropic effects in a vast array of equilibrium and kinetic measurements on cooperative ligand binding by hemoglobin. However, in contrast to experimental observations, the model does not allow for alteration of the ligand affinity of the T quaternary structure by allosteric effectors. This failure to explain heterotropic effects has been appreciated for over 30 years, and it has been generally assumed to result from tertiary conformational changes in the absence of a quaternary change. Here we explore a model that preserves the essential MWC idea that binding without a quaternary conformational change is non-cooperative, but where tertiary conformations of individual subunits play the primary role instead of the quaternary conformations. In this model, which we call the 'tertiary two-state (TTS) model', the two affinity states correspond to two tertiary conformations of individual subunits rather than the two quaternary conformations of the MWC two-state allosteric model. Ligation and the R quaternary structure bias the subunit population toward the high affinity tertiary conformation, while deligation and the T quaternary structure favor the low affinity tertiary conformation. We show that the model is successful in quantitatively explaining a demanding set of kinetic data from nanosecond carbon monoxide photodissociation experiments at times longer than similar to1 mus. Better agreement between the model and the submicrosecond kinetic data may result from detailed considerations of the distribution and dynamics of conformational substates of the two tertiary conformations. The model is consistent with the results of solution, gel, and single crystal oxygen binding studies, but underestimates the population of doubly-liganded molecules determined in low-temperature electrophoresis experiments. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Eaton, WA (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5, Bethesda, MD 20892 USA. RI Henry, Eric/J-3414-2013 OI Henry, Eric/0000-0002-5648-8696 NR 48 TC 80 Z9 83 U1 2 U2 15 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0301-4622 J9 BIOPHYS CHEM JI Biophys. Chem. PD JUL 10 PY 2002 VL 98 IS 1-2 BP 149 EP 164 AR PII S0301-4622(02)00091-1 DI 10.1016/S0301-4622(02)00091-1 PG 16 WC Biochemistry & Molecular Biology; Biophysics; Chemistry, Physical SC Biochemistry & Molecular Biology; Biophysics; Chemistry GA 584CQ UT WOS:000177450000013 PM 12128196 ER PT J AU Swope, DL Harrell, JC Mahato, D Korach, KS AF Swope, DL Harrell, JC Mahato, D Korach, KS TI Genomic structure and identification of a truncated variant message of the mouse estrogen receptor alpha gene SO GENE LA English DT Article DE estradiol; splice variant; steroid hormone receptor; transcription ID MESSENGER-RNA VARIANTS; HUMAN BREAST-CANCER; TRANSCRIPTIONAL ACTIVATION; REPRODUCTIVE PHENOTYPES; LIGAND-BINDING; EXPRESSION; BETA; TISSUES; ORGANIZATION; MICE AB Estrogen receptor alpha (ERalpha) is a ligand-dependent transcription factor that directs the transcription of a wide number of estrogen-regulated genes. ERalpha mediates the effects of 17-beta-estradiol in both males and females, and was the first estrogen receptor identified. Despite the cloning of the mouse ERalpha cDNA over 15 years ago, the precise genomic organization of the mouse ERalpha gene has not yet been elucidated. In order to determine the structure of this gene, overlapping BAC and P1 clones containing partial genomic sequences of the mouse ERalpha cDNA were obtained from a mouse ES cell genomic library. Using standard restriction fragment analysis followed by Southern blotting, the mouse ERalpha gene was determined to be greater than 220 kb in length. The introns vary widely in size, from 1.8 to 60 kb in length. Sequencing of intron-exon boundaries shows that these boundaries are highly conserved between the human and mouse ERalpha genes. Additionally, we have identified a splice variant message of mouse ERalpha arising from a failure to properly splice at the 3' end of exon 4; the resulting message is predicted to produce protein lacking the ligand-binding domain. Variant message was detected by RT-PCR in several tissues, including uterus, ovary, mammary gland, placenta and testis. Published by Elsevier Science B.V. C1 Natl Inst Environm Hlth Sci, Lab Reproduct & Dev Toxicol, Receptor Biol Sect, Res Triangle Pk, NC 27709 USA. RP Swope, DL (reprint author), Natl Inst Environm Hlth Sci, Lab Reproduct & Dev Toxicol, Receptor Biol Sect, 111 TW Alexander Dr,Rm E460, Res Triangle Pk, NC 27709 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 30 TC 10 Z9 10 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD JUL 10 PY 2002 VL 294 IS 1-2 BP 239 EP 247 AR PII S0378-1119(02)00796-5 DI 10.1016/S0378-1119(02)00796-5 PG 9 WC Genetics & Heredity SC Genetics & Heredity GA 608PK UT WOS:000178855300025 PM 12234686 ER PT J AU Blankson, JN Gallant, JE Quinn, TC Bartlett, JG Siliciano, RF AF Blankson, JN Gallant, JE Quinn, TC Bartlett, JG Siliciano, RF TI Loss of HIV-1-specific immunity during treatment interruption in 2 chronically infected patients SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter ID HIV-1; RESPONSES; VIREMIA C1 Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. NIAID, NIH, Bethesda, MD 20892 USA. RP Blankson, JN (reprint author), Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. NR 6 TC 9 Z9 9 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD JUL 10 PY 2002 VL 288 IS 2 BP 162 EP 164 DI 10.1001/jama.288.2.162-a PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 571HC UT WOS:000176711100013 PM 12095377 ER PT J AU Dorenbaum, A Cunningham, CK Gelber, RD Culnane, M Mofenson, L Britto, P Rekacewicz, C Newell, ML Delfraissy, JF Cunningham-Schrader, B Mirochnick, M Sullivan, JL AF Dorenbaum, A Cunningham, CK Gelber, RD Culnane, M Mofenson, L Britto, P Rekacewicz, C Newell, ML Delfraissy, JF Cunningham-Schrader, B Mirochnick, M Sullivan, JL CA Int PACTG 316 Team TI Two-dose intrapartum/newborn nevirapine and standard antiretroviral therapy to reduce perinatal HIV transmission - A randomized trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; MATERNAL-INFANT TRANSMISSION; QUALITY-ASSURANCE PROGRAM; VERTICAL TRANSMISSION; PREGNANT-WOMEN; INFECTED WOMEN; ZIDOVUDINE; PREVENTION; TYPE-1; COHORT AB Context A 2-dose intrapartum/newborn nevirapine regimen reduced perinatal human immunodeficiency virus (HIV) transmission in Ugandan women not receiving antenatal antiretroviral therapy (ART). However, it is unknown whether the addition of the 2-dose nevirapine regimen to standard ART would further reduce perinatal HIV transmission. Objective To determine whether a 2-dose nevirapine regimen can decrease perinatal transmission of HIV in nonbreastfeeding women receiving standard ART. Design and Setting International, blinded, placebo-controlled, phase 3 trial enrolling women between May 1997 and June 2000 at clinical sites providing care for HIV infection throughout the United States, Europe, Brazil, and the Bahamas. Participants A total of 1270 women received nevirapine (n=642) or placebo (n=628). Infants were followed up for 6 months to determine HIV-infection status, which was available for 1248 deliveries. Intervention A 200-mg dose of oral nevirapine to women after onset of labor and a 2-mg/kg,dose of oral nevirapine to newborns between 48 and 72 hours after birth., Main Outcome Measures Detection of HIV infection in infants and grade 3 and 4 toxic effects in women and newborns. Results After review by the data and safety monitoring board, the trial was stopped early because the overall transmission rates were significantly lower than assumed for the study design. Antenatal ART included zidovudine alone in 23%; combinations without protease inhibitors in 36%; and combinations with protease inhibitors in 41%. Thirty-four percent of women had elective cesarean delivery. No significant safety concerns were identified for women or infants. Detection of HIV infection occurred in 9 (1.4%; 95% confidence interval (01, 0.6%-2.7%) of 631 nevirapine group deliveries and 10 (1.6%; 95% Cl, 0.8%-2.9%) of 617 placebo group deliveries. The 95% Cl for the difference in transmission rate (-0.2) between the 2 study arms ranged from -1.5% in favor of nevirapine to 1.2% in favor of placebo (P=.82, Fisher exact test). The transmission rate was higher in women with lower baseline CD4 cell counts and higher delivery HIV RNA levels, but there was no significant difference between treatment arms in any subgroup. Conclusion Risk of. perinatal HIV transmission was low and no benefit from additional intrapartum/newborn nevirapine was demonstrated when women received prenatal care and antenatal ART, and elective cesarean section was made available. C1 SUNY Upstate Med Univ, Dept Pediat, Syracuse, NY 13210 USA. Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94143 USA. Harvard Univ, Sch Publ Hlth, Stat & Data Anal Ctr, Boston, MA 02115 USA. NIAID, Rockville, MD USA. NICHHD, Bethesda, MD 20892 USA. Agence Natl Rech SIDA, Villejuif, France. INSERM, Villejuif, France. Inst Child Hlth, Ctr Paediat Epidemiol & Biostat, London, England. CHU Bicetre, Enquete Perinatale Francaise, Paris, France. Frontier Sci, Buffalo, NY USA. Boston Univ, Sch Med, Dept Pediat, Boston, MA 02118 USA. Univ Massachusetts, Sch Med, Dept Pediat & Mol Med, Worcester, MA USA. RP Cunningham, CK (reprint author), SUNY Upstate Med Univ, Dept Pediat, 750 E Adams St, Syracuse, NY 13210 USA. EM cunningc@upstate.edu OI Mofenson, Lynne/0000-0002-2818-9808; Newell, Marie-Louise/0000-0002-1074-7699 NR 26 TC 213 Z9 223 U1 0 U2 7 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60654-0946 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD JUL 10 PY 2002 VL 288 IS 2 BP 189 EP 198 DI 10.1001/jama.288.2.189 PG 10 WC Medicine, General & Internal SC General & Internal Medicine GA 571HC UT WOS:000176711100019 PM 12095383 ER PT J AU Demehin, AA Abugo, OO Jayakumar, R Lakowicz, JR Rifkind, JM AF Demehin, AA Abugo, OO Jayakumar, R Lakowicz, JR Rifkind, JM TI Binding of hemoglobin to red cell membranes with eosin-5-maleimide-labeled band 3: Analysis of centrifugation and fluorescence lifetime data SO BIOCHEMISTRY LA English DT Article ID HUMAN-ERYTHROCYTE-MEMBRANE; REVERSIBLE CONFORMATIONAL EQUILIBRIUM; CYTOPLASMIC DOMAIN; NITROBLUE TETRAZOLIUM; PHYSIOLOGICAL PH; IONIC-STRENGTH; ANION-EXCHANGE; BLOOD-CELLS; PROTEIN; SITES AB We have studied the binding of hemoglobin to the red cell membrane by centrifugation and fluorescence methods. The intact red cell was labeled with eosin-5-maleimide (EM), which specifically reacts with lysine 430 of band 3. Even though this residue is not part of the cytoplasmic domain of band 3 (cdb3) associated with hemoglobin binding. fluorescence quenching was observed when hemoglobin bound to inside-out vesicles (IOVs). The use of fluorescence quenching to measure band 3 binding was quantitatively compared with the binding determined by centrifugation, which measures binding to band 3 and non-band 3 sites. For the centrifugation it was necessary to include the non-band 3 association constants determined from chymotrypsin-treated IOVs. The binding of hemoglobin to band 3 was interpreted in terms of the binding of two hemoglobin tetramers to each band 3 dimer. An anticooperative interaction associated with the conformational change produced when hemoglobin binds results in a 2.8-fold decrease in the intrinsic constant of (1.54 +/- 0.25) x 10(7) M-1 for the binding of the second hemoglobin molecule. From the changes in lifetime produced by binding the first and second hemoglobin molecules, it was possible to show that the conformational change associated with binding the second hemoglobin molecule results in a decrease of the heme-eosin distance from 47.90 to 44.78 Angstrom. Reaction of cyanate with the a-amino group of hemoglobin (HbOCN) is shown to produce a very dramatic decrease in the binding of hemoglobin to both the band 3 and non-band 3 sites. The intrinsic constant for binding the first hemoglobin C, molecule to band 3 decreases by a factor of 29 to (5.34 +/- 0.15) x 10(5) M-1. The anticooperative interaction is greater with the intrinsic constant decreasing by a factor of 3.8 for the binding of the second hemoglobin tetramer to band 3. In addition, the nature of the conformational change produced by binding hemoglobin is very different with the second HbOCN increasing the heme-eosin distance to 55.99 Angstrom. The utilization of eosin-5-maleimide-reacted red cell membrane to study hemoglobin binding makes it possible to directly study the binding to band 3. At the same time a sensitive probe of the conformational changes, which occur when hemoglobin binds to band 3. is provided. C1 NIA, Mol Dynam Sect, Baltimore, MD 21224 USA. Univ Maryland, Sch Med, Ctr Fluorescence Spect, Baltimore, MD 21201 USA. Cent Leather Res Inst, Bio Organ Lab, Chennai, India. RP Rifkind, JM (reprint author), NIA, Mol Dynam Sect, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 47 TC 22 Z9 22 U1 1 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 9 PY 2002 VL 41 IS 27 BP 8630 EP 8637 DI 10.1021/bi02007e PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 570TM UT WOS:000176674100012 PM 12093280 ER PT J AU Luu, KX Kanugula, S Pegg, AE Pauly, GT Moschel, RC AF Luu, KX Kanugula, S Pegg, AE Pauly, GT Moschel, RC TI Repair of oligodeoxyribonucleotides by O-6-alkylguanine-DNA alkyltransferase SO BIOCHEMISTRY LA English DT Article ID HUMAN O(6)-ALKYLGUANINE-DNA ALKYLTRANSFERASE; ESCHERICHIA-COLI ADA; DNA-REPAIR; O-6-METHYLGUANINE-DNA METHYLTRANSFERASE; QUADRUPLEX STRUCTURES; O-6-BENZYLGUANINE; OLIGODEOXYNUCLEOTIDES; INACTIVATION; O(6)-BENZYLGUANINE; BINDING AB Activity of the DNA repair protein O-6-alkylguanine-DNA alkyltransferase (AGT) is an important source of tumor cell resistance to alkylating agents. AGT inhibitors may prove useful in enhancing chemotherapy. AGT is inactivated by reacting stoichiometrically with O-6-benzylguanine (b(6)G), which is currently in clinical trials for this purpose. Short oligodeoxyribonucleotides containing a central b(6)G are more potent inactivators of AGT than b(6)G. We examined whether human AGT could react with oligodeoxyribonucleotides containing multiple b(6)G residues. The single-stranded 7-mer 5'-d[T(b(6)G)(5)G]-3' was an excellent AGT substrate with all five b(6)G adducts repaired although one adduct was repaired much more slowly. The highly b(6)G-resistant Y158H and P140K AGT mutants were also inactivated by 5'-d[T(b(6)G)(5)G]-3'. Studies with 7-mers containing a single b(6)G adduct showed that 5'-d[TGGGG(b(6)G)G]-3' was more poorly repaired by wild-type AGT than 5'-d[T(b'G)GGGGG]-3' and 5'-d[TGG(b(6)G)GGG]-3' and was even less repairable by mutants Y158H and P140K. This positional effect was unaffected by interchanging the terminal 5'- or 3'-nucleotides and was also observed with single-stranded 16-mer oligodeoxyribonucleotides containing O-6-Methylguanine, where a minimum of four nucleotides 3' to the lesion was required for the most efficient repair. Annealing with the reverse complementary strands to produce double-stranded substrates increased the ability of AGT to repair adducts at all positions except at positions 2 and 15. Our results suggest that AGT recognizes the polarity of single-stranded DNA, with the best substrates having an adduct adjacent to the 5'-terminal residue. These findings will aid in designing novel AGT inhibitors that incorporate O-6-alkylguanine adducts in oligodeoxyribonucleotide contexts. C1 Penn State Univ, Milton S Hershey Med Ctr, Coll Med, Dept Cellular & Mol Physiol, Hershey, PA 17033 USA. Natl Canc Inst, Comparat Carcinogenesis Lab, Ft Detrick, MD 21702 USA. RP Pegg, AE (reprint author), Penn State Univ, Milton S Hershey Med Ctr, Coll Med, Dept Cellular & Mol Physiol, POB 850,500 Univ Dr, Hershey, PA 17033 USA. EM apegg@psu.edu FU NCI NIH HHS [CA-18137, CA-71976] NR 52 TC 20 Z9 20 U1 0 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 9 PY 2002 VL 41 IS 27 BP 8689 EP 8697 DI 10.1021/bi025857i PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 570TM UT WOS:000176674100019 PM 12093287 ER PT J AU Li, Y Gong, YC Shi, GB Blaszczyk, J Ji, XH Yan, HG AF Li, Y Gong, YC Shi, GB Blaszczyk, J Ji, XH Yan, HG TI Chemical transformation is not rate-limiting in the reaction catalyzed by Escherichia coli 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase SO BIOCHEMISTRY LA English DT Article ID CRYSTAL-STRUCTURE; TERNARY COMPLEX; BINDING; EXPRESSION; RESISTANCE; CLONING; ANALOG; GENE AB 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes the transfer of pyrophosphate from ATP to 6-hydroxymethyl-7,8-dihydropterin (HMDP). Because HPPK is essential for microorganisms but is absent from human and animals, the enzyme is an excellent target for developing antimicrobial agent. Thermodynamic analysis shows that Mg2+ is important not only for the binding of nucleotides but also for the binding of HMDP. Transient kinetic analysis shows that a step or steps after the chemical transformation are rate-limiting in the reaction catalyzed by HPPK. The pre-steady-state kinetics is composed of a burst phase and a steady-state phase. The rate constant for the burst phase is similar to50 times larger than that for the steady-state phase. The latter is very similar to the k(cat) value measured by steady-state kinetics. A set of rate constants for the individual steps of the HPPK-catalyzed reaction has been determined by a combination of stopped-flow and quench-flow analyses. These results form a thermodynamic and kinetic framework for dissecting the roles of active site residues in the substrate binding and catalysis by HPPK. C1 Michigan State Univ, Dept Biochem & Mol Biol, E Lansing, MI 48824 USA. Natl Canc Inst, Macromol Crystallog Lab, Ft Detrick, MD 21702 USA. RP Yan, HG (reprint author), Michigan State Univ, Dept Biochem & Mol Biol, E Lansing, MI 48824 USA. RI Ji, Xinhua/C-9664-2012 OI Ji, Xinhua/0000-0001-6942-1514 FU NIGMS NIH HHS [GM51901] NR 25 TC 31 Z9 31 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD JUL 9 PY 2002 VL 41 IS 27 BP 8777 EP 8783 DI 10.1021/bi025968h PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 570TM UT WOS:000176674100029 PM 12093297 ER PT J AU Lenfant, C AF Lenfant, C TI The SPARK that ignited a new era of research SO CIRCULATION LA English DT Editorial Material DE editorials; genetics; cardiovascular diseases C1 NHLBI, Dept Hlth & Human Serv, NIH, Bethesda, MD 20892 USA. RP Lenfant, C (reprint author), NHLBI, Dept Hlth & Human Serv, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 0 TC 3 Z9 3 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUL 9 PY 2002 VL 106 IS 2 BP 162 EP 163 DI 10.1161/01.CIR.0000025219.04431.E7 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 573GG UT WOS:000176820500010 PM 12105149 ER PT J AU Prasad, A Zhu, JH Halcox, JPJ Waclawiw, MA Epstein, SE Quyyumi, AA AF Prasad, A Zhu, JH Halcox, JPJ Waclawiw, MA Epstein, SE Quyyumi, AA TI Predisposition to atherosclerosis by infections role of endothelial dysfunction SO CIRCULATION LA English DT Article DE atherosclerosis; infection; viruses; endothelium; coronary disease ID CORONARY-ARTERY DISEASE; HERPES-SIMPLEX VIRUS; FUTURE MYOCARDIAL-INFARCTION; NITRIC-OXIDE ACTIVITY; C-REACTIVE PROTEIN; CHLAMYDIA-PNEUMONIAE; HEART-DISEASE; CYTOMEGALOVIRUS-INFECTION; HELICOBACTER-PYLORI; CARDIOVASCULAR-DISEASE AB Background-Several microorganisms have been implicated in the pathogenesis of atherosclerosis. We hypothesized that infections may predispose to atherosclerosis by inflicting endothelial injury. Methods and Results-Of 375 patients undergoing coronary angiography, 218 had assessment of endothelial function using intracoronary acetylcholine (ACH) and of endothelium-independent function with sodium nitroprusside and adenosine. Immunoglobulin-G antibody titers to cytomegalovirus, Chlamydia pneumoniae, Helicobacter pylori, hepatitis A virus, and herpes simplex virus-1 were measured. Pathogen burden was defined as the number of positive antibodies. Although positive serology to individual pathogens tended to be associated with increased incidence of coronary arteriosclerosis (CAD), the pathogen burden correlated with the presence of CAD, even after adjustment for risk factors (OR 1.3; 95% Cl, 1.05 to 1.6, P=0.018). Moreover, the severity of CAD was independently associated with the pathogen burden (P=0.001). Pathogen burden was an independent predictor of endothelial dysfunction, determined as the percent change in coronary vascular resistance in response to ACH (P=0.009) but not the responses to sodium nitroprusside or adenosine. Pathogen burden was also an independent determinant of endothelial function in the subgroup with angiographically normal coronary arteries. Conclusions-The immunoglobulin-G antibody response to multiple pathogens (pathogen burden) is an independent risk factor for endothelial dysfunction and the presence and severity of CAD. Endothelial dysfunction provides the crucial link by which pathogens may contribute to atherogenesis. C1 Washington Hosp Ctr, NHLBI, Cardiovasc Res Inst, Medstar Res Inst, Washington, DC 20010 USA. NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Quyyumi, AA (reprint author), Emory Univ Hosp, 1364 Clifton Rd NE, Atlanta, GA 30322 USA. OI Halcox, Julian/0000-0001-6926-2947 NR 35 TC 197 Z9 212 U1 0 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUL 9 PY 2002 VL 106 IS 2 BP 184 EP 190 DI 10.1161/01.CIR.0000021125.83697.21 PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 573GG UT WOS:000176820500017 PM 12105156 ER PT J AU Oldham, CE Mohney, RP Miller, SLH Hanes, RN O'Bryan, JP AF Oldham, CE Mohney, RP Miller, SLH Hanes, RN O'Bryan, JP TI The ubiquitin-interacting motifs target the endocytic adaptor protein epsin for ubiquitination SO CURRENT BIOLOGY LA English DT Article ID SYNAPTIC GROWTH; ENTH DOMAIN; BINDING; CLATHRIN; DEGRADATION; DROSOPHILA; SITES; EPS15 AB The covalent attachment of ubiquitin to proteins is an evolutionarily conserved signal for rapid protein degradation. However, additional cellular functions for ubiquitination are now emerging, including regulation of protein trafficking and endocytosis [1]. For example, recent genetic studies suggested a role for ubiquitination in regulating epsin, a modular endocytic adaptor protein that functions in the assembly of clathrin-coated vesicles [2, 3]; however, biochemical evidence for this notion has been lacking. Epsin consists of an epsin NH2-terminal homology (ENTH) domain that promotes the interaction with phospholipids [4], several AP2 binding sites [5], two clathrin binding sequences [6], and several Eps15 homology (EH) domain binding motifs [3]. Interestingly, epsin also possesses several recently described ubiquitin-interacting motifs (UIMs) that have been postulated to bind ubiquitin [7]. Here, we demonstrate that epsin is predominantly monoubiquitinated and resistant to proteasomal degradation. The UIMs are necessary for epsin ubiquitination but are not the site of ubiquitination. Finally, we demonstrate that the isolated UIMs from both epsin and an unrelated monoubiquitinated protein, Eps15 [8], are sufficient to promote ubiquitination of a chimeric glutathione-S-transferase (GST)-UIM fusion protein. Thus, our data suggest that UIMs may serve as a general signal for ubiquitination. C1 Natl Inst Environm Hlth Sci, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Biomed Engn, Sch Med, Chapel Hill, NC 27599 USA. RP O'Bryan, JP (reprint author), Natl Inst Environm Hlth Sci, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. NR 23 TC 84 Z9 85 U1 1 U2 4 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE,, CAMBRIDGE, MA 02138 USA SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 9 PY 2002 VL 12 IS 13 BP 1112 EP 1116 AR PII S0960-9822(02)00900-4 DI 10.1016/S0960-9822(02)00900-4 PG 5 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 574YA UT WOS:000176916900021 PM 12121618 ER PT J AU Lovestone, S Hardy, J AF Lovestone, S Hardy, J TI Psychotic genes or forgetful ones? SO NEUROLOGY LA English DT Editorial Material ID ONSET ALZHEIMERS-DISEASE; CHROMOSOME-10; ASSOCIATION; AGGRESSION; SYMPTOMS; 5-HT2A; LOCUS C1 Inst Psychiat, Dept Old Age Psychiat, London SE5 8AF, England. Inst Psychiat, Dept Neurosci, London SE5 8AF, England. NIA, Neurogenet Lab, NIH, Bethesda, MD 20892 USA. RP Lovestone, S (reprint author), Inst Psychiat, Dept Old Age Psychiat, De Crespigny Pk, London SE5 8AF, England. RI Lovestone, Simon/E-8725-2010; Hardy, John/C-2451-2009 NR 14 TC 4 Z9 4 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUL 9 PY 2002 VL 59 IS 1 BP 11 EP 12 PG 2 WC Clinical Neurology SC Neurosciences & Neurology GA 569VG UT WOS:000176622600005 PM 12105301 ER PT J AU Lacourciere, GM Levine, RL Stadtman, TC AF Lacourciere, GM Levine, RL Stadtman, TC TI Direct detection of potential selenium delivery proteins by using an Escherichia coli strain unable to incorporate selenium from selenite into proteins SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID SELD GENE-PRODUCT; TRANSFER-RNA; SELENOCYSTEINE AB Selenium can be metabolized for protein synthesis by two major pathways in vivo. In a specific pathway it can be inserted into polypeptide chains as the amino acid selenocysteine, as directed by the UGA codon. Alternatively, selenium can be substituted for sulfur to generate the free amino acids selenocysteine and selenomethionine, and these are incorporated nonspecifically into proteins in place of cysteine and methionine, respectively. A mutant strain of Escherichia coli was constructed that is deficient in utilization of inorganic selenium for both specific and nonspecific pathways of selenoprotein synthesis. Disruption of the cysK gene prevented synthesis of free cysteine and selenocysteine from inorganic S and Se precursors. Inactivation of the selD gene prevented synthesis of selenophosphate, the reactive selenium donor, required for the specific incorporation pathway. As expected, the double mutant strain, RL165DeltaselD, when grown anaerobically in LB + glucose medium containing (SeO32-)-Se-75, failed to synthesize selenium-dependent formate dehydrogenase H and seleno-tRNAs. However, it incorporated 24% as much selenium as the wild-type strain. Selenium in the deficient strain was bound to five different proteins. A 39-kDa species was identified as glyceraldehyde-3-phosphate dehydrogenase. It is possible that selenium was bound as a perselenide derivative to the reactive cysteine residue of this enzyme. A 28-kDa protein identified as deoxyribose phosphate aldolase also contained bound selenium. These Se-75-labeled proteins may have alternate roles as selenium delivery proteins. C1 NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Stadtman, TC (reprint author), NHLBI, Biochem Lab, NIH, Bldg 50,Room 2120,50 South Dr,MSC-8012, Bethesda, MD 20892 USA. RI Levine, Rodney/D-9885-2011 NR 18 TC 36 Z9 39 U1 0 U2 6 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9150 EP 9153 DI 10.1073/pnas.142291199 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400016 PM 12084818 ER PT J AU Yang, HT Tweedie, D Wang, S Guia, A Vinogradova, T Bogdanov, K Allen, PD Stern, MD Lakatta, EG Boheler, KR AF Yang, HT Tweedie, D Wang, S Guia, A Vinogradova, T Bogdanov, K Allen, PD Stern, MD Lakatta, EG Boheler, KR TI The ryanodine receptor modulates the spontaneous beating rate of cardiomyocytes during development SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID EMBRYONIC STEM-CELLS; SODIUM-CALCIUM EXCHANGER; CARDIAC-SPECIFIC GENES; SIMPLEX VIRUS TYPE-1; IN-VITRO; HEART; CHANNELS; MYOCYTES; CARDIOGENESIS; RELEASE AB in adult myocardium, the heartbeat originates from the sequential activation of ionic currents in pacemaker cells of the sinoatrial node. Ca2+ release via the ryanodine receptor (RyR) modulates the rate at which these cells beat. In contrast, the mechanisms that regulate heart rate during early cardiac development are poorly understood. Embryonic stem (ES) cells can differentiate into spontaneously contracting myocytes whose beating rate increases with differentiation time. These cells thus offer an opportunity to determine the mechanisms that regulate heart rate during development. Here we show that the increase in heart rate with differentiation is markedly depressed in ES cell-derived cardiomyocytes with a functional knockout (KO) of the cardiac ryanodine receptor (RyR2). KO myocytes show a slowing of the rate of spontaneous diastolic depolarization and an absence of calcium sparks. The depressed rate of pacemaker potential can be mimicked in wild-type myocytes by ryanodine, and rescued in KO myocytes with herpes simplex virus (HSV)-1 amplicons containing full-length RyR2. We conclude that a functional RyR2 is crucial to the progressive increase in heart rate during differentiation of ES cell-derived cardiomyocytes, consistent with a mechanism that couples Ca2+ release via RyR before an action potential with activation of an inward current that accelerates membrane depolarization. C1 NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Chinese Acad Sci, Ctr Hlth Sci, Shanghai Inst Biol Sci, Shanghai 200025, Peoples R China. Brigham & Womens Hosp, Dept Anesthesia, Boston, MA 02115 USA. RP Boheler, KR (reprint author), NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 29 TC 70 Z9 79 U1 1 U2 9 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9225 EP 9230 DI 10.1073/hpnas.142651999 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400029 PM 12089338 ER PT J AU Kostrouchova, M Housa, D Kostrouch, Z Saudek, V Rall, JE AF Kostrouchova, M Housa, D Kostrouch, Z Saudek, V Rall, JE TI SKIP is an indispensable factor for Caenorhabditis elegans development SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE transcription; nuclear hormone receptors; cofactors; molting ID THYROID-HORMONE RECEPTOR; RNA-POLYMERASE-II; GATA-FACTOR; TRANSCRIPTIONAL REPRESSION; DROSOPHILA-MELANOGASTER; PROTEIN INTERACTS; HOMOLOG; COMPLEX; GENE; REDISTRIBUTION AB SKI-binding protein (SKIP) is a transcription cofactor present in all eukaryotes. Here we show that SKIP is a unique protein that is required for Caenorhabditis elegans viability and development. Expression of CeSKIP (skp-1) assayed by RT-PCR and by GFP fluorescence in transgenic lines starts in embryos and continues to adulthood. Loss of CeSKIP activity by RNA-mediated inhibition results in early embryonic arrest similar to that seen following inhibition of RNA polymerase Ill. RNA polymerase 11 phosphorylation appears normal early in CeSKIP RNA-mediated inhibition treated embryos although the expression of several embryonic GFP reporter genes is severely restricted or absent. Our data suggest that CeSKIP is an essential component of many RNA polymerase 11 transcription complexes and is indispensable for C. elegans development. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. Incyte Genom, Cambridge CB2 1FF, England. Charles Univ Prague, Lab Biol & Genet, Prague 12808 2, Czech Republic. Charles Univ Prague, Lab Mol Pathol, Inst Inherited Metab Disorders, Fac Med 1, Prague 12808 2, Czech Republic. RP Rall, JE (reprint author), NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. EM joseph.rall@intra.niddk.nih.gov RI Housa, Daniel/B-2370-2008 NR 42 TC 24 Z9 29 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9254 EP 9259 DI 10.1073/pnas.112213799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400034 PM 12084813 ER PT J AU Cicala, C Arthos, J Selig, SM Dennis, G Hosack, DA Van Ryk, D Spangler, ML Steenbeke, TD Khazanie, P Gupta, N Yang, J Daucher, M Lempicki, RA Fauci, AS AF Cicala, C Arthos, J Selig, SM Dennis, G Hosack, DA Van Ryk, D Spangler, ML Steenbeke, TD Khazanie, P Gupta, N Yang, J Daucher, M Lempicki, RA Fauci, AS TI HIV envelope induces a cascade of cell signals in non-proliferating target cells that favor virus replication SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; CD4(+) T-CELLS; FOCAL ADHESION KINASE; MACROPHAGE-TROPIC HIV; CHEMOKINE RECEPTOR; MEMBRANE-FUSION; GENE-EXPRESSION; BETA-CHEMOKINES; NUCLEAR FACTOR; INFECTION AB Certain HIV-encoded proteins modify host-cell gene expression in a manner that facilitates viral replication. These activities may contribute to low-level viral replication in nonproliferating cells. Through the use of oligonucleotide microarrays and high-throughput Western blotting we demonstrate that one of these proteins, gp120, induces the expression of cytokines, chemokines, kinases, and transcription factors associated with antigen-specific T cell activation in the absence of cellular proliferation. Examination of transcriptional changes induced by gp120 in freshly isolated peripheral blood mononuclear cells and monocyte-derived macrophages reveals a broad and complex transcriptional program conducive to productive infection with HIV. Observations include the induction of nuclear factor of activated T cells, components of the RNA polymerase 11 complex including TFII D, proteins localized to the plasma membrane, including several syntaxins, and members of the Rho protein family, including Cdc 42. These observations provide evidence that envelope-mediated signaling contributes to the productive infection of HIV in suboptimally activated T cells. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIAID, Lab Immunopathogenesis & Bioinformat, NIH, Frederick, MD 21702 USA. RP Cicala, C (reprint author), 10 Ctr Dr,MSC 1876,Bldg 10,Room 11B13, Bethesda, MD 20892 USA. RI Lempicki, Richard/E-1844-2012 OI Lempicki, Richard/0000-0002-7059-409X NR 45 TC 123 Z9 126 U1 0 U2 3 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9380 EP 9385 DI 10.1073/pnas.142287999 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400056 PM 12089333 ER PT J AU Davis, VL Chan, CC Schoen, TJ Couse, JF Chader, GJ Korach, KS AF Davis, VL Chan, CC Schoen, TJ Couse, JF Chader, GJ Korach, KS TI An estrogen receptor repressor induces cataract formation in transgenic mice SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID AGE; EYE; PREVENTION; EXPRESSION; TAMOXIFEN; THERAPY; BETA AB Despite the high prevalence of age-related cataracts, there are currently no known therapies to delay or prevent their occurrence. Studies in humans and rodent models suggest that estrogen may provide protection against age-related cataracts. The discovery of ocular estrogen receptors (ERs) indicates that estrogen protection may result from direct interactions with its receptors in the eye, instead an indirect consequence from effects on another tissue. Studies in our transgenic mouse model validate the concept that estrogen is beneficial for the eye. These mice express ERDelta3, a dominant-negative form of ERalpha that inhibits ERalpha function. In the ERDelta3 transgenic mice, cortical cataracts spontaneously form in ERDelta3 females after puberty and progress with age. The cataracts initiate in the equatorial region of the lens where the epithelial cells differentiate into elongating fiber cells. Cataract formation can be prevented if the females are ovariectomized before, but not after, sexual maturity. Both male and female ERDelta3 mice develop cataracts after neonatal treatment with the potent estrogen diethyl ethylstilbestrol (DES). The incidence of spontaneous and DES-induced cataracts in ERDelta3 mice is 100%, yet these cataracts are absent from the wild-type mice. These data suggest that repression of estrogen action induces cortical cataract formation because estrogen is required to activate the ERDelta3 repressor. Evidence of DES-induced cataracts in the ERDelta3 males as well as the females suggests that estrogen is important in lens physiology in both sexes. The ERDelta3 mice provide a powerful model for assessing the role of estrogen in maintaining the transparency of the lens. C1 NIEHS, Receptor Biol Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Davis, VL (reprint author), Cedars Sinai Med Ctr, Burns & Allen Res Inst, 3005 Davis Bldg,8700 Beverly Blvd, Los Angeles, CA 90048 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 22 TC 23 Z9 27 U1 0 U2 3 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9427 EP 9432 DI 10.1073/pnas.132247999 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400064 PM 12082175 ER PT J AU Julias, JG McWilliams, MJ Sarafianos, SG Arnold, E Hughes, SH AF Julias, JG McWilliams, MJ Sarafianos, SG Arnold, E Hughes, SH TI Mutations in the RNase H domain of HIV-1 reverse transcriptase affect the initiation of DNA synthesis and the specificity of RNase H cleavage in vivo SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; MURINE LEUKEMIA-VIRUS; DOUBLE-STRANDED DNA; RIBONUCLEASE-H; ANGSTROM RESOLUTION; CRYSTAL-STRUCTURE; CIRCLE JUNCTION; TYPE-1; PRIMER; REPLICATION AB Retroviral reverse transcriptases contain a DNA polymerase activity that can copy an RNA or DNA template and an RNase H activity that degrades the viral RNA genome during reverse transcription. RNase H makes both specific and nonspecific cleavages; specific cleavages are used to generate and remove the polypurine tract primer used for plus-strand DNA synthesis and to remove the tRNA primer used for minus-strand DNA synthesis. We generated mutations in an HIV-1-based vector to change amino acids in the RNase H domain that contact either the RNA and DNA strands. Some of these mutations affected the initiation of DNA synthesis, demonstrating an interdependence of the polymerase and RNase H activities of HIV-1 reverse transcription during viral DNA synthesis. The ends of the linear DNA form of the HIV-1 genome are defined by the specific RNase H cleavages that remove the plus- and minus-strand primers; these ends can be joined to form two-long-terminal repeat circles. Analysis of two-long-terminal repeat circle junctions showed that mutations in the RNase H domain affect the specificity of RNase H cleavage. C1 NCI, HIV Drug Resistance Program, Frederick, MD 21702 USA. Rutgers State Univ, Ctr Adv Biotechnol & Med, Piscataway, NJ 08854 USA. Rutgers State Univ, Dept Chem, Piscataway, NJ 08854 USA. RP Hughes, SH (reprint author), NCI, HIV Drug Resistance Program, POB B, Frederick, MD 21702 USA. EM hughes@ncifcrf.gov OI Sarafianos, Stefan G/0000-0002-5840-154X FU NIAID NIH HHS [AI 09578, AI 27690, F32 AI009578]; NIGMS NIH HHS [GM55609] NR 27 TC 74 Z9 75 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9515 EP 9520 DI 10.1073/pnas.142123199 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400079 PM 12093908 ER PT J AU Tomac, AC Agulnick, AD Haughey, N Chang, CF Zhang, YJ Backman, C Morales, M Mattson, MP Wang, Y Westphal, H Hoffer, BJ AF Tomac, AC Agulnick, AD Haughey, N Chang, CF Zhang, YJ Backman, C Morales, M Mattson, MP Wang, Y Westphal, H Hoffer, BJ TI Effects of cerebral ischemia in mice deficient in Persephin SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NEUROTROPHIC FACTOR; NERVOUS-SYSTEM; LACKING GDNF; DOPAMINE NEURONS; IN-VIVO; RECEPTOR; FAMILY; NEURTURIN; EXPRESSION; PROTECTION AB Persephin (Pspn), a recently cloned member of the transforming growth factor-beta superfamily (TGF-beta) and glial cell line-derived neurotrophic factor (GDNF) subfamily, is distributed throughout the nervous system at extremely low levels and is thought to function as a survival factor for midbrain dopaminergic and spinal motor neurons in vivo. Here, we report that mice lacking Pspn by homologous recombination show normal development and behavior, but are hypersensitive to cerebral ischemia. A 300% increase in infarction volume was observed after middle cerebral artery occlusion. We find that glutamate-induced Ca2+ influx, thought to be a major component of ischemic neuronal cell death, can be regulated directly by the Persephin protein (PSP) and that PSP can reduce hypoxia/reperfusion cell death in vitro. Neuronal cell death can be prevented or markedly attenuated by administration of recombinant human PSP in vivo before ischemia in both mouse and rat models. Taken together, these data indicate that PSP is a potent modulator of excitotoxicity in the central nervous system with pronounced neuroprotective activity. Our findings support the view that PSP signaling can exert an important control function in the context of stroke and glutamate-mediated neurotoxicity, and also suggest that future therapeutic approaches may involve this novel trophic protein. C1 NIDA, NIH, Baltimore, MD 21224 USA. NICHD, NIH, Bethesda, MD 20892 USA. NIA, NIH, Bethesda, MD 20892 USA. Tri Serv Gen Hosp, Natl Def Med Ctr, Taipei, Taiwan. RP Hoffer, BJ (reprint author), NIDA, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012; backman, cristina/C-1276-2013 NR 40 TC 44 Z9 45 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUL 9 PY 2002 VL 99 IS 14 BP 9521 EP 9526 DI 10.1073/pnas.152535899 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 572LE UT WOS:000176775400080 PM 12093930 ER EF