FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Lakatta, EG AF Lakatta, EG TI Cardiovascular aging in health SO CLINICS IN GERIATRIC MEDICINE LA English DT Article ID AGE-ASSOCIATED CHANGES; SARCOPLASMIC-RETICULUM CA-2+-ATPASE; RAT MYOCARDIUM; HUMAN HEART; EXCITATION-CONTRACTION; DYNAMIC EXERCISE; ATPASE ACTIVITY; BLOOD-PRESSURE; MESSENGER-RNA; YOUNG-ADULT AB Major focuses of geriatric medicine are cardiovascular diseases with increases in prevalence in older people and how aging might modify the risk factors, clinical manifestations, treatment strategies, and prognoses of these diseases. Quantitative information on age-associated alterations in cardiovascular structure and function in health is essential to define and target the specific characteristics of the cardiovascular aging process that render it the major risk factor for cardiovascular diseases. Such information also is required to differentiate among the limitations of elderly individuals that relate to disease and limitations that may fall within expected normal limits. Differences in cardiovascular function between older and younger individuals have been described extensively in the literature, but confusion often arises in the interpretation of these differences because of a failure to acknowledge or to control for interactions among age, disease, and lifestyle. The Baltimore Longitudinal Study on Aging has made an effort to characterize the effects of aging on multiple aspects of cardiovascular structure and function. The concepts that have evolved from this and other studies are discussed. C1 NIA, Gerontol Res Ctr, Intramural Res Program, Lab Cardiovasc Sci,NIH, Baltimore, MD 21224 USA. RP Lakatta, EG (reprint author), NIA, Gerontol Res Ctr, Intramural Res Program, Lab Cardiovasc Sci,NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 50 TC 54 Z9 58 U1 0 U2 5 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0749-0690 J9 CLIN GERIATR MED JI Clin. Geriatr. Med. PD AUG PY 2000 VL 16 IS 3 BP 419 EP + DI 10.1016/S0749-0690(05)70021-5 PG 27 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 349XJ UT WOS:000089070600003 PM 10918640 ER PT J AU Hall, LL Purdy, R AF Hall, LL Purdy, R TI Recovery and serious brain disorders: The central role of families in nurturing roots and wings SO COMMUNITY MENTAL HEALTH JOURNAL LA English DT Article ID SEVERE MENTAL-ILLNESS; SCHIZOPHRENIA; MEDICATION; BURDEN; INTERVENTION; PERCEPTIONS; ADJUSTMENT; VIOLENCE; SIBLINGS; CONTEXT AB Recovery in severe mental illness is a powerful concept for consumers, invoking a journey through understanding and acceptance of illness and disability, along with hope for a quality life, self-empowerment, and responsibility. Although not widely recognized, the journey toward recovery is also important for family members of individuals with severe mental illnesses, who often serve in a care-giving or supportive role for their loved ones. From the family perspective, to make recovery a real possibility, several issues must be confronted. First, progress toward recovery must acknowledge and involve care-giving and supportive family members, recognizing their significant role in the lives of many individuals with severe mental illnesses as well as the journey of family members in understanding and accepting a severe mental illness in a loved one. Second, a real, recovery-oriented system implements treatments and supports shown to be effective. In this day and age, the gap between research findings and ordinary care condemns too many consumers and their families to outcomes far short of what is possible. This includes not only medications, psychotherapy, employment services, dual diagnosis services, housing and other supports for consumers, but also family education, shown to be effective by a large body of research. Third, a recovery-oriented system of care cannot afford to dance around the thorniest and most controversial issues in severe mental illnesses-such as individuals with the most intractable forms of illness or the relatively infrequent but very real situations in which either consumers or family members are assaultive or abusive. All of these elements are essential if recovery is to be more than a slogan, but rather a true goal in a system of care that respects consumers with these disorders and their care-giving family members. C1 NIMH, Bethesda, MD 20892 USA. RP Hall, LL (reprint author), 6001 Execut Blvd,Suite 8235, Bethesda, MD 20892 USA. NR 33 TC 10 Z9 10 U1 0 U2 1 PU KLUWER ACADEMIC-HUMAN SCIENCES PRESS PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013-1578 USA SN 0010-3853 J9 COMMUNITY MENT HLT J JI Community Ment. Health J. PD AUG PY 2000 VL 36 IS 4 BP 427 EP 441 DI 10.1023/A:1001917114105 PG 15 WC Health Policy & Services; Public, Environmental & Occupational Health; Psychiatry SC Health Care Sciences & Services; Public, Environmental & Occupational Health; Psychiatry GA 330NC UT WOS:000087967300008 PM 10917276 ER PT J AU Francischetti, IMB Gombarovits, MEC Valenzuela, JG Carlini, CR Guimaraes, JA AF Francischetti, IMB Gombarovits, MEC Valenzuela, JG Carlini, CR Guimaraes, JA TI Intraspecific variation in the venoms of the South American rattlesnake (Crotalus durissus terrificus) SO COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY LA English DT Article DE Crotalus durissus terrificus; South American rattlesnake; intraspecific variation; Minas Gerais State; crotoxin; convulxin; crotamine; isoforms ID THROMBIN-LIKE ENZYME; NOTECHIS-ATER-NIGER; SNAKE-VENOMS; SCUTULATUS-SCUTULATUS; BIOLOGICAL-ACTIVITIES; BOTHROPS; POPULATIONS; PROTEINS; A(2); CHROMATOGRAPHY AB The venom of eight individual Crotalus durissus terrificus snakes from the State of Minas Gerais, Brazil, in addition to pooled venom from Butantan Institute, were compared. Snakes were captured in distinct locations, some of them 600 km apart: Conselheiro Lafaiete, Entre Rios de Minas, Itauna, Itapecerica, Lavras, Patos de Minas, Paracatu, and Santo Antonio do Amparo. The crude venoms were tested for proteolytic, phospholipase A2, platelet aggregating, and hemagglutinating activities. The venoms were also analyzed by polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing (IEF). Chromatographic patterns of venom proteins on both gel-filtration and anion-exchange chromatographies were also performed. All venoms presented high phospholipase A2 and platelet-aggregating activities, but only minimal hemagglutinating or proteolytic activities were found. Gel-filtration chromatography showed a characteristic profile for most venoms where four main peaks were separated, including the typical ones where convulxin and crotoxin were identified; however, peaks with high amounts of lower molecular weight proteins were found in the venoms from the Santo Antonio do Amparo location and Butantan Institute, characterizing these venoms as crotamine positive. Anion-exchange chromatographies presented a similar protein distribution pattern, although the number of peaks (up to ten) distinguished some Venom samples. Consistent with these results, polyacrylamide gels that were silver stained after venom separation by PAGE or IEF presented a similar qualitative band distribution, although a quantitative heterogeneity was detected among venoms. Our results suggest that the variability found in venom components of C. d. terrificus venoms captured in Minas Gerais State may be genetically inherited and/or environmentally induced. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Fed Rio Grande Sul, Ctr Biotecnol, BR-91501970 Porto Alegre, RS, Brazil. Fed Univ Rio De Janeiro, CCS, ICB, Dept Biochem Med, BR-21941590 Rio De Janeiro, Brazil. NIAID, Parasit Dis Lab, Med Entomol Sect, NIH, Bethesda, MD 20892 USA. RP Guimaraes, JA (reprint author), Univ Fed Rio Grande Sul, Ctr Biotecnol, Av Bento Goncalves 9500,CP 15005, BR-91501970 Porto Alegre, RS, Brazil. OI Guimaraes, Jorge A./0000-0001-6354-6789; Carlini, Celia R/0000-0002-7027-2445 NR 48 TC 28 Z9 30 U1 0 U2 8 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1532-0456 J9 COMP BIOCHEM PHYS C JI Comp. Biochem. Physiol. C-Pharmacol. Toxicol. Endocrinol. PD AUG PY 2000 VL 127 IS 1 BP 23 EP 36 DI 10.1016/S0742-8413(00)00129-8 PG 14 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Toxicology; Zoology SC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Toxicology; Zoology GA 398TT UT WOS:000166773100003 PM 11081410 ER PT J AU Strandberg, JD Whitehair, LA AF Strandberg, JD Whitehair, LA TI National Institutes of Health - Supported opportunities in comparative medicine SO COMPARATIVE MEDICINE LA English DT Article AB The Comparative Medicine (CM) area of the National Institutes of Health (NIH) is a major source of support for research on laboratory animals, training of laboratory animal specialists, and support of shared, regional animal resources. We present a brief history of CM at NIH and the major mechanisms by which it accomplishes its goals in programs located across the United States. C1 NIH, Natl Ctr Res Resources, Comparat Med Area, Bethesda, MD 20892 USA. RP Strandberg, JD (reprint author), NIH, Natl Ctr Res Resources, Comparat Med Area, 6705 Rockledge Dr, Bethesda, MD 20892 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 0023-6764 J9 COMPARATIVE MED JI Comparative Med. PD AUG PY 2000 VL 50 IS 4 BP 356 EP 362 PG 7 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 354MH UT WOS:000089333900003 PM 11020151 ER PT J AU Marcin, JP Pollack, MM Patel, KM Ruttimann, UE AF Marcin, JP Pollack, MM Patel, KM Ruttimann, UE TI Combining physician's subjective and physiology-based objective mortality risk predictions SO CRITICAL CARE MEDICINE LA English DT Article DE critical care; intensive care; pediatrics; decision-making; Bayes Theorem; mortality; Pediatric Risk of Mortality III; severity of illness ID INTENSIVE-CARE UNITS; ILL HOSPITALIZED ADULTS; CLINICAL JUDGMENT; INCEPTION COHORT; SCORING SYSTEMS; APACHE-II; MULTICENTER; MODELS; MANAGEMENT; PERFORMANCE AB Objective: None of the currently available physiology-based mortality risk prediction models incorporate subjective judgements of healthcare professionals, a source of additional information that could improve predictor performance and make such systems more acceptable to healthcare professionals. This study compared the performance of subjective mortality estimates by physicians and nurses with a physiology-based method, the Pediatric Risk of Mortality (PRISM) III, Then, healthcare provider estimates were combined with PRISM III estimates using Bayesian statistics. The performance of the Bayesian model was then compared with the original two predictions. Design: Concurrent cohort study. Setting: A tertiary pediatric intensive care unit at a university affiliated children's hospital. Patients: Consecutive admissions to the pediatric intensive care unit. Interventions: None. Measurements and Main Results: For each of the 642 consecutive eligible patients, an exact mortality estimate and the degree of certainty (continuous scale from 1 to 5) associated with the estimate was collected from the attending, fellow, resident, and nurse responsible for the patient's care. Bayesian statistics were used to combine the PRISM III and certainty weighted subjective predictions to create a third Bayesian estimate of mortality. PRISM III discriminated survivors from nonsurvivors very well (area under curve [AUC], 0.924) as did the physicians and nurses (AUCs attendings, 0.953; fellows, 0.870; residents, 0.923; nurses, 0.935). Although the AUCs of the healthcare providers were not significantly different from the AUCs of PRISM III, the Bayesian AUCs were higher than both the healthcare providers' AUCs (p less than or equal to .09 for all) and PRISM III AUCs. Similarly, the calibration statistics for the Bayesian estimates were superior to the calibration statistics for both the healthcare providers and PRISM III models. Conclusions: The results of this study demonstrated that healthcare providers' subjective mortality predictions and PRISM III mortality predictions perform equally well. The Bayesian model that combined provider and PRISM III mortality predictions was more accurate than either provider or PRISM III alone and may be more acceptable to physicians. A methodology using subjective outcome predictions could be more relevant to individual patient decision support. C1 George Washington Univ, Sch Med,Childrens Natl Med Ctr, Childrens Res Inst, Ctr Hlth Serv & Clin Res,Sect Crit Care Med, Washington, DC 20052 USA. NIAA, NIH, Bethesda, MD USA. NIH, Dept Crit Care Med, Bethesda, MD 20892 USA. Univ Calif Davis, Dept Pediat, Sect Crit Care Med, Davis, CA 95616 USA. RP Marcin, JP (reprint author), George Washington Univ, Sch Med,Childrens Natl Med Ctr, Childrens Res Inst, Ctr Hlth Serv & Clin Res,Sect Crit Care Med, Washington, DC 20052 USA. NR 39 TC 17 Z9 19 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD AUG PY 2000 VL 28 IS 8 BP 2984 EP 2990 DI 10.1097/00003246-200008000-00050 PG 7 WC Critical Care Medicine SC General & Internal Medicine GA 346JM UT WOS:000088867300048 PM 10966283 ER PT J AU Donaldson, JG Jackson, CL AF Donaldson, JG Jackson, CL TI Regulators and effectors of the ARF GTPases SO CURRENT OPINION IN CELL BIOLOGY LA English DT Review ID ADP-RIBOSYLATION FACTOR; NUCLEOTIDE EXCHANGE FACTOR; ACTIVATING PROTEIN; GOLGI-COMPLEX; SEC7 DOMAIN; ACTIN CYTOSKELETON; GTP HYDROLYSIS; KDEL RECEPTOR; BREFELDIN-A; RECRUITMENT AB The small G proteins of the ARF family are key regulators of membrane dynamics. Many functions of ARF proteins in cells are being revealed by studies of their regulators and effecters. Significant progress has been made over the past year, with the identification of a surprisingly large family of novel ARF GTPase-activating proteins. In addition, two new classes of effecters, the PIP kinases and a novel family of monomeric coat-like proteins have been discovered. C1 CEA Saclay, Serv Biochim & Genet Mol, F-91191 Gif Sur Yvette, France. NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Jackson, CL (reprint author), CEA Saclay, Serv Biochim & Genet Mol, Bat 142, F-91191 Gif Sur Yvette, France. RI Jackson, Catherine/A-3421-2013 OI Jackson, Catherine/0000-0002-0843-145X NR 70 TC 297 Z9 302 U1 0 U2 6 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0955-0674 J9 CURR OPIN CELL BIOL JI Curr. Opin. Cell Biol. PD AUG PY 2000 VL 12 IS 4 BP 475 EP 482 DI 10.1016/S0955-0674(00)00119-8 PG 8 WC Cell Biology SC Cell Biology GA 334AH UT WOS:000088162600013 PM 10873831 ER PT J AU Golden, A AF Golden, A TI Cytoplasmic flow and the establishment of polarity in C-elegans 1-cell embryos SO CURRENT OPINION IN GENETICS & DEVELOPMENT LA English DT Review ID ACTIN-ASSOCIATED PROTEINS; CAENORHABDITIS-ELEGANS; CELL-CYCLE; CYTOKINESIS; GENE; EMBRYOGENESIS; GRANULES; DIVISION; MICROFILAMENTS; SPECIFICATION AB Early Caenorhabditis elegans embryos provide an excellent model for the study of developmental processes. Development can be studied by direct observation under the light microscope and can be perturbed using laser manipulations, drug inhibitor treatments, and genetic mutants. The first division of the C. elegans embryo is asymmetric, generating two daughter cells unequal in size and developmental fate. These distinct fates are generated by the partitioning of cytoplasmic determinants during the first mitotic cell cycle. Partitioning of these determinants is thought to be driven by cytoplasmic flow. Recent studies in C, elegans in the past year have identified a number of components necessary for this flow, giving us a clearer picture of the molecular mechanisms underlying developmental asymmetry. C1 NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Golden, A (reprint author), NIDDKD, Lab Biochem & Genet, NIH, 8 Ctr Dr MSCX 0840,Bldg 8,Room 323, Bethesda, MD 20892 USA. NR 39 TC 24 Z9 42 U1 1 U2 2 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 2000 VL 10 IS 4 BP 414 EP 420 DI 10.1016/S0959-437X(00)00106-4 PG 7 WC Cell Biology; Genetics & Heredity SC Cell Biology; Genetics & Heredity GA 336CV UT WOS:000088285100012 PM 10889063 ER PT J AU Gurunathan, S Wu, CY Freidag, BL Seder, RA AF Gurunathan, S Wu, CY Freidag, BL Seder, RA TI DNA vaccines: a key for inducing long-term cellular immunity SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID CYTOTOXIC T-LYMPHOCYTES; CPG-CONTAINING OLIGODEOXYNUCLEOTIDES; INTRADERMAL GENE IMMUNIZATION; ANTIGEN-PRESENTING CELLS; MARROW-DERIVED CELLS; B SURFACE-ANTIGEN; DENDRITIC CELLS; IN-VIVO; PLASMID DNA; IMMUNOSTIMULATORY DNA AB Over the past few years, major advances in several areas of immunology have provided a foundation for the rational design of vaccines against diseases requiring cellular immunity. Among these advances are the cellular mechanisms by which DNA vaccines can sustain long-term humoral and cellular immunity. C1 NIAID, Clin Immunol Sect, Clin Invest Lab, Clin Immunol Sect, Bethesda, MD 20892 USA. RP Gurunathan, S (reprint author), NIAID, Clin Immunol Sect, Clin Invest Lab, Clin Immunol Sect, 10 Ctr Dr,Room 10-11C215, Bethesda, MD 20892 USA. NR 54 TC 164 Z9 187 U1 2 U2 4 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD AUG PY 2000 VL 12 IS 4 BP 442 EP 447 DI 10.1016/S0952-7915(00)00118-7 PG 6 WC Immunology SC Immunology GA 330UD UT WOS:000087978800015 PM 10899026 ER PT J AU Leonard, WJ AF Leonard, WJ TI Genetic effects on immunity - Editorial overview SO CURRENT OPINION IN IMMUNOLOGY LA English DT Editorial Material C1 NHLBI, Lab Mol Immunol, NIH, Bethesda, MD 20892 USA. RP Leonard, WJ (reprint author), NHLBI, Lab Mol Immunol, NIH, Bldg 10,Room 7N252,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 5 TC 1 Z9 1 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD AUG PY 2000 VL 12 IS 4 BP 465 EP 467 DI 10.1016/S0952-7915(00)00121-7 PG 3 WC Immunology SC Immunology GA 330UD UT WOS:000087978800018 ER PT J AU Puel, A Leonard, WJ AF Puel, A Leonard, WJ TI Mutations in the gene for the IL-7 receptor result in T-B+N K+ severe combined immunodeficiency disease SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID B-CELL-DEVELOPMENT; DEFECTIVE LYMPHOID DEVELOPMENT; THYMIC STROMAL LYMPHOPOIETIN; MICE LACKING JAK3; INTERLEUKIN-7 RECEPTOR; GAMMA-CHAIN; DEFICIENT MICE; PHOSPHATIDYLINOSITOL 3-KINASE; PHOSPHOINOSITIDE 3-KINASE; THYMOCYTE MATURATION AB Recently, two SCID (severe combined immunodeficiency disease) patients with greatly diminished T cells but normal or increased numbers of B and NK cells (T-B+NK+SCID) were found to have mutations in the gene for the IL-7 receptor. This has establish ed a major role for IL-7-receptor-dependent signaling in T cell development in humans and probably explains the diminished T cell numbers seen in patients with X-linked SCID or SCID that results from Jak3-deficiency. C1 NHLBI, Lab Mol Immunol, NIH, Bethesda, MD 20892 USA. RP Puel, A (reprint author), NHLBI, Lab Mol Immunol, NIH, Bldg 10,Room 7N252,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 72 TC 49 Z9 51 U1 1 U2 3 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD AUG PY 2000 VL 12 IS 4 BP 468 EP 473 DI 10.1016/S0952-7915(00)00122-9 PG 6 WC Immunology SC Immunology GA 330UD UT WOS:000087978800019 PM 10899029 ER PT J AU Galon, J Aksentijevich, I McDermott, MF O'Shea, JJ Kastner, DL AF Galon, J Aksentijevich, I McDermott, MF O'Shea, JJ Kastner, DL TI TNFRSF1A mutations and autoinflammatory syndromes SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID TUMOR-NECROSIS-FACTOR; FAMILIAL MEDITERRANEAN FEVER; ENCODING MEVALONATE KINASE; FACTOR TNF RECEPTOR; FAS GENE-MUTATIONS; PERIODIC FEVER; LYMPHOPROLIFERATIVE SYNDROME; HYPERIMMUNOGLOBULINEMIA-D; RHEUMATOID-ARTHRITIS; MONOCLONAL-ANTIBODY AB The autoinflammatory syndromes are systemic disorders characterized by apparently unprovoked inflammation in the absence of high-titer autoantibodies or antigen-specific T lymphocytes. One such illness, TN F-receptor-associated periodic syndrome (TRAPS), presents with prolonged attacks of fever and severe localized inflammation. TRAPS is caused by dominantly inherited mutations in TNFRSF1A (formerly termed TNFR1), the gene encoding the 55 kDa TNF receptor. All known mutations affect the first two cysteine-rich extracellular subdomains of the receptor, and several mutations are substitutions directly disrupting conserved disulfide bonds. One likely mechanism of inflammation in TRAPS is the impaired cleavage of TN FRSF1A ectodomain upon cellular activation, with diminished shedding of the potentially antagonistic soluble receptor. Preliminary experience with recombinant p75 TNFR-Fc fusion protein in the treatment of TRAPS has been favorable. C1 NIAMSD, Arthrit & Rheumatism Branch, Bethesda, MD 20892 USA. St Bartholomews & Royal London Hosp, Sch Med & Dent, Med Unit, London E1 1BB, England. RP Galon, J (reprint author), NIAMSD, Arthrit & Rheumatism Branch, 10 Ctr Dr,Room 9N228,MSC 1820, Bethesda, MD 20892 USA. NR 44 TC 172 Z9 184 U1 1 U2 4 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD AUG PY 2000 VL 12 IS 4 BP 479 EP 486 DI 10.1016/S0952-7915(00)00124-2 PG 8 WC Immunology SC Immunology GA 330UD UT WOS:000087978800021 PM 10899034 ER PT J AU McFadden, G Murphy, PM AF McFadden, G Murphy, PM TI Host-related immunomodulators encoded by poxviruses and herpesviruses SO CURRENT OPINION IN MICROBIOLOGY LA English DT Review ID OPEN READING FRAME; CYTOMEGALOVIRUS CHEMOKINE HOMOLOG; COMPLEMENT CONTROL PROTEIN; NECROSIS-FACTOR RECEPTORS; MYXOMA VIRUS; FUNCTIONAL CHEMOKINE; MOLECULAR MIMICRY; BETA-CHEMOKINE; SEQUENCE; HUMAN-HERPESVIRUS-8 AB In the past year, important advances have been made in the area of host-related immunomodulatory genes encoded by the larger DNA viruses, particularly for the poxviruses and herpesviruses. Not only has the repertoire of viral immunomodulator homologs expanded as a result of sequencing the complete genome of another six large DNA viruses, but also new concepts of how they work have been proposed and in some cases supported by in vivo evidence. Recent developments have been made in understanding a spectrum of host-related viral modulators, including complement control proteins, TNF-receptor homologs, IL-18 binding proteins, viral interleukins (vIL-6 and vIL-10), chemokine mimics and chemokine receptor homologs. C1 Univ Western Ontario, John P Robarts Res Inst, London, ON N6G 2V4, Canada. Univ Western Ontario, Dept Microbiol & Immunol, London, ON N6G 2V4, Canada. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP McFadden, G (reprint author), Univ Western Ontario, John P Robarts Res Inst, 1400 Western Rd, London, ON N6G 2V4, Canada. NR 57 TC 80 Z9 86 U1 0 U2 1 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 2000 VL 3 IS 4 BP 371 EP 378 DI 10.1016/S1369-5274(00)00107-7 PG 8 WC Microbiology SC Microbiology GA 345LK UT WOS:000088817300009 PM 10972497 ER PT J AU Friedman, T Battey, J Kachar, B Riazuddin, S Noben-Trauth, K Griffith, A Wilcox, E AF Friedman, T Battey, J Kachar, B Riazuddin, S Noben-Trauth, K Griffith, A Wilcox, E TI Modifier genes of hereditary hearing loss SO CURRENT OPINION IN NEUROBIOLOGY LA English DT Review ID OUTER HAIR-CELLS; DOMINANT DEAFNESS; MUTATIONS; MICE; CONNEXIN-26; LOCUS; IDENTIFICATION; TRANSDUCTION; IMPAIRMENT; TUBBY AB Phenotypic variation between individuals with the same disease alleles may be attributable to the genotype at another locus, which is referred to as a modifier gene. Recent functional studies of modifier genes of hearing-loss loci have begun to refine our understanding of hearing processes and will guide the rational design of medical therapies for hearing loss. C1 Natl Inst Deafness & Other Commun Disorders, Genet Mol Lab, Rockville, MD 20850 USA. Natl Inst Deafness & Other Commun Disorders, Bethesda, MD 20892 USA. Natl Ctr Excellence Mol Biol, Lahore 53700, Pakistan. RP Friedman, T (reprint author), Natl Inst Deafness & Other Commun Disorders, Genet Mol Lab, 5 Res Court, Rockville, MD 20850 USA. FU NIDCD NIH HHS [Z01 DC 00002-11 LCB, Z01 DC 00035-03 LMG, 1 Z01 DC00056-01 LMG] NR 45 TC 25 Z9 27 U1 0 U2 0 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 2000 VL 10 IS 4 BP 487 EP 493 DI 10.1016/S0959-4388(00)00120-3 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 343JY UT WOS:000088700300009 PM 10981618 ER PT J AU Nguyen, MTT Arnheiter, H AF Nguyen, MTT Arnheiter, H TI Signaling and transcriptional regulation in early mammalian eye development: a link between FGF and MITF SO DEVELOPMENT LA English DT Article DE retinal pigment epithelium; neuroretina; neuroepithelium; optic vesicle culture; transcription factor; transcriptional repression; mouse; cell-fate specification ID RETINAL-PIGMENT EPITHELIUM; HELIX-ZIPPER TRANSCRIPTION; NEURAL RETINA; HOMEOBOX GENE; MELANOCYTE DEVELOPMENT; CELL-DIFFERENTIATION; LENS INDUCTION; NERVOUS-SYSTEM; MOUSE EMBRYO; CYCLIN D1 AB During vertebrate eye development, the optic vesicle is partitioned into a domain at its distal tip that will give rise to the neuroretina, and another at its proximal base that will give rise to the pigmented epithelium. Both domains are initially bipotential, each capable of giving rise to either neuroretina or pigmented epithelium, The partitioning depends on extrinsic signals, notably fibroblast growth factors, which emanate from the overlying surface ectoderm and induce the adjacent neuroepithelium to assume the neuroretinal fate, Using explant cultures of mouse optic vesicles, we demonstrate that bipotentiality of the optic neuroepithelium is associated with the initial coexpression of the basic-helix-loop-helix-zipper transcription factor MITF, which is later needed solely in the pigmented epithelium, and a set of distinct transcription factors that become restricted to the neuroretina. Implantation of fibroblast growth factor-coated beads close to the base of the optic vesicle leads to a rapid downregulation of MITF and the development of an epithelium that, by morphology, gene expression, and lack of pigmentation, resembles the future neuroretina. Conversely, the removal of the surface ectoderm results in the maintenance of MITF in the distal optic epithelium, lack of expression of the neuroretinal-specific CHX10 transcription factor, and conversion of this epithelium into a pigmented monolayer, This phenomenon can be prevented by the application of fibroblast growth factor alone. In Mitf mutant embryos, parts of the future pigment epithelium become thickened, lose expression of a number of pigment epithelium transcription factors, gain expression of neuroretinal transcription factors, and eventually transdifferentiate into a laminated second retina. The results support the view that the bipotential optic neuroepithelium is characterized by overlapping gene expression patterns and that selective gene repression, brought about by local extrinsic signals, leads to the separation into discrete expression domains and, hence, to domain specification. C1 Natl Inst Neurol Disorders & Stroke, Lab Dev Neurogenet, NIH, Bethesda, MD 20892 USA. RP Arnheiter, H (reprint author), Natl Inst Neurol Disorders & Stroke, Lab Dev Neurogenet, NIH, Bethesda, MD 20892 USA. NR 55 TC 173 Z9 175 U1 1 U2 6 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 2000 VL 127 IS 16 BP 3581 EP 3591 PG 11 WC Developmental Biology SC Developmental Biology GA 351NM UT WOS:000089165100016 PM 10903182 ER PT J AU Ishimura, A Maeda, R Takeda, M Kikkawa, M Daar, IO Maeno, M AF Ishimura, A Maeda, R Takeda, M Kikkawa, M Daar, IO Maeno, M TI Involvement of BMP-4/msx-1 and FGF pathways in neural induction in the Xenopus embryo SO DEVELOPMENT GROWTH & DIFFERENTIATION LA English DT Article DE bone morphogenetic protein; fibroblast growth factor; msx; neural induction; two-signal model ID BONE MORPHOGENETIC PROTEIN-4; FIBROBLAST GROWTH-FACTOR; SPEMANN ORGANIZER; NERVOUS-SYSTEM; CHICK-EMBRYO; EXPRESSION; INHIBITION; RECEPTOR; SIGNALS; NOGGIN AB The msx homeodomain protein is a downstream transcription factor of the bone morphogenetic protein (BMP)-4 signal and a key regulator for neural tissue differentiation. Xmsx-1 antagonizes the dorsal expression of noggin and cerberus, as revealed by in situ hybridization and reverse transcription-polymerase chain reaction assays. In animal cap explants, Xmsx-1 and BMP-4 inhibit the neural tissue differentiation induced by noggin or cerberus. A loss-of-function study using the Xmsx-1/VP-16 fusion construct indicated that neural tissue formation was directly induced by the injection of fusion ribonucleic acid, although the expression of neural cell adhesion molecule (N-CAM) in the cap was less than that in the cap injected with tBR or noggin. In contrast to the single cap assay, unexpectedly, both BMP-4 and Xmsx-1 failed to inhibit neurulation in the ectodermal explants to which the organizer mesoderm was attached. The results of cell-lineage tracing experiments indicated that the neural cells were differentiated from the animal pole tissue where the excess RNA of either BMP-4 or Xmsx-1 was injected, whereas notochord was differentiated from the organizer mesoderm. Neural tissue differentiated from BMP-4-injected ectodermal cells strongly expressed posterior neural markers, such as hoxB9 and krox20, suggesting that the posterior neural cells differentiated regardless of the existence of the BMP signal. The introduction of a dominant-negative form of the fibroblast growth factor (FGF) receptor (XFD) into the ectodermal cells drastically reduced the expression of pan and posterior neural markers (N-CAM and hoxB-9) if co-injected with BMP-4 RNA, although XFD alone at the same dose did not shut down the expression of N-CAM in the combination explants. Therefore, it is proposed that an FGF-related molecule was involved in the direct induction of posterior neural tissue in the inducing signals from the organizer mesoderm in vivo. C1 Niigata Univ, Fac Sci, Dept Biol, Niigata 9502181, Japan. NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Frederick, MD 21702 USA. RP Maeno, M (reprint author), Niigata Univ, Fac Sci, Dept Biol, 8050 Ikarashi-2, Niigata 9502181, Japan. OI Daar, Ira/0000-0003-2657-526X NR 51 TC 17 Z9 17 U1 0 U2 0 PU BLACKWELL SCIENCE ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 0012-1592 J9 DEV GROWTH DIFFER JI Dev. Growth Diff. PD AUG PY 2000 VL 42 IS 4 BP 307 EP 316 PG 10 WC Cell Biology; Developmental Biology SC Cell Biology; Developmental Biology GA 336LU UT WOS:000088304200002 PM 10969730 ER PT J AU Beanan, MJ Sargent, TD AF Beanan, MJ Sargent, TD TI Regulation and function of Dlx3 in vertebrate development SO DEVELOPMENTAL DYNAMICS LA English DT Review DE epidermis; placodes; distal-less genes; ectoderm development; homeodomain gene ID DISTAL-LESS GENE; HOMEOBOX GENES; EPIDERMAL DIFFERENTIATION; DEVELOPING FOREBRAIN; CORTICAL ROTATION; HOMEODOMAIN GENE; XENOPUS-LAEVIS; NEURAL CREST; 2 MEMBERS; EXPRESSION AB Dlx3 is a homeodomain transcription factor in vertebrates, related to Distal-less in Drosophila, that is expressed in differentiating epidermal cells, in neural crest, hair follicles, dental epithelium and mesenchyme, the otic and olfactory placodes, limb bud, placenta, and in the cement gland, which is located in the extreme anterior neural plate in Xenopus embryos. This factor behaves as a transcriptional activator, and positively regulates gene expression in the skin, and negatively regulates central nervous system markers in Xenopus epidermis and anterior neural plate. A mutation in the DLX3 gene is associated with a hereditary syndrome in humans, and loss of Dlx3 function is a developmental lethal in gene-targeted mice, where it is essential for proper modeling of the labyrinthine layer of the placenta. In this review, we discuss the evolution, expression, regulation, and function of Dlx3 in mouse, amphibians, and zebrafish, Published 2000 Wiley-Liss, Inc.dagger. C1 NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. RP Sargent, TD (reprint author), NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. NR 61 TC 57 Z9 60 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 1058-8388 J9 DEV DYNAM JI Dev. Dyn. PD AUG PY 2000 VL 218 IS 4 BP 545 EP 553 DI 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1026>3.0.CO;2-B PG 9 WC Anatomy & Morphology; Developmental Biology SC Anatomy & Morphology; Developmental Biology GA 339HA UT WOS:000088470500001 PM 10906774 ER PT J AU Xu, RH Peng, Y Fan, J Yan, DH Yamagoe, S Princler, G Sredni, D Ozato, K Kung, HF AF Xu, RH Peng, Y Fan, J Yan, DH Yamagoe, S Princler, G Sredni, D Ozato, K Kung, HF TI Histone acetylation is a checkpoint in FGF-stimulated mesoderm induction SO DEVELOPMENTAL DYNAMICS LA English DT Article DE histone; acetylation; FGF; trichostatin A; activator protein 1; mesoderm; Xenopus; embryo ID NF-KAPPA-B; EARLY XENOPUS DEVELOPMENT; FIBROBLAST GROWTH-FACTOR; TRANSCRIPTIONAL REPRESSION; EMBRYONIC-DEVELOPMENT; MAP KINASE; DEACETYLASE; EXPRESSION; HOMOLOG; INVOLVEMENT AB We have previously demonstrated that the transcription factor, AP-1 (c-jun/c-fos heterodimer), mediates fibroblast growth factor (FGF) signaling during mesoderm induction in Xenopus embryo. In the present studies, we show that histone acetylation is involved in FGF-mediated signaling leading to mesoderm induction, Histone acetylation is a dynamic process regulated by the activities of two histone-modifying enzymes, the histone acetyltransferase(s) and histone deacetylase(s) (HDACs), We found that basal and FGF-regulated activator protein 1 (AP-1) activity in Xenopus embryo is markedly reduced by treatment of trichostatin A (TSA), a specific inhibitor of HDAC. However, activity of another transcription factor, NF kappa B, is enhanced by TSA treatment. AP-1-mediated mesoderm induction in the animal caps is dramatically suppressed by TSA at a dose-dependent manner. This suppression can be rescued by ectopic expression of HDAC3 at early stage, Finally, are found that histone acetylation in animal caps is inhibited by FGF whereas enhanced by TSA las It control). Therefore, we propose that histone acetylation is a checkpoint for transduction of the FGF/AP-1 signals to induce mesoderm, Published 2000 Wiley-Liss, Inc.dagger. C1 Univ Wisconsin, Wisconsin Alumni Res Fdn, WiCell Res Inst, Madison, WI 53705 USA. NCI, Basic Res Lab, Div Basic Sci, Frederick Canc Res & Dev Ctr,NIH, Frederick, MD 21701 USA. NICHHD, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. Bar Ilan Univ, Canc Aids & Immunol Res Inst, Interdisciplinary Dept, Ramat Gan, Israel. NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. Univ Hong Kong, Inst Mol Biol, Pokfulam, Hong Kong, Peoples R China. RP Xu, RH (reprint author), Univ Wisconsin, Wisconsin Alumni Res Fdn, WiCell Res Inst, 614 Walnut St, Madison, WI 53705 USA. NR 44 TC 9 Z9 10 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 1058-8388 J9 DEV DYNAM JI Dev. Dyn. PD AUG PY 2000 VL 218 IS 4 BP 628 EP 635 DI 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1024>3.0.CO;2-H PG 8 WC Anatomy & Morphology; Developmental Biology SC Anatomy & Morphology; Developmental Biology GA 339HA UT WOS:000088470500008 PM 10906781 ER PT J AU Gabir, MM Hanson, RL Dabelea, D Imperatore, G Roumain, J Bennett, PH Knowler, WC AF Gabir, MM Hanson, RL Dabelea, D Imperatore, G Roumain, J Bennett, PH Knowler, WC TI The 1997 American Diabetes Association and 1999 World Health Organization criteria for hyperglycemia in the diagnosis and prediction of diabetes SO DIABETES CARE LA English DT Article ID OLDER ADULTS; GLUCOSE; MELLITUS AB OBJECTIVE - The 1997 American Diabetes Association (ADA) and the 1985 and 1999 World Health Organization (WHO) criteria for diabetes and hyperglycemia differ. The appropriateness of these diagnostic criteria in terms of individuals identified as abnormal and their prognosis has been debated. The purpose of this study is to compare the classifications of people by these criteria and to compare fasting and postload plasma glucose concentrations in the prediction of diabetes. RESEARCH DESIGN AND METHODS - The frequencies of diabetes by the 3 sets of criteria were compared in 5,023 adult Pima Indians not taking hypoglycemic drugs. Among nondiabetic subjects, Easting plasma glucose (FPG) and 2-h postload plasma glucose (2-h PG) concentrations and categories of impaired glucose regulation or diabetes were evaluated as predictors of diabetes defined by 1999 WHO criteria. RESULTS - The frequency of diabetes was 12.5% by 1997 ADA criteria, 14.6% by 1985 WHO criteria, and 15.3% by 1999 WHO criteria. The incidence of diabetes was strongly related to higher FPG and 2-h PG, each of which had very similar predictive powers. Impaired glucose tolerance (IGT) was more common than impaired fasting glucose (IFG) (15 vs. 5%), but the 5-year incidence of diabetes was higher in IFG than IGT (37 vs. 24%). CONCLUSIONS - The prevalence and incidence of diabetes are somewhat lower with the ADA criteria than with the 1985 or 1999 WHO criteria. The intermediate categories of glycemia differ substantially. IFG defines a smaller number of people who are at higher risk of developing diabetes than those with IGT. More people at high risk of diabetes could be identified by using either IFG or IGT, as recommended by the 1999 WHO criteria, or by using the FPG concentration alone, but with a lower cutoff value. C1 NIDDKD, NIH, Phoenix, AZ 85014 USA. RP Knowler, WC (reprint author), NIDDKD, NIH, 1550 E Indian Sch Rd, Phoenix, AZ 85014 USA. RI Hanson, Robert/O-3238-2015 OI Hanson, Robert/0000-0002-4252-7068 NR 15 TC 297 Z9 332 U1 3 U2 7 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD AUG PY 2000 VL 23 IS 8 BP 1108 EP 1112 DI 10.2337/diacare.23.8.1108 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 337RW UT WOS:000088376300010 PM 10937506 ER PT J AU Gabir, MM Hanson, RL Dabelea, D Imperatore, G Roumain, J Bennett, PH Knowler, WC AF Gabir, MM Hanson, RL Dabelea, D Imperatore, G Roumain, J Bennett, PH Knowler, WC TI Plasma glucose and prediction of microvascular disease and mortality - Evaluation of 1997 American Diabetes Association and 1999 World Health Organization criteria for diagnosis of diabetes SO DIABETES CARE LA English DT Article ID IMPAIRED FASTING GLUCOSE; PIMA-INDIANS; RISK FACTOR; CARDIOVASCULAR-DISEASE; BLOOD-GLUCOSE; HEART-DISEASE; OLDER ADULTS; TOLERANCE; MELLITUS; DEATH AB OBJECTIVE - The 1997 American Diabetes Association (ADA) and 1999 World Health Organization (WHO) criteria for diabetes and hyperglycemia were evaluated and compared with respect to prediction of microvascular and macrovascular disease and mortality. RESEARCH DESIGN AND METHODS - The prevalence of retinopathy and nephropathy at baseline and during the subsequent 10 years and mortality rates were examined in relation to baseline fasting plasma glucose (FPG) and 2-h postload plasma glucose (2-h PG) among 5,023 Pima Indian adults and in relation to the cut points defined by the ADA and WHO criteria. RESULTS - The frequencies of retinopathy and nephropathy were directly related to baseline FPG and 2-h PG with approximate thresholds near or below the current diagnostic criteria for diabetes (FPG greater than or equal to 7.0 and 2-h PG greater than or equal to 11.1 mmol/l). The rates of retinopathy were 4.7% in impaired fasting glucose (IFG) and 20.9% in diabetes by ADA criteria; 1.6% for impaired glucose tolerance (IGT) and 19.7% for diabetes by 1985 WHO criteria; and 1.2% for IGT and 19.2% for diabetes by the 1999 WHO criteria. Mortality rates from cardiovascular-renal-related diseases were higher in diabetic individuals (FPG greater than or equal to 7.0 or 2-h PG greater than or equal to 11.1 mmol/l) than in those with normal FPG and 2-h PG but were not elevated in those with IFG or IGT. CONCLUSIONS - Retinopathy and nephropathy were directly related to higher FPG or 2-h PG. FPG, which identifies those at high risk of microvascular disease and mortality, can be used to predict these outcomes and to diagnose diabetes when oral glucose tolerance testing is not practical. C1 NIDDKD, NIH, Phoenix, AZ 85014 USA. RP Knowler, WC (reprint author), NIDDKD, NIH, 1550 E Indian Sch Rd, Phoenix, AZ 85014 USA. RI Hanson, Robert/O-3238-2015 OI Hanson, Robert/0000-0002-4252-7068 NR 30 TC 102 Z9 117 U1 1 U2 4 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD AUG PY 2000 VL 23 IS 8 BP 1113 EP 1118 DI 10.2337/diacare.23.8.1113 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 337RW UT WOS:000088376300011 PM 10937507 ER PT J AU Brun, E Nelson, RG Bennett, PH Imperatore, G Zoppini, G Verlato, G Muggeo, M AF Brun, E Nelson, RG Bennett, PH Imperatore, G Zoppini, G Verlato, G Muggeo, M TI Diabetes duration and cause-specific mortality in the Verona Diabetes Study SO DIABETES CARE LA English DT Article ID CORONARY HEART-DISEASE; PIMA-INDIANS; RISK-FACTORS; POPULATION; MELLITUS; PREVALENCE; COHORT; DEATH; NIDDM AB OBJECTIVE - To examine the 10-year mortality and effect of diabetes duration on overall and cause-specific mortality in diabetic subjects in the Verona Diabetes Study (VDS). RESEARCH DESIGN AND METHODS - Records from diabetes clinics, family physicians, and a drug consumption database were used to identify 5,818 subjects greater than or equal to 45 years of age with type Z diabetes who were alive and residing in Verona, Italy on 31 December 1986. Vital status of each subject was ascertained on 31 December 1996. Underlying causes of death were determined from death certificates. Death rates and death rate ratios (DRRs) were computed and standardized to the population of Verona in 1991. RESULTS- During the study, 2,328 subjects died; 974 deaths were attributable to cardiovascular disease, 517 to neoplasms, 324 to diabetes-related diseases, 134 to digestive diseases, 250 to other natural causes, and 48 to external causes. There were 81 subjects who died of unknown causes. Death rates from natural causes were higher in men than in women (DRR 1.4, 95% CI 1.2-1.5) and rose in both sexes with increasing duration of diabetes (P = 0.001). Among the natural causes of death, those for diabetes-related diseases were strongly related to diabetes duration (P = 0.001); a modest relationship with duration was also found for ischemic heart disease in men (P = 0.07). CONCLUSIONS - Cardiovascular disease was the principal cause of death among people with type 2 diabetes in the VDS. Rates for natural causes of death rose with increasing duration of diabetes. Deaths from diabetes-related diseases in both sexes and from ischemic heart disease in men were largely responsible for this increase. C1 Univ Verona, Div Endocrinol & Metab Dis, I-37100 Verona, Italy. Univ Verona, Div Med Stat, I-37100 Verona, Italy. NIDDKD, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ USA. RP Muggeo, M (reprint author), Osped Civile Maggiore, Div Endocrinol & Malattie Metab, Piazzale Stefani 1, I-37126 Verona, Italy. RI SESM, SESM/C-1440-2008; Nelson, Robert/B-1470-2012 NR 24 TC 66 Z9 71 U1 2 U2 4 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD AUG PY 2000 VL 23 IS 8 BP 1119 EP 1123 DI 10.2337/diacare.23.8.1119 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 337RW UT WOS:000088376300012 PM 10937508 ER PT J AU Aspelund, G Egan, JM Slezak, LA Sritharan, KC Elahi, D Andersen, DK AF Aspelund, G Egan, JM Slezak, LA Sritharan, KC Elahi, D Andersen, DK TI Glucagon-like peptide-1 and exendin-4 improve glucose tolerance and induce islet cell growth in diabetic rats SO DIABETOLOGIA LA English DT Meeting Abstract C1 Yale Univ, Sch Med, Dept Surg, New Haven, CT 06510 USA. NIA, Gerontol Res Ctr, NIH, Diabet Sect, Baltimore, MD 21224 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Med, Boston, MA 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 2000 VL 43 SU 1 MA 555 BP A144 EP A144 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700553 ER PT J AU Egan, JM Clocquet, AR Greig, NH Hattersley, AT Habener, JF Tack, CJ Elahi, D AF Egan, JM Clocquet, AR Greig, NH Hattersley, AT Habener, JF Tack, CJ Elahi, D TI Decreased insulin secretion in response to glucose and GLP-1 in MODY-3 SO DIABETOLOGIA LA English DT Meeting Abstract C1 NIA, NIH, Baltimore, MD USA. Univ Exeter, Exeter EX4 4QJ, Devon, England. Catholic Univ Nijmegen, NL-6500 HC Nijmegen, Netherlands. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Boston, MA USA. RI Tack, Cees/A-2368-2014 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 2000 VL 43 SU 1 MA 67 BP A17 EP A17 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700067 ER PT J AU Klannemark, M Orho-Melander, M Rosberg, L Svensson, M Aberg, M Almgren, P Degerman, E Manganiello, V Groop, L AF Klannemark, M Orho-Melander, M Rosberg, L Svensson, M Aberg, M Almgren, P Degerman, E Manganiello, V Groop, L TI Discordant sibling-pair analysis of the phosphodiesterase 3B gene and association to postprandial hyperinsulinaemia SO DIABETOLOGIA LA English DT Meeting Abstract C1 Univ Lund, Dept Endocrinol, S-22100 Lund, Sweden. Univ Lund, Dept Cell & Mol Biol, S-22100 Lund, Sweden. NHLBI, Dept Pulm & Crit Care Med, NIH, Bethesda, MD 20892 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 2000 VL 43 SU 1 MA 204 BP A51 EP A51 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700205 ER PT J AU Michaeli, T Fletcher, C Jenkins, N Copeland, N Fleischer, N AF Michaeli, T Fletcher, C Jenkins, N Copeland, N Fleischer, N TI Glucose intolerance of mice with a targeted disruption of a cyclic amp specific phosphodiesterase. SO DIABETOLOGIA LA English DT Meeting Abstract C1 Albert Einstein Coll Med, Dept Dev & Mol Biol, Bronx, NY 10467 USA. Albert Einstein Coll Med, Dept Med, Bronx, NY 10467 USA. Albert Einstein Coll Med, Ctr Diabet Res & Training, Bronx, NY 10467 USA. NCI, Frederick Canc Res & Dev Ctr, Mammalian Genet Lab, Frederick, MD 21702 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 2000 VL 43 SU 1 MA 138 BP A35 EP A35 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700140 ER PT J AU Nelson, RG Lemley, KV Smith, WE Meyer, TW Bennett, PH Myers, BD AF Nelson, RG Lemley, KV Smith, WE Meyer, TW Bennett, PH Myers, BD TI Podocyte loss characterizes incipient nephropathy in Pima Indians with type 2 diabetes mellitus SO DIABETOLOGIA LA English DT Meeting Abstract C1 NIDDK, Good Samaritan Hosp, Phoenix, AZ USA. Stanford Univ, Stanford, CA 94305 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 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 2000 VL 43 SU 1 MA 103 BP A26 EP A26 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700103 ER PT J AU Notkins, AL Krause, M Odenwald, WF Toyama, R Cai, T AF Notkins, AL Krause, M Odenwald, WF Toyama, R Cai, T TI The IA-2 gene family: Homologs in C. elegans, Drosophila and zebrafish SO DIABETOLOGIA LA English DT Meeting Abstract C1 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 2000 VL 43 SU 1 MA 317 BP A80 EP A80 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700317 ER PT J AU Perrotti, N He, AR Lauro, D Taylor, SI AF Perrotti, N He, AR Lauro, D Taylor, SI TI Regulation of serum and glucocorticoid regulated protein kinase (Sgk) by cyclic amp and insulin. SO DIABETOLOGIA LA English DT Meeting Abstract C1 NIDDK, Diabet Branch, NIH, Bethesda, MD 20892 USA. Univ Catanzaro, Catanzaro, Italy. 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 2000 VL 43 SU 1 MA 181 BP A46 EP A46 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700182 ER PT J AU Tack, C Arioglu, E Lotsikas, A Papanicolaou, DA AF Tack, C Arioglu, E Lotsikas, A Papanicolaou, DA TI Circulating cytokines and insulin sensitivity; Effects of troglitazone SO DIABETOLOGIA LA English DT Meeting Abstract C1 Univ Med Ctr St Raboud, Div Gen Internal Med, Nijmegen, Netherlands. NIDDK, Diabet Branch, Bethesda, MD USA. NICHD, NIH, Dev Endocrinol Branch, Bethesda, MD USA. RI Tack, Cees/A-2368-2014 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 2000 VL 43 SU 1 MA 733 BP A191 EP A191 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700730 ER PT J AU Tataranni, PA Baier, L Jenkinson, C Harper, I Bogardus, C AF Tataranni, PA Baier, L Jenkinson, C Harper, I Bogardus, C TI A Ser311Cys substitution in the human dopamine receptor D2 is associated with reduced energy expenditure. SO DIABETOLOGIA LA English DT Meeting Abstract C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 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 2000 VL 43 SU 1 MA 125 BP A32 EP A32 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700126 ER PT J AU Weyer, C Tataranni, PA Bogardus, C Pratley, RE AF Weyer, C Tataranni, PA Bogardus, C Pratley, RE TI Insulin resistance and low early phase insulin secretion predict the progression from impaired glucose tolerance (IGT) to diabetes in Pima Indians. SO DIABETOLOGIA LA English DT Meeting Abstract C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 USA. NR 0 TC 8 Z9 8 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 2000 VL 43 SU 1 MA 285 BP A72 EP A72 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351BT UT WOS:000089136700285 ER PT J AU Glennon, RA Dukat, M Grella, B Hong, SS Costantino, L Teitler, M Smith, C Egan, C Davis, K Mattson, MV AF Glennon, RA Dukat, M Grella, B Hong, SS Costantino, L Teitler, M Smith, C Egan, C Davis, K Mattson, MV TI Binding of beta-carbolines and related agents at serotonin (5-HT2 and 5-HT1A), dopamine (D-2) and benzodiazepine receptors SO DRUG AND ALCOHOL DEPENDENCE LA English DT Article DE beta-carbolines; harmaline; harmine; hallucinogens; dimethyltryptamine (DMT); 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM); serotonin receptors; dopamine receptors; benzodiazepine receptors; PI hydrolysis ID ACID ETHYL-ESTER; DIFFERENTIAL INTERACTIONS; DISCRIMINATIVE STIMULUS; PARTIAL AGONIST; 1,2,3,4-TETRAHYDRO-BETA-CARBOLINES; IBOGAINE; ANALOGS; LSD AB A large series of beta-carbolines was examined for their ability to bind at [H-3]agonist-labeled 5-HT2A serotonin receptors. Selected beta-carbolines were also examined at 5-HT2C serotonin receptors, 5-MT1A serotonin receptors, dopamine D-2 receptors, and benzodiazepine receptors. Indolealkylamines and phenylisopropylamines were also evaluated in some of these binding assays. The beta-carbolines were found to bind with modest affinity at 5-HT2A receptors, and affinity was highly dependent upon the presence of ring substituents and ring saturation. The beta-carbolines displayed little to no affinity for 5-HT2A serotonin receptors, dopamine D-2 receptors and, with the exception of beta-CCM, for benzodiazepine receptors. Examples of beta-carbolines, indolealkylamines (i.e. N,N-dimethyltryptamine analogs), and phenylisopropylamines have been previously shown to produce common stimulus effects in animals trained to discriminate the phenylisopropylamine hallucinogen DOM (i.e. 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane) from vehicle. Although the only common receptor population that might account for this action is 5-HT2A, on the basis of a lack of enhanced affinity for agonist-labeled 5-HT2A receptors, as well as on their lack of agonist action in the PI hydrolysis assay, it is difficult to conclude that the beta-carbolines behave in a manner consistent with that of other classical hallucinogens. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved. C1 Virginia Commonwealth Univ, Sch Pharm, Dept Med Chem, Richmond, VA 23298 USA. Albany Med Coll, Dept Pharmacol, Albany, NY 12208 USA. NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Glennon, RA (reprint author), Virginia Commonwealth Univ, Sch Pharm, Dept Med Chem, Box 980540, Richmond, VA 23298 USA. RI Costantino, Luca/D-7608-2015 OI Costantino, Luca/0000-0001-5334-8084 FU NIDA NIH HHS [DA01642, DA09153] NR 35 TC 105 Z9 106 U1 1 U2 7 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0376-8716 J9 DRUG ALCOHOL DEPEN JI Drug Alcohol Depend. PD AUG 1 PY 2000 VL 60 IS 2 BP 121 EP 132 DI 10.1016/S0376-8716(99)00148-9 PG 12 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 337RY UT WOS:000088376500002 PM 10940539 ER PT J AU Weiss, RD Griffin, ML Gallop, R Onken, LS Gastfriend, DR Daley, D Crits-Christoph, P Bishop, S Barber, JP AF Weiss, RD Griffin, ML Gallop, R Onken, LS Gastfriend, DR Daley, D Crits-Christoph, P Bishop, S Barber, JP TI Self-help group attendance and participation among cocaine dependent patients SO DRUG AND ALCOHOL DEPENDENCE LA English DT Article DE self-help groups; Alcoholics Anonymous; cocaine; treatment; drug abuse ID NATIONAL-INSTITUTE; 12-STEP PROGRAMS AB The authors examined frequency and patterns of self-help group attendance and active participation over a 6-month period among 411 patients receiving treatment in the NIDA Collaborative Cocaine Treatment Study. Nearly two-thirds of patients attended at least one self-help group, and nearly all of these actively participated. Alcoholics Anonymous and Narcotics Anonymous meetings were attended most frequently. Statistical analyses included chi square, one-way analyses of variance, and cluster techniques. While patterns of attendance were relatively consistent over time, findings suggest that a treatment emphasizing the importance of self-help groups is likely to encourage more self-help group attendance and participation over time. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 McLean Hosp, Alcohol & Drug Abuse Treatment Program, Belmont, MA 02478 USA. Harvard Univ, Sch Med, Dept Psychiat, Boston, MA 02115 USA. Univ Penn, Sch Med, Philadelphia, PA 19104 USA. NIDA, Div Clin & Res Serv, Treatment Res Branch, Rockville, MD USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. Univ Pittsburgh, Western Psychiat Inst & Clin, Pittsburgh, PA 15213 USA. Brookside Hosp, Nashua, NH USA. RP Weiss, RD (reprint author), McLean Hosp, Alcohol & Drug Abuse Treatment Program, 115 Mill St, Belmont, MA 02478 USA. FU NIDA NIH HHS [K02DA00326, U01DA07090, U01DA07693, U18 DA007090] NR 21 TC 27 Z9 27 U1 2 U2 6 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0376-8716 J9 DRUG ALCOHOL DEPEN JI Drug Alcohol Depend. PD AUG 1 PY 2000 VL 60 IS 2 BP 169 EP 177 DI 10.1016/S0376-8716(99)00154-4 PG 9 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 337RY UT WOS:000088376500007 PM 10940544 ER PT J AU Echizen, H Tanizaki, M Tatsuno, J Chiba, K Berwick, T Tani, M Gonzalez, FJ Ishizaki, T AF Echizen, H Tanizaki, M Tatsuno, J Chiba, K Berwick, T Tani, M Gonzalez, FJ Ishizaki, T TI Identification of CYP3A4 as the enzyme involved in the mono-N-dealkylation of disopyramide enantiomers in humans SO DRUG METABOLISM AND DISPOSITION LA English DT Article ID HUMAN LIVER-MICROSOMES; REDUCTIVE METABOLISM; CATALYTIC PROPERTIES; DRUG-INTERACTIONS; IN-VITRO; MEPHENYTOIN; CYTOCHROME-P-450; HYDROXYLATION; EXPRESSION; IMIPRAMINE AB To identify which cytochrome P-450 (CYP) isoform(s) are involved in the major pathway of disopyramide (DP) enantiomers metabolism in humans, the in vitro formation of mono-N-desalkyldisopyramide from each DP enantiomer was studied with human liver microsomes and nine recombinant human CYPs. Substrate inhibition showed that SKF 525A and troleandomycin potently suppressed the metabolism of both DP enantiomers with IC50 values for R(-)- and S(+)-DP of <7.3 and <18.9 mu M, respectively. In contrast, only weak inhibitory effects (i.e., IC50 > 100 mu M) were observed for five other representative CYP isoform substrates [i.e., phenacetin (CYP1A1/2), sparteine (CYP2D6), tolbutamide (CYP2C9), S-mephenytoin (CYP2C19), and p-nitrophenol (CYP2E1)]. Significant correlations (P < .01, r = 0.91) were found between the activities of 11 different human liver microsomes for mono-N-dealkylation of both DP enantiomers and that of 6 beta-hydroxylation of testosterone. Conversely, no significant correlations were observed between the catalytic activities for DP enantiomers and those for the O-deethylation of phenacetin, 2-hydroxylation of desipramine, hydroxylation of tolbutamide, and 4'-hydroxylation of S-mephenytoin. Further evidence for involvement of CYP3A P450s was revealed by an anti-human CYP3A serum that inhibited the mono-N-dealkylation of both DP enantiomers and 6 beta-hydroxylation of testosterone almost completely (i.e., >90%), whereas it only weakly inhibited (i.e., <15%) CYP1A1/2- or 2C19-mediated reactions. Finally, the recombinant human CYP3A3 and 3A4 showed much greater catalytic activities than seven other isoforms examined (i.e., CYP1A2, 2A6, 2B6, 2C9, 2D6, 2E1, and 3A5) for both DP enantiomers. In conclusion, the metabolism of both DP enantiomers in humans would primarily be catalyzed by CYP3A4, implying that DP may have an interaction potential with other CYP3A substrates and/or inhibitors. C1 Meiji Pharmaceut Univ, Dept Pharmacotherapy, Tokyo 2048588, Japan. Sci Univ Tokyo, Dept Pharmaceut Sci, Tokyo 162, Japan. Mitsubishi Chem Corp, Tokyo, Japan. Int Med Ctr Japan, Div Gen Surg, Tokyo, Japan. Chiba Univ, Lab Biochem Pharmacol & Toxicol, Chiba, Japan. Kumamoto Univ, Grad Sch Clin Pharm, Dept Pharmacol & Therapeut, Kumamoto, Japan. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. RP Echizen, H (reprint author), Meiji Pharmaceut Univ, Dept Pharmacotherapy, Noshio 2-522-1, Tokyo 2048588, Japan. NR 40 TC 14 Z9 14 U1 0 U2 0 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 2000 VL 28 IS 8 BP 937 EP 944 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 335VY UT WOS:000088266500013 PM 10901704 ER PT J AU Lankford, SM Bai, SA Goldstein, JA AF Lankford, SM Bai, SA Goldstein, JA TI Cloning of canine cytochrome P450 2E1 CDNA: identification and characterization of two variant alleles SO DRUG METABOLISM AND DISPOSITION LA English DT Article ID ETHANOL-TREATED RABBITS; AMINO-ACID-SEQUENCES; LIVER CYTOCHROME-P-450; TRANSCRIPTIONAL REGULATION; GENETIC POLYMORPHISMS; MOLECULAR-CLONING; ESCHERICHIA-COLI; 2C SUBFAMILY; HUMAN CYP2E1; EXPRESSION AB Cytochrome P450 (CYP) 2E1 is a toxicologically important enzyme that inactivates a number of drugs and xenobiotics and also bioactivates many xenobiotic substrates to their hepatotoxic or carcinogenic forms. Although cDNAs for the human, rodent, and rabbit forms of CYP2E1 have been isolated and studied extensively, there is an absence of information about canine CYP2E1, despite the fact that the dog is routinely used in drug safety studies. In this study, we isolated and sequenced a full-length CYP2E1 cDNA from a beagle liver cDNA library. The deduced canine CYP2E1 amino acid sequence exhibited 75 to 76% identity with rat, mouse, and rabbit CYP2E1 sequences, and 77% identity with human CYP2E1. Two populations of clones, differing at a single nucleotide, were isolated from the unamplified library. The T1453C base change results in a Tyr(485)His amino acid substitution, which is well beyond the heme binding region but is possibly part of a beta-sheet structure. An allele-specific polymerase chain reaction-based restriction enzyme test was developed for genotyping individual dogs from genomic DNA samples. One hundred mixed breed dogs were genotyped, and the frequencies of the Tyr(485) and His(485) alleles were found to be 0.85 and 0.15, respectively. The canine Tyr(485) and His485 alleles and human CYP2E1 were expressed in Escherichia coli cells, and catalytic activities of the proteins were assessed using the substrate chlorzoxazone. Although the two canine enzymes had similar catalytic activity; significant kinetic differences were seen between canine and human CYP2E1s. C1 N Carolina State Univ, Coll Vet Med, Dept Anat Physiol Sci & Radiol, Raleigh, NC 27606 USA. DuPont Pharmaceut Co, Newark, DE USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Lankford, SM (reprint author), N Carolina State Univ, Coll Vet Med, Dept Anat Physiol Sci & Radiol, 4700 Hillsborough St, Raleigh, NC 27606 USA. RI Goldstein, Joyce/A-6681-2012 NR 38 TC 25 Z9 29 U1 0 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 2000 VL 28 IS 8 BP 981 EP 986 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 335VY UT WOS:000088266500019 PM 10901710 ER PT J AU Urnov, FD Yee, J Sachs, L Collingwood, TN Bauer, A Beug, H Shi, YB Wolffe, AP AF Urnov, FD Yee, J Sachs, L Collingwood, TN Bauer, A Beug, H Shi, YB Wolffe, AP TI Targeting of N-CoR and histone deacetylase 3 by the oncoprotein v-ErbA yields a chromatin infrastructure-dependent transcriptional repression pathway SO EMBO JOURNAL LA English DT Article; Proceedings Paper CT EMBO Workshop on Functional Organization of the Cell Nucleus CY AUG 09-12, 1999 CL PRAGUE, CZECH REPUBLIC SP EMBO DE histone deacetylase (HDAC)3; N-CoR; thyroid hormone receptor; transcriptional repression; v-ErbA ID THYROID-HORMONE RECEPTOR; TATA-BINDING PROTEIN; RETINOID-X-RECEPTOR; BETA-A GENE; NUCLEAR RECEPTOR; XENOPUS-LAEVIS; C-ERBA; FUNCTIONAL-ANALYSIS; ACTIVE REPRESSION; DNA METHYLATION AB Transcriptional repression by nuclear hormone receptors is thought to result from a unison of targeting chromatin modification and disabling the basal transcriptional machinery. We used Xenopus oocytes to compare silencing effected by the thyroid hormone receptor (TR) and its mutated version, the oncoprotein v-ErbA, on partly and fully chromatinized TR-responsive templates in vivo, Repression by v-ErbA was not as efficient as that mediated by TR, was significantly more sensitive to histone deacetylase (HDAC) inhibitor treatment and, unlike TR, v-ErbA required mature chromatin to effect repression. We find that both v-ErbA and TR can recruit the corepressor N-CoR, but, in contrast to existing models, show a concomitant enrichment for HDAC3 that occurs without an association with Sin3, HDAC1/ RPD3, Mi-2 or HDAC5. We propose a requirement for chromatin infrastructure in N-CoR/HDAC3-effected repression and suggest that the inability of v-ErbA to silence on partly chromatinized templates may stem from its impaired capacity to interfere with basal transcriptional machinery function. In support of this notion, we find v-ErbA to be less competent than TR for binding to TFIIB in vitro and in vivo. C1 Sangamo Biosci, Point Richmond Technol Ctr, Richmond, CA 94804 USA. NICHHD, Mol Embryol Lab, NIH, Bethesda, MD 20892 USA. Inst Mol Pathol, A-1030 Vienna, Austria. RP Wolffe, AP (reprint author), Sangamo Biosci, Point Richmond Technol Ctr, 501 Canal Blvd,Suite A100, Richmond, CA 94804 USA. NR 87 TC 60 Z9 61 U1 0 U2 1 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 2000 VL 19 IS 15 BP 4074 EP 4090 DI 10.1093/emboj/19.15.4074 PG 17 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 343BL UT WOS:000088681800023 PM 10921888 ER PT J AU Tujebajeva, RM Copeland, PR Xu, XM Carlson, BA Harney, JW Driscoll, DM Hatfield, DL Berry, MJ AF Tujebajeva, RM Copeland, PR Xu, XM Carlson, BA Harney, JW Driscoll, DM Hatfield, DL Berry, MJ TI Decoding apparatus for eukaryotic selenocysteine insertion SO EMBO REPORTS LA English DT Article ID ELONGATION-FACTOR SELB; TRANSFER-RNA; MAMMALIAN-CELLS; UGA; CODON; IDENTIFICATION; RECOGNITION; DEIODINASE; PROTEIN AB Decoding UGA as selenocysteine requires a unique tRNA, a specialized elongation factor, and specific secondary structures in the mRNA, termed SECIS elements. Eukaryotic SECIS elements are found in the 3' untranslated region of selenoprotein mRNAs while those in prokaryotes occur immediately downstream of UGA. Consequently, a single eukaryotic SECIS element can serve multiple UGA codons, whereas prokaryotic SECIS elements only function for the adjacent UGA, suggesting distinct mechanisms for recoding in the two kingdoms. We have identified and characterized the first eukaryotic selenocysteyl-tRNA-specific elongation factor. This factor forms a complex with mammalian SECIS binding protein 2, and these two components function together in selenocysteine incorporation in mammalian cells. Expression of the two functional domains of the bacterial elongation factor-SECIS binding protein as two separate proteins in eukaryotes suggests a mechanism for rapid exchange of charged for uncharged selenocysteyl-tRNA-elongation factor complex, allowing a single SECIS element to serve multiple UGA codons. C1 Brigham & Womens Hosp, Dept Med, Div Thyroid, Boston, MA 02115 USA. Harvard Univ, Sch Med, Boston, MA 02115 USA. Cleveland Clin Fdn, Lerner Res Inst, Dept Cell Biol, Cleveland, OH 44195 USA. NCI, Basic Res Lab, NIH, Bethesda, MD 20892 USA. RP Berry, MJ (reprint author), Harvard Inst Med, Div Thyroid, 77 Ave Louis Pasteur, Boston, MA 02115 USA. NR 19 TC 174 Z9 180 U1 2 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1469-221X J9 EMBO REP JI EMBO Rep. PD AUG PY 2000 VL 1 IS 2 BP 158 EP 163 DI 10.1093/embo-reports/kvd033 PG 6 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 381MD UT WOS:000165765900014 PM 11265756 ER PT J AU Hauache, OM Hu, JX Ray, K Spiegel, AM AF Hauache, OM Hu, JX Ray, K Spiegel, AM TI Functional interactions between the extracellular domain and the seven-transmembrane domain in Ca2+ receptor activation SO ENDOCRINE LA English DT Article DE calcium receptor; activation; functional; interations ID CALCIUM-SENSING RECEPTOR; CELL-SURFACE EXPRESSION; N-LINKED GLYCOSYLATION; SIGNAL-TRANSDUCTION; BINDING DOMAIN; DIMERIZATION; IDENTIFICATION; SITES AB We studied the activity of mutants involving the aminoterminal extracellular, seven-transmembrane (7TM) and carboxy-terminal tail domains of the human Ca2+ receptor to gain insight into the functional interactions between these domains during receptor activation. Missense mutations of highly conserved residues, D190 and E297, in the extracellular domain (ECD), and a mutation within part of the proximal carboxyterminal tail, A877-880E, resulted in receptors with severely reduced response to Ca2+ despite adequate cell surface expression. Coexpression of either D190A or E297K mutants with A877-880E led to significant reconstitution of function, No such reconstitution occurred when D190A or E297K mutants were coexpressed with a truncation mutant possessing an intact amino-terminal extracellular and first transmembrane domain, despite evidence for heterodimerization and cell surface expression of the respective mutant receptors, In addition, no reconstitution of function was observed when D190A was coexpressed with a deletion Ca2+ receptor mutant lacking only a cysteine-rich region located in the ECD of the Ca2+ receptor (Ca-//-Ca), Moreover, coexpression of this Ca-//-Ca with A877-880E did not recover function. The results show that Ca2+ receptor extracellular and 7TM domains are discrete entities that can communicate within the context of a heterodimer composed of complementary mutant receptors. Two intact 7TM domains and two intact cysteine-rich regions appear to be required for such communication to occur. The results are discussed in the context of a speculative model of receptor structure and function. C1 NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Spiegel, AM (reprint author), NIDDK, Metab Dis Branch, NIH, Bldg 31,Rm 9A-52, Bethesda, MD 20892 USA. NR 20 TC 11 Z9 12 U1 0 U2 1 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0969-711X J9 ENDOCRINE JI Endocrine PD AUG PY 2000 VL 13 IS 1 BP 63 EP 70 DI 10.1385/ENDO:13:1:63 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 351GC UT WOS:000089148600008 PM 11051048 ER PT J AU Bocchinfuso, WP Lindzey, JK Hewitt, SC Clark, JA Myers, PH Cooper, R Korach, KS AF Bocchinfuso, WP Lindzey, JK Hewitt, SC Clark, JA Myers, PH Cooper, R Korach, KS TI Induction of mammary gland development in estrogen receptor-alpha knockout mice SO ENDOCRINOLOGY LA English DT Article ID MOUSE MAMMARY; TARGETED DISRUPTION; PROLACTIN GENE; GROWTH-HORMONE; CYCLIN D1; WILD-TYPE; ER-ALPHA; PROGESTERONE; PITUITARY; BETA AB Mammary glands from the estrogen receptor-a! knockout (alpha ERKO) mouse do not undergo ductal morphogenesis or alveolar development. Disrupted ER alpha signaling may result in reduced estrogen-responsive gene products in the mammary gland or reduced mammotropic hormones that contribute to the alpha ERKO mammary phenotype. We report that circulating PRL is reduced in the female alpha ERKO mouse. Implantation of an age-matched, heterozygous ER alpha pituitary isograft under the renal capsule of 25-day-old or 12-week-old alpha ERKO mice increased circulating PRL and progesterone levels, and induced mammary gland development. Grafted alpha ERKO mice also possessed hypertrophied corpora lutea demonstrating that PRL is luteotropic in the alpha ERKO ovary. By contrast, ovariectomy at the time of pituitary grafting prevented mammary gland development in alpha ERKO mice despite elevated PRL levels. Hormone replacement using pellet implants demonstrated that pharmacological doses of estradiol induced limited mammary ductal elongation, and estradiol in combination with progesterone stimulated lobuloalveolar development. PRL alone or in combination with progesterone or estradiol did not induce alpha ERKO mammary growth. Estradiol and progesterone are required for the structural development of the alpha EBKO mammary gland, and PRL contributes to this development by inducing ovarian progesterone levels. Therefore, the manifestation of the alpha ERKO mammary phenotype appears due to the lack of direct estrogen action at the mammary gland and an indirect contributory role of estrogen signaling at the hypothalamic/pituitary axis. C1 NIEHS, Reprod & Dev Toxicol Lab, Receptor Biol Sect, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Comparat Med Branch, NIH, Res Triangle Pk, NC 27709 USA. US EPA, Reprod Toxicol Div, Res Triangle Pk, NC 27711 USA. Univ S Florida, Dept Biol, Tampa, FL 33620 USA. RP Korach, KS (reprint author), NIEHS, Reprod & Dev Toxicol Lab, Receptor Biol Sect, NIH, POB 12233,MD B302, Res Triangle Pk, NC 27709 USA. EM korach@niehs.nih.gov OI Korach, Kenneth/0000-0002-7765-418X NR 63 TC 134 Z9 139 U1 2 U2 5 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD AUG PY 2000 VL 141 IS 8 BP 2982 EP 2994 DI 10.1210/en.141.8.2982 PG 13 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 337WP UT WOS:000088386300034 PM 10919287 ER PT J AU Winterer, G Ziller, M Dorn, H Frick, K Mulert, C Wuebben, Y Herrmann, WM AF Winterer, G Ziller, M Dorn, H Frick, K Mulert, C Wuebben, Y Herrmann, WM TI Frontal dysfunction in schizophrenia - a new electrophysiological classifier for research and clinical applications SO EUROPEAN ARCHIVES OF PSYCHIATRY AND CLINICAL NEUROSCIENCE LA English DT Article DE schizophrenia; classification; EEG; event-related potentials; hypofrontality ID EEG; SCALE AB We determined whether schizophrenic patients can be reliably classified with electrophysiological tools. We developed a fully computerized classifier based on 5 minutes of EEG recording during an acoustical choice reaction time task (AMDP-module IV). We included factorized variables from the frequency domain and evoked potentials (N100/P200-complex) from central and frontal electrodes, which were preprocessed in a sample of 150 normal subjects prior to classification. We applied discriminant analyses to the electrophysiological data from depressive, schizophrenic and schizotypal subjects, most of them being unmedicated or drug-naive. The classifier was developed on a training set (33 schizophrenics, 49 normals) and tested on an independent sample (32 schizophrenics, 49 normals). A simple three-variable classifier was found to classify schizophrenics and normals in 77 % of those tested correctly. Diagnostic specificity of the das sifier proved to be low as the inclusion of depressive patients (n = 60) significantly decreased classification power. It was demonstrated that atypical but not typical neuroleptic drugs may influence the classification results. Correctly classified schizophrenics showed significantly more negative symptoms and slower reaction times than those schizophrenics who were misclassified as normals. In contrast, these misclassified schizophrenics showed a non-significant trend for more positive symptoms and shorter reaction times. As the correctly classified schizophrenics showed increased frontally pronounced delta-activity and decreased signal power of the N100/P200 amplitude, it was concluded that these schizophrenics show dysfunction of the frontal lobe. It is proposed that this new classifier can be useful for clinical and research applications when subtyping of schizophrenics with detection of frontal dysfunction as the aim. C1 NIMH, Clin Brain Disorder Branch, NIH, Bethesda, MD 20892 USA. Free Univ Berlin, Hosp Benjamin Franklin, Dept Psychiat, D-1000 Berlin, Germany. RP Winterer, G (reprint author), NIMH, Clin Brain Disorder Branch, NIH, Bldg 10,Room 4s229A MSC 1379, Bethesda, MD 20892 USA. RI Mulert, Christoph/F-2576-2012 NR 26 TC 37 Z9 39 U1 1 U2 1 PU DR DIETRICH STEINKOPFF VERLAG PI DARMSTADT PA PLATZ DER DEUTSCHEN EINHEIT 25, D-64293 DARMSTADT, GERMANY SN 0940-1334 J9 EUR ARCH PSY CLIN N JI Eur. Arch. Psych. Clin. Neurosci. PD AUG PY 2000 VL 250 IS 4 BP 207 EP 214 DI 10.1007/s004060070026 PG 8 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 350GQ UT WOS:000089093000007 PM 11009074 ER PT J AU Feucht, J Aigner, S Ehler, E Suter, TM Adelstein, RS Hess, OM Eppenberger, HM AF Feucht, J Aigner, S Ehler, E Suter, TM Adelstein, RS Hess, OM Eppenberger, HM TI Localization and expression levels of nonmuscle myosin heavy chain isoformes in heart tissue and in cultivated ventricular adult rat cardiomyocytes SO EUROPEAN HEART JOURNAL LA English DT Meeting Abstract C1 Swiss Fed Inst Technol, Inst Cell Biol, Zurich, Switzerland. NIH, Bethesda, MD 20892 USA. Univ Hosp Bern, Swiss Cardiovasc Ctr, CH-3010 Bern, Switzerland. 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 2000 VL 21 SU S MA P455 BP 57 EP 57 PG 1 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 351BR UT WOS:000089136600218 ER PT J AU Howcroft, TK AF Howcroft, TK TI HIV-1: a molecular toolkit for analysis of major histomcompatibility complex class I expression SO EUROPEAN JOURNAL OF CLINICAL INVESTIGATION LA English DT Editorial Material ID MHC CLASS-I; CYTOTOXIC T-LYMPHOCYTES; DOWN-REGULATION; NEF PROTEIN; CELL RECEPTORS; HLA-E; TAT; TRANSCRIPTION; SIGNAL; TRAFFICKING C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Howcroft, TK (reprint author), NCI, Expt Immunol Branch, NIH, Bldg 10,Room 4B-17,10 Ctr Dr MSC 1360, Bethesda, MD 20892 USA. NR 28 TC 2 Z9 2 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2972 J9 EUR J CLIN INVEST JI Eur. J. Clin. Invest. PD AUG PY 2000 VL 30 IS 8 BP 657 EP 659 PG 3 WC Medicine, General & Internal; Medicine, Research & Experimental SC General & Internal Medicine; Research & Experimental Medicine GA 347PA UT WOS:000088937300002 PM 10964156 ER PT J AU Chiodetti, L Barber, DL Schwartz, RH AF Chiodetti, L Barber, DL Schwartz, RH TI Biallelic expression of the IL-2 locus under optimal stimulation conditions SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE T lymphocyte; cytokine; limiting dilution; intracellular staining ID T-CELL DIFFERENTIATION; GENE-EXPRESSION; INTERLEUKIN-2 TRANSCRIPTION; ANTIGEN RECEPTOR; MESSENGER-RNA; MICE; HETEROGENEITY; ACTIVATION; ALLELES; PATHWAY AB Recent experiments have suggested that the IL-2 locus is monoallelically expressed. We tested this hypothesis using TCR-transgenic mice carrying one inactivated IL-2 allele. The frequency in single-cell assays of IL-2-producing cells following optimal stimulation by antigen and antigen-presenting cells was equivalent to that from wild-type mice, but the amount of IL-2 produced per cell was twofold less. Similar observations were made by intracellular staining for IL-2, although stimulation in bulk culture was less optimal, showing only a 1.7-fold difference. Importantly, the frequency of responding cells from the heterozygotes was less than from the wild-type mice if the IL-2 assay was performed after only 24-30 h of activation, suggesting that the targeted allele could compete with the normal allele early after stimulation and give the misimpression that the heterozygotes had fewer IL-2-producing cells. These data strongly argue that the IL-2 locus can be expressed biallelically under optimum stimulation conditions. C1 NIAID, Cellular & Mol Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Chiodetti, L (reprint author), NIAID, Cellular & Mol Immunol Lab, NIH, Bldg 4,Rm 111, Bethesda, MD 20892 USA. NR 26 TC 14 Z9 14 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI BERLIN PA MUHLENSTRASSE 33-34, D-13187 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD AUG PY 2000 VL 30 IS 8 BP 2157 EP 2163 DI 10.1002/1521-4141(2000)30:18<2157::AID-IMMU2157>3.0.CO;2-G PG 7 WC Immunology SC Immunology GA 346DZ UT WOS:000088856700006 PM 10940906 ER PT J AU Papadopoulos, EJ Fitzhugh, DJ Tkaczyk, C Gilfillan, AM Sassetti, C Metcalfe, DD Hwang, ST AF Papadopoulos, EJ Fitzhugh, DJ Tkaczyk, C Gilfillan, AM Sassetti, C Metcalfe, DD Hwang, ST TI Mast cells migrate, but do not degranulate, in response to fractalkine, a membrane-bound chemokine expressed constitutively in diverse cells of the skin SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE chemokine; mast cell; skin; trafficking ID DENDRITIC CELLS; SENSORY NERVES; CX3C CHEMOKINE; RECEPTOR; CC; HEXOSAMINIDASE; ADHESION; RELEASE AB Mast cells (MC) are anatomically located near nerves acid blood vessels in skin and the gastrointestinal tract and tend to localize within certain cutaneous tumors such as neurofibromas, However, the molecular mechanisms by which MC home to these sites are not well characterized. Fractalkine (FK) is a membrane-bound CX3C chemokine that displays constitutive expression in dendritic cells as well as in non-hematopoietic tissues including mammalian brain. Here we show that FK is constitutively expressed by skin endothelial cells, dermal dendrocytes and cells within neurofibromas. By reverse transcription-PCR, FK receptor, CX3CR1, is expressed by cultured murine bone marrow-derived MC (BMMC) of both connective tissue and mucosal phenotypes. Non-activated human dermal MC isolated from neonatal foreskin similarly demonstrated CX3CR1 expression. In chemotaxis assays, FK attracted MC with maximal migration occurring between 25-125 ng/ml. BMMC were not stimulated to release proinflammatory mediators in the presence of FK as measured by granule-associated beta-hexosaminidase release, Thus, CX3CR1 is expressed by MC and effectively mediates chemotaxis without inducing degranulation. We propose that the constitutive expression of FK on certain cells in the skin may be a factor in the tissue-specific homing of MC. C1 NCI, Dermatol Branch, Bethesda, MD 20892 USA. NIAID, Lab Allerg Dis, Bethesda, MD 20892 USA. Univ Calif San Francisco, Dept Anat, San Francisco, CA 94143 USA. RP Hwang, ST (reprint author), NCI, Dermatol Branch, Bldg 10,Rm 12N246,10 Ctr Dr,MSC 1908, Bethesda, MD 20892 USA. NR 31 TC 62 Z9 63 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI BERLIN PA MUHLENSTRASSE 33-34, D-13187 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD AUG PY 2000 VL 30 IS 8 BP 2355 EP 2361 DI 10.1002/1521-4141(2000)30:8<2355::AID-IMMU2355>3.0.CO;2-# PG 7 WC Immunology SC Immunology GA 346DZ UT WOS:000088856700026 PM 10940926 ER PT J AU Andringa, G van Oosten, RV Unger, W Hafmans, TGM Veening, J Stoof, JC Cools, AR AF Andringa, G van Oosten, RV Unger, W Hafmans, TGM Veening, J Stoof, JC Cools, AR TI Systemic administration of the propargylamine CGP 3466B prevents behavioural and morphological deficits in rats with 6-hydroxydopamine-induced lesions in the substantia nigra SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE 6-hydroxydopamine; apoptosis; behaviour; neuroprotection; Parkinson's disease ID MESENCEPHALIC DOPAMINE NEURONS; PARKINSONS-DISEASE; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; TYROSINE-HYDROXYLASE; SPATIAL NAVIGATION; DYING NEURONS; ANIMAL-MODEL; L-DEPRENYL; IN-VITRO; SELEGILINE AB The ability of CGP 3466B to attenuate the behavioural and morphological consequences of experimentally induced cell death was investigated in a recently updated animal model of Parkinson's disease. 6-Hydroxydopamine was infused bilaterally into the substantia nigra pars compacta of rats that were pretreated with desimipramine. Treatment with CGP 3466B (0.0014-1.4 mg/kg, injected subcutaneously) or its solvent was begun 2 h after the 6-OHDA injection, and maintained twice daily for 14 days. After a washout period of 14 days, changes in motor behaviour were evaluated, using the open field test (analysis of normal and abnormal stepping, e.g.) and the paw test (analysis of retraction time of limbs). Changes in learning and memory were evaluated with the help of the Morris water maze task. Following immunocytochemical staining of tyrosine hydroxylase, the extent of the lesion was quantified using a computerized system. CGP 3466B prevented all deficits produced by 6-hydroxydopamine (6-OHDA), though at different doses. It prevented: abnormal stepping (0.0014-0.014 mg/kg); increased forelimb and hindlimb retraction time (0.014-0.14 mg/kg and 0.0014-0.14 mg/kg, respectively); delayed learning (1.4 mg/kg); and reduced tyrosine hydroxylase immunoreactivity in the substantia nigra (0.0014-0.014 mg/kg). CGP 3466B (0.0014-0.14 mg/kg) induced no deficits in sham-treated rats. CGP 3466B (1.4 mg/kg), however, did not show any benefit on motor deficits in 6-OHDA-lesioned rats, and induced abnormal movements and decreased the tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and the ventral tegmental area of sham-lesioned animals. It is concluded that CGP 3466B prevents all 6-OHDA-induced behavioural and immunocytochemical deficits, though at different doses. CGP 3466B is suggested to be a valuable agent for inhibiting the dopaminergic degeneration in patients with Parkinson's disease. C1 Univ Nijmegen, Dept Psychoneuropharmacol, Nijmegen, Netherlands. Univ Nijmegen, Dept Anat & Embryol, Nijmegen, Netherlands. Vrije Univ Amsterdam, Dept Neurol, Neurosci Res Inst, Amsterdam, Netherlands. RP Andringa, G (reprint author), NINDS, Expt Therapeut Branch, NIH, Bldg 10,Rm 5C215,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Unger, Wendy/G-8767-2011 NR 64 TC 35 Z9 38 U1 0 U2 3 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD AUG PY 2000 VL 12 IS 8 BP 3033 EP 3043 DI 10.1046/j.1460-9568.2000.00181.x PG 11 WC Neurosciences SC Neurosciences & Neurology GA 349PP UT WOS:000089055100036 PM 10971644 ER PT J AU MacDougall, M Simmons, D Gu, TT Forsman-Semb, K Mardh, CK Mesbah, M Forest, N Krebsbach, PH Yamada, Y Berdal, A AF MacDougall, M Simmons, D Gu, TT Forsman-Semb, K Mardh, CK Mesbah, M Forest, N Krebsbach, PH Yamada, Y Berdal, A TI Cloning, characterization and immunolocalization of human ameloblastin SO EUROPEAN JOURNAL OF ORAL SCIENCES LA English DT Article DE ameloblastin; amelogenesis imperfecta; ameloblasts; alternative splicing; sheathlin; amelin ID DOMINANT AMELOGENESIS IMPERFECTA; AUTOSOMAL-DOMINANT; SHEATH PROTEINS; CHROMOSOME 4Q; RAT INCISORS; GENE; ENAMEL; EXPRESSION; REGION; LOCALIZATION AB Amelogenesis imperfecta is a broad classification of hereditary enamel defects, exhibiting both genetic and clinical diversity. Most amelogenesis imperfecta cases are autosomal dominant disorders, yet only the local hypoplastic form has been mapped to human chromosome 4q between D4S2421 and the albumin gene. An enamel protein cDNA, termed ameloblastin (also known as amelin and sheathlin), has been isolated from rat, mouse and pig. Its human homolog has been mapped to chromosome 4q21 between markers D4S409 and D4S400, flanking the local hypoplastic amelogenesis imperfecta critical region. Therefore, ameloblastin is a strong candidate gene for this form of amelogenesis imperfecta. To facilitate genetic studies related to this dental disease, we isolated and characterized a human ameloblastin cDNA. A human third molar cDNA library was screened and two ameloblastin clones identified. Nucleotide sequencing of these cDNAs indicated alternative splicing of the putative open reading frame, use of different polyadenylation signals, and a high degree of similarity to reported rat, mouse and porcine cDNAs. Immunohistochemistry studies on embryonic human teeth using an antibody to recombinant ameloblastin indicated ameloblastin expression by ameloblasts with localization in the enamel matrix associated with the sheath structures. C1 Univ Texas, Hlth Sci Ctr, Sch Dent, San Antonio, TX 78229 USA. Umea Univ, Dept Clin Genet, S-90187 Umea, Sweden. Umea Univ, Dept Appl Cell & Mol Biol, S-90187 Umea, Sweden. Univ Michigan, Sch Dent, Ann Arbor, MI 48109 USA. Natl Inst Dent & Craniofacial Res, Bethesda, MD USA. Univ Paris 07, Cordeliers Inst, Fac Dent, Lab Biol Odontol, Paris, France. RP MacDougall, M (reprint author), Univ Texas, Hlth Sci Ctr, Sch Dent, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA. FU NIDCR NIH HHS [DE09875] NR 23 TC 30 Z9 41 U1 1 U2 4 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0909-8836 J9 EUR J ORAL SCI JI Eur. J. Oral Sci. PD AUG PY 2000 VL 108 IS 4 BP 303 EP 310 DI 10.1034/j.1600-0722.2000.108004303.x PG 8 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 339KV UT WOS:000088477000007 PM 10946765 ER PT J AU Gordis, E AF Gordis, E TI Contributions of behavioral science to alcohol research: Understanding who is at risk and why SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material ID MUTANT MICE LACKING; FOLLOW-UP; DEPENDENCE; NALTREXONE; ETHANOL; RECEPTORS; THERAPY; ABUSE; DRUG AB Behavioral science has been an active participant in alcohol research progress over the past 30 years, particularly in the areas of prevention and treatment methodology. However, alcoholism results from the interaction between complex biological and behavioral systems, and in recent years, combined behavioral and biological studies, primarily of alcohol effects on the brain and of the genetics of alcoholism, have begun the much more complex process of elucidating the links between biology and specific alcohol use behaviors. It is this combined research that ultimately will produce the pharmacological and behavioral interventions that will improve the efficiency and effectiveness of alcohol prevention and treatment methods. C1 NIAAA, NIH, Bethesda, MD USA. RP Gordis, E (reprint author), NIAAA, NIH, Bethesda, MD USA. NR 19 TC 8 Z9 8 U1 3 U2 3 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 2000 VL 8 IS 3 BP 264 EP 270 DI 10.1037//1064-1297.8.3.264 PG 7 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 348ZD UT WOS:000089015600002 PM 10975614 ER PT J AU Hyman, SE AF Hyman, SE TI National Institute of Mental Health goals for behavioral science SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material AB Because all health conditions for which the National Institute of Mental Health (NIMH) is responsible manifest at the level of behavior, and all interventions must have an impact at the behavioral level, NIMH is firmly committed to the support of behavioral science. In an era in which research in areas that some view as reductionist-for example, genomics, genetics, functional genomics and proteomics, and molecular science-is especially promising, NIMH is striving to maintain a balance in its portfolio with studies that explore integrative aspects of biology, including behavior. Without this perspective, new information about fundamental processes will prove ultimately to be shallow. This commentary discusses how understanding of brain and behavior in mental illness and health calls for integrating bottom-up research that studies brain and behavior through genes and molecules, with top-down research that examines the impact of environment. C1 NIMH, Bethesda, MD 20892 USA. RP Hyman, SE (reprint author), NIMH, 6001 Execut Bldg,Room 8235 MSC 9669, Bethesda, MD 20892 USA. NR 0 TC 3 Z9 3 U1 0 U2 0 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 2000 VL 8 IS 3 BP 271 EP 272 PG 2 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 348ZD UT WOS:000089015600003 PM 10975615 ER PT J AU Leshner, AI AF Leshner, AI TI National Institute on Drug Abuse's behavioral research agenda SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material AB Basic behavioral research continues to play an integral role in the National Institute on Drug Abuse's (NIDA's) search for solutions to the complex social and public health problems posed by drug abuse and addiction. Along with NIDA's basic molecular and neuroscience research programs, behavioral research has played an important role in increasing clinician's understanding of the mechanisms and processes that underlie addiction. Much has been learned about the ways in which animals and humans respond to their environment and the role these basic behavioral processes play in drug abuse and other drug-abuse-related phenomena, such as withdrawal, craving, and relapse, but there is still much more to be known. The author discusses how NIDA will continue to build and promote its behavioral research agenda and ensure that behavioral research findings are applied in real-life settings when applicable. C1 NIDA, Bethesda, MD 20892 USA. RP NIDA, 6001 Execut Blvd,Room 5230,MSC 9591, Bethesda, MD 20892 USA. EM leshner@nih.gov NR 0 TC 3 Z9 3 U1 0 U2 0 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 EI 1936-2293 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD AUG PY 2000 VL 8 IS 3 BP 273 EP 275 DI 10.1037//1064-1297.8.3.273 PG 3 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 348ZD UT WOS:000089015600004 PM 10975616 ER PT J AU Robinson, ML Houtsmuller, EJ Moolchan, ET Pickworth, WB AF Robinson, ML Houtsmuller, EJ Moolchan, ET Pickworth, WB TI Placebo cigarettes in smoking research SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Review ID DE-NICOTINIZED CIGARETTES; EEG CHANGES; TOBACCO WITHDRAWAL; SUBJECTIVE RATINGS; SMOKERS; PERFORMANCE; REPLACEMENT; ABSTINENCE; DELIVERY; BEHAVIOR AB This review outlines the development and use of placebo cigarettes in smoking research. Research on effects of smoking has been disadvantaged by the lack of an adequate placebo condition. Recently, tobacco-based denicotinized cigarettes have been used in smoking research to distinguish effects of smoking due to the delivery of nicotine, other components of tobacco smoke, and the sensory process of smoking. Placebo cigarettes do not increase heart rate and blood pressure or produce electroencephalogram changes ordinarily associated with nicotine. However, placebo cigarettes reduce subjective measures of tobacco craving, desire to smoke, and tobacco withdrawal. These findings indicate that the effects of cigarette smoking are dependent on the delivery of nicotine, tar, other compounds of tobacco smoke, and the sensory stimuli. The next generation of research may begin to investigate the mechanisms that modulate these placebo effects. C1 NIDA, IRP, Baltimore, MD 21224 USA. RP Pickworth, WB (reprint author), NIDA, IRP, 5500 Nathan Shock Dr,POB 5180, Baltimore, MD 21224 USA. NR 50 TC 37 Z9 37 U1 7 U2 10 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 2000 VL 8 IS 3 BP 326 EP 332 DI 10.1037//1064-1297.8.3.326 PG 7 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 348ZD UT WOS:000089015600008 PM 10975620 ER PT J AU Shurtleff, D AF Shurtleff, D TI A behavioral economic analysis of the relative persistence of behavior: Comment on Meisch (2000) SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material ID RHESUS-MONKEYS; DRUG; DEMAND; PRICE; COST; FOOD AB This commentary examines and reinterprets the concept of relative persistence in drug self-administration studies, described by R. A. Meisch (2000), in behavioral economic terms. Over the past several years, investigators in the behavioral sciences have successfully applied consumer demand theory to the study of drug abuse and addiction. The economic concept of demand elasticity (i.e., the changes in the amount of a commodity demanded as a function of changes in price) and the concept of unit price are described in detail, and this commentary shows these concepts provide an alternative interpretation to the relative persistence of behavior. The application of the behavioral economic approach to understanding abuse potential of putative drugs of abuse, in development of medications for drug addiction and in characterizing the transition from drug use to drug addiction, is discussed. C1 NIDA, Div Neurosci & Behav Res, Bethesda, MD 20892 USA. RP Shurtleff, D (reprint author), NIDA, Div Neurosci & Behav Res, 6001 Execut Blvd,Room 4282,MSC 9555, Bethesda, MD 20892 USA. NR 15 TC 2 Z9 2 U1 0 U2 0 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 2000 VL 8 IS 3 BP 357 EP 359 DI 10.1037//1064-1297.8.3.357 PG 3 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 348ZD UT WOS:000089015600013 PM 10975625 ER PT J AU Taraboletti, G Sonzogni, L Vergani, V Hosseini, G Ceruti, R Ghilardi, C Bastone, A Toschi, E Borsotti, P Scanziani, E Giavazzi, R Pepper, MS Stetler-Stevenson, WG Bani, MR AF Taraboletti, G Sonzogni, L Vergani, V Hosseini, G Ceruti, R Ghilardi, C Bastone, A Toschi, E Borsotti, P Scanziani, E Giavazzi, R Pepper, MS Stetler-Stevenson, WG Bani, MR TI Posttranscriptional stimulation of endothelial cell matrix metalloproteinases 2 and 1 by endothelioma cells SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE matrix metalloproteinases; endothelial cells; endothelioma; angiogenesis; hemangioma; proteases secretion ID MIDDLE-T-ONCOGENE; TISSUE INHIBITOR; INTERSTITIAL COLLAGENASE; PLASMINOGEN-ACTIVATOR; MOUSE DEVELOPMENT; IV COLLAGENASE; GELATINASE-B; ANGIOGENESIS; GROWTH; EXPRESSION AB Matrix metalloproteinases (MMPs) play a critical role in the development of hemangioma-like vascular tumors in mice injected with murine eEnd.1 endothelioma cells. The current study was designed to (a) characterize the presence of MMPs in the vascular tumor, (b) define whether these MMPs originate from the transformed cells or from the recruited stromal cells and (c) study the stimulatory effect of eEnd.1 cells on the production of MMPs by endothelial cells. Several gelatinases were present in the eEnd.1 tumor extract, including latent and activated MMP-2 (72-kDa gelatinase A, EC 3.4.24.24) and MMP-9 (92-kDa gelatinase B, EC 3.4.24.35). Immunohistochemical analysis of the tumor revealed focal reactivity for MMP-2. No gelatinase was produced by cultured eEnd.1 cells, or by six of nine related endothelioma cell lines, suggesting that stroma cells, particularly endothelial cells recruited by the tumor cells, rather than eEnd.1 cells themselves, are the source of the gelatinases observed in the tumors in vivo. The conditioned medium of eEnd.1 cells stimulated the release of MMP-2 and MMP-1 (interstitial collagenase, EC 3.4.24.7) by endothelial cells, but not of the inhibitor TIMP-2. The increased production of MMP-2 and MMP-1, observed at the protein level (zymogram and Western blot analysis), occurred through a posttranscriptional mechanism, since no increase in mRNA was observed and the stimulation was not prevented by inhibitors of protein synthesis. The inhibitory effects of monensin and brefeldin A, inhibitors of protein secretion, and the decrease in cell-associated MMP-2 in stimulated endothelial cells indicated that regulation occurred mostly at the level of protease secretion. MMPs are known to be regulated at different levels; this study indicates that, in endothelial cells, the stimulation of MMPs can also occur at the level of secretion, a mechanism that provides a rapid mobilization of these crucial enzymes in the early phases of angiogenesis. (C) 2000 Academic Press. C1 Mario Negri Inst Pharmacol Res, Dept Oncol, I-24125 Bergamo, Italy. Univ Milan, Ist Anat Patol Vet & Patol Aviare, Milan, Italy. Univ Geneva, Med Ctr, Dept Morphol, CH-1211 Geneva 4, Switzerland. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Taraboletti, G (reprint author), Mario Negri Inst Pharmacol Res, Dept Oncol, Via Gavazzeni 11, I-24125 Bergamo, Italy. RI Stetler-Stevenson, William/H-6956-2012; OI Stetler-Stevenson, William/0000-0002-5500-5808; Ghilardi, Carmen/0000-0002-3036-8557 NR 40 TC 34 Z9 37 U1 0 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD AUG 1 PY 2000 VL 258 IS 2 BP 384 EP 394 DI 10.1006/excr.2000.4936 PG 11 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 340JT UT WOS:000088529900016 PM 10896789 ER PT J AU Avichezer, D Chan, CC Silver, PB Wiggert, B Caspi, RR AF Avichezer, D Chan, CC Silver, PB Wiggert, B Caspi, RR TI Residues 1-20 of IRBP and whole IRBP elicit different uveitogenic and immunological responses in interferon gamma deficient mice SO EXPERIMENTAL EYE RESEARCH LA English DT Article DE autoimmune disease; interphotoreceptor retinoid binding-protein interferon-gamma; uveitis; cosinophils ID EXPERIMENTAL AUTOIMMUNE UVEORETINITIS; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; COLLAGEN-INDUCED ARTHRITIS; IFN-GAMMA; PROTECTIVE ROLE; CELL-FUNCTION; T-CELLS; SUSCEPTIBILITY; INDUCTION; DISEASE AB Experimental autoimmune uveoretinitis (EAU) is a T-cell-mediated autoimmune disease induced by immunization with uveitogenic retinal antigens, or by the adoptive transfer of uveitogenic T-cells of the Th-l-like phenotype. We have previously shown that IFN-gamma-deficient mice (GKO) on the C57BL/6 background are equally susceptible to interphotoreceptor retinoid binding protein (IRBP)-induced EAU as the wild type (WT). In the present study, we evaluated EAU induction in GKO mice by the newly described 11-2(b) epitope contained in residues 1-20 of human IRBP, and compared it to the response to the whole IRBP molecule. Similarly to previous observations with IRBP-induced EAU, delayed type hypersensitivity (DTH) and lymphocyte proliferation responses were elevated in GKO mice, as was production of IL-5 and TNF-alpha. However unlike the responses induced by whole IRBP, there was no delectable IL-IO production to the peptide. Histopathology on day 21 after immunization. revealed that both GKO and WT mice developed retinal lesions, including damage to the photoreceptor cell layer, vasculitis and inflammatory cellular infiltration, but disease scores were significantly higher in GKO, and retinal detachment was observed only in GKO mice. In contrast to the wild type, the cellular infiltrate in eyes of GKO mice contained a prominent component of eosinophils, although of lower proportion in peptide-induced than in IRBP-induced EAU. We conclude that the cytokine and inflammatory responses to human peptide 1-20 differ perceptibly from the responses to whole bovine IRBP, and may explain the elevated EAU scores of GKO mice compared to wild type. C1 NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. NEI, Lab Cell & Mol Biol, NIH, Bethesda, MD 20892 USA. RP Caspi, RR (reprint author), NEI, Immunol Lab, NIH, Bldg 10,Rm 10N222, Bethesda, MD 20892 USA. NR 32 TC 29 Z9 31 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0014-4835 J9 EXP EYE RES JI Exp. Eye Res. PD AUG PY 2000 VL 71 IS 2 BP 111 EP 118 DI 10.1006/exer.2000.0860 PG 8 WC Ophthalmology SC Ophthalmology GA 344BB UT WOS:000088736800001 PM 10930316 ER PT J AU Spector, A Zhou, W Ma, WC Chignell, CF Reszka, KJ AF Spector, A Zhou, W Ma, WC Chignell, CF Reszka, KJ TI Investigation of the mechanism of action of microperoxidase-11, (MP11), a potential anti-cataract agent, with hydrogen peroxide and ascorbate SO EXPERIMENTAL EYE RESEARCH LA English DT Article DE oxygen; oxidative stress; mass spectroscopy; electron paramagnetic resonance; lens ID HEME OCTAPEPTIDE MICROPEROXIDASE-8; LIPID-PEROXIDATION; CHELATED IRON; CYTOCHROME-C; H2O2; OXIDATION; DEHYDROASCORBATE; KINETICS; CATALASE; DEUTEROFERRIHEME AB The interaction of hydrogen peroxide, ascorbate and microperoxidase-ll (MP11), a ferriheme undecapeptide derived from cytochrome c, has been investigated using spectrophotometry, oxymetry, electron paramagnetic resonance (EPR), and mass spectroscopy techniques. It is shown that in 50 mM phosphate pH 7.0-7.4 in the absence of other reactants H2O2 induces a concentration-dependent decrease in absorption at the Soret band (399 nm) of the microperoxidase, with concomitant H2O2 decomposition and oxygen evolution. The reaction causes irreversible heme degradation, concomitant with loss of enzymatic activity. Ascorbate effectively protects MP11 from degradation and inhibits oxygen evolution. At ascorbate concentrations greater than that of H2O2, microperoxidase degradation is almost completely prevented. Mass spectrometry showed that H2O2 oxidizes the microperoxidase to a mono-oxygenated product, which did not form if ascorbate was included in the reaction system, There appears to be a 1:1 relationship between H2O2 degradation and ascorbate oxidation. EPR experiments revealed that an ascorbate radical was formed during the reaction, These reactions may be described by a scheme where a putative 'compound I' of the microperoxidase is reduced by ascorbate back to the original redox state (ferric) of the peroxidase in two one-electron steps, concomitantly with oxidation of the ascorbate to an ascorbate radical or in one two-electron transfer step forming dehydroascorbate. In the absence of ascorbate, the 'compound I' reacts further with the peroxide causing microperoxidase degradation and partial oxygen evolution. These observations are relevant to the interaction of ferrihemes with H2O2 and ascorbic acid and may he pertinent for the potential application of MP11 as an anti-cataract agent, (C) 2000 Academic Press. C1 Columbia Univ, Dept Ophthalmol, Biochem & Mol Biol Lab, New York, NY 10032 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. RP Spector, A (reprint author), Columbia Univ, Dept Ophthalmol, Biochem & Mol Biol Lab, 630 W 168th St, New York, NY 10032 USA. NR 39 TC 17 Z9 18 U1 2 U2 9 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0014-4835 J9 EXP EYE RES JI Exp. Eye Res. PD AUG PY 2000 VL 71 IS 2 BP 183 EP 194 DI 10.1006/exer.2000.0867 PG 12 WC Ophthalmology SC Ophthalmology GA 344BB UT WOS:000088736800008 PM 10930323 ER PT J AU Lane, MA AF Lane, MA TI Nonhuman primate models in biogerontology SO EXPERIMENTAL GERONTOLOGY LA English DT Review DE biogerontology; nonhuman primate models; calorie restriction ID FEMALE RHESUS-MONKEYS; CALORIE RESTRICTION; MACACA-MULATTA; RECOGNITION MEMORY; COGNITIVE DECLINE; DIABETES-MELLITUS; AGED MONKEY; DIETARY RESTRICTION; PIGTAILED MACAQUES; ALZHEIMERS-DISEASE AB A variety of animal models are utilized in biogerontological studies including yeast, nematodes, fruit flies, hamsters, mice, rats, and nonhuman primates. Species selection for research is based on many factors including economic feasibility, husbandry, generalizability of findings, available background information, adaptability to experimentation, and often, relevance to human aging. Each model offers its own strengths and limitations; however, nonhuman primates offer the unique advantage of phylogenetic proximity to humans. Among others, costs to purchase and maintain research subjects represent major limitations of nonhuman primate models. Although several nonhuman primate species have been utilized in aging research, rhesus monkeys (Macaca mulatta) are the best characterized and most extensively studied in biomedical gerontology. Nonhuman primate models have been employed as models for human aging in many research areas including neurobiology, skeletal, and reproductive aging and age-related diseases such as cardiovascular disease and diabetes. Primate models are now also being utilized to study interventions into aging such as caloric restriction. It will be several more years until definitive conclusions regarding lifespan effects can be made. However, existing data strongly suggest that many of the beneficial effects reported in rodents on CR also occur in primate models thereby strengthening the possibility that this nutritional paradigm may also impact favorably upon human aging. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NIA, Gerontol Res Ctr, Intramural Res Program, Baltimore, MD 21224 USA. RP Lane, MA (reprint author), NIA, Gerontol Res Ctr, Intramural Res Program, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 69 TC 41 Z9 43 U1 0 U2 4 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0531-5565 J9 EXP GERONTOL JI Exp. Gerontol. PD AUG PY 2000 VL 35 IS 5 BP 533 EP 541 DI 10.1016/S0531-5565(00)00102-9 PG 9 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 358TU UT WOS:000089573900003 PM 10978676 ER PT J AU Hong, SH Ondrey, FG Avis, IM Chen, Z Loukinova, E Cavanaugh, PF Van Waes, C Mulshine, JL AF Hong, SH Ondrey, FG Avis, IM Chen, Z Loukinova, E Cavanaugh, PF Van Waes, C Mulshine, JL TI Cyclooxygenase regulates human oropharyngeal carcinomas via the proinflammatory cytokine IL-6: a general role for inflammation? SO FASEB JOURNAL LA English DT Article DE colon cancer; arachidonic acid; inflammatory disease; COX inhibitors ID SQUAMOUS-CELL CARCINOMA; PROSTAGLANDIN E(2) PRODUCTION; HOST ENVIRONMENT PROMOTES; FACTOR-KAPPA-B; CANCER CELLS; ADULT PERIODONTITIS; TUMOR PROGRESSION; EPITHELIAL-CELLS; LUNG-CANCER; BONE LOSS AB High levels of prostaglandins are produced in human oropharyngeal carcinoma (OPC), Five human OPC cell lines tested expressed both isoforms of cyclooxygenases (COX), The pan-COX inhibitor ketorolac continuously and significantly decreased PGE(2) production and IL-6 and IL-8 levels in all OPC cell lines tested, but did not affect IL-1 alpha, GM-CSF levels, or in vitro tumor cell growth. In contrast, ketorolac reduced OPC growth in vivo. The OPC cell lines used express the IL-6 receptor, and IL-6 stimulation of these cells causes transduction to occur via STAT3 pathway activation. Coincubation with OPC cell lines with conditioned medium from a TPA-exposed HL-60 cells stimulated growth proportional to the IL-6 levels measured in the conditioned medium. This growth effect was specifically inhibited by anti-IL-6 antibody. These results are consistent with cytokine products of inflammatory cells having paracrine growth effects on OPC, If chronic inflammation plays a role in promoting the development of OPC, this mechanism may also apply to other epithelial tumor systems modulated by COX activity. C1 NCI, Med Branch, Div Clin Sci, Cell & Canc Biol Dept,Intervent Sect, Bethesda, MD 20892 USA. NIH, Natl Inst Deafness & Other Commun Disorders, Head & Neck Surg Branch, Bethesda, MD 20892 USA. Procter & Gamble Oral Hlth Care Technol Div, Cincinnati, OH USA. RP Mulshine, JL (reprint author), NCI, Med Branch, Div Clin Sci, Cell & Canc Biol Dept,Intervent Sect, Bldg 10-12N226,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 53 TC 64 Z9 69 U1 0 U2 2 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 2000 VL 14 IS 11 BP 1499 EP 1507 DI 10.1096/fj.14.11.1499 PG 9 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 342CN UT WOS:000088627800004 PM 10928984 ER PT J AU Mautino, MR Keiser, N Morgan, RA AF Mautino, MR Keiser, N Morgan, RA TI Improved titers of HIV-based lentiviral vectors using the SRV-1 constitutive transport element SO GENE THERAPY LA English DT Article DE rev-independent; lentiviral vector; CTE; gene therapy; HIV/AIDS ID IMMUNODEFICIENCY-VIRUS TYPE-1; GENE DELIVERY; REV FUNCTION; REPLICATION; REQUIREMENT; PROPAGATION; EXPRESSION; PROTEIN; RNA AB The development of lentiviral vectors that use Rev-independent mechanisms of nuclear export for their genomic RNA could facilitate the construction of novel anti-HIV vectors. We have improved the titers of Rev-independent lentiviral vectors having the SRV-1 CTE by mutating the major splice donor and acceptor sites present in the vector and by relocalization of the CTE sequences adjacent to the HIV-I 3'LTR. These two modifications have additive beneficial effects on vector titers and packaging efficiency. Packaging these CTE+ vectors expressing marker genes with a Rev-dependent HIV-1 helper vector yields higher titers than are obtained using a Rev-dependent lentiviral vector. C1 NHGRI, Clin Genet Therapy Branch, NIH, Bethesda, MD 20892 USA. RP Morgan, RA (reprint author), NHGRI, Clin Genet Therapy Branch, NIH, 10 Ctr Dr,Bldg 10,Room 10C103, Bethesda, MD 20892 USA. NR 17 TC 22 Z9 22 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0969-7128 J9 GENE THER JI Gene Ther. PD AUG PY 2000 VL 7 IS 16 BP 1421 EP 1424 DI 10.1038/sj.gt.3301262 PG 4 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 345UZ UT WOS:000088834700011 PM 10981670 ER PT J AU Flomerfelt, FA Kim, MG Schwartz, RH AF Flomerfelt, FA Kim, MG Schwartz, RH TI Spatial, a gene expressed in thymic stromal cells, depends on three-dimensional thymus organization for its expression SO GENES AND IMMUNITY LA English DT Article DE rodent; thymus; stromal cells; lymphoid organization; molecular biology ID MOUSE THYMUS; FETAL LIVER; IKAROS GENE; MICE; THYMOCYTES; SIGNALS; DIFFERENTIATION; REARRANGEMENT; HOMEOSTASIS; MATURATION AB Although the importance of thymic stroma in thymopoiesis has been recognized, the underlying molecular details regarding stromal cell biology remain obscure. To study this area, we have cloned genes expressed in thymic stromal cells. Spatial is alternatively spliced to generate two mRNAs in thymus and lymph node (LN) but it is not expressed in the spleen. In mouse embryos, the short form begins expression at day 10 while the long form is not detected until day 12. Both mRNAs encode proline rich proteins and their closest homology is to homeobox and POU domain transcription factors. Spatial is not expressed in thymocytes, but it is expressed in 2-deoxyguanosine-treated day 14 fetal thymic organ culture (FTOC) and in reaggregated FTOC. These data suggest that a normal three-dimensional organization of stromal cells is required for Spatial expression. An antiserum raised against a C-terminal peptide detected proteins of 38 and 32 kDa in Western blots of total thymus proteins. In frozen thymus sections, subcapsular epithelial cells were stained with the anti-Spatial antiserum. Paracortical subcapsular cells of unknown function were also stained in the LN. Both forms of Spatial fused to the green fluorescent protein (GFP) localize to the nucleus in transfected cells. C1 NIAID, Cellular & Mol Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Flomerfelt, FA (reprint author), NIAID, Cellular & Mol Immunol Lab, NIH, Bldg 10,Rm 12N266, Bethesda, MD 20892 USA. NR 41 TC 13 Z9 14 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1466-4879 J9 GENES IMMUN JI Genes Immun. PD AUG PY 2000 VL 1 IS 6 BP 391 EP 401 DI 10.1038/sj.gene.6363695 PG 11 WC Genetics & Heredity; Immunology SC Genetics & Heredity; Immunology GA 343HF UT WOS:000088695800008 PM 11196687 ER PT J AU Kaffer, CR Srivastava, M Park, KY Ives, E Hsieh, S Batlle, J Grinberg, A Huang, SP Pfeifer, K AF Kaffer, CR Srivastava, M Park, KY Ives, E Hsieh, S Batlle, J Grinberg, A Huang, SP Pfeifer, K TI A transcriptional insulator at the imprinted H19/Igf2 locus SO GENES & DEVELOPMENT LA English DT Article DE transcription epigenetics; chromatin; methylation; muscle ID MOUSE H19 GENE; METHYLATION IMPRINT; PRIMER EXTENSION; DROSOPHILA; EXPRESSION; IGF2; BOUNDARIES; ELEMENT; ALLELE; DOMAIN AB Igf2 and H19 exhibit parent-of-origin-specific monoallelic expression. H19 is expressed from the maternal chromosome and Igf2 from the paternal. The two genes share enhancer elements and monoallelic expression of both genes is dependent on cis-acting sequences upstream of the H19 promoter. In this work we examine the mechanisms by which this region silences the maternal Igf2 allele and we demonstrate that deletion of this region can result in high levels of activation of both H19 and Igf2 from a single chromosome. Moreover, by inserting this cis element between a promoter and its enhancer at a heterologous position, we demonstrate that the sequences carry both insulator activity and the ability to be stably imprinted. We also characterize the insulator in vitro and show that it is neither enhancer nor promoter specific. C1 NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. RP Pfeifer, K (reprint author), NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. OI Pfeifer, Karl/0000-0002-0254-682X NR 45 TC 120 Z9 122 U1 0 U2 1 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 0890-9369 J9 GENE DEV JI Genes Dev. PD AUG 1 PY 2000 VL 14 IS 15 BP 1908 EP 1919 PG 12 WC Cell Biology; Developmental Biology; Genetics & Heredity SC Cell Biology; Developmental Biology; Genetics & Heredity GA 342HX UT WOS:000088640100007 PM 10921905 ER PT J AU Kirchner, JM Tran, H Resnick, MA AF Kirchner, JM Tran, H Resnick, MA TI A DNA polymerase epsilon mutant that specifically causes +1 frameshift mutations within homonucleotide runs in yeast SO GENETICS LA English DT Article ID 5' EXONUCLEASE ACTIVITY; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; MISMATCH REPAIR; CATALYTIC SUBUNIT; STRUCTURAL ORGANIZATION; REPLICATION FIDELITY; I1307K POLYMORPHISM; MUTATOR PHENOTYPES; REPETITIVE DNA AB The DNA polymerases delta and epsilon are the major replicative polymerases in the yeast Saccharomyces cerevisiae that possess 3' --> 5' exonuclease proofreading activity. Many errors arising during replication are corrected by these exonuclease activities. We have investigated the contributions of regions of Pol epsilon other than thr proofreading motifs to replication accuracy. An allele, pol2-C1089Y, was identified in a screen of Pol epsilon mutants that in combination with an exonuclease I (exo1) mutation could cause a synergistic increase in mutations within homonucleotide runs. In contrast to other polymerase mutators, this allele specifically results in insertion frameshifts. When pol2-C1089Y was combined with deletions of EXO1 or RAD27 (homologue of human FEN1), mutation rates were increased for +1 frameshifts while there was almost no effect on -1 frame shifts. On the basis of genetic analysis, the pol2-C1089Y mutation did not cause a defect in proofreading. In combination with a deletion of the mismatch repair gene MSH2, the +1 frameshift mutation rate for a short homonucleotide run was increased nearly 100-fold whereas the -1 frameshift rate was unchanged. This suggests that the Pol2-C1089Y protein makes +1 frameshift errors during replication of homonucleotide mns and that these errors can be corrected by either mismatch repair (MMR) or proofreading (in short runs). This is the first report of a + 1-specific mutator for homonucleotide runs in vivo. The pol2-C1089Y mutation defines a functionally important residue in Pol epsilon. C1 NIEHS, Genet Mol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Resnick, MA (reprint author), NIEHS, Genet Mol Lab, NIH, Mail Drop D3-01,111 TW Alexander Dr,POB 12233, Res Triangle Pk, NC 27709 USA. NR 57 TC 24 Z9 24 U1 0 U2 0 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 USA SN 0016-6731 J9 GENETICS JI Genetics PD AUG PY 2000 VL 155 IS 4 BP 1623 EP 1632 PG 10 WC Genetics & Heredity SC Genetics & Heredity GA 342UX UT WOS:000088664500011 PM 10924461 ER PT J AU Aravind, L AF Aravind, L TI Guilt by association: Contextual information in genome analysis SO GENOME RESEARCH LA English DT Editorial Material ID PROTEIN FUNCTION; GENE ORDER; EVOLUTION 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. NR 19 TC 93 Z9 96 U1 0 U2 3 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 2000 VL 10 IS 8 BP 1074 EP 1077 DI 10.1101/gr.10.8.1074 PG 4 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 347ZC UT WOS:000088958200003 PM 10958625 ER PT J AU Kim, J Noskov, VN Li, XC Bergmann, A Ren, XJ Warth, T Richardson, P Kouprina, N Stubbs, L AF Kim, J Noskov, VN Li, XC Bergmann, A Ren, XJ Warth, T Richardson, P Kouprina, N Stubbs, L TI Discovery of a novel, paternally expressed ubiquitin-specific processing protease gene through comparative analysis of an imprinted region of mouse chromosome 7 and human chromosome 19q13.4 SO GENOME RESEARCH LA English DT Article ID BECKWITH-WIEDEMANN-SYNDROME; ZINC-FINGER PROTEIN; PROXIMAL CHROMOSOME-7; EVOLUTIONARY CHANGES; MATERNAL-BEHAVIOR; PRADER-WILLI; TRANSCRIPTS; PEG3; AMPLIFICATION; MUTATIONS AB Using mouse BAC clones spanning an imprinted interval of proximal mouse chromosome 7 and the genomic sequence of the related interval of human chromosome 19q13.4, we have identified a novel mouse gene, Usp29 (ubiquitin-specific processing protease 29), near two known imprinted genes, Peg3 and Ziml. Gene Usp29 is located directly adjacent to Peg3 in a "head-to-head" orientation, and comprises exons distributed over a genomic distance of at least 400 kb. A similar human gene is also Found in the homologous location in human chromosome 19q13.4. The mouse Usp29 gene is also imprinted and is transcribed mainly from the paternal allele with highest expression levels in adult brain, especially in the cerebral cortex and hippocampus, and in the Forebrain, face, and limb buds of midgestation mouse embryos. Analysis of a full-length 7.6-kb cDNA clone revealed that Usp29 encodes an 869-amino-acid protein that displays significant homology with yeast and nematode ubiquitin carboxyl-terminal hydrolases. These data suggest that, like the candidate Angelman syndrome gene Ube3a (ubiquitin ligase), Usp29 may represent another imprinted gene involved in the ubiquitination pathway. This identification of a third imprinted gene, Usp29, From the Peg3/Ziml-region confirms the presence of a conserved imprinted domain spanning at least 500 kb in the proximal portion of mouse chromosome 7 (Mmu7). C1 Univ Calif Lawrence Livermore Natl Lab, Ctr Human Genome, Biol & Biotechnol Res Program, Livermore, CA 94551 USA. DOE Joint Genome Inst, Walnut Creek, CA 94598 USA. NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. RP Stubbs, L (reprint author), Univ Calif Lawrence Livermore Natl Lab, Ctr Human Genome, Biol & Biotechnol Res Program, L-441, Livermore, CA 94551 USA. OI Stubbs, Lisa/0000-0002-9556-1972 NR 39 TC 51 Z9 56 U1 0 U2 2 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 2000 VL 10 IS 8 BP 1138 EP 1147 DI 10.1101/gr.10.8.1138 PG 10 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 347ZC UT WOS:000088958200010 PM 10958632 ER PT J AU Aravind, L Koonin, EV AF Aravind, L Koonin, EV TI Eukaryote-specific domains in translation initiation factors: Implications for translation regulation and evolution of the translation system SO GENOME RESEARCH LA English DT Article ID YEAST SACCHAROMYCES-CEREVISIAE; PROTEIN SECONDARY STRUCTURE; HORIZONTAL GENE-TRANSFER; PROGRAMMED CELL-DEATH; FACTOR 4G EIF4G; MESSENGER-RNA; BINDING PROTEIN; ANGSTROM RESOLUTION; FUNCTIONAL DOMAINS; FACTOR 5A AB Computational analysis of sequences of proteins involved in translation initiation in eukaryotes reveals a number of specific domains that are not represented in bacteria or archaea. Most of these eukaryote-specific domains are known or predicted to possess an alpha-helical structure, which suggests that such domains are easier to invent in the course of evolution than are domains of other structural classes. A previously undetected, conserved region predicted to form an alpha-helical domain is delineated in the initiation factor eIF4G, in Nonsense-mediated mRNA decay 2 protein (NMD2/UPF2), in the nuclear cap-binding CBP80, and in other, poorly characterized proteins, which is named the NIC (NMD2, eIF4G, CBP80) domain. Biochemical and mutagenesis data on NIC-containing proteins indicate that this predicted domain is one of the central adapters in the regulation of mRNA processing, translation, and degradation. it is demonstrated that, in the course of eukaryotic evolution, initiation factor eIF4G, of which NIC is the core, conserved portion, has accreted several additional, distinct predicted domains such as MI (MA-3 and eIF4G) and W2, which probably was accompanied by acquisition of new regulatory interactions. 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. NR 60 TC 62 Z9 66 U1 0 U2 1 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 2000 VL 10 IS 8 BP 1172 EP 1184 DI 10.1101/gr.10.8.1172 PG 13 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 347ZC UT WOS:000088958200013 PM 10958635 ER PT J AU Clifford, R Edmonson, M Hu, Y Nguyen, C Scherpbier, T Buetow, KH AF Clifford, R Edmonson, M Hu, Y Nguyen, C Scherpbier, T Buetow, KH TI Expression-based genetic/physical maps of single-nucleotide polymorphisms identified by the cancer genome anatomy project SO GENOME RESEARCH LA English DT Article ID RADIATION HYBRID MAP; COMPLEX TRAITS; GENETIC-MAP; LINKAGE; SNPS AB SNPs (Single-Nucleotide Polymorphisms), the most common DNA variant in humans, represent a valuable resource for the genetic analysis of cancer and other illnesses. These markers may be used in a variety of ways to investigate the genetic underpinnings of disease. In gene-based studies, the correlations between allelic variants of genes of interest and particular disease states are assessed. An extensive collection of SNP markers may enable entire molecular pathways regulating cell metabolism, growth, or differentiation to be analyzed by this approach. In addition, high-resolution genetic maps based on SNPs will greatly facilitate linkage analysis and positional cloning. The National Cancer Institute's CGAP-GAI (Cancer Genome Anatomy Project Genetic Annotation Initiative) group has identified 10,243 SNPs by examining publicly available EST (Expressed Sequence Tag) chromatograms. More than 6800 of these polymorphisms have been placed on expression-based integrated genetic/physical maps. in addition to a set of comprehensive SNP maps, we have produced maps containing single nucleotide polymorphisms in genes expressed in breast, colon, kidney, liver, lung, or prostate tissue. The integrated maps, a SNP search engine, and a lava-based tool for viewing candidate SNPs in the context of EST assemblies can be accessed via the CGAP-GAI web site [http://cgap.nci.nih.gov/GAI/). Our SNP detection tools are available to the public for noncommercial use. C1 NCI, Lab Populat Genet, NIH, Bethesda, MD 20892 USA. RP Buetow, KH (reprint author), NCI, Lab Populat Genet, NIH, Bethesda, MD 20892 USA. NR 29 TC 36 Z9 42 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 2000 VL 10 IS 8 BP 1259 EP 1265 DI 10.1101/gr.10.8.1259 PG 7 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 347ZC UT WOS:000088958200022 PM 10958644 ER PT J AU Teresi, JA Holmes, D Ory, MG AF Teresi, JA Holmes, D Ory, MG TI The therapeutic design of environments for people with dementia: Further reflections and recent findings from the National Institute on Aging collaborative studies of Dementia Special Care Units SO GERONTOLOGIST LA English DT Editorial Material ID ALZHEIMERS-DISEASE; ELDERLY SUBJECTS; NURSING-HOME; BEHAVIOR; LIGHT C1 Hebrew Home Aged, Div Res, Riverdale, NY 10471 USA. New York State Psychiat Inst, New York, NY 10032 USA. Columbia Univ, Stroud Ctr, New York, NY USA. NIA, Bethesda, MD 20892 USA. RP Teresi, JA (reprint author), Hebrew Home Aged, Div Res, 5901 Palisade Ave, Riverdale, NY 10471 USA. FU NIA NIH HHS [UO1AG10330] NR 26 TC 15 Z9 15 U1 0 U2 9 PU GERONTOLOGICAL SOCIETY AMER PI WASHINGTON PA 1275 K STREET NW SUITE 350, WASHINGTON, DC 20005-4006 USA SN 0016-9013 J9 GERONTOLOGIST JI Gerontologist PD AUG PY 2000 VL 40 IS 4 BP 417 EP 421 PG 5 WC Gerontology SC Geriatrics & Gerontology GA 373VP UT WOS:000165311000003 PM 10961030 ER PT J AU Schlemper, RJ Riddell, RH Kato, Y Borchard, F Cooper, HS Dawsey, SM Dixon, MF Fenoglio-Preiser, CM Flejou, JF Geboes, K Hattori, T Hirota, T Itabashi, M Iwafuchi, M Iwashita, A Kim, YI Kirchner, T Klimpfinger, M Koike, M Lauwers, GY Lewin, KJ Oberhuber, G Offner, F Price, AB Rubio, CA Shimizu, M Shimoda, T Sipponen, P Solcia, E Stolte, M Watanabe, H Yamabe, H AF Schlemper, RJ Riddell, RH Kato, Y Borchard, F Cooper, HS Dawsey, SM Dixon, MF Fenoglio-Preiser, CM Flejou, JF Geboes, K Hattori, T Hirota, T Itabashi, M Iwafuchi, M Iwashita, A Kim, YI Kirchner, T Klimpfinger, M Koike, M Lauwers, GY Lewin, KJ Oberhuber, G Offner, F Price, AB Rubio, CA Shimizu, M Shimoda, T Sipponen, P Solcia, E Stolte, M Watanabe, H Yamabe, H TI The Vienna classification of gastrointestinal epithelial neoplasia SO GUT LA English DT Article DE early carcinoma; adenoma; dysplasia; oesophagus; stomach; colon ID WESTERN PATHOLOGISTS; DIAGNOSTIC-CRITERIA; CARCINOMA; JAPANESE AB Background-Use of the conventional Western and Japanese classification systems of gastrointestinal epithelial neoplasia results in large differences among pathologists in the diagnosis of oesophageal, gastric, and colorectal neoplastic lesions. Aim-To develop common worldwide terminology for gastrointestinal epithelial neoplasia. Methods-Thirty one pathologists from 12 countries reviewed 35 gastric, 20 colorectal, and 21 oesophageal biopsy and resection specimens. The extent of diagnostic agreement between those with Western and Japanese viewpoints was assessed by kappa statistics. The pathologists met in Vienna to discuss the results and to develop a new consensus terminology. Results-The large differences between the conventional Western and Japanese diagnoses were confirmed (percentage of specimens for which there was agreement and kappa values: 37% and 0.16 for gastric; 45% and 0.27 for colorectal; and 14% and 0.01 for oesophageal lesions). There was much better agreement among pathologists (71% and 0.55 for gastric; 65% and 0.47 for colorectal; and 62% and 0.31 for oesophageal lesions) when the original assessments of the specimens were regrouped into the categories of the proposed Vienna classification of gastrointestinal epithelial neoplasia: (1) negative for neoplasia/dysplasia, (2) indefinite for neoplasia/dysplasia, (3) non-invasive low grade neoplasia (low grade adenoma/dysplasia), (4) non-invasive high grade neoplasia (high grade adenoma/dysplasia, non-invasive carcinoma and suspicion of invasive carcinoma), and (5) invasive neoplasia (intramucosal carcinoma, submucosal carcinoma or beyond). Conclusion-The differences between Western and Japanese pathologists in the diagnostic classification of gastrointestinal epithelial neoplastic lesions can be resolved largely by adopting the proposed terminology, which is based on cytological and architectural severity and invasion status. C1 Fukuoka Univ, Sch Med, Dept Internal Med, Jonan Ku, Fukuoka 8140180, Japan. McMaster Univ, Med Ctr, Dept Pathol & Mol Med, Hamilton, ON, Canada. Fox Chase Canc Ctr, Dept Pathol, Philadelphia, PA 19111 USA. NCI, Div Clin Sci, Bethesda, MD 20892 USA. Univ Leeds, Ctr Digest Dis, Dept Histopathol, Leeds, W Yorkshire, England. Univ Cincinnati, Med Ctr, Dept Pathol & Lab Med, Cincinnati, OH 45267 USA. St Antonius Hosp, Serv Anat Pathol, Paris, France. Katholieke Univ Leuven Hosp, Dept Pathol, Louvain, Belgium. Shiga Univ Med Sci, Dept Pathol, Otsu, Shiga 52021, Japan. Special Reference Lab, Tokyo, Japan. Ibaraki Cent Hosp, Dept Pathol, Tomobe, Japan. Niigata Univ, Dept Med Technol, Niigata, Japan. Fukuoka Univ, Chikushi Hosp, Dept Pathol, Fukuoka 81401, Japan. Seoul Natl Univ, Dept Pathol, Seoul 151, South Korea. Univ Erlangen Nurnberg, Inst Pathol, D-8520 Erlangen, Germany. Kaiser Franz Josef Spital, Inst Pathol & Microbiol, Vienna, Austria. Tokyo Metropolitan Komagome Hosp, Dept Pathol, Tokyo, Japan. Univ Florida, Hlth Sci Ctr, Dept Pathol, Gainesville, FL USA. Univ Calif Los Angeles, Ctr Hlth Sci, Dept Pathol & Lab Med, Los Angeles, CA 90024 USA. Univ Vienna, Dept Clin Pathol, Vienna, Austria. Univ Innsbruck, Dept Pathol, A-6020 Innsbruck, Austria. Karolinska Inst, Gastrointestinal Liver Pathol Res Lab, Stockholm, Sweden. Hokkaido Univ Hosp, Dept Surg Pathol, Sapporo, Hokkaido 060, Japan. Natl Canc Ctr, Clin Lab Div, Tokyo, Japan. Jorvi Hosp, Dept Pathol, SF-02740 Espoo, Finland. Univ Pavia, Dept Human Pathol, I-27100 Pavia, Italy. Policlin San Matteo, I-27100 Pavia, Italy. Klinikum Bayreuth, Inst Pathol, Bayreuth, Germany. Niigata Univ, Sch Med, Dept Pathol 1, Niigata, Japan. Kyoto Univ Hosp, Anat Pathol Lab, Kyoto 606, Japan. RP Schlemper, RJ (reprint author), Fukuoka Univ, Sch Med, Dept Internal Med, Jonan Ku, 7-45-1 Nanakuma, Fukuoka 8140180, Japan. EM ronald-s@fukuoka-u.ac.jp RI Riddell, Robert/J-5557-2016 NR 3 TC 1041 Z9 1093 U1 0 U2 23 PU B M J PUBLISHING GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0017-5749 J9 GUT JI Gut PD AUG PY 2000 VL 47 IS 2 BP 251 EP 255 DI 10.1136/gut.47.2.251 PG 5 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 337UU UT WOS:000088380700018 PM 10896917 ER PT J AU Farley, J Gray, K Nycum, L Prentice, M Birrer, MJ Jakowlew, SB AF Farley, J Gray, K Nycum, L Prentice, M Birrer, MJ Jakowlew, SB TI Endocervical cancer is associated with an increase in the ligands and receptors for transforming growth factor-beta and a contrasting decrease in p27(Kip1) SO GYNECOLOGIC ONCOLOGY LA English DT Article DE TGF-beta; p27(Kip1); endocervix; carcinogenesis ID SQUAMOUS INTRAEPITHELIAL NEOPLASIA; COLON-CARCINOMA-CELLS; HUMAN BREAST-CANCER; TGF-BETA; UTERINE CERVIX; II RECEPTOR; ADENOCARCINOMA INSITU; C-MYC; MICROSATELLITE INSTABILITY; ALTERED EXPRESSION AB Objective. The aim of this study was to investigate the relationship between the expression of the TGF-beta ligands and TGF-beta receptors to the expression of p27(Kip1), a TGF-beta-regulated gene, in endocervical cancer, Methods. To examine the expression of TGF-beta and p27(Kip1) in malignant transformation of the uterine endocervix, a panel of 23 formalin-fixed and paraffin-embedded human cervical specimens, including 8 with benign endocervical glands, 8 with cervical adenocarcinoma in situ, and 7 with cervical adenocarcinomas, was used. Tissues were immunostained with polyclonal antibodies that react specifically with TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta RI, TGF-beta RII, and p27(Kip1). Results. Immunostaining for TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta RI, TGF-beta RII, and p27(Kip1) was detected in normal endocervix, with the TGF-beta s showing weak cytoplasmic staining, while p27(Kip1) showed strong nuclear staining, Expression of TGF-beta increased significantly upon neoplastic transformation with the TGF-beta ligands and receptors showing strong cytoplasmic staining in adenocarcinoma in situ compared to normal endocervix, Interestingly, expression of TGF-beta was lower in adenocarcinoma than in adenocarcinoma in situ, but still significantly higher than in normal endocervix, TGF-beta 2 and TGF-beta 3 showed higher levels of immunostaining than TGF-beta 1 in adenocarcinomas, In contrast, p27(Kip1) protein expression decreased with progressive malignancy, with lower p27(Kip1) protein levels detected in adenocarcinoma than in adenocarcinoma in situ, while normal endocervix showed the highest level of p27(Kip1) protein expression. Conclusion. Elevated expression of the TGF-beta ligands and receptors is found in both cervical adenocarcinoma in situ and adenocarcinoma compared to normal endocervix, In contrast, a progressive decrease in p27(Kip1) occurs upon neoplastic transformation of the normal endocervix to cervical adenocarcinoma, These results suggest that neoplastic transformation of the endocervix may be related to dysregulation of TGF-beta and p27(Kip1) seen as an elevation of TGF-beta and a reduction of p27(Kip1) expression that may lead to loss of cell cycle control. C1 NCI, Dept Cell & Canc Biol, Med Branch, Div Clin Sci, Rockville, MD 20850 USA. Walter Reed Army Med Ctr, Dept Gynecol Oncol, Washington, DC 20307 USA. Uniformed Serv Univ Hlth Sci, Dept Obstet & Gynecol, Bethesda, MD 20814 USA. Naval Med Ctr Portsmouth, Dept Obstet & Gynecol, Div Gynecol Oncol, Norfolk, VA 23505 USA. RP Jakowlew, SB (reprint author), NCI, Dept Cell & Canc Biol, Med Branch, Div Clin Sci, 9610 Med Ctr Dr,Suite 300, Rockville, MD 20850 USA. NR 73 TC 16 Z9 18 U1 0 U2 0 PU ACADEMIC PRESS INC 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 2000 VL 78 IS 2 BP 113 EP 122 DI 10.1006/gyno.2000.5879 PG 10 WC Oncology; Obstetrics & Gynecology SC Oncology; Obstetrics & Gynecology GA 345JJ UT WOS:000088812600006 PM 10926789 ER PT J AU Allegra, CJ Kramer, BS AF Allegra, CJ Kramer, BS TI Understanding clinical trials - Preface SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Editorial Material C1 NCI, NIH, Bethesda, MD 20892 USA. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Allegra, CJ (reprint author), NCI, NIH, 10 Ctr Dr,MSC 1906,Bldg 10,Room 12N226, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP XIII EP XIV DI 10.1016/S0889-8588(05)70308-7 PG 2 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300001 ER PT J AU Meldrum, ML AF Meldrum, ML TI A brief history of the randomized controlled trial - From oranges and lemons to the gold standard SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID SURGICAL-ADJUVANT-BREAST; CANCER PREVENTION TRIAL; CLINICAL-TRIAL; ACTIVISM AB This article discusses the history and development of randomized clinical trial methodology, the reasons for its status and authority as a method of therapeutic evaluation, and the continuing role of clinical judgement in designing, interpreting, and applying the findings of trials. C1 Natl Inst Dent & Craniofacial Res, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD USA. RP Meldrum, ML (reprint author), 18311 Lost Knife Circle 202, Montgomery Village, MD 20886 USA. NR 51 TC 37 Z9 39 U1 8 U2 17 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 745 EP + DI 10.1016/S0889-8588(05)70309-9 PG 17 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300002 PM 10949771 ER PT J AU Weed, DL AF Weed, DL TI Epidemiologic evidence and causal inference SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID WEAK ASSOCIATIONS; METAANALYSIS; CANCER; GUIDELINES; CRITERIA AB Preventing cancer depends on the ability to recognize and remove causal factors. In current practice, the methods used to judge cause from epidemiologic, clinical trials and biologic evidence include systematic narrative reviews, criteria-based inference methods, and meta-analysis. Subjectivity and values play a key role in the practice of causal inference, especially in selecting criteria and assigning rules of evidence to those criteria. Judging cause is a central concern of physicians, epidemiologists, and other public health professionals committed to cancer prevention. C1 NCI, Off Prevent Oncol, Div Canc Prevent, Bethesda, MD 20892 USA. Georgetown Univ, Kennedy Inst Eth, Washington, DC 20057 USA. RP Weed, DL (reprint author), NCI, Off Prevent Oncol, Div Canc Prevent, 6130 Execut Blvd,Suite T-41, Bethesda, MD 20892 USA. NR 41 TC 14 Z9 14 U1 2 U2 4 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 797 EP + DI 10.1016/S0889-8588(05)70312-9 PG 12 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300005 PM 10949774 ER PT J AU Hawk, ET Lippman, SM AF Hawk, ET Lippman, SM TI Primary cancer prevention trials SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Review ID CORONARY HEART-DISEASE; SURGICAL-ADJUVANT-BREAST; SURROGATE END-POINTS; NONSTEROIDAL ANTIINFLAMMATORY DRUGS; RANDOMIZED CONTROLLED TRIAL; BOWEL-PROJECT P-1; BETA-CAROTENE; CARDIOVASCULAR-DISEASE; COLORECTAL-CANCER; CLINICAL-TRIAL AB Cancer, formerly considered a clinically catastrophic event, is but one late stage in a long disease process driven by cumulative molecular alterations that affect genomic integrity and control of cellular identity and growth. improved understanding of this process has led to opportunities for earlier intervention, thereby preventing invasive cancers. Approval of an agent for breast cancer risk reduction has raised professional and public expectations for preventive oncology and fostered excitement in this rapidly growing field. This article examines the current status of, and future prospects for, the science and practice of cancer chemoprevention. C1 Univ Texas, MD Anderson Canc Ctr, Dept Clin Canc Prevent, Houston, TX 77030 USA. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Lippman, SM (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Clin Canc Prevent, 1515 Holcombe Blvd,Box 236, Houston, TX 77030 USA. FU NCI NIH HHS [CA16672] NR 109 TC 3 Z9 3 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 809 EP + DI 10.1016/S0889-8588(05)70313-0 PG 23 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300006 PM 10949775 ER PT J AU Kramer, BS Brawley, OW AF Kramer, BS Brawley, OW TI Cancer screening SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID AGE-SPECIFIC INCIDENCE; SURROGATE END-POINTS; LUNG-CANCER; CLINICAL-TRIALS; NEUROBLASTOMA; BENEFITS; JAPAN AB The practicing physician is faced with multiple cancer screening recommendations and guidelines from a variety of organizations. These organizations often have different constituencies and perspectives, resulting in conflicting guidelines. This article is designed to help physicians assess the strength of evidence supporting screening tests in order to help inform discussion with patients about undergoing screening. C1 NCI, Div Canc Prevent, Bethesda, MD 20892 USA. NCI, Off Special Populat Res, Bethesda, MD 20892 USA. NCI, Off Director, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Dept Med, Bethesda, MD 20814 USA. RP Kramer, BS (reprint author), NCI, Div Canc Prevent, 31 Ctr Dr,Room 10A49,MSC 2580, Bethesda, MD 20892 USA. NR 26 TC 18 Z9 18 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 831 EP + DI 10.1016/S0889-8588(05)70314-2 PG 19 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300007 PM 10949776 ER PT J AU Rubinstein, LV AF Rubinstein, LV TI Therapeutic studies SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID II CLINICAL-TRIALS; MULTIPLE TESTING PROCEDURE; CONFIDENCE-INTERVALS; DESIGN; CANCER; GUIDELINES; TOXICITY AB This article discusses the problems in basic design, conduct, and interpretation associated with phases I, II, and III of the cancer clinical trials and explains the various statistical solutions to these problems. The fundamental problem common to all three trials is achieving a correct and precise answer to the question posed to inform future testing and treatment while protecting trial patients from receiving treatment that has demonstrated excessive toxicity or lack of clinical efficacy. This shared problem gives rise to statistical designs with basic similarities across the three trial types. C1 NCI, Biometr Res Branch, Bethesda, MD 20892 USA. RP Rubinstein, LV (reprint author), NCI, Biometr Res Branch, EPN 739,6120 Execut Blvd MSC 7434, Bethesda, MD 20892 USA. NR 54 TC 8 Z9 9 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 849 EP + DI 10.1016/S0889-8588(05)70315-4 PG 29 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300008 PM 10949777 ER PT J AU Schatzkin, A AF Schatzkin, A TI Intermediate markers as surrogate endpoints in cancer research SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID HUMAN PAPILLOMAVIRUS INFECTION; RECTAL MUCOSAL PROLIFERATION; CERVICAL INTRAEPITHELIAL NEOPLASIA; RANDOMIZED CONTROLLED TRIAL; CELL NUCLEAR ANTIGEN; END-POINTS; POSTMENOPAUSAL WOMEN; COLORECTAL-CANCER; CLINICAL-TRIALS; VALIDATION AB Studies employing surrogate cancer endpoints can be smaller, shorter, and less expensive than studies with malignant disease endpoints. A broad range of histologic, cellular, and molecular markers may serve as surrogate endpoints for cancer. An understanding of the causal structure underlying the relations of exposures, potential surrogate endpoints, and cancer is critical in evaluating surrogates. Three elements pertinent to elucidating this structure are: (1) the relation of the surrogate endpoint to cancer; (2) the relation of the intervention (exposure) to the surrogate; and (3) the extent to which the surrogate endpoint mediates the relation between the intervention and cancer. C1 NCI, Div Canc Epidemiol & Genet, Nutr Epidemiol Branch, Bethesda, MD 20892 USA. RP Schatzkin, A (reprint author), NCI, Div Canc Epidemiol & Genet, Nutr Epidemiol Branch, 6120 Execut Blvd,EPS 7032, Bethesda, MD 20892 USA. NR 40 TC 8 Z9 9 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 887 EP + DI 10.1016/S0889-8588(05)70317-8 PG 20 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300010 PM 10949779 ER PT J AU Misra, RR Pinsky, PF Srivastava, S AF Misra, RR Pinsky, PF Srivastava, S TI Prognostic factors for hematologic cancers SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID ARTIFICIAL NEURAL NETWORKS; MALIGNANT MYELOID DISEASES; LOGISTIC-REGRESSION; MULTIPLE-MYELOMA; MOLECULAR DELINEATION; CROSS-VALIDATION; GENE-EXPRESSION; GENOMIC-SCALE; CELL LEUKEMIA; TAX PROTEIN AB This article reviews the molecular biology of hematologic cancers and the current understanding of prognostic factors for these cancers. Specific molecular biomarkers that have potential as prognostic factors for various hematologic cancers are discussed. Quantitative and statistical methods of evaluating the usefulness of prognostic factors are presented. C1 NCI, Div Canc Prevent, Canc Biomarkers Res Grp, Bethesda, MD 20892 USA. NCI, Early Detect Res Grp, Canc Prevent Fellowship Program, Bethesda, MD 20892 USA. RP Srivastava, S (reprint author), NCI, Div Canc Prevent, Canc Biomarkers Res Grp, 6130 Execut Blvd,Suite 330F, Bethesda, MD 20892 USA. NR 65 TC 9 Z9 10 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 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD AUG PY 2000 VL 14 IS 4 BP 907 EP + DI 10.1016/S0889-8588(05)70318-X PG 20 WC Oncology; Hematology SC Oncology; Hematology GA 345DC UT WOS:000088799300011 PM 10949780 ER PT J AU Sakata, H Rubin, JS Taylor, WG Miki, T AF Sakata, H Rubin, JS Taylor, WG Miki, T TI A Rho-specific exchange factor Ect2 is induced from S to M phases in regenerating mouse liver SO HEPATOLOGY LA English DT Article ID HEPATOCYTE GROWTH-FACTOR; EXPRESSION CDNA CLONING; IN-SITU HYBRIDIZATION; TUMOR-NECROSIS-FACTOR; RAT-LIVER; PARTIAL-HEPATECTOMY; MESSENGER-RNA; HEPATIC REGENERATION; HUMAN ONCOGENE; FACTOR-ALPHA AB The ect2 oncogene was originally identified as a transforming complementary DNA (cDNA) from mouse epithelial cells in an expression cloning approach and encodes a product related to Rho-specific exchange factors and yeast cell cycle regulators. To explore the potential role of ect2 in the cell cycle, we examined the expression of the ect2 protooncogene in a liver regeneration model in mice after partial (two thirds) hepatectomy. We found that the expression of the ect2 transcript and protein were markedly elevated with the onset of DNA synthesis and remained elevated during G2 and M phases. The timing of cct2 expression matched that of proliferating cell nuclear antigen (PCNA) and partially overlapped cell division cycle 2 (Cdc2) expression. In situ hybridization analysis showed that cct2 was expressed at a high level in cells undergoing mitosis in regenerating liver. Moreover, expression of a dominant negative or an oncogenic mutant of cct2 in cultured mouse hepatocytes resulted in a large increase in the number of binucleated cells. These findings showed that Ect2 is expressed in a cell cycle-dependent manner during liver regeneration, and suggest that it has an important role in the regulation of cytokinesis. C1 NCI, Mol Tumor Biol Sect, Basic Res Lab, Bethesda, MD 20892 USA. NCI, Mol & Cellular Biol Lab, Bethesda, MD 20892 USA. RP Miki, T (reprint author), NCI, Mol Tumor Biol Sect, Basic Res Lab, Bldg 37,Room 1E24,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 57 TC 23 Z9 23 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 2000 VL 32 IS 2 BP 193 EP 199 DI 10.1053/jhep.2000.8271 PG 7 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 350FR UT WOS:000089090700004 PM 10915723 ER PT J AU Terai, S Aoki, H Ashida, K Thorgeirsson, SS AF Terai, S Aoki, H Ashida, K Thorgeirsson, SS TI Human homologue of maid: A dominant inhibitory helix-loop-helix protein associated with liver-specific gene expression SO HEPATOLOGY LA English DT Article ID OVAL CELL-DIFFERENTIATION; RAT SERUM-ALBUMIN; DNA-BINDING; TRANSCRIPTION FACTOR; NEGATIVE REGULATOR; ALPHA-FETOPROTEIN; ID PROTEINS; ENHANCER; SEQUENCE; DROSOPHILA AB The helix-loop-helix (HLH) family of transcriptional. regulatory proteins are key regulators in numerous developmental processes. The class I HLH proteins, such as E12 are ubiquitously expressed. Class II HLH proteins, such as MyoD, are expressed in a tissue-specific manner, Class I and II heterodimers can bind to E-boxes (CANNTG) and regulate lineage commitments of embryonic cells. In an attempt to identify partners for the E12 protein that may exert control during liver development, we performed the yeast 2-hybrid screen using an expression complementary DNA library from human fetal liver. A novel dominant inhibitory HLH factor, designated HHM (human homologue of maid), was isolated and characterized. HHM is structurally related to the Id family and was highly expressed in brain, pituitary gland, lung, heart, placenta, fetal liver, and bone marrow. HHM physically interacted with E12 in vitro and in mammalian cells. Comparison of the dominant inhibitory effects of HHM and Id2 on the binding of E12/MyoD dimer to an E-box element revealed a weaker inhibition by HHM. However, HHM but not Id2 specifically inhibited the luciferase gene activation induced by hepatic nuclear factor 4 (HNF4) promoter. The HHM was transiently expressed during stem-cell-driven regeneration of the liver at the stage in which the early basophilic foci of hepatocytes started to appear. These results suggest that HHM is a novel type of dominant inhibitory HLH protein that might modulate liver-specific gene expression. C1 NCI, Div Basic Sci, Expt Carcinogenesis Lab, Bethesda, MD 20892 USA. RP Thorgeirsson, SS (reprint author), NCI, Div Basic Sci, Expt Carcinogenesis Lab, Bldg 37,Room 3C28,37 Convent Dr MSC4255, Bethesda, MD 20892 USA. NR 61 TC 22 Z9 24 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 0270-9139 J9 HEPATOLOGY JI Hepatology PD AUG PY 2000 VL 32 IS 2 BP 357 EP 366 DI 10.1053/jhep.2000.9092 PG 10 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 350FR UT WOS:000089090700024 PM 10915743 ER PT J AU Tanaka, E Ohue, C Aoyagi, K Yamaguchi, K Yagi, S Kiyosawa, K Alter, HJ AF Tanaka, E Ohue, C Aoyagi, K Yamaguchi, K Yagi, S Kiyosawa, K Alter, HJ TI Evaluation of a new enzyme immunoassay for hepatitis C virus (HCV) core antigen with clinical sensitivity approximating that of genomic amplification of HCV RNA SO HEPATOLOGY LA English DT Article ID POLYMERASE CHAIN-REACTION; NON-B-HEPATITIS; NON-A; POSTTRANSFUSION HEPATITIS; INTERFERON-ALFA; SERUM; 2ND-GENERATION; TRANSFUSION; INFECTION; PROTEIN AB The aim of this study was to analyze the clinical performance of a new enzyme immunoassay (EIA) for hepatitis C virus (HCV) core antigen in comparison with the reverse transcription polymerase chain reaction (RT-PCR). A total of 310 patients with acute or chronic hepatitis C, and 132 HCV-negative controls were studied. Chemiluminescence EIA with monoclonal anti-HCV core antigen was used, and qualitative and quantitative commercial RT-PCRs and an in-house nested RT-PCR were performed. Compared with nested RT-PCR, the core antigen assay showed 97% sensitivity and 100% specificity in 75 patients with chronic hepatitis C and 132 controls. HCV core antigen was positive in 16 (94%) of 17 patients with acute hepatitis C at initial consultation. In 3 persons prospectively followed, core antigen was detected in the first available (1-3 weeks) post-transfusion sample. In 167 anti-HCV-positive individuals, 129 (77%) were viremic; core antigen was detected in 126 (98%) compared with 129 (100%) for nested RT-PCR and 121 (94%) for the commercial RT-PCR. In 48 patients with chronic hepatitis C treated with interferon alfa, the concentration of core antigen before treatment was significantly (P < .002) lower in patients with sustained response than in nonresponders. All responders had a sustained loss of core antigen, whereas all nonresponders remained core antigen positive. The concentrations of HCV core antigen and HCV RNA correlated significantly (n = 48, r = .627, P < .001). In conclusion, the HCV core antigen assay is useful for the diagnosis of acute and chronic hepatitis C, and for predicting and monitoring the effect of interferon alfa treatment. C1 Shinshu Univ, Sch Med, Dept Internal Med 2, Matsumoto, Nagano 3908621, Japan. Adv Life Sci Inst, Saitama, Japan. Shinshu Univ, Grad Sch Med, Inst Organ Transplants, Matsumoto, Nagano, Japan. Shinshu Univ, Grad Sch Med, Inst Recontruct Med, Matsumoto, Nagano, Japan. Shinshu Univ, Grad Sch Med, Inst Tissue Engn, Matsumoto, Nagano, Japan. NIH, Ctr Clin, Dept Transfus Med, Bethesda, MD 20892 USA. RP Tanaka, E (reprint author), Shinshu Univ, Sch Med, Dept Internal Med 2, 3-1-1 Asahi, Matsumoto, Nagano 3908621, Japan. NR 31 TC 113 Z9 116 U1 0 U2 3 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 2000 VL 32 IS 2 BP 388 EP 393 DI 10.1053/jhep.2000.9112 PG 6 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 350FR UT WOS:000089090700028 PM 10915747 ER PT J AU Lippincott-Schwartz, J Zaal, KJM AF Lippincott-Schwartz, J Zaal, KJM TI Cell cycle maintenance and biogenesis of the Golgi complex SO HISTOCHEMISTRY AND CELL BIOLOGY LA English DT Article; Proceedings Paper CT 42nd Symposium of the Society-for-Histochemistry CY SEP 23, 2000 CL LES DIABLERETS, SWITZERLAND SP Soc Histochem DE Golgi complex; mitosis; Golgi/ER recycling ID DOCKING PROTEIN P115; ENDOPLASMIC-RETICULUM; BREFELDIN-A; LIVING CELLS; RETROGRADE TRANSPORT; INTRACELLULAR-TRANSPORT; GUANINE-NUCLEOTIDE; RECYCLING PATHWAY; MATRIX PROTEIN; KDEL RECEPTOR AB How organelle identity is established and maintained, and how organelles divide and partition between daughter cells, are central questions of organelle biology. For the membrane-bound organelles of the secretory and endocytic pathways [including the endoplasmic reticulum (ER), Golgi complete, lysosomes, and endosomes], answering these questions has proved difficult because these organelles undergo continuous exchange of material. As a result, many "resident" proteins are not localized to a single site, organelle boundaries overlap, and when interorganellar membrane flow is interrupted, organelle structure is altered. The existence and identity of these organelles, therefore, appears to be a product of the dynamic processes of membrane trafficking and sorting. This is particularly true for the Golgi complex, which resides and functions at the crossroads of the secretory pathway. The Golgi receives newly synthesized proteins from the ER, covalently modifies them, and then distributes them to various final destinations within the cell. In addition, the Golgi recycles selected components back to the ER. These activities result from the Golgi's distinctive membranes, which are organized as polarized stacks (cis to trans) of flattened cisternae surrounded by tubules and vesicles. Golgi membranes are highly dynamic despite their characteristic organization and morphology, undergoing rapid disassembly and reassembly during mitosis and in response to perturbations in membrane trafficking pathways. How Golgi membranes fragment and disperse under these conditions is only beginning to be clarified, but is central to understanding the mechanism(s) underlying Golgi identity and biogenesis. Recent work, discussed in this review, suggests that membrane recycling pathways operating between the Golgi and ER play an indispensable role in Golgi maintenance and biogenesis, with the Golgi dispersing and reforming through the intermediary of the ER both in mitosis and in interphase when membrane cycling pathways are disrupted. C1 NICHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Lippincott-Schwartz, J (reprint author), NICHD, Cell Biol & Metab Branch, NIH, 18 Lib Dr,Bldg 18, Bethesda, MD 20892 USA. NR 66 TC 38 Z9 39 U1 0 U2 5 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0948-6143 J9 HISTOCHEM CELL BIOL JI Histochem. Cell Biol. PD AUG PY 2000 VL 114 IS 2 BP 93 EP 103 PG 11 WC Cell Biology; Microscopy SC Cell Biology; Microscopy GA 352JN UT WOS:000089211900002 PM 11052258 ER PT J AU Yao, RT Seidel, J Johnson, CA Daube-Witherspoon, ME Green, MV Carson, RE AF Yao, RT Seidel, J Johnson, CA Daube-Witherspoon, ME Green, MV Carson, RE TI Performance characteristics of the 3-D OSEM algorithm in the reconstruction of small animal PET images SO IEEE TRANSACTIONS ON MEDICAL IMAGING LA English DT Article DE ordered-subsets expectation-maximization; (OSEM) algorithm; reconstruction; small animal PET imaging; three-dimensional (3-D) volume imaging ID HIGH-RESOLUTION; EMISSION; SCANNER; QUANTITATION; TOMOGRAPHY AB Rat brain images acquired with a small animal positron emission tomography (PET) camera and reconstructed with the three-dimensional (3-D) ordered-subsets expectation-maximization (OSEM) algorithm with resolution recovery have better quality when the brain is imaged by itself than when inside the head with surrounding background activity. The purpose of this study was to characterize the dependence of this effect on the level of background activity, attenuation, and scatter, Monte Carlo simulations of the imaging system were performed. The coefficient of variation from replicate images, full-width at half-maximum (FWHM) from point sources and image profile fitting, and image contrast and uniformity were used to evaluate algorithm performance. A rat head with the typical levels of five and ten times the brain activity in the surrounding background requires additional iterations to achieve the same resolution as the brain-only case at a cost of 24% and 64% additional noise, respectively, For the same phantoms, object scatter reduced contrast by 3%-5%. However, attenuation degraded resolution by 0.2 mm and was responsible for up to 12% nonuniformity in the brain images suggesting that attenuation correction is useful, Given the effects of emission and attenuation distribution on both resolution and noise, simulations or phantom studies should be used for each imaging situation to select the appropriate number of OSEM iterations to achieve the desired resolution-noise levels. C1 NIH, PET Dept, Bethesda, MD 20892 USA. NIH, Dept Nucl Med, Bethesda, MD 20892 USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. Univ Penn, Dept Radiol, Philadelphia, PA 19104 USA. RP Yao, RT (reprint author), NIH, PET Dept, Bldg 10, Bethesda, MD 20892 USA. RI Carson, Richard/H-3250-2011 OI Carson, Richard/0000-0002-9338-7966 NR 26 TC 44 Z9 44 U1 0 U2 3 PU IEEE-INST ELECTRICAL ELECTRONICS ENGINEERS INC PI NEW YORK PA 345 E 47TH ST, NEW YORK, NY 10017-2394 USA SN 0278-0062 J9 IEEE T MED IMAGING JI IEEE Trans. Med. Imaging PD AUG PY 2000 VL 19 IS 8 BP 798 EP 804 DI 10.1109/42.876305 PG 7 WC Computer Science, Interdisciplinary Applications; Engineering, Biomedical; Engineering, Electrical & Electronic; Imaging Science & Photographic Technology; Radiology, Nuclear Medicine & Medical Imaging SC Computer Science; Engineering; Imaging Science & Photographic Technology; Radiology, Nuclear Medicine & Medical Imaging GA 367CY UT WOS:000090045200003 PM 11055803 ER PT J AU Seidel, J Vaquero, JJ Barbosa, F Lee, IJ Cuevas, C Green, MV AF Seidel, J Vaquero, JJ Barbosa, F Lee, IJ Cuevas, C Green, MV TI Scintillator identification and performance characteristics of LSO and GSO PSPMT detector modules combined through common X and Y resistive dividers SO IEEE TRANSACTIONS ON NUCLEAR SCIENCE LA English DT Article ID DEPTH AB Combining signal channels from detector arrays can reduce complexity and minimize cost but, potentially, at the expense of other performance parameters. We evaluated a method that reduces the number of signals by combining the anode outputs of three position-sensitive photomultiplier tubes (PSPMTs) through a common Ii resistive charge divider and three individual Y resistive charge dividers. Field flood images at 511 keV of two LSO modules combined with a single GSO module were compared to images obtained when the modules were illuminated separately. At moderate count rates only a small reduction in position detection accuracy was observed in the combined tubes. Event mis-positioning was minimal for total count rates < 300,000 cps. At higher rates, pulse pileup degraded accuracy. Delayed charge integration, a method for identifying scintillators by differences in their light decay times, allowed the LSO and GSO arrays to be distinguished from one another and also reduced the effect of pulse pileup. Thus, combining PSPMTs anodes through common X and common Y resistive dividers may be useful in reducing signal number from PSPMT detector modules while maintaining good event localization and scintillator identification accuracy at reasonable event rates. C1 NIH, Bethesda, MD 20892 USA. Thomas Jefferson Natl Accelerator Facil, Newport News, VA 23606 USA. RP Seidel, J (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. RI Vaquero, Juan Jose/D-3033-2009 OI Vaquero, Juan Jose/0000-0001-9200-361X NR 7 TC 9 Z9 9 U1 0 U2 0 PU IEEE-INST ELECTRICAL ELECTRONICS ENGINEERS INC PI NEW YORK PA 345 E 47TH ST, NEW YORK, NY 10017-2394 USA SN 0018-9499 J9 IEEE T NUCL SCI JI IEEE Trans. Nucl. Sci. PD AUG PY 2000 VL 47 IS 4 BP 1640 EP 1645 DI 10.1109/23.873028 PN 2 PG 6 WC Engineering, Electrical & Electronic; Nuclear Science & Technology SC Engineering; Nuclear Science & Technology GA 358UW UT WOS:000089576400017 ER PT J AU Smith, MF AF Smith, MF TI The effect of contraction and twist on myocardial PET and SPECT image resolution: A mathematical phantom study SO IEEE TRANSACTIONS ON NUCLEAR SCIENCE LA English DT Article ID EMISSION COMPUTED-TOMOGRAPHY; CARDIAC SPECT; MOTION; HEART; PARAMETER; QUANTIFICATION; MODELS; MRI AB Motion of the heart blurs cardiac PET and SPECT images. A mathematical phantom was constructed that models the contractile and non-uniform twisting motions of the left ventricular myocardium as the heart beats. The phantom was used to estimate the effect of normal heart motion on ECT image resolution and quantitation in regions of interest (ROIs). The movement of mid-myocardial points was tracked, from which motion blurring was computed. Clinical PET resolution of 7 mm was degraded to 10-12 mm FWHM in the direction of motion and clinical SPECT resolution of 14 mm to 15-16 mm FWHM. Contractile motion was greater near the base and twisting motion was greater near the apex. The blur due to both motions was equivalent to 7-10 mm FWHM Gaussian smoothing in the direction of motion. Activity weight coefficients characterized how activity in source model ROIs contributed to activity in short and long axis ROIs. The effect of contraction was dependent on whether the ROIs were from short or long axis slices and on the ROI location. The activity weights for short axis slices showed the effect of twist in blurring activity to adjacent angular sectors. This effect was not seen for constant thickness longs axis slices because the angular rang of the slices was greater, particularly near the apex. The contrast of perfusion defects in short axis ROIs was decreased by cardiac twist. A mathematical heart phantom that models contractile and twisting motions is useful for investigating the effect of these motions on cardiac ECT studies. C1 NIH, Dept Nucl Med, Ctr Clin, Bethesda, MD 20892 USA. RP Smith, MF (reprint author), NIH, Dept Nucl Med, Ctr Clin, Bldg 10, Bethesda, MD 20892 USA. NR 25 TC 2 Z9 2 U1 0 U2 2 PU IEEE-INST ELECTRICAL ELECTRONICS ENGINEERS INC PI NEW YORK PA 345 E 47TH ST, NEW YORK, NY 10017-2394 USA SN 0018-9499 J9 IEEE T NUCL SCI JI IEEE Trans. Nucl. Sci. PD AUG PY 2000 VL 47 IS 4 BP 1646 EP 1654 DI 10.1109/23.873029 PN 2 PG 9 WC Engineering, Electrical & Electronic; Nuclear Science & Technology SC Engineering; Nuclear Science & Technology GA 358UW UT WOS:000089576400018 ER PT J AU Tamura, T Nagamura-Inoue, T Shmeltzer, Z Kuwata, T Ozato, K AF Tamura, T Nagamura-Inoue, T Shmeltzer, Z Kuwata, T Ozato, K TI ICSBP directs bipotential myeloid progenitor cells to differentiate into mature macrophages SO IMMUNITY LA English DT Article ID SEQUENCE-BINDING-PROTEIN; TRANSCRIPTION FACTOR; IFN-GAMMA; REGULATORY FACTORS; DEFICIENT MICE; NITRIC-OXIDE; EXPRESSION; ACTIVATION; INDUCTION; PU.1 AB During hematopoiesis, myeloid progenitor cells give rise to granulocytes and macrophages. To study the role for ICSBP, a hematopoietic cell-specific transcription factor in myeloid cell development, the gene was introduced into myeloid progenitor cells established from ICSBP-/- mice. ICSBP retrovirus-transduced cells differentiated into mature macrophages with phagocytic activity, which coincided with the induction of specific target DNA binding activity. Similar to macrophages in vivo, ICSBP-transduced cells were growth arrested, expressed many macrophage-specific genes, and responded to macrophage activation signals. Contrary to this, ICSBP transduction led to repression of granulocyte-specific genes and inhibited G-CSF-mediated granulocytic differentiation in these and other myeloid progenitor cells. Together, ICSBP has a key role in the myeloid cell lineage selection and macrophage maturation. C1 NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Ozato, K (reprint author), NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. NR 52 TC 188 Z9 190 U1 0 U2 9 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 1074-7613 J9 IMMUNITY JI Immunity PD AUG PY 2000 VL 13 IS 2 BP 155 EP 165 DI 10.1016/S1074-7613(00)00016-9 PG 11 WC Immunology SC Immunology GA 350ZT UT WOS:000089132100001 PM 10981959 ER PT J AU Shaffer, AL Yu, X He, YS Boldrick, J Chan, EP Staudt, LM AF Shaffer, AL Yu, X He, YS Boldrick, J Chan, EP Staudt, LM TI BCL-6 represses genes that function in lymphocyte differentiation, inflammation, and cell cycle control SO IMMUNITY LA English DT Article ID GERMINAL-CENTER FORMATION; DEPENDENT KINASE INHIBITOR; CENTER B-CELLS; TRANSCRIPTIONAL REPRESSION; MICE LACKING; EXPRESSION; PROTEIN; P27(KIP1); MIP-1-ALPHA; BLIMP-1 AB BCL-6, a transcriptional repressor frequently translocated in lymphomas, regulates germinal center B cell differentiation and inflammation. DNA microarray screening identified genes repressed by BCL-6, including many lymphocyte activation genes, suggesting that BCL-6 modulates B cell receptor signals. BCL-6 repression of two chemokine genes, MIP-1 alpha and IP-10, may also attenuate inflammatory responses. Blimp-1, another BCL-6 target, is important for plasmacytic differentiation. Since BCL-6 expression is silenced in plasma cells, repression of blimp-1 by BCL-6 may control plasmacytic differentiation. Indeed, inhibition of BCL-6 function initiated changes indicative of plasmacytic differentiation, including decreased expression of c-Myc and increased expression of the cell cycle inhibitor p27kip1. These data suggest that malignant transformation by BCL-6 involves inhibition of differentiation and enhanced proliferation. C1 NCI, Metab Branch, Div Clin Sci, Bethesda, MD 20892 USA. RP Staudt, LM (reprint author), NCI, Metab Branch, Div Clin Sci, Bethesda, MD 20892 USA. NR 52 TC 547 Z9 557 U1 0 U2 8 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 1074-7613 J9 IMMUNITY JI Immunity PD AUG PY 2000 VL 13 IS 2 BP 199 EP 212 DI 10.1016/S1074-7613(00)00020-0 PG 14 WC Immunology SC Immunology GA 350ZT UT WOS:000089132100005 PM 10981963 ER PT J AU Notarangelo, LD Giliani, S Mella, P Schumacher, RF Mazza, C Savoldi, G Rodriguez-Perez, C Badolato, R Mazzolari, E Porta, F Candotti, F Ugazio, AG AF Notarangelo, LD Giliani, S Mella, P Schumacher, RF Mazza, C Savoldi, G Rodriguez-Perez, C Badolato, R Mazzolari, E Porta, F Candotti, F Ugazio, AG TI Combined immunodeficiencies due to defects in signal transduction: Defects of the gamma(c)-JAK3 signaling pathway as a model SO IMMUNOBIOLOGY LA English DT Article; Proceedings Paper CT XXVIIth Annual Meeting of the Clinical Immunology Working Group of the German-Society-for-Immunology CY NOV, 1999 CL FRANKFURT, GERMANY SP German Soc Immunol, Clin Immunol Working Grp ID RECEPTOR-GAMMA-CHAIN; HEMATOPOIETIC PROGENITOR CELLS; IN-UTERO TRANSPLANTATION; PROTEIN-TYROSINE KINASES; MICE LACKING JAK3; C GENE-TRANSFER; INTERLEUKIN-2 RECEPTOR; T-CELLS; IL-2 RECEPTOR; LYMPHOID DEVELOPMENT AB Combined immune deficiencies comprise a spectrum of genetic disorders characterized by developmental or functional defects of both T and B lymphocytes. Recent progress in cell biology and molecular genetics has unraveled the pathophysiology of most of these defects. In particular, the most common form of severe combined immune deficiency in humans, with lack of circulating T cells, a normal or increased number of B lymphocytes, and an X-linked pattern of inheritance (SCIDX1) has been shown to be due to defects of the IL2RG gene, encoding for the common gamma chain (gamma(c)), shared by several cytokine receptors. Furthermore, defects of the JAK3 gene, encoding for an intracellular tyrosine kinase required for signal transduction through gamma(c)-containing cytokine receptors, have been identified in patients with autosomal recessive T-B+ SCID. Characterization of the functional properties of cytokines that signal through the gamma(c)-JAK3 signaling pathway has been favored by the detailed analysis of SCID patients. Specifically, the key role of IL-7 in promoting T cell development has been substantiated by the identification of rare patients with T-B+ SCID who have a defect in the alpha subunit of the IL-7 receptor (IL7R alpha). The heterogeneity of genetic defects along the same signaling pathway that may lead to combined immune deficiency is paralleled by the heterogeneity of immunological phenotypes that may associate with defects in the same gene, thus creating a need for detailed immunological and molecular investigations in order to dissect the spectrum of combined immune deficiencies in humans. C1 Univ Brescia, Ist Med Mol Angelo Nocivelli, Pediat Clin, I-25121 Brescia, Italy. NHGRI, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA. RP Notarangelo, LD (reprint author), Univ Brescia, Pediat Clin, Spedali Civili, I-25123 Brescia, Italy. RI Badolato, Raffaele/A-8081-2010; Notarangelo, Luigi/F-9718-2016 OI Badolato, Raffaele/0000-0001-7375-5410; Notarangelo, Luigi/0000-0002-8335-0262 NR 78 TC 14 Z9 14 U1 0 U2 4 PU GUSTAV FISCHER VERLAG PI JENA PA VILLENGANG 2, D-07745 JENA, GERMANY SN 0171-2985 J9 IMMUNOBIOLOGY JI Immunobiology PD AUG PY 2000 VL 202 IS 2 BP 106 EP 119 PG 14 WC Immunology SC Immunology GA 345KE UT WOS:000088814500002 PM 10993286 ER PT J AU Garcia-Gonzalez, MA Crusius, JBA Strunk, MHP Bouma, G Perez-Centeno, CM Pals, G Meuwissen, SGM Pena, AS AF Garcia-Gonzalez, MA Crusius, JBA Strunk, MHP Bouma, G Perez-Centeno, CM Pals, G Meuwissen, SGM Pena, AS TI TGFB1 gene polymorphisms and inflammatory bowel disease SO IMMUNOGENETICS LA English DT Article DE inflammatory bowel disease; Crohn's disease; ulcerative colitis; transforming growth factor; polymorphism ID GROWTH-FACTOR-BETA; TRANSFORMING GROWTH-FACTOR-BETA-1 GENE; BLOOD-PRESSURE; CROHNS-DISEASE; ASSOCIATION; EXPRESSION; FIBROSIS C1 Univ Hosp Vrije Univ, Dept Gastroenterol, NL-1081 BT Amsterdam, Netherlands. Univ Hosp Vrije Univ, Lab Gastrointestinal Immunogenet, NL-1081 BT Amsterdam, Netherlands. Univ Hosp Vrije Univ, Dept Clin Genet, NL-1081 BT Amsterdam, Netherlands. NIAID, Mucosal Immun Sect, NIH, Bethesda, MD 20892 USA. RP Pena, AS (reprint author), Univ Hosp Vrije Univ, Dept Gastroenterol, Van Boechorststr 7, NL-1081 BT Amsterdam, Netherlands. RI Pals, Gerard/A-5198-2011; bouma, gerd/E-2520-2013; Strunk, Mark/K-3597-2014 OI Strunk, Mark/0000-0002-8619-3979 NR 22 TC 18 Z9 18 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0093-7711 J9 IMMUNOGENETICS JI Immunogenetics PD AUG PY 2000 VL 51 IS 10 BP 869 EP 872 DI 10.1007/s002510000211 PG 4 WC Genetics & Heredity; Immunology SC Genetics & Heredity; Immunology GA 346BY UT WOS:000088851700014 PM 10970103 ER PT J AU Van Kirk, LS Hayes, SF Heinzen, RA AF Van Kirk, LS Hayes, SF Heinzen, RA TI Ultrastructure of Rickettsia rickettsii actin tails and localization of cytoskeletal proteins SO INFECTION AND IMMUNITY LA English DT Article ID CHICKEN-EMBRYO FIBROBLASTS; PROLINE-RICH REGION; LISTERIA-MONOCYTOGENES; SHIGELLA-FLEXNERI; SPOTTED-FEVER; VERO CELLS; MICROFILAMENT PROTEINS; INTERCELLULAR SPREAD; FILAMENT NUCLEATION; FOCAL ADHESION AB Actin-based motility (ABM) is a mechanism for intercellular spread that is utilized by vaccinia virus and the invasive bacteria within the genera Rickettsia, Listeria, and Shigella. Within the Rickettsia, ABM is confined to members of the spotted fever group (SFG), such as Rickettsia rickettsii, the agent of Rocky Mountain spotted fever. Infection by each agent induces the polymerization of host cell actin to form the typical F (filamentous)actin comet tail. Assembly of the actin tail propels the pathogen through the host cytosol and into cell membrane protrusions that can be engulfed by neighboring cells, initiating a new infectious cycle. Little is known about the structure and morphogenesis of the Rickettsia rickettsii actin tail relative to Shigella and Listeria actin tails. In this study we examined the ultrastructure of the rickettsial actin tail by confocal, scanning electron, and transmission electron microscopy. Confocal microscopy of rhodamine phalloidin-stained infected Vero cells revealed the typhus group rickettsiae, Rickettsia prowazekii and Rickettsia typhi, to have no actin tails and short (similar to 1- to 3-mu m) straight or hooked actin tails, respectively. The SFG rickettsia, R. rickettsii, displayed long actin tails (>10 mu m) that were frequently comprised of multiple, distinct actin bundles, wrapping around each other in a helical fashion. Transmission electron microscopy, in conjunction with myosin S1 subfragment decoration, revealed that the individual actin filaments of R. rickettsii tails are >1 mu m long, arranged roughly parallel to one another, and oriented with the fast-growing barbed end towards the rickettsial pole. Scanning electron microscopy of intracellular rickettsiae demonstrated R. rickettsii to have polar associations of cytoskeletal material and R. prowazekii to be devoid of cytoskeletal interactions. By indirect immunofluorescence, both R. rickettsii and Listeria monocytogenes actin tails were shown to contain the cytoskeletal proteins vasodilator-stimulated phosphoprotein profilin, vinculin, and filamin. However, rickettsial tails lacked ezrin, paxillin, and tropomyosin, proteins that were associated with actin tails of cytosolic or protrusion-bound Listeria. The unique ultrastructural and compositional characteristics of the R. rickettsii actin tail suggest that rickettsial ABM is mechanistically different from previously described microbial ABM systems. C1 Univ Wyoming, Dept Mol Biol, Laramie, WY 82071 USA. NIAID, Rocky Mt Lab, Microscopy Branch, Hamilton, MT 59840 USA. RP Heinzen, RA (reprint author), Univ Wyoming, Dept Mol Biol, Laramie, WY 82071 USA. FU NIAID NIH HHS [AI-43502-01] NR 52 TC 64 Z9 68 U1 3 U2 9 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 2000 VL 68 IS 8 BP 4706 EP 4713 DI 10.1128/IAI.68.8.4706-4713.2000 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 337AY UT WOS:000088336200045 PM 10899876 ER PT J AU Du, LD Kolenbrander, PE AF Du, LD Kolenbrander, PE TI Identification of saliva-regulated genes of Streptococcus gordonii DL1 by differential display using random arbitrarily primed PCR SO INFECTION AND IMMUNITY LA English DT Article ID ANTIGEN I/II; ORAL STREPTOCOCCI; PORPHYROMONAS-GINGIVALIS; ENTEROCOCCUS-FAECALIS; NUCLEOTIDE-SEQUENCE; SURFACE-ANTIGEN; SANGUIS; BINDING; POLYPEPTIDES; AGGLUTININ AB Attachment of Streptococcus gordonii to the acquired pellicle of the tooth surface involves specific interactions between bacterial adhesins and adsorbed salivary components. To study saliva-regulated gene expression in S. gordonii, we used random arbitrarily primed PCR (RAP-PCR). Bacteria were incubated in either brain heart infusion medium or saliva. Total RNA from both conditions was purified and RAP fingerprinted and then PCR amplified with an arbitrary primer. The differentially displayed DNA fragments were cloned, sequenced, and analyzed using the BLAST search network service. Three DNA products were up-regulated. One was identified as that of the sspA and -B genes, which encode the salivary agglutinin glycoprotein-binding proteins SspA and SspB of S. gordonii; another had 79% identity with the Lactococcus lactis clpE gene, encoding a member of the Clp protease family; and the third product showed no significant homology to known genes. Five downregulated genes were identified which encode proteins involved in bacterial metabolism. We have shown, for the first time, direct induction of sspA and -B in S. gordonii by human saliva. C1 NIDCR, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Kolenbrander, PE (reprint author), NIDCR, Oral Infect & Immun Branch, NIH, Bldg 30,Room 310,30 COnvent Dr,MSC 4350, Bethesda, MD 20892 USA. NR 31 TC 44 Z9 47 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 2000 VL 68 IS 8 BP 4834 EP 4837 DI 10.1128/IAI.68.8.4834-4837.2000 PG 4 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 337AY UT WOS:000088336200070 PM 10899901 ER PT J AU Yu, ML Guo, YLL Hsu, CC Rogan, WJ AF Yu, ML Guo, YLL Hsu, CC Rogan, WJ TI Menstruation and reproduction in women with polychlorinated biphenyl (PCB) poisoning: long-term follow-up interviews of the women from the Taiwan Yucheng cohort SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE polychlorinated dibenzofurans; polychlorobiphenyl compounds; rice oil; infant mortality ID EXPOSURE; PCDFS; OIL AB Background In 1979 in Taiwan, about 2000 people were poisoned by cooking oil contaminated with heat-degraded polychlorinated biphenyls (PCBs). Victims developed chloracne and other symptoms; the illness is called Yucheng (oil disease). The chemicals persist in the victims' bodies and some are toxic to the female reproductive system in experimental animals. Methods In 1993-1994, we identified the living Yucheng women and neighbourhood controls and interviewed them about their reproductive experience. Of the 596 living Yucheng women aged 30-59 years, we found 368, and interviewed 356. We identified 329 controls, and interviewed 312. Results Of the Yucheng women, 16% reported abnormal menstrual bleeding compared to 8% (P < 0.05) of control women; 4.2% versus 1.7% reported a stillbirth since 1979 (P = 0.068). Other characteristics of the menses, fertility, frequency of intercourse, and age at menopause appeared unaffected. More of the Yucheng women reported that one of their offspring had died during childhood (10.2% versus 6.1%, P < 0.05), and that they had decided to limit childbearing because of health problems (7% versus 2%, P = 0.01). Conclusions These findings suggest that high level PCB/polychlorinated dibenzofuran (PCDF) exposure has some effect on female endocrine and reproductive function. C1 Natl Cheng Kung Univ, Coll Med, Dept Environm & Occupat Hlth, Tainan 70428, Taiwan. Natl Cheng Kung Univ, Coll Med, Dept Publ Hlth, Tainan, Taiwan. Tainan City Hosp, Dept Psychiat, Tainan, Taiwan. Natl Inst Environm Hlth Sci, Epidemiol Branch, Res Triangle Pk, NC USA. RP Guo, YLL (reprint author), Natl Cheng Kung Univ, Coll Med, Dept Environm & Occupat Hlth, 138 Sheng Li Rd, Tainan 70428, Taiwan. RI Rogan, Walter/I-6034-2012; OI Rogan, Walter/0000-0002-9302-0160; GUO, Yue Leon/0000-0002-8530-4809 NR 16 TC 72 Z9 73 U1 1 U2 7 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD AUG PY 2000 VL 29 IS 4 BP 672 EP 677 DI 10.1093/ije/29.4.672 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 349WW UT WOS:000089069400010 PM 10922344 ER PT J AU Lohman, TG Caballero, B Himes, JH Davis, CE Stewart, D Houtkooper, L Going, SB Hunsberger, S Weber, JL Reid, R Stephenson, L AF Lohman, TG Caballero, B Himes, JH Davis, CE Stewart, D Houtkooper, L Going, SB Hunsberger, S Weber, JL Reid, R Stephenson, L TI Estimation of body fat from anthropometry and bioelectrical impedance in Native American children SO INTERNATIONAL JOURNAL OF OBESITY LA English DT Article DE body fat; obesity; bioelectrical impedance; deuterium oxide; Native Americans ID CROSS-VALIDATION; WATER; MASS; FATNESS; ADOLESCENTS; EQUATIONS; YOUTH; INDEX; O-18 AB OBJECTIVE: Obesity, as measured by body mass index, is highly prevalent in Native American children, yet there are no valid equations to estimate total body fatness for this population. This study was designed to develop equations to estimate percentage body fat from anthropometry and bioelectrical impedance as a critical part of Pathways, a multisite study of primary prevention of obesity in Native American children. DESIGN: Percentage fat was estimated from deuterium oxide dilution in 98 Native American children (Pima/Maricopa, Tohono O'odham and White Mountain Apache tribes) between 8 and 11 y of age. The mean fat content (38.4% +/- 8.1%) was calculated assuming the water content of the fat-free body was 76%. Initial independent variables were height, weight, waist circumference, six skinfolds and whole-body resistance and reactance from bioelectrical impedance (BIA). RESULTS: Using all-possible-subsets regressions with the Mallows C (p) criterion, and with age and sex included in each regression model, waist circumference, calf and biceps skinfolds contributed least to the multiple regression analysis. The combination of weight, two skinfolds (any two out of the four best: triceps, suprailiac, subscapular and abdomen) and bioelectrical impedance variables provided excellent predictability. Equations without BIA variables yielded r(2) almost as high as those with BIA variables. The recommended equation predicts percentage fat with a root mean square error = 3.2% fat and an adjusted r(2) = 0.840. CONCLUSION: The combination of anthropometry and BIA variables can be used to estimate total body fat in field studies of Native American children. The derived equation yields considerably higher percentage fat values than other skinfold equations in children. C1 Univ Arizona, Tucson, AZ 85721 USA. Johns Hopkins Univ, Ctr Human Nutr, Baltimore, MD 21218 USA. Univ Minnesota, Minneapolis, MN 55455 USA. NHLBI, Bethesda, MD 20892 USA. Univ N Carolina, Chapel Hill, NC 27515 USA. RP Lohman, TG (reprint author), Univ Arizona, 114 Ina Gittings Bldg, Tucson, AZ 85721 USA. NR 31 TC 31 Z9 32 U1 1 U2 4 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0307-0565 J9 INT J OBESITY JI Int. J. Obes. PD AUG PY 2000 VL 24 IS 8 BP 982 EP 988 DI 10.1038/sj.ijo.0801318 PG 7 WC Endocrinology & Metabolism; Nutrition & Dietetics SC Endocrinology & Metabolism; Nutrition & Dietetics GA 341NT UT WOS:000088597000007 PM 10951536 ER PT J AU Turcato, E Bosello, O Di Francesco, V Harris, TB Zoico, E Bissoli, L Fracassi, E Zamboni, M AF Turcato, E Bosello, O Di Francesco, V Harris, TB Zoico, E Bissoli, L Fracassi, E Zamboni, M TI Waist circumference and abdominal sagittal diameter as surrogates of body fat distribution in the elderly: their relation with cardiovascular risk factors SO INTERNATIONAL JOURNAL OF OBESITY LA English DT Article DE body fat distribution; aging; waist; sagittal diameter; metabolic variables ID VISCERAL ADIPOSE-TISSUE; DIFFERENT ANTHROPOMETRIC MEASUREMENTS; COMPUTED-TOMOGRAPHY; WOMEN; MEN; LIPIDS; OBESE; AGE; ACCUMULATION; LIPOPROTEINS AB OBJECTIVE: To evaluate the relationship between supine sagittal abdominal diameter (SAD) and other indicators of body fat distribution with cardiovascular (CVD) risk factors in the elderly. SUBJECTS: One-hundred and forty-six women aged from 67 to 78 y with a body mass index (BMI) ranging from 18.7 to 50.6 kg/m(2) and 83 men aged between 67 and 78 y with BMI ranging from 19.8 to 37.1 kg/m(2). MEASUREMENT: Body fat distribution was assessed using anthropometric indicators: waist circumference, SAD, waist-to-hip ratio (WHR), waist-to-height ratio and SAD-to-thigh ratio. RESULTS: In women, there was a negative correlation between HDL-cholesterol and body weight, BMI, waist, SAD, WHR, waist-to-height ratio and SAD-to-thigh ratio. A significant association was found between triglycerides, basal glucose, 2 h glucose during oral glucose tolerance test (OGTT), systolic blood pressure (SBP), diastolic blood pressure (DBP) and anthropometric variables. In men a negative correlation was found between HDL-cholesterol and all the anthropometric variables. A significant association was found between triglycerides, DBP and body weight, BMI, waist, SAD and waist-to-height ratio. In women, after adjusting for age and BMI, a significant correlation was observed between waist and HDL-cholesterol, triglycerides and basal glucose. This was also seen with SAD and SAD-to-thigh ratio and triglycerides, basal and 2 h glucose. In men, after adjusting for age and BMI a significant correlation was found between SAD and HDL-cholesterol and triglycerides. When adjustments were made for age and waist, the correlations between BMI and metabolic variables as well between BMI and SEP and DBP were no longer significant. CONCLUSIONS: Our study shows that indicators of body fat distribution are associated with CVD risk factors in the elderly independently of BMI. Our data also show that waist and SAD are the anthropometric indicators of fat distribution which are most closely related to CVD risk factors in old age. C1 Univ Verona, Inst Geriatr Med, I-37126 Verona, Italy. NIA, Off Geriatr Epidemiol, Epidemiol & Demog & Biometry Program, Bethesda, MD 20892 USA. RP Zamboni, M (reprint author), Univ Verona, Osped Maggiore, Inst Geriatr Med, Piazzale Stefani 1, I-37126 Verona, Italy. OI ZAMBONI, Mauro/0000-0001-6961-9483; ZOICO, Elena/0000-0002-5235-1545 NR 31 TC 98 Z9 103 U1 2 U2 7 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0307-0565 J9 INT J OBESITY JI Int. J. Obes. PD AUG PY 2000 VL 24 IS 8 BP 1005 EP 1010 DI 10.1038/sj.ijo.0801352 PG 6 WC Endocrinology & Metabolism; Nutrition & Dietetics SC Endocrinology & Metabolism; Nutrition & Dietetics GA 341NT UT WOS:000088597000010 PM 10951539 ER PT J AU Zhong, XS Thornton, K Reed, E AF Zhong, XS Thornton, K Reed, E TI Computer based analyses of the 5 '-flanking regions of selected genes involved in the nucleotide excision repair complex SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE nucleotide excision repair; ovarian cancer; testicular cancer; brain rumors ID MESSENGER-RNA EXPRESSION; INTERSTRAND CROSS-LINKS; OVARIAN-CARCINOMA CELLS; YEAST DNA-REPAIR; COMPLEMENTATION GROUP-1; TUMOR-CELLS; CISPLATIN; ERCC-1; PROTEIN; LINES AB We have previously observed that the mRNA of selected genes involved in nucleotide excision repair appear to be coordinately expressed in human tissues from patients with ovarian cancer, testicular cancer, malignant blain tumors, and other malignancies. Such genes include ERCC1, XPA, XPB, XPD, XPF, and XPG. Coordinate mRNA expression appeals to be most impressive in non-malignant tissues. We therefore began to explore possible reasons why such coordinate expression should occur. DNA sequences for the above noted genes were obtained from GeneBank(TM). Two different software programs were applied to the DNA sequence, to the area 5' to the start of exon I of each gene. Analyses were performed by computer. The length of the 5' area assessed, was based on previous reports that determined what portion of the genomic sequence comprised the 5' UTR of the promoter of the respective gene. Based on this approach, potential DNA binding sites for no less than three dozen proteins, were identified in the 5'-flanking legion of each of the NER genes studied. For each gene, potential binding sites for activator proteins and for repressor proteins were identified. The 5'-flanking, regions for each gene noted above, had binding sites in common for 14 proteins with transcription modulatory activity. Eleven of these proteins are known for activator activity; two are reported to have repressor activity, and one has both repressor and activator function. These data suggest a possible molecular basis for the previously observed coordinate mRNA expression of selected NER genes in human tissue specimens. C1 NCI, Med Ovarian Canc Sect, Med Branch, DCS, Bethesda, MD 20892 USA. RP Reed, E (reprint author), NCI, Med Ovarian Canc Sect, Med Branch, DCS, Bldg 10,Room 12N226, Bethesda, MD 20892 USA. NR 32 TC 11 Z9 11 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 2000 VL 17 IS 2 BP 375 EP 380 PG 6 WC Oncology SC Oncology GA 334WM UT WOS:000088208700023 PM 10891549 ER PT J AU Rodriguez, IR Moreira, EF Bok, D Kantorow, M AF Rodriguez, IR Moreira, EF Bok, D Kantorow, M TI Osteonectin/SPARC secreted by RPE and localized to the outer plexiform layer of the monkey retina SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID PROTEIN SPARC; GROWTH-FACTOR; BINDING; GLYCOPROTEIN; EXPRESSION; BM-40; GENE; COLLAGEN; DOMAINS; TISSUES AB PURPOSE. Osteonectin/SPARC is a secreted protein that has been implicated in ocular disease. Deletion of osteonectin/SPARC causes age-onset cataract in mice and the cataractous human lens has increased expression of osteonectin/SPARC. In this study, the expression and localization of osteonectin/SPARC in the monkey retina were determined as was secretion by cultured human retinal pigment epithelial (RPE) cells. METHODS. Adult Rhesus monkey eyes (Macaca mulatta) were dissected, and 5-mm macula and peripheral retina punches were obtained. Supernatants were collected from cultured human RPE cells. Subcellular fractionation of whole monkey retina was also performed. Osteonectin/SPARC expression and/or secretion was monitored by Northern and Western blot analyses, and localization was determined by immunocytochemistry. RESULTS. Outside of the retina osteonectin/SPARC mRNA is broadly expressed in many human tissues. Northern blot analysis shows that in the retina osteonectin/SPARC is expressed almost exclusively by the macular RPE/choroid. Western blot analysis revealed osteonectin/SPARC in both the macula and the peripheral neural retina but only in trace amounts in the RPE/choroid. In subcellular fractions of the whole retina, osteonectin/SPARC was detected, mainly in the soluble fraction but also in the membrane and nuclear fractions. Immunohistochemical analysis localized osteonectin/SPARC specifically to the outer plexiform layer. Western blot analysis of conditioned medium from human RPE cells cultured on porous substrates indicated that osteonectin/SPARC is secreted in large amounts from both the apical and basal sides of the RPE. CONCLUSIONS. Collectively these data provide evidence that osteonectin/SPARC is synthesized in the macular RPE, secreted, and subsequently transported to the outer plexiform layer. The expression pattern of osteonectin/SPARC in the subcellular retinal fractions is consistent with a soluble protein that is transported and internalized. C1 NEI, NIH, LRCMB, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Jules Stein Eye Inst, Los Angeles, CA 90024 USA. Univ Calif Los Angeles, Brain Res Inst, Los Angeles, CA 90024 USA. Univ Calif Los Angeles, Dept Neurobiol, Los Angeles, CA USA. W Virginia Univ, Dept Biol, Morgantown, WV 26506 USA. RP Rodriguez, IR (reprint author), NEI, NIH, LRCMB, 6 Ctr Dr MSC 2740,Bldg 6,Room 304, Bethesda, MD 20892 USA. FU NEI NIH HHS [1R01EY13022, EY00331, P30 EY000331, R01 EY000444, R01 EY013022, R01 EY013022-01, Z01 EY000444] NR 28 TC 8 Z9 8 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 2000 VL 41 IS 9 BP 2438 EP 2444 PG 7 WC Ophthalmology SC Ophthalmology GA 340TT UT WOS:000088551600006 PM 10937551 ER PT J AU Furuya, K Kawanaka, M Sato, N Honma, H Tamura, M AF Furuya, K Kawanaka, M Sato, N Honma, H Tamura, M TI Has Echinococcus granulosus settled in Hokkaido? SO JAPANESE JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID ALVEOLAR ECHINOCOCCOSIS C1 Hokkaido Inst Publ Hlth, Kita Ku, Sapporo, Hokkaido 0600819, Japan. NIAID, Dept Parasitol, Shinjuku Ku, Tokyo 1628640, Japan. Hokkaido Univ, Sch Med, Kita Ku, Sapporo, Hokkaido 0608648, Japan. RP Furuya, K (reprint author), Hokkaido Inst Publ Hlth, Kita Ku, Kita-19,Nishi-12, Sapporo, Hokkaido 0600819, Japan. EM furuya@iph.pref.hokkaido.jp NR 7 TC 1 Z9 1 U1 0 U2 0 PU NATL INST INFECTIOUS DISEASES PI TOKYO PA JPN J INFECT DIS ED OFF NATL INST INFECTIOUS DISEASES TOYAMA 1-23-1, SHINJUKU-KU, TOKYO, 162-8640, JAPAN SN 1344-6304 EI 1884-2836 J9 JPN J INFECT DIS JI Jpn. J. Infect. Dis. PD AUG PY 2000 VL 53 IS 4 BP 176 EP 177 PG 2 WC Infectious Diseases SC Infectious Diseases GA 370RK UT WOS:000165135900011 PM 11056566 ER PT J AU Levenston, GK Patrick, CJ Bradley, MM Lang, PJ AF Levenston, GK Patrick, CJ Bradley, MM Lang, PJ TI The psychopath as observer: Emotion and attention in picture processing SO JOURNAL OF ABNORMAL PSYCHOLOGY LA English DT Article ID FEAR-POTENTIATED STARTLE; NONPSYCHOPATHIC OFFENDERS; CRIMINAL PSYCHOPATH; CONSTRUCT-VALIDITY; VISUAL-ATTENTION; ACTIVATION; STIMULUS; AROUSAL; REFLEX; PROBE AB This study extended prior work showing abnormal affect-startle modulation in psychopaths. Male prisoners viewed specific categories of pleasant (erotic or thrilling) and unpleasant (victim or direct threat) slide pictures, along with neutral pictures. Acoustic startle probes were presented early (300 and 800 ms) and late (1,800, 3,000, and 4,500 ms) in the viewing interval. At later times, nonpsychopaths showed moderate and strong reflex potentiation for victim and threat scenes, respectively. For psychopaths, startle was inhibited during victim scenes and only weakly potentiated during threat. Psychopaths also showed more reliable blink inhibition across pleasant contents than nonpsychopaths and greater heart rate orienting to affective pictures overall. These results indicate a heightened aversion threshold in psychopaths. In addition, deficient reflex modulation at early times suggested a weakness in initial stimulus evaluation among psychopaths. C1 Univ Minnesota, Dept Psychol, Minneapolis, MN 55455 USA. Florida State Univ, Dept Psychol, Tallahassee, FL 32306 USA. Univ Florida, NIMH, Ctr Study Emot & Attent, Gainesville, FL 32611 USA. Univ Florida, Dept Clin & Hlth Psychol, Gainesville, FL 32611 USA. RP Patrick, CJ (reprint author), Univ Minnesota, Dept Psychol, Minneapolis, MN 55455 USA. RI Frank, David/E-8213-2012 FU NIMH NIH HHS [MH48657, MH52384] NR 86 TC 210 Z9 211 U1 6 U2 64 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0021-843X J9 J ABNORM PSYCHOL JI J. Abnorm. Psychol. PD AUG PY 2000 VL 109 IS 3 BP 373 EP 385 DI 10.1037//0021-843X.109.3.373 PG 13 WC Psychology, Clinical; Psychology, Multidisciplinary SC Psychology GA 355WP UT WOS:000089411800003 PM 11016107 ER PT J AU Lahey, BB Schwab-Stone, M Goodman, SH Waldman, ID Canino, G Rathouz, PJ Miller, TL Dennis, KD Bird, H Jensen, PS AF Lahey, BB Schwab-Stone, M Goodman, SH Waldman, ID Canino, G Rathouz, PJ Miller, TL Dennis, KD Bird, H Jensen, PS TI Age and gender differences in oppositional behavior and conduct problems: A cross-sectional household study of middle childhood and adolescence SO JOURNAL OF ABNORMAL PSYCHOLOGY LA English DT Article ID DSM-III-R; ANTISOCIAL-BEHAVIOR; DEFIANT DISORDER; CRITERION VALIDITY; MENTAL-HEALTH; PSYCHOPATHOLOGY; PREVALENCE; CHILDREN; SAMPLE; GIRLS AB Behavior problems among youths cannot be understood without explaining their age and gender differences, bur age and gender differences cannot be explained until they have been accurately described. In a household survey of 1,285 youths aged 9 to 17 years, there were no gender differences in oppositional behavior. but aggression, property offenses, and status offenses were more common among boys. Levels of oppositional behavior were greater at younger ages, aggression peaked near the middle of this age range, and property and status of tenses were more prevalent at older ages. These findings are generally consistent with developmental models of conduct problems bur are inconsistent with a recent model of gender differences and raise questions about the external validity of current taxonomies. C1 Univ Chicago, Dept Psychiat, Chicago, IL 60637 USA. Yale Univ, Ctr Child Study, New Haven, CT 06520 USA. Emory Univ, Dept Psychol, Atlanta, GA 30322 USA. Univ Puerto Rico, Behav Sci Res Inst, San Juan, PR 00936 USA. Univ Chicago, Sch Med, Chicago, IL 60637 USA. Columbia Univ, Dept Psychiat, New York, NY 10027 USA. NIMH, Bethesda, MD 20892 USA. RP Lahey, BB (reprint author), Univ Chicago, Dept Psychiat, MC 3077,5841 S Maryland Ave, Chicago, IL 60637 USA. OI Jensen, Peter/0000-0003-2387-0650 FU NIMH NIH HHS [U01 MH46718, U01 MH46717, U01 MH46725] NR 52 TC 151 Z9 154 U1 7 U2 26 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0021-843X J9 J ABNORM PSYCHOL JI J. Abnorm. Psychol. PD AUG PY 2000 VL 109 IS 3 BP 488 EP 503 DI 10.1037//0021-843X.109.3.488 PG 16 WC Psychology, Clinical; Psychology, Multidisciplinary SC Psychology GA 355WP UT WOS:000089411800014 PM 11016118 ER PT J AU Gold, JM Rehkemper, G Binks, SW Carpenter, CJ Fleming, K Goldberg, TE Weinberger, DR AF Gold, JM Rehkemper, G Binks, SW Carpenter, CJ Fleming, K Goldberg, TE Weinberger, DR TI Learning and forgetting in schizophrenia SO JOURNAL OF ABNORMAL PSYCHOLOGY LA English DT Article ID TEMPORAL-LOBE; NEUROPSYCHOLOGICAL DEFICITS; MEMORY; IMPAIRMENT; IMMEDIATE; AMNESIA; RATES AB Recent studies of patients with schizophrenia have consistently demonstrated marked deficits on measures of initial learning. However, contradictory results have been reported concerning retention and forgetting. The present study examined the level of initial and delayed recall of stories and visual figures in a group of 76 patients with schizophrenia and 51 normal controls. schizophrenia patients demonstrated marked impairments in initial and delayed recall as well as significantly worse percentage retention scores. However. schizophrenia patients and healthy controls individually matched on level of initial recall bad nearly identical delayed recall performance. The results suggest a primary deficit in the initial acquisition of information rather than an accelerated rate of forgetting in schizophrenia. C1 NIMH, St Elizabeths Hosp, Ctr Neurosci, Clin Res Serv Branch,Div Intramural Res Programs, Washington, DC 20032 USA. RP Gold, JM (reprint author), Maryland Psychiat Res Ctr, POB 21247, Baltimore, MD 21228 USA. NR 36 TC 44 Z9 44 U1 0 U2 0 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0021-843X J9 J ABNORM PSYCHOL JI J. Abnorm. Psychol. PD AUG PY 2000 VL 109 IS 3 BP 534 EP 538 DI 10.1037//0021-843X.109.3.534 PG 5 WC Psychology, Clinical; Psychology, Multidisciplinary SC Psychology GA 355WP UT WOS:000089411800019 PM 11016123 ER PT J AU Johnson, JD Ryan, MJ Toft, JD Graves, SW Hejtmancik, MR Cunningham, ML Herbert, R Abdo, KM AF Johnson, JD Ryan, MJ Toft, JD Graves, SW Hejtmancik, MR Cunningham, ML Herbert, R Abdo, KM TI Two-year toxicity and carcinogenicity study of methyleugenol in F344/N rats and B6C3F(1) mice SO JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY LA English DT Article DE methyleugenol; toxicity; carcinogenicity; toxicokinetics; rats; mice ID NATURALLY-OCCURRING ALKENYLBENZENES; UNSCHEDULED DNA-SYNTHESIS; POST-LABELING ANALYSIS; SAFROLE; MOUSE; ESTRAGOLE; TESTS; HEPATOBLASTOMA; MUTAGENICITY; DERIVATIVES AB Methyleugenol (MEG) was tested for toxicity/carcinogenicity in a 2-yr carcinogenesis bioassay because of its widespread use in a variety of foods, beverages, and cosmetics as well as its structural resemblance to the known carcinogen safrole. F344/N rats and B6C3F(1) mice (50 animals/sex/dose group) were given MEG suspended in 0.5% methylcellulose by gavage at doses of 37, 75, or 150 mg/kg/day for 2 yr. Control groups (60 rats/sex and 50 mice/sex) received only the vehicle. A stop-exposure group of 60 rats/sex received 300 mg/kg/day by gavage for 53 weeks followed by the vehicle only for the remaining 52 weeks of the study. A special study group (10 animals/sex/species/dose group) were used for toxicokinetic studies. All male rats given 150 and 300 mg/kg/day died before the end of the study; survival of female rats given 150 mg/kg/day and all treated female mice was decreased. Mean body weights of treated male and female rats and mice were decreased when compared to control. Area under the curve results indicated that greater than dose proportional increases in plasma MEG occurred for male 150 and 300 mg/kg/day group rats (6 and 12 month) and male 150 mg/kg/day mice (12 month). Target organs included the liver, glandular stomach, forestomach (female rats) and kidney, mammary gland, and subcutaneous tissue (male rats). Liver neoplasms occurred in all dose groups of rats and mice and included hepatoadenoma, hepatocarcinoma, hepatocholangioma (rats only), hepatocholangiocarcinoma, and hepatoblastoma (mice only). Nonneoplastic liver lesions included eosinophilic and mixed cell foci (rats only), hypertrophy, oval cell hyperplasia, cystic degeneration (rats only), and bile duct hyperplasia. Mice also exhibited necrosis, hematopoietic cell proliferation, and hemosiderin pigmentation. Glandular stomach lesions in rats and mice included benign and malignant neuroendocrine tumors, neuroendocrine cell hyperplasia, and atrophy and in mice included glandular ectasia/chronic active inflammation. In female rats, the forestomach showed a positive trend in the incidences of squamous cell papilloma or carcinoma (combined). Male rats also exhibited kidney (renal tubule hyperplasia, nephropathy, and adenomacarcinoma), mammary gland (fibroadenoma), and subcutaneous tissue (fibroma, fibrosarcoma) lesions. Male rats also exhibited malignant mesotheliomas and splenic fibrosis. These data demonstrate that MEG is a multisite, multispecies carcinogen. C1 Battelle Columbus Labs, Columbus, OH 43201 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Abdo, KM (reprint author), Battelle Columbus Labs, 505 King Ave, Columbus, OH 43201 USA. NR 63 TC 25 Z9 29 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0021-8561 J9 J AGR FOOD CHEM JI J. Agric. Food Chem. PD AUG PY 2000 VL 48 IS 8 BP 3620 EP 3632 DI 10.1021/jf000364a PG 13 WC Agriculture, Multidisciplinary; Chemistry, Applied; Food Science & Technology SC Agriculture; Chemistry; Food Science & Technology GA 347VU UT WOS:000088950500092 PM 10956160 ER PT J AU Muller, FMC Weig, M Peter, J Walsh, TJ AF Muller, FMC Weig, M Peter, J Walsh, TJ TI Azole cross-resistance to ketoconazole, fluconazole, itraconazole and voriconazole in clinical Candida albicans isolates from HIV-infected children with oropharyngeal candidosis SO JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY LA English DT Letter ID ORAL CANDIDOSIS C1 Univ Wurzburg, Univ Kinderklin, D-97080 Wurzburg, Germany. NCI, Immunocompromised Host Sect, Pediat Oncol Branch, NIH, Bethesda, MD 20892 USA. Univ Wurzburg, Inst Mol Infekt Biol, D-97080 Wurzburg, Germany. Univ Wurzburg, Inst Hyg & Mikrobiol, Wurzburg, Germany. RP Muller, FMC (reprint author), Univ Wurzburg, Univ Kinderklin, Josef Schneider Str 2, D-97080 Wurzburg, Germany. NR 6 TC 58 Z9 59 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-7453 J9 J ANTIMICROB CHEMOTH JI J. Antimicrob. Chemother. PD AUG PY 2000 VL 46 IS 2 BP 338 EP 341 DI 10.1093/jac/46.2.338 PG 4 WC Infectious Diseases; Microbiology; Pharmacology & Pharmacy SC Infectious Diseases; Microbiology; Pharmacology & Pharmacy GA 354BL UT WOS:000089309900038 PM 10933673 ER PT J AU Xue, HH Zhao, DM Suda, T Uchida, C Oda, T Chida, K Ichiyama, A Nakamura, H AF Xue, HH Zhao, DM Suda, T Uchida, C Oda, T Chida, K Ichiyama, A Nakamura, H TI Store depletion by caffeine/ryanodine activates capacitative Ca2+ entry in nonexcitable A549 cells SO JOURNAL OF BIOCHEMISTRY LA English DT Article DE capacitative Ca2+ entry; inositol 1,4,5-trisphosphate receptor; ryanodine receptor; transient receptor potential protein ID SMOOTH-MUSCLE CELLS; INOSITOL 1,4,5-TRISPHOSPHATE; CALCIUM STORES; RYANODINE RECEPTOR; PC12 CELLS; DIFFERENTIAL EXPRESSION; ENDOPLASMIC-RETICULUM; HIPPOCAMPAL-NEURONS; HUMAN-NEUTROPHILS; INSP(3) RECEPTOR AB Capacitative Ca2+ entry is essential for refilling intracellular Ca2+ stores and is thought to be regulated primarily by inositol 1, 4,5-trisphosphate (IP3)-sensitive stores in nonexcitable cells, In nonexcitable A549 cells, the application of caffeine or ryanodine induces Ca2+ release in the absence of extracellular Ca2+ similar to that induced by thapsigargin (Tg), and Ca2+ entry occurs upon the readdition of extracellular Ca2+. The channels thus activated are also permeable to Mn2+. The channels responsible for this effect appear to be activated by the depletion of caffeine/ryanodine-sensitive stores per se, as evidenced by the activation even in the absence of increased intracellular Ca2+ concentration. Tg pretreatment abrogates the response to caffeine/ryanodine, whereas Tg application subsequent to caffeine/ryanodine treatment induces further Ca2+ release. The response to caffeine/ryanodine is also abolished by initial ATP application, whereas ATP added subsequent to caffeine/ryanodine induces additional Ca2+ release. RT-PCR analyses showed the expression of a type 1 ryanodine receptor, two human homologues of transient receptor potential protein (hTrp1 and hTrp6), as well as all three types of the IP3 receptor. These results suggest that in A549 cells, (i) capacitative Ca2+ entry can also be regulated by caffeine/ryanodine-sensitive stores, and (ii) the RyR-gated stores interact functionally with those sensitive to IP3, probably via Ca2+-induced Ca2+ release. C1 Hamamatsu Univ Sch Med, Dept Biochem 1, Hamamatsu, Shizuoka 4313192, Japan. Hamamatsu Univ Sch Med, Dept Internal Med, Div 2, Hamamatsu, Shizuoka 4313192, Japan. RP Zhao, DM (reprint author), NHLBI, NIH, Lab Mol Immunol, 9000 Rockville Pike,Bldg 10,Room 7N246, Bethesda, MD 20892 USA. EM xueh@nhlbi.nih.gov NR 48 TC 14 Z9 14 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0021-924X J9 J BIOCHEM JI J. Biochem. PD AUG PY 2000 VL 128 IS 2 BP 329 EP 336 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 343NX UT WOS:000088709400023 PM 10920270 ER PT J AU Banerjee, RK Back, LH Back, MR Cho, YI AF Banerjee, RK Back, LH Back, MR Cho, YI TI Physiological flow simulation in residual human stenoses after coronary angioplasty SO JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME LA English DT Article ID RATE-PRESSURE-DROP; ARTERIAL STENOSES; PULSATILE FLOW; SHEAR-STRESS; STEADY FLOW; BLOOD-FLOW; GUIDE-WIRE; CATHETER; VELOCITY; MODELS AB To evaluate the local hemodynamic implications of coronary artery balloon angioplasty computational fluid dynamics (CFD) was applied in a group Of patients previously reported by [Wilson et al. (1988), 77, pp. 873-885] with representative stenosis geometry post-angioplasty and with measured values of coronary flow reserve returning to a normal range (3.6+/-0.3). During undisturbed flow in the absence of diagnostic catheter sensors within the lesions the computed mean pressure drop Delta(p) over tilde was only about 1 mmHg at basal flow and increased moderately to about 8 mmHg for hyperemic flow. Corresponding elevated levels of mean wall shear stress in the midthroat region of the residual stenoses, which are common after angioplasty procedures, increased from about 60 to 290 dynes/cm(2) during hyperemia. The computations ((R) over bare(e) similar or equal to 100-400; alpha (e) = 2.25) indicated that the pulsatile flow field was principally quasi-steady during the cardiac cycle but there was phase lag in the pressure drop-mean velocity (Deltap - (u) over bar) relation. Time-averaged pressure drop values, Delta(p) over tilde, were about 20 percent higher than calculated pressure drop values, Deltap(s), for steady flow, similar to previous in vitro measurements by Cho et al. (1983). In the throat region, viscous effects were confined to the near-wall region and entrance effects were evident during the cardiac cycle. Proximal to the lesion, velocity profiles deviated from parabolic shape at lower velocities during the cardiac cycle. The flow field was very complex in the oscillatory separated flow reattachment region in the distal vessel where pressure recovery occurred These results may also serve as a useful reference against catheter-measured pressure drops and velocity ratios (hemodynamic endpoints) and arteriographic (anatomic) endpoints post-angioplasty. Some comparisons to previous studies of flow through stenoses models are also shown for perspective purposes. [S0148-0731(00)00304-6]. C1 NIH, Bioengn & Phys Sci Program, Bethesda, MD 20892 USA. CALTECH, Jet Prop Lab, Pasadena, CA 91109 USA. Univ S Florida, Coll Med, Div Vasc Surg, Tampa, FL 33606 USA. Drexel Univ, Dept Mech Engn & Mech, Philadelphia, PA 19104 USA. RP Banerjee, RK (reprint author), NIH, Bioengn & Phys Sci Program, Bldg 3,Rm 3N17, Bethesda, MD 20892 USA. NR 35 TC 24 Z9 27 U1 0 U2 3 PU ASME-AMER SOC MECHANICAL ENG PI NEW YORK PA THREE PARK AVE, NEW YORK, NY 10016-5990 USA SN 0148-0731 J9 J BIOMECH ENG-T ASME JI J. Biomech. Eng.-Trans. ASME PD AUG PY 2000 VL 122 IS 4 BP 310 EP 320 DI 10.1115/1.1287157 PG 11 WC Biophysics; Engineering, Biomedical SC Biophysics; Engineering GA 404QC UT WOS:000167111100003 PM 11036553 ER PT J AU Skrtic, D Antonucci, JM Eanes, ED Eichmiller, FC Schumacher, GE AF Skrtic, D Antonucci, JM Eanes, ED Eichmiller, FC Schumacher, GE TI Physicochemical evaluation of bioactive polymeric composites based on hybrid amorphous calcium phosphates SO JOURNAL OF BIOMEDICAL MATERIALS RESEARCH LA English DT Article DE composites; amorphous calcium phosphate; remineralization; mechanical strength ID PHOSPHATE/METHACRYLATE COMPOSITES AB Amorphous calcium phosphate (ACP)-filled methacrylate composites were recently found to effectively remineralize in vitro caries-like enamel lesions. Their inferior mechanical properties compared to glass-filled composites, however, limit their use as a dental restorative material. In this study, the feasibility of introducing glass-forming elements (tetraethoxysilane or zirconyl chloride) during the low-temperature synthesis of ACP was investigated. Composites based on such hybrid fillers (mass fraction, 40%) were evaluated to establish whether hybridization strengthened the composites, ia improved interfacial interactions with the polymer phase without compromising the release of the mineral ions. Two types of visible-light cured resins were prepared: BTHZ resin from 2,2-bis[p-(2'-hydroxy-3'-methacryloxypropoxy)phenyl] propane (BisGMA), triethylene glycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA) and zirconyl methacrylate (ZrM), and TIP resin front TEGDMA and pyromellitic glycerol dimethacrylate (PMGDM). Hybridized fillers and BTHZ- and TP-based composites were characterized by the IR spectroscopy, X-ray diffraction, dissolution/transformation kinetic studies, and biaxial flexure strength (BFS) testing before and after immersion in buffered saline solutions. The feasibility of improving the BFS via hybridization, while retaining, if not enhancing the remineralizing potential was demonstrated for BTHZ-based composites. Both BFS and remineralizing ability of the TP-composites, however, deteriorated upon their exposure to an aqueous environment. Therefore, hybridized ACP-filled BTHZ composites have a potential for utilization in more demanding restorative, sealant, and adhesive applications. (C) 2000 John Wiley & Sons. Inc.*. C1 Natl Inst Stand & Technol, Amer Dent Assoc Hlth Fdn, Paffenbarger Res Ctr, Gaithersburg, MD 20899 USA. Natl Inst Stand & Technol, Dent & Med Mat Grp, Div Polymer, Gaithersburg, MD 20899 USA. Natl Inst Dent & Craniofacial Res, Mineral Chem & Struct Sect, NIH, Bethesda, MD USA. RP Skrtic, D (reprint author), Natl Inst Stand & Technol, Amer Dent Assoc Hlth Fdn, Paffenbarger Res Ctr, 100 Bur Dr,Stop 8546, Gaithersburg, MD 20899 USA. NR 12 TC 77 Z9 78 U1 2 U2 15 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0021-9304 J9 J BIOMED MATER RES JI J. Biomed. Mater. Res. PD AUG PY 2000 VL 53 IS 4 BP 381 EP 391 DI 10.1002/1097-4636(2000)53:4<381::AID-JBM12>3.0.CO;2-H PG 11 WC Engineering, Biomedical; Materials Science, Biomaterials SC Engineering; Materials Science GA 336MU UT WOS:000088307500012 PM 10898879 ER PT J AU Park, JI Jeong, JS Han, JY Kim, DI Gao, YH Park, SC Rodgers, GP Kim, IH AF Park, JI Jeong, JS Han, JY Kim, DI Gao, YH Park, SC Rodgers, GP Kim, IH TI Hydroxyurea induces a senescence-like change of K562 human erythroleukemia cell SO JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY LA English DT Article DE hydroxyurea; senescence; erythroleukemia K562 cells ID HUMAN-DIPLOID FIBROBLASTS; EXPRESSION; DNA; DIFFERENTIATION; PROTEIN; MARKER; GENES AB Hydroxyurea, a differentiation-inducing agent of human erythroleukemia K562 cells, is commonly used to treat some types of leukemia. However, the mechanism for its therapeutic effect is not clearly understood yet. In this study, we have observed an interesting effect of hydroxyurea on tumor cells: an induction of senescence-like changes. Human erythroleukemia K562 cells, when treated with hydroxyurea for 7 days or more. underwent a change into phenotypically senescent cells together with a reduction of hemoglobin generation, a differentiation marker. The hydroxyurea-treated cells showed positive senescence associated-beta-galactosidase staining, a senescence index, and the accumulation of cdk (cyclin dependent kinase) inhibitors, such as p16(INK4a), p21(Waf1), and p27(Kip1), implicated in cellular senescence. Nonetheless, these changes were not accompanied by DNA fragmentation. Taken together, we summarize that the long-term treatment of cancer cells with hydroxyurea can induce cellular senescence different from differentiation or programmed cell death. C1 Dong A Univ, Coll Med, Dept Biochem, Pusan, South Korea. Dong A Univ, Coll Med, Dept Pathol, Pusan, South Korea. Dong A Univ, Coll Med, Dept Clin Pathol, Pusan, South Korea. Seoul Natl Univ, Coll Med, WHO, Collaborating Ctr Phys Culture & Aging Res Hlth P, Seoul, South Korea. NIDDK, Mol & Clin Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Park, JI (reprint author), Dong A Univ, Coll Med, Dept Biochem, Pusan, South Korea. NR 29 TC 25 Z9 26 U1 1 U2 6 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0171-5216 J9 J CANCER RES CLIN JI J. Cancer Res. Clin. Oncol. PD AUG PY 2000 VL 126 IS 8 BP 455 EP 460 DI 10.1007/s004320050013 PG 6 WC Oncology SC Oncology GA 339DD UT WOS:000088461600004 PM 10961388 ER PT J AU Li, ZH Nater, C Kinsella, J Chrest, F Lakatta, EG AF Li, ZH Nater, C Kinsella, J Chrest, F Lakatta, EG TI Minoxidil inhibits proliferation and migration of cultured vascular smooth muscle cells and neointimal formation after balloon catheter injury SO JOURNAL OF CARDIOVASCULAR PHARMACOLOGY LA English DT Article DE Smooth muscle cell; minoxidil; neointima; migration ID DIRECTIONAL CORONARY ATHERECTOMY; SYMPATHETIC ACTIVITY; AORTIC EXPLANTS; RESTENOSIS; INVITRO; RATS; VASODILATION; ANGIOPLASTY; FATE AB The goal of the study was to investigate the in vitro and in vivo inhibition of minoxidil on smooth muscle cell (SMC) proliferation and migration as well as neointimal formation. The in vitro effect of minoxidil was investigated by Boyden chamber assay and cell-cycle analysis. To evaluate the in vivo effect, we treated the animals with minoxidil in their drinking water before and after balloon catheter injury to carotid artery. Results showed that minoxidil inhibited SMC migration across type I collagen membrane in a dose-related manner (13.5% by 0.01 mg/ml; p < 0.05; 16.8% by 0.05 mg/ml; p < 0.01; 40.4% by 0.25 mg/ml: p < 0.001; and 65.8% by 1.25 mg/ml; p < 0.001). Minoxidil (0.8 mg/ml) increased the number of SMCs in G(1) phase (p < 0.05) and decreased the number of SMCs in S phase (p < 0.001). In vivo minoxidil treatment reduced neointimal mass by 31.7% (120 mg/L) and 42.3% (200 mg/L), respectively. Data demonstrate that minoxidil inhibits vascular SMC proliferation and migration both in vitro and in vivo, and therefore may be useful to inhibit SMC hyperplasia that occurs in restenosis and other vascular diseases. C1 NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Li, ZH (reprint author), CV Therapeut, Dept Pharmacol Sci, 3172 Porter Dr, Palo Alto, CA 94304 USA. NR 32 TC 3 Z9 3 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0160-2446 J9 J CARDIOVASC PHARM JI J. Cardiovasc. Pharmacol. PD AUG PY 2000 VL 36 IS 2 BP 270 EP 276 DI 10.1097/00005344-200008000-00019 PG 7 WC Cardiac & Cardiovascular Systems; Pharmacology & Pharmacy SC Cardiovascular System & Cardiology; Pharmacology & Pharmacy GA 339CK UT WOS:000088459900019 PM 10942171 ER PT J AU Bogan, JA Natale, DA Depamphilis, ML AF Bogan, JA Natale, DA Depamphilis, ML TI Initiation of eukaryotic DNA replication: Conservative or liberal? SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Review ID ORIGIN RECOGNITION COMPLEX; CHORION GENE AMPLIFICATION; XENOPUS EGG EXTRACTS; BETA-GLOBIN LOCUS; CELL-FREE SYSTEM; S-PHASE; FISSION YEAST; DROSOPHILA-MELANOGASTER; AUTONOMOUS REPLICATION; CDK PHOSPHORYLATION AB The mechanism for initiation of eukaryotic DNA replication is highly conserved: the proteins required to initiate replication, the sequence of events leading to initiation, and the regulation of initiation are remarkably similar throughput the eukaryotic kingdom. Nevertheless, there is a liberal attitude when it comes to selecting initiation sites. Differences appear to exist in the composition of replication origins and in the way proteins recognize these origins. In fact, some multicellular eukaryotes (the metazoans) can change the number and locations of initiation sites during animal development, revealing that selection of initiation sites depends on epigenetic as well as genetic parameters. Here we have attempted to summarize our understanding of this process, to identify the similarities and differences between single cell and multicellular eukaryotes, and to examine the extent to which origin recognition proteins and replication origins have been conserved among eukaryotes. J. Cell. Physiol. 184:139-150, 2000. Published 2000 Wiley-Liss, Inc.dagger C1 NICHHD, NIH, Bethesda, MD 20892 USA. RP Bogan, JA (reprint author), NICHHD, NIH, Bldg 6,Room 416, Bethesda, MD 20892 USA. NR 83 TC 69 Z9 71 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 0021-9541 J9 J CELL PHYSIOL JI J. Cell. Physiol. PD AUG PY 2000 VL 184 IS 2 BP 139 EP 150 DI 10.1002/1097-4652(200008)184:2<139::AID-JCP1>3.0.CO;2-8 PG 12 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 330AC UT WOS:000087939800001 PM 10867638 ER PT J AU Yanovski, JA Yanovski, SZ Boyle, AJ Gold, PW Sovik, KN Sebring, NG Drinkard, B AF Yanovski, JA Yanovski, SZ Boyle, AJ Gold, PW Sovik, KN Sebring, NG Drinkard, B TI Hypothalamic-pituitary-adrenal axis activity during exercise in African American and Caucasian women SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID CORTICOTROPIN-RELEASING-FACTOR; WHITE WOMEN; WEIGHT-LOSS; ARGININE-VASOPRESSIN; FRAMESHIFT MUTATION; DOSE-RESPONSE; UNITED-STATES; OBESITY; ADRENOCORTICOTROPIN; SECRETION AB African American women have a greater prevalence of obesity than Caucasian women, but the reasons for this difference are not known. We have investigated whether activity of the hypothalamic-pituitary adrenal axis plays a role in this phenomenon. Previous studies have shown that plasma ACTH immunoreactivity (ACTH-IR) of African American women, measured after ovine CRH (oCRH) stimulation, is significantly greater than ACTH-IR of Caucasian women, but is not accompanied by greater plasma cortisol concentrations. Analysis by high pressure liquid chromatography has demonstrated that after oCRH stimulation, the plasma ACTH-IR of African American women contains many nonintact ACTH fragments not found in Caucasians. To determine whether these racial differences in ACTH-IR secretion are an artifact of exogenous oCRH administration or are also found after a physiological stimulus for ACTH secretion, we measured hormones of the hypothalamic-pituitary adrenal axis before and after a standardized, maximal exercise treadmill test in 16 African American and 19 Caucasian healthy women matched for age, socioeconomic status, and body mass index. The intensity of exercise performed was similar in the two groups, as determined by duration of exercise, perceived intensity of exertion, plasma lactate, maximal heart rate, and maximum oxygen uptake. Basal ACTH-IR measured by RIA or immunoradiometric assay and cortisol were similar in African Americans and Caucasians. Plasma ACTH-IR, measured 10 min after completion of exercise, was significantly greater in African Americans than in Caucasians [by RIA: mean +/- so ACTH-IR, 47.1 +/- 30.9 vs. 25.4 +/- 16.7 pmol/ (P < 0.01); by immunoradiometric assay: ACTH-IR, 45.9 +/- 43.2 vs. 21.1 +/- 14.6 pmol/L (P < 0.05)]. However, plasma cortisol after exercise was not different (450.2 +/- 157.7 vs. 483.6 +/- 180.4 nmol/L; P = 0.57). We conclude that ACTH-IR is significantly greater in African American than in Caucasian women after intense exercise. The ACTH-IR of African Americans and Caucasians does not appear to be equipotent at adrenal melanocortin-2 receptors, because the greater ACTH-IR of African Americans does not lead to greater cortisol secretion. Whether some components of the ACTH-IR detected in African Americans affect signal transduction of the hypothalamic melanocortin-4 receptors implicated in body weight regulation and thus predispose African American women to weight gain without altering plasma cortisol remains to be determined. C1 NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NIMH, Clin Neuroendocrinol Branch, NIH, Bethesda, MD 20892 USA. NIDDK, Div Digest Dis & Nutr, NIH, Bethesda, MD 20892 USA. NIDDK, Div Nutr Res Coordinat, NIH, Bethesda, MD 20892 USA. NIH, Dept Nutr, Bethesda, MD 20892 USA. NIH, Dept Rehabil Med, Bethesda, MD 20892 USA. NIH, Warren Grant Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Yanovski, JA (reprint author), NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, 10 Ctr Dr,MSC 1862,Bldg 10,Room 10N262, Bethesda, MD 20892 USA. EM JYISi@NIH.Gov OI Yanovski, Jack/0000-0001-8542-1637 FU NICHD NIH HHS [Z01-HD-00641] NR 29 TC 17 Z9 17 U1 0 U2 3 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD AUG PY 2000 VL 85 IS 8 BP 2660 EP 2663 DI 10.1210/jc.85.8.2660 PG 4 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 381VQ UT WOS:000165784500005 PM 10946862 ER PT J AU Falkeborn, M Schairer, C Naessen, T Persson, I AF Falkeborn, M Schairer, C Naessen, T Persson, I TI Risk of myocardial infarction after oophorectomy and hysterectomy SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article DE hysterectomy; oophorectomy; acute myocardial infarction; case cohort study; natural menopause; myoma ID CORONARY HEART-DISEASE; PREMENOPAUSAL HYSTERECTOMY; CARDIOVASCULAR MORTALITY; REPRODUCTIVE FACTORS; UTERINE MYOMAS; WOMEN; MENOPAUSE; AGE; PROGESTOGEN; ESTROGEN AB To determine the risk of developing a first myocardial infarction after a hysterectomy and/or oophorectomy. Case-cohort analysis performed among 17,126 women in the Uppsala Health Care Region of Sweden, who had undergone a hysterectomy and/or oophorectomy in 1965 to 1983. Record linkage was used for follow-up and medical records to ascertain the actual history of oophorectomy. Risk estimates were calculated by relating the observed number of cases in the cohort to that expected on the basis of incidence rates in the population. Overall, 214 cases of myocardial infarction were observed. In premenopausal women a bilateral oophorectomy alone tended to increase the relative risk 1.6; 95% CI 0.8-3.1, but this operation combined with hysterectomy increased the risk only among those aged 50 and over at surgery. Hysterectomy at premenopausal age or unilateral oophorectomy did not alter the risk of myocardial infarction. In naturally menopausal women, hysterectomy-mainly for uterine myoma-was associated with a four-fold increase in relative risk (3.8; 95% CI 1.9-7.8). Hysterectomy for treatment of myoma performed after a natural menopause is linked to an excess risk for myocardial infarction. Bilateral oophorectomy before menopause may increase the risk of myocardial infarction. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Uppsala, Dept Publ Hlth & Caring Sci Geriatr, Uppsala, Sweden. NCI, Environm Epidemiol Branch, Rockville, MD USA. Univ Uppsala Hosp, Dept Gynecol & Obstet, Uppsala, Sweden. Karolinska Inst, Dept Med Epidemiol, Stockholm, Sweden. RP Falkeborn, M (reprint author), Dept Geriatr, POB 609, S-75125 Uppsala, Sweden. NR 25 TC 54 Z9 57 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD AUG PY 2000 VL 53 IS 8 BP 832 EP 837 DI 10.1016/S0895-4356(00)00187-6 PG 6 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 347CA UT WOS:000088908400009 PM 10942866 ER PT J AU Engels, EA Terrin, N Barza, M Lau, J AF Engels, EA Terrin, N Barza, M Lau, J TI Meta-analysis of diagnostic tests for acute sinusitis SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article DE sinusitis; diagnostic tests; meta-analysis; receiver operator curve; radiography; clinical examination ID ACUTE MAXILLARY SINUSITIS; PLACEBO-CONTROLLED TRIAL; PHYSICAL-EXAMINATION; ULTRASONOGRAPHY; RADIOGRAPHY; MANAGEMENT; THERAPY; HISTORY AB To facilitate management of acute sinusitis, we conducted a meta-analysis of published studies comparing diagnostic tests for this disorder. Thirteen studies were identified through literature search. Based on sinus puncture/aspiration (considered most accurate), 49-83 % of symptomatic patients had acute sinusitis. Compared with puncture/aspiration, radiography offered moderate ability to diagnose sinusitis (summary receiver operator curve [SROC] area, 0.83). Using sinus opacity or fluid as the criterion for sinusitis, radiography had sensitivity of 0.73 and specificity of 0.80. Studies evaluating ultrasonography revealed substantial variation in test performance. The clinical evaluation, particularly risk scores formally incorporating history and physical examination findings, had moderate ability to identify patients with positive radiographs (SROC area, 0.74). Many studies were of poor quality, with inadequately described test methods and unblinded test interpretation. In conclusion, acute sinusitis is common among symptomatic patients. Radiography and clinical evaluation (especially risk scores) appear to provide useful information for diagnosis of sinusitis. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Tufts Univ, Sch Med, New England Med Ctr, Dept Med,Div Clin Care Res, Boston, MA 02111 USA. Tufts Univ, Sch Med, New England Med Ctr, Dept Med,Div Geog Med & Infect Dis, Boston, MA 02111 USA. Tufts Univ, Sch Med, New England Med Ctr, Tupper Res Inst, Boston, MA 02111 USA. RP Engels, EA (reprint author), NCI, Div Canc Epidemiol & Genet, Viral Epidemiol Branch, 6120 Execut Blvd,MSC 7248, Rockville, MD 20822 USA. FU AHRQ HHS [T32HS00060, R25HS09796] NR 33 TC 60 Z9 62 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD AUG PY 2000 VL 53 IS 8 BP 852 EP 862 DI 10.1016/S0895-4356(00)00209-2 PG 11 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 347CA UT WOS:000088908400012 PM 10942869 ER PT J AU Kulldorff, M McShane, LM Schatzkin, A Freedman, LS Wargovich, MJ Woods, C Purewal, M Burt, RW Lawson, M Mateski, DJ Lanza, E Corle, DK O'Brien, B Moler, J AF Kulldorff, M McShane, LM Schatzkin, A Freedman, LS Wargovich, MJ Woods, C Purewal, M Burt, RW Lawson, M Mateski, DJ Lanza, E Corle, DK O'Brien, B Moler, J TI Measuring cell proliferation in the rectal mucosa: comparing bromodeoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA) assays SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article DE colorectal cancer; colon adenomas; correlation; variance components; biopsies ID POLYP PREVENTION TRIAL; COLON-CANCER RISK; COLORECTAL MUCOSA; CALCIUM SUPPLEMENTATION; 5-BROMO-2-DEOXYURIDINE; ADENOMA; INVITRO; INVIVO AB Cell proliferation in the human colorectum can be measured using bromodeoxyuridine (BrdU) or proliferating cell nuclear antigen (PCNA) assays. Using data from the National Cancer Institute's Polyp Prevention Trial, these two assays are compared using correlation coefficients and variance components analysis. Adjusting for fixed as well as for the random effects of between-biopsy and scoring variation, the estimated correlation is 0.46 for the log labeling index and 0.45 for log proliferative height. This is an estimate of the highest correlation that can be achieved by taking multiple biopsies scored by multiple scorers. For single biopsies, the estimated correlation is 0.16 and 0.10, respectively. There are significant differences between the variance components for the two assays. For example, for log labeling index, PCNA has a lower variation between biopsies than BrdU, but higher variation between scorings, When used in a clinical or epidemiological setting, it is important to take multiple biopsies at multiple time points. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NCI, Bethesda, MD 20892 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Univ Utah, Salt Lake City, UT USA. Kaiser Permanente Med Ctr, Sacramento, CA USA. Walter Reed Army Med Ctr, Washington, DC 20307 USA. Westat Inc, Rockville, MD USA. Informat Management Serv, Rockville, MD USA. RP Kulldorff, M (reprint author), Univ Connecticut, Sch Med, Dept Community Med, Div Biostat, Farmington, CT 06030 USA. RI Kulldorff, Martin/H-4282-2011; OI Kulldorff, Martin/0000-0002-5284-2993; Moler, James/0000-0001-8738-6898 NR 37 TC 8 Z9 13 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD AUG PY 2000 VL 53 IS 8 BP 875 EP 883 DI 10.1016/S0895-4356(99)00180-8 PG 9 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 347CA UT WOS:000088908400015 PM 10942872 ER PT J AU Erice, A Brambilla, D Bremer, J Jackson, JB Kokka, R Yen-Lieberman, B Coombs, RW AF Erice, A Brambilla, D Bremer, J Jackson, JB Kokka, R Yen-Lieberman, B Coombs, RW TI Performance characteristics of the QUANTIPLEX HIV-1 RNA 3.0 assay for detection and quantitation of human immunodeficiency virus type 1 RNA in plasma SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID ACTIVE ANTIRETROVIRAL THERAPY; VIRAL LOAD; MONITOR ASSAY; INFECTION; QUANTIFICATION; TRANSCRIPTION; PROGRESSION AB The QUANTIPLEX HIV-1 RNA assay, version 3.0 (a branched DNA, version 3.0, assay [bDNA 3.0 assay]), was evaluated by analyzing spiked and clinical plasma samples and was compared with the AMPLICOR HIV-1 MONITOR Ultrasensitive (ultrasensitive reverse transcription-PCR [US-RT-PCR]) method. A panel of spiked plasma samples that contained 0 to 750,000 copies of human immunodeficiency virus type 1 (HIV-1) RNA per ml was tested four times in each of four laboratories (1,344 assays). Negative results (<50 copies/ml) were obtained in 30 of 32 (94%) assays with seronegative samples, 66 of 128 (52%) assays with HIV-1 RNA at 50 copies/ml, and 5 of 128 (4%) assays with HIV-1 RNA at 100 copies/ml. The assay was linear from 100 to 500,000 copies/ml. The within-run standard deviation (SD) of the log(10) estimated HIV-1 RNA concentration was 0.08 at 1,000 to 500,000 copies/ml, increased below 1,000 copies/ml, and was 0.17 at 100 copies/ml. Between-run reproducibility at 100 to 500 copies/ml was <0.10 log(10) in most comparisons. Interlaboratory differences across runs were less than or equal to 0.10 log(10) at all concentrations examined. A subset of the panel (25 to 500 copies/ml) was also analyzed by the US-RT-PCR assay. The within-run SD varied inversely with the log(10) HIV-1 RNA concentration but was higher than the SD for the bDNA 3.0 assay at all concentrations. Log-log regression analysis indicated that the two methods produced very similar estimates at 100 to 500 copies/ml. In parallel testing of clinical specimens with low HIV-1 RNA levels, 80 plasma samples with <50 copies/ml by the US-RT-PCR assay had <50 copies/ml when they were retested by the bDNA 3.0 assay. In contrast, 11 of 78 (14%) plasma samples with <50 copies/ml by the bDNA 3.0 assay had greater than or equal to 50 copies/ml when they were retested by the US-RT-PCR assay (median, 86 copies/ml; range, 50 to 217 copies/ml). Estimation of bDNA 3.0 values of <50 copies/ml by extending the standard curve of the assay showed that these samples with discrepant results had higher HIV-1 RNA levels than the samples with concordant results (median, 34 versus 17 copies/ml; P = 0.0051 by the Wilcoxon two-sample test). The excellent reproducibility, broad linear range, and good sensitivity of the bDNA 3.0 assay make it a very attractive method for quantitation of HIV-1 RNA levels in plasma. C1 Univ Minnesota, Dept Lab Med & Pathol, Minneapolis, MN 55455 USA. Univ Minnesota, Dept Med, Div Infect Dis, Minneapolis, MN 55455 USA. New England Res Inst, Watertown, MA 02172 USA. Rush Presbyterian St Lukes Med Ctr, Chicago, IL 60612 USA. Johns Hopkins Univ, Dept Pathol, Baltimore, MD USA. Bayer Nucle Acid Diagnost, Emeryville, CA USA. Cleveland Clin, Dept Clin Pathol, Cleveland, OH USA. Univ Washington, Dept Lab Med, Seattle, WA 98195 USA. NIAID, Div Aids, Virol Qual Assurance Lab Program, Bethesda, MD 20892 USA. RP Erice, A (reprint author), Univ Minnesota, Dept Lab Med & Pathol, Box 437 Mayo,420 Delaware St SE, Minneapolis, MN 55455 USA. FU NIAID NIH HHS [P01 AI030731, AI2776, AI27664, U01 AI027664] NR 23 TC 33 Z9 34 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 2000 VL 38 IS 8 BP 2837 EP 2845 PG 9 WC Microbiology SC Microbiology GA 340XL UT WOS:000088561700006 PM 10921936 ER PT J AU Rugge, M Shiao, YH Busatto, G Cassaro, M Strobbe, C Russo, VM Leo, G Parenti, AR Scapinello, A Arslan, P Egarter-Vigl, E AF Rugge, M Shiao, YH Busatto, G Cassaro, M Strobbe, C Russo, VM Leo, G Parenti, AR Scapinello, A Arslan, P Egarter-Vigl, E TI The p53 gene in patients under the age of 40 with gastric cancer: mutation rates are low but are associated with a cardiac location SO JOURNAL OF CLINICAL PATHOLOGY-MOLECULAR PATHOLOGY LA English DT Article DE stomach neoplasm; genes; p53 ID ESOPHAGEAL CANCER; CARCINOMA; PHENOTYPE; 5-METHYLCYTOSINE; ADENOCARCINOMAS; ETIOLOGY; PROTEIN; TUMORS; REGION AB Background-Determining both the frequency and the spectrum of p53 gene mutation in young patients with gastric cancer might provide clues to the host related genetic mechanism(s) in gastric carcinogenesis. Patients and methods-p53 mutations were assessed (by means of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), followed by DNA sequencing) in a cohort of 105 consecutive Italian patients in whom gastric cancer was ascertained before the age of 41. Results-A low prevalence of p53 mutations (eight of 105) was observed, with no significant difference between intestinal (three of 31; 10%) and diffuse (five of 74; 7%) phenotypes. A significantly higher prevalence of p53 mutations was associated with the cardiac location (odds ratio, 7.09; confidence interval, 1.56 to 32.11). In all but one case, p53 mutations were associated with a stage higher than I. All eight mutations were located at CpG sites, where G : C to A: T transitions have been associated with frequent methylation at the C5 position of cytosine. Conclusions-These findings show that, unlike what has been consistently demonstrated in the general population, p53 mutations are uncommon in gastric cancer occurring in young patients, and in such patients, p53 alterations are significantly associated with the cardiac location. C1 Univ Padua, Dept Oncol & Surg Sci, ULSS15 Reg Veneto, I-35121 Padua, Italy. NCI, Comparat Carcinogenesis Lab, FCRDC, NIH, Frederick, MD 21702 USA. Castelfranco Veneto Hosp, Dept Pathol, I-31033 Bassano, Italy. Univ Padua, Dept Med & Diagnost Sci, I-35121 Padua, Italy. Bolzano Gen Hosp, Dept Pathol, I-39100 Bolzano, Italy. RP Rugge, M (reprint author), Univ Padua, Dept Oncol & Surg Sci, ULSS15 Reg Veneto, Via A Gabelli 61, I-35121 Padua, Italy. RI Rugge, Massimo/K-7525-2016 NR 30 TC 20 Z9 20 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 1366-8714 J9 J CLIN PATHOL-MOL PA JI J. Clin. Pathol.-Mol. Pathol. PD AUG PY 2000 VL 53 IS 4 BP 207 EP 210 PG 4 WC Pathology SC Pathology GA 341XP UT WOS:000088616500008 ER PT J AU Field, MS Pinsky, PF AF Field, MS Pinsky, PF TI A two-region nonequilibrium model for solute transport in solution conduits in karstic aquifers SO JOURNAL OF CONTAMINANT HYDROLOGY LA English DT Article DE karstic aquifers; solution conduits; nonequilibrium modeling; velocity; longitudinal dispersion ID SORBING POROUS-MEDIA; LONGITUDINAL DISPERSION; STORAGE MODEL; MASS-TRANSFER; FLOW; FRACTURE; EXCHANGE AB A two-region nonequilibrium model was used to calibrate initial solute-transport parameter estimates generated from tracer-breakthrough curves (TBCs) developed from tracer tests conducted in uni-axial solution conduits in karstic aquifers. Two-region nonequilibrium models account for partitioning of solute into mobile- and immobile-fluid regions to produce a more representative model fit to the strong tails associated with TBCs than do equilibrium models. The nonequilibrium model resulted in an increase in average flow velocities and a decrease in longitudinal dispersion coefficients over comparable estimates using an equilibrium model. Increases in velocity and decreases in dispersion were obtained at the expense of including parameters that describe solute partitioning and mass transfer rate for the mobile- and immobile-fluid regions. In addition, nonidentifiable sorption and mass transfer parameters for the immobile-fluid regions could only be described in terms of upper and lower bounds using readily determined identifiable ratios representing solute partitioning and system constraints based on known physical properties. The identifiable ratios and system constraints serve to minimize model nonuniqueness and renders the nonidentification problem trivial. Published by Elsevier Science B.V. C1 US EPA, Natl Ctr Environm Assessment, Washington, DC 20460 USA. NCI, NIH, Rockville, MD USA. RP Field, MS (reprint author), US EPA, Natl Ctr Environm Assessment, 401 M St SW, Washington, DC 20460 USA. OI Field, Malcolm/0000-0002-8350-417X NR 55 TC 67 Z9 70 U1 1 U2 12 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-7722 J9 J CONTAM HYDROL JI J. Contam. Hydrol. PD AUG PY 2000 VL 44 IS 3-4 BP 329 EP 351 DI 10.1016/S0169-7722(00)00099-1 PG 23 WC Environmental Sciences; Geosciences, Multidisciplinary; Water Resources SC Environmental Sciences & Ecology; Geology; Water Resources GA 350YD UT WOS:000089128500006 ER PT J AU Wisbeck, JM Huffman, LC Freund, L Gunnar, MR Davis, EP Reiss, AL AF Wisbeck, JM Huffman, LC Freund, L Gunnar, MR Davis, EP Reiss, AL TI Cortisol and social stressors in children with fragile X: A pilot study SO JOURNAL OF DEVELOPMENTAL AND BEHAVIORAL PEDIATRICS LA English DT Article DE fragile X syndrome; salivary cortisol; social stressors; neuroendocrine; hippocampus ID COGNITIVE DEFICITS; SALIVARY CORTISOL; BEHAVIOR PROBLEMS; FRA(X) SYNDROME; MALES; RESPONSES; GLUCOCORTICOIDS; HIPPOCAMPUS; REACTIVITY; PHENOTYPE AB Evidence of neuroendocrine dysfunction, behavioral features of social anxiety and avoidance, and neuroanatomical abnormalities suggest that abnormal hypothalamic-pituitary-adrenal (HPA) function may be a component of the fragile X (fra X) syndrome. In this preliminary study, salivary cortisol levels of males (n = 8, mean age = 13.5 yr) and females (n = 7, mean age = 13.9 yr) with the fra X full mutation were studied for 3 days. Day 1 was an experimental day, during which subjects experienced a Social Stressor task midmorning. Days 2 and 3 were routine days, during which the subjects were engaged in their typical activities. Saliva samples were collected before breakfast, lunch, dinner, and bedtime. On the experimental day, the prelunch sample collection occurred 30 and 90 minutes after the Social Stressor task. Compared with children's norms, the combined group of mates and females with fra X had significantly higher cortisol levels in the prelunch and the prebedtime samples for the routine days. Comparisons between the two fra X groups for the experimental day revealed similar diurnal patterns for cortisol level. However, compared with females with fra X, males with fra X had significantly higher cortisol levels at two points during the day: 30 minutes after the social stressor and at bedtime. These preliminary data suggest that individuals with fra X have abnormal HPA function. Understanding the relations among HPA dysfunction, abnormalities in brain structure and/or function, and maladaptive behavior and cognition in fra X could inform the design of early interventions using pharmacological or environmental measures designed to normalize neuroendocrine function. J Dev Behav Pediatr 21:278-282 2000. Index terms: fragile X syndrome, salivary cortisol, social stressors, neuroendocrine, hippocampus. C1 Stanford Univ, Sch Med, Dept Pediat, Stanford, CA 94305 USA. Childrens Hlt Council, Palo Alto, CA USA. Stanford Univ, Sch Med, Behav Neurogenet Res Ctr, Stanford, CA 94305 USA. NICHHD, Bethesda, MD 20892 USA. Univ Minnesota, Inst Child Dev, Minneapolis, MN 55455 USA. Stanford Univ, Sch Med, Dept Psychiat & Behav Sci, Stanford, CA 94305 USA. RP Huffman, LC (reprint author), Stanford Univ, Sch Med, 700 Sand Hill Rd, Palo Alto, CA 94304 USA. RI Davis, Elysia/B-7621-2013 NR 39 TC 40 Z9 41 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0196-206X J9 J DEV BEHAV PEDIATR JI J. Dev. Behav. Pediatr. PD AUG PY 2000 VL 21 IS 4 BP 278 EP 282 DI 10.1097/00004703-200008000-00004 PG 5 WC Behavioral Sciences; Psychology, Developmental; Pediatrics SC Behavioral Sciences; Psychology; Pediatrics GA 347ED UT WOS:000088913700004 PM 10972251 ER PT J AU Walsh, PJ Heitz, MJ Campbell, CE Cooper, GJ Medina, M Wang, YXS Goss, GG Vincek, V Wood, CM Smith, CP AF Walsh, PJ Heitz, MJ Campbell, CE Cooper, GJ Medina, M Wang, YXS Goss, GG Vincek, V Wood, CM Smith, CP TI Molecular characterization of a urea transporter in the gill of the gulf toadfish (Opsanus beta) SO JOURNAL OF EXPERIMENTAL BIOLOGY LA English DT Article DE gulf toadfish; Opsanus beta; urea transporter gene; UT-A2; gill; nitrogen excretion ID EARLY-LIFE STAGES; NITROGEN-EXCRETION; CARDIORESPIRATORY PHYSIOLOGY; FUNCTIONAL-CHARACTERIZATION; CYCLE ENZYMES; CLONING; EXPRESSION; FISHES; METABOLISM; ELASMOBRANCH AB Urea excretion by the gulf toadfish (Opsanus beta) has been shown in previous studies to be a highly pulsatile facilitated transport, with excretion probably occurring at the gill, The present study reports the isolation of an 1800 base pair (kb) cDNA from toadfish gill with one open reading frame putatively encoding a 475-residue protein, the toadfish urea transporter (tUT). tUT, the first teleostean urea transporter cloned, has high homology with UTs (facilitated urea transporters) cloned from mammals, an amphibian and a shark, and most closely resembles the UT-A subfamily. When expressed in Xenopus laevis oocytes, tUT increased urea permeability (as measured by [C-14]urea uptake) five- to sevenfold, and this permeability increase was abolished by phloretin, a common inhibitor of other UTs, Northern analysis using the 1.8 kb clone was performed to determine the tissue distribution and dynamics of tUT mRNA expression, Of six tissues examined (gill, liver, red blood cells, kidney, skin and intestine), only gill showed expression of tUT mRNA, with a predominant band at 1.8 kb and a minor band at 3.5 kb, During several points in the urea pulse cycle of toadfish (0, 4, 6, 12 and 18 h post-pulse), measured by excretion of [C-14]urea into the water, gill mRNA samples were obtained. Expression of tUT mRNA was found to be largely invariant relative to expression of p-actin mRNA over the pulse cycle. These results further confirm the gill localization of urea transport in the toadfish and suggest that tUT regulation (and the regulation of pulsatile urea excretion) is probably not at the level of mRNA control, The results are discussed in the context of the mechanisms of vasopressin-regulated UT-A in mammalian kidney and morphological data for the toadfish gill. C1 Univ Miami, Rosenstiel Sch Marine & Atmospher Sci, Div Marine Biol & Fisheries, NIEHS,Marine & Freshwater Biomed Sci Ctr, Miami, FL 33149 USA. Univ Manchester, Sch Biol Sci, Manchester M13 9PT, Lancs, England. Univ Alberta, Dept Biol Sci, Edmonton, AB T6G 2E9, Canada. Univ Miami, Sch Med 2E9, Dept Microbiol & Immunol Sci, Miami, FL 33136 USA. Univ Miami, Sch Med 2E9, Dept Pathol, Miami, FL 33136 USA. McMaster Univ, Dept Biol, Hamilton, ON L8S 4K1, Canada. RP Walsh, PJ (reprint author), Univ Miami, Rosenstiel Sch Marine & Atmospher Sci, Div Marine Biol & Fisheries, NIEHS,Marine & Freshwater Biomed Sci Ctr, 4600 Rickenbacker Causeway, Miami, FL 33149 USA. RI Goss, Greg/A-2646-2014; Goss, Greg/D-2723-2015 OI Goss, Greg/0000-0003-0786-8868 NR 34 TC 83 Z9 85 U1 1 U2 3 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 2000 VL 203 IS 15 BP 2357 EP 2364 PG 8 WC Biology SC Life Sciences & Biomedicine - Other Topics GA 345RW UT WOS:000088829800011 PM 10887074 ER PT J AU Zhang, YJ Deng, JH Rabkin, C Gao, SJ AF Zhang, YJ Deng, JH Rabkin, C Gao, SJ TI Hot-spot variations of Kaposi's sarcoma-associated herpesvirus latent nuclear antigen and application in genotyping by PCR-RFLP SO JOURNAL OF GENERAL VIROLOGY LA English DT Article ID MULTICENTRIC CASTLEMANS-DISEASE; EPSTEIN-BARR-VIRUS; PRIMARY EFFUSION LYMPHOMA; CAVITY-BASED LYMPHOMAS; DNA-SEQUENCES; MULTIPLE-MYELOMA; BONE-MARROW; HUMAN-HERPESVIRUS-8 INFECTION; SEXUAL TRANSMISSION; STRAIN VARIABILITY AB Kaposi's sarcoma-associated herpesvirus (KSHV, human herpesvirus-8) is aetiologically associated with Kaposi's sarcoma and several other lymphoproliferative disorders. The latent nuclear antigen (LNA) encoded by KSHV ORF73 has important functions in virus latent infection and shows molecular polymorphism. Sequence variations were identified in the internal repeat domain (IRD) of ORF73. DNA sequencing of ORF73 from one KSHV-infected cell line, PK-1, revealed that there were 558 bp (30.2%) deletions and 66 (3.6%) point mutations located mainly in repeat region 2, the glutamine-rich region of ORF73 IRD, compared with ORF73 of BC-1 KSHV. Similar sequence variations of ORF73 were also identified in two other isolates. None of the sequence variations caused any translational frame-shift in these four KSHV isolates examined, suggesting that LNA has a conservative function in virus latent infection. The frequent sequence variations in repeat region 2 of ORF73 IRD were also identified by PCR-RFLP genotyping in 26 KSHV isolates, suggesting that this region is a 'hot-spot' for genetic variations. Each Kaposi's sarcoma lesion sample contained one virus genotype with a unique RFLP pattern, indicating that in vivo KSHV infection was established with single predominate genotypes, which was further supported by the presence of invariable genotypes in multifocal lesions from individual KS patients. Four KSHV subtypes were classified based on the RFLP patterns that represent the patterns of DNA sequence variations in the ORF73 IRD. PCR-RFLP genotyping is capable of identifying LNA genetic variations and differentiating individual KSHV isolates, and thus may be useful for KSHV molecular epidemiology studies. C1 Univ Texas, Hlth Sci Ctr, Dept Pediat, San Antonio, TX 78229 USA. Univ Texas, Hlth Sci Ctr, Dept Microbiol, San Antonio, TX 78229 USA. NCI, Bethesda, MD 20892 USA. RP Gao, SJ (reprint author), Univ Texas, Hlth Sci Ctr, Dept Pediat, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA. RI Gao, Shou-Jiang/B-8641-2012 FU NHLBI NIH HHS [HL60604] NR 63 TC 17 Z9 20 U1 0 U2 0 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AE, BERKS, ENGLAND SN 0022-1317 J9 J GEN VIROL JI J. Gen. Virol. PD AUG PY 2000 VL 81 BP 2049 EP 2058 PN 8 PG 10 WC Biotechnology & Applied Microbiology; Virology SC Biotechnology & Applied Microbiology; Virology GA 340AF UT WOS:000088509900020 PM 10900044 ER PT J AU Handa, A Muramatsu, S Qiu, JM Mizukami, H Brown, KE AF Handa, A Muramatsu, S Qiu, JM Mizukami, H Brown, KE TI Adeno-associated virus (AAV)-3-based vectors transduce haematopoietic cells not susceptible to transduction with AAV-2-based vectors SO JOURNAL OF GENERAL VIROLOGY LA English DT Article ID HEMATOPOIETIC PROGENITOR CELLS; FIBROBLAST-GROWTH-FACTOR; MEDIATED GENE-TRANSFER; HUMAN PARVOVIRUS B19; CORD BLOOD; EXPRESSION; AAV; INTEGRATION; INFECTION; TYPE-2 AB Although adeno-associated virus (AAV)-2 has a broad tissue-host range and can transduce a wide variety of tissue types, some cells, such as erythro-megakaryoblastoid cells, are non-permissive and appear to lack the AAV-2 receptor. However, limited studies have been reported with the related dependovirus AAV-3, We have previously cloned this virus, characterized its genome and produced an infectious clone. In this study, the gene for green fluorescent protein (GFP) was inserted into AAV-2- and AAV-3-based plasmids and recombinant viruses were produced. These viruses were then used to transduce haematopoietic cells and the transduction efficiencies were compared. In contrast to recombinant (r) AAV-2, rAAV-3 successfully transduced erythroid and megakaryoblastoid cells, although rAAV-2 was superior in transduction of lymphocyte-derived cell lines. Recently, it was reported that heparan sulphate can act as a receptor of AAV-2, The infectivity of rAAV-2 and rAAV-3 was tested with mutant cell lines of Chinese hamster ovary cells that were defective for heparin or heparan sulphate expression on the cell surface. There was no correlation between the ability of rAAV-2 or rAAV-3 to infect cells and the cell surface expression of heparan sulphate and, although heparin blocked both rAAV-2 and rAAV-3 transduction, the ID50 of rAAV-3 was higher than that of rAAV-2. In addition, virus-binding overlay assays indicated that AAV-2 and AAV-3 bound different membrane proteins. These results suggest not only that there are different cellular receptors for AAV-2 and AAV-3, but that rAAV-3 vectors may be preferred for transduction of some haematopoietic cell types. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Handa, A (reprint author), NHLBI, Hematol Branch, NIH, Bldg 10,Rm 7C218,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Mizukami, Hiroaki/D-7674-2013; OI Mizukami, Hiroaki/0000-0001-8954-874X; Muramatsu, Shinichi/0000-0002-3185-7790 NR 30 TC 73 Z9 77 U1 0 U2 2 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AE, BERKS, ENGLAND SN 0022-1317 J9 J GEN VIROL JI J. Gen. Virol. PD AUG PY 2000 VL 81 BP 2077 EP 2084 PN 8 PG 8 WC Biotechnology & Applied Microbiology; Virology SC Biotechnology & Applied Microbiology; Virology GA 340AF UT WOS:000088509900023 PM 10900047 ER PT J AU Rywik, SL Williams, OD Pajak, A Broda, G Davis, CE Kawalec, E Manolio, TA Piotrowski, W Hutchinson, R AF Rywik, SL Williams, OD Pajak, A Broda, G Davis, CE Kawalec, E Manolio, TA Piotrowski, W Hutchinson, R TI Incidence and correlates of hypertension in the Atherosclerosis Risk in Communities (ARIC) study and the Monitoring Trends and Determinants of Cardiovascular Disease (POL-MONICA) project SO JOURNAL OF HYPERTENSION LA English DT Article DE Pol-MONICA; ARIC; hypertension incidence; risk factors ID BLOOD-PRESSURE; ALCOHOL-CONSUMPTION; UNITED-STATES; YOUNG-ADULTS; EPIDEMIOLOGY; POPULATION; PREVALENCE; MANAGEMENT; GUIDELINES; AWARENESS AB Objective The purpose of this research is to assess short-term blood pressure change and hypertension incidence, and identify correlates of incident hypertension in the USA and Poland. Design and methods Population-based samples aged 45-64 years at enrollment from the Atherosclerosis Risk in Communities (ARIC) and the Pol-MONICA studies: including 3777 whites from Minneapolis, Minnesota, USA suburbs (urban), 3635 whites from Washington County, Maryland, USA (semi-rural) and 3109 blacks from Jackson, Mississippi, USA surveyed in 1987-1989 and 1990-1992; and 389 persons from Warsaw, Poland (urban) and 322 from Tarnobrzeg Province, Poland (semi-rural) surveyed in 1987-1988 and 1992-1993. Results Age-standardized systolic and diastolic blood pressures at both screens were 9-20 and 5-9 mmHg higher in the Polish samples than in US blacks, who had higher levels than US whites. Age-adjusted annual hypertension incidence in both Polish male cohorts (6-8%) was higher than that in US white men (4%) and approaching that of US black men (7%); rates were also higher in Polish female cohorts (8-9%) than in US black women (8%), but nearly twice those in US white women (4%). Factors independently related to hypertension incidence included age, family history, smoking, baseline blood pressures and body mass index, and increase in body mass index and alcohol consumption between screenings. After adjustment for these factors, annualized hypertension incidence was similar in US white and Polish men (2.3 and 2.7%) compared with US black men (3.4%), and in US white and Polish women (1.5 and 1.3%) compared with US black women (3.9%). Conclusions Despite substantial differences in blood pressure levels and age-standardized hypertension incidence rates, the differences in incidence between Polish and US white men appear to be explained largely by differences in risk factors for hypertension. J Hypertens 2000, 18:999-1006 (C) Lippincott Williams & Wilkins. C1 Univ N Carolina, Dept Biostat, Collaborat Studies Coordinating Ctr, Chapel Hill, NC 27514 USA. Stefan Cardinal Wyszynski Natl Inst Cardiol, Dept CVD Epidemiol & Prevent, Warsaw, Poland. Univ Alabama, Dept Biostat, Birmingham, AL 35294 USA. Jagiellonian Univ, Coll Med, Inst Publ Hlth, Div Clin Epidemiol, Krakow, Poland. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Univ Mississippi, Dept Prevent Med, Jackson, MS 39216 USA. RP Rywik, SL (reprint author), Univ N Carolina, Dept Biostat, Collaborat Studies Coordinating Ctr, CB8030,137 E Franklin Street, Chapel Hill, NC 27514 USA. FU NHLBI NIH HHS [N01-HV-08112, N01-HV-12243, N01-HV-59224] NR 42 TC 23 Z9 23 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD AUG PY 2000 VL 18 IS 8 BP 999 EP 1006 DI 10.1097/00004872-200018080-00002 PG 8 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 343FE UT WOS:000088690400002 PM 10953989 ER PT J AU Son, NH Murray, S Yanovski, J Hodes, RJ Weng, NP AF Son, NH Murray, S Yanovski, J Hodes, RJ Weng, NP TI Lineage-specific telomere shortening and unaltered capacity for telomerase expression in human T and B lymphocytes with age SO JOURNAL OF IMMUNOLOGY LA English DT Article ID REGULATED EXPRESSION; INCREASED APOPTOSIS; HUMAN FIBROBLASTS; CELLS; ACTIVATION; LENGTH; SUBSETS; CANCER; DIFFERENTIATION; MECHANISM AB Age effects on telomere length and telomerase expression in peripheral blood lymphocytes were analyzed from 121 normal individuals age newborn to 94 years and revealed several new findings. 1) Telomere shortening was observed in CD4(+) and CD8(+) T and B cells with age. However, the rate of telomere loss was significantly different in these populations, 35 +/- 8, 26 +/- 7, and 19 +/- 7 bp/year for CD4(+) and CD8(+) T and B cells, respectively, In addition, CD4(+) T cells had the longest average telomeres at all ages, followed by B cells, with CD8(+) T cell telomeres the shortest, suggesting that these lymphocyte populations may have different replicative histories in vivo. 2) Telomerase activity in freshly isolated T and B cells was indistinguishably low to undetectable at all ages but was markedly increased after Ag and costimulatory receptors mediated stimulation in vitro. Furthermore, age did not alter the magnitude of telomerase activity induced after stimulation of T or B lymphocytes through Ag and costimulatory receptors or in response tb PMA plus ionomycin treatment. 3) The levels of telomerase activity induced by in vitro stimulation varied among individual donors but were highly correlated with the outcome of telomere length change in CD4(+) T cells after Ag receptor-mediated activation. Together, these results indicate that rates of age-associated loss of telomere length in vivo in peripheral blood lymphocytes is specific to T and B cell subsets and that age does not significantly alter the capacity for telomerase induction in lymphocytes. C1 NIA, Immunol Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. NICHHD, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Weng, NP (reprint author), NIA, Immunol Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr,Box 21, Baltimore, MD 21224 USA. NR 44 TC 123 Z9 129 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 AUG 1 PY 2000 VL 165 IS 3 BP 1191 EP 1196 PG 6 WC Immunology SC Immunology GA 337CV UT WOS:000088340600004 PM 10903716 ER PT J AU Liang, H Reich, CF Pisetsky, DS Lipsky, PE AF Liang, H Reich, CF Pisetsky, DS Lipsky, PE TI The role of cell surface receptors in the activation of human B cells by phosphorothioate oligonucleotides SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HUMAN PERIPHERAL-BLOOD; LEUCINE METHYL-ESTER; BACTERIAL-DNA; CPG MOTIFS; LYMPHOCYTE-ACTIVATION; SCAVENGER RECEPTORS; IMMUNE STIMULATION; OLIGODEOXYNUCLEOTIDES; BINDING; LEUKOCYTES AB Phosphorothioate oligodeoxynucleotides (sODN) containing the CpG motif or TCG repeats induce T cell-independent polyclonal activation of human B cells. To elucidate the mechanism of this response, the role of cell surface receptors was investigated. Sepharose beads coated with stimulatory but not nonstimulatory sODNs induced B cell proliferation comparably with soluble sODNs. The B cell stimulatory activity of Sepharose-bound sODN did not result from free sODN released from the beads since media incubated with coated beads were inactive. Using FITC-labeled sODNs as probes, binding to human B cells could be detected by flow cytometry, Binding was rapid, saturable, initially temperature independent, but with a rapid off-rate. Competition studies indicated that both stimulatory sODNs and minimally stimulatory sODNs bound to the same receptor. By contrast, phosphodiester oligonucleotides with the same nucleotide sequence as sODNs and bacterial DNA inhibited the binding of sODNs to B cells minimally, Charge appeared to contribute to the binding of sODNs to B cells since binding of sODNs was competitively inhibited by negatively charged molecules, including fucoidan, poly I, and polyvinyl sulfate, These data indicate that human B cells bind sODNs by a receptor-mediated mechanism that is necessary but not sufficient for polyclonal activation. C1 Univ Texas, SW Med Ctr, Dept Internal Med, Harold C Simmons Arthrit Res Ctr, Dallas, TX 75235 USA. Duke Univ, Med Ctr, Dept Med,Durham Vet Affairs Med Ctr, Div Rheumatol Allergy & Clin Immunol, Durham, NC 27705 USA. RP Lipsky, PE (reprint author), NIH, 9000 Rockville Pike,Bldg 10,Room 9N228, Bethesda, MD 20892 USA. FU NIAID NIH HHS [2-PO1-AI31229-8] NR 33 TC 37 Z9 39 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 2000 VL 165 IS 3 BP 1438 EP 1445 PG 8 WC Immunology SC Immunology GA 337CV UT WOS:000088340600036 PM 10903748 ER PT J AU Tsuji, RF Kawikova, I Ramabhadran, R Akahira-Azuma, M Taub, D Hugli, TE Gerard, C Askenase, PW AF Tsuji, RF Kawikova, I Ramabhadran, R Akahira-Azuma, M Taub, D Hugli, TE Gerard, C Askenase, PW TI Early local generation of C5a initiates the elicitation of contact sensitivity by leading to early T cell recruitment SO JOURNAL OF IMMUNOLOGY LA English DT Article ID DELAYED-TYPE HYPERSENSITIVITY; SEROTONIN RELEASE; INTERFERON-GAMMA; ARTHUS REACTION; PROTEIN IP-10; P-SELECTIN; MAST-CELLS; IN-VIVO; EXPRESSION; COMPLEMENT AB We have shown previously that an early complement C5-dependent cascade is required to recruit T cells to elicit 24-h contact sensitivity (CS) responses. In this paper, we have characterized molecular events of this early required cascade by biochemically analyzing extracts of mouse ears undergoing elicitation of CS, Chemotactic activity was found after local Ag challenge, in CS ear extracts early (by 1 h), in CS ear extracts late (through 24 h), in previously immunized mice, but not in ears of vehicle-immunized or non-immune challenged mice. The early chemotactic activity at 2 h was likely caused by C5a, because it was neutralized in vitro. by anti-C5a Ab, was inactive on C5aR-deficient (C5aR(-/-)) macrophages, and was absent In C5-deficient mice. The activity was present in T cell-deficient mice, but elaboration was Ag-specific. This T cell-independent, Ag specific elaboration of C5a early in CS ear responses likely led to T cell recruitment, because subsequent local IFN-gamma mRNA and protein expression, as markers of T cell arrival and activation, began by 4 h after Ag challenge. In contrast to early C5a chemotactic activity, late chemotactic activity 24 h after Ag challenge was unaffected by anti-C5, was active on C5aR(-/-) macrophages, was T cell-dependent, and by ELISA appeared largely due to chemokines (macrophage-inflammatory protein-1 alpha and 1 beta, IFN-gamma-inducible protein-10, and monocyte chemoattractant protein-1). Importantly, early generation of C5a was required for T cell recruitment because C5aR(-/-) mice had absent 24-h CS, Taken together, these findings indicate an important linkage of C5a as a component of early activated innate immunity that is required for later elicitation of acquired T cell immunity, probably by facilitating the initial recruitment of T cells into the Ag-challenged local site in CS responses. C1 Noda Inst Sci Res, Noda, Chiba 2780037, Japan. Yale Univ, Sch Med, Dept Internal Med, Allergy & Clin Immunol Sect, New Haven, CT 06520 USA. NIA, Clin Immunol Sect, NIH, Geriatr Res Ctr, Baltimore, MD 21224 USA. Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA. Harvard Univ, Sch Med, Childrens Hosp, Perlmutter Lab, Boston, MA 02115 USA. RP Tsuji, RF (reprint author), Noda Inst Sci Res, 399 Noda, Noda, Chiba 2780037, Japan. FU NHLBI NIH HHS [P50 HL-56389]; NIAID NIH HHS [AI-43371] NR 47 TC 96 Z9 99 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 2000 VL 165 IS 3 BP 1588 EP 1598 PG 11 WC Immunology SC Immunology GA 337CV UT WOS:000088340600056 PM 10903768 ER PT J AU Dybul, M Mercier, G Belson, M Hallahan, CW Liu, SY Perry, C Herpin, B Ehler, L Davey, RT Metcalf, JA Mican, JM Seder, RA Fauci, AS AF Dybul, M Mercier, G Belson, M Hallahan, CW Liu, SY Perry, C Herpin, B Ehler, L Davey, RT Metcalf, JA Mican, JM Seder, RA Fauci, AS TI CD40 ligand trimer and IL-12 enhance peripheral blood mononuclear cells and CD4(+) T cell proliferation and production of IFN-gamma in response to p24 antigen in HIV-infected individuals: Potential contribution of anergy to HIV-specific unresponsiveness SO JOURNAL OF IMMUNOLOGY LA English DT Article ID ACTIVE ANTIRETROVIRAL THERAPY; HUMAN DENDRITIC CELLS; IN-VITRO; ANTICYTOMEGALOVIRUS THERAPY; INTERLEUKIN-2 PRODUCTION; IMMUNE-RESPONSES; INTERFERON-GAMMA; CO-STIMULATION; RESTORATION; AIDS AB It has been suggested that CD4(+) T cell proliferative responses to HIV p24 Ag may be important in the control of HIV infection. However, these responses are minimal or absent in many EW-infected individuals, Furthermore, while in vitro and in vivo responses to non-HIV recall Ags improve upon administration of highly active antiretroviral therapy, there does not appear to be a commensurate enhancement of HIV-specific immune responses. It is possible that CD4(+) p24-specific T cells are deleted early in the course of infection. However, it is also possible that a discrete unresponsiveness, or anergy, contributes to the lack of proliferation to p24. To evaluate the possible contribution of unresponsiveness to the lack of CD4(+) T cell proliferation to p24 in HIV-infected individuals, we attempted to overcome unresponsiveness. CD40 ligand trimer (CD40LT) and IL-12 significantly increased PBMC and CD4(+) T cell proliferative responses to p24 Ag in HIV-infected, but not uninfected, individuals. No increase in proliferative response to CMV Ag was observed. CDL40LT exerted its effect through B7-CD28-dependent and IL-12- and IL-15-independent mechanisms. Finally, the increase in proliferation with CD40LT and IL-12 was associated with an augmented production of IFN-gamma in most, but not all, individuals. These data suggest the possible contribution of HIV-specific unresponsiveness to the lack of CD4(+) T cell proliferation to p24 Ag in HIV-infected individuals and that clonal deletion alone does not explain this phenomenon. They also indicate the potential for CD40LT and IL-12 as immune-based therapies for HIV infection. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. NIH, Warren Magneson Clin Res Ctr, Bethesda, MD 20892 USA. RP Dybul, M (reprint author), NIAID, Immunoregulat Lab, NIH, Bldg 10,Room 11N204, Bethesda, MD 20892 USA. NR 54 TC 40 Z9 40 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 2000 VL 165 IS 3 BP 1685 EP 1691 PG 7 WC Immunology SC Immunology GA 337CV UT WOS:000088340600068 PM 10903780 ER PT J AU Emery, S Capra, WB Cooper, DA Mitsuyasu, RT Kovacs, JA Vig, P Smolskis, M Saravolatz, LD Lane, HC Fyfe, GA Curtin, PT AF Emery, S Capra, WB Cooper, DA Mitsuyasu, RT Kovacs, JA Vig, P Smolskis, M Saravolatz, LD Lane, HC Fyfe, GA Curtin, PT CA Int Interleukin-2 Study Grp TI Pooled analysis of 3 randomized, controlled trials of interleukin-2 therapy in adult human immunodeficiency virus type 1 disease SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 5th Conference on Retroviruses and Opportunistic Infections CY FEB 01-05, 1998 CL CHICAGO, ILLINOIS ID DOSE SUBCUTANEOUS INTERLEUKIN-2; ACTIVE ANTIRETROVIRAL THERAPY; CD4 CELL COUNTS; T-CELLS; INTRAVENOUS INTERLEUKIN-2; INFECTED PATIENTS; CUBIC MILLIMETER; CLINICAL-TRIALS; HIV-1 INFECTION; METAANALYSIS AB We collected human immunodeficiency virus (HIV) disease progression, survival, most recent CD4 cell count, and plasma HIV RNA levels from patients (n = 157) who participated in randomized clinical trials of interleukin (IL)-2 that commenced before 1995. Data were available for 155 (99%) patients. Statistical analyses were based on the intention-to-treat principle. Median follow-up was 28 months and 30 months for control and IL-2 patients, respectively. Twenty-five (16%) patients developed AIDS or died during follow-up (16 control patients vs. 9 IL-2 patients; R-2 = 0.57; P = .22). Mean change from baseline CD4 cell count was significantly higher in patients randomized to receive IL-2 (368 vs. 153 cells/mu L; P = .003). Mean change from baseline plasma HIV RNA was significantly lower in patients randomized to receive IL-2 (-0.98 vs. -0.63 log copies/mL; P = .004). Significant improvements in CD4 cell count and plasma HIV RNA in recipients of IL-2 relative to control patients were associated with a nonsignificant trend toward improved clinical outcome. C1 Univ New S Wales, Natl Ctr HIV Epidemiol & Clin Res, Sydney, NSW 2010, Australia. Chiron Corp, Emeryville, CA 94608 USA. Univ Calif Los Angeles, CARE Ctr, Los Angeles, CA USA. NIH, Ctr Clin, Bethesda, MD 20892 USA. NIAID, NIH, Bethesda, MD 20892 USA. Henry Ford Hosp, Detroit, MI 48202 USA. RP Emery, S (reprint author), Univ New S Wales, Natl Ctr HIV Epidemiol & Clin Res, 376 Victoria St, Sydney, NSW 2010, Australia. RI Emery, Sean/H-4920-2013 OI Emery, Sean/0000-0001-6072-8309 NR 44 TC 80 Z9 81 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD AUG PY 2000 VL 182 IS 2 BP 428 EP 434 DI 10.1086/315736 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 347WJ UT WOS:000088951900007 PM 10915072 ER PT J AU Fogelman, I Davey, V Ochs, HD Elashoff, M Feinberg, MB Mican, J Siegel, JP Sneller, M Lane, HC AF Fogelman, I Davey, V Ochs, HD Elashoff, M Feinberg, MB Mican, J Siegel, JP Sneller, M Lane, HC TI Evaluation of CD4(+) T cell function in vivo in HIV-infected patients as measured by bacteriophage phiX174 immunization SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 12th World AIDS Conference CY JUN 28-JUL 03, 1998 CL GENEVA, SWITZERLAND ID ACTIVE ANTIRETROVIRAL THERAPY; DEFECTIVE HUMORAL IMMUNITY; ANTIBODY-RESPONSES; CD40 LIGAND; HYPER-IGM; IMMUNODEFICIENCY; DEFICIENCY; EXPRESSION; DISEASE; ABNORMALITIES AB Bacteriophage phiX174 immunization was used to measure CD4(+) T cell function in vivo in human immunodeficiency virus (HIV)-infected patients across all disease stages. Function was evaluated by measuring the ability of T cells to provide help to B cells in antibody production, amplification, and isotype switching. A total of 33 patients and 10 controls received 3 bacteriophage phiX174 immunizations 6 weeks apart. The patients' responses regarding bacteriophage-specific total antibody titers and IgG titers were quantitatively and qualitatively inferior to the controls' responses. Overall, 7 of 33 patients had normal T cell function. Baseline CD4 counts provided the strongest correlation with total antibody and IgG titers. HIV RNA had a weaker association with responses but had some predictive power among patients with a CD4 count >200 cells/mu L. Bacteriophage phiX174 immunization seems to be a useful tool for measuring immune function in vivo, which suggests that most HIV-infected patients may have abnormal CD4(+) T cell function despite adequate antiretroviral treatment. C1 US FDA, Ctr Biol Evaluat & Res, Rockville, MD 20857 USA. US FDA, Ctr Drug Evaluat & Res, Rockville, MD 20857 USA. NIAID, NIH, Bethesda, MD 20892 USA. Univ Washington, Sch Med, Dept Pediat, Seattle, WA 98195 USA. Emory Univ, Vaccine Res Ctr, Atlanta, GA 30322 USA. RP Fogelman, I (reprint author), 3962 Germantown Rd, Edgewater, MD 21037 USA. NR 24 TC 39 Z9 40 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD AUG PY 2000 VL 182 IS 2 BP 435 EP 441 DI 10.1086/315739 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 347WJ UT WOS:000088951900008 PM 10915073 ER PT J AU Zhou, JF Dong, YH Zhao, XL Lee, SW Amin, A Ramaswamy, S Domagala, A Musser, JM Drlica, K AF Zhou, JF Dong, YH Zhao, XL Lee, SW Amin, A Ramaswamy, S Domagala, A Musser, JM Drlica, K TI Selection of antibiotic-resistant bacterial mutants: Allelic diversity among fluoroquinolone-resistant mutations SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID NEW-YORK-CITY; MYCOBACTERIUM-TUBERCULOSIS; DNA GYRASE; ESCHERICHIA-COLI; STREPTOCOCCUS-PNEUMONIAE; QUINOLONE RESISTANCE; TOPOISOMERASE-IV; STAPHYLOCOCCUS-AUREUS; ANTIMICROBIAL AGENTS; SOS RESPONSE AB To obtain a general framework for understanding selection of antibiotic-resistant mutants, allelic diversity was examined with about 600 fluoroquinolone-resistant mutants of mycobacteria. Selection at low fluoroquinolone concentration produced many low-level resistance mutants. Some of these contained mutations that conferred unselected antibiotic resistance; none contained alterations in the quinolone-resistance-determining region of the GyrA protein, the principal drug target. As selection pressure increased, a variety of GyrA variants became prevalent. High concentrations of antibiotic reduced the variety to a few types, and eventually a concentration was reached at which no mutant was recovered. That concentration defined a threshold for preventing the selection of resistance. The pattern of variants selected, which was also strongly influenced by antibiotic structure, readily explained the variants present in clinical isolates. Thus, resistance arises from selection of mutants whose identity depends on drug concentration and structure, both of which can be manipulated to restrict selection. C1 Publ Hlth Res Inst, New York, NY 10016 USA. Baylor Coll Med, Dept Pathol, Inst Human Bacterial Pathogenesis, Houston, TX 77030 USA. NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. Warner Lambert Parke Davis, Parke Davis Pharmaceut Res Div, Ann Arbor, MI USA. RP Publ Hlth Res Inst, 455 1st Ave, New York, NY 10016 USA. EM drlica@phri.nyu.edu FU NIAID NIH HHS [AI35257, AI37004] NR 37 TC 95 Z9 99 U1 1 U2 3 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0022-1899 EI 1537-6613 J9 J INFECT DIS JI J. Infect. Dis. PD AUG PY 2000 VL 182 IS 2 BP 517 EP 525 DI 10.1086/315708 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 347WJ UT WOS:000088951900018 PM 10915083 ER PT J AU Nathanson, N Mathieson, BJ AF Nathanson, N Mathieson, BJ TI Biological considerations in the development of a human immunodeficiency virus vaccine SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 37th Annual Meeting of the Infectious-Diseases-Society-of-America CY NOV 18-21, 1999 CL PHILADELPHIA, PENNSYLVANIA SP Infect Dis Soc Amer ID CYTOTOXIC-T-CELL; ACTIVE ANTIRETROVIRAL THERAPY; LIVE ATTENUATED SIV; BLOOD-BANK COHORT; RHESUS MACAQUES; IMMUNE-RESPONSES; HIV TYPE-1; VAGINAL CHALLENGE; AIDS VACCINE; NEUTRALIZING ANTIBODIES AB Over the last 12 years, many human immunodeficiency virus (HIV) vaccine candidates have been tried in humans, with disappointing results. In particular, recombinant envelope proteins have failed to elicit strong cellular immune responses or neutralizing antibody against many wild-type isolates of HIV-1, Attenuated strains of simian immunodeficiency virus (SIV), although capable of protecting against virulent strains of SIV, often retain residual pathogenicity. These difficulties suggest that it will be necessary to address a number of biological questions that underpin the rational development of an AIDS vaccine: (1) Will natural infection with HIV protect against superinfection? (2) Is partial protection induced by an HIV vaccine adequate to prevent AIDS? (3) What are the immune correlates of protection for an AIDS vaccine? (4) Will a monotypic HIV-1 vaccine confer cross-clade immunity? (5) Is mucosal immunity important for an effective AIDS vaccine? (6) Is there a rationale for therapeutic immunization! Ongoing research that is addressing these questions should lead to the formulation of a safe and effective AIDS vaccine. C1 NIH, Off AIDS Res, Bethesda, MD 20892 USA. RP Nathanson, N (reprint author), NIH, Off AIDS Res, Bldg 2,Rm 4W04,2 Ctr Dr, Bethesda, MD 20892 USA. NR 101 TC 34 Z9 34 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD AUG PY 2000 VL 182 IS 2 BP 579 EP 589 DI 10.1086/315707 PG 11 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 347WJ UT WOS:000088951900027 PM 10915092 ER PT J AU Goedert, JJ Hatzakis, A Maloney, EM Eyster, ME AF Goedert, JJ Hatzakis, A Maloney, EM Eyster, ME CA Multicenter Hemophilia Cohort Stud TI Increased liver decompensation risk with atypical hepatitis C virus antibody levels SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 10th International Symposium on Viral Hepatitis and Liver Disease CY APR, 2000 CL ATLANTA, GEORGIA ID HUMAN-IMMUNODEFICIENCY-VIRUS; HEMOPHILIC MEN; INFECTION; DISEASE; PROGRESSION; COHORT; CELLS; HIV AB Knowledge of serum markers of Liver decompensation would facilitate care of patients with hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections. HCV load and anti-c33c and anti-NS5 levels did not distinguish 28 HCV- and HIV-positive predecompensation patients from 28 matched control patients, whereas more patients than controls had high anti-c100(p) and low anti-c22(p). In multivariate analysis, decompensation was associated with high anti-c100(p) titer (greater than or equal to 1:4050; odds ratio [OR], 3.4; 95% confidence interval [CI], 1.1-11.5) and low anti-c22(p) (<1:36,450; OR, 3.0; 95% CI, 1.0-10.2) and with antibody band strength at 1:50 dilution (anti-c100[p] OR, 7.0; 95% CI, 1.7-48.9; anti-c22[p] OR, 7.1; 95% CI, 1.7-49.2), With high anti-c100(p) or low anti-c22(p), sensitivity for decompensation was 86%-96% and specificity was 21%-36%; with bath markers, sensitivity was 29%-32% and specificity was 93%-96%. Although the mechanisms for these associations are unknown, if these findings are verified in other populations, anti-c100(p) and anti-c22(p) might be valuable surrogate markers for liver decompensation risk. C1 NCI, Viral Epidemiol Branch, Rockville, MD USA. Univ Athens, Dept Hyg & Epidemiol, Natl Retrovirus Reference Ctr, Athens, Greece. Penn State Univ, Coll Med, Dept Med, Div Hematol Oncol, Hershey, PA USA. RP Goedert, JJ (reprint author), 6120 Execut Blvd,Ste 8012,MSC 7248, Rockville, MD 20852 USA. FU NCI NIH HHS [N01-CP33002] NR 15 TC 6 Z9 6 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD AUG PY 2000 VL 182 IS 2 BP 590 EP 594 DI 10.1086/315712 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 347WJ UT WOS:000088951900028 PM 10915093 ER PT J AU Al-Harthi, L Wright, DJ Anderson, D Cohen, M Matityahu, D Cohn, J Cu-Unvin, S Burns, D Reichelderfer, P Lewis, S Beckner, S Kovacs, A Landay, A AF Al-Harthi, L Wright, DJ Anderson, D Cohen, M Matityahu, D Cohn, J Cu-Unvin, S Burns, D Reichelderfer, P Lewis, S Beckner, S Kovacs, A Landay, A TI The impact of the ovulatory cycle on cytokine production: Evaluation of systemic, cervicovaginal, and salivary compartments SO JOURNAL OF INTERFERON AND CYTOKINE RESEARCH LA English DT Article ID MENSTRUAL-CYCLE; SECRETORY COMPONENT; MONONUCLEAR-CELLS; FOLLICULAR-FLUID; GONADAL-STEROIDS; EPITHELIAL-CELLS; BLOOD-LEVELS; INTERLEUKIN-6; EXPRESSION; WOMEN AB To understand the impact of the menstrual cycle on immunologic parameters, we measured the level of cytokines and chemokines from plasma, cervicovaginal lavage (CVL), and saliva samples of 6 premenopausal women during the follicular and luteal phases of the ovulatory cycle. We demonstrate that the level of plasma interleukin-8 (IL-8) was 4-fold higher during the follicular phase than the luteal phase (p = 0.004), whereas plasma IL-1 beta, IL-4, IL-6, IL-10, interferon-gamma (IFN-gamma), transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), macrophage inflammatory protein-1 alpha (MIP-1 alpha), and TNF receptor II (TNFR II) were not altered during the ovulatory cycle. In the vaginal compartment, as measured from CVL samples, the levels of IL-6 and IL-1 beta were both 5-fold higher in the follicular than the luteal phase (p = 0.0002 and 0.03, respectively). Salivary cytokine and chemokine samples were similar when measured during the luteal and the follicular phases. Additional analysis of lymphocyte subsets for phenotypic and functional markers indicated that they were not influenced by the ovulatory cycle. Collectively, these data suggest that IL-6, IL-8, and IL-1 beta are differentially regulated during the ovulatory cycle. C1 Rush Presbyterian St Lukes Med Ctr, Dept Immunol Microbiol, Chicago, IL 60612 USA. WESTAT Corp, Rockville, MD 20850 USA. Harvard Univ, Sch Med, Boston, MA 02115 USA. Cook Cty Hosp, Chicago, IL 60612 USA. SUNY Hlth Sci Ctr, Brooklyn, NY 11203 USA. Wayne State Univ, Detroit, MI 48201 USA. Brown Univ, Providence, RI 02906 USA. NICHHD, NIH, Bethesda, MD 20892 USA. LAC & USC Maternal & Child HIV Management & Res C, Los Angeles, CA 90033 USA. RP Al-Harthi, L (reprint author), Rush Presbyterian St Lukes Med Ctr, Dept Immunol Microbiol, 1653 W Congress Pkwy,Room 1577 JSC, Chicago, IL 60612 USA. FU NICHD NIH HHS [N01-HD-3-3162] NR 25 TC 50 Z9 53 U1 0 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1079-9907 J9 J INTERF CYTOK RES JI J. Interferon Cytokine Res. PD AUG PY 2000 VL 20 IS 8 BP 719 EP 724 DI 10.1089/10799900050116426 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Immunology SC Biochemistry & Molecular Biology; Cell Biology; Immunology GA 345FZ UT WOS:000088807100006 PM 10954915 ER PT J AU Dallinger, G Puttaraju, M Mitchell, LG Yancey, KB Hintner, H Bauer, JW AF Dallinger, G Puttaraju, M Mitchell, LG Yancey, KB Hintner, H Bauer, JW TI Collagen 17A1 gene correction using spliceosome mediated RNA trans-splicing (SMaRT (TM)) technology SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Meeting Abstract C1 Gen Hosp, Dept Dermatol, Salzburg, Austria. INTRONN LLC, Durham, NC USA. NIH, Dermatol Branch, Bethesda, MD 20892 USA. NR 0 TC 9 Z9 9 U1 0 U2 0 PU BLACKWELL SCIENCE 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 2000 VL 115 IS 2 MA HB2 BP 332 EP 332 DI 10.1046/j.1523-1747.2000.115002332.x PG 1 WC Dermatology SC Dermatology GA 345FK UT WOS:000088805800034 ER PT J AU Fitzhugh, DJ Naik, S Gonzalez, E Caughman, SW Hwang, ST AF Fitzhugh, DJ Naik, S Gonzalez, E Caughman, SW Hwang, ST TI CC chemokine receptor 6 (CCR6) is a marker for memory T cells that arrest on activated human dermal microvascular endothelium under shear stress SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Meeting Abstract C1 NCI, Dermatol Branch, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Dept Dermatol, Atlanta, GA 30322 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL SCIENCE 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 2000 VL 115 IS 2 MA HB7 BP 333 EP 333 PG 1 WC Dermatology SC Dermatology GA 345FK UT WOS:000088805800037 ER PT J AU Gerelsaikhan, T Turner, RJ AF Gerelsaikhan, T Turner, RJ TI Membrane topology and function of the secretory Na+-K+-2Cl(-) cotransporter (NKCC1) SO JOURNAL OF KOREAN MEDICAL SCIENCE LA English DT Article; Proceedings Paper CT International Symposium on Exocrine Glands CY APR 26-27, 2000 CL SEOUL, SOUTH KOREA DE cation-chloride cotransporter; fluid secretion; membrane spanning region; transmembrane topology ID K-CL COTRANSPORTER; MOLECULAR CHARACTERIZATION; BUMETANIDE BINDING; PROTEIN; PHOSPHORYLATION; TRANSPORT; DOMAINS; ION C1 Natl Inst Dent & Craniofacial Res, Membrane Biol Sect, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Turner, RJ (reprint author), Natl Inst Dent & Craniofacial Res, Membrane Biol Sect, Gene Therapy & Therapeut Branch, NIH, Bldg 10,Rm 1A01, Bethesda, MD 20892 USA. NR 14 TC 2 Z9 2 U1 0 U2 1 PU KOREAN ACAD MEDICAL SCIENCES PI SEOUL PA 302 75 DONG DU ICHON, DONG YONGSAN KU, SEOUL 140 031, SOUTH KOREA SN 1011-8934 J9 J KOREAN MED SCI JI J. Korean Med. Sci. PD AUG PY 2000 VL 15 SU S BP S3 EP S4 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 349NF UT WOS:000089050600002 PM 10981494 ER PT J AU Hsin, LW Webster, EL Chrousos, GP Gold, PW Eckelman, WC Contoreggi, C Rice, KC AF Hsin, LW Webster, EL Chrousos, GP Gold, PW Eckelman, WC Contoreggi, C Rice, KC TI Synthesis of [H-3] (4-fluorobutyl)propyl[2,5,6-trimethyl-7-(2,4,6-trimethylphenyl)pyrrolo[2 ,3-d]pyrimidin-4-yl]amine: A potent radioligand for corticotropin-releasing hormone type 1 receptor SO JOURNAL OF LABELLED COMPOUNDS & RADIOPHARMACEUTICALS LA English DT Article DE corticotropin-releasing hormone; radioligand; tritium; antalarmin; positron emission tomography ID PITUITARY AB [H-3](4-Fluorobutyl)propyl[2,5,6-trimethyl-7-(2,4,6-trimethylphenyl)pyrrolo[2,3-d]pyrimidin-4-yl]amine ([H-3]LWH-154), a novel potent radiolabelled analog of the nonpeptide corticotropin-releasing hormone type I receptor (CRHR1) selective antagonist, butylethyl[2,5,6-trimethyl-7-(2,4,6-trimethylphenyl)pyrrolo[2,3-d]pyrimidin-4-yl]amine (antalarmin), was prepared for the development of positron emission tomography radiotracers for CRHR1 and evaluation as a nonpeptide radioligand for use in pharmacological studies. The precursor (4-fluorobutyl)prop-2-enyl[2,5,6-trimethyl-7-(2,4,6-trimethylphenyl)pyrrolo[2,3-d]pyrimidin-4-yl]amine (6) for tritiation was prepared in two steps from 3 in 76% total yield. Catalytic reduction of unsaturated fluoride 6 using tritium gas and palladium as catalyst gave [H-3]LWH-154. After HPLC purification, [H-3]LWH-154 of high radiochemical purity was obtained with a specific activity of 69 Ci/mmol. C1 NIDDK, Med Chem Lab, Bethesda, MD 20892 USA. NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. NICHD, Pediat Endocrinol Sect, PREB, Bethesda, MD 20892 USA. NIH, Dept Nucl Med, Ctr Clin, Bethesda, MD 20892 USA. NIDA, Mol Neurobiol Sect, Baltimore, MD 21224 USA. RP Hsin, LW (reprint author), NIDDK, Med Chem Lab, Bethesda, MD 20892 USA. OI HSIN, LING-WEI/0000-0001-5018-4491 NR 14 TC 11 Z9 11 U1 0 U2 0 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0362-4803 J9 J LABELLED COMPD RAD JI J. Label. Compd. Radiopharm. PD AUG PY 2000 VL 43 IS 9 BP 899 EP 908 DI 10.1002/1099-1344(200008)43:9<899::AID-JLCR375>3.0.CO;2-N PG 10 WC Biochemical Research Methods; Chemistry, Medicinal; Chemistry, Analytical SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Chemistry GA 338YB UT WOS:000088449900006 ER PT J AU Gorelick, RJ Lifson, JD Yovandich, JL Rossio, JL Piatak, M Scarzello, AJ Knott, WB Bess, JW Fisher, BA Flynn, BM Henderson, LE Arthur, LO Benveniste, RE AF Gorelick, RJ Lifson, JD Yovandich, JL Rossio, JL Piatak, M Scarzello, AJ Knott, WB Bess, JW Fisher, BA Flynn, BM Henderson, LE Arthur, LO Benveniste, RE TI Mucosal challenge of Macaca nemestrina with simian immunodeficiency virus (SIV) following SIV nucleocapsid mutant DNA vaccination SO JOURNAL OF MEDICAL PRIMATOLOGY LA English DT Article DE AIDS; intrarectal inoculation; pig-tailed macaques; SIV Mne clone E11S; vaccines ID LONG-TERM PROTECTION; ENVELOPE GLYCOPROTEIN; VIRAL REPLICATION; IN-VITRO; MACAQUES; INFECTION; RNA; VACCINES; IMMUNIZATION; LENTIVIRUS AB A simian immunodeficiency virus (SIV)(Mne) DNA clone was constructed that produces viruses containing a four amino acid deletion in the second zinc finger of the nucleocapsid (NC) domain of the Gag polyprotein. Viruses produced from this clone, although noninfectious both in vitro and in vivo, complete a majority of the steps in a single retroviral infection cycle. Eight pig-tailed macaques (Macaca nemestrina) were inoculated intramuscularly and subcutaneously three times over the course of 24 weeks with the NC mutant expressing DNA. These macaques, and four controls, were then challenged mucosally (intrarectally) with the homologous virus (SIV Mne CL E11S) and monitored for evidence of infection and clinical disease. Prior to challenge, a measurable humoral immune response was noted in four of eight immunized macaques. After challenge, all 12 macaques became infected, although four immunized animals greatly restricted their viral replication, and one immunized animal that controlled replication remains antibody negative. No disease has been evidence during the 46-week period of monitoring after challenge. C1 NCI, AIDS Vaccine Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, Basic Res Labs, Frederick, MD 21702 USA. NCI, Intramural Res Support Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, Anim Sci Branch, Bethesda, MD 20892 USA. RP Arthur, LO (reprint author), NCI, AIDS Vaccine Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RI Bess, Jr., Julian/B-5343-2012 FU NCI NIH HHS [N01-CO-56000] NR 28 TC 5 Z9 5 U1 0 U2 1 PU MUNKSGAARD INT PUBL LTD 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 2000 VL 29 IS 3-4 BP 209 EP 219 DI 10.1034/j.1600-0684.2000.290314.x PG 11 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 359GF UT WOS:000089602600014 PM 11085583 ER PT J AU Teubner, B Degen, J Sohl, G Guldenagel, M Bukauskas, FF Trexler, EB Verselis, VK De Zeeuw, CI Lee, CG Kozak, CA Petrasch-Parwez, E Dermietzel, R Willecke, K AF Teubner, B Degen, J Sohl, G Guldenagel, M Bukauskas, FF Trexler, EB Verselis, VK De Zeeuw, CI Lee, CG Kozak, CA Petrasch-Parwez, E Dermietzel, R Willecke, K TI Functional expression of the murine connexin 36 gene coding for a neuron-specific gap junctional protein SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE gap junctions; electrical synapses; neuronal connexin; transcriptional start site; Cx36 ID MOLECULAR-CLONING; RAT HIPPOCAMPUS; BRAIN; MOUSE; RETINA; OLIGODENDROCYTES; LOCALIZATION; INTERNEURONS; CONDUCTANCE; ASTROCYTES AB The mouse connexin 36 (Cx36) gene was mapped on chromosome 2 and an identical transcriptional start site was determined in brain and retina on exon I. Rabbit polyclonal antibodies to the presumptive cytoplasmic loop of the Cx36 protein recognized in immunohistochemical analyses Cx36 expression in the retina, olfactory bulb, hippocampus, inferior olive and cerebellum. In olivary neurons strong punctate labeling at dendritic cell contacts and weaker labeling in the cytoplasm of dendrites were shown by immune electron microscopy. After expression of mouse Cx36 cDNA in human HeLa cells, neurobiotin transfer was increased 1.8-fold and electrical conductance at least 15-fold compared to untransfected HeLa cells. No Lucifer Yellow transfer was detected in either untransfected or Cx36 transfected HeLa cells. Single Cx36 channels ill transfected HeLa cells showed a unitary conductance of 14.3 +/- 0.8 pS. The sensitivity of Cx36 channels to transjunctional voltage was low in both HeLa-Cx36 cells and Xenopus oocytes expressing mouse Cx36. No increased transfer of neurobiotin was detected in heterotypic gap junctions formed by Cx36 and 9 other connexins expressed in HeLa cells. Our results suggest that Cx36 channels function as electrical synapses for transmission of electrical and metabolic signals between neurons in the central nervous system. C1 Univ Bonn, Inst Genet, Abt Mol Genet, D-53117 Bonn, Germany. Albert Einstein Coll Med, Dept Neurosci, New York, NY USA. Erasmus Univ, Dept Anat, Rotterdam, Netherlands. NIAID, NIH, Bethesda, MD 20892 USA. Ruhr Univ Bochum, Inst Anat, Bochum, Germany. RP Willecke, K (reprint author), Univ Bonn, Inst Genet, Abt Mol Genet, Romerstr 164, D-53117 Bonn, Germany. FU NINDS NIH HHS [R01 NS036706, NS367060] NR 41 TC 154 Z9 159 U1 1 U2 2 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 2000 VL 176 IS 3 BP 249 EP 262 DI 10.1007/s002320001094 PG 14 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 342RN UT WOS:000088659100006 PM 10931976 ER PT J AU Yu, ZF Zhou, DH Cheng, GJ Mattson, MP AF Yu, ZF Zhou, DH Cheng, GJ Mattson, MP TI Neuroprotective role for the p50 subunit of NF-kappa B in an experimental model of Huntington's disease SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE apoptosis; calcium; caspase; mitochondrial toxin; nitropropionic acid; striatal neurons; oxidative stress; transcription ID MANGANESE SUPEROXIDE-DISMUTASE; NERVE GROWTH-FACTOR; FOCAL CEREBRAL-ISCHEMIA; NECROSIS-FACTOR-ALPHA; CAG REPEAT LENGTH; 3-NITROPROPIONIC ACID; INDUCED APOPTOSIS; HIPPOCAMPAL-NEURONS; CALCIUM HOMEOSTASIS; OXIDATIVE STRESS AB Prototypical NF-kappaB consists of a transcription factor dimer of p50 and p65, and an inhibitory subunit called I-kappaB. NF-kappaB is activated in neurons in response to excitotoxic, metabolic, and oxidative stress. Cell-culture data suggest that activation of NF-kappaB can prevent neuronal apoptosis, but its role in vivo is unclear and the specific kappaB subunits involved are unknown. In Huntington's disease (HD), striatal neurons degenerate, and a similar pattern of neuronal vulnerability occurs in rats and mice following exposure to the mitochondrial toxin 3-nitropropionic acid (3NP). We report that mice lacking the p50 subunit of NF-kappaB exhibit increased damage to striatal neurons following administration of 3NP. The neuronal death occurs by apoptosis as indicated by increased caspase activation and DNA fragmentation into oligonucleosomes. NF-kappaB activity is markedly increased in striatum 24-72 h following 3NP administration in wild-type mice, but not in mice lacking p50, indicating that p50 is necessary for the vast majority of 3NP-induced NF-kappaB DNA-binding activity in striatum. Cultured striatal neurons from p50-/- mice exhibited enhanced oxidative stress, perturbed calcium regulation, and increased cell death following exposure to 3NP, suggesting a direct adverse effect of p50 deficiency in striatal neurons. C1 NIA, Sanders Brown Res Ctr Aging, Gerontol Res Ctr, Baltimore, MD 21224 USA. NIA, Div Rheumatol Allergy & Immunol, Gerontol Res Ctr, Baltimore, MD 21224 USA. NIA, Dept Internal Med, Gerontol Res Ctr, Baltimore, MD 21224 USA. NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Mattson, MP (reprint author), NIA, Sanders Brown Res Ctr Aging, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 67 TC 58 Z9 60 U1 0 U2 2 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 PY 2000 VL 15 IS 1 BP 31 EP 44 DI 10.1385/JMN:15:1:31 PG 14 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 395UC UT WOS:000166598800004 PM 11211235 ER PT J AU Yamashita, RA Sellers, JR Anderson, JB AF Yamashita, RA Sellers, JR Anderson, JB TI Identification and analysis of the myosin superfamily in Drosophila: a database approach SO JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY LA English DT Review ID REGULATORY LIGHT-CHAIN; ACTIN-BASED MOTOR; UNCONVENTIONAL MYOSIN; NONMUSCLE MYOSIN; HEAVY-CHAIN; PHOSPHORYLATION SITES; IMAGINAL DISC; NINAC KINASE; EGG CHAMBER; SALT-BRIDGE AB The recent sequencing of the genome of Drosophila melanogaster has provided a valuable resource for mining the database for genes of interest. We took advantage of this opportunity in an attempt to identify novel myosins in Drosophila and confirm the presence of the previously identified myosins from classes I, II, III, V, VI, and VII. The Drosophila database annotators predicted the structure of three additional proteins which we identified as novel unconventional myosins, two of which fell into classes XV and XVIII, respectively. Our own efforts predicted the presence of four additional partial sequences that appear to be myosin proteins which did not fall into any specific class. In the future comparative genomics will hopefully lead to the placement of these myosins into new classes. C1 NHLBI, Mol Cardiol Lab, NIH, Bethesda, MD 20892 USA. RP Sellers, JR (reprint author), NHLBI, Mol Cardiol Lab, NIH, Bethesda, MD 20892 USA. NR 74 TC 39 Z9 39 U1 0 U2 4 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0142-4319 J9 J MUSCLE RES CELL M JI J. Muscle Res. Cell Motil. PD AUG PY 2000 VL 21 IS 6 BP 491 EP 505 DI 10.1023/A:1026589626422 PG 15 WC Cell Biology SC Cell Biology GA 379GX UT WOS:000165634600001 PM 11206129 ER PT J AU Kibayashi, C Aoyagi, S Wang, TC Saito, K Daly, JW Spande, TF AF Kibayashi, C Aoyagi, S Wang, TC Saito, K Daly, JW Spande, TF TI Determination of absolute stereochemistry and an alternative synthesis of homopumiliotoxin 223G: Identification on chiral GC columns with the natural alkaloid SO JOURNAL OF NATURAL PRODUCTS LA English DT Article ID (+)-ALLOPUMILIOTOXIN-339A; PYRROLIZIDINES; QUINOLIZIDINES; PUMILIOTOXINS; INDOLIZIDINES AB An alternative asymmetric synthesis of (+)-(1S,9aS)-homopumiliotoxin 223G (1) was accomplished via (1R,2R,9aS)-1-(benzyloxy)-2-hydroxy-1-methyl-3[(E)-isobutylidene]quinolizidine (4), which was synthesized according to the intramolecular nickel(II)/chromium(II)-mediated cyclization of the N-(iodoalkenyl)aldehyde 2. Compound 4 was converted to the acetate and subjected to reduction with lithium in ammonia, whereupon deprotection of the O-benzyl group and removal of the acetoxyl group occurred in a single operation to afford (+)-homopumiliotoxin 223G. The same sequence using (+/-)-4 was applied to the synthesis of racemic 223G;. Gas chromatography of a sample of racemic 223G showed no separation into enantiomers on four different cyclodextrin-based chiral GC columns. We found, however, that the O-acetates of (+/-)-223G gave a nearly baseline separation on either a beta-cyclodextrin column or a permethylated P-cyclodextrin column. The O-acetate of synthetic (+)-223G was identical on either of these two columns, with the first eluting O-acetate from acetylated (+/-)-223G and also with the acetylated 223G present in a frog skin extract, thus allowing us to confirm unambiguously the 1S,9aS absolute configurations of natural 223G. C1 Tokyo Univ Pharm & Life Sci, Sch Pharm, Hachioji, Tokyo 1920392, Japan. NIH, Bioorgan Chem Lab, Bethesda, MD 20892 USA. RP Kibayashi, C (reprint author), Tokyo Univ Pharm & Life Sci, Sch Pharm, Hachioji, Tokyo 1920392, Japan. NR 11 TC 16 Z9 16 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0163-3864 J9 J NAT PROD JI J. Nat. Prod. PD AUG PY 2000 VL 63 IS 8 BP 1157 EP 1159 DI 10.1021/np990641b PG 3 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 349QF UT WOS:000089056600024 PM 10978217 ER PT J AU Charan, RD Munro, MHG O'Keefe, BR Sowder, RC McKee, TC Currens, MJ Pannell, LK Boyd, MR AF Charan, RD Munro, MHG O'Keefe, BR Sowder, RC McKee, TC Currens, MJ Pannell, LK Boyd, MR TI Isolation and characterization of Myrianthus holstii lectin, a potent HIV-1 inhibitory protein from the plant Myrianthus holstii SO JOURNAL OF NATURAL PRODUCTS LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; NETTLE URTICA-DIOICA; ENVELOPE GLYCOPROTEIN GP120; AGGLUTININ; ARBOREUS; MANNOSE; INVITRO; JACALIN; FAMILY; ACID AB Aqueous extracts from the African plant Myrianthus holstii potently inhibited the infection of the T-lymphoblastoid cell line, CEM-SS; by human immunodeficiency virus-1(RF) (HIV-1(RF)). The active constituent, M. holstii lectin (MHL), was purified by LH-20 column chromatography and reversed phase HPLC. MHL, a 9284-Da cysteine-rich protein, was characterized by amino acid analysis, N-terminal sequencing, ESIMS, and matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry. Pure MHL had anti-MN activity, with an EC50 value of 150 nM. Delaying the addition of MHL for up to 8 h after initial exposure of CEM-SS cells to virus did not result in loss of the antiviral activity; however, if addition of the compound was delayed for 16 h or more, there was a marked decrease in the antiviral activity. MHL bound to a virus-free, soluble form of the viral envelope protein gp120 but did not inhibit the subsequent binding to a cell-free, soluble form of the cellular receptor CD4. C1 NCI, Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag,Dev Therapeut Program, Lab Drug Discovery Res & Dev, Frederick, MD 21702 USA. RP McKee, TC (reprint author), NCI, Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag,Dev Therapeut Program, Lab Drug Discovery Res & Dev, Bldg 1052,Room 121, Frederick, MD 21702 USA. NR 30 TC 37 Z9 41 U1 1 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0163-3864 J9 J NAT PROD JI J. Nat. Prod. PD AUG PY 2000 VL 63 IS 8 BP 1170 EP 1174 DI 10.1021/np000039h PG 5 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 349QF UT WOS:000089056600029 PM 10978222 ER PT J AU Fauerbach, JA Lawrence, JW Schmidt, CW Munster, AM Costa, PT AF Fauerbach, JA Lawrence, JW Schmidt, CW Munster, AM Costa, PT TI Personality predictors of injury-related posttraumatic stress disorder SO JOURNAL OF NERVOUS AND MENTAL DISEASE LA English DT Article ID STRUCTURED CLINICAL INTERVIEW; TRAUMATIC EVENTS; RISK-FACTORS; NEO-PI; PTSD; SAMPLE; COMORBIDITY; ADJUSTMENT; MORBIDITY; VETERANS AB This longitudinal, cohort study examined the effect of personality traits on the emergence of posttraumatic stress disorder (PTSD) in a recently traumatized, civilian, mixed-gender sample with significant injuries. Burn survivors (N = 70) were administered the NEO-Personality Inventory (NEO-PI) and the Structured Clinical Interview for DSM III-R (SCID) at hospital discharge and readministered the SCID 4 and 12 months later. Overall, the sample of burn survivors scored significantly higher on neuroticism and extraversion and lower on openness, agreeableness, and conscientiousness relative to a normative national sample. Furthermore, multivariate analysis of variance revealed that PTSD symptom severity groups (i.e., single symptom, multiple symptoms, subthreshold PTSD, PTSD) were differentially related to neuroticism and extraversion. Planned comparisons indicated that neuroticism was higher and extraversion was lower in those who developed PTSD compared with those who did not develop PTSD. C1 Johns Hopkins Univ, Sch Med,Baltimore Reg Burn Ctr, Dept Psychiat & Behav Sci, Johns Hopkins Bayview Med Ctr, Baltimore, MD 21224 USA. Johns Hopkins Univ, Sch Med,Baltimore Reg Burn Ctr, Dept Surg, Johns Hopkins Bayview Med Ctr, Baltimore, MD 21224 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Fauerbach, JA (reprint author), Johns Hopkins Univ, Sch Med,Baltimore Reg Burn Ctr, Dept Psychiat & Behav Sci, Johns Hopkins Bayview Med Ctr, 4940 Eastern Ave, Baltimore, MD 21224 USA. OI Costa, Paul/0000-0003-4375-1712 NR 36 TC 102 Z9 114 U1 1 U2 12 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3018 J9 J NERV MENT DIS JI J. Nerv. Ment. Dis. PD AUG PY 2000 VL 188 IS 8 BP 510 EP 517 DI 10.1097/00005053-200008000-00006 PG 8 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 345NF UT WOS:000088821500006 PM 10972570 ER PT J AU Tanner, SL Franzen, R Jaffe, H Quarles, RH AF Tanner, SL Franzen, R Jaffe, H Quarles, RH TI Evidence for expression of some microtubule-associated protein 1B in neurons as a plasma membrane glycoprotein SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE axolemma; dorsal root ganglion neurons; glycoprotein; membrane protein; microtubule-associated protein; neurons ID MYELIN-ASSOCIATED GLYCOPROTEIN; TANDEM MASS-SPECTROMETRY; CENTRAL-NERVOUS-SYSTEM; PHOSPHORYLATED ISOFORM; REGENERATING AXONS; IN-VITRO; MAP1B; RAT; CELLS; BRAIN AB Microtubule-associated protein (MAP) 1B is a high-molecular-weight cytoskeletal protein that is abundant in developing neuronal processes and appears to be necessary for axonal growth. Various biochemical and immunocytochemical results are reported, indicating that a significant fraction of MAP1B is expressed as an integral membrane glycoprotein in vesicles and the plasma membrane of neurons. MAP1B is present in microsomal fractions isolated from developing rat brain and fractionates across a sucrose gradient in a manner similar to synaptophysin, a well-known vesicular and plasma membrane protein. MAP1B is also in axolemma-enriched fractions (AEFs) isolated from myelinated axons of rat brain. MAP1B in AEFs and membrane fractions from cultured dorsal root ganglion neurons (DRGNs) remains membrane-associated following high-salt washes and contains sialic acid. Furthermore, MAP1B in intact DRGNs is readily degraded by extracellular trypsin and is labeled by the cell surface probe sulfosuccinimidobiotin. Immunocytochemical examination of DRGNs shows that MAP1B is concentrated in vesicle-rich varicosities along the length of axons. Myelinated peripheral nerves immunostained for MAP1B show an enrichment at the axonal plasma membrane. These observations demonstrate that some of the MAP1B in developing neurons is an integral plasma membrane glycoprotein. C1 NINDS, Myelin & Brain Dev Sect, Lab Mol & Cellular Neurobiol, NIH, Bethesda, MD 20892 USA. NINDS, Prot Peptide Sequencing Facil, NIH, Bethesda, MD 20892 USA. RP Quarles, RH (reprint author), NINDS, Myelin & Brain Dev Sect, Lab Mol & Cellular Neurobiol, NIH, Bldg 49,Room 2A28, Bethesda, MD 20892 USA. NR 51 TC 21 Z9 21 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD AUG PY 2000 VL 75 IS 2 BP 553 EP 562 DI 10.1046/j.1471-4159.2000.0750553.x PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 335EX UT WOS:000088231600013 PM 10899930 ER PT J AU Xu, H Zhang, GX Wysocka, M Wu, CY Trinchieri, G Rostami, A AF Xu, H Zhang, GX Wysocka, M Wu, CY Trinchieri, G Rostami, A TI The suppressive effect of TGF-beta on IL-12-mediated immune modulation specific to a peptide Ac1-11 of myelin basic protein (MBP): a mechanism involved in inhibition of both IL-12 receptor beta 1 and beta 2 SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE cytokines; cytokine receptors; encephalomyelitis; allergic; T lymphocytes ID GROWTH-FACTOR-BETA; INTERFERON-GAMMA PRODUCTION; CELL STIMULATORY FACTOR; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; CD4+ T-CELLS; HUMAN TUBERCULOSIS; CHAIN EXPRESSION; ORAL TOLERANCE; IN-VIVO; INTERLEUKIN-12 AB Transforming growth factor (TGF)-beta exerts a counter-regulatory effect on interleukin (IL)-12-mediated immune modulation. The underlying mechanism is not fully understood. Here we demonstrate that the expression of IL-12R beta 1 and IL-12R beta 2 in MBP peptide Acl-11-primed splenocytes is upregulated upon antigen stimulation. TGF-beta induces an unresponsiveness of these primed splenocytes to IL-12 signaling through a mechanism involved in inhibition of both IL-12R beta 1 and beta 2. The modulation of IL-12R beta 1 and beta 2 expression by Acl-11 stimulation and TGF-beta is mainly involved in CD4+ population. These data indicate that both IL-12R beta 1 and IL-12R beta 2 expression are crucial during T cell activation. TGF-beta-induced inhibition of IL-12R expression will reduce cellular immune responses during IL-12-mediated autoimmune disease. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Penn, Med Ctr, Dept Neurol, Philadelphia, PA 19104 USA. Wistar Inst, Philadelphia, PA 19104 USA. NIAID, Clin Immunol Sect, NIH, Bethesda, MD 20892 USA. RP Rostami, A (reprint author), Univ Penn, Med Ctr, Dept Neurol, 3400 Spruce St, Philadelphia, PA 19104 USA. FU NIAID NIH HHS [AI46358]; NINDS NIH HHS [NS11037] NR 44 TC 13 Z9 13 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 1 PY 2000 VL 108 IS 1-2 BP 53 EP 63 DI 10.1016/S0165-5728(00)00278-2 PG 11 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 338WW UT WOS:000088447100007 PM 10900337 ER PT J AU Koustova, E Sei, Y McCarty, T Espey, MG Ming, R Morse, HC Basile, AS AF Koustova, E Sei, Y McCarty, T Espey, MG Ming, R Morse, HC Basile, AS TI Accelerated development of neurochemical and behavioral deficits in LP-BM5 infected mice with targeted deletions of the IFN-gamma gene SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE interferon-gamma; neurodegeneration; mouse; HIV-1 soluble ICAM-1 ID ACQUIRED-IMMUNODEFICIENCY-SYNDROME; INTERFERON-GAMMA; MULTIPLE-SCLEROSIS; ADHESION MOLECULES; LEUKEMIA-VIRUS; ICAM-1; DEMENTIA; ENCEPHALOPATHY; EXPRESSION; SELECTIN AB Mice homozygous for a germline deletion of the interferon-gamma gene (IFN-gamma (-/-)) were infected with the LP-BM5 (BM5) retrovirus mixture to determine if the inability to produce IFN-gamma reduces collateral CNS damage associated with chronic neuroinflammation. Virus burdens in spleens and brains of infected mice were comparable, but spatial memory deficits were manifested earlier and to a greater extent in BMS/IFN-gamma (-/-) mice. The mice with spatial memory deficits showed considerable degradation of axone and microtubules, along with apoptosis of striatal neurons. These lesions were accompanied by extensive infiltration of perivascular spaces and ventricles by iNOS-positive leukocytes, and a 17-fold increase in CSF glutamate levels. Despite high levels of VCAM and ICAM expression on cerebral vasculature endothelia, the serum levels of soluble ICAM-1 were significantly decreased in BM5/IFN-gamma (-/-) mice, which may contribute to the enhanced leukocyte infiltration and subsequent neuronal damage. These results suggest that the presence of IFN-gamma is necessary at some points in the inflammatory process to protect against neurodegeneration. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Dept Anesthesiol, Bethesda, MD 20892 USA. RP Basile, AS (reprint author), NIDDK, Bioorgan Chem Lab, NIH, Bldg 8,Room 1A15,MSC 0826, Bethesda, MD 20892 USA. OI Morse, Herbert/0000-0002-9331-3705 NR 37 TC 10 Z9 10 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 1 PY 2000 VL 108 IS 1-2 BP 112 EP 121 DI 10.1016/S0165-5728(00)00258-7 PG 10 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 338WW UT WOS:000088447100014 PM 10900344 ER PT J AU Kenshalo, DR Iwata, K Sholas, M Thomas, DA AF Kenshalo, DR Iwata, K Sholas, M Thomas, DA TI Response properties and organization of nociceptive neurons in area 1 of monkey primary somatosensory cortex SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID MEDULLARY DORSAL HORN; NOXIOUS HEAT STIMULI; PULP-DRIVEN NEURONS; THERMAL DISCRIMINATION TASK; SI NEURONS; MECHANICAL STIMULATION; POLYMODAL NOCICEPTORS; CORTICAL-NEURONS; CONSCIOUS MONKEY; CEREBRAL-CORTEX AB The organization and response properties of nociceptive neurons in area 1 of the primary somatosensory cortex (SI) of anesthetized monkeys were examined. The receptive fields of nociceptive neurons were classified as either wide-dynamic-range (WDR) neurons that were preferentially responsive to noxious mechanical stimulation, or nociceptive specific (NS) that were responsive to only noxious stimuli. The cortical locations and the responses of the two classes of neurons were compared. An examination of the neuronal stimulus-response functions obtained during noxious thermal stimulation of the glabrous skin of the foot or the hand indicated that WDR neurons exhibited significantly greater sensitivity to noxious thermal stimuli than did NS neurons. The receptive fields of WDR neurons were significantly larger than the receptive fields of NS neurons. Nociceptive SI neurons were somatotopically organized. Nociceptive neurons with receptive fields on the foot were located more medial in area 1 of SI than those with receptive fields on the hand. In the foot representation, the recording sites of nociceptive neurons were near the boundary between areas 3b and 1, whereas in the hand area, there was a tendency for them to be located more caudal in area 1. The majority of nociceptive neurons were located in the middle layers (III and IV) of area 1. The fact that nociceptive neurons were not evenly distributed across the layers of area 1 suggested that columns of nociceptive neurons probably do not exist in the somatosensory cortex. In electrode tracks where nociceptive neurons were found, approximately half of all subsequently isolated neurons were also classified as nociceptive. Low-threshold mechanoreceptive (LTM) neurons were intermingled with nociceptive neurons. Both WDR and NS neurons were found in close proximity to one another. In instances where the receptive field shifted, subsequently isolated cells were also classified as nociceptive. These data suggest that nociceptive neurons in area 1 of SI are organized in vertically orientated aggregations or clusters in layers III and IV. C1 Ctr Sci Review, NIH, Bethesda, MD 20892 USA. NIDR, Pain & Neurosensory Mechanisms Branch, Bethesda, MD 20892 USA. Nihon Univ, Sch Dent, Dept Physiol, Tokyo 101, Japan. Harvard Univ, Sch Med, Div Med Sci, Neurosci Program, Boston, MA USA. RP Kenshalo, DR (reprint author), Ctr Sci Review, NIH, Bethesda, MD 20892 USA. NR 72 TC 114 Z9 115 U1 1 U2 8 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD AUG PY 2000 VL 84 IS 2 BP 719 EP 729 PG 11 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 343CT UT WOS:000088684700011 PM 10938299 ER PT J AU Krauzlis, RJ Basso, MA Wurtz, RH AF Krauzlis, RJ Basso, MA Wurtz, RH TI Discharge properties of neurons in the rostral superior colliculus of the monkey during smooth-pursuit eye movements SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID CAUDAL FASTIGIAL NUCLEUS; SHORT REACTION-TIMES; TARGET SELECTION; MACAQUE MONKEY; SUBCORTICAL CONNECTIONS; ELECTRICAL-STIMULATION; RESPONSE PROPERTIES; CHEMICAL LESIONS; EXPRESS SACCADES; FIXATION CELLS AB The intermediate and deep layers of the monkey superior colliculus (SC) comprise a retinotopically organized map for eye movements. The rostral end of this map, corresponding to the representation of the fovea, contains neurons that have been referred to as "fixation cells" because they discharge tonically during active fixation and pause during the generation of most saccades. These neurons also possess movement fields and are most active for targets close to the fixation point. Because the parafoveal locations encoded by these neurons are also important for guiding pursuit eye movements, we studied these neurons in two monkeys as they generated smooth pursuit. We found that fixation cells exhibit the same directional preferences during pursuit as during small saccades-they increase their discharge during movements toward the contralateral side and decrease their discharge during movements toward the ipsilateral side. This pursuit-related activity could be observed during saccade-free pursuit and was not predictive of small saccades that often accompanied pursuit. When we plotted the discharge rate from individual neurons during pursuit as a function of the position error associated with the moving target, we found tuning curves with peaks within a few degrees contralateral of the fovea. We compared these pursuit-related tuning curves from each neuron to the tuning curves for a saccade task from which we separately measured the visual, delay, and peri-saccadic activity. We found the highest and most consistent correlation with the delay activity recorded while the monkey viewed parafoveal stimuli during fixation. The directional preferences exhibited during pursuit can therefore be attributed to the tuning of these neurons for contralateral locations near the fovea. These results support the idea that fixation cells are the rostral extension of the buildup neurons found in the more caudal colliculus and that their activity conveys information about the size of the mismatch between a parafoveal stimulus and the currently foveated location. Because the generation of pursuit requires a break from fixation, the pursuit-related activity indicates that these neurons are not strictly involved with maintaining fixation. Conversely, because activity during the delay period was found for many neurons even when no eye movement was made, these neurons are also not obligatorily related to the generation of a movement. Thus the tonic activity of these rostral neurons provides a potential position-error signal rather than a motor command-a principle that may be applicable to buildup neurons elsewhere in the SC. C1 NEI, Sensorimotor Res Lab, Bethesda, MD 20892 USA. RP Krauzlis, RJ (reprint author), Salk Inst Biol Studies, 10010 N Torrey Pines Rd, La Jolla, CA 92037 USA. NR 100 TC 95 Z9 95 U1 0 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD AUG PY 2000 VL 84 IS 2 BP 876 EP 891 PG 16 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 343CT UT WOS:000088684700026 PM 10938314 ER PT J AU Basso, MA Krauzlis, RJ Wurtz, RH AF Basso, MA Krauzlis, RJ Wurtz, RH TI Activation and inactivation of rostral superior colliculus neurons during smooth-pursuit eye movements in monkeys SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID CORTICAL AREAS MT; OPTIC TRACT; REVERSIBLE INACTIVATION; ELECTRICAL-STIMULATION; FIXATION CELLS; DEFICITS; NUCLEUS; LESIONS; MST; MODULATION AB Neurons in the intermediate and deep layers of the rostral superior colliculus (SC) of monkeys are active during attentive fixation, small saccades, and smooth-pursuit eye movements. Alterations of SC activity have been shown to alter saccades and fixation, but similar manipulations have not been shown to influence smooth-pursuit eye movements. Therefore we both activated (electrical stimulation) and inactivated (reversible chemical injection) rostral SC neurons to establish a causal role for the activity of these neurons in smooth pursuit. First, we stimulated the rostral SC during pursuit initiation as well as pursuit maintenance. For pursuit initiation, stimulation of the rostral SC suppressed pursuit to ipsiversive moving targets primarily and had modest effects on contraversive pursuit. The effect of stimulation on pursuit varied with the location of the stimulation with the most rostral sites producing the most effective inhibition of ipsiversive pursuit. Stimulation was more effective on higher pursuit speeds than on lower and did not evoke smooth-pursuit eye movements during fixation. As with the effects on pursuit initiation, ipsiversive maintained pursuit was suppressed, whereas contraversive pursuit was less affected. The stimulation effect on smooth pursuit did not result from a generalized inhibition because the suppression of smooth pursuit was greater than the suppression of smooth eye movements evoked by head rotations (vestibular-ocular reflex). Nor was the stimulation effect due to the activation of superficial layer visual neurons rather than the intermediate layers of the SC because stimulation of the superficial layers produced effects opposite to those found with intermediate layer stimulation. Second, we inactivated the rostral SC with muscimol and found that contraversive pursuit initiation was reduced and ipsiversive pursuit was increased slightly, changes that were opposite to those resulting from stimulation. The results of both the stimulation and the muscimol injection experiments on pursuit are consistent with the effects of these activation and inactivation experiments on saccades, and the effects on pursuit are consistent with the hypothesis that the SC provides a position signal that is used by the smooth-pursuit eye-movement system. C1 NEI, Sensorimotor Res Lab, Bethesda, MD 20892 USA. RP Basso, MA (reprint author), NEI, Sensorimotor Res Lab, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 54 TC 66 Z9 66 U1 0 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD AUG PY 2000 VL 84 IS 2 BP 892 EP 908 PG 17 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 343CT UT WOS:000088684700027 PM 10938315 ER PT J AU Mori, Y Wakamori, M Oda, S Fletcher, CF Sekiguchi, N Mori, E Copeland, NG Jenkins, NA Matsushita, K Matsuyama, Z Imoto, K AF Mori, Y Wakamori, M Oda, S Fletcher, CF Sekiguchi, N Mori, E Copeland, NG Jenkins, NA Matsushita, K Matsuyama, Z Imoto, K TI Reduced voltage sensitivity of activation of P/Q-type Ca2+ channels is associated with the ataxic mouse mutation rolling Nagoya (tg(rol)) SO JOURNAL OF NEUROSCIENCE LA English DT Article DE P/Q-type Ca2+ channel; voltage sensor; gating charge; cerebellar Purkinje cells; ataxia; Ca2+ channel alpha(1A) subunit ID LONG-QT SYNDROME; HYPOKALEMIC PERIODIC PARALYSIS; FAMILIAL HEMIPLEGIC MIGRAINE; CEREBELLAR PURKINJE NEURONS; OMEGA-AGA-IVA; CALCIUM-CHANNEL; P-TYPE; FUNCTIONAL EXPRESSION; SODIUM-CHANNEL; MUTANT MOUSE AB Recent genetic analyses have revealed an important association of the gene encoding the P/Q-type voltage-dependent Ca2+ channel alpha(1A) subunit with hereditary neurological disorders. We have identified the ataxic mouse mutation, rolling Nagoya (tg(rol)), in the alpha(1A) gene that leads to a charge-neutralizing arginine-to-glycine substitution at position 1262 in the voltage sensor-forming segment S4 in repeat III. Ca2+ channel currents in acutely dissociated Purkinje cells, where P-type is the dominant type, showed a marked decrease in slope and a depolarizing shift by 8 mV of the conductance-voltage curve and reduction in current density in tg(rol) mouse cerebella, compared with those in wild-type. Compatible functional change was induced by the tg(rol) mutation in the recombinant alpha(1A) channel, indicating that a defect in voltage sensor of P/Q-type Ca2+ channels is the direct consequence of the tg(rol) mutation. Furthermore, somatic whole-cell recording of mutant Purkinje cells displayed only abortive Na+ burst activity and hardly exhibited Ca2+ spike activity in cerebellar slices. Thus, in tg(rol) mice, reduced voltage sensitivity, which may derive from a gating charge defect, and diminished activity of the P-type alpha(1A)Ca(2+) channel significantly impair integrative properties of Purkinje neurons, presumably resulting in locomotor deficits. C1 Natl Inst Physiol Sci, Dept Informat Physiol, Okazaki, Aichi 4448585, Japan. Grad Univ Adv Studies, Sch Life Sci, Okazaki, Aichi 4448585, Japan. Nagoya Univ, Sch Agr Sci, Lab Anim Management, Nagoya, Aichi 4648601, Japan. NCI, Frederick Canc Res & Dev Ctr, Mammalian Genet Lab, Adv Biosci Labs,Basic Res Program, Frederick, MD 21702 USA. RP Natl Inst Physiol Sci, Dept Informat Physiol, Okazaki, Aichi 4448585, Japan. EM moriy@nips.ac.jp NR 71 TC 122 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 2000 VL 20 IS 15 BP 5654 EP 5662 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 337JM UT WOS:000088354000015 PM 10908603 ER PT J AU Cohen, RI Chandross, KJ AF Cohen, RI Chandross, KJ TI Fibroblast growth factor-9 modulates the expression of myelin related proteins and multiple fibroblast growth factor receptors in developing oligodendrocytes SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE oligodendrocyte; fibroblast growth factor; immunostaining; Western blot; MAP kinase ID CENTRAL-NERVOUS-SYSTEM; MESSENGER-RNA; RAT-BRAIN; PROGENITOR CELLS; FACTOR FAMILY; FACTOR-IX; DIFFERENTIAL REGULATION; FGF RECEPTOR; NUCLEOTIDE 3'-PHOSPHODIESTERASE; GENE-EXPRESSION AB The effect of fibroblast growth factor (FGF)-9 on the expression of FGF receptors (FGFR) and the major myelin proteins was examined in cultures of developing rat brain oligodendrocytes (OLs), using immunological techniques. FGFR-1, -3, and -4 were expressed at all developmental stages but were not present in isolated myelin fractions. By contrast, FGFR-2 protein was predominantly localized to differentiating cells and myelin. FGF-9 altered FGFR and myelin protein levels during OL differentiation; there was increased expression of FGFR-1 and decreased levels of both FGFR-2 and myelin proteins. Further, FGF-9 stimulated mitogen-associated protein kinase (MAPK) phosphorylation. The effect of FGF-9 on MAPK, however, was transient and less robust in progenitor cells than in differentiated oligodendrocytes. The effects of FGF-9 and FGF-2 on FGFR and myelin protein levels were comparable; both up-regulated FGFR-1, and down-regulated FGFR-2, CNP, PLP and MBP. These findings suggest that FGF-9 may be important for glial cell development. Published 2000 Wiley-Liss, Inc.(dagger) C1 NINDS, NIH, Bethesda, MD 20892 USA. RP Cohen, RI (reprint author), NINDS, NIH, 9000 Rockville Pike,MSC 4160,Bldg 36,Room 5D05, Bethesda, MD 20892 USA. FU NINDS NIH HHS [NS23705] NR 67 TC 33 Z9 33 U1 0 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD AUG 1 PY 2000 VL 61 IS 3 BP 273 EP 287 DI 10.1002/1097-4547(20000801)61:3<273::AID-JNR5>3.0.CO;2-I PG 15 WC Neurosciences SC Neurosciences & Neurology GA 341HY UT WOS:000088586000005 PM 10900074 ER PT J AU Powell, SK Rao, J Rogue, E Nomizu, M Kuratomi, Y Yamada, Y Kleinman, HK AF Powell, SK Rao, J Rogue, E Nomizu, M Kuratomi, Y Yamada, Y Kleinman, HK TI Neural cell response to multiple novel sites on laminin-1 SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE laminin-1; nerve cells; peptides ID TERMINAL GLOBULAR DOMAIN; CEREBELLAR GRANULE NEURONS; NEURITE OUTGROWTH; SYNTHETIC PEPTIDES; A-CHAIN; BINDING SEQUENCES; HEPARIN-BINDING; ALPHA-1 CHAIN; GAMMA-1 CHAIN; IDENTIFICATION AB The basement membrane protein laminin-l is a potent stimulator of neurite outgrowth for a variety of neuronal cell types. Previous studies have identified neurite outgrowth activity in several distinct regions of the laminin-l molecule. In this study, 545 overlapping 12- to 14-mer synthetic peptides, corresponding to most of the amino acid sequence of the alpha 1, beta 1, and gamma 1 chains of laminin-l, were screened for cell attachment and neurite outgrowth activity using primary cultures of mouse cerebellar granule neurons and two neuronal cell lines. We identified 48 peptides derived from novel regions of the laminin-l molecule that were positive for neural cell adhesion activity. Only the cerebellar cells were found to have true neurite outgrowth activity with certain of the peptides, whereas some peptides induced short spike-like process with the cell lines. Although 23 of these peptides were active on all 3 cell types screened, 25 others showed cell-type specificity in their activity. These studies show that (1) there are multiple and distinct sites on laminin-l for cell adhesion and neurite-like outgrowth and (2) that there are neural cell-type-specific active domains. The multiple active sites found explains, in part, the potent activity of laminin-l on neurite outgrowth. Published 2000 Wiley-Liss, Inc.(dagger) C1 Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD USA. RP Kleinman, HK (reprint author), Bldg 30,Room 433,30 Convent Dr MSC-4370, Bethesda, MD 20892 USA. NR 38 TC 36 Z9 38 U1 0 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD AUG 1 PY 2000 VL 61 IS 3 BP 302 EP 312 DI 10.1002/1097-4547(20000801)61:3<302::AID-JNR8>3.0.CO;2-G PG 11 WC Neurosciences SC Neurosciences & Neurology GA 341HY UT WOS:000088586000008 PM 10900077 ER PT J AU Lonser, RR Oldfield, EH AF Lonser, RR Oldfield, EH TI Excitotoxic ablation - Response SO JOURNAL OF NEUROSURGERY LA English DT Letter ID ADVANCED PARKINSONS-DISEASE; POSTERIOR GPI PALLIDOTOMY; POSTEROVENTRAL PALLIDOTOMY; DELIVERY; MACROMOLECULES; INFUSION; BRAIN C1 Univ Utah, Hlth Sci Ctr, Salt Lake City, UT 84112 USA. NIH, Bethesda, MD 20892 USA. RP Lonser, RR (reprint author), Univ Utah, Hlth Sci Ctr, Salt Lake City, UT 84112 USA. NR 15 TC 0 Z9 0 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 2000 VL 93 IS 2 BP 365 EP 366 PG 2 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 338HY UT WOS:000088414500031 ER PT J AU Chang, HC Churchwell, MI Delclos, KB Newbold, RR Doerge, DR AF Chang, HC Churchwell, MI Delclos, KB Newbold, RR Doerge, DR TI Mass spectrometric determination of genistein tissue distribution in diet-exposed Sprague-Dawley rats SO JOURNAL OF NUTRITION LA English DT Article DE genistein; isoflavones; mass spectrometry; rats; pharmacokinetics ID ESTROGEN-RECEPTOR-BETA; SOY ISOFLAVONES; QUANTITATIVE-ANALYSIS; FEMALE RATS; IN-VITRO; PHYTOESTROGENS; EXCRETION; DAIDZEIN; METABOLISM; PLASMA AB Genistein, the principal soy isoflavone, was administered in the diet to male and female Sprague-Dawley rats as part of a multigeneration study of potential endocrine modulation. The rats were exposed to genistein in utero, through maternal milk, and as adults through postnatal d 140 via essentially isoflavone-free feed (similar to 0.5 mu g/g) fortified at 5, 100 and 500 mu g/g with genistein aglycone. Analytical methods based on liquid chromatography, mass spectrometry and the use of deuterated genistein were developed and validated for use in measuring genistein in serum and tissues. Pharmacokinetic analysis of serum genistein showed a significant difference (P < 0.001) in the elimination half-life and area under the concentration-time curve between male [2.97 +/- 0.14 h and 22.3 +/- 1.2 mu mol/(L . h), respectively] and female rats [4.26 +/- 0.29 h and 45.6 +/- 3.1 mu mol/(L . h), respectively, +/- SEM]. Endocrine-responsive tissues including brain, liver, mammary, ovary, prostate, testis, thyroid and uterus showed significant dose-dependent increases in total genistein concentration. Female liver contained the highest amount of genistein (7.3 pmol/mg tissue) and male whole brain contained the least (0.04 pmol/mg). The physiologically active aglycone form was present in tissues at fractions up to 100%, and the concentration was always greater than that observed in serum in which conjugated forms predominated (95-99%). These results for measured amounts of genistein, present as aglycone and conjugates, in putative target tissues provide a link with other studies in which blood concentrations and physiologic effects of genistein are measured. C1 Natl Ctr Toxicol Res, Div Biochem Toxicol, Jefferson, AR 72079 USA. NIEHS, Dev Endocrinol Sect, Reprod Toxicol Grp, Lab Toxicol,Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. RP Doerge, DR (reprint author), Natl Ctr Toxicol Res, Div Biochem Toxicol, Jefferson, AR 72079 USA. NR 30 TC 195 Z9 203 U1 0 U2 12 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 2000 VL 130 IS 8 BP 1963 EP 1970 PG 8 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 339TA UT WOS:000088492900015 PM 10917909 ER PT J AU Murata, M Kador, PF Sato, S AF Murata, M Kador, PF Sato, S TI Vascular endothelial growth factor (VEGF) enhances the expression of receptors and activates mitogen-activated protein (MAP) kinase of dog retinal capillary endothelial cells SO JOURNAL OF OCULAR PHARMACOLOGY AND THERAPEUTICS LA English DT Article ID GALACTOSE-FED DOGS; ALDOSE REDUCTASE INHIBITORS; TYROSINE KINASE; SIGNAL-TRANSDUCTION; PERMEABILITY FACTOR; DIABETIC-RETINOPATHY; FLT-1; KDR; IDENTIFICATION; ANGIOGENESIS AB Since the galactose-fed dog is an animal model that develops the advanced stage of proliferative retinopathy, the effects of vascular endothelial growth factor (VEGF) on cell growth, receptor expression and the activation of mitogen-activated protein (MAP) kinase pathway of dog retinal capillary endothelial cells were investigated. Dog retinal endothelial cells were cultured at 37 degrees C under 5% carbon dioxide atmosphere in CS-C medium supplemented with endothelial cell growth factor(ECGF). VEGF receptor expression was examined by RT-PCR, and activation of MAP kinase was examined with antibody against phospho-Elk-l (Ser383). When growth factors were removed from the culture medium, cell survival of dog endothelial cells was significantly reduced. Addition of VEGF protected these cells from cell death induced by growth factor starvation. VEGF also enhanced tube formation in dog endothelial cells and increased the expression of two VEGF receptors, Flt-1 and KDR/Flk-1. Cells treated with VEGF also displayed the phosphorylation of the transcription factor, Elk-1. Addition of the tyrosine kinase inhibitor, genistein, eliminated VEGF-induced cell growth and Elk-1 phosphorylation. These data confirm that cell growth and tube formation of dog retinal capillary endothelial cells are stimulated by VEGF. VEGF also increases the expression of the receptors, KDR and Flt-1, and activates the p44/42 MAP kinase pathway. C1 NEI, LOT, NIH, Bethesda, MD 20892 USA. RP Sato, S (reprint author), NEI, LOT, NIH, 10-10B09,10 Ctr Dr,MSC 1850, Bethesda, MD 20892 USA. NR 39 TC 19 Z9 20 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1080-7683 J9 J OCUL PHARMACOL TH JI J. Ocular Pharmacol. Ther. PD AUG PY 2000 VL 16 IS 4 BP 383 EP 391 DI 10.1089/jop.2000.16.383 PG 9 WC Ophthalmology; Pharmacology & Pharmacy SC Ophthalmology; Pharmacology & Pharmacy GA 345ME UT WOS:000088819100010 PM 10977134 ER PT J AU Overpeck, MD Hediger, ML Ruan, WJ Davis, WW Maurer, KR Troendle, JF Kuczmarski, RJ AF Overpeck, MD Hediger, ML Ruan, WJ Davis, WW Maurer, KR Troendle, JF Kuczmarski, RJ TI Stature, weight, and body mass among young US children born at term with appropriate birth weights SO JOURNAL OF PEDIATRICS LA English DT Article ID NUTRITION-EXAMINATION-SURVEY; DISEASE RISK-FACTORS; 3RD NATIONAL-HEALTH; GESTATIONAL-AGE; UNITED-STATES; OBESITY; GROWTH; CHILDHOOD; ADULTS; HEART AB Objective: To describe weight, stature, and body mass index (BMI) changes occurring before the age of 7 years, which may influence the prevalence of overweight in adolescence and adulthood. Methods: Regression models predicting height and weight at ages 2 months to 6.75 years were based on the third National Health and Nutrition Examination Survey. Birth certificate data were used to adjust ethnic-specific models for birth weight for gestational age. Results: Attained height is higher for non-Hispanic black children than for either non-Hispanic white or Mexican American children (P less than or equal to 5.001). Weights at 2 months, 2 years, and 6.75 years are similar among racial/ethnic groups after adjustments. Mexican American children at ages 2 to 6.75 years had higher prevalences of BMI >85th percentile than either non-Hispanic white or black children (boys = 25.6%, SE = 2.7 compared with 14.1%, SE = 1.7 and 16.5%, SE = 1.7, respectively girls = 21.9%, SE = 3.6 compared with 13.0%, SE = 1.7 and 13.7%, SE = 2.2, respectively). For non-Hispanic whites and Mexican Americans and for non-Hispanic black boys, BMI decreased slightly between ages 2 and 6.75 years; BMI for nonHispanic black girls did not. Conclusion: Size differences before the age of 7 years may influence later ethnic-specific overweight prevalence, independent of prenatal influences. C1 NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. NCI, Bethesda, MD 20892 USA. Natl Ctr Hlth Stat, Div Hlth Examinat Stat, Hyattsville, MD 20782 USA. RP Overpeck, MD (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. NR 40 TC 13 Z9 14 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 2000 VL 137 IS 2 BP 205 EP 213 DI 10.1067/mpd.2000.107163 PG 9 WC Pediatrics SC Pediatrics GA 345TZ UT WOS:000088832400014 PM 10931413 ER PT J AU Catt, KJ Olivares Reyes, AJ Zhang, M Smith, RD Hunyady, L AF Catt, KJ Olivares Reyes, AJ Zhang, M Smith, RD Hunyady, L TI Molecular mechanisms of regulation of angiotensin II receptors SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Meeting Abstract C1 NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Semmelweis Univ, Sch Med, Dept Physiol, H-1088 Budapest, Hungary. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0022-3751 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD AUG PY 2000 VL 526 SU S BP 2S EP 3S PG 2 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 355GP UT WOS:000089378200274 ER PT J AU Mari, Z Mima, T Gerloff, C Hallett, M Bodis-Wollner, I AF Mari, Z Mima, T Gerloff, C Hallett, M Bodis-Wollner, I TI Perisaccadic high frequency EEG changes in frontal and occipital regions are similar in light and dark SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Meeting Abstract ID ACTIVATION; VOLUNTARY; SACCADES; CORTEX C1 SUNY Hlth Sci Ctr, Dept Neurol, Brooklyn, NY 11203 USA. NINDS, NIH, Bethesda, MD 20892 USA. NR 8 TC 0 Z9 0 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0022-3751 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD AUG PY 2000 VL 526 SU S BP 25S EP 25S PG 1 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 355GP UT WOS:000089378200298 ER PT J AU Pando, JA Duray, P Yarboro, C Gourley, MF Klippel, JH Schumacher, HR AF Pando, JA Duray, P Yarboro, C Gourley, MF Klippel, JH Schumacher, HR TI Synovitis occurs in some clinically normal and asymptomatic joints in patients with early arthritis SO JOURNAL OF RHEUMATOLOGY LA English DT Article DE rheumatoid arthritis; reactive arthritis; synovitis; knee ID RHEUMATOID-ARTHRITIS; CHLAMYDIA-TRACHOMATIS; TISSUE; DNA AB Objective. To determine if clinically asymptomatic knee joints in patients with recent onset arthritis reveal histological evidence of synovitis. Methods, As part of a prospective study of patients with synovitis of less than one year duration, we performed blind needle biopsies on the knees of 20 patients who had synovitis elsewhere but no symptoms or detectable swelling or tenderness of the biopsied joint. Results, Histologic evidence of synovitis was observed in 11 knees (55%). All patients with synovitis had evidence of synovial lining cell hyperplasia, increased vascularity, and lymphocytic infiltrates. Five of 6 patients with rheumatoid arthritis (RA) and 5 of 8 with undifferentiated arthritis had histological evidence of synovitis. but none of the 5 with reactive arthritis (ReA) had synovitis in the asymptomatic joints. Histologic evidence of synovitis persisted in some after clinical resolution of previous pain and swelling, while it occurred in others with no history of previous involvement of that knee. Conclusion. Even asymptomatic joints in patients with RA and undifferentiated arthritis of recent onset reveal histologic signs of synovitis. The earliest changes may occur before symptoms. Histologic changes also persist after resolution of previous early symptoms. Evidence of inflammation was not present in asymptomatic joints in our 5 patients diagnosed with ReA. C1 NIAMS, Arthrit & Rheumatism Branch, NIH, Bethesda, MD USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Univ Penn, Div Rheumatol, Philadelphia, PA 19104 USA. Vet Affairs Med Ctr, Philadelphia, PA USA. RP Pando, JA (reprint author), 1305 Savannah Rd, Lewes, DE 19951 USA. NR 23 TC 29 Z9 30 U1 0 U2 1 PU J RHEUMATOL PUBL CO PI TORONTO PA 920 YONGE ST, SUITE 115, TORONTO, ONTARIO M4W 3C7, CANADA SN 0315-162X J9 J RHEUMATOL JI J. Rheumatol. PD AUG PY 2000 VL 27 IS 8 BP 1848 EP 1854 PG 7 WC Rheumatology SC Rheumatology GA 340PM UT WOS:000088543500007 PM 10955323 ER PT J AU Gannot, G Lancaster, HE Fox, PC AF Gannot, G Lancaster, HE Fox, PC TI Clinical course of primary Sjogren's syndrome: Salivary, oral, and serologic aspects SO JOURNAL OF RHEUMATOLOGY LA English DT Article DE Sjogren's syndrome; saliva; lymphoma; autoimmune disease ID LYMPHOMA AB Objective. To assess changes in symptoms and signs, salivary function, serologic activity, and disease progression in primary Sjogren's syndrome (SS). Methods. Treatment records on 80 patients seen in clinic and diagnosed with primary SS by defined criteria were reviewed. Forty-nine patients were evaluated at least twice a minimum of 5 years (mean 7 years) apart. Salivary flow rates from each of the major salivary glands and laboratory values were obtained. A structured interview with questions pertaining to signs and symptoms of primary SS was given and a physical examination was performed. An additional 26 patients completed a followup questionnaire by mail and their current medical records were obtained for review. For this group, the followup period was a mean of 10 years after their initial evaluation. Five patients were deceased. Results. The patients seen twice showed relative stability in their salivary measurements and in their serologic values. The subjective sicca symptoms of oral and ocular dryness among the 75 surviving patients remained prominent. Very few individuals developed another connective tissue disease. therefore evolving into secondary SS, Among the 80 patients, 6 cases of B cell lymphoma were recognized during the followup period. Conclusion. Although it is nor a benign condition, primary SS is a very slowly progressing disease without rapid deterioration in salivary function, systemic markers of disease activity, or dramatic changes in symptoms, with the exception of a high incidence of lymphoma. C1 Natl Inst Dent & Craniofacial Res, Clin Invest Sect, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD USA. RP Fox, PC (reprint author), Amarillo Biosci Inc, 6509 Seven Locks Rd, Cabin John, MD 20818 USA. NR 14 TC 38 Z9 42 U1 0 U2 0 PU J RHEUMATOL PUBL CO PI TORONTO PA 920 YONGE ST, SUITE 115, TORONTO, ONTARIO M4W 3C7, CANADA SN 0315-162X J9 J RHEUMATOL JI J. Rheumatol. PD AUG PY 2000 VL 27 IS 8 BP 1905 EP 1909 PG 5 WC Rheumatology SC Rheumatology GA 340PM UT WOS:000088543500015 PM 10955331 ER PT J AU Vitiello, B AF Vitiello, B TI Stimulant treatment for children: A community perspective - Commentary SO JOURNAL OF THE AMERICAN ACADEMY OF CHILD AND ADOLESCENT PSYCHIATRY LA English DT Editorial Material ID DEFICIT-HYPERACTIVITY DISORDER; ATTENTION-DEFICIT/HYPERACTIVITY DISORDER; PRIMARY-CARE; METHYLPHENIDATE; TRENDS; ADHD C1 NIMH, Child & Adolescent Treatment & Prevent Intervent, Bethesda, MD 20892 USA. RP Vitiello, B (reprint author), NIMH, Child & Adolescent Treatment & Prevent Intervent, Room 7147,6001 Execut Blvd,MSC 9633, Bethesda, MD 20892 USA. NR 13 TC 2 Z9 2 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0890-8567 J9 J AM ACAD CHILD PSY JI J. Am. Acad. Child Adolesc. Psychiatr. PD AUG PY 2000 VL 39 IS 8 BP 992 EP 994 DI 10.1097/00004583-200008000-00013 PG 3 WC Psychology, Developmental; Pediatrics; Psychiatry SC Psychology; Pediatrics; Psychiatry GA 338JF UT WOS:000088415300013 ER PT J AU Stiffman, AR Horwitz, SM Hoagwood, K Compton, W Cottler, L Bean, DL Narrow, WE Weisz, JR AF Stiffman, AR Horwitz, SM Hoagwood, K Compton, W Cottler, L Bean, DL Narrow, WE Weisz, JR TI The Service Assessment for Children and Adolescents (SACA): Adult and child reports SO JOURNAL OF THE AMERICAN ACADEMY OF CHILD AND ADOLESCENT PSYCHIATRY LA English DT Article DE mental health services; service use; adolescent mental health ID PSYCHOPATHOLOGY RATING-SCALES; INTERRATER AGREEMENT; BEHAVIORAL-PROBLEMS; REFERRED CHILDREN; PARENT RATINGS; INTERVIEW; DEPRESSION; SYMPTOMS; INFORMANTS; SCHEDULE AB Objective: To describe differences in parent-child responses to the Service Assessment for Children and Adolescents (SACA). Method: Studies were done at UCLA and Washington University based on service-using and community subjects drawn from community households or public school student lists, respectively. Results are presented for 145 adult-youth pairs in which the youth was II or older. Results: The SAGA adult-youth correspondence for lifetime use of any services, inpatient services, outpatient services, and school services ranged from fair to excellent (kappa = 0.43-0.86, with most at 0.61 or greater). Similarly, the SAGA showed a good to excellent correspondence for services that had been used in the preceding year (kappa = 0.45-0.77, with most greater than 0.50). The parent-youth correspondence for use of specific service settings in the above generic categories ranged from poor to excellent (kappa = 0.25-0.83, with half at 0.50 or greater). Conclusions: The SAGA has better adult-youth correspondence than any service use questionnaire with published data, indicating that both adult and youth reports are not needed for all research on mental health services. This is especially encouraging news for researchers working with high-risk youth populations, in which a parent figure is often not available. C1 Washington Univ, George Warren Brown Sch Social Work, St Louis, MO 63130 USA. Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, New Haven, CT 06510 USA. NIMH, Child Serv, Bethesda, MD 20892 USA. NIMH, Adolescent Mental Hlth Serv, Bethesda, MD 20892 USA. Washington Univ, Sch Med, Dept Psychiat, St Louis, MO 63110 USA. Univ Calif Los Angeles, Los Angeles, CA 90024 USA. RP Stiffman, AR (reprint author), Washington Univ, George Warren Brown Sch Social Work, St Louis, MO 63130 USA. FU NIMH NIH HHS [U01 MH/HD54280, U01 MH/HD54282, U01 MH/HD54293] NR 30 TC 85 Z9 85 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0890-8567 J9 J AM ACAD CHILD PSY JI J. Am. Acad. Child Adolesc. Psychiatr. PD AUG PY 2000 VL 39 IS 8 BP 1032 EP 1039 DI 10.1097/00004583-200008000-00019 PG 8 WC Psychology, Developmental; Pediatrics; Psychiatry SC Psychology; Pediatrics; Psychiatry GA 338JF UT WOS:000088415300019 PM 10939232 ER PT J AU Lebowitz, NE Bella, JN Roman, MJ Liu, JE Fishman, DP Paranicas, M Lee, ET Fabsitz, RR Welty, TK Howard, BV Devereux, RB AF Lebowitz, NE Bella, JN Roman, MJ Liu, JE Fishman, DP Paranicas, M Lee, ET Fabsitz, RR Welty, TK Howard, BV Devereux, RB TI Prevalence and correlates of aortic regurgitation in American Indians: The strong heart study SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID APPETITE-SUPPRESSANT DRUGS; STRUCTURALLY NORMAL HEARTS; VALVULAR REGURGITATION; DOPPLER ECHOCARDIOGRAPHY; CARDIOVASCULAR-DISEASE; OBESE PATIENTS; RISK-FACTORS; ABNORMALITIES; HYPERTENSION; ASSOCIATION AB OBJECTIVES We sought to determine the prevalence and correlates of aortic regurgitation (AR) in a population-based sample group. BACKGROUND Concern over induction of AR by weight loss medication highlights the importance of assessing the prevalence and correlates of AR in unselected patient groups. METHODS Aortic regurgitation was assessed by color flow Doppler echocardiography in 3,501 American Indian participants age 47 to 81 years during the second Strong Heart Study. RESULTS Mild (If) AR was present in 7.3%, 2+ AR in 2.4% and 3+ to 4+ AR in 0.3% of participants, more frequently in those greater than or equal to 60 years old than in those <60 years old (14.4% vs. 5.8%, p < 0.001); AR was unrelated to gender. Compared with participants without AR, those with mild AR had a lower body mass index (p < 0.004) and higher systolic pressure (p < 0.003). Participants with AR had larger aortic root diameters (3.6 +/- 0.4 vs. 3.4 +/- 0.4 cm, p < 0.001), higher creatinine levels (1.3 +/- 1.3 vs. 1.0 +/- 1.0 mg/dl, p < 0.001) and higher urine albumin/creatinine levels (3.6 +/- 2.3 vs. 3.3 +/- 2.0 log, p < 0.001), as well as higher prevalences of aortic stenosis (AS) or mitral stenosis (MS) (p < 0.001). Regression analysis showed that AR was independently related to older age and larger aortic roots (p < 0.0001), AS and absence of diabetes (p = 0.002), MS (p = 0.003) and higher log urine albumin/creatinine (p = 0.005). CONCLUSIONS Aortic regurgitation occurred in 10% of a sample group of middle-aged to older adults and was related to older age, larger aortic root diameter, aortic and mitral stenosis and albuminuria. There was no association of AR with being overweight and a negative association of AR with diabetes. U Am Cell Cardiol 2000;36:461-7) (C) 2000 by the American College of Cardiology. C1 Cornell Univ, Med Ctr, New York Hosp, Dept Med, New York, NY 10021 USA. Univ Oklahoma, Hlth Sci Ctr, Sch Publ Hlth, Oklahoma City, OK 73190 USA. NHLBI, Bethesda, MD 20892 USA. Aberdeen Area Tribal Chairmans Hlth Board, Rapid City, SD USA. Medlant Res Inst, Washington, DC USA. RP Devereux, RB (reprint author), New York Presbyterian Hosp, Weill Cornell Med Ctr, Div Cardiol, Box 222,525 E 68th St, New York, NY 10021 USA. FU NHLBI NIH HHS [U01-HL41654, U01-HL41652, U01-HL41642] NR 36 TC 63 Z9 64 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD AUG PY 2000 VL 36 IS 2 BP 461 EP + DI 10.1016/S0735-1097(00)00744-0 PG 8 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 339RN UT WOS:000088491800016 PM 10933358 ER PT J AU Padayatty, SJ Levine, M AF Padayatty, SJ Levine, M TI Reevaluation of ascorbate in cancer treatment: Emerging evidence, open minds and serendipity SO JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION LA English DT Editorial Material DE cancer; ascorbate; plasma concentration; intravenous; cytotoxicity; vitamin C ID TERMINAL HUMAN CANCER; DOSE VITAMIN-C; SUPPLEMENTAL ASCORBATE; SUPPORTIVE TREATMENT; SURVIVAL TIMES; ACID THERAPY; CELL-LINES; PROLONGATION; DERIVATIVES; VOLUNTEERS AB Some clinicians and alternative therapy practitioners advocate megadose intravenous and oral ascorbate treatment of cancer. Randomized control studies using oral ascorbate showed no benefit. Recent data show that intravenous but nor oral administration of ascorbate can produce millimolar plasma concentrations, which are toxic to many cancer cell lines. We propose that ascorbate treatment of cancer should be reexamined by rigorous scientific scrutiny in the light of new evidence. C1 NIDDKD, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RP Padayatty, SJ (reprint author), NIDDKD, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RI Padayatty, Sebastian/A-8581-2012 OI Padayatty, Sebastian/0000-0001-8758-3170 NR 26 TC 50 Z9 54 U1 2 U2 4 PU AMER COLL NUTRITION PI NEW YORK PA C/O HOSP. JOINT DIS. 301 E. 17TH ST., NEW YORK, NY 10003 USA SN 0731-5724 J9 J AM COLL NUTR JI J. Am. Coll. Nutr. PD AUG PY 2000 VL 19 IS 4 BP 423 EP 425 PG 3 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 344DF UT WOS:000088741800001 PM 10963459 ER PT J AU Foley, DJ Masaki, KH Ross, GW White, LR AF Foley, DJ Masaki, KH Ross, GW White, LR TI Driving cessation in older men with incident dementia SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Article DE driving; Alzheimer's disease; dementia; mobility; older men; epidemiology ID ALZHEIMERS-DISEASE; RISK; AGE AB OBJECTIVE: To determine the prevalence and cessation of driving among order men with incident dementia in the Honolulu-Asia Aging Study. DESIGN: Retrospective cohort data from a community-based study of incident dementia. SETTING: The Honolulu Heart Program and the Honolulu-Asia Aging Study. PARTICIPANTS: A total of 643 men who were evaluated for the incidence of Alzheimer's disease or other dementia between the fourth and the fifth examination of the Honolulu Heart Program. MEASUREMENTS: Driving history, diagnosis of dementia, grip strength, walking speed, standing balance test, interviewer's rating of vision status, and the neurologist's notes on mentions of driving behavior from informal interviews with a caregiver or family informant. RESULTS: The prevalence of driving declined dramatically with level of cognitive functioning. Among 162 men evaluated and found to have normal cognitive functioning, 78% still drove, compared with 62% of 287 men with poor cognitive functioning but no clinical dementia, 46% of 96 men with a new diagnosis of very mild dementia (Clinical Dementia Rating = 0.5), and 22% of 98 men with a new diagnosis of mild dementia (CDR = 1). Only one of 23 men diagnosed with moderate or more severe staged incident dementia (CDR > 1) was driving. About 10% of the 59 demented persons still driving relied on co-pilots, and only one driver was reported as involved in a crash according to a review of the neurologists' notes. CONCLUSIONS: Incident dementia is a major cause of driving cessation. Based on these data, we estimate that approximately 4% of male drivers aged 75 years and older nationwide (about 175,000 men) have dementia. This number will increase with the projected growth of drivers aged 75 years and older. C1 NIA, Epidemiol Demog & Biometry Program, NIH, Bethesda, MD 20892 USA. Kuakini Med Ctr, Hololulu Asia Aging Study, Honolulu Heart Program, Honolulu, HI 96817 USA. Univ Hawaii, John A Burns Sch Med, Honolulu, HI 96822 USA. Dept Vet Affairs, Honolulu, HI USA. RP Foley, DJ (reprint author), NIA, Epidemiol Demog & Biometry Program, NIH, 7201 Wisconsin Ave,Gateway Bldg,Suite 3C-309, Bethesda, MD 20892 USA. FU NHLBI NIH HHS [N01-HC-05102]; NIA NIH HHS [N01-AG-4-2149] NR 20 TC 39 Z9 40 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 BP 928 EP 930 PG 3 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000008 PM 10968296 ER PT J AU Arseven, A McDermott, M O'Brien, E Liu, K Guralnik, J AF Arseven, A McDermott, M O'Brien, E Liu, K Guralnik, J TI Fall risk is not increased among older men and women with lower extremity arterial disease SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. Northwestern Univ, Sch Med, Chicago, IL USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P156 BP S50 EP S50 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000222 ER PT J AU Cheh, A Reuben, D Harris, T Seeman, TE AF Cheh, A Reuben, D Harris, T Seeman, TE TI Peripheral blood markers of inflammation predict mortality and functional decline among high functioning community-dwelling older persons SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Univ Calif Los Angeles, Los Angeles, CA USA. NIA, Bethesda, MD 20892 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P459 BP S123 EP S123 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000514 ER PT J AU Edmond, J Metz, D Harman, S Barofsky, I Pabst, K Munzer, T Christmas, C Sorkin, J Blackman, M AF Edmond, J Metz, D Harman, S Barofsky, I Pabst, K Munzer, T Christmas, C Sorkin, J Blackman, M TI Effects of growth hormone and/or sex steroid administration on health-related quality of life in healthy aged individuals SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Johns Hopkins Univ, Baltimore, MD USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P283 BP S81 EP S81 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000348 ER PT J AU Hu, PF Seeman, TE Harris, TB Reuben, DB AF Hu, PF Seeman, TE Harris, TB Reuben, DB TI Do inflammation or malnutrition explain the low cholesterol mortality association in community-dwelling older persons? MacArthur Studies of Successful Aging SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P456 BP S122 EP S122 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000512 ER PT J AU Kiel, DP Harris, TB Visser, M Hannan, MT Wilson, PWF AF Kiel, DP Harris, TB Visser, M Hannan, MT Wilson, PWF TI Leptin and bone mineral density (BMD) in older men and women SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Hebrew Rehabil Ctr Aged, Res & Training Inst, Boston, MA 02131 USA. NIA, Bethesda, MD 20892 USA. Inst Res Extramural Med, Amsterdam, Netherlands. Harvard Univ, Sch Med, Div Aging, Boston, MA 02115 USA. Boston Univ, Framingham, MA USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P295 BP S84 EP S84 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000360 ER PT J AU Kramarow, E Lentzner, H Rooks, R Weeks, J Saydah, S AF Kramarow, E Lentzner, H Rooks, R Weeks, J Saydah, S TI The health of older Americans: Highlights from health and aging chartbook SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Natl Ctr Hlth Stat, Hyattsville, MD 20782 USA. NIA, Bethesda, MD 20892 USA. Johns Hopkins Univ, Baltimore, MD 21218 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P345 BP S96 EP S96 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000408 ER PT J AU Ling, S Bandeen-Roche, K Fan, MY Bathon, J Guralnik, J Fried, L AF Ling, S Bandeen-Roche, K Fan, MY Bathon, J Guralnik, J Fried, L TI Disability associated with comorbid osteoarthritis (OA) and heart disease (HD) SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Johns Hopkins Univ, Baltimore, MD USA. NIA, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P97 BP S35 EP S35 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000164 ER PT J AU Masaki, K Yano, K Schatz, I Chen, R Foley, D Rodriguez, B Blanchette, P Curb, JD AF Masaki, K Yano, K Schatz, I Chen, R Foley, D Rodriguez, B Blanchette, P Curb, JD TI Orthostatic hypotension predicts five-year incidence of coronary heart disease in elderly men: The honolulu heart program. SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. Univ Hawaii, Manoa, HI 96822 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P418 BP S113 EP S113 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000476 ER PT J AU Munzer, T Harman, SM Metz, D Pabst, K Barofsky, I Christmas, C Roy, T Tobin, J Sorkin, J Blackman AF Munzer, T Harman, SM Metz, D Pabst, K Barofsky, I Christmas, C Roy, T Tobin, J Sorkin, J Blackman TI Effects of growth hormone (GH) and/or sex steroids (SS) on the relationship between body fat and perceived body image in healthy aged individuals SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P154 BP S49 EP S49 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000220 ER PT J AU Newman, AB Yanez, D Duxbury, AS Enright, PL Fried, LP Harris, TA AF Newman, AB Yanez, D Duxbury, AS Enright, PL Fried, LP Harris, TA TI Weight loss but not gain predicts mortality in older adults SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Univ Pittsburgh, Pittsburgh, PA USA. Univ Washington, Seattle, WA 98195 USA. Univ Alabama, Birmingham, AL USA. Univ Arizona, Tucson, AZ USA. Johns Hopkins Univ, Baltimore, MD USA. NIA, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P270 BP S78 EP S78 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000335 ER PT J AU Reuben, D Ferrucci, L Wallace, R Tracy, R Corti, MC Heimovitz, H Harris, TB AF Reuben, D Ferrucci, L Wallace, R Tracy, R Corti, MC Heimovitz, H Harris, TB TI The prognostic value of serum albumin in healthy older persons with low and high serum IL-6 levels SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Univ Calif Los Angeles, Los Angeles, CA USA. Italian Nat Res Council Aging, Florence, Italy. Univ Iowa, Iowa City, IA USA. Acad Clin Lab Phys & Scientists, Burlington, VT USA. Univ Padua, Padua, Italy. Styel, Bethesda, MD USA. NIA, Bethesda, MD 20892 USA. NR 0 TC 3 Z9 3 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA A36 BP S10 EP S10 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000066 ER PT J AU Volpato, S Leveille, S Corti, M Harris, T Guralnik, J AF Volpato, S Leveille, S Corti, M Harris, T Guralnik, J TI The value of serum albumin and HDL-cholesterol in defining mortality risk in older person with low serum cholesterol SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. Unita Valutaz Geriatr, Camposampiero, Italy. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P293 BP S84 EP S84 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000358 ER PT J AU Zieman, S Lakatta, E Fleg, J Gerstenblith, G O'Connor, F Brant, L Bos, A AF Zieman, S Lakatta, E Fleg, J Gerstenblith, G O'Connor, F Brant, L Bos, A TI Pulse pressure rises exponentially with age by longitudinal analysis SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Meeting Abstract C1 Johns Hopkins Univ, Baltimore, MD USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. 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 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD AUG PY 2000 VL 48 IS 8 MA P424 BP S115 EP S115 PG 1 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 346TK UT WOS:000088887000482 ER PT J AU Doi, SQ Jacot, TA Sellitti, DF Hirszel, P Hirata, MH Striker, GE Striker, LJ AF Doi, SQ Jacot, TA Sellitti, DF Hirszel, P Hirata, MH Striker, GE Striker, LJ TI Growth hormone increases inducible nitric oxide synthase expression in mesangial cells SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article ID ORNITHINE DECARBOXYLASE ACTIVITY; TRANSGENIC MICE; PROGRESSIVE GLOMERULOSCLEROSIS; EXPERIMENTAL GLOMERULONEPHRITIS; NEPHROTOXIC NEPHRITIS; KIDNEY HYPERTROPHY; GH ANTAGONIST; MESSENGER-RNA; RAT; INHIBITION AB Mice transgenic for bovine growth hormone (GH) develop progressive glomerulosclerosis. However, the proximal signaling events that lead to increased matrix deposition in this pathologic condition are still unclear. Components of the L-arginine metabolic pathway, especially inducible nitric oxide (NO) synthase (iNOS), ornithine aminotransferase (OAT), and ornithine decarboxylase (ODC), have been associated with glomerular scarring. In this study, mesangial cells were treated with GH, and the expression of NOS, ODC, and OAT was determined using reverse transcription-PCR, In addition, nitrite accumulation in the conditioned media of mesangial cell cultures was measured in the presence or absence of GH. The findings revealed that GH increased iNOS transcript levels in a dose-dependent manner, with the highest levels being attained at GH concentrations of 20 to 50 ng/ml. The GH-induced increase in iNOS transcript levels was accompanied by a significant increase in nitrite concentrations in conditioned media, which was blocked by the addition of L-N-G-monomethylarginine. The effect of GH (50 ng/ml) in eliciting nitrite production was as potent as that of bacterial lipopolysaccharide (10 mu g/ml). The expression of OAT and ODC, in contrast, was not altered at any of the GH concentrations tested. GH receptor mRNA was also expressed by mesangial cells, independently of the GH concentration present in the cell culture medium. These data indicate that GH may interact with its receptor to regulate the L-arginine/NO pathway in mesangial cells, by directly modulating iNOS expression and NO production, without altering the arginase/OAT/ODC pathway. C1 Uniformed Serv Univ Hlth Sci, Dept Med, Div Nephrol, Bethesda, MD 20814 USA. NIDDKD, Renal Cell Biol Sect, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. Univ Sao Paulo, Sch Pharmaceut Sci, Sao Paulo, Brazil. Univ Miami, Sch Med, Dept Med, Miami, FL USA. RP Doi, SQ (reprint author), Uniformed Serv Univ Hlth Sci, Dept Med, Div Nephrol, 4301 Jones Bridge Rd,A-3068, Bethesda, MD 20814 USA. RI Hirata, Mario/C-9718-2013 NR 44 TC 26 Z9 26 U1 0 U2 1 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 2000 VL 11 IS 8 BP 1419 EP 1425 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA 337CY UT WOS:000088340900006 PM 10906155 ER PT J AU Kimmel, PL Peterson, RA Weihs, KL Shidler, N Simmens, SJ Alleyne, S Cruz, I Yanovski, JA Veis, JH Phillips, TM AF Kimmel, PL Peterson, RA Weihs, KL Shidler, N Simmens, SJ Alleyne, S Cruz, I Yanovski, JA Veis, JH Phillips, TM TI Dyadic relationship conflict, gender, and mortality in urban hemodialysis patients SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article; Proceedings Paper CT 30th Annual Meeting of the American-Society-of-Nephrology CY NOV 02-06, 1997 CL SAN ANTONIO, TEXAS SP Amer Soc Nephrol ID STAGE RENAL-DISEASE; BECK DEPRESSION INVENTORY; MAJOR DEPRESSION; MARITAL CONFLICT; BEHAVIORAL COMPLIANCE; SOCIAL RELATIONSHIPS; OPIOID-PEPTIDES; CHRONIC ILLNESS; SURVIVAL; CYTOKINES AB The effects of dyadic satisfaction and conflict have not been well defined in the hemodialysis (HD) population. The aim of this study was to determine whether the perception of decreased dyadic satisfaction was associated with mortality in patients treated with HD, and if so, whether there were different relationships between risk factors, and differential outcomes in men and women. A total of 174 HD patients, primarily African-Americans. involved in dyadic relationships for more than 6 mo had indices of dyadic satisfaction, depression, perception of illness effects, social support, behavioral compliance with the dialysis prescription, and plasma interleukin-1 (IL-1) and beta-endorphin levels measured. Cox proportional hazards models assessed relative mortality risks. Patients' dyadic satisfaction scores correlated with P-endorphin levels. There was no correlation of IL-1 or beta-endorphin with any psychosocial or behavioral compliance measure in the group as a whole. Correlations between psychosocial, medical, and neuroimmunologic variables were different in men and women. For women, dyadic satisfaction correlated with beta-endorphin levels, depression, and perception of illness. Women with higher dyadic satisfaction and decreased dyadic conflict were at decreased mortality risk, but dyadic adjustment indices were unassociated with differential survival in the larger group of men. Correlations between neuroendocrine and immune markers are different in African-American male and female HD patients. Greater dyadic satisfaction and lower dyadic conflict are independently associated with decreased mortality in female African-American HD patients, of the same order of magnitude as medical risk factors. Such effects may be attributable to a relationship between dyadic satisfaction and conflict and health-related behaviors, or through an effect on neuroendocrine or immunologic status. C1 George Washington Univ, Med Ctr, Dept Med,Sch Publ Hlth, Div Renal Dis & Hypertens, Washington, DC 20037 USA. George Washington Univ, Med Ctr, Dept Psychiat & Behav Sci, Sch Publ Hlth, Washington, DC 20037 USA. George Washington Univ, Dept Psychol, Washington, DC 20052 USA. Howard Univ, Med Ctr, Dept Human Dev & Psychoeduc Studies, Washington, DC 20059 USA. Howard Univ, Med Ctr, Dept Med, Washington, DC 20059 USA. NICHD, Sect Womens Hlth, Dev Endocrinol Branch, NIH, Bethesda, MD USA. Washington Hosp Ctr, Dept Med, Washington, DC 20010 USA. RP Kimmel, PL (reprint author), George Washington Univ, Med Ctr, Dept Med,Sch Publ Hlth, Div Renal Dis & Hypertens, 2150 Penn Ave NW, Washington, DC 20037 USA. FU NICHD NIH HHS [ZO1-HD-00641-01]; NIDDK NIH HHS [1-RO1-DK 45578] NR 53 TC 78 Z9 80 U1 1 U2 10 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 2000 VL 11 IS 8 BP 1518 EP 1525 PG 8 WC Urology & Nephrology SC Urology & Nephrology GA 337CY UT WOS:000088340900017 PM 10906166 ER PT J AU Gowdak, LHW Poliakova, L Li, ZH Grove, R Lakatta, EG Talan, M AF Gowdak, LHW Poliakova, L Li, ZH Grove, R Lakatta, EG Talan, M TI Induction of angiogenesis by cationic lipid-mediated VEGF(165) gene transfer in the rabbit ischemic hindlimb model SO JOURNAL OF VASCULAR SURGERY LA English DT Article; Proceedings Paper CT 2nd Annual Meeting of the American-Society-of-Gene-Therapy CY JUN 12, 1999 CL WASHINGTON, D.C. SP Amer Soc Gene Therapy ID ENDOTHELIAL GROWTH-FACTOR; IN-VIVO; LIMB ISCHEMIA; BLOOD-FLOW; DELIVERY; THERAPY; PHVEGF(165); EXPRESSION; DISEASE; VECTOR AB Purpose: The purpose of this study was to test the efficacy of a new cationic lipid formulation coupled with the cDNA encoding for the 165-residue form of vascular endothelial growth factor (VEGF(165)) to induce neovascularization and enhance blood flow in the rabbit ischemic hindlimb model. Methods: Two days after removal of their right femoral arteries, rabbits received intramuscular injections of different concentrations of VEGF(165) Or Saline solution in the ischemic thigh. Tissue perfusion and increased neovascularization of the ischemic limb were assessed weekly on the basis of the calf blood pressure ratio for the ischemic/nonischemic limbs, regional blood flow to the skeletal muscles as measured with radioactive microspheres, postmortem angiography, and histology. Results: At weeks 1 and 2 after surgery, animals treated with 1000 mu g of VEGP(165) had a 1.5-fold increase and a 2.5-fold increase, respectively, in the regional blood flow to both the adductor and gastrocnemius muscles of the ischemic limb. The blood pressure ratio was also greater in the treated animals than in the controls at weeks 2 and 3 after surgery. Early neovascularization in the VEGF(165) group was further documented at week 1 after surgery by more angiographically recognizable collateral vessels (angioscores were 64.13 +/- 2.51 and 38.28 +/- 3.82 for VEGF(165) and saline solution, respectively; P < .001) and by a threefold increase in the number of capillaries (vascular density) relative to the controls (P < .005). Conclusions: Intramuscular administration of a single dose of plasmid-liposomes encoding for VEGF(165) accelerates angiogenesis and increases blood flow in the rabbit hindlimb ischemic model. Therefore, this nonviral vector could be recommended for further testing for use in therapeutic angiogenesis. (J Vasc Surg 2000;32:343-52.). C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, Gene Therapy Unit,NIH, Baltimore, MD 21224 USA. Valentis Inc, Burlingame, CA USA. RP Talan, M (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, Gene Therapy Unit,NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Wolff Gowdak, Luis Henrique/C-4844-2012 OI Wolff Gowdak, Luis Henrique/0000-0002-6785-7506 NR 22 TC 26 Z9 28 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0741-5214 J9 J VASC SURG JI J. Vasc. Surg. PD AUG PY 2000 VL 32 IS 2 BP 343 EP 352 DI 10.1067/mva.2000.107566 PG 10 WC Surgery; Peripheral Vascular Disease SC Surgery; Cardiovascular System & Cardiology GA 343LH UT WOS:000088703500016 PM 10917995 ER PT J AU Van Berkel, V Barrett, J Tiffany, HL Fremont, DH Murphy, PM McFadden, G Speck, SH Virgin, HW AF Van Berkel, V Barrett, J Tiffany, HL Fremont, DH Murphy, PM McFadden, G Speck, SH Virgin, HW TI Identification of a gammaherpesvirus selective chemokine binding protein that inhibits chemokine action SO JOURNAL OF VIROLOGY LA English DT Article ID MURINE GAMMAHERPESVIRUS-68; MYXOMA VIRUS; BETA-CHEMOKINE; LATENT INFECTION; T1/35KDA FAMILY; GAMMA RECEPTOR; HOMOLOG; HERPESVIRUS; ENCODES; CYTOMEGALOVIRUS AB Chemokines are involved in recruitment and activation of hematopoietic cells at sites of infection and inflammation. The M3 gene of gamma HV68, a gamma-2 herpesvirus that infects and establishes a lifelong latent infection and chronic vasculitis in mice, encodes an abundant secreted protein during productive infection. The M3 gene is located in a region of the genome that is transcribed during latency. We report here that the M3 protein is a high affinity broad-spectrum chemokine scavenger, The M3 protein bound the CC chemokines human regulated upon activation of normal T-cell expressed and secreted (RANTES), murine macrophage inflammatory protein 1 alpha (MIP-1 alpha), and murine monocyte chemoattractant protein 1 (MCP-1), as well as the human CXC chemokine interleukin-8, the murine C chemokine lymphotactin, and the murine CX3C chemokine fractalkine with high affinity (K-d = 1.6 to 18.7 nM). M3 protein chemokine binding was selective, since the protein did not bind seven other CXC chemokines (K-d > 1 mu M). Furthermore, the M3 protein abolished calcium signaling in response to murine MIP-La and murine MCP-1 and not to murine KC or human stromal cell-derived factor 1 (SDF-1), consistent with the binding data. The M3 protein was also capable of blocking the function of human CC and CXC chemokines, indicating the potential for therapeutic applications, Since the M3 protein lacks homology to known chemokines, chemokine receptors, or chemokine binding proteins, these studies suggest a novel herpesvirus mechanism of immune evasion. C1 Washington Univ, Sch Med, Dept Pathol, St Louis, MO 63110 USA. Washington Univ, Sch Med, Ctr Immunol, St Louis, MO USA. Washington Univ, Sch Med, Dept Mol Microbiol & Immunol, St Louis, MO USA. Univ Western Ontario, Dept Microbiol & Immunol, London, ON, Canada. John P Robarts Res Inst, London, ON N6A 5K8, Canada. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Speck, SH (reprint author), Washington Univ, Sch Med, Dept Pathol, Box 8118,660 S Euclid Ave, St Louis, MO 63110 USA. FU NCI NIH HHS [R01 CA052004, CA43143, CA74730, R01 CA043143, R01 CA058524, R01 CA074730]; NIAID NIH HHS [AI39616]; NIGMS NIH HHS [T32 GM007200] NR 46 TC 143 Z9 146 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 2000 VL 74 IS 15 BP 6741 EP 6747 DI 10.1128/JVI.74.15.6741-6747.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900006 PM 10888612 ER PT J AU Durbin, AP Skiadopoulos, MH McAuliffe, JM Riggs, JM Surman, SR Collins, PL Murphy, BR AF Durbin, AP Skiadopoulos, MH McAuliffe, JM Riggs, JM Surman, SR Collins, PL Murphy, BR TI Human parainfluenza virus type 3 (PIV3) expressing the hemagglutinin protein of measles virus provides a potential method for immunization against measles virus and PIV3 in early infancy SO JOURNAL OF VIROLOGY LA English DT Article ID VACCINIA VIRUS; RHESUS-MONKEYS; ATTENUATION PHENOTYPES; RECOMBINANT VIRUSES; MATERNAL ANTIBODIES; 6-MONTH-OLD INFANTS; FUSION PROTEIN; REPLICATION; GLYCOPROTEINS; CANDIDATE AB Recombinant human parainfluenza virus type 3 (PIV3) was used as a vector to express the major protective antigen of measles virus, the hemagglutinin (HA) glycoprotein, in order to create a bivalent PIV3-measles virus that can be administered intranasally. The measles virus HA open reading frame (ORF) was inserted as an additional transcriptional unit into the N-P, P-M, or HA-neuraminidase (HN)-L gene junction of wild-type PIV3 or into the N-P or P-hl gene junction of an attenuated derivative of PIV3, termed rcp45L. The recombinant PIV3 (rPIV3) viruses bearing the IIA inserts replicated more slowly in vitro than their parental viruses but reached comparable peak titers of greater than or equal to 10(7.5) 50% tissue culture infective doses per mi. Each of the wild-type or cold-passaged 45L (cp35L) PIV3(IIA) chimeric viruses replicated 5- to 10-fold less well than its respective parent virus in the upper respiratory tract of hamsters, Thus, insertion of the similar to 2-kb ORF itself conferred attenuation, and this attenuation was additive to that conferred by the cp35L mutations. The attenuated cp45L PIV3(HA) recombinants induced a high level of resistance to replication of PIV3 challenge virus in hamsters and induced very high levels of measles virus neutralizing antibodies (>1:8,000) that are well in excess of those known to be protective in humans. rPIV3s expressing the HA gene in the N-P or P-hl junction induced about 400-fold more measles virus-neutralizing antibody than did the rPIV3 with the HA gene in the HN-L junction, indicating that the N-P or P-M junction appears to be the preferred insertion site. Previous studies indicated that the PIV3 cp45 virus, a more attenuated version of rcp45L, replicates efficiently in the respiratory tract of monkeys and is immunogenic and protective even when administered in the presence of very high titers of passively transferred PIV3 antibodies (A. P, Durbin, C. J, Cho, W, R Elkins, L, S, Wyatt, B, Moss, and B, R Murphy, J, Infect. Dis, 179:1345-1351, 1999). This suggests that this intranasally administered PIV3(HA) chimeric virus can be used to immunize infants with maternally acquired measles virus antibodies in whom the current parenterally administered live measles virus vaccine is ineffective. C1 NIAID, Infect Dis Lab, Resp Virus Sect, NIH, Bethesda, MD 20892 USA. RP Murphy, BR (reprint author), 7 Ctr Dr,MSC 0720, Bethesda, MD 20892 USA. NR 68 TC 42 Z9 43 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 2000 VL 74 IS 15 BP 6821 EP 6831 DI 10.1128/JVI.74.15.6821-6831.2000 PG 11 WC Virology SC Virology GA 333AE UT WOS:000088105900015 PM 10888621 ER PT J AU Taddesse-Heath, L Chattopadhyay, SK Dillehay, DL Lander, MR Nagashfar, Z Morse, HC Hartley, JW AF Taddesse-Heath, L Chattopadhyay, SK Dillehay, DL Lander, MR Nagashfar, Z Morse, HC Hartley, JW TI Lymphomas and high-level expression of murine leukemia viruses in CFW mice SO JOURNAL OF VIROLOGY LA English DT Article ID FOCUS-FORMING VIRUSES; SEQUENCES; GENOMES; DNA AB Historically, Swiss Webster mice of the CFW subline, both inbred and random-bred stocks, have been considered to have a low spontaneous occurrence of hematopoietic system tumors, and previous reports of infectious expression of murine leukemia viruses (MuLVs) have been rare and unremarkable, In marked contrast, in the present study of CFW mice from one source observed by two laboratories over a 2-year period, nearly 60% developed tumors, 85% of which were lymphomas, the majority of B-cell origin, All tumors tested expressed ecotropic MuLVs, and most expressed mink cell focus-inducing (MCF) MuLVs. Among normal mice of weanling to advanced age, over one-half were positive for ecotropic virus in tail or lymphoid tissues, and MCF virus was frequently present in lymphoid tissue, less often in tail. Patterns of ecotropic proviral integration indicated that natural infection occurred by both genetic and exogenous routes. Lymphomas were induced in NIH Swiss mice infected as neonates with tissue culture-propagated MuLVs isolated from normal and tumor tissue of CFW mice. C1 NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Dept Pathol, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Div Anim Resources, Atlanta, GA 30322 USA. RP Hartley, JW (reprint author), NIAID, Immunopathol Lab, NIH, 7 Ctr Dr,Room 7-304,MSC 0760, Bethesda, MD 20892 USA. OI Morse, Herbert/0000-0002-9331-3705 FU NIAID NIH HHS [N01-AI-45203] NR 28 TC 14 Z9 14 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 2000 VL 74 IS 15 BP 6832 EP 6837 DI 10.1128/JVI.74.15.6832-6837.2000 PG 6 WC Virology SC Virology GA 333AE UT WOS:000088105900016 PM 10888622 ER PT J AU Moriuchi, H Moriuchi, M Fauci, AS AF Moriuchi, H Moriuchi, M Fauci, AS TI Cathepsin G, a neutrophil-derived serine protease, increases susceptibility of macrophages to acute human immunodeficiency virus type 1 infection SO JOURNAL OF VIROLOGY LA English DT Article ID MONONUCLEAR-CELLS; HIV TRANSMISSION; IN-VITRO; REPLICATION; INHIBITOR; DISEASE; IDENTIFICATION; PROGRESSION; STIMULATION; ELASTASE AB Neutrophils dominate acute inflammatory responses that generally evolve into chronic inflammatory reactions mediated by monocyte/macrophages and lymphocytes. The latter cell types also serve as major targets for human immunodeficiency virus type 1 (HIV-1). In this study we have investigated the role of neutrophil products, particularly cathepsin G, in AN infection. Cathepsin G induced chemotaxis and production of proinflammatory cytokines by macrophages but not CD4(+) T cells. Pretreatment with cathepsin G markedly increased susceptibility of macrophages but not CD4(+) T cells to acute HIV-1 infection. When macrophages were exposed to pertussis toxin prior to cathepsin G treatment, the cathepsin G-mediated effect was almost abrogated, suggesting that enhancement of HIV-1 replication by cathepsin G requires Gi protein-mediated signal transduction, Although prolonged exposure to cathepsin G suppressed HIV infection of macrophages, serine protease inhibitors, which are exuded from the bloodstream later during inflammatory processes, neutralized the inhibitory effect. Neutrophil extracts or supernatants from neutrophil cultures, which contain cathepsin G, had effects similar to purified cathepsin G, Thus, cathepsin G, and possibly other neutrophil-derived serine proteases, may have multiple activities in HN-I infection of macrophages, including chemoattraction of monocyte/macrophage (HIV-1 targets) to inflamed tissue, activation of target cells, and increase in their susceptibility to acute HIV-1 infection. C1 Nagasaki Univ, Sch Med, Dept Pediat, Nagasaki 8528501, Japan. Nagasaki Univ, Grad Sch Med Sci, Dept Mol Microbiol & Immunol, Div Med Virol, Nagasaki 8528123, Japan. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Moriuchi, H (reprint author), Nagasaki Univ, Sch Med, Dept Pediat, 1-7-1 Sakamoto, Nagasaki 8528501, Japan. NR 29 TC 32 Z9 34 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 2000 VL 74 IS 15 BP 6849 EP 6855 DI 10.1128/JVI.74.15.6849-6855.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900018 PM 10888624 ER PT J AU Mahieux, R Pise-Masison, CA Lambert, PF Nicot, C De Marchis, L Gessain, A Green, P Hall, W Brady, JN AF Mahieux, R Pise-Masison, CA Lambert, PF Nicot, C De Marchis, L Gessain, A Green, P Hall, W Brady, JN TI Differences in the ability of human T-cell lymphotropic virus type 1 (HTLV-1) and HTLV-2 tax to inhibit p53 function SO JOURNAL OF VIROLOGY LA English DT Article ID TROPICAL SPASTIC PARAPARESIS; INFECTED CIRCULATING CELLS; BLOOD MONONUCLEAR-CELLS; WILD-TYPE P53; LEUKEMIA-VIRUS; TUMOR-SUPPRESSOR; TRANSCRIPTIONAL ACTIVATION; ABERRANT EXPRESSION; DRUG-USERS; SUBTYPE-B AB We have analyzed the functional activity of the p53 tumor suppressor in human T-cell lymphotropic virus type 2 (HTLV-2)-transformed cells. Abundant levels of the p53 protein mere detected in both HTLV-2A and -2B virus-infected cell lines. The p53 was functionally inactive, however, both in transient-transfection assays using a p53 reporter plasmid and in induction of p53-responsive genes in response to gamma irradiation. We further investigated HTLV-ZA Tax and HTLV-2B Tax effects on p53 activity, interestingly, although Tax-2A and -2B inactivate p53, the Tax-2A protein appears to inhibit p53 function less efficiently than either Tax-1 or Tax-2B. In transient-cotransfection assays, Tax-1 and Tax-2B inactivated p53 by 80%, while Tax2A reduced p53 activity by 20%. In addition, Tax-ZA does not increase the steady-state level of cellular p53 as well as Tax-1 or -2B does in the same assays. Cotransfection assays demonstrated that Tax-ZA could efficiently transactivate CREB-responsive promoters to the same level as Tax-1 and Tax-2B, indicating that the protein was functional. This report provides evidence of the first functional difference between the HTLV-ZA and -2B subtypes. This comparison of the action of HTLV-1 and HTLV-2 Tax proteins on p53 function will provide important insights into the mechanism of HTLV transformation. C1 NCI, Lab Receptor Biol & Gene Express, NIH, Bethesda, MD 20892 USA. NCI, Basic Res Lab, Sect Anim Models & Retroviral Infect, NIH, Bethesda, MD 20892 USA. NCI, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. Inst Pasteur, Unite Oncol Virale, F-75724 Paris, France. Ohio State Univ, Ctr Retrovirus Res, Dept Vet Biosci & Mol Virol, Columbus, OH 43210 USA. Ohio State Univ, Ctr Retrovirus Res, Dept Immunol, Columbus, OH 43210 USA. Ohio State Univ, Ctr Retrovirus Res, Dept Med Genet, Columbus, OH 43210 USA. Ohio State Univ, Ctr Comprehens Canc, Columbus, OH 43210 USA. Univ Coll Dublin, Dept Microbiol, Dublin 2, Ireland. RP Brady, JN (reprint author), NCI, Lab Receptor Biol & Gene Express, NIH, Bldg 41,B201, Bethesda, MD 20892 USA. NR 49 TC 40 Z9 40 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 2000 VL 74 IS 15 BP 6866 EP 6874 DI 10.1128/JVI.74.15.6866-6874.2000 PG 9 WC Virology SC Virology GA 333AE UT WOS:000088105900020 PM 10888626 ER PT J AU Endo, Y Igarashi, T Nishimura, Y Buckler, C Buckler-White, A Plishka, R Dimitrov, DS Martin, MA AF Endo, Y Igarashi, T Nishimura, Y Buckler, C Buckler-White, A Plishka, R Dimitrov, DS Martin, MA TI Short- and long-term clinical outcomes in rhesus monkeys inoculated with a highly pathogenic chimeric simian/human immunodeficiency virus SO JOURNAL OF VIROLOGY LA English DT Article ID BLOOD MONONUCLEAR-CELLS; PIG-TAILED MACAQUES; AIDS-LIKE DISEASE; CD4(+) T-CELLS; PRIMARY INFECTION; NEUTRALIZING ANTIBODIES; NATURAL-HISTORY; HIV-1 INFECTION; VIRAL DYNAMICS; TYPE-1 AB A highly pathogenic simian/human immunodeficiency virus (SHIV), SHIVDH12R, isolated from a rhesus macaque that had been treated with anti-human CD8 monoclonal antibody at the time of primary infection with the nonpathogenic, molecularly cloned SHIVDH12, induced marked and rapid CD4(+) T cell loss in all rhesus macaques intravenously inoculated with 1.0 50% tissue culture infective dose (TCID50) to 4.1 x 10(5) TCID(50)s of virus. Animals inoculated with 650 TCID(50)s of SHIVDH12R or more experienced irreversible CD4(+) T lymphocyte depletion and developed clinical disease requiring euthanasia between weeks 12 and 23 postinfection, In contrast, the CD4(+) T-cell numbers in four of five monkeys receiving 25 TCID(50)s of SHIVDH12R or less stabilized at low levels, and these surviving animals produced antibodies capable of neutralizing SHIVDH12R. In the fifth monkey, no recovery from the CD4(+) T cell decline occurred, and the animal had to be euthanized. Viral RNA levels, subsequent to the initial peak of infection but not at peak viremia, correlated with the virus inoculum size and the eventual clinical course. Both initial infection rate constants, k, and decay constants, d, were determined, but only the latter were statistically correlated to clinical outcome. The attenuating effects of reduced inoculum size were also observed when virus was inoculated by the mucosal route. Because the uncloned SHIVDH12R stock possessed the genetic properties of a lentivirus quasispecies, we were able to assess the evolution of the input virus swarm in animals surviving the acute infection by monitoring the emergence of neutralization escape viral variants. C1 NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. NCI, Lab Expt & Computat Biol, NIH, Frederick, MD 21702 USA. RP Martin, MA (reprint author), NIAID, Mol Microbiol Lab, NIH, Bldg 4,Rm 315,4 Ctr Dr,MSC 0460, Bethesda, MD 20892 USA. NR 41 TC 60 Z9 63 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 2000 VL 74 IS 15 BP 6935 EP 6945 DI 10.1128/JVI.74.15.6935-6945.2000 PG 11 WC Virology SC Virology GA 333AE UT WOS:000088105900026 PM 10888632 ER PT J AU Lee, S Tiffany, HL King, L Murphy, PM Golding, H Zaitseva, MB AF Lee, S Tiffany, HL King, L Murphy, PM Golding, H Zaitseva, MB TI CCR8 on human thymocytes functions as a human immunodeficiency virus type 1 coreceptor SO JOURNAL OF VIROLOGY LA English DT Article ID CHEMOKINE RECEPTOR CCR8; HUMAN THYMIC IMPLANTS; CELLS IN-VITRO; SCID-HU MICE; HIV-1 INFECTION; DIFFERENTIAL TROPISM; FUSION COFACTOR; GP120 BINDING; CXCR4; VIVO AB To determine whether human immunodeficiency virus type 1 (HIV-1) coreceptors besides CXCR4 and CCR5 are involved in HIV-1 infection of the thymus, we focused on CCR8, a receptor for the chemokine I-309, because of its high expression in the thymus. Similar levels of CCR8 mRNA were detected in immature and mature primary human thymocytes. Consistent with this, [I-125]I-309 was shown to bind specifically and with similar affinity to the surface of immature and mature human thymocytes. Fusion of human thymocytes with cells expressing HIV-1 X4 or X 4R5 envelope glycoprotein was inhibited by I-309 in a dose-dependent manner. In addition, I-309 partially inhibited productive infection of human thymocytes by X4, R5, and X4R5 HIV-1 strains. Our data provide the first evidence that CCR8 functions as an HIV-1 coreceptor on primary human cells and suggest that CCR8 may contribute to HIV-l-induced thymic pathogenesis. C1 CBER, FDA, Div Viral Prod, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Zaitseva, MB (reprint author), CBER, FDA, Div Viral Prod, Bldg 29B,8800 Rockville Pike, Bethesda, MD 20892 USA. NR 46 TC 36 Z9 36 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 2000 VL 74 IS 15 BP 6946 EP 6952 DI 10.1128/JVI.74.15.6946-6952.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900027 PM 10888633 ER PT J AU Anderson, JA Pathak, VK Hu, WS AF Anderson, JA Pathak, VK Hu, WS TI Effect of the murine leukemia virus extended packaging signal on the rates and locations of retroviral recombination SO JOURNAL OF VIROLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; HIGH NEGATIVE INTERFERENCE; 5' NONTRANSLATED REGION; GENETIC-RECOMBINATION; HOMOLOGOUS RECOMBINATION; REVERSE TRANSCRIPTION; STRAND TRANSFERS; INTERNAL ENTRY; SARCOMA-VIRUS; TUMOR-VIRUSES AB Reverse transcriptase (RT) switches templates frequently during DNA synthesis; the acceptor template can be the same RNA (intramolecular) or the copackaged RNA (intermolecular). Previous results indicated that intramolecular template switching occurred far more frequently than intermolecular template switching. We hypothesized that intermolecular template-switching events (recombination) occurred at a lower efficiency because the copackaged RNA was not accessible to the RT. To test our hypothesis, the murine leukemia virus (MLV) extended packaging signal (Psi(+)) containing a dimer linkage structure (DLS) was relocated from the 5' untranslated region (UTR) to between selectable markers, allowing the two viral RNAs to interact closely in this region. It was found that the overall maximum recombination rates of vectors with yr in the 5' UTR or Psi(+) between selectable markers were not drastically different. However, vectors with Psi(+) located between selectable markers reached a plateau of recombination rate at a shorter distance. This suggested a limited enhancement of recombination by Psi(+). The locations of the recombination events were also examined by using restriction enzyme markers. Recombination occurred in all four regions between the selectable markers; the region containing 5' Psi(+) including DLS did not undergo more recombination than expected from the size of the region. These experiments indicated that although the accessibility of the copackaged RNA was important in recombination, other factors existed to limit the number of viruses that were capable of undergoing intermolecular template switching. In addition, recombinants with multiple template switches were observed at a frequency much higher than expected, indicating the presence of high negative interference in the MLV-based system. This extends our observation with the spleen necrosis virus system and suggests that high negative interference may he a common phenomenon in retroviral recombination. C1 NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Frederick, MD 21702 USA. W Virginia Univ, Sch Med, Dept Microbiol & Immunol, Morgantown, WV 26506 USA. W Virginia Univ, Sch Med, Dept Biochem, Morgantown, WV 26506 USA. W Virginia Univ, Sch Med, Mary Babb Randolph Canc Ctr, Morgantown, WV 26506 USA. RP Hu, WS (reprint author), NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Bldg 535,Room 336, Frederick, MD 21702 USA. FU NCI NIH HHS [CA58875] NR 51 TC 15 Z9 15 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 2000 VL 74 IS 15 BP 6953 EP 6963 DI 10.1128/JVI.74.15.6953-6963.2000 PG 11 WC Virology SC Virology GA 333AE UT WOS:000088105900028 PM 10888634 ER PT J AU Peacock, CD Lin, MY Ortaldo, JR Welsh, RM AF Peacock, CD Lin, MY Ortaldo, JR Welsh, RM TI The virus-specific and allospecific cytotoxic T-lymphocyte response to lymphocytic choriomeningitis virus is modified in a subpopulation of CD8(+) T cells coexpressing the inhibitory major histocompatibility complex class I receptor Ly49G2 SO JOURNAL OF VIROLOGY LA English DT Article ID MHC CLASS-I; NATURAL-KILLER-CELLS; INFECTIOUS-MONONUCLEOSIS; CYTO-TOXICITY; EXPRESSION; MOLECULES; MICE; RECOGNITION; BINDING; FAMILY AB The role of negatively signaling NK cell receptors of the Ly49 family on the specificity of the acute CD8(+) cytotoxic T-lymphocyte (CTL) response was investigated in lymphocytic choriomeningitis virus (LCMV)infected C57BL/6 mice. Activated CD8(+) T cells coexpressing Ly49G2 expanded during LCMV infection, and T-cell receptor analyses by flow cytometry and CDR3 spectratyping revealed a unique polyclonal T-cell population in the Ly49G2(+) fraction. These cells lysed syngeneic targets infected,with LCMV or coated with two of three LCMV immunodominant peptides examined. Transfection of these sensitive targets with H2D(d), a ligand for Ly49G2, inhibited lysis, This was reversed by antibody to Ly49G2, indicating effective negative signaling. LCMV characteristically induces an anti-H2(d) allospecific T-cell response that includes T-cell clones cross-reactive between allogeneic and LCMV-infected syngeneic targets. The CD8(+) Ly49G2(+) population mediated no allospecific killing, nor was any NK-like killing observed against YAC-1 cells. This study shows that CD8+ Ly49G2(+) cells participate in the virus-induced CTL response but lyse a more restricted range of targets than the rest of the virus-induced CTL population. C1 Univ Massachusetts, Med Ctr, Dept Pathol, Worcester, MA 01655 USA. NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Expt Immunol Lab, Frederick, MD 21702 USA. RP Welsh, RM (reprint author), Univ Massachusetts, Med Ctr, Dept Pathol, 55 Lake Ave N, Worcester, MA 01655 USA. FU NCI NIH HHS [CA34461, R01 CA034461] NR 32 TC 29 Z9 29 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 2000 VL 74 IS 15 BP 7032 EP 7038 DI 10.1128/JVI.74.15.7032-7038.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900036 PM 10888642 ER PT J AU Bukreyev, A Whitehead, SS Prussin, C Murphy, BR Collins, PL AF Bukreyev, A Whitehead, SS Prussin, C Murphy, BR Collins, PL TI Effect of coexpression of interleukin-2 by recombinant respiratory syncytial virus on virus replication, immunogenicity, and production of other cytokines SO JOURNAL OF VIROLOGY LA English DT Article ID ENCODING MURINE IL-2; IMMUNODEFICIENT MICE; INTERFERON-GAMMA; IMMUNE-RESPONSES; MESSENGER-RNA; FOREIGN GENE; INFECTION; EXPRESSION; RECOVERY; VACCINES AB We constructed rRSV/mIL-2, a recombinant respiratory syncytial virus (rRSV) containing the coding sequence of murine interleukin-2 (mIL-2) in a transcription cassette inserted into the G-F intergenic region, The recovered virus (rRSV/mIL-2) expressed high levels (up to 2.8 mu g/ml) of mIL-2 in cell culture. Replication of rRSV/mIL-2 in vitro was reduced up to 13.6-fold from that of wild-type (wt) rRSV, an effect that was due to the presence of the foreign insert but was not specific to mIL-2, Replication of the rRSV/mIL-2 virus in the upper and lower respiratory tracts of BALB/c mice was reduced up to 6.3-fold, an effect that was specific to mIL-2, The antibody response, including the levels of RSV-specific serum immunoglobulin G1 (IgG1), IgG2a, IgA, and total IgG, and the level of protective efficacy against wt RSV challenge were not significantly different from those of wt rRSV, Analysis of total pulmonary cytokine mRNA isolated 1 and 4 days following infection with rRSV/mIL-2 revealed elevated levels of mRNA for IL-2, gamma interferon (IFN-gamma), IL-4, IL-5, IL-6, IL-10, IL-13, and IL-12 p40 compared to those for,vt rRSV. Flow cytometry of total pulmonary mononuclear cells isolated 10 days following infection with rRSV/mIL-2 revealed increased levels of CD4(+) T lymphocytes expressing either IFN-gamma or IL-4 compared to those of wt rRSV. These elevations in cytokine mRNA or cytokine-expressing CD4(+) cells relative to those of wt rRSV-primed animals were not observed following challenge with wt RSV on day 28, Thus, the expression of mIL-2 by rRSV was associated with a modest attenuation of virus growth in vivo, induction of serum antibodies at levels comparable to that of wt rRSV, and transient increases in both the Th1 and Th2 CD4(+) lymphocytes and cytokine mRNAs compared to those of wt rRSV. C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. NIAID, Lab Allerg Dis, Bethesda, MD 20892 USA. RP Collins, PL (reprint author), NIAID, Infect Dis Lab, NIH, Bldg 7,Room 100,7 Ctr Dr,MSC 0720, Bethesda, MD 20892 USA. OI Prussin, Calman/0000-0002-3917-3326 NR 26 TC 23 Z9 26 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 2000 VL 74 IS 15 BP 7151 EP 7157 DI 10.1128/JVI.74.15.7151-7157.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900050 PM 10888656 ER PT J AU Glamann, J Hirsch, VM AF Glamann, J Hirsch, VM TI Characterization of a macaque recombinant monoclonal antibody that binds to a CD4-induced epitope and neutralizes simian immunodeficiency virus SO JOURNAL OF VIROLOGY LA English DT Article ID PBL-SCID MICE; PASSIVE-IMMUNIZATION; SYNERGISTIC NEUTRALIZATION; ENVELOPE GLYCOPROTEIN; HIV TYPE-1; INFECTION; GP120; SIV; CYNOMOLGUS; RESISTANT AB A potent neutralizing Fab fragment from a long-term survivor of simian immunodeficiency virus (SIVsm) infection was used to construct a recombinant macaque immunoglobulin G1 kappa (IgG1 kappa) molecule, designated IgG1-201. A Chinese hamster ovary cell line expressing IgG1-201 was derived by stable transfection and optimized for antibody secretion by methotrexate selection and dihydrofolate reductase gene amplification. IgG1-201 effectively neutralized the homologous, molecularly cloned SIVsmH4 virus but had no activity against the heterologous SIVmac251/BK28 virus. The previously characterized, neutralization-resistant SIVsmE543-3 virus was also not neutralized by IgG1-201. Binding to SIVsmH4 gp120 was enhanced in the presence of recombinant soluble CD4, suggesting that IgG1-201 bound a CD4-induced epitope. IgG1-201 immunoprecipitated the SIVsmH4 but not the SIVsmE543-3 envelope despite a close relationship between these two clones. Immunoprecipitation of a panel of SIVsmH4/SIVsmE543-3 chimeric viruses tentatively assigned the neutralization epitope to the third constant domain, immediately C terminal to the V3 loop. These findings suggest the presence of at least one CD4-induced neutralization epitope on SIV as is the case with human immunodeficiency virus type 1. C1 NIAID Twinbrook II Facil, Mol Microbiol Lab, NIH, Rockville, MD 20852 USA. RP Hirsch, VM (reprint author), NIAID Twinbrook II Facil, Mol Microbiol Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. NR 44 TC 8 Z9 8 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 2000 VL 74 IS 15 BP 7158 EP 7163 DI 10.1128/JVI.74.15.7158-7163.2000 PG 6 WC Virology SC Virology GA 333AE UT WOS:000088105900051 PM 10888657 ER PT J AU Haag, AL Lin, JH Levin, HL AF Haag, AL Lin, JH Levin, HL TI Evidence for the packaging of multiple copies of Tfl mRNA into particles and the trans priming of reverse transcription SO JOURNAL OF VIROLOGY LA English DT Article ID METHIONINE TRANSFER-RNA; SCHIZOSACCHAROMYCES-POMBE; FISSION YEAST; STEM-LOOP; MECHANISM; DNA; REQUIRES; PRIMER AB Long terminal repeat (LTR)-containing retrotransposons and retroviruses are close relatives that possess similar mechanisms of reverse transcription. The particles of retroviruses package hco copies of viral mRNA that both function as templates for the reverse transcription of the element. We studied the LTR-retrotransposon Tf1 of Schizosaccharomyces pombe to test whether multiple copies of transposon mRNA participate In the production of cDNA. Using the unique self-priming property of Tf1, we obtained evidence that multiple copies of Tf1 mRNA mere packaged into virus-like particles. By coexpressing two distinct versions of Tf1, we found that the bulk of reverse transcription that was initiated on one mRNA template was subsequently transferred to others. In addition, the first 11 nucleotides of one mRNA were able to prime, in trans, the reverse transcription of another mRNA. C1 NICHHD, Lab Eukaryot Gene Regulat, NIH, Bethesda, MD 20892 USA. RP Levin, HL (reprint author), NICHHD, Lab Eukaryot Gene Regulat, NIH, Bethesda, MD 20892 USA. NR 18 TC 5 Z9 5 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 2000 VL 74 IS 15 BP 7164 EP 7170 DI 10.1128/JVI.74.15.7164-7170.2000 PG 7 WC Virology SC Virology GA 333AE UT WOS:000088105900052 PM 10888658 ER PT J AU Svarovskaia, ES Delviks, KA Hwang, CK Pathak, VK AF Svarovskaia, ES Delviks, KA Hwang, CK Pathak, VK TI Structural determinants of murine leukemia virus reverse transcriptase that affect the frequency of template switching SO JOURNAL OF VIROLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; RNA SECONDARY STRUCTURE; ACTIVATING RETROVIRAL VECTORS; ALANINE-SCANNING MUTAGENESIS; SINGLE REPLICATION CYCLE; 5' NONTRANSLATED REGION; DNA-SYNTHESIS; STRAND TRANSFER; NUCLEOCAPSID PROTEIN; SARCOMA VIRUS AB Retroviral reverse transcriptases (RTs) frequently switch templates within the same RNA or between copackaged viral RNAs to generate mutations acid recombination. To identify structural elements of murine leukemia virus RT important for template switching, we developed an in vivo assay in which BT template switching within direct repeats functionally reconstituted the green fluorescent protein gene. We quantified the effect of mutations in the YXDD motif, the deoxynucleoside triphosphate binding site, the thumb domain, and the RNase II domain of RT and hydroxyurea treatment on the frequencies of template switching. Hydroxyurea treatment and some mutations in RT increased the frequency of RT template switching up to fivefold, while all of the mutations tested in the RNase Il domain decreased the frequency of template switching by twofold. Based on these results, we propose a dynamic copy choice model in which both the rate of DNA polymerization and the rate of RNA degradation influence the frequency of RT template switching. C1 NCI, HIV Drug Resistance Program, FCRDC, Frederick, MD 21702 USA. W Virginia Univ, Dept Biochem, Morgantown, WV 26506 USA. W Virginia Univ, Dept Microbiol & Immunol, Morgantown, WV 26506 USA. RP Pathak, VK (reprint author), NCI, HIV Drug Resistance Program, FCRDC, Bldg 535,Rm 334, Frederick, MD 21702 USA. RI Delviks-Frankenberry, Krista/M-4822-2013 FU NCI NIH HHS [CA58875] NR 67 TC 60 Z9 62 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 2000 VL 74 IS 15 BP 7171 EP 7178 DI 10.1128/JVI.74.15.7171-7178.2000 PG 8 WC Virology SC Virology GA 333AE UT WOS:000088105900053 PM 10888659 ER PT J AU Miller, RJ Cairns, JS Bridges, S Sarver, N AF Miller, RJ Cairns, JS Bridges, S Sarver, N TI Human immunodeficiency virus and AIDS: Insights from animal lentiviruses SO JOURNAL OF VIROLOGY LA English DT Review ID INFECTIOUS-ANEMIA VIRUS; PROTEIN ZINC FINGERS; CELL-CYCLE ARREST; VIF PROTEIN; IN-VITRO; SOOTY MANGABEYS; VPR PROTEIN; TAT PROTEIN; HIV-1 TAT; TYPE-1 C1 NIAID, Div Aids, Targeted Intervent Branch, NIH, Bethesda, MD 20892 USA. RP Sarver, N (reprint author), NIAID, Div Aids, Targeted Intervent Branch, NIH, 6700-B Rockledge Dr,Room 4126, Bethesda, MD 20892 USA. NR 54 TC 56 Z9 57 U1 0 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 2000 VL 74 IS 16 BP 7187 EP 7195 DI 10.1128/JVI.74.16.7187-7195.2000 PG 9 WC Virology SC Virology GA 337DP UT WOS:000088342400001 PM 10906172 ER PT J AU Yuan, B Campbell, S Bacharach, E Rein, A Goff, SP AF Yuan, B Campbell, S Bacharach, E Rein, A Goff, SP TI Infectivity of moloney murine leukemia virus defective in late assembly events is restored by late assembly domains of other retroviruses SO JOURNAL OF VIROLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; ROUS-SARCOMA VIRUS; PFIZER MONKEY VIRUS; GAG PROTEIN; IN-VITRO; CYTOPLASMIC DOMAIN; ZINC-FINGER; MUTATIONS; TYPE-1; MUTANTS AB The p12 region of the Moloney murine leukemia virus (M-MuLV) Gag protein contains a PPPY motif important for efficient virion assembly and release. To probe the function of the PPPY motif, a series of insertions of homologous and heterologous motifs from other retroviruses were introduced at various positions in a mutant gag gene Lacking the PPPY motif. The assembly defects of the PPPY deletion mutant could be rescued by insertion of a wild-type PPPY motif and flanking sequences at several ectopic positions in the Gag protein. The late assembly domain (L-domain) of Rous sarcoma virus (RSV) or human immunodeficiency virus type 1 (HIV-1) could also fully or partially restore M-MuLV assembly when introduced into matrix, p12, or nocleocapsid domains of the mutant M-MuLV Gag protein lacking the PPPY motif. Strikingly, mutant viruses carrying the RSV or the HIV-1 L-domain at the original location of the deleted PPPY motif were replication competent in rodent cells. These data suggest that the PPPY motif of M-MuLV acts in a partially position-independent manner and is functionally interchangeable with L-domains of other retroviruses. Electron microscopy studies revealed that deletion of the entire p12 region resulted in the formation of tube-like rather than spherical particles. Remarkably, the PPPY deletion mutant formed chain structures composed of multiple viral particles linked on the cell surface. Many of the mutants with heterologous L-domains released virions with wild-type morphology. C1 Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA. Columbia Univ Coll Phys & Surg, Integrated Program Cellular Mol & Biophys Studies, New York, NY 10032 USA. NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Frederick, MD 21702 USA. RP Yuan, B (reprint author), Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, 701 W 168th St, New York, NY 10032 USA. RI Goff, Stephen/K-6337-2014 OI Goff, Stephen/0000-0003-0693-5547 FU NCI NIH HHS [R01 CA030488, CA 30488, R37 CA030488] NR 38 TC 130 Z9 130 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 2000 VL 74 IS 16 BP 7250 EP 7260 DI 10.1128/JVI.74.16.7250-7260.2000 PG 11 WC Virology SC Virology GA 337DP UT WOS:000088342400008 PM 10906179 ER PT J AU De La Fuente, C Santiago, F Chong, SY Deng, LW Mayhood, T Fu, P Stein, D Denny, T Coffman, F Azimi, N Mahieux, R Kashanchi, F AF De La Fuente, C Santiago, F Chong, SY Deng, LW Mayhood, T Fu, P Stein, D Denny, T Coffman, F Azimi, N Mahieux, R Kashanchi, F TI Overexpression of p21(waf1) in human T-cell lymphotropic virus type 1-infected cells and its association with cyclin A/cdk2 SO JOURNAL OF VIROLOGY LA English DT Article ID KINASE INHIBITOR P21; TAX PROTEIN; HTLV-I; RETINOBLASTOMA PROTEIN; DEPENDENT KINASES; DNA-BINDING; C-ABL; EXPRESSION; PHOSPHORYLATION; P53 AB Human T-cell lymphotropic virus type 1 (HTLV-1) is associated with adult T-cell leukemia (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). T-cell transformation is mainly due to the actions of the viral phosphoprotein Tax. Tax interacts with multiple transcriptional factors, aiding the transcription of many cellular genes. Here, we report that the cyclin-dependent kinase inhibitor p21/waf1 is overexpressed in all HTLV-1-infected cell lines tested as well as in ATL and HAM/TSP patient samples. Tax was found to be able to transactivate the endogenous p21/waf1 promoter, as detected by RNase protection, as well as activate a series of wild-type and 5'-deletion constructs linked to a luciferase reporter cassette. Wild-type but not a mutant form of Tax (M47) transactivated the p21/waf1 promoter in a p53-independent manner and utilized a minimal promoter that contained E2A and TATA box sequences. The p21/waf1 protein was reproducibly observed to be complexed with cyclin A/cdk2 and not with any other known G(1), S, or G(2)/M cyclins. Functionally, the association of p21/cyclin A/cdk2 decreased histone H1 phosphorylation in vitro, as observed in immunoprecipitations followed by kinase assays, and affected other substrates, such as the C terminus of Rb protein involved in c-Ab1 and histone deacetylase-1 (HDAC1) regulation. Interestingly, upon the use of a stress signal, such as gamma-irradiation, we found that the p21/cyclin A/cdk2 complex was able to block all known phosphorylation sites on the Rb molecule. Finally, using elutriated cell cycle fractions and a stress signal, we observed that the HTLV-1-infected T cells containing wild-type Tax, which had been in early or mid-G(1) phase prior to gamma-irradiation, arrested in G(1) and did not undergo apoptosis. This may be an important mechanism for an oncogenic virus such as HTLV-1 to stop the host at the G(1)/S boundary and to repair the damaged DNA upon injury, prior to S-phase entry. C1 Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Biochem & Mol Biol, Newark, NJ 07103 USA. Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Pediat, Newark, NJ 07103 USA. Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Pathol, Newark, NJ 07103 USA. NCI, NIH, Bethesda, MD 20874 USA. Inst Pasteur, Dept SIDA Retrovirus, Unite Oncol Virale, F-75724 Paris, France. RP Kashanchi, F (reprint author), Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Biochem & Mol Biol, MSB E635, Newark, NJ 07103 USA. OI Denny, Thomas/0000-0002-7364-8276 FU NIAID NIH HHS [AI13969, AI43894, R01 AI043894, AI42524] NR 35 TC 65 Z9 65 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 2000 VL 74 IS 16 BP 7270 EP 7283 DI 10.1128/JVI.74.16.7270-7283.2000 PG 14 WC Virology SC Virology GA 337DP UT WOS:000088342400010 PM 10906181 ER PT J AU Azimi, N Tagaya, Y Mariner, J Waldmann, TA AF Azimi, N Tagaya, Y Mariner, J Waldmann, TA TI Viral activation of interleukin-15 (IL-15): Characterization of a virus-inducible element in the IL-15 promoter region SO JOURNAL OF VIROLOGY LA English DT Article ID NF-KAPPA-B; INTERFERON REGULATORY FACTOR-3; IFN-BETA-GENE; NATURAL-KILLER-CELLS; TRANSCRIPTIONAL ACTIVATOR; MESSENGER-RNA; T-CELLS; IN-VIVO; EXPRESSION; INDUCTION AB We identified an interferon regulatory factor motif (IRF-E) upstream of an NF-kappa B binding site in the interleukin-15 (IL-15) promoter. Since these two motifs are part of the virus-inducible enhancer region of the beta interferon promoter, we speculated that there might be similar responses of these two genes to stimuli such as viruses. To test this hypothesis, L929 cells were infected with Newcastle disease virus (NDV), which led to the induction of IL-15 mRNA and protein expression. Using IL-15 promoter-reporter deletion constructs, a virus-inducible region, encompassing IRF-E, NF-kappa B, and a 13-nucleotide sequence flanked by these two motifs, was mapped to the -295-to--233 position relative to the transcription initiation site. Using cotransfection studies, it was demonstrated that all three motifs were essential to achieve the maximum promoter activity induced by IRF-1 and NF-kappa B expression plasmids. The presence of a virus-inducible region in the IL-15 promoter suggests a role for IL-15 as a component of host antiviral defense mechanisms. C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Azimi, N (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, Bldg 10,Rm 4N-102,10 Ctr Dr MSC 1374, Bethesda, MD 20892 USA. NR 43 TC 52 Z9 53 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 2000 VL 74 IS 16 BP 7338 EP 7348 DI 10.1128/JVI.74.16.7338-7348.2000 PG 11 WC Virology SC Virology GA 337DP UT WOS:000088342400016 PM 10906187 ER PT J AU da Fonseca, FG Wolffe, EJ Weisberg, A Moss, B AF da Fonseca, FG Wolffe, EJ Weisberg, A Moss, B TI Characterization of the vaccinia virus H3L envelope protein: Topology and posttranslational membrane insertion via the C-terminal hydrophobic tail SO JOURNAL OF VIROLOGY LA English DT Article ID INTERMEDIATE COMPARTMENT; ENDOPLASMIC-RETICULUM; IDENTIFICATION; GENE; CONTAIN; GENOME; FUSION; FORMS; A27L AB The vaccinia virus H3L open reading frame encodes a 324-amino-acid immunodominant membrane component of virus particles. Biochemical and microscopic studies demonstrated that the H3L protein was expressed late in infection, accumulated in the cytoplasmic viral Factory regions, and associated primarily with amorphous material near immature virions and with intracellular virion membranes. Localization of the H3L protein on the surfaces of viral particles and anchorage via the hydrophobic tail were consistent with its extraction by NP-40 in the absence of reducing agents, its trypsin sensitivity, its reactivity with a membrane-impermeable biotinylation reagent, and its immunogold labeling with an antibody to a peptide comprising amino acids 247 to 259, The H3L protein, synthesized in a coupled in vitro transcription/translation system, was tightly anchored to membranes as determined by resistance to Na2CO3 (pH 11) extraction and cytoplasmically oriented as shown by sensitivity to proteinase K digestion. Further studies demonstrated that membrane insertion of the H3L protein occurred posttranslationally and that the C-terminal hydrophobic domain was necessary and sufficient for this to occur. These data indicated that the H3L protein is a member of the C-terminal anchor family and supported a model in which it is synthesized on free ribosomes and inserts into the membranes of viral particles during their maturation. 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 37 TC 61 Z9 63 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 2000 VL 74 IS 16 BP 7508 EP 7517 DI 10.1128/JVI.74.16.7508-7517.2000 PG 10 WC Virology SC Virology GA 337DP UT WOS:000088342400033 PM 10906204 ER PT J AU da Fonseca, FG Wolffe, EJ Weisberg, A Moss, B AF da Fonseca, FG Wolffe, EJ Weisberg, A Moss, B TI Effects of deletion or stringent repression of the H3L envelope gene on vaccinia virus replication SO JOURNAL OF VIROLOGY LA English DT Article ID ACTIN-CONTAINING MICROVILLI; OPEN READING FRAME; TO-CELL SPREAD; VIRION MORPHOGENESIS; EXTRACELLULAR VIRUS; TAIL FORMATION; PROTEIN; MEMBRANE; IDENTIFICATION; DISSEMINATION AB The C-terminal membrane anchor protein encoded by the H3L open reading frame of vaccinia virus is located on the surfaces of intracellular mature virions. To investigate the role of the H3L protein, we constructed deletion (vH3 Delta) and inducible (vH3i) null mutants. The H3L protein was not detected in lysates of cells infected with vH3 Delta or vH3i in the absence of inducer. Under these conditions, plaques were small and round instead of large and comet shaped, indicative of decreased virus replication or cell-to-cell spread. The mutant phenotype was correlated with reduced yields of infectious intra- and extracellular virus in one-step growth experiments. The defect in vH3i replication could not be attributed to a role of the H3L protein in virus binding, internalization, or any event prior to late gene expression. Electron microscopic examination of cells infected with vH3 Delta or vH3i in the absence of inducer revealed that virion assembly was impaired, resulting in a high ratio of immature to mature virus forms with an accumulation of crescent membranes adjacent to granular material and DNA crystalloids, The absence of the H3L protein did not impair the membrane localization of virion surface proteins encoded by the A27L, D8L, and L1R genes. The wrapping of virions and actin tail formation were not specifically blocked, but there was an apparent defect in low-pN-mediated syncytium formation that could be attributed to decreased virus particle production. The phenotypes of the H3L deletion and repression mutants were identical to each other but differed from those produced by null mutations of genes encoding other vaccinia virus membrane components. 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 49 TC 27 Z9 29 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 2000 VL 74 IS 16 BP 7518 EP 7528 DI 10.1128/JVI.74.16.7518-7528.2000 PG 11 WC Virology SC Virology GA 337DP UT WOS:000088342400034 PM 10906205 ER PT J AU Johnson, BR Jang, SJ De Li, S Larson, D AF Johnson, BR Jang, SJ De Li, S Larson, D TI The 'invisible institution' and black youth crime: The church as an agency of local social control SO JOURNAL OF YOUTH AND ADOLESCENCE LA English DT Article ID DELINQUENCY; RELIGIOSITY; PARTICIPATION; NEIGHBORHOOD; HELLFIRE; CONTEXT; IMPACT; FEAR AB We examine the degree to which an individual's religious involvement significantly mediates and buffers the effects of neighborhood disorder on youth crime. To test this hypothesis, we analyzed the fifth wave of data from the National Youth Survey, focusing on black respondents given the historical as well as contemporary significance of the African-American church for black Americans. Results front estimating a series of regression models indicated that (1) the effects of neighborhood disorder on crime among black youth are partly mediated by an individual's religious involvement (measured by the frequency of attending religious services) and (2) involvement of African-American youth in religious institutions significantly buffers or interacts with the effects of neighborhood disorder on crime and, in particular serious crime. Theoretical and methodological implications of the present findings are briefly discussed. C1 Louisiana State Univ, Dept Sociol, Baton Rouge, LA 70803 USA. Univ Maryland, Dept Criminol & Criminal Justice, College Pk, MD 20742 USA. Natl Inst Healthcare Res, Rockville, MD USA. Univ Penn, Philadelphia, PA 19104 USA. Duke Univ, Durham, NC 27706 USA. NIH, Bethesda, MD 20892 USA. RP Johnson, BR (reprint author), Louisiana State Univ, Dept Sociol, Baton Rouge, LA 70803 USA. RI Jang, Sung Joon/B-7105-2009 OI Jang, Sung Joon/0000-0003-2228-158X NR 53 TC 49 Z9 49 U1 4 U2 8 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0047-2891 J9 J YOUTH ADOLESCENCE JI J. Youth Adolesc. PD AUG PY 2000 VL 29 IS 4 BP 479 EP 498 DI 10.1023/A:1005114610839 PG 20 WC Psychology, Developmental SC Psychology GA 345LZ UT WOS:000088818600006 ER PT J AU Moscrip, TD Ingram, DK Lane, MA Roth, GS Weed, JL AF Moscrip, TD Ingram, DK Lane, MA Roth, GS Weed, JL TI Locomotor activity in female rhesus monkeys: Assessment of age and calorie restriction effects SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID TERM DIETARY RESTRICTION; FISCHER-344 RATS; BLOOD-CHEMISTRY; PRIMATE; METABOLISM; HEMATOLOGY; BIOMARKERS; VARIABLES; RECEPTORS AB As a component of a long-term, longitudinal study of aging in this primate model, the objective of the current experiment was to assess age and diet effects on locomotor activity in a cross-sectional analysis. By attaching a motion detection device to the home cage, locomotor activity was monitored over a week in a group (N = 47) of female rhesus monkeys (Macaca mulatta) 6-26 Srs of age. About half these monkeys composed a control group fed a nutritionally fortified diet near ad libitum levels, whereas an experimental group had been fed the same diet at levels 30% less than comparable control levels for approximately 5 yrs prior to testing. Among control monkeys, a marked age-related decline in activity was noted when total activity was considered and also when diurnal and nocturnal periods of activity were analyzed separately. When comparing activity levels between control and experimental groups, only one significant diet effect was noted, which was in the youngest group of monkeys (6-8 yrs of age) during the diurnal period. Monkeys in the experimental group exhibited reduced activity compared to controls. Body weight was not consistently correlated to activity levels. In some older groups, heavier monkeys tended to show greater activity, but in younger groups the opposite pattern was observed. C1 NIA, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. ROW Sci, Gaithersburg, MD USA. Natl Inst Anim Hlth Ctr, Poolesville, MD USA. NIH, Off Res Serv, Vet Resources Program, Bethesda, MD 20892 USA. RP Ingram, DK (reprint author), NIA, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 40 TC 18 Z9 19 U1 0 U2 3 PU GERONTOLOGICAL SOCIETY AMER PI WASHINGTON PA 1275 K STREET NW SUITE 350, WASHINGTON, DC 20005-4006 USA SN 1079-5006 J9 J GERONTOL A-BIOL JI J. Gerontol. Ser. A-Biol. Sci. Med. Sci. PD AUG PY 2000 VL 55 IS 8 BP B373 EP B380 PG 8 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373WA UT WOS:000165312000002 PM 10952358 ER PT J AU Visser, M Harris, TB Fox, KM Hawkes, W Hebel, JR YuYahiro, J Michael, R Zimmerman, SI Magaziner, J AF Visser, M Harris, TB Fox, KM Hawkes, W Hebel, JR YuYahiro, J Michael, R Zimmerman, SI Magaziner, J TI Change in muscle mass and muscle strength after a hip fracture: Relationship to mobility recovery SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID X-RAY ABSORPTIOMETRY; SOFT-TISSUE COMPOSITION; BODY-COMPOSITION; FUNCTIONAL RECOVERY; GRIP STRENGTH; WOMEN; OLDER; MEN; PERFORMANCE; SARCOPENIA AB Background. Hip fracture in elderly persons has a serious impact on long-term physical function. This study determines the change in muscle strength and muscle mass after a hip fracture, and the associations between these changes and mobility recovery. Methods. Ninety community-dwelling women aged 65 years and older who had recently experienced a fracture of the proximal femur were included in the study. At 2 to 10 days after hospital admission, the women's grip strength, ankle dorsiflexion strength, and regional muscle mass (by dual-energy x-ray absorptiometry) were measured, and the prefracture level of independence for five mobility function items was assessed. All measurements were repented at 12 months. Results. At follow-up, only 17.8% of the women had returned to their prefracture level of mobility function for all Five items. Mobility function recovery was not related to change in skeletal muscle mass of the nonfractured leg or the arms. However, women who lost grip strength (mean loss of - 28.7%, SD = 16.9%), or who lost ankle strength of the nonfractured leg (mean loss of -21.5%. SD = 14.7%), had a worse mobility recovery compared with those who gained strength ((p =.04 and p =.09, respectively). In addition, chronic disease (p =.03), days hospitalized (p =.04), anti self-reported hip pain (p =.07) were independent predictors of decline in mobility function. Conclusions. The results suggest that loss of muscle strength, but not loss of muscle mass, is an independent predictor of poorer mobility recovery 12 months after a hip fracture. When confirmed by other studies, these findings may have implications fur rehabilitation strategies after a hip fracture. C1 Free Univ Amsterdam, Fac Med, Inst Res Extramural Med, NL-1081 BT Amsterdam, Netherlands. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. Euro RSCG, ManagedEDGE, New York, NY USA. Univ Maryland, Sch Med, Dept Epidemiol & Prevent Med, Baltimore, MD 21201 USA. RP Visser, M (reprint author), Free Univ Amsterdam, Fac Med, Inst Res Extramural Med, Boechorststr 7, NL-1081 BT Amsterdam, Netherlands. FU NIA NIH HHS [R37 AG09901] NR 34 TC 65 Z9 66 U1 1 U2 2 PU GERONTOLOGICAL SOCIETY AMER PI WASHINGTON PA 1275 K STREET NW SUITE 350, WASHINGTON, DC 20005-4006 USA SN 1079-5006 J9 J GERONTOL A-BIOL JI J. Gerontol. Ser. A-Biol. Sci. Med. Sci. PD AUG PY 2000 VL 55 IS 8 BP M434 EP M440 PG 7 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373WA UT WOS:000165312000009 PM 10952365 ER PT J AU Blagosklonny, MV AF Blagosklonny, MV TI Spotlight on apoptosis SO LEUKEMIA LA English DT Editorial Material C1 NCI, Med Branch, NIH, Bethesda, MD 20982 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bethesda, MD 20982 USA. NR 0 TC 2 Z9 3 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0887-6924 J9 LEUKEMIA JI Leukemia PD AUG PY 2000 VL 14 IS 8 BP 1500 EP 1501 DI 10.1038/sj.leu.2401863 PG 2 WC Oncology; Hematology SC Oncology; Hematology GA 341CB UT WOS:000088572500025 PM 10942249 ER PT J AU Blagosklonny, MV AF Blagosklonny, MV TI Cell death beyond apoptosis SO LEUKEMIA LA English DT Review DE apoptosis; leukemia; chemotherapy ID DEPENDENT KINASE INHIBITOR; DRUG-INDUCED APOPTOSIS; ACUTE MYELOID-LEUKEMIA; CAMPTOTHECIN-INDUCED APOPTOSIS; TERMINAL PROLIFERATION ARREST; CHRONIC LYMPHOCYTIC-LEUKEMIA; BREAST-CANCER CELLS; WILD-TYPE P53; C-H-RAS; DNA FRAGMENTATION AB Though the term apoptosis was originated in pathology and developmental biology as an alternative to necrosis, the tissue necrosis with inflammation is irrelevant to cell culture conditions where apoptosis is mostly studied. Furthermore, no one single morphological feature is either necessary or sufficient to define apoptosis. The emerging biochemical definition, a cell death with caspase activation, allows the distinction of alternative forms of cell death. Thus, inhibition of caspases delays but does not prevent cell death. Slow cell death without caspase activation may nevertheless be associated with DNA fragmentation. Oncogenic Ras, Raf, and mitogen-activated kinases inhibit apoptosis by affecting the cytochrome C/caspase-9 pathway but may arrest growth and cause slow cell death with delayed DNA fragmentation. Such 'slow' cell death without caspase activation is often caused by chemotherapeutic drugs. Whether a cell will undergo apoptosis or slow death depends not only on a chemotherapeutic agent but also on the readiness of cellular caspases. Therefore, one can distinguish apoptosisprone (eg leukemia) vs apoptosis-resistant cells. Cell susceptibilities to spontaneous, starvation-induced and drug-induced apoptosis are correlated and characterize an apoptosis-prone phenotype. Finally, distinction of slow cell death allows rephrasing of a question regarding the goal of cancer therapy: apoptosis vs slow cell death, or cancer cell-selectivity regardless of the mode of cell death. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bldg 10,R 12N226, Bethesda, MD 20892 USA. NR 123 TC 103 Z9 112 U1 6 U2 17 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0887-6924 J9 LEUKEMIA JI Leukemia PD AUG PY 2000 VL 14 IS 8 BP 1502 EP 1508 DI 10.1038/sj.leu.2401864 PG 7 WC Oncology; Hematology SC Oncology; Hematology GA 341CB UT WOS:000088572500026 PM 10942250 ER PT J AU Sheikh, MS Fornace, AJ AF Sheikh, MS Fornace, AJ TI Death and decoy receptors and p53-mediated apoptosis SO LEUKEMIA LA English DT Review DE TNF-R superfamily; death domain ID NF-KAPPA-B; DRUG-INDUCED APOPTOSIS; TNF RECEPTOR; TUMOR-CELLS; P53; ACTIVATION; DOMAIN; PROTEIN; SIGNAL; GENE AB Recently, several tumor necrosis factor receptor 1 (TNF-R1) and Fas-related death receptors have been discovered and include DR3, DR4, DR5 and DR6, These receptors contain an extracellular region containing varying numbers of cysteine-rich domains and an intracellular region that contains the death domain, The death receptors are activated in a ligand-dependent or independent manner and transduce apoptotic signals via their respective intracellular death domains. In addition to death receptors, several decoy molecules have also been identified and include DcR1/TRID, DcR2/TRUNDD, DcR3 and osteo-protegrin (OPG), The decoy molecules do not transduce apoptotic signals but rather compete with the death receptors for ligand binding and thereby inhibit ligand-induced apoptosis, Recent evidence suggests that p53 upregulates the expression of death receptors Fas and DR5, and thus, may mediate apoptosis in part via Fas and/or DR5, However, p53 also regulates the expression of TRAIL decoy receptors DcR1/TRID and DR2/TRUNDD. Although the significance of p53-dependent regulation of decoy receptors remains unclear, evidence suggests that DcR1/TRUNDD appears to inhibit 53-mediated apoptosis, It is, therefore, possible that p53 may blunt its DR5-dependent apoptotic effects by controlling the levels of decoy receptors. C1 SUNY Upstate Med Univ, Dept Pharmacol, Syracuse, NY 13210 USA. NCI, Gene Response Sect, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Sheikh, MS (reprint author), SUNY Upstate Med Univ, Dept Pharmacol, 750 E Adams St, Syracuse, NY 13210 USA. RI Fornace, Albert/A-7407-2008 OI Fornace, Albert/0000-0001-9695-085X NR 49 TC 122 Z9 133 U1 0 U2 9 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0887-6924 J9 LEUKEMIA JI Leukemia PD AUG PY 2000 VL 14 IS 8 BP 1509 EP 1513 DI 10.1038/sj.leu.2401865 PG 5 WC Oncology; Hematology SC Oncology; Hematology GA 341CB UT WOS:000088572500027 PM 10942251 ER PT J AU Welniak, LA Tian, ZG Sun, R Keller, JR Richards, S Ruscetti, FW Murphy, WJ AF Welniak, LA Tian, ZG Sun, R Keller, JR Richards, S Ruscetti, FW Murphy, WJ TI Effects of growth hormone and prolactin on hematopoiesis SO LEUKEMIA & LYMPHOMA LA English DT Review DE prolactin; growth hormone; IGF-I; myelosuppression; hematopoiesis; transplantation ID BONE-MARROW TRANSPLANTATION; FACTOR-I; PROGENITOR CELLS; TRANSGENIC MICE; ERYTHROPOIESIS INVITRO; PARTIALLY COUNTERACTS; MYELOID PROGENITORS; LYMPHOID-CELLS; EXPRESSION; SURVIVAL AB The use of the neuroendocrine hormones growth hormone (GH) and prolactin (PRL) in preclinical models, demonstrating promotion of hematopoietic recovery and immune function, offers promise for several clinical situations. These hormones do not appear to produce the same extent of immune/hematopoietic effects when compared to conventional hematopoietic and immune stimulating cytokines (i.e. G-CSF or interleukin-2). However, their pleiotropic effects and limited toxicity after systemic administration makes them attractive to test in myeloablative situations. More work needs to be performed to understand the mechanism(s) of GH and PRL action, particularly with regard to hematopoietic progenitor cell expansion and differentiation both in normal and pathologic situations. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC, IRSP, Ft Detrick, MD 21702 USA. RP Murphy, WJ (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC, IRSP, Bldg 567,Rm 210, Ft Detrick, MD 21702 USA. RI Tian, Zhigang/J-3512-2013 FU NCI NIH HHS [N01-CO-56000] NR 40 TC 18 Z9 19 U1 0 U2 0 PU HARWOOD ACAD PUBL GMBH PI READING PA C/O STBS LTD, PO BOX 90, READING RG1 8JL, BERKS, ENGLAND SN 1042-8194 J9 LEUKEMIA LYMPHOMA JI Leuk. Lymphoma PD AUG PY 2000 VL 38 IS 5-6 BP 435 EP 445 DI 10.3109/10428190009059263 PG 11 WC Oncology; Hematology SC Oncology; Hematology GA 343RN UT WOS:000088715500001 PM 10953965 ER PT J AU Qi, CF Hori, M Coleman, AE Torrey, TA Taddesse-Heath, L Ye, BH Chattopadhyay, SK Hartley, JW Morse, HC AF Qi, CF Hori, M Coleman, AE Torrey, TA Taddesse-Heath, L Ye, BH Chattopadhyay, SK Hartley, JW Morse, HC TI Genomic organisation and expression of BCL6 in murine B-cell lymphomas SO LEUKEMIA RESEARCH LA English DT Article DE BCL6; mouse B-cell lymphoma; translocation; spectral karyotyping ID NON-HODGKINS-LYMPHOMA; HEALTH-ORGANIZATION CLASSIFICATION; FINGER ENCODING GENE; CHROMOSOMAL TRANSLOCATIONS; IMMUNODEFICIENCY SYNDROME; 5'-NONCODING REGION; NEOPLASTIC DISEASES; PROTEIN EXPRESSION; MOUSE; MICE AB BCL6 encodes a transcription factor deregulated by chromosomal translocations in human diffuse large cell B lymphomas (DLCL). This study was designed to determine whether Bcl6 might also be involved in lymphomas of mice. BCL6 protein was expressed at high levels in 90% or more of DLCL but not in low grade B lymphomas. Southern hybridisation studies demonstrated altered organisation of Bcl6 in three primary DLCL and the WEHI 231 B-cell lymphoma cell line but not in low grade tumours. Chromosomal painting and fluorescence in situ hybridisation (FISH) analyses of the WEHI 231 metaphase spreads revealed a T(5;16) translocation with Bcl6 on Chromosome 16 at the translocation breakpoint. Deregulated expression of BCL6 is thus likely to contribute to the genesis of DLCL of mice as well as of humans. Published by Elsevier Science Ltd. C1 NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. Albert Einstein Coll Med, Dept Cell Biol, New York, NY USA. RP Morse, HC (reprint author), NIAID, Immunopathol Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Morse, Herbert/0000-0002-9331-3705 FU NIAID NIH HHS [N01-AI45203] NR 58 TC 15 Z9 15 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 2000 VL 24 IS 8 BP 719 EP 732 DI 10.1016/S0145-2126(00)00028-X PG 14 WC Oncology; Hematology SC Oncology; Hematology GA 339WE UT WOS:000088500200007 PM 10936424 ER PT J AU Hamilton, J Greiner, R Salem, N Kim, HY AF Hamilton, J Greiner, R Salem, N Kim, HY TI n-3 fatty acid deficiency decreases phosphatidylserine accumulation selectively in neuronal tissues SO LIPIDS LA English DT Article ID RAT-BRAIN; DOCOSAHEXAENOIC ACID; MEMBRANE; CELLS; DIET; PHOSPHOLIPIDS; MITOCHONDRIA; PROTEIN; PHOSPHATIDYLETHANOLAMINE; PERFORMANCE AB We have previously shown that the docosahexaenoate (22:6n-3) status in membrane phospholipids influences the biosynthesis and accumulation of phosphatidylserine (PS) in brain microsomes and C6 glioma cells. In the present study, we investigated whether the observed effect of membrane docosahexaenoic acid status on PS accumulation is universal or occurs specifically in neuronal tissues. We observed that rat brain cortex, brain mitochondria, and olfactory bulb, where 22.6n-3 is highly concentrated, contain significantly higher levels of PS in comparison to liver and adrenal, where 22:6n-3 is a rather minor component. Phospholipid molecular species analysis revealed that in brain cortex, mitochondria, and olfactory bulb 18:0,22:6n-3 was the most abundant species representing 45-65% of total PS. In nonneuronal tissues such as liver and adrenal, 18:0,20:4n-6 was the major PS species. Dietary depiction of n-3 fatty acids during prenatal and postnatal developmental periods decreased the brain 22:6n-3 content by more than 80%, with a concomitant increase in 22:5n-6 in all tissues. Under these conditions, an approximately 30-35% reduction in total PS in rat brain cortex, brain mitochondria, and olfactory bulb was observed, while PS levels in liver and adrenal were unchanged. The observed reduction of PS content in neuronal membranes appears to be due to a dramatic reduction of 18:0,22:6n-3-PS without complete replacement by 18:0,22:5n-6-PS. These results establish that variations in membrane 22:6n-3 fatty acid composition have a profound influence on PS accumulation in neuronal tissues where 22:6n-3 is abundant. These data have implications in neuronal signaling events where PS is believed to play an important role. C1 NIAAA, LMBB, Sect Mass Spectrometry, NIH, Rockville, MD 20852 USA. RP Kim, HY (reprint author), NIAAA, LMBB, Sect Mass Spectrometry, NIH, 12420 Parklawn Dr,Rm 158, Rockville, MD 20852 USA. NR 50 TC 68 Z9 73 U1 0 U2 3 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 2000 VL 35 IS 8 BP 863 EP 869 DI 10.1007/S11745-000-0595-x PG 7 WC Biochemistry & Molecular Biology; Nutrition & Dietetics SC Biochemistry & Molecular Biology; Nutrition & Dietetics GA 349CL UT WOS:000089026200006 ER PT J AU Murphy, EJ Rosenberger, TA Patrick, CB Rapoport, SI AF Murphy, EJ Rosenberger, TA Patrick, CB Rapoport, SI TI Intravenously injected [1-C-14]arachidonje acid targets phospholipids, and [1-C-14]palmitic acid targets neutral lipids in hearts of awake rats SO LIPIDS LA English DT Article ID CALCIUM-INDEPENDENT PHOSPHOLIPASE-A2; POLYUNSATURATED FATTY-ACIDS; ARACHIDONIC-ACID; ADULT-RAT; CARDIAC MYOCYTES; LONG-CHAIN; METABOLISM; MYOCARDIUM; CELLS; PLASMALOGEN AB The differential uptake and targeting of intravenously infused [1-C-14]palmitic ([1-C-14]16:0) and [1-C-14]arachidonic ([1-C-14]20:4n-6) acids into heart lipid pools were determined in awake adult male rats. The fatty acid tracers were infused (170 mu Ci/kg) through the femoral vein at a constant rate of 0.4 mL/min over 5 min. At 10 min postinfusion, the rats were killed using pentobarbital. The hearts were rapidly removed, washed free of exogenous blood, and frozen in dry ice. Arterial blood was withdrawn over the course of the experiment to determine plasma radiotracer levels. lipids were extracted from heart tissue using a two-phase system, and total radioactivity was measured in the nonvolatile aqueous and organic fractions. Both fatty acid tracers had similar plasma curves, but were differentially distributed into heart lipid compartments. The extent of [1-C-14]20:4n-6 esterification into heart phospholipids, primarily choline glycerophospholipids, was elevated 1.5-fold compared to [1-C-14]16:0, The unilateral incorporation coefficient, k*, which represents tissue radioactivity divided by the integrated plasma radioactivity for heart phospholipid, was sevenfold greater for [1-C-14]20:4n-6 than for [1-C-14]16:0. In contrast, [1-C-14]16:0 was esterified mainly into heart neutral lipids, primarily triacylglycerols (TC), and was also found in the nonvolatile aqueous compartment. Thus, in rat heart, [1-C-14]20:4n-6, was primarily targeted for esterification into phospholipids, while [1-C-14]16:0 was targeted for esterification into TG or metabolized into nonvolatile aqueous components. C1 NIA, Sect Brain Physiol & Metab, NIH, Bethesda, MD 20892 USA. RP Murphy, EJ (reprint author), NIA, Sect Brain Physiol & Metab, NIH, Bldg 10,Room 6C103, Bethesda, MD 20892 USA. NR 42 TC 17 Z9 17 U1 0 U2 0 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 2000 VL 35 IS 8 BP 891 EP 898 DI 10.1007/S11745-000-0598-7 PG 8 WC Biochemistry & Molecular Biology; Nutrition & Dietetics SC Biochemistry & Molecular Biology; Nutrition & Dietetics GA 349CL UT WOS:000089026200009 PM 10984112 ER PT J AU Silva, AC Kim, SG Garwood, M AF Silva, AC Kim, SG Garwood, M TI Imaging blood flow in brain tumors using arterial spin labeling SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE angiogenesis; cerebral blood flow; glioma rat brain; tumor necrosis ID RAT-BRAIN; PARTITION-COEFFICIENTS; PERFUSION; WATER; PERMEABILITY; SPECTROSCOPY; EXCHANGE; VESSELS; MODEL; MICE AB Measurements of tumor blood flow (TBF) are important for understanding tumor physiology and can be valuable in selecting and evaluating therapies. Brain tumors typically present reduced blood flows compared to normal brain tissue. This study shows that the arterial spin labeling (ASL) technique can be used to measure TBF non-invasively in a rat glioma model. Results show that TBF in the core (36.3 +/- 18.9 ml/100g/min, n = 4) and peripheral regions (85.3 +/- 26.9 ml/100g/min, n = 4) of the tumor are significantly reduced and show considerable heterogeneity compared to cerebral blood flow (CBF) of normal brain tissue (147.7 +/- 31.1 ml/100g/min, n = 4), while T-1 in the tumor (2.6 +/- 0.1 sec) is significantly elevated compared to normal tissue T-1 (2.0 +/- 0.0 sec), These results strongly support the feasibility of using the ASL technique to evaluate different cancer treatment strategies, to monitor the effects of agents designed to modulate TBF and oxygenation (e.g., carbogen gas), and to assess and guide the use of anti-angiogenic agents. Magn Reson Med 44:169-173, 2000, (C) 2000 Wiley-Liss, Inc. C1 Univ Minnesota, Sch Med, Dept Radiol, Ctr Magnet Resonance Res, Minneapolis, MN 55455 USA. RP Silva, AC (reprint author), NINDS, Lab Funct & Mol Imaging, NIH, 9000 Rockville Pike,Bldg 10,Room B1D69, Bethesda, MD 20892 USA. RI Silva, Afonso/A-7129-2009; Kim, Seong-Gi/O-8480-2014 OI Kim, Seong-Gi/0000-0003-1960-4464 FU NCI NIH HHS [CA64338]; NCRR NIH HHS [RR08079] NR 28 TC 71 Z9 75 U1 1 U2 7 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD AUG PY 2000 VL 44 IS 2 BP 169 EP 173 DI 10.1002/1522-2594(200008)44:2<169::AID-MRM1>3.0.CO;2-U PG 5 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 340QH UT WOS:000088545600001 PM 10918313 ER PT J AU von Bulow, GU Russell, H Copeland, NG Gilbert, DJ Jenkins, NA Bram, RJ AF von Bulow, GU Russell, H Copeland, NG Gilbert, DJ Jenkins, NA Bram, RJ TI Molecular cloning and functional characterization of murine Transmembrane Activator and CAML Interactor (TACI) with chromosomal localization in human and mouse SO MAMMALIAN GENOME LA English DT Article ID APOPTOSIS; COMPLEX; BINDING; PROTEIN AB The human Taci gene (Transmembrane Activator and CAML. Interactor) encodes a recently discovered member of the Tumor Necrosis Factor Receptor family. TACI is expressed in B-lymphocytes and may act to regulate humoral immunity. To identify functionally important regions of the protein, we have isolated and characterized the murine homolog of the human Taci cDNA. The proteins display 61.5% similarity and 54.6% identity. Mouse TACI is a type III transmembrane protein, as judged by the lack of a cleaved signal sequence and its N-terminal extracellular exposure. The intracellular domains of the mouse and human proteins share a single, defined region of high sequence conservation (19 of 23 residues identical). This constitutes a novel domain that may play a part in the initiation of signal transduction through TACI. In support of this notion, mouse TACI was found to activate NFAT, NFkB, and AP1 transcription factors in a transient transfection assay. The Taci gene was localized to human Chromosome (Chr) 17p11 by fluorescence in situ hybridization. The murine homolog was localized by intraspecific backcross analysis to the middle of Chr 11, a region that is syntenic to human Chr 17p. This work identifies concerned domains within TACI that may mediate the cellular distribution and signal transduction function of the protein and extend the details of homology between mouse Chr 11 and human 17p. C1 Mayo Clin & Mayo Fdn, Dept Pediat & Adolescent Med, Rochester, MN 55905 USA. Mayo Clin & Mayo Fdn, Dept Immunol, Rochester, MN 55905 USA. St Jude Childrens Res Hosp, Dept Genet, Memphis, TN 38105 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Mammalian Genet Lab, Frederick, MD 21702 USA. RP Bram, RJ (reprint author), Mayo Clin & Mayo Fdn, Dept Pediat & Adolescent Med, 200 1st St SW, Rochester, MN 55905 USA. FU NCI NIH HHS [1R01CA76274] NR 15 TC 38 Z9 38 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD AUG PY 2000 VL 11 IS 8 BP 628 EP 632 DI 10.1007/s003350010125 PG 5 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 338ZN UT WOS:000088453300004 PM 10920230 ER PT J AU Legler, J Potosky, AL Gilliland, FD Eley, JW Stanford, JL AF Legler, J Potosky, AL Gilliland, FD Eley, JW Stanford, JL TI Validation study of retrospective recall of disease-targeted function - Results from the prostate cancer outcomes study SO MEDICAL CARE LA English DT Article DE validation; retrospective recall; quality of life; prostate cancer ID QUALITY-OF-LIFE; RADICAL RETROPUBIC PROSTATECTOMY; BIAS; MEN AB OBJECTIVES. This was an ancillary methodological study within the Prostate Cancer Outcomes Study (PCOS) to assess the validity of 6-month retrospective recall of prediagnostic disease-targeted function among men diagnosed with prostate cancer. METHODS. A convenience sample of 133 prostate cancer cases were administered a baseline questionnaire shortly after diagnosis that asked about prediagnostic urinary, sexual, and bowel function. They were surveyed again concerning the same items 6 months later and asked to recall their prediagnostic function. Reports of prediagnostic function obtained at baseline and 6 months are compared, as are measures of change derived from these reports. Percent agreement and weighted kappas are calculated to measure the extent of agreement. RESULTS. Over 70% of the men reported prediagnostic functioning at the highest level on 12 of 17 survey items. For each of these items, recall at 6 months was identical to the baseline survey response for greater than or equal to 69% of the men. The values of the weighted kappas for changes computed with baseline reports (prospective) and changes computed with 6-month recall (retrospective) ranged from 0.396 to 0.919 for the 17 individual items. Intraclass correlations for the retrospective versus prospective changes in the multi-item function scores were 0.828 for urinary, 0.618 for bowel, and 0.692 for sexual function. CONCLUSIONS. At baseline, men recently diagnosed with prostate cancer report few disease-related problems before diagnosis, and a high percentage of men recall this accurately 6 months later. There is reasonably high agreement between baseline and 6-month estimates of prediagnostic function and between prospective and retrospective measures of change over 6 months. C1 NCI, Appl Res Branch, Bethesda, MD 20892 USA. USC, Sch Med, Dept Prevent Med, Los Angeles, CA USA. Emory Univ, Rollins Sch Publ Hlth, Georgia Ctr Canc Stat, Atlanta, GA 30322 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. RP Legler, J (reprint author), NCI, Appl Res Branch, EPN Suite 4103,MSC 7359,6130 Execut Blvd,MSC 7344, Bethesda, MD 20892 USA. FU NCI NIH HHS [N01-PC-67010, N01-PC-67009, N01-PC-67007] NR 25 TC 48 Z9 48 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 2000 VL 38 IS 8 BP 847 EP 857 DI 10.1097/00005650-200008000-00008 PG 11 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 337ZP UT WOS:000088393900008 PM 10929996 ER PT J AU De Lucca, AJ Bland, JM Vigo, CB Jacks, TJ Peter, J Walsh, TJ AF De Lucca, AJ Bland, JM Vigo, CB Jacks, TJ Peter, J Walsh, TJ TI D-Cecropin B: proteolytic resistance, lethality for pathogenic fungi and binding properties SO MEDICAL MYCOLOGY LA English DT Article DE cecropin B; D-enantiomer; fungicidal ID ANTIFUNGAL PROTEIN; ANTIMICROBIAL PEPTIDES; BIOLOGICAL PROPERTIES; SEQUENCE HOMOLOGY; INSECT IMMUNITY; CDNA CLONING; PURIFICATION; HEMOLYMPH; LARVAE AB L-Cecropin B (LCB) is a potent fungicidal peptide that is subject to proteolytic degradation by extracellular enzymes produced by Aspergillus flavus. We hypothesized that D-cecropin B (DCB), containing all D-amino acids, should resist proteolysis while retaining its fungicidal and target specificities. DCB was synthesized by solid phase methods using Fmoc chemistry. In vitro, at pH 6.0, DCB was lethal against the germinating conidia of A. flavus (LD90, 25 mu M) and A. fumigatus (LD98, 2.5 mu M) and for nongerminating and germinating conidia of Fusarium moniliforme (LD98, 1.25 mu M) and F. oxysporum (LD95, 2.5 mu M) at concentrations similar to those previously reported for LCB. It was lethal fdr Candida albicans with an LD98 at 12.5 mu M. DCB was not active for the nongerminating conidia of A. fumigatus or A. flavus. Papain, trypsin, pepsin A and Staphylococcus aureus V8 protease degraded LCB but not DCB, Binding assays and circular dichroism showed DCB and LCB bound to cholesterol, ergosterol, beta-1,3-glucan, mannan and chitin, Data show that DCB retains the potent fungicidal properties of the L-form while being resistant to proteolytic enzymes that degrade the latter peptide. This study demonstrates that D-enantiomerization of cecropin B yields a novel fungicidal peptide, which resists proteolytic degradation and is lethal for pathogenic fungi. C1 USDA ARS, So Reg Res Ctr, New Orleans, LA 70124 USA. NCI, Pediat Oncol Branch, Immunocompromised Hosp Sect, Bethesda, MD 20892 USA. RP De Lucca, AJ (reprint author), USDA ARS, So Reg Res Ctr, 1100 Robert E Lee Blvd, New Orleans, LA 70124 USA. NR 39 TC 15 Z9 15 U1 1 U2 3 PU B I O S SCIENTIFIC PUBLISHERS LTD PI OXFORD PA 9 NEWTEC PLACE, MAGDALEN RD, OXFORD OX4 1RE, ENGLAND SN 1369-3786 J9 MED MYCOL JI Med. Mycol. PD AUG PY 2000 VL 38 IS 4 BP 301 EP 308 DI 10.1080/714030954 PG 8 WC Infectious Diseases; Mycology; Veterinary Sciences SC Infectious Diseases; Mycology; Veterinary Sciences GA 352ZB UT WOS:000089248300003 PM 10975698 ER PT J AU Clarke, LP AF Clarke, LP TI NCI initiative: Development of novel imaging technologies SO MEDICAL PHYSICS LA English DT Editorial Material C1 NCI, Biomed Imaging Program, Div Canc Treatment & Diag, NIH, Bethesda, MD 20892 USA. RP Clarke, LP (reprint author), NCI, Biomed Imaging Program, Div Canc Treatment & Diag, NIH, 6130 Execut Blvd,EPN 800,MSC 7400, Bethesda, MD 20892 USA. NR 0 TC 3 Z9 3 U1 0 U2 0 PU AMER INST PHYSICS PI MELVILLE PA 2 HUNTINGTON QUADRANGLE, STE 1NO1, MELVILLE, NY 11747-4501 USA SN 0094-2405 J9 MED PHYS JI Med. Phys. PD AUG PY 2000 VL 27 IS 8 BP 1699 EP 1701 DI 10.1118/1.1287646 PG 3 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 347KM UT WOS:000088928300001 PM 10984213 ER PT J AU Lemmer, JT Hurlbut, DE Martel, GF Tracy, BL Ivey, FM Metter, EJ Fozard, JL Fleg, JL Hurley, BF AF Lemmer, JT Hurlbut, DE Martel, GF Tracy, BL Ivey, FM Metter, EJ Fozard, JL Fleg, JL Hurley, BF TI Age and gender responses to strength training and detraining SO MEDICINE AND SCIENCE IN SPORTS AND EXERCISE LA English DT Article DE resistance training; weight training; disuse; aging; men; women ID JOINT ANGLE SPECIFICITY; CROSS-SECTIONAL AREA; MUSCLE STRENGTH; BASAL METABOLISM; OLDER ADULTS; YOUNG-WOMEN; SHORT-TERM; MEN; EXERCISE; HYPERTROPHY AB Purpose: The purpose of this study was to examine the effects of age and gender on the strength response to strength training (ST and detraining. Methods: Eighteen young (20-30 yr) and 23 older (65-75 yr) men and women had their one-repetition maximum (1 RM) and isokinetic strength measured before and after 9 wk of unilateral knee extension ST (3 d.wk(-1)) and 31 wk of detraining. Results: The young subjects demonstrated a significantly greater (P < 0.05) increase in 1 RM strength (34 +/- 3%; 73 +/- 5 vs 97 +/- 6 kg; P < 0.01) than the older subjects (28 +/- 3%; 60 +/- 4 vs 76 +/- 5 kg, P < 0.01). There were no significant differences in strength gains between men and women in either age group with 9 wk of ST or in strength losses with 31 wk of detraining. Young men and women experienced an 8 +/- 2% decline in 1 RM strength after 31 wk of detraining (97 +/- 6 vs 89 +/- 6 kg, P < 0.05). This decline was significantly less than the 14 +/- 2% decline in the older men and women (76 +/- 5 vs 65 +/- 4 kg, P < 0.05). This strength loss occurred primarily between 12 and 31 wk of detraining with a 6 +/- 2% and 13 +/- 2% decrease in the young and older subjects, respectively, during this period. Discussion: These results demonstrate that changes in 1 RM strength in response to both ST and detraining are affected by age. However, ST-induced increases in muscular strength appear to be maintained equally well in young and older men and women during 12 wk of detraining and are maintained above baseline levels even after 31 wk of detraining in young men, young women, and older men. C1 Univ Maryland, Coll Hlth & Human Performance, Dept Kinesiol, College Pk, MD 20742 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. Univ Maryland Eastern Shore, Dept Phys Therapy, Princess Anne, MD 21853 USA. Morton Plant Mease Hlth Care, Florida Geriat Res Program, Clearwater, FL 33756 USA. RP Hurley, BF (reprint author), Univ Maryland, Coll Hlth & Human Performance, Dept Kinesiol, College Pk, MD 20742 USA. RI Fozard, James Leonard/B-3660-2009 FU NIA NIH HHS [1-AG-42148] NR 37 TC 121 Z9 132 U1 1 U2 12 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0195-9131 J9 MED SCI SPORT EXER JI Med. Sci. Sports Exerc. PD AUG PY 2000 VL 32 IS 8 BP 1505 EP 1512 DI 10.1097/00005768-200008000-00021 PG 8 WC Sport Sciences SC Sport Sciences GA 342LJ UT WOS:000088645800021 PM 10949019 ER PT J AU Tangerman, A Wilcken, B Levy, HL Boers, GHJ Mudd, SH AF Tangerman, A Wilcken, B Levy, HL Boers, GHJ Mudd, SH TI Methionine transamination in patients with homocystinuria due to cystathionine beta-synthase deficiency SO METABOLISM-CLINICAL AND EXPERIMENTAL LA English DT Article ID ISOLATED PERSISTENT HYPERMETHIONINEMIA; ADENOSYLTRANSFERASE DEFICIENCY; DOMINANT INHERITANCE; AMINO-ACIDS; METABOLISM; PATHWAY; RAT; HOMOCYSTEINE; PYRIDOXINE; MUTATION AB To assess the ability of patients with homocystinuria due to cystathionine beta-synthase (CBS) deficiency to perform the reactions of the methionine transamination pathway, the concentrations of the products of this pathway were measured in plasma and urine, The results clearly demonstrate that CBS-deficient patients develop elevations of these metabolites once a threshold near 350 mu mol/L for the concurrent plasma methionine concentration is exceeded. The absence of elevated methionine transamination products previously reported among 16 CBS-deficient B6-responsive patients may now be attributed to the fact that in those patients the plasma methionine concentrations were below this threshold. The observed elevations of transamination products were similar to those observed among patients with isolated hypermethioninemia. Plasma homocyst(e)ine did not exert a consistent effect on transamination metabolites, and betaine appeared to effect transamination chiefly by its tendency to elevate methionine. Even during betaine administration, the transamination pathway does not appear to be a quantitatively major route for the disposal of methionine. Copyright (C) 2000 by W.B. Saunders Company. C1 NIMH, DIRP, LMB, Bethesda, MD 20892 USA. Univ Nijmegen Hosp, Dept Gastrointestinal & Liver Dis, Nijmegen, Netherlands. Univ Nijmegen Hosp, Dept Gen Internal Med, Nijmegen, Netherlands. New Childrens Hosp, Westmead, NSW, Australia. Childrens Hosp, Boston, MA 02115 USA. Harvard Univ, Sch Med, Boston, MA USA. RP Mudd, SH (reprint author), NIMH, DIRP, LMB, Bldg 36,Room 1B-08,36 Convent Dr,MSC 4034, Bethesda, MD 20892 USA. NR 34 TC 31 Z9 31 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 2000 VL 49 IS 8 BP 1071 EP 1077 DI 10.1053/meta.2000.7709 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 349DU UT WOS:000089019100019 PM 10954028 ER PT J AU Maric, D Maric, I Barker, JL AF Maric, D Maric, I Barker, JL TI Dual video microscopic imaging of membrane potential and cytosolic calcium of immunoidentified embryonic rat cortical cells SO METHODS-A COMPANION TO METHODS IN ENZYMOLOGY LA English DT Article DE development; central nervous system; oxonol; fura-2; video microscopy ID CEREBELLAR GRANULE CELLS; GLUTAMATE RECEPTORS; K+ CHANNELS; SPINAL-CORD; LYMPHOCYTES; GABA; PROLIFERATION; EXCITABILITY; MECHANISMS; ACTIVATION AB Membrane potential (MP) and cytosolic Ca2+ (Ca-c(2+)) constitute important components involved in the physiological regulation of a myriad of cell functions in eukaryotic organisms. In particular, during development of the central nervous system, both properties are thought to be important in the regulation of cell cycle, cell migration, cell differentiation, cell-cell communication, and naturally occurring cell death. However, obtaining insight into the precise relationship between these two parameters of cell function is relatively limited either by technical difficulties inherent in using electrical recordings of membrane properties in conjunction with optical imaging of single cells or by employing optical imaging of either one or another property alone. Here, we describe in detail a novel strategy to record changes in both MP and Ca-c(2+) from many intact single cells in a noninvasive manner using digital video microscopy. This method involves double-loading the cells with voltage- and calcium-sensitive fluorescent indicator dyes, green oxonol, and fura-2, which can be sequentially excited with a mercury are lamp filtered at appropriate wavelengths and their resulting emissions can be captured with an intensified charged-coupled device camera at Is intervals. As an example of the utility of dual-recording strategy, we present data on a distinct functional expression of excitable membrane and cytoplasmic calcium properties in and differentiating embryonic rat cerebral cortical cells. C1 NINDS, Neurophysiol Lab, NIH, Bethesda, MD 20892 USA. RP Maric, D (reprint author), NINDS, Neurophysiol Lab, NIH, Bldg 36,Room 2C02,36 Convent Dr, Bethesda, MD 20892 USA. NR 29 TC 13 Z9 13 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1046-2023 J9 METHODS JI Methods PD AUG PY 2000 VL 21 IS 4 BP 335 EP 347 DI 10.1006/meth.2000.1023 PG 13 WC Biochemical Research Methods; Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 343WH UT WOS:000088725100004 PM 10964578 ER PT J AU Kindt, TJ Said, WA Bowers, FS Mahana, W Zhao, TM Simpson, RM AF Kindt, TJ Said, WA Bowers, FS Mahana, W Zhao, TM Simpson, RM TI Passage of human T-cell leukemia virus type-1 during progression to cutaneous T-cell lymphoma results in myelopathic disease in an HTLV-1 infection model SO MICROBES AND INFECTION LA English DT Article DE HTLV-I infections, pathogenicity; HTLV-I infections, transmission; HTLV-I infections, lymphoma; T-cell, cutaneous; nervous system, virology; disease model, animals; molecular sequence data ID CHRONIC INFLAMMATORY ARTHROPATHY; TROPICAL SPASTIC PARAPARESIS; PAPUA-NEW-GUINEA; I PROVIRAL DNA; MYCOSIS-FUNGOIDES; MOLECULAR CLONE; MONOCLONAL INTEGRATION; SOLOMON-ISLANDS; TRANSGENIC MICE; SEZARY-SYNDROME AB Studies comparing functional differences in human T-cell leukemia virus type 1 (HTLV-1) clones that mediate distinct outcomes in experimentally infected rabbits, resulted in a dermatopathic smoldering adult T-cell leukemia/lymphoma following chronic infection with HTLV-1 strain RH/K34. During the 3.5 years' follow-up, HTLV-1 skin disease progressed to cutaneous T-cell lymphoma. When infection was passed to several naive rabbits, progressive paraparesis due to myelopathic neurodegeneration, analogous to HTLV-associated myelopathy, resulted in one of 4 transfusion recipients. Similar proviral loads were detected in the two diseases, regardless of stage of progression or tissue compartment of infection. Complete proviral sequences obtained from the donor and affected recipient aligned identically with each other and with the inoculated virus clone. Existence of disparate pathogenic outcomes following infectious transmission further extends the analogy of using rabbits to model human infection and disease. Although the experimental outcomes shown are limited by numbers of animals affected, they mimic the infrequency of HTLV-1 disease and authenticate epidemiological evidence of virus sequence stability regardless of disease phenotype. The findings suggest that further investigation of a possible role for HTLV-1 in some forms of cutaneous T-cell lymphoma is warranted. (C) 2000 Editions scientifiques et medicales Elsevier SAS. C1 NIH Twinbrook Facil, Immunogenet Lab, Rockville, MD 20852 USA. RP Simpson, RM (reprint author), NIAID, 12441 Parklawn Dr, Rockville, MD 20852 USA. NR 50 TC 5 Z9 5 U1 0 U2 0 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 1286-4579 J9 MICROBES INFECT JI Microbes Infect. PD AUG PY 2000 VL 2 IS 10 BP 1139 EP 1146 DI 10.1016/S1286-4579(00)01268-5 PG 8 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 357FY UT WOS:000089488900002 PM 11008104 ER PT J AU Boulianne, RP Liu, Y Aebi, M Lu, BC Kues, U AF Boulianne, RP Liu, Y Aebi, M Lu, BC Kues, U TI Fruiting body development in Coprinus cinereus: regulated expression of two galectins secreted by a non-classical pathway SO MICROBIOLOGY-SGM LA English DT Article DE basidiomycete; Coprinus; fruiting body; galectin; secretion ID EXTRACELLULAR-MATRIX PROTEINS; FIBROBLAST-GROWTH-FACTOR; MATING-TYPE; MUSCLE LECTIN; CELLS; LOCALIZATION; ADHESION; LIGHT; GENE; DIFFERENTIATION AB Fruiting body formation in the basidiomycete Coprinus cinereus is a developmental process that occurs as a response of the mycelium to external stimuli. First, localized, highly branched hyphal structures (knots) are formed as a reaction to nutritional depletion. Hyphal-knot formation is repressed by light; however, light signals are essential for the development of the hyphal knot into an embryonic fruiting body (primordium) as well as karyogamy, meiosis and fruiting body maturation. The role of the different environmental signals in the initial phases of fruiting body development was analysed. It was observed that two fungal galectins, Cgl1 and Cgl2, are differentially regulated during fruiting body formation. cgl2 expression initiated in early stages of fruiting body development (hyphal knot formation) and was maintained until maturation of the fruiting body, whereas cgl1 was specifically expressed in primordia and mature fruiting bodies. Immunofluorescence and immunoelectron microscopy studies detected galectins within specific fruiting body tissues. They localized in the extracellular matrix and the cell wall but also in membrane-bound bodies in the cytoplasm. Heterologous expression of Cgl2 in Saccharomyces cerevisiae indicated that secretion of this protein occurred independently of the classical secretory pathway. C1 Swiss Fed Inst Technol, Inst Mikrobiol, CH-8092 Zurich, Switzerland. Univ Guelph, Dept Mol Biol & Genet, Guelph, ON N1G 2W0, Canada. RP NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. EM kues@micro.biol.ethz.ch RI Kues, Ursula/B-8280-2008; OI Kues, Ursula/0000-0001-9180-4079 NR 65 TC 59 Z9 64 U1 0 U2 5 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AG, BERKS, ENGLAND SN 1350-0872 J9 MICROBIOL-SGM JI Microbiology-(UK) PD AUG PY 2000 VL 146 BP 1841 EP 1853 PN 8 PG 13 WC Microbiology SC Microbiology GA 344NC UT WOS:000088764900009 PM 10931889 ER PT J AU Cabib, E AF Cabib, E TI On the zymogenic character of chitin synthase 3 SO MICROBIOLOGY-UK LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Cabib, E (reprint author), NIH, Bldg 8,Room 403, Bethesda, MD 20892 USA. NR 4 TC 0 Z9 1 U1 0 U2 0 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AE, BERKS, ENGLAND SN 1350-0872 J9 MICROBIOL-UK JI Microbiology-(UK) PD AUG PY 2000 VL 146 BP 1760 EP 1761 PN 8 PG 2 WC Microbiology SC Microbiology GA 344NC UT WOS:000088764900002 PM 10931882 ER PT J AU Arsura, M Mercurio, F Oliver, AL Thorgeirsson, SS Sonenshein, GE AF Arsura, M Mercurio, F Oliver, AL Thorgeirsson, SS Sonenshein, GE TI Role of the I kappa B kinase complex in oncogenic Ras- and Raf-mediated transformation of rat liver epithelial cells SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID TUMOR-NECROSIS-FACTOR; GROWTH-FACTOR-BETA; TRANSCRIPTION FACTOR; TGF-BETA; CONSTITUTIVE ACTIVATION; INDUCED APOPTOSIS; TRANSGENIC MICE; CANCER CELLS; OVAL CELLS; ALPHA AB NF-kappa B/Rel factors have been implicated in the regulation of liver cell death during development, after partial hepatectomy, and in hepatocytes in culture. Rat liver epithelial cells (RLEs) display many biochemical and ultrastructural characteristics of oval cells, which are multipotent cells that can differentiate into mature hepatocytes. While untransformed RLEs undergo growth arrest and apoptosis in response to transforming growth factor beta 1 (TGF-beta 1) treatment, oncogenic Ras- or Raf-transformed RLEs are insensitive to TGF-beta 1-mediated growth arrest. Here me have tested the hypothesis that Ras- or Raf-transformed RLEs have altered NF-kappa B regulation, leading to this resistance to TGF-beta 1. We show that classical NF-kappa B is aberrantly activated in Ras- or Raf-transformed RLEs, due to increased phosphorylation and degradation of I kappa B-alpha protein. Inhibition of NF-kappa B activity with a dominant negative form of I kappa B-alpha restored TGF-beta 1-mediated cell killing of transformed RLEs. IKK activity mediates this hyperphosphorylation of I kappa B-alpha protein. As judged by kinase assays and transfection of dominant negative IKK-1 and IKK-2 expression vectors, NF-kappa B activation by Ras appeared to be mediated by both IKK-1 and IKK-2, while Raf-induced NF-kappa B activation was mediated by IKK-2. NF-kappa B activation in the Ras-transformed cells was mediated by both the Raf and phosphatidylinositol 3-kinase pathways, while in the Raf-transformed cells, NF-kappa B induction was mediated by the mitogen-activated protein kinase cascade. Last, inhibition of either IKK-1 or IKK-2 reduced focus-forming activity in Ras-transformed RLEs. Overall, these studies elucidate a mechanism that contributes to the process of transformation of liver cells by oncogene Ras and Raf through the I kappa B kinase complex leading to constitutive activation of NF-kappa B. C1 Boston Univ, Sch Med, Dept Biochem, Boston, MA 02118 USA. Signal Pharmaceut Inc, San Diego, CA 92121 USA. NCI, Expt Carcinogenesis Lab, Div Basic Sci, Bethesda, MD 20892 USA. RP Arsura, M (reprint author), Boston Univ, Sch Med, Dept Biochem, 715 Albany St, Boston, MA 02118 USA. FU NCI NIH HHS [R01 CA036355, CA78616, CA36355, R01 CA078616] NR 81 TC 108 Z9 109 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 15 BP 5381 EP 5391 DI 10.1128/MCB.20.15.5381-5391.2000 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 334LX UT WOS:000088188900003 PM 10891479 ER PT J AU Muller, J Cacace, AM Lyons, WE McGill, CB Morrison, DK AF Muller, J Cacace, AM Lyons, WE McGill, CB Morrison, DK TI Identification of B-KSR1, a novel brain-specific isoform of KSR1 that functions in neuronal signaling SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID ACTIVATED PROTEIN-KINASE; ATP-BINDING SITE; MAP KINASE; GENE ENCODES; RAS EFFECTORS; C-ELEGANS; TRANSDUCTION; SUPPRESSOR; CELLS; DIFFERENTIATION AB Kinase suppressor of Ras (KSR) is an evolutionarily conserved component of Ras-dependent signaling pathways. Here, we report the identification of E-KSR1, a novel splice variant of murine KSR1 that is highly expressed in brain-derived tissues. B-KSR1 protein is detectable in mouse brain throughout embryogenesis, is most abundant in adult forebrain neurons, and is complexed with activated mitogen-activated protein kinase (MAPK) and MEK in brain tissues. Expression of B-KSR1 in PC12 cells resulted in accelerated nerve growth factor (NGF)-induced neuronal differentiation and detectable epidermal growth factor (EGF)-induced neurite outgrowth. Sustained MAPK activity was observed in cells stimulated with either NGF or EGF, and all effects on neurite outgrowth could be blocked by the MEK inhibitor PD98059. In B-KSR1-expressing cells, the MAPK-B-KSR1 interaction was inducible and correlated with MAPK activation, while the MEK-B-KSR1 interaction was constitutive. Further examination of the MEK-B-KSR1 interaction revealed that all genetically identified loss-of-function mutations in the catalytic domain severely diminished MEK binding. Moreover, B-KSR1 mutants defective in MER binding were unable to augment neurite outgrowth. Together, these findings demonstrate the functional importance of MEK binding and indicate that B-KSR1 may function to transduce Ras-dependent signals that are required for neuronal differentiation or that are involved in the normal functioning of the mature central nervous system. C1 NCI, Regulat Cell Growth Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Div Neuropathol, Baltimore, MD 21205 USA. RP Morrison, DK (reprint author), NCI, Regulat Cell Growth Lab, Frederick Canc Res & Dev Ctr, POB B, Frederick, MD 21702 USA. NR 34 TC 66 Z9 70 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 15 BP 5529 EP 5539 DI 10.1128/MCB.20.15.5529-5539.2000 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 334LX UT WOS:000088188900016 PM 10891492 ER PT J AU Liu, J Yu, SH Litman, D Chen, WP Weinstein, LS AF Liu, J Yu, SH Litman, D Chen, WP Weinstein, LS TI Identification of a methylation imprint mark within the mouse Gnas locus SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID GS-ALPHA GENE; DNA METHYLATION; CPG ISLAND; MESSENGER-RNA; IGF2R GENE; REGION; H19; TRANSCRIPTS; EXPRESSION; PROTEIN AB The imprinted mouse gene Gnas produces the G protein alpha-subunit Gs alpha and several other gene products by using alternative promoters and first exons. G(s)alpha is maternally expressed in some tissues and biallelically expressed in most other tissues, while the gene products NESP55 and XL alpha s are maternally and paternally expressed, respectively. We investigated the mechanisms of Gnas imprinting. The G(s)alpha promoter and first exon are not methylated on either allele. A further upstream region (approximately from positions -3400 to -939 relative to the G(s)alpha translational start site) is methylated only on the maternal allele in all adult somatic tissues and in early postimplantation development. Within this region lies a fourth promoter and first exon (exon 1A) that generates paternal-specific mRNAs of unknown function. Exon 1A and G(s)alpha mRNAs have similar expression patterns, making competition between their promoters unlikely. Differential methylation in this region is established during gametogenesis, being present in oocytes and absent in spermatozoa, and is maintained in preimplantation E3.5d blastocysts. Therefore, this region is a methylation imprint mark. In contrast, differential methylation of the NESP55 and XL alpha s promoter regions (Nesp and Gnasxl) is not established during gametogenesis. The methylation imprint mark that we identified may be important for the tissue-specific imprinting of G(s)alpha. C1 NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Weinstein, LS (reprint author), NIDDKD, Metab Dis Branch, NIH, Bldg 10,Rm 8C101, Bethesda, MD 20892 USA. RI Weinstein, Lee/I-5575-2015; OI Weinstein, Lee/0000-0002-1899-5152 NR 49 TC 136 Z9 143 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 16 BP 5808 EP 5817 DI 10.1128/MCB.20.16.5808-5817.2000 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 340GF UT WOS:000088524200003 PM 10913164 ER PT J AU Ho, HTB Chung, SK Law, JWS Ko, BCB Tam, SCF Brooks, HL Knepper, MA Chung, SSM AF Ho, HTB Chung, SK Law, JWS Ko, BCB Tam, SCF Brooks, HL Knepper, MA Chung, SSM TI Aldose reductase-deficient mice develop nephrogenic diabetes insipidus SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID COLLECTING DUCT; COMPLICATIONS; VASOPRESSIN; KIDNEY; PATHOGENESIS; AQUAPORIN-2; PREVENTION; GENE; RAT AB Aldose reductase (ALR2) is thought to be involved in the pathogenesis of various diseases associated with diabetes mellitus, such as cataract, retinopathy, neuropathy, and nephropathy, However, its physiological functions are not well understood. We developed mice deficient in this enzyme and found that they had no apparent developmental or reproductive abnormality except that they drank and urinated significantly more than their wild-type littermates, These ALR2-deficient mice exhibited a partially defective urine concentrating ability, having a phenotype resembling that of nephrogenic diabetes insipidus. C1 Univ Hong Kong, Inst Mol Biol, Hong Kong, Hong Kong, Peoples R China. Queen Mary Hosp, Div Clin Biochem, Hong Kong, Hong Kong, Peoples R China. NHLBI, Renal Mech Sect, NIH, Bethesda, MD 20892 USA. RP Chung, SK (reprint author), Univ Hong Kong, Inst Mol Biol, S Wing,8-F Kadoorie Biol Sci Bldg,Pokfulam Rd, Hong Kong, Hong Kong, Peoples R China. OI Ko, Ben Chi Bun/0000-0003-2027-5899 NR 38 TC 69 Z9 71 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 16 BP 5840 EP 5846 DI 10.1128/MCB.20.16.5840-5846.2000 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 340GF UT WOS:000088524200006 PM 10913167 ER PT J AU Al-Awar, O Radhakrishna, H Powell, NN Donaldson, JG AF Al-Awar, O Radhakrishna, H Powell, NN Donaldson, JG TI Separation of membrane trafficking and actin remodeling functions of ARF6 with an effector domain mutant SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID ADP-RIBOSYLATION FACTOR; RECEPTOR-MEDIATED ENDOCYTOSIS; PHOSPHOLIPASE-D; PLASMA-MEMBRANE; DOWNSTREAM EFFECTOR; NUCLEOTIDE EXCHANGE; MAMMALIAN-CELLS; TARGET PROTEINS; RAC REGULATION; CHOLERA-TOXIN AB The ADP-ribosylation factor 6 (ARF6) GTPase has a dual function in cells, regulating membrane traffic and organizing cortical actin. ARF6 activation is required for recycling of the endosomal membrane back to the plasma membrane (PM) and also for ruffling at the PM induced by Rac. Additionally, ARF6 at the PM induces the formation of actin-containing protrusions. To identify sequences in ARF6 that are necessary for these distinct functions, we examined the behavior of a chimeric protein of ARF1 and ARF6. The 1-6 chimera (with the amino half of ARF1 and the carboxyl half of ARF6) localized like ARF6 in HeLa cells and moved between the endosome and PM, but it did not form protrusions, an ARF6 effector function. Two residues in the amino-terminal half of ARF6, Q37 and S38, when substituted into the 1-6 chimera allowed protrusion formation, whereas removal of these residues from ARF6 resulted in an inability to form protrusions. interestingly, expression of 1-6 in cells selectively inhibited protrusions induced by wild-type ARF6 but had no effect on ARF6-regulated membrane movement or Rac-induced ruffling. Thus, we have uncoupled two functions of ARF6, one involved in membrane trafficking, which is necessary for Rac ruffling, and another involved in protrusion formation. C1 NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Donaldson, JG (reprint author), Ctr Dr,MSC 0301,Bldg 3,Room BI-22, Bethesda, MD 20892 USA. NR 48 TC 63 Z9 63 U1 2 U2 4 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 16 BP 5998 EP 6007 DI 10.1128/MCB.20.16.5998-6007.2000 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 340GF UT WOS:000088524200021 PM 10913182 ER PT J AU Franklin, DS Godfrey, VI O'Brien, DA Deng, CX Xiong, Y AF Franklin, DS Godfrey, VI O'Brien, DA Deng, CX Xiong, Y TI Functional collaboration between different cyclin-dependent kinase inhibitors suppresses tumor growth with distinct tissue specificity SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID MICE LACKING; INK4A LOCUS; P53; P27(KIP1); GENE; TUMORIGENESIS; HYPERPLASIA; DEFICIENT; P57(KIP2); RB AB The presence of two families of seven distinct mammalian cyclin-dependent kinase (CDK) inhibitor genes is thought to mediate the complexity of connecting a variety of cellular processes to the cell cycle control pathway. The distinct pattern of tissue expression of CDK inhibitor genes suggests that they may function as tumor suppressors with different tissue specificities. To test this hypothesis, we have characterized two strains of double mutant mice lacking either p18(INK4c) and p27(KIP1) or p18(INK4c) and p21(CIP1/WAF1). Loss of both p18 and p27 function resulted in the spontaneous development by 3 months of age of at least eight different types of hyperplastic tissues and/or tumors in the pituitary, adrenals, thyroid, parathyroid, testes, pancreas, duodenum, and stomach, Six of these hyperplastic tissues and tumors were in endocrine organs, and several types of tumors routinely developed within the same animal, a phenotype reminiscent of that seen in combined human multiple endocrine neoplasia syndromes. The p18-p21 double null mice, on the other hand, developed pituitary adenomas, multifocal gastric neuroendocrine hyperplasia, and lung bronchioalveolar tumors later in life. G(1) CDK2 and CDK4 kinase activities were increased in both normal and neoplastic tissues derived from mice lacking individual CDK inhibitors and were synergistically stimulated by the simultaneous loss of two CDK inhibitors. This indicates that an increase in G(1) CDK kinase activity is a critical step during but is not sufficient for tumor growth. Our results suggest that functional collaborations between distinct CDK inhibitor genes are tissue specific and confer yet another level of regulation in cell growth control and tumor suppression. C1 Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Pathol & Lab Med, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Anat & Cell Biol, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Pediat, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA. Univ N Carolina, Program Mol Biol & Biotechnol, Chapel Hill, NC 27599 USA. NIDDKD, Biochem & Metab Lab, NIH, Bethesda, MD 20892 USA. RP Xiong, Y (reprint author), Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA. RI deng, chuxia/N-6713-2016 FU NCI NIH HHS [P30 CA016086, CA16086, CA68377, R01 CA068377]; NICHD NIH HHS [HD26485, R01 HD026485] NR 43 TC 217 Z9 219 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD AUG PY 2000 VL 20 IS 16 BP 6147 EP 6158 DI 10.1128/MCB.20.16.6147-6158.2000 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 340GF UT WOS:000088524200035 PM 10913196 ER PT J AU Dong, JS Qiu, HF Garcia-Barrio, M Anderson, J Hinnebusch, AG AF Dong, JS Qiu, HF Garcia-Barrio, M Anderson, J Hinnebusch, AG TI Uncharged tRNA activates GCN2 by displacing the protein kinase moiety from a bipartite tRNA-Binding domain SO MOLECULAR CELL LA English DT Article ID TRANSFER-RNA-SYNTHETASES; EIF-2-ALPHA KINASE; SACCHAROMYCES-CEREVISIAE; DROSOPHILA-MELANOGASTER; CRYSTAL-STRUCTURE; INITIATION; PHOSPHORYLATION; IDENTIFICATION; RECOGNITION; EXPRESSION AB Protein kinase GCN2 regulates translation in amino acid-starved cells by phosphorylating eIF2. GCN2 contains a regulatory domain related to histidyl-tRNA synthetase (HisRS) postulated to bind multiple deacylated tRNAs as a general sensor of starvation. In accordance with this model, GCN2 bound several deacylated tRNAs with similar affinities, and aminoacylation of tRNA(Phe) weakened its interaction with GCN2. Unexpectedly, the C-terminal ribosome binding segment of GCN2 (C-term) was required in addition to the HisRS domain for strong tRNA binding. A combined HisRS+ C-term segment bound to the isolated protein kinase (PK) domain in vitro, and tRNA impeded this interaction. An activating mutation (GCN2(C)-E803V) that weakens PK-C-term association greatly enhanced tRNA binding by GCN2. These results provide strong evidence that tRNA stimulates the GCN2 kinase moiety by preventing an inhibitory interaction with the bipartite tRNA binding domain. C1 NICHHD, Lab Eukaryot Gene Regulat, Bethesda, MD 20892 USA. RP Hinnebusch, AG (reprint author), NICHHD, Lab Eukaryot Gene Regulat, Bethesda, MD 20892 USA. NR 31 TC 157 Z9 165 U1 2 U2 15 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell. PD AUG PY 2000 VL 6 IS 2 BP 269 EP 279 DI 10.1016/S1097-2765(00)00028-9 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 351NY UT WOS:000089166100006 PM 10983975 ER PT J AU Intine, RVA Sakulich, AL Koduru, SB Huang, Y Pierstorff, E Goodier, JL Phan, L Maraia, RJ AF Intine, RVA Sakulich, AL Koduru, SB Huang, Y Pierstorff, E Goodier, JL Phan, L Maraia, RJ TI Control of transfer RNA maturation by phosphorylation of the human La antigen on Serine 366 SO MOLECULAR CELL LA English DT Article ID POLYMERASE-III TRANSCRIPTS; TERMINATION FACTOR LA; SACCHAROMYCES-CEREVISIAE; SCHIZOSACCHAROMYCES-POMBE; BINDING PROTEIN; XENOPUS-LAEVIS; AUTOANTIGEN LA; NUCLEAR IMPORT; S-POMBE; YEAST AB Conversion of a nascent precursor tRNA to a mature functional species is a multipartite process that involves the sequential actions of several processing and modifying enzymes. La is the first protein to interact with pre-tRNAs in eukaryotes. An opal suppressor tRNA served as a functional probe to examine the activities of yeast and human (h)La proteins in this process in fission yeast. An RNA recognition motif and Walker motif in the metazoan-specific C-terminal domain (CTD) of hLa maintain pre-tRNA in an unprocessed state by blocking the 5'-processing site, impeding an early step in the pathway. Faithful phosphorylation of hLa on serine 366 reverses this block and promotes tRNA maturation. The results suggest that regulation of tRNA maturation at the level of RNase P cleavage may occur via phosphorylation of serine 366 of hLa. C1 NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Maraia, RJ (reprint author), NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. NR 42 TC 64 Z9 70 U1 2 U2 2 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell. PD AUG PY 2000 VL 6 IS 2 BP 339 EP 348 DI 10.1016/S1097-2765(00)00034-4 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 351NY UT WOS:000089166100012 PM 10983981 ER PT J AU McKay, LI Cidlowski, JA AF McKay, LI Cidlowski, JA TI CBP (CREB binding protein) integrates NF-kappa B (nuclear factor-kappa B) and glucocorticoid receptor physical interactions and antagonism SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID UBIQUITIN-PROTEASOME PATHWAY; TRANSCRIPTIONAL ACTIVATION; ANTIINFLAMMATORY ACTION; FUNCTIONAL ANTAGONISM; DOWN-REGULATION; CROSS-TALK; ALPHA; MECHANISMS; INHIBITOR; COACTIVATORS AB Nuclear factor-kappa B (NF-kappa B) and the glucocorticoid receptor (GR) are transcription factors with opposing actions in the modulation of immune/inflammatory responses. NF-kappa B induces the expression of proinflammatory genes, while GR suppresses immune function in part by suppressing expression of the same genes. Previously, we demonstrated that physiological antagonism between NF-kappa B and GR is due to a mutual transcriptional antagonism that requires the p65 subunit of NF-kappa B and multiple domains of GR (1). To elucidate the mechanism(s) of NF-kappa B p65 and GR transcriptional antagonism, we analyzed the interactions of wild-type p65 and p65 RHD (rel homology domain, a dominant negative mutant of p65 which lacks a transactivation domain) with GR. We show that p65RHD blocks p65-mediated transactivation, yet does not block the repression of GR transactivation by p65, indicating that transcriptional activity by p65 is not required to repress GR function. Both p65 and p65 RHD physically interact with GR, but only intact p65 represses OR-mediated signaling, implicating the p65 transactivation domain in the transcriptional repression of GR. To further characterize p65-GR interactions, we examined the role of the transcriptional co-integrator CREB binding protein (CBP) in their mutual antagonism. OR-mediated repression of p65 transactivation and p65-mediated repression of GR transactivation, as well as the physical interaction between NF-kappa B and GR, are enhanced by CBP. GR bound to the antagonist RU 486, although transcriptionally inactive, retains the ability to repress p65 transactivation. However, CBP does not physically interact with antagonist-bound GR and does not enhance its repressive effect on p65. These data suggest that CBP functions as an integrator of p65/GR physical interaction, rather than as a limiting cofactor for which p65 and GR compete. C1 NIEHS, Mol Endocrinol Grp, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Cidlowski, JA (reprint author), NIEHS, Mol Endocrinol Grp, Lab Signal Transduct, NIH, POB 12233,MD E2-02, Res Triangle Pk, NC 27709 USA. EM Cidlowski@NIEHS.NIH.GOV NR 42 TC 110 Z9 117 U1 0 U2 3 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD AUG PY 2000 VL 14 IS 8 BP 1222 EP 1234 DI 10.1210/me.14.8.1222 PG 13 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 339VK UT WOS:000088498400009 PM 10935546 ER PT J AU Zhang, ZJ Mandal, AK Mital, A Popescu, N Zimonjic, D Moser, A Moser, H Mukherjee, AB AF Zhang, ZJ Mandal, AK Mital, A Popescu, N Zimonjic, D Moser, A Moser, H Mukherjee, AB TI Human acid ceramidase gene: Novel mutations in Farber disease SO MOLECULAR GENETICS AND METABOLISM LA English DT Article DE acid ceramidase (AC); Farber disease (FD); lipogranulomatosis; gene mutations; ceramide ID PROTEIN; CLONING; LOCALIZATION; EXPRESSION; TUMORS; LUNG AB Farber disease is an autosomal recessive disorder caused by lysosomal acid ceramidase (AC) deficiency, It commonly manifests during the first few months after birth with a unique triad of painful and progressive deformed joints, subcutaneous nodules, and progressive hoarseness, In order to understand the molecular mechanism(s) of pathogenesis of Farber disease, we isolated and characterized a full-length human AC gene, mapped its chromosomal location, determined the tissue-specific expression, and analyzed mutations in Farber disease patients. We also studied the AC-mRNA expression in gastrointestinal tumors and adjoining normal tissues. In addition, we determined the pattern of tissue-specific AC-mRNA expression in the adult mouse and during fetal development. Our results show that human AC gene consists of 14 exons and 13 introns spanning approximately 26.5 kb of genomic DNA. It is mapped to human chromosome 8p22-21.2, a region often disrupted in several cancers. The AC-mRNA is expressed in the mouse fetus from the seventh day of gestation. Interestingly, while the AC-mRNA is expressed in all segments of the normal gastrointestinal tract, none of the gastrointestinal tumor tissues had any AC-mRNA expression. We also uncovered four novel mutations in Farber disease patients that were not previously reported. Taken together, our results not only attest to the physiological importance of AC but also uncover several new mutations in Farber disease that may advance our knowledge towards establishing a genotype-phenotype correlation in this disease. (C) 2000 Academic Press. C1 NICHD, Sect Dev Genet, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. NCI, Lab Mol Cytogenet, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Kennedy Krieger Inst, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Neurol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Pediat, Baltimore, MD 21205 USA. RP Mukherjee, AB (reprint author), Bldg 10,Room 9S241, Bethesda, MD 20892 USA. NR 28 TC 19 Z9 20 U1 1 U2 1 PU ACADEMIC PRESS INC 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 2000 VL 70 IS 4 BP 301 EP 309 DI 10.1006/mgme.2000.3029 PG 9 WC Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental SC Endocrinology & Metabolism; Genetics & Heredity; Research & Experimental Medicine GA 394JJ UT WOS:000166520000007 PM 10993717 ER PT J AU Mimura, Y Church, S Ghirlando, R Ashton, PR Dong, S Goodall, M Lund, J Jefferis, R AF Mimura, Y Church, S Ghirlando, R Ashton, PR Dong, S Goodall, M Lund, J Jefferis, R TI The influence of glycosylation on the thermal stability and effector function expression of human IgG1-Fc: properties of a series of truncated glycoforms SO MOLECULAR IMMUNOLOGY LA English DT Article DE differential scanning microcalorimetry; Fc gamma receptor; complement; electrospray ionisation mass spectrometry ID FC-GAMMA-RECEPTOR; HUMAN-IGG; CRYSTAL-STRUCTURE; OLIGOSACCHARIDE CHAINS; CARBOHYDRATE STRUCTURE; SCANNING CALORIMETRY; IMMUNOGLOBULIN-G; FRAGMENT; ANTIBODY; AFFINITY AB Antibodies are multifunctional molecules that following the formation of antibody antigen complexes, may activate mechanisms to effect the clearance and destruction of the antigen (pathogen). The IgG molecule is comprised of three globular protein moieties (2Fab + Fc) linked through a flexible hinge region. While the Fabs bind antigens, the Fc triggers effector mechanisms through interactions with specific ligands, e.g, cellular receptors (Fc gammaR), and the C1 component of complement. Glycosylation of IgG-Fc has been shown to be essential for efficient activation of Fc gammaR and C1. We report the generation of a series of truncated glycoforms of IgG-Fc, and the analysis of the contribution of the residual oligosaccharide to IgG-Fc function and thermal stability. Differential scanning microcalorimetry has been used to compare the stabilities of the homogeneous glycoforms of IgG1-Fc. The results show that all truncated oligosaccharides confer a degree of functional activity, and thermodynamic stability to the IgG1-Fc, in comparison with deglycosylated IgG1-Fc. The same truncated glycoforms of an intact IgG1 anti-MHC Class II antibody are shown to exhibit differential functional activity for Fc gamma RI and C1 ligands, relative to deglycosylated IgG1. The minimal glycoform investigated had a trisaccharide attached to each heavy chain and can be expected to influence protein structure primarily in the proximity of the N-terminal region of the C(H)2 domain, implicated as a binding site for multiple effector ligands. These data provide a thermodynamic rationale for the modulation of antibody effector functions by different glycoforms. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Univ Birmingham, Sch Med, Div Immun & Infect, Birmingham B15 2TT, W Midlands, England. NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Birmingham, Sch Chem, Birmingham B15 2TT, W Midlands, England. RP Jefferis, R (reprint author), Univ Birmingham, Sch Med, Div Immun & Infect, Birmingham B15 2TT, W Midlands, England. RI Ghirlando, Rodolfo/A-8880-2009 NR 36 TC 170 Z9 174 U1 2 U2 13 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0161-5890 J9 MOL IMMUNOL JI Mol. Immunol. PD AUG-SEP PY 2000 VL 37 IS 12-13 BP 697 EP 706 DI 10.1016/S0161-5890(00)00105-X PG 10 WC Biochemistry & Molecular Biology; Immunology SC Biochemistry & Molecular Biology; Immunology GA 413ML UT WOS:000167615900002 PM 11275255 ER PT J AU Diaw, L Siwarski, D DuBois, W Jones, G Huppi, K AF Diaw, L Siwarski, D DuBois, W Jones, G Huppi, K TI Double producers of kappa and lambda define a subset of B cells in mouse plasmacytomas SO MOLECULAR IMMUNOLOGY LA English DT Article DE plasmacytoma; allelic exclusion; single cell micromanipulation; dual light chain ID LIGHT-CHAIN GENES; ALLELIC EXCLUSION; MESSENGER-RNA; SINGLE CELLS; EXPRESSION; REARRANGEMENT; RECOMBINATION; FREQUENCY; HYBRIDOMA; LYMPHOMA AB Rearrangement of the light chain locus is believed to be an ordered process in which Ig lambda rearrangements only occur if Ig kappa rearrangements are found to be non-productive or self-reactive. Secondary rearrangements of the B-cell receptor (BCR) have shown, however, that rescue of abortive Ig kappa rearrangements or autoreactive B cells can be achieved through receptor editing using upstream V-regions as the template sequences. Since secondary rearrangement can occur in the periphery, possibly in a subset of B cells maintaining constitutive Rag activity, it is conceivable that two light chains (kappa:kappa or kappa:lambda) could be expressed in these cells, apparently in violation of allelic exclusion. Previously, we have reported that silicone-induced plasmacytomas (SIPCs) exhibit dual expression and ongoing rearrangements of Ig kappa and Ig lambda. In this paper, we show by ELISA that both Ig kappa and Ig lambda are found at the protein level, but are secreted in different amounts. Furthermore, we demonstrate by micro-manipulation and RT-PCR amplification that Ig kappa and Ig lambda are simultaneously expressed in a single SIPC cell. We propose that these dual-expressing cells, found intermittently in cases of plasmacytomas (PCs), may have originally been immature B cells when transformed but now are maintained as a long-lived mature B cell found infrequently in the tumor population. (C) 2001 Published by Elsevier Science Ltd. C1 NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. RP Huppi, K (reprint author), NCI, Genet Lab, NIH, Bldg 37,Rm 2B-21, Bethesda, MD 20892 USA. NR 48 TC 13 Z9 14 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0161-5890 J9 MOL IMMUNOL JI Mol. Immunol. PD AUG-SEP PY 2000 VL 37 IS 12-13 BP 775 EP 781 DI 10.1016/S0161-5890(00)00100-0 PG 7 WC Biochemistry & Molecular Biology; Immunology SC Biochemistry & Molecular Biology; Immunology GA 413ML UT WOS:000167615900009 PM 11275262 ER PT J AU Chattoraj, DK AF Chattoraj, DK TI Control of plasmid DNA replication by iterons: no longer paradoxical SO MOLECULAR MICROBIOLOGY LA English DT Review ID MINI-F PLASMID; INITIATOR PROTEIN REPE; ESCHERICHIA-COLI; COPY-NUMBER; RK2 REPLICATION; CHROMOSOMAL ORIGIN; NEGATIVE CONTROL; BINDING-SITES; PSC101; P1 AB Replication origins of a family of bacterial plasmids have multiple sites, called iterons, for binding a plasmid-specific replication initiator protein. The iteron-initiator interactions are essential for plasmid replication as well as for inhibition of plasmid overreplication. The inhibition increases with plasmid copy number and eventually shuts plasmid replication off completely. The mechanism of inhibition appears to be handcuffing, the coupling of origins via iteron-bound initiators that block origin function. The probability of a trans-reaction such as handcuffing is expected to increase with plasmid copy number and diminish with increases in cell volume, explaining how the copy number can be maintained in a growing cell. Control is also exerted at the level of initiator synthesis and activation by chaperones. We propose that increases in active initiators promote initiation by overcoming handcuffing, but handcuffing dominates when the copy number reaches a threshold. Handcuffing should be ultrasensitive to copy number, as the negative control by iterons can be stringent (switch-like). C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Chattoraj, DK (reprint author), NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. NR 72 TC 124 Z9 126 U1 5 U2 16 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD AUG PY 2000 VL 37 IS 3 BP 467 EP 476 DI 10.1046/j.1365-2958.2000.01986.x PG 10 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 345FV UT WOS:000088806700002 PM 10931340 ER PT J AU Shaw, EI Dooley, CA Fischer, ER Scidmore, MA Fields, KA Hackstadt, T AF Shaw, EI Dooley, CA Fischer, ER Scidmore, MA Fields, KA Hackstadt, T TI Three temporal classes of gene expression during the Chlamydia trachomatis developmental cycle SO MOLECULAR MICROBIOLOGY LA English DT Article ID OUTER-MEMBRANE PROTEIN; ABC FAMILY TRANSPORTER; ESCHERICHIA-COLI; INCLUSION MEMBRANE; NUCLEOTIDE-SEQUENCE; MURAMIC ACID; SUCCINATE-DEHYDROGENASE; COMPETITIVE PCR; INFECTED-CELLS; PSITTACI AB The obligate intracellular bacterium Chlamydia trachomatis has a unique developmental cycle that involves functionally and morphologically distinct cell types adapted for extracellular survival and intracellular multiplication. Infection is initiated by an environmentally resistant cell type called an elementary body (EB). Over the first several hours of infection, EBs differentiate into a larger replicative form, termed the reticulate body (RB). Late in the infectious process, RBs asynchronously begin to differentiate back to EBs, which accumulate within the lumen of the inclusion until released from the host cell for subsequent rounds of infection. In an effort to characterize temporal gene expression in relation to the chlamydial developmental cycle, we have used quantitative-competitive polymerase chain reaction (QC-PCR) and reverse transcription (RT)-PCR techniques. These analyses demonstrate that C. trachomatis double their DNA content every 2-3 h, with synthesis beginning between 2 and 4 h after infection. We determined the onset of transcription of specific temporal classes of developmentally expressed genes. RT-PCR analysis was performed on several genes encoding key enzymes or components of essential biochemical pathways and functions. This comparison encompassed approximately 8% of open reading frames on the C. trachomatis genome. In analysis of total RNA samples harvested at 2, 6, 12 and 20 h after infection, using conditions under which a single chlamydial transcript per infected cell is detected, three major temporal classes of gene expression were resolved. Initiation of transcription appears to occur in three temporal classes which we have operationally defined as: early, which are detected by 2 h after infection during the germination of EBs to RBs; mid-cycle, which appear between 6 and 12 h after infection and represent transcripts expressed during the growth and multiplication of RBs; or late, which appear between 12 and 20 h after infection and represent those genes transcribed during the terminal differentiation of RBs to EBs. Collectively, the data suggest that chlamydial early gene functions are weighted toward initiation of macromolecular synthesis and the establishment of their intracellular niche by modification of the inclusion membrane. Surprisingly, representative enzymes of intermediary metabolism and structural proteins do not appear to be transcribed until 10-12 h after infection; coinciding with the onset of observed binary fission of RBs. Late gene functions appear to be predominately those associated with the terminal differentiation of RBs back to EBs. C1 NIAID, Rocky Mt Labs, Host Parasite Interact Sect, Intracellular Parasites Lab, Hamilton, MT 59840 USA. NIAID, Rocky Mt Labs, Microscopy Branch, Hamilton, MT 59840 USA. RP Hackstadt, T (reprint author), NIAID, Rocky Mt Labs, Host Parasite Interact Sect, Intracellular Parasites Lab, Hamilton, MT 59840 USA. NR 69 TC 139 Z9 143 U1 1 U2 10 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD AUG PY 2000 VL 37 IS 4 BP 913 EP 925 DI 10.1046/j.1365-2958.2000.02057.x PG 13 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 349PD UT WOS:000089054100020 PM 10972811 ER PT J AU Martin, RG Gillette, WK Rosner, JL AF Martin, RG Gillette, WK Rosner, JL TI The ykgA gene of Escherichia coli SO MOLECULAR MICROBIOLOGY LA English DT Letter ID TRANSCRIPTIONAL ACTIVATOR; SEQUENCE C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Martin, RG (reprint author), NIDDKD, Mol Biol Lab, NIH, Bldg 5,Room 333, Bethesda, MD 20892 USA. NR 6 TC 6 Z9 6 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD AUG PY 2000 VL 37 IS 4 BP 978 EP 979 DI 10.1046/j.1365-2958.2000.02043.x PG 2 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 349PD UT WOS:000089054100025 PM 10972816 ER PT J AU Tsao, CC Foley, J Coulter, SJ Maronpot, R Zeldin, DC Goldstein, JA AF Tsao, CC Foley, J Coulter, SJ Maronpot, R Zeldin, DC Goldstein, JA TI CYP2C40, a unique arachidonic acid 16-hydroxylase, is the major CYP2C in murine intestinal tract SO MOLECULAR PHARMACOLOGY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; EPOXYEICOSATRIENOIC ACIDS; EXTRAHEPATIC TISSUES; IMMUNOHISTOCHEMICAL LOCALIZATION; CYTOCHROME-P450; RAT; METABOLITES; EXPRESSION; CYTOCHROMES-P-450; IDENTIFICATION AB We recently identified five different murine CYP2C cDNAs from a murine cDNA library. When expressed in a bacterial cDNA expression system, all five recombinant proteins metabolized arachidonic acid but produced distinctly different profiles. In addition, some CYP2C mRNAs were found in extrahepatic tissues, as well as in liver. Immunoblots with an antibody raised against recombinant CYP2C38, which recognizes all five murine CYP2Cs, demonstrated that among extrahepatic tissues, colon and cecum contained the highest amount of CYP2Cs. The highest concentration of CYP2Cs occurred in cecum and colon (cecum greater than or equal to proximal colon much greater than distal colon), with lower levels in duodenum, jejunum, and ileum. Immunohistochemical studies revealed that CYP2Cs were localized principally in epithelial cells and autonomic ganglia in gut and colon. Polymerase chain reaction amplification of reverse-transcribed mRNA using murine CYP2C-specific primers followed by cloning and sequencing identified CYP2C40 as the major CYP2C isoform expressed in murine intestinal tract. Recombinant CYP2C40 metabolized arachidonic acid in a regio- and stereospecific manner to 16(R)-HETE (hydroxyeicosatetraenoic acid) as the major product. To our knowledge, CYP2C40 is the first enzyme known to produce primarily 16-HETE. We conclude that CYP2C40 is one of the major cytochrome P450 proteins in the mouse intestinal tract. In the light of vasoactive and anti-neutrophilic effects of 16-HETE, we hypothesize that CYP2C40 may play an important role in endogenous biological functions in intestine. C1 NIEHS, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. NIEHS, Pulm Pathobiol Lab, Res Triangle Pk, NC 27709 USA. RP NIEHS, Lab Pharmacol & Chem, 111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. EM goldste1@niehs.nih.gov RI Goldstein, Joyce/A-6681-2012; OI Coulter, Sherry/0000-0002-2732-3470 NR 44 TC 22 Z9 22 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0026-895X EI 1521-0111 J9 MOL PHARMACOL JI Mol. Pharmacol. PD AUG PY 2000 VL 58 IS 2 BP 279 EP 287 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 337JT UT WOS:000088354900005 PM 10908295 ER PT J AU Dunbar, C AF Dunbar, C TI Hematopoietic stem cells and gene transfer SO MOLECULAR THERAPY LA English DT Article C1 NHLBI, NIH, Bethesda, MD 20892 USA. RP Dunbar, C (reprint author), NHLBI, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC 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 2000 VL 2 IS 2 BP 98 EP 98 PG 1 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 367EJ UT WOS:000090048500005 ER PT J AU Bodine, D AF Bodine, D TI Globin gene therapy: One (Seemingly) small vector change, one giant leap in optimism SO MOLECULAR THERAPY LA English DT Editorial Material ID HEMATOPOIETIC STEM-CELLS; LOCUS-CONTROL REGION; RETROVIRAL VECTORS; EXPRESSION; MICE; DERIVATIVES C1 NHGRI, NIH, Bethesda, MD 20892 USA. RP Bodine, D (reprint author), NHGRI, NIH, 49 Convent Dr,MSC 4442, Bethesda, MD 20892 USA. NR 15 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC 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 2000 VL 2 IS 2 BP 101 EP 102 DI 10.1006/mthe.2000.0113 PG 2 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 367EJ UT WOS:000090048500007 PM 10947935 ER PT J AU Tranquill, LR Cao, LG Ling, NC Kalbacher, H Martin, RM Whitaker, JN AF Tranquill, LR Cao, LG Ling, NC Kalbacher, H Martin, RM Whitaker, JN TI Enhanced T cell responsiveness to citrulline-containing myelin basic protein in multiple sclerosis patients SO MULTIPLE SCLEROSIS JOURNAL LA English DT Article DE multiple sclerosis; MBP; citrulline; T cells; human; isomer ID HEALTHY-INDIVIDUALS; HLA RESTRICTION; PEPTIDE; RECOGNITION; GENE; MBP; ENCEPHALOMYELITIS; AUTOIMMUNITY; RESPONSES; CONTEXT AB Myelin basic protein (MBP), a candidate autoantigen in multiple sclerosis (MS), exists in different isoforms and charge isomers generated by differential splicing of exons and by a combination of posttranslational modifications respectively These various isoforms and charge isomers of MBP vary in abundance and most likely serve different functions during myelinogenesis and remyelination. The least cationic among the charge isomers of MBP is citrullinated and is referred to as MBP-C8. MBP-C8 is relatively increased in the population of MBP isomers in more developmentally immature myelin and in MS brain tissue. In a previous study, we found that MBP-C8-reactive T cells could be detected in CD4+ T cell lines (TCL) generated with MBP from both MS patients and normal controls. Here, we examined the frequency and peptide specificity of MBP-C8-specific TCL generated with MBP-C8 in MS patients and controls. Ten subjects grouped in five sets, each on MS patient and a control, were studied. In all cases, the MS patient had either a higher overall number of MBP-C8-responding lines, responded with greeter sensitivity to the MBP-C8 antigen or both. Few lines responded to the MBP-C8 peptides but if they did, they appeared to be specific to the carboxyl-half of the MBP-C8 molecule. Given the large amounts of citrullinated MBP in MS brain tissue, a preferential T cell response to MBP-C8 may be involved in the induction and perpetuation of this disease. C1 Univ Alabama, Dept Neurol, Birmingham, AL 35249 USA. NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Univ Alabama, Ctr Neuroimmunol, Birmingham, AL 35294 USA. Birmingham Vet Med Ctr, Neurol Serv, Birmingham, AL 35233 USA. Birmingham Vet Med Ctr, Res Serv, Birmingham, AL 35233 USA. Neurocrine Biosci San Diego, San Diego, CA 92121 USA. Univ Tubingen, Sch Med, Inst Physiol Chem, D-72076 Tubingen, Germany. RP Whitaker, JN (reprint author), Univ Alabama, Dept Neurol, 619 19th St S,JT 1205, Birmingham, AL 35249 USA. NR 26 TC 48 Z9 48 U1 0 U2 1 PU SAGE PUBLICATIONS LTD PI LONDON PA 1 OLIVERS YARD, 55 CITY ROAD, LONDON EC1Y 1SP, ENGLAND SN 1352-4585 J9 MULT SCLER J JI Mult. Scler. J. PD AUG PY 2000 VL 6 IS 4 BP 220 EP 225 PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 345XY UT WOS:000088841600002 PM 10962541 ER PT J AU Syed, NA Sandbrink, F Luciano, CA Altarescu, G Weibel, T Schiffmann, R Floeter, MK AF Syed, NA Sandbrink, F Luciano, CA Altarescu, G Weibel, T Schiffmann, R Floeter, MK TI Cutaneous silent periods in patients with Fabry disease SO MUSCLE & NERVE LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Association-of-Electrodiagnostic-Medicine CY OCT, 1999 CL VANCOUVER, CANADA SP Amer Assoc Electrodiagnost Med DE Fabry disease; quantitative sensory testing; reflexes; silent period; small fiber neuropathy; spinal circuitry ID MOTOR-NEURON EXCITABILITY; ALPHA-GALACTOSIDASE; STIMULATION; ABNORMALITIES; RESPONSES; REFLEX AB We assessed the cutaneous silent period (CSP) in 24 patients with fabry disease with small-fiber sensory neuropathy and 12 normal subjects to test the hypothesis that small-diameter afferents are responsible for producing the CSP. Sensory nerve conduction studies and quantitative sensory testing for cold and vibration detection thresholds were also measured. Overall, Fabry patients had impaired thermal, but not vibration, detection thresholds, with greatest impairment in the feet. In the upper extremity, CSP latencies, duration, and suppression of electromyographic activity (EMG) did not differ. In the lower extremity, patients had reduced suppression of EMG during the CSP compared to normal controls. CSP durations exhibited a bimodal distribution in patients, including a subset of seven patients with durations shorter than all controls. This subset had profound loss of thermal sensation in the feet, but this was also true of some patients who had normal CSPs, Patients with shortened CSPs had modestly elevated vibration thresholds and reduced sensory potentials in comparison to patients with normal CSPs. Reduced CSPs in Fabry patients are associated with, but not entirely explained by, the severity of small-fiber neuropathy as measured by quantitative sensory testing. The possibility that large-diameter fibers provide a minor contribution to producing the CSP should be considered. (C) 2000 John Wiley & Sons, Inc. C1 NINDS, EMG Sect, NIH, Bethesda, MD 20892 USA. RP Floeter, MK (reprint author), NINDS, EMG Sect, NIH, 10 Ctr Dr,MSC 1404, Bethesda, MD 20892 USA. NR 28 TC 30 Z9 35 U1 0 U2 0 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-639X J9 MUSCLE NERVE JI Muscle Nerve PD AUG PY 2000 VL 23 IS 8 BP 1179 EP 1186 DI 10.1002/1097-4598(200008)23:8<1179::AID-MUS4>3.0.CO;2-7 PG 8 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 338AE UT WOS:000088395300004 PM 10918253 ER PT J AU de Angelis, MH Flaswinkel, H Fuchs, H Rathkolb, B Soewarto, D Marschall, S Heffner, S Pargent, W Wuensch, K Jung, M Reis, A Richter, T Alessandrini, F Jakob, T Fuchs, E Kolb, H Kremmer, E Schaeble, K Rollinski, B Roscher, A Peters, C Meitinger, T Strom, T Steckler, T Holsboer, F Klopstock, T Gekeler, F Schindewolf, C Jung, T Avraham, K Behrendt, H Ring, J Zimmer, A Schughart, K Pfeffer, K Wolf, E Balling, R AF de Angelis, MH Flaswinkel, H Fuchs, H Rathkolb, B Soewarto, D Marschall, S Heffner, S Pargent, W Wuensch, K Jung, M Reis, A Richter, T Alessandrini, F Jakob, T Fuchs, E Kolb, H Kremmer, E Schaeble, K Rollinski, B Roscher, A Peters, C Meitinger, T Strom, T Steckler, T Holsboer, F Klopstock, T Gekeler, F Schindewolf, C Jung, T Avraham, K Behrendt, H Ring, J Zimmer, A Schughart, K Pfeffer, K Wolf, E Balling, R TI Genome-wide, large-scale production of mutant mice by ENU mutagenesis SO NATURE GENETICS LA English DT Article ID MOUSE; SPERMATOZOA AB In the post-genome era, the mouse will have a major role as a model system for functional genome analysis. This requires a large number of mutants similar to the collections available from other model organisms such as Drosophila melanogaster and Caenorhabditis elegans. Here we report on a systematic, genome-wide, mutagenesis screen in mice. As part of the German Human Genome Project, we have undertaken a large-scale ENU-mutagenesis screen for dominant mutations and a limited screen for recessive mutations'. In screening over 14,000 mice for a large number of clinically relevant parameters, we recovered 182 mouse mutants for a variety of phenotypes. In addition, 247 variant mouse mutants are currently in genetic confirmation testing and will result in additional new mutant lines. This mutagenesis screen, along with the screen described in the accompanying paper(2), leads to a significant increase in the number of mouse models(3) available to the scientific community. Our mutant lines are freely accessible to non-commercial users (for information, see http://www.gsf.de/ieg/groups/enu-mouse.html). C1 GSF, Res Ctr Environm & Hlth, Inst Expt Genet, Neuherberg, Germany. GSF, Res Ctr Environm & Hlth, Inst Pathol, Neuherberg, Germany. GSF, Res Ctr Environm & Hlth, Inst Immunol, Neuherberg, Germany. GSF, Res Ctr Environm & Hlth, Inst Mammalian Genet, Neuherberg, Germany. Tech Univ Munich, Inst Med Microbiol Immunol & Hyg, Munich, Germany. Univ Munich, Gene Ctr, Inst Mol Anim Breeding, Munich, Germany. Ingenium Pharmaceut AG, Martinsried, Germany. Max Delbrueck Ctr, Mol Genet & Mikrosatellitenzentrum, Berlin, Germany. GSF Munich, TUM, Div Environm Dermatol & Allergol, Munich, Germany. Clin Harlaching, Inst Clin Chem, Munich, Germany. Dr Von Haunerschen Kinderspital, Kinderklin, Abt Klin Chem Biochem & Stoffwechsel, Munich, Germany. Dr Von Haunerschen Kinderspital, Kinderpoliklin, Munich, Germany. Univ Klin Freiburg, Freiburg, Germany. Univ Munich, Klinikum Grosshadern, Dept Neurobiol, Munich, Germany. Univ Munich, Kinderklin, Abt Med Genet, Munich, Germany. Max Planck Inst Psychiat, D-80804 Munich, Germany. Bundesamt Strahlenschutz, Inst Strahlenhyg, Oberschleissheim, Germany. Tel Aviv Univ, Sackler Sch Med, Dept Human Genet, Tel Aviv, Israel. NIMH, Genet Lab, Bethesda, MD USA. Transgene SA, Strasbourg, France. RP de Angelis, MH (reprint author), GSF, Res Ctr Environm & Hlth, Inst Expt Genet, Neuherberg, Germany. EM hrabe@gsf.de RI Zimmer, Andreas/B-8357-2009; Jakob, Thilo/J-1621-2012; Rathkolb, Birgit/F-7041-2013; Fuchs, Helmut/M-7347-2014; Meitinger, Thomas/O-1318-2015; Hrabe de Angelis, Martin/F-5531-2012; Reis, Andre/D-2309-2009 OI Fuchs, Helmut/0000-0002-5143-2677; Hrabe de Angelis, Martin/0000-0002-7898-2353; Reis, Andre/0000-0002-6301-6363 NR 13 TC 438 Z9 444 U1 2 U2 16 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD AUG PY 2000 VL 25 IS 4 BP 444 EP 447 DI 10.1038/78146 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 341WY UT WOS:000088615000023 PM 10932192 ER PT J AU Yoshimoto, T Mizutani, H Tsutsui, H Noben-Trauth, N Yamanaka, K Tanaka, M Izumi, S Okamura, H Paul, WE Nakanishi, K AF Yoshimoto, T Mizutani, H Tsutsui, H Noben-Trauth, N Yamanaka, K Tanaka, M Izumi, S Okamura, H Paul, WE Nakanishi, K TI IL-18 induction of IgE: dependence on CD4(+) T cells, IL-4 and STAT6 SO NATURE IMMUNOLOGY LA English DT Article ID GAMMA-INDUCING FACTOR; B-CELLS; TH2 CELLS; INTERLEUKIN-18; RECEPTOR; CYTOKINE; LYMPHOKINE; EXPRESSION; INVIVO; ACTIVATION AB Overproduction of immunoglobulin E (IgE) and T helper cell type 2 (T(H)2) cytokines, including interleukin 4 (IL-4), IL-5 and IL-13, can result in allergic disorders. Although it is known that IL-4 is critical to the polarization of naive CD4(+) T cells to a T(H)2 phenotype, both in vitro and in many in vivo systems, other factors that regulate in vivo IL-4 production and T(H)2 commitment are poorly understood. IL-18, an IL-1-like cytokine that requires cleavage with caspase-1 to become active, was found to increase IgE production in a CD4(+) T cells-, IL-4- and STAT6-dependent fashion. IL-18 and T cell receptor-mediated stimulation could induce naive CD4(+) T cells to develop into IL-4-producing cells in vitro. Thus, caspase-1 and IL-18 may be critical in regulation of IgE production in vivo, providing a potential therapeutic target for allergic disorders. C1 Hyogo Coll Med, Inst Adv Med Sci, Dept Immunol & Med Zool, Nishinomiya, Hyogo 6638501, Japan. Hyogo Coll Med, Inst Adv Med Sci, Host Def Lab, Nishinomiya, Hyogo 6638501, Japan. Mie Univ, Fac Med, Dept Dermatol, Tsu, Mie 5148507, Japan. Mie Univ, Fac Med, Dept Biochem, Tsu, Mie 5148507, Japan. Natl Leprosarium, Kagawa 7610198, Japan. NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. Japan Sci & Technol Corp, Core Res Evolut Sci & Technol, Tokyo, Japan. RP Nakanishi, K (reprint author), Hyogo Coll Med, Inst Adv Med Sci, Dept Immunol & Med Zool, Nishinomiya, Hyogo 6638501, Japan. OI Tsutsui, Hiroko/0000-0001-7928-4875 NR 34 TC 230 Z9 237 U1 0 U2 0 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 2000 VL 1 IS 2 BP 132 EP 137 DI 10.1038/77811 PG 6 WC Immunology SC Immunology GA 363BW UT WOS:000089814900013 PM 11248805 ER PT J AU Herblot, S Steff, AM Hugo, P Aplan, PD Hoang, T AF Herblot, S Steff, AM Hugo, P Aplan, PD Hoang, T TI SCL and LMOI alter thymocyte differentiation: inhibition of E2A-HEB function and pre-T alpha chain expression SO NATURE IMMUNOLOGY LA English DT Article ID ACUTE LYMPHOBLASTIC-LEUKEMIA; TRANSCRIPTION FACTOR SCL; CELL DEVELOPMENT; TRANSGENIC MICE; LYMPHOCYTE DEVELOPMENT; B-LYMPHOCYTE; PROTEIN LMO2; GENE; E2A; ENHANCER AB Cooperation between the stem cell leukemia (SCL) transcription factor and its nuclear partners LMO1 or LMO2 induces aggressive T cell acute lymphoblastic leukemia when inappropriately expressed in T cells. This study examined the cellular and molecular targets of the SCL-LMO complex at the preleukemic stage. We show that SCL and its partners are coexpressed in the most primitive thymocytes. Maturation to the pre-T cell stage is associated with a down-regulation of SCL and LMO1 and LMO2, and a concomitant up-regulation of E2A and HEB expression. Moreover, enforced expression of SCL-LMO1 inhibits T cell differentiation and recapitulates a loss of HEB function, causing a deregulation of the transition checkpoint from the CD4(-)CD8(-) to CD4(+)CD8(+) stages. Finally, we identify the gene encoding pT alpha as a downstream target of HEB that is specifically repressed by the SCL-LMO complex. C1 Clin Res Inst Montreal, Montreal, PQ H2W 1R7, Canada. Procrea Biosci Inc, Montreal, PQ H4P 2R2, Canada. NCI, Dept Genet, Med Branch, Div Clin Sci, Gaithersburg, MD 20877 USA. Univ Montreal, Dept Pharmacol, Montreal, PQ H3C 3J7, Canada. Univ Montreal, Dept Biochem, Montreal, PQ H3C 3J7, Canada. Univ Montreal, Program Mol Biol, Montreal, PQ H3C 3J7, Canada. RP Hoang, T (reprint author), Clin Res Inst Montreal, 110 Pine Ave W, Montreal, PQ H2W 1R7, Canada. RI Aplan, Peter/K-9064-2016 NR 50 TC 154 Z9 155 U1 0 U2 2 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 2000 VL 1 IS 2 BP 138 EP 144 DI 10.1038/77819 PG 7 WC Immunology SC Immunology GA 363BW UT WOS:000089814900014 PM 11248806 ER PT J AU Anderson, HA Hiltbold, EM Roche, PA AF Anderson, HA Hiltbold, EM Roche, PA TI Concentration of MHC class II molecules in lipid rafts facilitates antigen presentation SO NATURE IMMUNOLOGY LA English DT Article ID T-CELL ACTIVATION; INFLUENZA-VIRUS HEMAGGLUTININ; GPI-ANCHORED PROTEINS; MEMBRANE DOMAINS; PEPTIDE COMPLEXES; CAVEOLAE MEMBRANE; HIGH-AFFINITY; RECEPTOR; MICRODOMAINS; CHOLESTEROL AB The plasma membranes of eukaryotic cells are not uniform and possess distinct cholesterol- and sphingolipid-rich raft microdomains that are enriched in proteins known to be essential for cellular function. Lipid raft microdomains are important for T cell receptor (TCR)-mediated activation of T cells. However, the importance of lipid rafts on antigen presenting cells (APCs) and their role in major histocompatibility (MHC) class II-restricted antigen presentation has not been examined. MHC class II molecules were found to be constitutively present in plasma membrane lipid rafts in B cells. Disruption of these microdomains dramatically inhibited antigen presentation at limiting concentrations of antigen. The inhibitory effect of raft disruption on antigen presentation could be overcome by loading the APCs with exceptionally high doses of antigen, showing that raft association concentrates MHC class II molecules into microdomains that allow efficient antigen presentation at low ligand densities. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Roche, PA (reprint author), NCI, Expt Immunol Branch, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 41 TC 254 Z9 256 U1 4 U2 10 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 2000 VL 1 IS 2 BP 156 EP 162 DI 10.1038/77842 PG 7 WC Immunology SC Immunology GA 363BW UT WOS:000089814900017 PM 11248809 ER PT J AU Watzl, C Long, EO AF Watzl, C Long, EO TI Exposing tumor cells to killer cell attack SO NATURE MEDICINE LA English DT Editorial Material ID T-CELLS; RECOGNITION; NKG2D; MICA AB Natural killer (NK) cells attack tumor and virally infected cells in the absence of antigen presentation, utilizing a combination of signals from activation and inhibitory receptors. Recent reports have identified the NK and T-cell surface protein NKG2D as a receptor for tumor cell ligands that activates killing of tumor targets. C1 NIAID, Immunogenet Lab, NIH, Rockville, MD 20852 USA. RP Watzl, C (reprint author), NIAID, Immunogenet Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. RI Long, Eric/G-5475-2011; Watzl, Carsten/B-4911-2013 OI Long, Eric/0000-0002-7793-3728; Watzl, Carsten/0000-0001-5195-0995 NR 9 TC 12 Z9 12 U1 0 U2 0 PU NATURE AMERICA INC 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 2000 VL 6 IS 8 BP 867 EP 868 DI 10.1038/78624 PG 2 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 376TX UT WOS:000165473800023 PM 10932221 ER PT J AU Yang, ZY Duckers, HJ Sullivan, NJ Sanchez, A Nabel, EG Nabel, GJ AF Yang, ZY Duckers, HJ Sullivan, NJ Sanchez, A Nabel, EG Nabel, GJ TI Identification of the Ebola virus glycoprotein as the main viral determinant of vascular cell cytotoxicity and injury SO NATURE MEDICINE LA English DT Article ID TRANSIENT TRANSFECTION; INFECTION AB Here we defined the main viral determinant of Ebola virus pathogenicity; synthesis of the virion glycoprotein (CP) of Ebola virus Zaire induced cytotoxic effects in human endothelial cells in vitro and in vivo. This effect mapped to a serine-threonine-rich, mucin-like domain of this type I transmembrane glycoprotein, one of seven gene products of the virus. Gene transfer of GP into explanted human or porcine blood vessels caused massive endothelial cell loss within 48 hours that led to a substantial increase in vascular permeability. Deletion of the mucin-like region of GP abolished these effects without affecting protein expression or function. CP derived from the Reston strain of virus, which causes disease in nonhuman primates but not in man, did not disrupt the vasculature of human blood vessels. In contrast, the Zaire CP induced endothelial cell disruption and cytotoxicity in both nonhuman primate and human blood vessels, and the mucin domain was required for this effect. These findings indicate that CP, through its mucin domain, is the viral determinant of Ebola pathogenicity and likely contributes to hemorrhage during infection. C1 NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. NHLBI, Vasc Biol Branch, NIH, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Special Pathogens Branch, Div Viral & Rickettsial Dis, Natl Ctr Infect Dis, Atlanta, GA 30333 USA. RP Nabel, GJ (reprint author), NIH, Vaccine Res Ctr, 40 Convent Dr, Bethesda, MD 20892 USA. NR 18 TC 244 Z9 263 U1 2 U2 52 PU NATURE AMERICA INC 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 2000 VL 6 IS 8 BP 886 EP 889 PG 4 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 376TX UT WOS:000165473800031 PM 10932225 ER PT J AU Atzori, M Lau, D Tansey, EP Chow, A Ozaita, A Rudy, B McBain, CJ AF Atzori, M Lau, D Tansey, EP Chow, A Ozaita, A Rudy, B McBain, CJ TI H-2 histamine receptor-phosphorylation of Kv3.2 modulates interneuron fast spiking SO NATURE NEUROSCIENCE LA English DT Article ID HIPPOCAMPAL PYRAMIDAL CELLS; CHANNEL SUBUNIT KV3.1B; HIGH-FREQUENCY OSCILLATIONS; GATED K+ CHANNELS; POTASSIUM-CHANNEL; RAT HIPPOCAMPUS; NEOCORTICAL INTERNEURONS; DIFFERENTIAL EXPRESSION; BEHAVING RAT; CNS NEURONS AB Histamine-containing neurons of the tuberomammilary nucleus project to the hippocampal formation to innervate H-1 and H-2 receptors on both principal and inhibitory interneurons. Here we show that H-2 receptor activation negatively modulates outward currents through Kv3.2-containing potassium channels by a mechanism involving PKA phosphorylation in inhibitory interneurons. PKA phosphorylation of Kv3.2 lowered the maximum firing frequency of inhibitory neurons, which in turn negatively modulated high-frequency population oscillations recorded in principal cell layers. All these effects were absent in a Kv3.2 knockout mouse. These data reveal a novel pathway for histamine-dependent regulation of high-frequency oscillations within the hippocampal formation. C1 NICHD, Lab Cellular & Mol Neurophysiol, NIH, Bethesda, MD 20892 USA. NYU, Sch Med, Dept Physiol & Neurosci, New York, NY 10016 USA. NYU, Sch Med, Dept Biochem, New York, NY 10016 USA. RP McBain, CJ (reprint author), NICHD, Lab Cellular & Mol Neurophysiol, NIH, Rm 5A72,Bldg 49,Convent Dr, Bethesda, MD 20892 USA. FU NINDS NIH HHS [NS30989, NS35215] NR 48 TC 67 Z9 70 U1 0 U2 3 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 2000 VL 3 IS 8 BP 791 EP 798 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 405TU UT WOS:000167177300014 PM 10903572 ER PT J AU Turkheimer, F Pettigrew, K Sokoloff, L Smith, CB Schmidt, K AF Turkheimer, F Pettigrew, K Sokoloff, L Smith, CB Schmidt, K TI Selection of an adaptive test statistic for use with multiple comparison analyses of neuroimaging data SO NEUROIMAGE LA English DT Article; Proceedings Paper CT XVIIIth International Symposium on Cerebral Blood Flow and Metabolism CY JUN 15-19, 1997 CL BALTIMORE, MARYLAND DE permutation tests; randomization tests; multiple comparisons; autoradiography; positron emission tomography (PET) ID RANDOMIZATION TEST AB Statistical analysis of neuroimages is commonly approached with intergroup comparisons made by repeated application of univariate or multivariate tests performed on the set of the regions of interest sampled in the acquired images. The use of such large numbers of tests requires application of techniques for correction for multiple comparisons. Standard multiple comparison adjustments (such as the Bonferroni) may be overly conservative when data are correlated and/or not normally distributed. Resampling-based step-down procedures that successfully account for unknown correlation structures in the data have recently been introduced. We combined resampling step-down procedures with the Minimum Variance Adaptive method, which allows selection of an optimal test statistic from a predefined class of statistics for the data under analysis. As shown in simulation studies and analysis of autoradiographic data, the combined technique exhibits a significant increase in statistical power, even for small sample sizes (n = 8, 9, 10). (C) 2000 Academic Press. C1 NIMH, Cerebral Metab Lab, NIH, Bethesda, MD 20892 USA. NIMH, Div Epidemiol & Serv Res, NIH, Bethesda, MD 20892 USA. RP Schmidt, K (reprint author), NIMH, Cerebral Metab Lab, NIH, Bldg 36,Room 1A-07,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Turkheimer, Federico/B-9485-2012 OI Turkheimer, Federico/0000-0002-3766-3815 NR 39 TC 4 Z9 4 U1 0 U2 3 PU ACADEMIC PRESS INC 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 2000 VL 12 IS 2 BP 219 EP 229 DI 10.1006/nimg.2000.0608 PG 11 WC Neurosciences; Neuroimaging; Radiology, Nuclear Medicine & Medical Imaging SC Neurosciences & Neurology; Radiology, Nuclear Medicine & Medical Imaging GA 343FZ UT WOS:000088692200009 PM 10913327 ER PT J AU Erb, S Hitchcott, PK Rajabi, H Mueller, D Shaham, Y Stewart, J AF Erb, S Hitchcott, PK Rajabi, H Mueller, D Shaham, Y Stewart, J TI Alpha-2 adrenergic receptor agonists block stress-induced reinstatement of cocaine seeking SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE clonidine; lofexidine; cocaine self-administration; relapse; stress; microdialysis ID CORTICOTROPIN-RELEASING-FACTOR; SELF-ADMINISTRATION BEHAVIOR; VENTRAL TEGMENTAL AREA; FREELY-MOVING RATS; INDUCED RELAPSE; LOCUS-CERULEUS; NORADRENALINE RELEASE; NORADRENERGIC NEURONS; HEMODYNAMIC STRESS; CLONIDINE ANALOGS AB The alpha-2 adrenergic receptor agonists, cloindine, lofexidine and guanabenz, blocked stress- but not cocaine-induced reinstatement of cocaine seeking at doses that suppressed footshock-induced release of noradrenaline in prefrontal cortex and amygdala. Rats were trained to self-administer cocaine (0.5 mg/kg/infusion, i.v; 10-12 days) and, after a drug-free period (7-13 days), were returned to the self-administration chambers for daily extinction and reinstatement test sessions. Both intermittent footshock (15 min, 0.6 IIIA) and cocaine priming (20 mg/kg, i.p.) reinstated extinguished drug seeking. Pretreatment with either clonidine (20, or 40 mu g/kg, i.p.) or lofexidine (50, 100, 150, or 200 mu g/kg, i.p.) attenuated footshock- but not cocaine-induced reinstatement of cocaine seeking. Guanabenz (640 mu g/kg, ip) an alpha-2 agonist with low affinity for imidazoline type-1 receptors, also attenuated footshock- but but not cocaine-induced reinstatement of cocaine seeking. The results point to an important role for NE systems in the effects of footshock on relapse to cocaine seeking. (C) 2000 American College of Neuropsychopharmacology. Published by Elsevier Science Inc. All rights reserved. C1 Concordia Univ, Ctr Studies Behav Neurobiol, Dept Psychol, Montreal, PQ H3G 1M8, Canada. NIDA, IRP, Dept Behav Neurosci, Baltimore, MD USA. RP Stewart, J (reprint author), Concordia Univ, Ctr Studies Behav Neurobiol, Dept Psychol, 1455 Maisonneuve Blvd W, Montreal, PQ H3G 1M8, Canada. RI shaham, yavin/G-1306-2014; OI Mueller, Devin/0000-0002-7397-9219 NR 61 TC 177 Z9 181 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD AUG PY 2000 VL 23 IS 2 BP 138 EP 150 DI 10.1016/S0893-133X(99)00158-X PG 13 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 335MC UT WOS:000088246400003 PM 10882840 ER PT J AU Castro-Caldas, A Grafman, J AF Castro-Caldas, A Grafman, J TI Those were the (phrenological) days SO NEUROSCIENTIST LA English DT Article DE phrenology; Franz Joseph Gall; Johann Spurzheim; history of neurosciences AB Phrenology is nowadays often considered as a pseudoscience and a reason for ironic comments. This theory was based on the ideas of Franz Joseph Gall (1758-1828). He was indeed a fine neuroscientist that called attention to the cerebral cortex and suggested for the first time that mental functions were the result of the activity of cortical "organs." The methodology used reveals interesting predictive aspects for what our current practice is. However, the interpretations of the observations were often more in accordance with the predicted heuristic model than with the real findings. Spurzheim (1776-1832) was first his student and then his collaborator. He traveled around the world lecturing and making phrenology a popular science. C1 Univ Lisbon, Hosp Santa Maria, Dept Neurol, P-1649035 Lisbon, Portugal. NINDS, Cognit Neurosci Sect, Bethesda, MD 20892 USA. OI Grafman, Jordan H./0000-0001-8645-4457; Castro-Caldas, Alexandre/0000-0002-9148-3719 NR 15 TC 2 Z9 2 U1 2 U2 4 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1073-8584 J9 NEUROSCIENTIST JI Neuroscientist PD AUG PY 2000 VL 6 IS 4 BP 297 EP 302 PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 339FE UT WOS:000088466300021 ER PT J AU Matveeva, OV Tsodikov, AD Giddings, M Freier, SM Wyatt, JR Spiridonov, AN Shabalina, SA Gesteland, RF Atkins, JF AF Matveeva, OV Tsodikov, AD Giddings, M Freier, SM Wyatt, JR Spiridonov, AN Shabalina, SA Gesteland, RF Atkins, JF TI Identification of sequence motifs in oligonucleotides whose presence is correlated with antisense activity SO NUCLEIC ACIDS RESEARCH LA English DT Article ID EXPRESSION; INHIBITION; PHOSPHOROTHIOATE; CELLS; RNA; KINASE; OLIGODEOXYNUCLEOTIDE; DUPLEXES; ICAM-1; ALPHA AB Design of antisense oligonucleotides targeting any mRNA can be much more efficient when several activity-enhancing motifs are included and activity-decreasing motifs are avoided, This conclusion was made after statistical analysis of data collected from >1000 experiments with phosphorothioate-modified oligonucleotides. Highly significant positive correlation between the presence of motifs CCAC, TCCC, ACTC, GCCA and CTCT in the oligonucleotide and its antisense efficiency was demonstrated, In addition, negative correlation was revealed for the motifs GGGG, ACTG, AAA and TAA, It was found that the likelihood of activity of an oligonucleotide against a desired mRNA target is sequence motif content dependent. C1 Univ Utah, Dept Human Genet, Salt Lake City, UT 84112 USA. Univ Utah, Dept Oncol Sci, Huntsman Canc Inst, Salt Lake City, UT 84112 USA. ISIS Pharmaceut, Carlsbad, CA 92008 USA. IHS, Ithaca, NY 14853 USA. NIH, Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20814 USA. RP Matveeva, OV (reprint author), Univ Utah, Dept Human Genet, 15N 2030E Room 7410, Salt Lake City, UT 84112 USA. RI Shabalina, Svetlana/N-8939-2013 OI Shabalina, Svetlana/0000-0003-2272-7473 FU NCI NIH HHS [2P30 CA42014-12, P30 CA042014]; NIGMS NIH HHS [GM 48152, R01 GM048152] NR 22 TC 47 Z9 53 U1 0 U2 2 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 AUG 1 PY 2000 VL 28 IS 15 BP 2862 EP 2865 DI 10.1093/nar/28.15.2862 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 340ZW UT WOS:000088567400002 PM 10908347 ER PT J AU Bank, MS Franklin, WL Sarno, RJ AF Bank, MS Franklin, WL Sarno, RJ TI Assessing the effect of radiocollars on juvenile guanaco survival SO OECOLOGIA LA English DT Article DE radiocollar; survival; guanaco; Lama guanicoe; ungulate ID IMMOBILIZATION; MORTALITY; MOOSE; CHILE AB To evaluate the potential impact of our research activities on guanacos (Lama guanicoe), we tested for effects of radiocollaring on juvenile (less than or equal to 1 year of age) survival in Torres del Paine National Park, Chile during 1992-1996. The survival of collared (40.1%) and uncollared (38.2%) juveniles was not significantly different (G = 0.08; P = 0.77). Our results suggest that radiocollaring does not adversely affect the survival of juvenile guanacos. Although we observed no effect of radiocollaring, we still underline the importance of testing for effects of radiocollars in other wildlife investigations when feasible. C1 Univ Maine, Dept Wildlife Ecol, Orono, ME 04469 USA. Iowa State Univ Sci & Technol, Dept Anim Ecol, Ames, IA 50011 USA. NCI, Lab Genom Divers, FCRDC, Ft Detrick, MD 21702 USA. RP Bank, MS (reprint author), Univ Maine, Dept Wildlife Ecol, 5755 Nutting Hall, Orono, ME 04469 USA. NR 22 TC 9 Z9 9 U1 2 U2 4 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0029-8549 J9 OECOLOGIA JI Oecologia PD AUG PY 2000 VL 124 IS 2 BP 232 EP 234 DI 10.1007/s004420050011 PG 3 WC Ecology SC Environmental Sciences & Ecology GA 345PJ UT WOS:000088824100011 PM 28308183 ER PT J AU McCabe, M Newsome, S Cheson, BD AF McCabe, M Newsome, S Cheson, BD TI Clinical trials - Referral resource SO ONCOLOGY-NEW YORK LA English DT Editorial Material C1 NCI, Bethesda, MD 20892 USA. RP McCabe, M (reprint author), NCI, Bethesda, MD 20892 USA. NR 0 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 2000 VL 14 IS 8 BP 1181 EP 1181 PG 1 WC Oncology SC Oncology GA 368QT UT WOS:000090129600014 ER PT J AU Lembo, T Naliboff, BD Matin, K Munakata, J Parker, RA Gracely, RH Mayer, EA AF Lembo, T Naliboff, BD Matin, K Munakata, J Parker, RA Gracely, RH Mayer, EA TI Irritable bowel syndrome patients show altered sensitivity to exogenous opioids SO PAIN LA English DT Article DE fentanyl; antinociception; visceral pain ID POSITRON EMISSION TOMOGRAPHY; HEART-RATE-VARIABILITY; OPIATE RECEPTORS; SPECTRAL-ANALYSIS; BRAIN-STEM; MORPHINE; PAIN; HYPERSENSITIVITY; HYPERALGESIA; PERCEPTION AB Alterations in activation of pain modulation systems may play a role in the pathophysiology of irritable bowel syndrome (IBS). However, little is known about the effects of exogenous opioids on the perceptual and autonomic responses to aversive visceral stimulation. The aim of the study was to evaluate the effect of the mu opioid-preferring analgesic fentanyl (FEN), given intravenously, on perceptual and autonomic responses to rectal distension. Ten IBS patients and ten normal subjects received, on separate days, either high dose (HD) fentanyl (112 mu g bolus followed by 0.04 mu g/kg per min infusion), low dose (L.D) fentanyl (56 mu g bolus followed by 0.02 mu g/kg per min) or normal saline (SAL) (50 cc bolus followed by 45 cc/h infusion). Perception thresholds for discomfort and pain during rectal distension were assessed using a tracking paradigm. Intensity and unpleasantness ratings of the distensions, and cardiac autonomic parameters were assessed during randomly delivered rectal stimuli. Effects of FEN on rectal compliance and tone as well as mental status were also assessed. IBS patients had lower perceptual thresholds for discomfort and pain under control conditions. FEN dose-dependently increased the perception thresholds in both healthy control subjects and in IBS patients with a greater relative efficacy in IUS patients than in normal subjects. IBS patients used significantly higher unpleasantness ratings of rectal stimuli compared to healthy controls, but showed no difference in the sensory intensity rating of the stimulus. FEN decreased both intensity and unpleasantness ratings for IBS and normals. FEN lowered cardiosympathetic tone in normal subjects but had no effect on IBS patients. FEN had no effect on rectal tone or compliance. FEN dose-dependently attenuates the perception of phasic rectal distension and affects unpleasantness ratings during random fixed rectal distension, with a greater relative efficacy for this antinociceptive effect in IBS patients. These findings support the hypothesis that IBS patients may have an altered central release of endogenous opioids in response to visceral stimulation. (C) 2000 International Association for the Study of Pain. Published by Elsevier Science B.V. All rights reserved. C1 Beth Israel Deaconess Med Ctr, Dept Med, Boston, MA USA. Univ Calif Los Angeles, W Los Angeles Vet Affairs Healthcare Med Ctr, Dept Psychiat & Biobehav Sci, Los Angeles, CA USA. Univ Calif Los Angeles, Sch Med, Dept Med, Neuroenter Dis Program,CURE Digest Dis Res Ctr, Los Angeles, CA USA. NIDCR, PNMB, CMMU, NIH, Bethesda, MD USA. Univ Calif Los Angeles, Sch Med, Dept Physiol, Los Angeles, CA 90024 USA. RP Mayer, EA (reprint author), Univ Calif Los Angeles, WLA VA Med Ctr, Div Digest Dis, Bldg 115 CURE,11301 Wilshire Blvd, Los Angeles, CA 90073 USA. EM emayer@ucla.edu NR 45 TC 76 Z9 76 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0304-3959 EI 1872-6623 J9 PAIN JI Pain PD AUG PY 2000 VL 87 IS 2 BP 137 EP 147 DI 10.1016/S0304-3959(00)00282-7 PG 11 WC Anesthesiology; Clinical Neurology; Neurosciences SC Anesthesiology; Neurosciences & Neurology GA 345KA UT WOS:000088814100004 PM 10924807 ER PT J AU Bustamante, PJ Woodruff, DC Oh, J Keister, DB Muratova, O Williamson, KC AF Bustamante, PJ Woodruff, DC Oh, J Keister, DB Muratova, O Williamson, KC TI Differential ability of specific regions of Plasmodium falciparum sexual-stage antigen, Pfs230, to induce malaria transmission-blocking immunity SO PARASITE IMMUNOLOGY LA English DT Article DE malaria; Plasmodium falciparum; vaccine; transmission-blocking; Pfs230 ID GAMETE SURFACE PROTEIN; ANTIBODIES; TARGET; RECOGNITION; PFS48/45; VACCINE; SERA; GENE AB Antibodies raised against an Escherichia coli-produced recombinant protein encoding a 76-kDa section (region C) of malaria transmission-blocking vaccine candidate, Pfs230, have previously been shown to significantly reduce the ability of Plasmodium falciparum parasites to infect mosquitoes (71.2-89.8%). To further define the region of the Pfs230 required for transmission-blocking activity, four recombinant proteins each encoding a section of region C (Pfs230 amino acids 443-1132) were produced using the same E. coli expression system and tested for immogenicity in mice: (i) r230/MBP.C5' encodes the first half of region C (amino acids 443-791, six cysteines); (ii) r230/MBP.CM1 encodes only cysteine motif (CM) 1 (amino acids 583-913, eight cysteines); (iii) r230/MBP.C1.6 (amino acids 453-913, eight cysteines) also includes all of CM1; and (iv) r230/MBP.C2 encodes only CM2 (amino acids 914-1268, 11 cysteines). All the recombinant proteins induced antibodies that recognized parasite-produced Pfs230, but the titre of the Pfs230 specific-antibodies generated varied, C = C1.6 = C5' > CM1 > CM2. Two recombinants, r230/MBP.C5' and r230/MBP.C1.6, induced antibody titres that were equivalent to or greater than the titre generated by r230/MBP.C. However, in contrast to r230/MBP.C, none of the recombinants induced antibodies that effectively blocked parasite infectivity to mosquitoes. This suggests that the inclusion of amino acids 914-1132 is important for the production of the transmission-blocking epitope present in region C. C1 Loyola Univ, Dept Biol, Chicago, IL 60626 USA. NIAID, Malaria Vaccine Dev Unit, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Williamson, KC (reprint author), Loyola Univ, Dept Biol, 6525 N Sheridan Rd, Chicago, IL 60626 USA. NR 16 TC 24 Z9 26 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0141-9838 J9 PARASITE IMMUNOL JI Parasite Immunol. PD AUG PY 2000 VL 22 IS 8 BP 373 EP 380 PG 8 WC Immunology; Parasitology SC Immunology; Parasitology GA 343HP UT WOS:000088697200001 PM 10972844 ER PT J AU Chiou, CC Groll, AH Gonzalez, CE Callender, D Venzon, D Pizzo, PA Wood, L Walsh, TJ AF Chiou, CC Groll, AH Gonzalez, CE Callender, D Venzon, D Pizzo, PA Wood, L Walsh, TJ TI Esophageal candidiasis in pediatric acquired immunodeficiency syndrome: clinical manifestations and risk factors SO PEDIATRIC INFECTIOUS DISEASE JOURNAL LA English DT Article; Proceedings Paper CT 37th Annual Meeting of the Infectious-Diseases-Society-of-America CY NOV 18-21, 1999 CL PHILADELPHIA, PENNSYLVANIA SP Infect Dis Soc Amer DE Candidiasis; esophagitis; pediatric acquired immunodeficiency; syndrome; Candida albicans; risk factors ID VIRUS-INFECTED PATIENTS; ENDOSCOPY; OROPHARYNGEAL; DIAGNOSIS; COLONIZATION; FLUCONAZOLE; CANDIDEMIA; ALBICANS; MUCOSAL; CANCER AB Background. Little is known about the epidemiology and clinical features of esophageal candidiasis (EC) in pediatric AIDS. We therefore investigated the clinical presentation and risk factors of EC in a large prospectively monitored population of HIV-infected children at the National Cancer Institute. Patients and methods. We reviewed the records of all HIV-infected children (N = 448) followed between 1987 and 1995 for a history of esophageal candidiasis to characterize the epidemiology, clinical features, therapeutic interventions and outcome of esophageal candidiasis. To understand further the risk factors for EC in pediatric AIDS, we then performed a matched case-control analysis of 25 patients for whom control cases were available. Results. There were 51 episodes of EC documented in 36 patients with 23 male and 13 female patients (0.2 to 17 years; median CD4, count 11/mu l), representing a frequency of EC of 8.0%, Concurrent oropharyngeal candidiasis (OPC) was the most common clinical presentation of EC (94%); other signs and symptoms included odynophagia (80%), retrosternal pain (57%), fever (29%), nausea/vomiting (24%), drooling (12%), dehydration (12%), hoarseness (6%) and upper gastrointestinal bleeding (6%), The causative organism documented in 36 episodes (18 from OPC, 17 from endoscopic biopsy and 1 from autopsy) was Candida albicans in all cases, Patients received treatment for EC with amphotericin B (63%), fluconazole (29%), ketoconazole (4%) or itraconazole (1%), A clinical response was documented in all 45 evaluable episodes. In 6 other cases, EC was a final event without contributing to the cause of death, By a conditional logistic regression model for matched data, the best predictor of EC was the presence of prior OPC (P < 0.0001), followed by CD4 count and CD4 percentage (P = 0.0002) and use of antibacterial antibiotics (P = 0.0013), The risks associated with low CD4 count were independent of that of prior OPC. Conclusion, EC in pediatric AIDS is a debilitating infection, which develops in the setting of prior OPC, low CD4 counts and previous antibiotics. C1 NCI, Immunocompromised Host Sect, Pediat Oncol Branch, Bethesda, MD 20892 USA. NCI, HIV & AIDS Related Malignancy Branch, Bethesda, MD 20892 USA. NCI, Biostat & Data Management Sect, Bethesda, MD 20892 USA. NCI, Pediat Branch, Bethesda, MD 20892 USA. RP Walsh, TJ (reprint author), NCI, Immunocompromised Host Sect, Pediat Oncol Branch, Bldg 10,Rm 13N-240, Bethesda, MD 20892 USA. RI Venzon, David/B-3078-2008 NR 33 TC 19 Z9 21 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0891-3668 J9 PEDIATR INFECT DIS J JI Pediatr. Infect. Dis. J. PD AUG PY 2000 VL 19 IS 8 BP 729 EP 734 DI 10.1097/00006454-200008000-00011 PG 6 WC Immunology; Infectious Diseases; Pediatrics SC Immunology; Infectious Diseases; Pediatrics GA 344JE UT WOS:000088754800010 PM 10959741 ER PT J AU Izevbigie, EB Gutkind, JS Ray, PE AF Izevbigie, EB Gutkind, JS Ray, PE TI Isoproterenol inhibits fibroblast growth factor-2-induced growth of renal epithelial cells SO PEDIATRIC NEPHROLOGY LA English DT Article DE basic fibroblast growth factor; isoproterenol; renal tubular epithelial cells; cAMP; mitogen-activated protein kinases; MEK 1/2; cell proliferation ID ACTIVATED PROTEIN-KINASE; HIV-ASSOCIATED NEPHROPATHY; MAP KINASE; DEPENDENT ACTIVATION; ADRENERGIC-RECEPTORS; SIGNAL-TRANSDUCTION; MITOGENIC RESPONSE; KIDNEY; CAMP; PATHWAY AB The signal transduction pathways modulating bFGF effects in renal tubular epithelial cells (RTEc) are not completely understood. Since the cAMP and the mitogen-activated protein kinase (MAPK) pathways can modulate the growth of RTEc, we studied whether two cAMP elevating agents, isoproterenol and 8-bromo-cAMP, would modulate basic fibroblast growth factor (bFGF) induction of MAPK activity (ERK-2) and cell proliferation in human renal proximal tubular epithelial cells (RPTEc) and Madin-Darby canine kidney cells (MDCK clone (E11)). Isoproterenol, but not bFGF, stimulated cAMP production in RPTEc and MDCKE11 cells. bFGF, isoproterenol, and 8-bromo-cAMP alone increased ERK-2 activity in both cell types. However, isoproterenol and 8-bromo-cAMP partially inhibited the bFGF induction of ERK-2 activity, but only isoproterenol inhibited the proliferation of both cell types. PD098059 (25 mu M). an inhibitor of MAPK kinase (MEK 1/2). blocked the bFGF mitogenic effects, but did not affect the 8-bromo-cAMP-induced mitogenic effects in MDCKE11 cells. These findings suggest that activation of ERK-2 is required but not sufficient for mitogenesis in RTEc. We conclude that isoproterenol inhibits the growth-promoting effects of bFGF in RTEc via MEK-dependent and -independent pathways. C1 George Washington Univ, Childrens Natl Med Ctr, Childrens Res Inst, Ctr Mol Physiol, Washington, DC 20010 USA. NIDR, Oral Pharyngeal Canc Branch Program, NIH, Bethesda, MD 20892 USA. RP Ray, PE (reprint author), George Washington Univ, Childrens Natl Med Ctr, Childrens Res Inst, Ctr Mol Physiol, Room R-211,3-5 Floor,111 Michigan Ave NW, Washington, DC 20010 USA. EM Pray@cnmc.org RI Gutkind, J. Silvio/A-1053-2009 FU NHLBI NIH HHS [R0-1HL 55605]; NIDDK NIH HHS [R0-1DK 4919, DK49419-S1] NR 42 TC 13 Z9 13 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0931-041X J9 PEDIATR NEPHROL JI Pediatr. Nephrol. PD AUG PY 2000 VL 14 IS 8-9 BP 726 EP 734 PG 9 WC Pediatrics; Urology & Nephrology SC Pediatrics; Urology & Nephrology GA 343HG UT WOS:000088695900006 PM 10955916 ER PT J AU Orvisky, E Sidransky, E McKinney, CE LaMarca, ME Samimi, R Krasnewich, D Martin, BM Ginns, EI AF Orvisky, E Sidransky, E McKinney, CE LaMarca, ME Samimi, R Krasnewich, D Martin, BM Ginns, EI TI Glucosylsphingosine accumulation in mice and patients with type 2 Gaucher disease begins early in gestation SO PEDIATRIC RESEARCH LA English DT Article ID GLOBOID-CELL LEUKODYSTROPHY; PSYCHOSINE; GENE AB Gaucher disease, the most common of the sphingolipidoses, results from the inherited deficiency of the enzyme glucocerebrosidase (EC 3.2.1.45). Although type 2 (acute neuronopathic) Gaucher disease is associated with rapidly progressive and fatal neurologic deterioration, the pathophysiologic mechanisms leading to the neurologic symptoms and early demise remain uncharacterized. While the pathology encountered in Gaucher disease has been attributed to glucocerebroside storage, glucosylsphingosine (Glc-sph), a cytotoxic compound, also accumulates in the tissues. Elevations of brain Glc-sph have been reported in patients with types 2 and 3 Gaucher disease. In this study, Glc-sph levels were measured using HPLC in tissues from mice with type 2 Gaucher disease created with a null glucocerebrosidase allele. Compared with unaffected littermates, homozygous mice with type 2 Gaucher disease had approximately a 100-fold elevation of Glc-sph in brain, as well as elevated levels in other tissues. This accumulation was detected in utero by E 13 and increased progressively throughout gestation. Similarly, elevated Glc-sph levels were seen in human fetuses with type 2 Gaucher disease, indicating that therapy initiated after birth may be too late to prevent the sequaelae of progressive neurologic damage that begins early in gestation. These findings suggest that the accumulation of Glc-sph may be responsible for the rapid demise of mice with type 2 Gaucher disease and the devastating clinical course seen in patients with type 2 Gaucher disease. C1 NIMH, Clin Neurosci Branch, NIH, Bethesda, MD 20892 USA. RP Sidransky, E (reprint author), NIMH, Clin Neurosci Branch, NIH, Bldg 49,Room B1EE16,49 Convent Dr,MSC4405, Bethesda, MD 20892 USA. NR 30 TC 56 Z9 56 U1 0 U2 3 PU INT PEDIATRIC RESEARCH FOUNDATION, INC PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 USA SN 0031-3998 J9 PEDIATR RES JI Pediatr. Res. PD AUG PY 2000 VL 48 IS 2 BP 233 EP 237 DI 10.1203/00006450-200008000-00018 PG 5 WC Pediatrics SC Pediatrics GA 339VX UT WOS:000088499500018 PM 10926300 ER PT J AU Brunell, PA Argaw, T AF Brunell, PA Argaw, T TI Chickenpox attributable to a vaccine virus contracted from a vaccinee with zoster SO PEDIATRICS LA English DT Article DE varicella-zoster; zoster; vaccine; transmission; rash; PstI ID ATTENUATED VARICELLA VACCINE; CHILDREN; LIVE; LEUKEMIA; TRANSMISSION AB Five months after 2 siblings were immunized with varicella vaccine, 1 developed zoster. Two weeks later the second sibling got a mild case of chicken pox. Virus isolated from the latter was found to be vaccine type. Thus, the vaccine strain was transmitted from the vaccinee with zoster to his sibling. Vaccinees who later develop zoster must be considered contagious. C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Cedars Sinai Med Ctr, Ahmanson Pediat Dept, Los Angeles, CA 90048 USA. RP Brunell, PA (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10,Room 11N228,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 14 TC 19 Z9 22 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 2000 VL 106 IS 2 BP art. no. EP e28 DI 10.1542/peds.106.2.e28 PG 2 WC Pediatrics SC Pediatrics GA 340MM UT WOS:000088538100014 PM 10920184 ER PT J AU Chase, C Ware, J Hittelman, J Blasini, I Smith, R Llorente, A Anisfeld, E Diaz, C Fowler, MG Moye, J Kaligh, LI AF Chase, C Ware, J Hittelman, J Blasini, I Smith, R Llorente, A Anisfeld, E Diaz, C Fowler, MG Moye, J Kaligh, LI CA Women Infants Transmission Study G TI Early cognitive and motor development among infants born to women infected with human immunodeficiency virus SO PEDIATRICS LA English DT Article DE human immunodeficiency virus; child development ID CHILDREN; NEURODEVELOPMENT; BEHAVIOR AB Objective. To examine the frequency, timing, and factors associated with abnormal cognitive and motor development during the first 30 months of life in infants born to women infected with human immunodeficiency virus type 1 (HIV-1). Methods. Serial neurodevelopmental assessment was performed with 595 infants born to women infected with HIV-1 in a multicenter, prospective, natural history cohort study. Survival analysis methods were used to evaluate 6 outcome events related to abnormal cognitive and motor growth (time to confirmed drop of 1 SD, time to first score <69, and time to confirmed drop of 2 SD) in Bayley Scales of Infant Development Mental Developmental Index (MDI) and Psychomotor Developmental Index (PDI) scores among infected (n = 114) and uninfected (n = 481) infants. Proportional hazards modeling was used to evaluate the effects of HIV infection status, prematurity, prenatal exposure to illicit drugs, maternal educational attainment, and primary language. Results. HIV-1 infection was significantly associated with increased risk for all outcome events related to abnormal mental and motor growth. Differences between infected and uninfected infants were apparent by 4 months of age. Prematurity was associated with increased risk for MDI <69 and PDI <69. Maternal education of <9 completed years was associated with increased risk for MDI <69. Neither prenatal exposure to illicit drugs nor primary language other than English was associated with abnormal development. Conclusion. A significant proportion of infants with HIV-1 infection show early and marked cognitive and motor delays or declines that may be important early indicators of HIV disease progression. These abnormalities are independent of other risk factors for developmental delay. C1 Boston Univ, Sch Med, Boston Med Ctr, Dept Pediat,Div Infect Dis, Boston, MA 02118 USA. Harvard Univ, Sch Med, Childrens Hosp, Dept Pediat,Div Infect Dis, Boston, MA USA. SUNY Hlth Sci Ctr, Brooklyn, NY 11203 USA. Univ Puerto Rico, Dept Pediat, San Juan, PR 00936 USA. Univ Illinois, Dept Pediat, Urbana, IL 61801 USA. Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. Columbia Presbyterian Hosp, Dept Pediat, New York, NY USA. NIAID, Div Acquired Immunodeficiency Syndrome, Bethesda, MD 20892 USA. NICHHD, Pediat Adolescent & Maternal AIDS Branch, Washington, DC USA. New England Res Inst, Watertown, MA 02172 USA. RP Chase, C (reprint author), Boston Univ, Sch Med, Boston Med Ctr, Dept Pediat,Div Infect Dis, DLG 7,1 Boston Med Ctr Pl, Boston, MA 02118 USA. FU NIAID NIH HHS [UO1 AI 34842, U01 AI34858, UO1 AI 34856] NR 21 TC 41 Z9 43 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 2000 VL 106 IS 2 AR e25 DI 10.1542/peds.106.2.e25 PG 10 WC Pediatrics SC Pediatrics GA 340MM UT WOS:000088538100011 PM 10920181 ER PT J AU Trobst, KK AF Trobst, KK TI An interpersonal conceptualization and quantification of social support transactions SO PERSONALITY AND SOCIAL PSYCHOLOGY BULLETIN LA English DT Article ID PERSONALITY-SCALES; CIRCLE; TAXONOMY; STRESS; CLASSIFICATION; PREDICTORS; DEPRESSION; CIRCUMPLEX; BEHAVIORS; DOMAIN AB The present article applies Cobb's original formulation of the social support construct and elaborates a theoretical and empirical framework employing the interpersonal circumplex model that has proved useful for elucidating concepts and measures from a variety of research traditions. In a series of studies, 1,040 undergraduate students completed self-report questionnaires indicating their likelihood of performing various support actions. A circumplex structure was obtained that was both substantively and structurally auspicious and that provided a taxonomic framework within which 12 extant social support subscales and IT personality characteristics were clarified. The Support Actions Scab Circumplex (SAS-C) assesses a broad range of social support behavior including both the potentially protective and deleterious effects of interpersonal transactions. C1 NIA, Gerontol Res Ctr, NIH, Bethesda, MD 20892 USA. RP Trobst, KK (reprint author), York Univ, Dept Psychol, 4700 Keele St, Toronto, ON M3J 1P3, Canada. NR 74 TC 47 Z9 49 U1 3 U2 10 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 0146-1672 J9 PERS SOC PSYCHOL B JI Pers. Soc. Psychol. Bull. PD AUG PY 2000 VL 26 IS 8 BP 971 EP 986 DI 10.1177/01461672002610007 PG 16 WC Psychology, Social SC Psychology GA 416NR UT WOS:000167787500007 ER PT J AU Wang, Y Chiou, AL Jeng, CH Yang, ST Lin, JC AF Wang, Y Chiou, AL Jeng, CH Yang, ST Lin, JC TI Ethanol potentiates dopamine release during acute hypoxia in rat striatum SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE ethanol; dopamine; hypoxia; striatum; alcohol; voltammetry ID METHYL-D-ASPARTATE; GLUTAMATERGIC NEUROTRANSMISSION; ASCORBATE RELEASE; BRAIN ASCORBATE; AMINO-ACIDS; IN-VITRO; ISCHEMIA; NEURONS; DAMAGE; NMDA AB We, and others, have previously demonstrated that N-methyl-D-aspartate (NMDA) receptor is involved in hypoxia or ischemia-mediated responses. We found that the NMDA antagonist ketamine attenuates cortical nitric oxide release during cerebroischemia. It has been reported that ethanol (EtOH) antagonizes NMDA-induced responses in various systems. In the present study, the interaction of EtOH and KCl-evoked striatal dopamine release in vivo during acute hypoxia was examined. High-speed chronoamperometric recording techniques, using Nafion-coated carbon fiber electrodes, were used to evaluate extracellular dopamine (DA) concentration in the striatum of urethane-anesthetized Sprague-Dawley rats. KCI was directly applied to the striatum to evoke release of DA. These anesthetized animals were paralyzed with d-tubocurarine and connected to a respirator to allow controlled respiration. Systemic concentrations of oxygen were altered by changing the rate of the respirator. We previously reported that lowering the respiratory rates from 90 to 20 times/min for 5 min decreased arterial PO2 and facilitated KCl-induced DA release in the striatum. In this study, we found that application of NMDA antagonist MK801 attenuates hypoxic DA release, suggesting that NMDA receptor is involved in this hypoxic reaction. In contrast, EtOH dose dependently enhanced KCl-evoked DA release during hypoxia. To further examine the interactions of excitatory amino acid and EtOH on DA release, glutamate was locally applied to the striatum. Glutamate-induced DA release was not affected by the systemic application of EtOH. Taken together, these data suggest that EtOH enhances DA release in vivo during short-term hypoxia, possibly through mechanisms other than excitatory amino acid pathways. (C) 2000 Elsevier Science Inc. C1 NIDA, Baltimore, MD 21042 USA. Natl Def Med Ctr, Dept Pharmacol, Taipei 100, Taiwan. Natl Def Med Ctr, Dept Neurol, Taipei 100, Taiwan. RP Wang, Y (reprint author), NIDA, 5500 Nathan Shock Dr, Baltimore, MD 21042 USA. NR 37 TC 2 Z9 2 U1 1 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 2000 VL 66 IS 4 BP 679 EP 685 DI 10.1016/S0091-3057(00)00224-0 PG 7 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 354CZ UT WOS:000089313400002 PM 10973503 ER PT J AU Hutchinson, AC Simpson, GR Randall, JF Zhang, XY Calderon, SN Rice, KC Riley, AL AF Hutchinson, AC Simpson, GR Randall, JF Zhang, XY Calderon, SN Rice, KC Riley, AL TI Assessment of SNC 80 and naltrindole within a conditioned taste aversion design SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE SNC 80; naltrindole; delta receptor; conditioned taste aversion; rat ID DELTA-OPIOID RECEPTOR; MESSAGE-ADDRESS CONCEPT; PLACE-PREFERENCE; MOTIVATIONAL PROPERTIES; ENDOGENOUS OPIOIDS; PSYCHOACTIVE DRUGS; POISON AVOIDANCE; MORPHINE; RAT; AGONIST AB Although compounds with relative selectivity for the mu and kappa opiate receptors subtypes have been reported to condition taste aversions, it is not known whether systemically administered delta compounds have the ability to produce aversions. To that end, female Long-Evans rats were adapted to water deprivation and were given pairings of a novel saccharin solution and various doses of the selective delta agonist SNC 80 (0.32-10.0 mg/kg; Experiment 1) or the selective delta antagonist naltrindole (1.0-18.0 mg/kg; Experiment 2). For comparison, the relatively selective mu agonist morphine (Experiment 1) and mu antagonist naloxone (Experiment 2) were assessed under identical conditions. Both SNC 80 (Experiment 1) and naltrindole (Experiment 2) were effective as unconditioned stimuli within this design, inducing dose-dependent taste aversions with repeated conditioning trials. Although at no dose did animals injected with SNC 80 differ from those injected with morphine, aversions induced by SNC 80 were acquired at a faster rate than those induced by morphine. Subjects injected with naloxone drank significantly less than those injected with naltrindole at the 10 mg/kg dose, and aversions induced by naloxone at 5.6 and 10 mg/kg were acquired at a faster rate than those induced by naltrindole, Although the basis for opioid agonist- and antagonist-induced taste aversions is not known, the differences between aversions induced by SNC 80 and naltrindole and those induced by morphine and naloxone, respectively, may be a function of their relative selectivity for specific opiate receptor subtypes. (C) 2000 Elsevier Science Inc. C1 American Univ, Dept Psychol, Psychopharmacol Lab, Washington, DC 20016 USA. NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Hutchinson, AC (reprint author), American Univ, Dept Psychol, Psychopharmacol Lab, Washington, DC 20016 USA. RI Marion-Poll, Frederic/D-8882-2011 OI Marion-Poll, Frederic/0000-0001-6824-0180 NR 88 TC 12 Z9 12 U1 0 U2 0 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 2000 VL 66 IS 4 BP 779 EP 787 DI 10.1016/S0091-3057(00)00278-1 PG 9 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 354CZ UT WOS:000089313400015 PM 10973516 ER PT J AU Stevenson, GW Canadas, F Zhang, XY Rice, KC Riley, AL AF Stevenson, GW Canadas, F Zhang, XY Rice, KC Riley, AL TI Morphine discriminative control is mediated by the mu opioid receptor: Assessment of delta opioid substitution and antagonism SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE morphine; SNC80; naltrindole; conditioned taste aversion; rat ID NARCOTIC ANALGESIC DRUGS; TASTE-AVERSION PROCEDURE; NALORPHINE-LIKE DRUGS; CHRONIC SPINAL DOG; STIMULUS PROPERTIES; AGONIST; RAT; CHOLECYSTOKININ; PEPTIDES; METHADONE AB Morphine is an effective training drug in drug discrimination procedures. In subsequent generalization tests in which other opioids are administered, mu opioid agonists selectively substitute for the training drug. Given the relative selectivity of morphine for the mu receptor, such substitution patterns suggest that the mu opioid receptor is mediating the discriminative control of this compound. The present study assessed this selective mediation by examining the ability of the delta opioid agonist SNC80 to substitute for (and the delta opioid antagonist naltrindole to antagonize) morphine stimulus effects in rats trained to discriminate morphine from its vehicle in the conditioned taste aversion baseline of drug discrimination learning. Although morphine and methadone produced dose-related substitution for morphine (10 mg/kg), there was no evidence of substitution for morphine by SNC80 at any dose tested. Further, although naloxone (3.2 mg/kg) completely blocked the discriminative effects of morphine, naltrindole (3.2-10 mg/kg) did not significantly affect the morphine stimulus. These data suggest that the discriminative control established to morphine is mediated by its activity at the mu, but not the delta, receptor. (C) 2000 Elsevier Science Inc. C1 American Univ, Dept Psychol, Psychopharmacol Lab, Washington, DC 20016 USA. NIDDKD, Med Chem Lab, Bethesda, MD 20892 USA. RP Stevenson, GW (reprint author), American Univ, Dept Psychol, Psychopharmacol Lab, Washington, DC 20016 USA. NR 51 TC 11 Z9 11 U1 0 U2 1 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 2000 VL 66 IS 4 BP 851 EP 856 DI 10.1016/S0091-3057(00)00280-X PG 6 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 354CZ UT WOS:000089313400024 PM 10973525 ER PT J AU Schey, KL Patat, S Chignell, CF Datillo, M Wang, RH Roberts, JE AF Schey, KL Patat, S Chignell, CF Datillo, M Wang, RH Roberts, JE TI Photooxidation of lens alpha-crystallin by hypericin (active ingredient in St John's Wort) SO PHOTOCHEMISTRY AND PHOTOBIOLOGY LA English DT Article ID SEASONAL AFFECTIVE-DISORDER; APOPTOSIS; EXTRACT; GROWTH AB Hypericin is the active ingredient in the over the counter antidepressant medication St. John's Wort. Hypericin produces singlet oxygen and other excited state intermediates that indicate it should be a very efficient phototoxic agent in the eye. Furthermore it absorbs in the UV and visible range, which means it can potentially damage both the lens and the retina. Lens a-crystallin, isolated from calf lenses, was irradiated in the presence of hypericin (5 x 10(-5) M, 10 mM ammonium bicarbonate, pH 7.0) and in the presence and absence of light (>300 nm, 24 mW/cm(2)). Hypericin-induced photosensitized photopolymerization as assessed by sodium dodecylsulfate-polyacrylamide gel electrophoresis, Further analysis of the oxidative changes occurring in cw-crystallin using mass spectrometry showed specific oxidation of methionine, tryptophan and histidine residues, which increased with irradiation time. Hypericin did not damage the lens protein in the dark. Damage to cu-crystallin could undermine the integrity of the lens directly by protein denaturation and indirectly by disturbing chaperone function. Therefore, in the presence of light, hypericin can induce changes in lens protein that could lead to the formation of cataracts. Appropriate precautions should be taken to protect the eye from intense sunlight while on this antidepressant medication. C1 Fordham Univ, New York, NY 10023 USA. Med Univ S Carolina, Dept Cell & Mol Pharmacol & Expt Therapeut, Charleston, SC 29425 USA. Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. RP Roberts, JE (reprint author), Fordham Univ, Lincoln Ctr Campus,Room 813,113 W 60th St, New York, NY 10023 USA. FU NEI NIH HHS [EY-10722] NR 28 TC 47 Z9 47 U1 1 U2 6 PU AMER SOC PHOTOBIOLOGY PI AUGUSTA PA BIOTECH PARK, 1021 15TH ST, SUITE 9, AUGUSTA, GA 30901-3158 USA SN 0031-8655 J9 PHOTOCHEM PHOTOBIOL JI Photochem. Photobiol. PD AUG PY 2000 VL 72 IS 2 BP 200 EP 203 DI 10.1562/0031-8655(2000)072<0200:POLCBH>2.0.CO;2 PG 4 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 344YV UT WOS:000088789400008 PM 10946573 ER PT J AU Kornyshev, AA Leikin, S AF Kornyshev, AA Leikin, S TI Electrostatic interaction between long, rigid helical macromolecules at all interaxial angles SO PHYSICAL REVIEW E LA English DT Article ID LIQUID-CRYSTALLINE PHASES; GUANOSINE 4-STRANDED HELICES; DEBYE-HUCKEL THEORY; POLYELECTROLYTE SOLUTIONS; AQUEOUS-SOLUTIONS; DNA MESOPHASES; CHARGED RODS; FREE-ENERGY; FORCES; CHIRALITY AB We derive formulas for the electrostatic interaction between two long, rigid macromolecules that may have arbitrary surface charge patterns and cross at an arbitrary interaxial angle. We calculate the dependence of the interaction energy on the interaxial angle, on the separation, and on the precise alignment of the charge pattern on one molecule with respect to the other. We focus in particular on molecules with helical charge patterns. We report an exact, explicit expression for the energy of interaction between net-neutral helices in a nonpolar medium as well as an approximate result for charged helices immersed in an electrolyte solution. The latter result becomes exact in the asymptotic Limit of large separations. Molecular chirality of helices manifests itself in a torque that tends to twist helices in a certain direction out of parallel alignment and that has a nontrivial behavior at small interaxial angles. We illustrate the theory with the calculation of the torque between layers of idealized, DNA-like double helices in cholesteric aggregates. We propose a mechanism of the observed cholesteric-to-columnar phase transition and suggest an explanation for the observed macroscopic (0.4-5 mu m) pitch of the cholesteric phase of B-DNA. C1 Univ Calif Santa Barbara, Inst Theoret Phys, Santa Barbara, CA 93106 USA. Res Ctr Julich, D-52425 Julich, Germany. NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. RP Kornyshev, AA (reprint author), Univ Calif Santa Barbara, Inst Theoret Phys, Santa Barbara, CA 93106 USA. RI Leikin, Sergey/A-5518-2008; Kornyshev, Alexei/C-3404-2008 OI Leikin, Sergey/0000-0001-7095-0739; NR 56 TC 49 Z9 50 U1 2 U2 9 PU AMERICAN PHYSICAL SOC PI COLLEGE PK PA ONE PHYSICS ELLIPSE, COLLEGE PK, MD 20740-3844 USA SN 1063-651X J9 PHYS REV E JI Phys. Rev. E PD AUG PY 2000 VL 62 IS 2 BP 2576 EP 2596 DI 10.1103/PhysRevE.62.2576 PN B PG 21 WC Physics, Fluids & Plasmas; Physics, Mathematical SC Physics GA 348AD UT WOS:000088960800024 PM 11088738 ER PT J AU Wu, XF Hammer, JA AF Wu, XF Hammer, JA TI Making sense of melanosome dynamics in mouse melanocytes SO PIGMENT CELL RESEARCH LA English DT Review DE melanocytes; melanosomes; molecular motors; myosin ID HERMANSKY-PUDLAK-SYNDROME; MYOSIN-V; COAT COLOR; MOTOR; GENE; TRANSPORT; PROTEIN; MODELS; LINE AB Molecular motors drive most if not all organelle movements in Eukaryotic cells. These proteins are thought to bind to the organelle surface and, through the action of their mechanochemical domains, to translocate the organelle: along a cytoskeletal track. In the case of the myosin family of molecular motors, the cytoskeletal track is filamentous actin, Microtubules serve as the cytoskeletal track for the kinesins and dyneins. While a considerable amount is known about the motors and tracks responsible for the hi-directional movement of pigment granules in fish and frog melanophores, relatively little is known about how melanosomes in mammalian melanocytes are transported out the cells dendritic arbor, accumulated at the ends of these dendrites, and transferred to keratinocytes, In this short review, we focus on the use of video microscopy to address these questions in mouse melanocytes, and we describe how an analysis of melanosome dynamics within wild type and dilute melanocytes shaped our thinking regarding the role of an unconventional myosin in melanosome transport and distribution. C1 NIH, Cell Biol Lab, Bethesda, MD 20892 USA. RP Wu, XF (reprint author), NIH, Cell Biol Lab, Bldg 3,Room B1-22, Bethesda, MD 20892 USA. NR 34 TC 33 Z9 35 U1 0 U2 0 PU MUNKSGAARD INT PUBL LTD 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 2000 VL 13 IS 4 BP 241 EP 247 DI 10.1034/j.1600-0749.2000.130405.x PG 7 WC Cell Biology; Dermatology SC Cell Biology; Dermatology GA 343RG UT WOS:000088714900004 PM 10952391 ER PT J AU McGraw, SA Sellers, D Stone, E Resnicow, KA Kuester, S Fridinger, F Wechsler, H AF McGraw, SA Sellers, D Stone, E Resnicow, KA Kuester, S Fridinger, F Wechsler, H TI Measuring implementation of school programs and policies to promote healthy eating and physical activity among youth SO PREVENTIVE MEDICINE LA English DT Article DE implementation evaluation; policy adoption; process evaluation; school; physical activity; diet ID CARDIOVASCULAR HEALTH; ADOLESCENT TRIAL; NUTRITION EDUCATION; MULTICENTER TRIAL; PREVENTION CURRICULA; TEACHING MODULES; DRUG-ABUSE; CATCH; INTERVENTION; CHILD AB Background. The measurement of program implementation and policy adoption is an essential evaluation component of any health intervention program. Data on program implementation are used to monitor program progress; identify elements of a program to be strengthened or eliminated; provide accountability; and help explain program effects. Method. This paper reviews approaches to measuring the implementation of school-based programs and policy to promote physical activity and healthful eating among youth. Areas examined include classroom instruction, food service, physical activity classes, and school policies. Operational definitions of implementation and methods of collecting data are described and compared. Results. Most implementation measures are focused on two dimensions: quantity (dose or completeness) and quality (fidelity), Data collection methods include the use of teacher self-report recorded through checklists, questionnaires, and interviews. Classroom observations by a trained observer are also used. Studies of policy development have used archival records and semi-structured interviews. Conclusions. Considerable variability exists across studies in how program of implementation is defined and measured. This is in part due to the need to tie measures closely to the content and format of the intervention. More work is needed to assess and compare the reliability and validity of various approaches to measuring implementation, (C) 2000 American Health Foundation and Academic Press. C1 New England Res Inst, Watertown, MA 02472 USA. Educ Dev Ctr, Newton, MA 02458 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Emory Univ, Rollins Sch Publ Hlth, Dept Behav Sci & Hlth Educ, Atlanta, GA 30322 USA. Ctr Dis Control & Prevent, Div Adolescent & Sch Hlth, Atlanta, GA 30341 USA. RP McGraw, SA (reprint author), New England Res Inst, Watertown, MA 02472 USA. RI Schmoelz, Camilie/D-1707-2012 OI Schmoelz, Camilie/0000-0003-2221-9954 NR 78 TC 47 Z9 47 U1 5 U2 13 PU ACADEMIC PRESS INC 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 2000 VL 31 IS 2 SU S BP S86 EP S97 DI 10.1006/pmed.2000.0648 PN 2 PG 12 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 344DJ UT WOS:000088742100006 ER PT J AU Hoskins, JR Singh, SK Maurizi, MR Wickner, S AF Hoskins, JR Singh, SK Maurizi, MR Wickner, S TI Protein binding and unfolding by the chaperone ClpA and degradation by the protease ClpAP SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE molecular chaperones; green fluorescent protein; protein unfolding ID ATP-DEPENDENT PROTEASES; HEAT-SHOCK PROTEIN; ESCHERICHIA-COLI; REGULATORY SUBUNITS; MOLECULAR CHAPERONE; CRYSTAL-STRUCTURE; QUALITY-CONTROL; TAGGING SYSTEM; 20S PROTEASOME; REPLICATION AB ClpA, a bacterial member of the Clp/Hsp100 chaperone family, is an ATP-dependent molecular chaperone and the regulatory component of the ATP-dependent ClpAP protease, To study the mechanism of binding and unfolding of proteins by ClpA and translocation to ClpP, we used as a model substrate a fusion protein that joined the ClpA recognition signal from RepA to green fluorescent protein (GFP). ClpAP degrades the fusion protein in vivo and in vitro. The substrate binds specifically to ClpA in a reaction requiring ATP binding but not hydrolysis. Binding alone is not sufficient to destabilize the native structure of the GFP portion of the fusion protein. Upon ATP hydrolysis the GFP fusion protein is unfolded, and the unfolded intermediate can be sequestered by ClpA if a nonhydrolyzable analog is added to displace ATP. ATP is required for release. We found that although ClpA is unable to recognize native proteins lacking recognition signals, including GFP and rhodanese, it interacts with those same proteins when they are unfolded. Unfolded GFP is held in a nonnative conformation while associated with ClpA and its release requires ATP hydrolysis. Degradation of unfolded untagged proteins by ClpAP requires ATP even though the initial ATP-dependent unfolding reaction is bypassed. These results suggest that there are two ATP-requiring steps: an initial protein unfolding step followed by translocation of the unfolded protein to ClpP or in some cases release from the complex. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Wickner, S (reprint author), NCI, Mol Biol Lab, NIH, Bldg 37,Rm 2D19,37 Convent Dr,MSC 37-4255, Bethesda, MD 20892 USA. NR 44 TC 112 Z9 113 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 AUG 1 PY 2000 VL 97 IS 16 BP 8892 EP 8897 DI 10.1073/pnas.97.16.8892 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000026 PM 10922051 ER PT J AU Singh, SK Grimaud, R Hoskins, JR Wickner, S Maurizi, MR AF Singh, SK Grimaud, R Hoskins, JR Wickner, S Maurizi, MR TI Unfolding and internalization of proteins by the ATP-dependent proteases ClpXP and ClpAP SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ESCHERICHIA-COLI; SPECIFICITY COMPONENT; REGULATORY SUBUNITS; MOLECULAR CHAPERONE; TAGGING SYSTEM; MU-TRANSPOSASE; BINDING; DEGRADATION; SUBSTRATE; MECHANISM AB ClpX and ClpA are molecular chaperones that interact with specific proteins and, together with ClpP, activate their ATP-dependent degradation. The chaperone activity is thought to convert proteins into an extended conformation that can access the sequestered active sites of ClpP. We now show that ClpX can catalyze unfolding of a green fluorescent protein fused to a ClpX recognition motif (GFP-SsrA), Unfolding of GFP-SsrA depends on ATP hydrolysis, GFP-SsrA unfolded either by ClpX or by treatment with denaturants binds to ClpX in the presence of adenosine 5'-O-(3-thiotriphosphate) and is released slowly (t(1/2) approximate to 15 min). Unlike ClpA, ClpX cannot trap unfolded proteins in stable complexes unless they also have a high-affinity binding motif. Addition of ATP or ADP accelerates release (t(1/2) approximate to 1 min), consistent with a model in which ATP hydrolysis induces a conformation of ClpX with low affinity for unfolded substrates. Proteolytically inactive complexes of ClpXP and ClpAP unfold GFP-SsrA and translocate the protein to ClpP, where it remains unfolded. Complexes of ClpXP with translocated substrate within the ClpP chamber retain the ability to unfold GFP-SsrA. Our results suggest a bipartite mode of interaction between ClpX and substrates. ClpX preferentially targets motifs exposed in specific proteins. As the protein is unfolded by ClpX, additional motifs are exposed that facilitate its retention and favor its translocation to ClpP for degradation. C1 NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Maurizi, MR (reprint author), NCI, Cell Biol Lab, NIH, 37 Convent Dr,MSC 4255,Bldg 37 Room 1B09, Bethesda, MD 20892 USA. NR 40 TC 189 Z9 191 U1 0 U2 4 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 AUG 1 PY 2000 VL 97 IS 16 BP 8898 EP 8903 DI 10.1073/pnas.97.16.8898 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000027 PM 10922052 ER PT J AU Cinquanta, M Rovescalli, AC Kozak, CA Nirenberg, M AF Cinquanta, M Rovescalli, AC Kozak, CA Nirenberg, M TI Mouse Sebox homeobox gene expression in skin, brain, oocytes, and two-cell embryos SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE transcription factors; OG-9; rat Sebox; human Sebox; epidermis ID POLYADENYLATED RNA; MESSENGER-RNAS; MATURATION; CLONING AB Sebox is a mouse paired-like homeobox gene, previously named OG-9. Sebox genomic DNA and cDNA were cloned and sequenced. In addition, rat and human Sebox genomic DNAs were cloned and sequenced, and the predicted amino acid sequences were compared. The mouse Sebox gene was mapped to chromosome 11 near the Evi 2 locus. The mouse Sebox gene is expressed in brain, skin, ovary, and liver of mice. In the brain, the Sebox gene is expressed in the cerebral cortex and CA areas of the hippocampus, pontine nuclei, choroid plexus, and the cerebellum. Northern analysis and RNase protection assays revealed low levels of Sebox RNA in 12-day mouse embryos and higher levels in 18- and 19-day embryos. In late embryos and newborn mice, Sebox expression is localized in the epidermis. In adult mice, Sebox RNA was found in maturing oocytes and in fertilized eggs; however, the abundance of Sebox RNA is decreased in the two-cell embryo, and little or none was detected in the four-cell embryo. Hence, Sebox is a maternally expressed homeobox gene. C1 NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RP Nirenberg, M (reprint author), NHLBI, Lab Biochem Genet, NIH, Bldg 36,Room 1C-06,36 Convent Dr,MSC 4036, Bethesda, MD 20892 USA. NR 23 TC 15 Z9 18 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 AUG 1 PY 2000 VL 97 IS 16 BP 8904 EP 8909 DI 10.1073/pnas.97.16.8904 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000028 PM 10922053 ER PT J AU Garcia, AD Aravind, L Koonin, EV Moss, B AF Garcia, AD Aravind, L Koonin, EV Moss, B TI Bacterial-type DNA Holliday junction resolvases in eukaryotic viruses SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID TEMPERATURE-SENSITIVE MUTANTS; PROTEIN SECONDARY STRUCTURE; COLI RUVC PROTEIN; VACCINIA VIRUS; ESCHERICHIA-COLI; LINEAR MINICHROMOSOMES; SCHIZOSACCHAROMYCES-POMBE; HOMOLOGOUS RECOMBINATION; CONCATEMER JUNCTIONS; RESOLVING ENZYME AB Homologous DNA recombination promotes genetic diversity and the maintenance of genome integrity, yet no enzymes with specificity for the Holliday junction (HJ)-a key DNA recombination intermediate-have been purified and characterized from metazoa or their viruses. Here we identify critical structural elements of RuvC, a bacterial HJ resolvase, in uncharacterized open reading frames from poxviruses and an iridovirus. The putative vaccinia virus resolvase was expressed as a recombinant protein, affinity purified, and shown to specifically bind and cleave a synthetic HJ to yield nicked duplex molecules. Mutation of either of two conserved acidic amino acids abrogated the catalytic activity of the A22R protein without affecting HJ binding. The presence of bacterial-type enzymes in metazoan viruses raises evolutionary questions. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20892 USA. RP Garcia, AD (reprint author), NIAID, Viral Dis Lab, NIH, 4 Ctr Dr, Bethesda, MD 20892 USA. NR 44 TC 76 Z9 79 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 AUG 1 PY 2000 VL 97 IS 16 BP 8926 EP 8931 DI 10.1073/pnas.150238697 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000032 PM 10890916 ER PT J AU Dace, A Zhao, L Park, KS Furuno, T Takamura, N Nakanishi, M West, BL Hanover, JA Cheng, SY AF Dace, A Zhao, L Park, KS Furuno, T Takamura, N Nakanishi, M West, BL Hanover, JA Cheng, SY TI Hormone binding induces rapid proteasome-mediated degradation of thyroid hormone receptors SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NUCLEAR RECEPTOR; GENERALIZED RESISTANCE; C-ERBA; EXPRESSION; MUTANT; PROTEIN; CELLS; LACTACYSTIN; ANTIBODIES; KINDREDS AB The thyroid hormone 3,3',5-triiodo-L-thyronine (T3) is essential for growth, differentiation, and development. Its biological activities are mediated by T3 nuclear receptors (TRs), At present, how T3 regulates TR proteins and the resulting functional consequences are still unknown. Immunofluorescence analyses of endogenous TR in the growth hormone-producing GC cells showed that the T3-induced rapid degradation of TR was specifically blocked by lactacystin, a selective inhibitor of the ubiquitin-proteasome degradation pathway. Immunoblots demonstrated that the transfected TR beta 1 was ubiquitinated and that the ubiquitination was T3 independent. Studies with a series of truncated TR beta 1 showed that the hormone-binding domain was sufficient for the T3-induced rapid degradation of TR beta 1 by the proteasome degradation pathway. T3 also induced rapid degradation of TR beta 2 and TR alpha 1. In contrast, the stability of the non-T3-binding TR alpha 2 and naturally occurring TR beta 1 mutants that do not bind T3 was not affected by T3 treatment, indicating that hormone binding to receptor was essential for the degradation of the wild type receptors. In the presence of proteasome protease inhibitors, the levels of both total and ubiquitinated TR beta 1 protein increased, yet T3-dependent transcriptional activation and the expression of the growth hormone gene were diminished, suggesting that proteasome-mediated degradation played a novel role in modulating transcriptional activation by TR. The present study reveals a role of T3 in modulating the functions of Tn by regulating its receptor level via the ubiquitin-proteasome degradation pathway. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Lab Cellular Biochem & Biol, NIH, Bethesda, MD 20892 USA. Univ Calif San Francisco, Med Ctr, San Francisco, CA 94143 USA. Nagoya City Univ, Fac Pharmaceut Sci, Nagoya, Aichi 467, Japan. RP Cheng, SY (reprint author), NCI, Mol Biol Lab, NIH, Bldg 37,Room 2D24,37 Convent Dr MSC 4255, Bethesda, MD 20892 USA. NR 28 TC 116 Z9 117 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 AUG 1 PY 2000 VL 97 IS 16 BP 8985 EP 8990 DI 10.1073/pnas.160257997 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000042 PM 10908671 ER PT J AU Zheng, RL Ghirlando, R Lee, MS Mizuuchi, K Krause, M Craigie, R AF Zheng, RL Ghirlando, R Lee, MS Mizuuchi, K Krause, M Craigie, R TI Barrier-to-autointegration factor (BAF) bridges DNA in a discrete, higher-order nucleoprotein complex SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID RETROVIRAL DNA; RNA INTERFERENCE; CELLULAR FACTOR; C-ELEGANS; PROTEINS AB Barrier-to-autointegration factor (BAF) is a highly conserved cellular protein that was identified by its activity in protecting retroviral DNA against autointegration. We show that BAF has the property of bridging double-stranded DNA in a highly ordered nucleoprotein complex. Whereas BAF protein alone is a dimer in solution, upon binding DNA, BAF forms a dodecamer with DNA bound at multiple discrete sites in the complex. The interactions between BAF and DNA are entirely nonspecific with respect to DNA sequence. The dual interaction of BAF with DNA and LAP2, a protein associated with the nuclear lamina, suggests a role for LAP2 in chromosome organization. Consistent with this idea, RNA interference experiments with Caenorhabditis elegans reveal a defect in mitosis. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Craigie, R (reprint author), NIDDKD, Mol Biol Lab, NIH, Bldg 5,Room 301,5 Ctr Dr MSC 0560, Bethesda, MD 20892 USA. RI Ghirlando, Rodolfo/A-8880-2009; OI Krause, Michael/0000-0001-6127-3940 NR 18 TC 144 Z9 152 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 AUG 1 PY 2000 VL 97 IS 16 BP 8997 EP + DI 10.1073/pnas.150240197 PG 7 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000044 PM 10908652 ER PT J AU Clore, GM AF Clore, GM TI Accurate and rapid docking of protein-protein complexes on the basis of intermolecular nuclear Overhauser enhancement data and dipolar couplings by rigid body minimization SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID LIQUID-CRYSTALLINE MEDIUM; NMR STRUCTURES; 3-DIMENSIONAL STRUCTURES; STRUCTURE REFINEMENT; TERMINAL DOMAIN; ENZYME-I; M-R; HPR; MACROMOLECULES; SPECTROSCOPY AB A simple and rapid method is presented for solving the three-dimensional structures of protein-protein complexes in solution on the basis of experimental NMR restraints that provide the requisite translational (i.e., intermolecular nuclear Overhauser enhancement, NOE, data) and orientational (i.e., backbone H-1-N-15 dipolar couplings and intermolecular NOEs) information. Providing high-resolution structures of the proteins in the unbound state are available and no significant backbone conformational changes occur upon complexation (which can readily be assessed by analysis of dipolar couplings measured on the complex), accurate and rapid docking of the two proteins can be achieved. The method, which is demonstrated for the 40-kDa complex of enzyme I and the histidine phosphocarrier protein, involves the application of rigid body minimization using a target function comprising only three terms, namely experimental NOE-derived intermolecular interproton distance and dipolar coupling restraints, and a simple intermolecular van der Waals repulsion potential. This approach promises to dramatically reduce the amount of time and effort required to solve the structures of protein-protein complexes by NMR, and to extend the capabilities of NMR to larger protein-protein complexes, possibly up to molecular masses of 100 kDa or more. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Clore, GM (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5, Bethesda, MD 20892 USA. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 22 TC 151 Z9 155 U1 1 U2 5 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 AUG 1 PY 2000 VL 97 IS 16 BP 9021 EP 9025 DI 10.1073/pnas.97.16.9021 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000048 PM 10922057 ER PT J AU Tanaka, TS Jaradat, SA Lim, MK Kargul, GJ Wang, XH Grahovac, MJ Pantano, S Sano, Y Piao, Y Nagaraja, R Doi, H Wood, WH Becker, KG Ko, MSH AF Tanaka, TS Jaradat, SA Lim, MK Kargul, GJ Wang, XH Grahovac, MJ Pantano, S Sano, Y Piao, Y Nagaraja, R Doi, H Wood, WH Becker, KG Ko, MSH TI Genome-wide expression profiling of mid-gestation placenta and embryo using a 15,000 mouse developmental cDNA microarray SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID GENE-EXPRESSION; TRANSCRIPTIONAL PROGRAM; IDENTIFICATION; TISSUE; CELLS; HYBRIDIZATION; IMPLANTATION; PATTERNS; YEAST AB cDNA microarray technology has been increasingly used to monitor global gene expression patterns in Various tissues and cell types. However, applications to mammalian development have been hampered by the lack of appropriate cDNA collections, particularly for early developmental stages. To overcome this problem, a PCR-based cDNA library construction method was used to derive 52,374 expressed sequence tags from pre- and peri-implantation embryos, embryonic day (E) 12.5 female gonad/mesonephros, and newborn ovary. From these cDNA collections, a microarray representing 15,264 unique genes (78% novel and 22% known) was assembled. In initial applications, the divergence of placental and embryonic: gene expression profiles was assessed. At stage E12,5 of development, based on triplicate experiments, 720 genes (6.5%) displayed statistically significant differences in expression between placenta and embryo. Among 289 more highly expressed in placenta, 61 placenta-specific genes encoded, for example, a novel prolactin-like protein. The number of genes highly expressed land frequently specific) for placenta has thereby been increased 5-fold over the total previously reported, illustrating the potential of the microarrays for tissue-specific gene discovery and analysis of mammalian developmental programs. C1 NIA, Genet Lab, NIH, Baltimore, MD 21224 USA. NIA, DNA Array Unit, NIH, Baltimore, MD 21224 USA. JST, ERATO, Doi Bioasymmetry Project, Mihama Ku, Chiba 2617112, Japan. RP Ko, MSH (reprint author), NIA, Genet Lab, NIH, 333 Cassell Dr,Suite 4000, Baltimore, MD 21224 USA. RI Ko, Minoru/B-7969-2009; OI Ko, Minoru/0000-0002-3530-3015; Becker, Kevin/0000-0002-6794-6656 NR 35 TC 337 Z9 350 U1 0 U2 5 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 AUG 1 PY 2000 VL 97 IS 16 BP 9127 EP 9132 DI 10.1073/pnas.97.16.9127 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000067 PM 10922068 ER PT J AU Searfoss, AW Wickner, RB AF Searfoss, AW Wickner, RB TI 3 ' poly(A) is dispensable for translation SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DOUBLE-STRANDED-RNA; SACCHAROMYCES-CEREVISIAE; MESSENGER-RNA; ANTIVIRAL SYSTEM; CAP STRUCTURE; L-A; YEAST; VIRUS; SKI2; IDENTIFICATION AB In wild-type cells, the 3' poly(A) structure is necessary for translation of mRNA and for mRNA stability, The superkiller 2 (ski2) ski3, ski6 ski7, and ski8 mutations enhance the expression of the poly(A)- mRNAs of yeast RNA viruses. Ski2p is a DEVH-box RNA helicase and Slh1p resembles Ski2p, Both repress L-A double-stranded RNA (dsRNA) virus copy number; further suggesting that their functions may overlap, We find that slh1 Delta ski2 Delta double mutants are healthy tin the absence of viruses) and show normal rates of turnover of several cellular mRNAs, The slh1 Delta ski2 Delta strains translate electroporated nonpoly(A) mRNA with the same kinetics as polyA(+) mRNA, Thus, the translation apparatus is inherently capable of efficiently using nonpoly(A) mRNA even in the presence of normal amounts of competing poly(A)(+) mRNA, but is normally prevented from doing so by the combined action of the nonessential proteins Ski2p and Slh1p. C1 NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP NIDDKD, Lab Biochem & Genet, NIH, 8 Ctr Dr,MSC 0830, Bethesda, MD 20892 USA. EM wickner@helix.nih.gov NR 29 TC 26 Z9 30 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 AUG 1 PY 2000 VL 97 IS 16 BP 9133 EP 9137 DI 10.1073/pnas.97.16.9133 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000068 PM 10922069 ER PT J AU Nagaraju, K Raben, N Loeffler, L Parker, T Rochon, PJ Lee, E Danning, C Wada, R Thompson, C Bahtiyar, G Craft, J van Huijsduijnen, RH Plotz, P AF Nagaraju, K Raben, N Loeffler, L Parker, T Rochon, PJ Lee, E Danning, C Wada, R Thompson, C Bahtiyar, G Craft, J van Huijsduijnen, RH Plotz, P TI Conditional up-regulation of MHC class I in skeletal muscle leads to self-sustaining autoimmune myositis and myositis-specific autoantibodies SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IDIOPATHIC INFLAMMATORY MYOPATHY; ANTIGEN-PRESENTING CELLS; TRANSFER-RNA SYNTHETASE; MEDIATED CYTO-TOXICITY; T-CELLS; HISTOCOMPATIBILITY MOLECULES; SYSTEMIC AUTOIMMUNITY; MICROVASCULAR CHANGES; ABERRANT EXPRESSION; MONONUCLEAR-CELLS AB In the human inflammatory myopathies (polymyositis and dermatomyositis), the early, widespread appearance of MHC class I on the surface of muscle cells and the occurrence of certain myositis-specific autoantibodies are striking features. We have used a controllable muscle-specific promoter system to up-regulate MHC class I in the skeletal muscles of young mice. These mice develop clinical, biochemical, histological, and immunological features very similar to human myositis. The disease is inflammatory, limited to skeletal muscles, self-sustaining, more severe in females, and often accompanied by autoantibodies, including, in some mice, autoantibodies to histidyl-tRNA synthetase, the most common specificity found in the spontaneous human disease, anti-Jo-1. This model suggests that an autoimmune disease may unfold in a highly specific pattern as the consequence of an apparently nonspecific event-the sustained up-regulation of MHC class I in a tissue-and that the specificity of the autoantibodies derives not from the specificity of the stimulus, but from the context, location, and probably the duration of the stimulus. This model further suggests that the presumed order of events as an autoimmune disease develops needs to be reconsidered. C1 NIAMSD, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Bethesda, MD 20892 USA. Yale Univ, Sch Med, Dept Internal Med, New Haven, CT 06520 USA. Serono Pharmaceut Res Inst, CH-1228 Geneva, Switzerland. RP Plotz, P (reprint author), NIAMSD, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. NR 47 TC 165 Z9 180 U1 0 U2 4 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 AUG 1 PY 2000 VL 97 IS 16 BP 9209 EP 9214 DI 10.1073/pnas.97.16.9209 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 341UL UT WOS:000088608000081 PM 10922072 ER PT J AU Gulnik, SV Suvorov, LI Majer, P Erickson, JW AF Gulnik, SV Suvorov, LI Majer, P Erickson, JW TI Sensitive fluorogenic substrates for plasmepsin 2 SO PROTEIN AND PEPTIDE LETTERS LA English DT Article ID PLASMODIUM-FALCIPARUM; MALARIA PARASITE; ASPARTIC PROTEINASE; SPECIFICITY; INHIBITION; PROTEASES; TARGETS AB Plasmepsin 2 is a malarial aspartic proteinase that has been implicated in initial steps of hemoglobin degradation in parasite and thus represents an attractive antimalarial target. We designed and synthesized a number of quenched fluorogenic substrates and measured the kinetic constants of their cleavage by plasmepsin-2. High sensitivity and superior kinetic properties make the substrates suitable for different application including fluorescence plate reader. C1 NCI, Struct Biochem Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NR 12 TC 3 Z9 3 U1 0 U2 0 PU BENTHAM SCIENCE PUBL LTD PI HILVERSUM PA PO BOX 1673, 1200 BR HILVERSUM, NETHERLANDS SN 0929-8665 J9 PROTEIN PEPTIDE LETT JI Protein Pept. Lett. PD AUG PY 2000 VL 7 IS 4 BP 219 EP 223 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 348JV UT WOS:000088982500003 ER PT J AU Jung, J Lee, B AF Jung, J Lee, B TI Protein structure alignment using environmental profiles SO PROTEIN ENGINEERING LA English DT Article DE comparison; clustering; protein structure alignment; SHEBA ID RELATE 2 SETS; PERIPLASMIC RECEPTORS; CRYSTAL-STRUCTURE; FAMILIES; SEQUENCE; FOLD; CLASSIFICATION; SIMILARITIES; BINDING; IDENTIFICATION AB A new protein structure alignment procedure is described. An initial alignment is made by comparing a one-dimensional list of primary, secondary and tertiary structural features (profiles) of two proteins, without explicitly considering the three-dimensional geometry of the structures. The alignment is then iteratively refined in the second step, in which new alignments are found by three-dimensional superposition of the structures based on the current alignment. This new procedure is fast enough to do all-against-all structural comparisons routinely. The procedure sometimes finds an alignment that suggests an evolutionary relationship and which is not normally obtained if only geometry is considered. All pair-wise comparisons were made among 3539 protein structural domains that represent all known protein structures. The resulting 3539 z-scores were used to cluster the proteins, The number of main clusters increased continuously as the z-cutoff was raised, but the number of multiple-member clusters showed a maximum at z-cutoff values of 5.0 and 5.5. When a z-cutoff value of 5.0 was used, the total number of main clusters was 2043, of which only 336 clusters had more than one member. C1 NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Lee, B (reprint author), NCI, Mol Biol Lab, Div Basic Sci, NIH, 37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 45 TC 80 Z9 81 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0269-2139 J9 PROTEIN ENG JI Protein Eng. PD AUG PY 2000 VL 13 IS 8 BP 535 EP 543 DI 10.1093/protein/13.8.535 PG 9 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology GA 358NV UT WOS:000089564900002 PM 10964982 ER PT J AU Jung, JS Lee, B AF Jung, JS Lee, B TI Use of residue pairs in protein sequence-sequence and sequence-structure alignments SO PROTEIN SCIENCE LA English DT Article DE fold recognition; pair-to-pair; score matrix; sequence alignment; threading ID AMINO-ACID SUBSTITUTION; FOLD RECOGNITION; CORRELATED MUTATIONS; SECONDARY STRUCTURE; MATRICES; PREDICTION; DATABASE; ACCURACY; SEARCH; INFORMATION AB Two new sets of scoring matrices are introduced: H-2 for the protein sequence comparison and T-2 for the protein sequence-structure correlation. Each element of H-2 or T-2 measures the frequency with which a pair of amino acid types in one protein, k-residues apart in the sequence, is aligned with another pair of residues, of given amino acid types (for H-2) or in given structural states (for T-2) in other structurally homologous proteins. There are four types, corresponding to the k-values of 1 to 4, for both H-2 and T-2 These matrices were set up using a large number of structurally homologous protein pairs, with little sequence homology between the pair, that were recently generated using the structure comparison program SHEBA. The two scoring matrices were incorporated into the main body of the sequence alignment program SSEARCH in the FASTA package and tested in a fold recognition setting in which a set of 107 test sequences were aligned to each of a panel of 3,539 domains that represent all known protein structures. Six procedures were tested; the straight Smith-Waterman (SW) and FASTA procedures, which used the Blosum62 single residue type substitution matrix; BLAST and PSI-BLAST procedures, which also used the Blosum62 matrix; PASH, which used Blosum62 and H-2 matrices; and PASSC, which used Blosum62, H-2, and T-2 matrices. All procedures gave similar results when the probe and target sequences had greater than 30% sequence identity. However, when the sequence identity was below 30%, a similar structure could be found for more sequences using PASSC than using any other procedure. PASH and PSI-BLAST gave the next best results. C1 NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Lee, B (reprint author), NCI, Mol Biol Lab, Div Basic Sci, NIH, Rm 4B15,Bldg 37,37 Concent Dr MSC 4255, Bethesda, MD 20892 USA. NR 45 TC 6 Z9 6 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD AUG PY 2000 VL 9 IS 8 BP 1576 EP 1588 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 347FR UT WOS:000088917400016 PM 10975579 ER PT J AU Schall, JD Hanes, DP Taylor, TL AF Schall, JD Hanes, DP Taylor, TL TI Neural control of behavior: countermanding eye movements SO PSYCHOLOGICAL RESEARCH-PSYCHOLOGISCHE FORSCHUNG LA English DT Article ID MONKEY SUPERIOR COLLICULUS; REACTION-TIME; NEURONAL-ACTIVITY; PREFRONTAL CORTEX; DOPAMINE NEURONS; ERROR-DETECTION; INTRACORTICAL MICROSTIMULATION; FUNCTIONAL-PROPERTIES; DELAYED-RESPONSE; FIELD NEURONS AB Understanding the self-control of action entails knowledge about how actions are initiated, how planned actions are canceled and how the consequences of actions are registered. We have investigated neural correlates of these processes using the countermanding paradigm - a task that required subjects to occasionally cancel a planned speeded response, and an analysis that provides an estimate of the time needed to cancel a planned movement. By monitoring the activity of single neurons in the frontal cortex of macaque monkeys performing this task we have distinguished signals responding to the visual stimuli, other signals that control the production of movements, and still other signals that seem to monitor behavior. C1 Vanderbilt Univ, Dept Psychol, Vanderbilt Vis Res Ctr, Nashville, TN 37240 USA. NIH, Sensorimotor Res Lab, Bethesda, MD 20892 USA. Dalhousie Univ, Dept Psychol, Halifax, NS, Canada. RP Schall, JD (reprint author), Vanderbilt Univ, Dept Psychol, Vanderbilt Vis Res Ctr, Wilson Hall,111 21st Ave S, Nashville, TN 37240 USA. FU NEI NIH HHS [R01-EY08890]; NIMH NIH HHS [F31-MH11178, R01-MH55806] NR 84 TC 25 Z9 25 U1 1 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-0727 J9 PSYCHOL RES-PSYCH FO JI Psychol. Res.-Psychol. Forsch. PD AUG PY 2000 VL 63 IS 3-4 BP 299 EP 307 DI 10.1007/s004269900008 PG 9 WC Psychology, Experimental SC Psychology GA 350DJ UT WOS:000089085200008 PM 11004883 ER PT J AU Anderson, DE AF Anderson, DE TI Ambulatory monitoring of breathing SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S21 EP S21 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500094 ER PT J AU Buchanan, TW Sollers, JJ Lovallo, WR AF Buchanan, TW Sollers, JJ Lovallo, WR TI Cortisol, emotion-modulated startle and emotional memory SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 Univ Oklahoma, Ctr Hlth Sci, Norman, OK 73019 USA. NIA, Lab Personal & Cognit, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 2 U2 2 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S30 EP S30 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500126 ER PT J AU Duncan, CC Gabbay, FH Mirsky, AF AF Duncan, CC Gabbay, FH Mirsky, AF TI Sex differences in information processing in a test of sustained attention: An event-related potential analysis SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA. NIMH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S37 EP S37 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500153 ER PT J AU Gabbay, FH Duncan, CC Mirsky, AF AF Gabbay, FH Duncan, CC Mirsky, AF TI Target P300 in a three-stimulus oddball task distinguishes amphetamine-choosing from nonchoosing subjects SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 Uniformed Serv Univ Hlth Sci, NIMH, Bethesda, MD 20814 USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S41 EP S41 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500171 ER PT J AU Gabbay, FH Duncan, CC Mirsky, AF AF Gabbay, FH Duncan, CC Mirsky, AF TI Individual differences in the target P300 elicited during the continuous performance test are related to boredom susceptibility SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 Uniformed Serv Univ Hlth Sci, NIMH, Bethesda, MD 20814 USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S41 EP S41 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500172 ER PT J AU Hagemann, D Naumann, E Thayer, JF AF Hagemann, D Naumann, E Thayer, JF TI Convergent validity of different EEG reference schemes for the measurement of anterior alpha asymmetry SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Lab Personal & Cognit, Bethesda, MD 20892 USA. Univ Maryland Baltimore Cty, Baltimore, MD 21228 USA. Univ Trier, D-54286 Trier, Germany. NR 0 TC 0 Z9 0 U1 1 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S45 EP S45 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500188 ER PT J AU Hagemann, D AF Hagemann, D TI State and trait properties of resting EEG asymmetry: Consequences for research strategies SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 1 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S18 EP S18 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500079 ER PT J AU Sollers, JJ Ahern, GL Thayer, JF AF Sollers, JJ Ahern, GL Thayer, JF TI Heart rate and heart period variability changes in the intracarotid sodium amytal test SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Lab Personal & Cognit, Bethesda, MD 20892 USA. Univ Arizona, Tucson, AZ 85721 USA. NR 0 TC 3 Z9 3 U1 0 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S93 EP S93 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500378 ER PT J AU Thayer, JF Brosschot, JF AF Thayer, JF Brosschot, JF TI Delayed cardiovascular recovery: Perseverative thinking and vagal inhibition SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. Leiden Univ, Leiden, Netherlands. NR 0 TC 0 Z9 0 U1 0 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S13 EP S13 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500055 ER PT J AU Thayer, JF AF Thayer, JF TI Neurovisceral integration: A systems approach to the relationship between neuroscience and psychophysiology SO PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD AUG PY 2000 VL 37 SU 1 BP S2 EP S2 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 364PJ UT WOS:000089900500006 ER PT J AU van Gelderen, P Grandin, C Petrella, JR Moonen, CTW AF van Gelderen, P Grandin, C Petrella, JR Moonen, CTW TI Rapid three-dimensional MR imaging: Method for tracking a bolus of contrast agent through the brain SO RADIOLOGY LA English DT Article DE brain, MR; gadolinium; magnetic resonance (MR), comparative studies; magnetic resonance (MR), contrast enhancement; magnetic resonance (MR), pulse sequences; magnetic resonance (MR), technology; magnetic resonance (MR), three-dimensional ID GRADIENT-RECALLED MRI; CEREBRAL BLOOD-FLOW; MAGNETIC-SUSCEPTIBILITY; TIME; 1.5-T AB A gradient-echo three-dimensional magnetic resonance imaging technique (principles of echo shifting train of observations, or PRESTO) is presented for use in tracking bolus of paramagnetic contrast agent through the brain. The approach combines a segmented echo-planar type of acquisition with echo shifting, which leads to echo times that are longer than the repetition time. Unlike echo-planar imaging, the method maintains image resolution despite drastic T2* changes and frequency shifts. C1 NINDS, Lab Diagnost Radiol Res, Off Director, NIH, Bethesda, MD 20892 USA. NINDS, In Vivo NMR Res Ctr, NIH, Bethesda, MD 20892 USA. Univ Victor Sagelan, Dept Magnet Resonance Biol Syst, UMR 5536, Natl Ctr Sci Res, Bordeaux, France. RP van Gelderen, P (reprint author), NINDS, Lab Diagnost Radiol Res, Off Director, NIH, Bldg 10,Rm B1D-125, Bethesda, MD 20892 USA. RI Moonen, Chrit/K-4434-2016 OI Moonen, Chrit/0000-0001-5593-3121 NR 19 TC 16 Z9 16 U1 0 U2 0 PU RADIOLOGICAL SOC NORTH AMER PI EASTON PA 20TH AND NORTHAMPTON STS, EASTON, PA 18042 USA SN 0033-8419 J9 RADIOLOGY JI Radiology PD AUG PY 2000 VL 216 IS 2 BP 603 EP 608 PG 6 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 338QM UT WOS:000088430800047 PM 10924593 ER PT J AU Dunson, DB AF Dunson, DB TI Assessing overall risk in reproductive experiments SO RISK ANALYSIS LA English DT Article DE dominant lethal assay; early loss; multiple imputation; reproductive toxicity; teratology ID DOMINANT LETHAL ASSAY; DOSE-RESPONSE MODELS; DEVELOPMENTAL TOXICITY; TERATOLOGICAL EXPERIMENTS AB Toxicologists are often interested in assessing the joint effect of an exposure on multiple reproductive endpoints, including early loss, fetal death, and malformation. Exposures that occur prior to mating or extremely early in development can adversely affect the number of implantation sites or fetuses that form within each dam and may even prevent pregnancy. A simple approach fur assessing overall adverse effects in such studies is to consider fetuses or implants that fail to develop due to exposure as missing data. The missing data can be imputed, and standard methods for the analysis of quantal response data can then be used for quantitative risk assessment or testing. In this article, a new bias-corrected imputation procedure is proposed and evaluated. The procedure is straightforward to implement in standard statistical packages and has excellent operating characteristics when used in combination with a marginal model fit with generalized estimating equations. The methods are applied to data from a reproductive toxicity study of Nitrofurazone conducted by the National Toxicology Program. C1 Natl Inst Environm Hlth Sci, Biostat Branch, Res Triangle Pk, NC 27709 USA. RP Dunson, DB (reprint author), Natl Inst Environm Hlth Sci, Biostat Branch, MD A3-03,POB 12233, Res Triangle Pk, NC 27709 USA. NR 21 TC 1 Z9 1 U1 0 U2 2 PU BLACKWELL PUBL LTD PI OXFORD PA 108 COWLEY RD, OXFORD OX4 1JF, OXON, ENGLAND SN 0272-4332 J9 RISK ANAL JI Risk Anal. PD AUG PY 2000 VL 20 IS 4 BP 429 EP 437 DI 10.1111/0272-4332.204042 PG 9 WC Public, Environmental & Occupational Health; Mathematics, Interdisciplinary Applications; Social Sciences, Mathematical Methods SC Public, Environmental & Occupational Health; Mathematics; Mathematical Methods In Social Sciences GA 361VL UT WOS:000089744900003 PM 11051068 ER PT J AU Karlson, BM Ekbom, A Wacholder, S McLaughlin, JK Hsing, AW AF Karlson, BM Ekbom, A Wacholder, S McLaughlin, JK Hsing, AW TI Cancer of the upper gastrointestinal tract among patients with pernicious anemia: A case-cohort study SO SCANDINAVIAN JOURNAL OF GASTROENTEROLOGY LA English DT Article DE esophageal cancer; gastric cancer; pancreatic cancer; pernicious anemia ID RISK FACTOR; DIABETES-MELLITUS; PANCREATIC-CANCER; GASTRIC-CANCER; STOMACH-CANCER; DISEASE AB Background: An association between pernicious anemia and stomach cancer has been established in several studies. An increased risk of pancreatic and esophageal cancers has also been reported among pernicious anemia patients. The aim of this case-cohort study was to identify additional risk factors for cancer of the esophagus, stomach, and pancreas among patients with pernicious anemia. Methods: A population-based cohort of 4586 patients with pernicious anemia was linked to the Swedish Cancer Registry to identify patients who subsequently developed cancers of the esophagus, stomach, or pancreas using a case-cohort design. A subcohort consisting of 4% of the cohort was randomly selected to serve as the comparison group. Information on medical history, smoking habits, and alcohol use was retrieved from medical charts and analyzed for cancer patients and subcohort members. Results: We could not identify any risk factors other than pernicious anemia for stomach cancer. For pancreatic and esophageal cancer, younger age at diagnosis of pernicious anemia was associated with an increased risk. A prior gastric resection, smoking and alcohol abuse were more frequent among esophageal cancer cases than in the subcohort. Conclusions: We conclude that a causal relationship between pernicious anemia and subsequent development of esophageal or pancreatic cancers still remains unproven. For esophageal cancer, confounding by smoking and alcohol use is the likely explanation of earlier reports of an association. In the case of stomach cancer, both the inflammatory process, secondary to the pernicious anemia, and pernicious anemia per se may be factors leading to malignant transformation. C1 Univ Uppsala Hosp, Dept Surg, SE-75185 Uppsala, Sweden. Karolinska Inst, Dept Med Epidemiol, Stockholm, Sweden. Harvard Univ, Sch Publ Hlth, Dept Epidemiol, Boston, MA 02115 USA. NCI, Div Canc Epidemiol & Genet, Epidemiol & Biostat Program, Bethesda, MD USA. Int Epidemiol Inst, Rockville, MD USA. RP Karlson, BM (reprint author), Univ Uppsala Hosp, Dept Surg, SE-75185 Uppsala, Sweden. NR 23 TC 25 Z9 26 U1 1 U2 5 PU TAYLOR & FRANCIS AS PI OSLO PA CORT ADELERSGT 17, PO BOX 2562, SOLLI, 0202 OSLO, NORWAY SN 0036-5521 J9 SCAND J GASTROENTERO JI Scand. J. Gastroenterol. PD AUG PY 2000 VL 35 IS 8 BP 847 EP 851 PG 5 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 348CM UT WOS:000088967200011 PM 10994624 ER PT J AU Kolstad, HA Bisanti, L Roeleveld, N Baldi, R Bonde, JP Joffe, M AF Kolstad, HA Bisanti, L Roeleveld, N Baldi, R Bonde, JP Joffe, M CA ASCLEPIOS TI Time to pregnancy among male workers of the reinforced plastics industry in Denmark, Italy and The Netherlands SO SCANDINAVIAN JOURNAL OF WORK ENVIRONMENT & HEALTH LA English DT Article DE fecundity; fertility; male; occupation; solvents; styrene exposure ID SEMEN QUALITY; STYRENE; EXPOSURES; MEN; WIVES AB Objectives The relationship between occupational styrene exposure and male fecundity was examined. Methods Among 1560 Danish, Italian, and Dutch reinforced plastics workers, 220 styrene-exposed workers and 382 unexposed referents who had fathered a child were identified. A total of 768 historical styrene measurements conducted in 1970-1996 in the study companies formed the basis for semiquantitative exposure assessment in combination with measurements of urinary styrene metabolite levels. All the subjects were interviewed about work conditions and other factors potentially related to reduced fecundity. fecundity was measured as the reported time to pregnancy (number of months a couple needed to conceive their youngest child). Results A statistically nonsignificantly reduced fecundity was observed for the styrene-exposed workers [fecundity ratio 0.79, 95% confidence interval (95% CI) 0.59-1.05]. But no consistent pattern of a detrimental effect on fecundity was found when time to pregnancy was related to worktasks indicating higher styrene exposure levels or semiquantitative or quantitative measures of styrene exposure. The workers with high exposure showed a fecundity ratio of 1.09 (95% CI 0.69-1.72). Conclusions It is unlikely that styrene exposure has a strong effect on male fecundity. C1 Aarhus Univ Hosp, Dept Occupat Med, DK-8000 Aarhus C, Denmark. Local Hlth Author, Dept Epidemiol, Milan, Italy. Univ Nijmegen, Dept Epidemiol, Nijmegen, Netherlands. Natl Canc Inst, Genoa, Italy. Univ London Imperial Coll Sci Technol & Med, Sch Med, Dept Epidemiol & Publ Hlth, London, England. RP Kolstad, HA (reprint author), Aarhus Univ Hosp, Dept Occupat Med, Norrebrogade 44, DK-8000 Aarhus C, Denmark. RI Roeleveld, Nel/B-4242-2008 OI Roeleveld, Nel/0000-0002-3390-4466 NR 22 TC 11 Z9 12 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 2000 VL 26 IS 4 BP 353 EP 358 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 351JP UT WOS:000089154300011 PM 10994802 ER PT J AU Zbar, B AF Zbar, B TI Inherited epithelial tumors of the kidney: old and new diseases SO SEMINARS IN CANCER BIOLOGY LA English DT Article ID RENAL-CELL CARCINOMA; TYROSINE KINASE RECEPTOR; SUPPRESSOR GENE; GERMLINE MUTATIONS; MET PROTOONCOGENE; TRANSLOCATION; FAMILIES; CANCER; FHIT; EXPRESSION AB This review summarizes information on inherited epithelial tumors of the kidney. Emphasis is placed on identifying clinically distinct inherited forms of renal cancer because each distinct clinical syndrome defines a different renal cancer susceptibility gene. So far, two genes that predispose to epithelial cancers of the kidney have been identified, VHL and the MET proto-oncogene. Available evidence suggests that several renal cancer genes remain to be identified. C1 NCI, Immunobiol Lab, Div Basic Sci, Frederick Canc Res Facil, Frederick, MD 21702 USA. RP Zbar, B (reprint author), NCI, Immunobiol Lab, Div Basic Sci, Frederick Canc Res Facil, Frederick, MD 21702 USA. NR 38 TC 13 Z9 13 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1044-579X J9 SEMIN CANCER BIOL JI Semin. Cancer Biol. PD AUG PY 2000 VL 10 IS 4 BP 313 EP 318 DI 10.1006/scbi.2000.0151 PG 6 WC Oncology SC Oncology GA 354BW UT WOS:000089310800006 PM 10966853 ER PT J AU Goedert, JJ AF Goedert, JJ TI The epidemiology of acquired immunodeficiency syndrome malignancies SO SEMINARS IN ONCOLOGY LA English DT Review ID SARCOMA-ASSOCIATED HERPESVIRUS; EPSTEIN-BARR-VIRUS; SQUAMOUS INTRAEPITHELIAL LESIONS; NON-HODGKINS-LYMPHOMA; HIV-INFECTED INDIVIDUALS; CENTRAL-NERVOUS-SYSTEM; TO-CHILD TRANSMISSION; KAPOSIS-SARCOMA; HOMOSEXUAL MEN; HUMAN-PAPILLOMAVIRUS C1 NCI, Viral Epidemiol Branch, Rockville, MD USA. RP Goedert, JJ (reprint author), 6120 Execut Blvd,Room 8012,MSC 7248, Rockville, MD 20852 USA. NR 80 TC 168 Z9 172 U1 0 U2 3 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 2000 VL 27 IS 4 BP 390 EP 401 PG 12 WC Oncology SC Oncology GA 350JQ UT WOS:000089097900006 PM 10950365 ER PT J AU Read, JS AF Read, JS TI Preventing mother to child transmission of HIV: the role of caesarean section SO SEXUALLY TRANSMITTED INFECTIONS LA English DT Editorial Material C1 NICHHD, Pediat Adolescent & Maternal AIDS Branch, NIH, Bethesda, MD 20892 USA. RP Read, JS (reprint author), NICHHD, Pediat Adolescent & Maternal AIDS Branch, NIH, Execut Bldg,Room 4B11F,6100 Execut Blvd MSC 7510, Bethesda, MD 20892 USA. NR 15 TC 4 Z9 4 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 1368-4973 J9 SEX TRANSM INFECT JI Sex. Transm. Infect. PD AUG PY 2000 VL 76 IS 4 BP 231 EP 232 DI 10.1136/sti.76.4.231 PG 2 WC Infectious Diseases SC Infectious Diseases GA 350JU UT WOS:000089098200001 PM 11026874 ER PT J AU Cameron, BM Allen, RC Merril, C AF Cameron, BM Allen, RC Merril, C TI A prospective study of serum pseudocholinesterase levels in patients with chronic spinal pain - A preliminary study SO SPINE LA English DT Article DE chronic spinal pain; serum cholinesterase; esterase; pseudocholinesterase; acetylcholine; cholinergic ID VESICULAR ACETYLCHOLINE TRANSPORTER; IN-VIVO; EXTRACELLULAR ACETYLCHOLINE; CEREBRAL-CORTEX; RAT CORTEX; CHOLINE-ACETYLTRANSFERASE; CEREBROSPINAL-FLUID; GENDER DIFFERENCES; BRAIN; RELEASE AB Study Design, One-dimensional polyacrylamide gel electrophoresis was used to study serum esterase enzymatic activity in three groups of patients and one group of normal volunteers. Objectives. To determine whether there is a statistically significant correlation between variations of serum pseudocholinesterase and the perception of pain in patients with chronic spinal pain. Summary of Background Data, Changes in levels of cholinesterase in the extracellular space of the brain and in the cerebral spinal fluid have been found to be associated in animal pain experimentation. Methods. Ninety-three surgical patients with chronic spinal pain, six surgical control subjects operated for conditions not associated with pain, 21 normal control volunteers, and nine disabled patients receiving monetary benefits were studied. The patients were analyzed for a period of time by rating the perception of their pain with a visual assessment score at the time venous blood was drawn. Serum samples were prepared, serum pseudocholinesterase was monitored, separated, and quantified according to Alien et al.(5) Paired sample t tests were used to statistically evaluate the data. Results. A trend of correlation was noted between preoperative serum pseudocholinesterase levels and visual assessment score: serum pseudocholinesterase levels increased as visual assessment score increased. The mean preoperative serum pseudocholinesterase level of chronic spinal pain patients (1313; SE = 26), which was I significantly higher than the mean levels of the normal control volunteers (941; SE = 24; P < 0.001) and that of surgical control subjects (1018; SE = 63; P < 0.01), decreased significantly with anesthesia (P < 0.005), The mean preoperative serum pseudocholinesterase level of the surgical controls, however, remained unchanged with anesthesia. A correlation demonstrated between visual assessment score and serum pseudocholinesterase in chronic spinal pain patients was not observed in six of nine patients receiving disability payments for more than a year. Conclusions. Measurements of quantitative alterations of serum pseudocholinesterase levels may be useful in the treatment of patients with chronic spinal pain. C1 St Lukes Episcopal Hosp, Ctr Orthopaed Res & Educ, Houston, TX 77225 USA. Leading Ion Consultants, Mt Pleasant, SC USA. NIMH, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. RP Cameron, BM (reprint author), St Lukes Episcopal Hosp, Ctr Orthopaed Res & Educ, POB 20269,MC 4-183, Houston, TX 77225 USA. NR 66 TC 3 Z9 3 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0362-2436 J9 SPINE JI SPINE PD AUG 1 PY 2000 VL 25 IS 15 BP 1917 EP 1924 DI 10.1097/00007632-200008010-00009 PG 8 WC Clinical Neurology; Orthopedics SC Neurosciences & Neurology; Orthopedics GA 341UX UT WOS:000088610000008 PM 10908934 ER PT J AU Portier, CJ Sherman, CD Kopp-Schneider, A AF Portier, CJ Sherman, CD Kopp-Schneider, A TI Multistage, stochastic models of the cancer process: A general theory for calculating tumor incidence SO STOCHASTIC ENVIRONMENTAL RESEARCH AND RISK ASSESSMENT LA English DT Article ID CARCINOGENESIS AB The use of complicated models of the cancer process has been limited by the need to have a mathematical framework allowing for a general expansion of the theory of carcinogenesis while maintaining numerically tractable results. A system of ordinary differential equations is derived for calculating tumor incidence in multistage models of carcinogenesis. The resulting system of equations is applicable to nonhomogenous processes (i.e. ones in which the rates change with time, age, etc.). In addition, a solution is given for the case of instantaneous initiation. These formulae can easily be coupled with physiologically-based pharmacokinetic models to link biomarkers of exposure to cancer risks. An example is presented. C1 Natl Inst Environm Hlth Sci, Res Triangle Pk, NC 27709 USA. San Francisco State Univ, Dept Math, San Francisco, CA 94132 USA. German Canc Res Ctr, D-69009 Heidelberg, Germany. RP Portier, CJ (reprint author), Natl Inst Environm Hlth Sci, Res Triangle Pk, NC 27709 USA. RI Portier, Christopher/A-3160-2010 OI Portier, Christopher/0000-0002-0954-0279 NR 10 TC 10 Z9 10 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 1436-3240 J9 STOCH ENV RES RISK A JI Stoch. Environ. Res. Risk Assess. PD AUG PY 2000 VL 14 IS 3 BP 173 EP 179 DI 10.1007/PL00009780 PG 7 WC Engineering, Environmental; Engineering, Civil; Environmental Sciences; Statistics & Probability; Water Resources SC Engineering; Environmental Sciences & Ecology; Mathematics; Water Resources GA 350HC UT WOS:000089094100003 ER PT J AU Waalkes, MP Fox, DA States, JC Patierno, SR McCabe, MJ AF Waalkes, MP Fox, DA States, JC Patierno, SR McCabe, MJ TI Metals and disorders of cell accumulation: Modulation of apoptosis and cell proliferation SO TOXICOLOGICAL SCIENCES LA English DT Article DE arsenic; apoptosis; cell proliferation; chromium; lead; mercury ID DNA-POLYMERASE ARREST; LEAD-EXPOSURE; AUTOIMMUNE-DISEASE; SODIUM ARSENITE; CARCINOGENIC CHROMIUM; HUMAN FIBROBLASTS; T-CELLS; CALCIUM; MERCURY; DEATH C1 NIEHS, Natl Canc Inst, Comparat Carcinogenesis Lab, Inorgan Carcinogenesis Sect, Res Triangle Pk, NC 27709 USA. Univ Houston, Coll Optometry, Houston, TX USA. Univ Houston, Dept Biol & Biochem, Houston, TX USA. Univ Louisville, Sch Med, Dept Pharmacol & Toxicol, Louisville, KY 40292 USA. George Washington Univ, Med Ctr, Dept Pharmacol, Mol & Cellular Oncol Program, Washington, DC 20037 USA. Wayne State Univ, Inst Chem Toxicol, Detroit, MI 48201 USA. RP Waalkes, MP (reprint author), NIEHS, Natl Canc Inst, Comparat Carcinogenesis Lab, Inorgan Carcinogenesis Sect, POB 12233,111 Alexander Dr,MD F0-09, Res Triangle Pk, NC 27709 USA. RI States, J./H-4246-2011 NR 59 TC 73 Z9 76 U1 0 U2 4 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 2000 VL 56 IS 2 BP 255 EP 261 DI 10.1093/toxsci/56.2.255 PG 7 WC Toxicology SC Toxicology GA 343GP UT WOS:000088693600004 PM 10910982 ER PT J AU Collins, BJ Grizzle, TB Dunnick, JK AF Collins, BJ Grizzle, TB Dunnick, JK TI Toxicokinetics of phenolphthalein in male and female rats and mice SO TOXICOLOGICAL SCIENCES LA English DT Article DE phenolphthalein; toxicokinetics; phenolphthalein-glucuronide; AUC; iv; gavage; feed; multiple routes; rat; mouse ID METABOLISM AB Phenolphthalein (PTH), which has been used as the active ingredient in a number of prescription and over-the-counter laxative products, is a rodent chemical carcinogen in multiple organs in the NTP 2-year bioassay at doses of 291-2927 mg/kg. This paper describes the toxicokinetics and estimates the internal dose of PTH administered as a single iv or gavage dose, or ad libitum for 14 days in feed to F344 rats, B6C3F1 mice, p53 (+/-) mice, and C57BL mice at doses that bracketed those used in the bioassay. Plasma concentrations for free phenolphthalein (PTH-F) and phenolphthalein glucuronide (PTH-G) were obtained for each dose regimen. Total phenolphthalein (PTH-T) was calculated as the sum of the molar concentrations of PTH-F and PTH-G. Noncompartmental pharmacokinetic models were used to calculate the area under the curve (AUC) from 0 h to infinity (AUC,), clearance (C1), and oral bioavailability (F) for PTH-F; and were used to calculate AUC(infinity), t(1/2), and relative absorption (Q) for PTH-T. After iv administration, PTH-F rapidly declined in rats and mice; PTH-T rose rapidly to Cmax and slowly declined 6-8 h after dosing, with no sex-related differences for rats or mice. For feed studies, mean plasma concentration ((C) over bar infinity) and 24-h area under the curve (AUC(24h)) values were calculated. Results from feed studies showed no dose response in rat plasma PTH-F above similar to 50 mg/kg. Rat PTH-T AUC(24h) and (C) over bar(infinity) were linear with doses up to similar to 650 mg/kg. In B6C3F1 mice, PTH-F and PTH-T AUC(24h) increased nonlinearly with doses above similar to 165 mg/kg. PTH is well absorbed and readily converted to PTH-G when administered in feed to rats and mice, except at the highest bioassay doses, where PTH absorption may be saturated. C1 NIEHS, Res Triangle Pk, NC 27709 USA. Triangle Pharmaceut Inc, Durham, NC 27707 USA. RP Collins, BJ (reprint author), NIEHS, POB 12233,111 Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 18 TC 3 Z9 3 U1 0 U2 0 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 2000 VL 56 IS 2 BP 271 EP 281 DI 10.1093/toxsci/56.2.271 PG 11 WC Toxicology SC Toxicology GA 343GP UT WOS:000088693600006 PM 10910984 ER PT J AU Josyula, S Lu, LJW Salazar, JJ Nerurkar, PV Jones, AB Grady, JJ Snyderwine, EG Anderson, LM AF Josyula, S Lu, LJW Salazar, JJ Nerurkar, PV Jones, AB Grady, JJ Snyderwine, EG Anderson, LM TI DNA adducts of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in fetal tissues of patas monkeys after transplacental exposure SO TOXICOLOGY AND APPLIED PHARMACOLOGY LA English DT Article ID PROSTAGLANDIN-H SYNTHASE; MESSENGER-RNA EXPRESSION; NONHUMAN-PRIMATES; N-ACETYLTRANSFERASE; HETEROCYCLIC AMINES; MUTAGEN 2-AMINO-3-METHYLIMIDAZO<4,5-F>QUINOLINE; CYTOCHROME-P450 ENZYMES; EXTRAHEPATIC TISSUES; METABOLIC-ACTIVATION; BETA-NAPHTHOFLAVONE AB Transplacental genotoxicity of the heterocyclic amine food-derived mutagen/carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) has been investigated by P-32-postlabeling assay for IQ-DNA adducts in maternal liver, placenta, and several fetal tissues of patas monkeys, after exposure to 15, 35, or 50 mg/kg IQ near the end of gestation or to the highest dose in the first or second trimester. Dose-dependent adduct formation occurred in all tissues, with the highest levels occurring in maternal liver. Adduct amounts were similar among fetal tissues and placenta, except for lower levels in fetal brain and slightly more adducts in fetal liver. Adducts in placenta, fetal liver, lung, kidney, skin, and adrenal gland, but not in maternal liver or fetal brain, increased! significantly as gestation progressed. Pretreatment with phenobarbital, which induces CYP enzymes that detoxify IQ, decreased adducts in maternal liver and possibly placenta, but not in fetal tissues. The CYP inducer P-naphthoflavone caused a significant increase in IQ-DNA adducts in fetal lungs. Regression analysis suggested that IQ activation in maternal and fetal liver and possibly placenta contributed to adduct formation in fetal tissues; adducts in placenta and/or fetal liver were strong predictors for those in most fetal tissues. The results indicate that exposure of pregnant primates to IQ results in DNA adduct formation in most fetal tissues, especially late in gestation; that upregulation of maternal detoxification does not provide fetal protection; and that adducts in placenta indicate adduct levels in fetal tissues. (C) 2000 Academic Press. C1 NCI, Frederick Canc Res & Dev Ctr, Comparat Carcinogenesis Lab, Frederick, MD 21702 USA. Univ Texas, Med Branch, Dept Prevent Med & Community Hlth, Galveston, TX 77555 USA. NCI, Expt Carcinogenesis Lab, Bethesda, MD 20892 USA. RP Anderson, LM (reprint author), Bldg 538, Frederick, MD 21701 USA. NR 60 TC 3 Z9 3 U1 0 U2 0 PU ACADEMIC PRESS INC 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 AUG 1 PY 2000 VL 166 IS 3 BP 151 EP 160 DI 10.1006/taap.2000.8964 PG 10 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 345KV UT WOS:000088815900001 PM 10906279 ER PT J AU Frolov, VA Cho, MS Bronk, P Reese, TS Zimmerberg, J AF Frolov, VA Cho, MS Bronk, P Reese, TS Zimmerberg, J TI Multiple local contact sites are induced by GPI-linked influenza hemagglutinin during hemifusion and flickering pore formation SO TRAFFIC LA English DT Article DE electron microscopy; enveloped virus; exocytosis; fusion pore; membrane fusion; membrane structure; rapid-freezing; viral entry ID PHOSPHOLIPID-BILAYER MEMBRANES; CELL-FUSION; PLANAR MEMBRANE; PROTEIN; EVENTS; EXOCYTOSIS; VESICLES; PH AB Membrane fusion intermediates induced by the glycosyl-phosphatidylinositol-linked ectodomain of influenza hemagglutinin (GPI-HA) were investigated by rapid freeze, freeze-substitution, thin section electron microscopy, and with simultaneous recordings of whole-cell admittance and fluorescence. Upon triggering, the previously separated membranes developed numerous hourglass shaped points of membrane contact (similar to 10-130 nm waist) when viewed by electron microscopy. Stereo pairs showed close membrane contact at peaks of complementary protrusions, arising from each membrane. With HA, there were fewer contacts, but wide fusion pores. Physiological measurements showed fast lipid dye mixing between cells after acidification, and either fusion pore formation or the lack thereof (true hemifusion). For the earliest pores, a similar conductance distribution and frequency of flickering pores were detected for both HA and GPI-HA. For GPI-HA, lipid mixing was detected prior to, during, or after pore opening, whereas for HA, lipid mixing is seen only after pore opening. Our findings are consistent with a pathway wherein conformational changes in the ectodomain of HA pull membranes towards each other to form a contact site, then hemifusion and pore formation initiate in a small percentage of these contact sites. Finally, the transmembrane domain of HA is needed to complete membrane fusion for macromolecular content mixing. C1 NICHHD, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. NINDS, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. AN Frumkin Electrochem Inst, Moscow 117071, Russia. RP Zimmerberg, J (reprint author), NICHHD, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. OI Frolov, Vadim/0000-0002-0653-5669 NR 35 TC 48 Z9 48 U1 0 U2 2 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 1398-9219 J9 TRAFFIC JI Traffic PD AUG PY 2000 VL 1 IS 8 BP 622 EP 630 DI 10.1034/j.1600-0854.2000.010806.x PG 9 WC Cell Biology SC Cell Biology GA 346RG UT WOS:000088884400005 PM 11208150 ER PT J AU Carter, LB Procter, JL Dale, JK Straus, SE Cantilena, CC AF Carter, LB Procter, JL Dale, JK Straus, SE Cantilena, CC TI Description of serologic features in autoimmune lymphoproliferative syndrome SO TRANSFUSION LA English DT Article ID FAS GENE-MUTATIONS; APOPTOSIS; DISORDER AB BACKGROUND: Autoimmune lymphoproliferative syndrome (ALPS) is a recently recognized and rare disorder associated with inherited defects in the Fas gene or other regulators of lymphocyte apoptosis. It is characterized by massive lymphadenopathy; splenomegaly; autoimmunity including episodes of immune hemolytic anemia, thrombocytopenia, and neutropenia.(1) The serologic basis for immune cytopenias associated with ALPS has not been previously characterized. STUDY DESIGN AND METHODS: RBC, granulocyte, and platelet serologies for ALPS patients and hepatitis C patients were assessed. Medical records were reviewed for clinical, immunologic, serologic, and transfusion history. Testing included: DAT; serum screening for antibodies to RBCs, granulocytes, platelets, cardiolipin, penicillin-coated RBCs, and human leukocyte antigens; antibody identification and IgG subclass; RBC phenotype. RESULTS: In a cohort of 11 patients with apoptosis defects (eight with heterozygous Fas gene mutations); many had histories of hemolytic anemia (7), thrombocytopenia (4), and/or leukopenia (11); nine received steroid therapy, seven underwent splenectomy; five had been remotely transfused. On the basis of serologic testing even when they were clinically stable, nine had positive DATs; two had alloantibodies; 6 had IgG and/or IgM antibodies to cardiolipin; seven had platelet-directed antibodies; three had granulocyte-directed antibodies; none had HLA antibodies. CONCLUSIONS: Nearly all ALPS patients have antibodies directed against one or more hematopoietic cell lineages. Serologic testing is critical in the evaluation of these individuals and when transfusion is indicated, red cells that are matched for clinically significant C, E, and K antigens should be considered. C1 NIAID, Dept Transfus Med, Warren Grant Magnuson Clin Ctr, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Procter, JL (reprint author), NIH, CC, Dept Transfus Med, Bldg 10,Room 1C711,10 Ctr Dr MSC 1184, Bethesda, MD 20892 USA. NR 16 TC 9 Z9 9 U1 0 U2 1 PU AMER ASSOC BLOOD BANKS PI BETHESDA PA 8101 GLENBROOK RD, BETHESDA, MD 20814-2749 USA SN 0041-1132 J9 TRANSFUSION JI Transfusion PD AUG PY 2000 VL 40 IS 8 BP 943 EP 948 DI 10.1046/j.1537-2995.2000.40080943.x PG 6 WC Hematology SC Hematology GA 346GN UT WOS:000088862800011 PM 10960521 ER PT J AU Xu, H Gundry, SR Hancock, WW Zuppan, C Izutani, H Bailey, LL AF Xu, H Gundry, SR Hancock, WW Zuppan, C Izutani, H Bailey, LL TI Effects of pretransplant splenectomy and immunosuppression of humoral immunity in a pig-to-newborn goat cardiac xenograft model SO TRANSPLANTATION PROCEEDINGS LA English DT Article; Proceedings Paper CT 5th Congress of the International-Xenotransplantation-Association CY OCT 24-28, 1999 CL NAGOYA, JAPAN SP Int Xenotransplant Assoc, Pharmaceut Manufacturers Assoc Tokyo, Osaka Pharmaceut Manufacturers Assoc, Suzuken Mem Fdn, Novartis Pharma AG, Novartis Pharma KK, Fujisawa Healthcare Inc, Fujisawa GmbH, Fujisawa Pharmaceut Co Ltd, F Hoffmann La Roche Ltd, Nippon Roche KK, Biotransplant Inc, Nextran Inc ID REJECTION C1 Loma Linda Univ, Med Ctr, Dept Surg, Loma Linda, CA USA. Loma Linda Univ, Med Ctr, Dept Pathol, Loma Linda, CA USA. LeukoSite Inc, Cambridge, MA USA. RP Xu, H (reprint author), Armed Forces Radiobiol Res Inst, Navy NIH Transplantat & Autoimmun Branch, Bldg 42,8901 Wisconsin Ave, Bethesda, MD 20889 USA. NR 9 TC 1 Z9 1 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0041-1345 J9 TRANSPLANT P JI Transplant. Proc. PD AUG PY 2000 VL 32 IS 5 BP 1010 EP 1014 DI 10.1016/S0041-1345(00)01088-5 PG 5 WC Immunology; Surgery; Transplantation SC Immunology; Surgery; Transplantation GA 344CV UT WOS:000088740800093 PM 10936324 ER PT J AU Xu, H Gundry, SR Hancock, W Matsumiya, G Bailey, LL AF Xu, H Gundry, SR Hancock, W Matsumiya, G Bailey, LL TI Delayed cardiac xenograft rejection in a pig-to-baboon model treated with a tolerance-inducing regimen and donor bone marrow infusion SO TRANSPLANTATION PROCEEDINGS LA English DT Article; Proceedings Paper CT 5th Congress of the International-Xenotransplantation-Association CY OCT 24-28, 1999 CL NAGOYA, JAPAN SP Int Xenotransplant Assoc, Pharmaceut Manufacturers Assoc Tokyo, Osaka Pharmaceut Manufacturers Assoc, Suzuken Mem Fdn, Novartis Pharma AG, Novartis Pharma KK, Fujisawa Healthcare Inc, Fujisawa GmbH, Fujisawa Pharmaceut Co Ltd, F Hoffmann La Roche Ltd, Nippon Roche KK, Biotransplant Inc, Nextran Inc ID TOTAL LYMPHOID IRRADIATION; TRANSPLANTATION TOLERANCE; PRIMATES C1 Loma Linda Univ, Med Ctr, Dept Surg, Loma Linda, CA USA. RP Xu, H (reprint author), NIDDK, NAVY, Transplantat & Autoimmun Branch, AFRRI, Bldg 42,8901 Wisconsin Ave, Bethesda, MD 20889 USA. NR 8 TC 0 Z9 0 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0041-1345 J9 TRANSPLANT P JI Transplant. Proc. PD AUG PY 2000 VL 32 IS 5 BP 1084 EP 1085 DI 10.1016/S0041-1345(00)01175-1 PG 2 WC Immunology; Surgery; Transplantation SC Immunology; Surgery; Transplantation GA 344CV UT WOS:000088740800137 PM 10936368 ER PT J AU Wolf, YI Kondrashov, FA Koonin, EV AF Wolf, YI Kondrashov, FA Koonin, EV TI No footprints of primordial introns in a eukaryotic genome SO TRENDS IN GENETICS LA English DT Article ID EXON THEORY; EVOLUTION; GENES; PHASE C1 NIH, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. Simons Rock Coll, Great Barrington, MA 01230 USA. RP NIH, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. EM wolf@ncbi.nlm.nih.gov; fkondras@ncbi.nlm.nih.gov; koonin@ncbi.nlm.nih.gov RI Kondrashov, Fyodor Alexeevich/H-6331-2015 OI Kondrashov, Fyodor Alexeevich/0000-0001-8243-4694 NR 18 TC 21 Z9 21 U1 0 U2 0 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 2000 VL 16 IS 8 BP 333 EP 334 DI 10.1016/S0168-9525(00)02059-X PG 2 WC Genetics & Heredity SC Genetics & Heredity GA 340HA UT WOS:000088526000004 PM 10904260 ER PT J AU Maricq, HR Jennings, JR Valter, I Frederick, M Thompson, B Smith, EA Hill, R AF Maricq, HR Jennings, JR Valter, I Frederick, M Thompson, B Smith, EA Hill, R CA Raynaud's Treatment Study Investig TI Evaluation of treatment efficacy of Raynaud phenomenon by digital blood pressure response to cooling SO VASCULAR MEDICINE LA English DT Article DE attack frequency; digital blood pressure; nifedipine; Raynaud phenomenon; treatment trial ID GEOGRAPHIC-VARIATION; PREVALENCE; NIFEDIPINE AB Our previous studies have suggested that digital blood pressure response to cooling could provide a measure of the efficacy of treatments that are administered to patients with Raynaud phenomenon (RP), This method was used on 158 primary RP patients participating in a multicenter, randomized clinical trial that compared the efficacy of sustained-release nifedipine with temperature biofeedback in the treatment of RP. A pill placebo and electromyography served as controls. The response to local finger cooling was measured at 30 degrees, 20 degrees, 15 degrees and 10 degreesC in a temperature-controlled room under standardized conditions. The results showed that, at the 15 degreesC and 10 degreesC local cooling temperatures, the patients in the nifedipine group had a higher mean digital systolic blood pressure, a higher relative digital systolic blood pressure (RDSP), a smaller proportion of subjects with RDSP < 70% and a smaller proportion of subjects with a zero reopening pressure than the patients in the three other treatment groups. These results were statistically significant at 10C, the nifedipine group being significantly different from all:others (p < 0.05); no significant difference was found between the three other treatment groups. C1 Med Univ S Carolina, Div Rheumatol & Immunol, Dept Med, Charleston, SC 29425 USA. Univ Pittsburgh, Dept Psychiat & Psychol, Pittsburgh, PA USA. Clin Trials & Surveys Corp, Baltimore, MD USA. NHLBI, Bethesda, MD 20892 USA. RP Maricq, HR (reprint author), Med Univ S Carolina, Div Rheumatol & Immunol, Dept Med, 96 Jonathan Lucas St,Suite 912,POB 250623, Charleston, SC 29425 USA. FU NHLBI NIH HHS [N01-HC-25121] NR 13 TC 8 Z9 8 U1 1 U2 2 PU ARNOLD, HODDER HEADLINE PLC PI LONDON PA 338 EUSTON ROAD, LONDON NW1 3BH, ENGLAND SN 1358-863X J9 VASC MED JI Vasc. Med. PD AUG PY 2000 VL 5 IS 3 BP 135 EP 140 PG 6 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 374TX UT WOS:000165361400002 PM 11104295 ER PT J AU Karanjawala, ZE Kaariainen, H Ghosh, S Tannenbaum, J Martin, C Ally, D Tuomilehto, J Valle, T Collins, FS AF Karanjawala, ZE Kaariainen, H Ghosh, S Tannenbaum, J Martin, C Ally, D Tuomilehto, J Valle, T Collins, FS TI Complete maternal isodisomy of chromosome 8 in an individual with an early-onset ileal carcinoid turner SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE chromosome 8; uniparental disomy; isodisomy; carcinoid; imprinting ID UNIPARENTAL DISOMY; TRISOMY-8 MOSAICISM; TUMORS; IDENTIFICATION; MUTATIONS; MARKERS; DISEASE; CANCER; GENES AB Uniparental disomy (UPD) is a condition in which diploid individuals possess a chromosome pair from a single parent. In some instances, UPD causes an abnormal phenotype due to imprinting effects, reduction to homozygosity at recessive disease loci, or trisomy mosaicism, Here we report the first account of an individual with apparently nonmosaic complete maternal isodisomy of chromosome 8. This individual was identified during routine genotyping in a genome-wide search for type 2 diabetes susceptibility genes, although he does not have diabetes. He is of normal appearance, stature, and intelligence, but there is an unusual history of early onset ileal carcinoid. The discovery of other maternal UPD 8 cases will be necessary to define whether this condition causes a distinct phenotype, Am. J, Med, Genet, 93:207-210, 2000, Published 2000 Wiley-Liss, Inc.dagger C1 Natl Human Genome Res Inst, Posit Cloning Sect, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. Family Federat Finland, Dept Med Genet, Helsinki, Finland. Natl Publ Hlth Inst, Dept Epidemiol & Hlth Promot, Diabet & Genet Epidemiol Unit, Helsinki, Finland. RP Collins, FS (reprint author), Natl Human Genome Res Inst, Posit Cloning Sect, Genet & Mol Biol Branch, NIH, Bldg 31,Room 4B09,31 Ctr Dr MSC 2152, Bethesda, MD 20892 USA. NR 25 TC 12 Z9 12 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 31 PY 2000 VL 93 IS 3 BP 207 EP 210 DI 10.1002/1096-8628(20000731)93:3<207::AID-AJMG9>3.0.CO;2-A PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 330MM UT WOS:000087965900009 PM 10925383 ER PT J AU Ho, NC Lacbawan, F Francomano, CA Ho, V AF Ho, NC Lacbawan, F Francomano, CA Ho, V TI Severe hypodontia and oral xanthomas in Alagille syndrome SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Letter ID VERRUCIFORM XANTHOMA C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. Natl Univ Singapore, Dept Oral Pathol, Singapore 117548, Singapore. RP Ho, NC (reprint author), NHGRI, Med Genet Branch, NIH, 10 Ctr Dr, Bethesda, MD 20892 USA. NR 14 TC 4 Z9 4 U1 0 U2 0 PU WILEY-LISS PI HOBOKEN PA DIV JOHN WILEY & SONS INC, 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD JUL 31 PY 2000 VL 93 IS 3 BP 250 EP 252 DI 10.1002/1096-8628(20000731)93:3<250::AID-AJMG18>3.0.CO;2-A PG 3 WC Genetics & Heredity SC Genetics & Heredity GA 330MM UT WOS:000087965900018 PM 10925392 ER PT J AU Wojda, U Miller, JL AF Wojda, U Miller, JL TI Glycosylphosphatidylinositol-anchored proteins are not required for crosslinking-mediated endocytosis or transfection of avidin bioconjugates into biotinylated cells SO BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES LA English DT Article DE GPI protein; GPI microdomain; endocytosis; biotin; avidin; gene transfer vector ID CROSS-LINKING; PLASMA-MEMBRANE; TYROSINE KINASES; GENE DELIVERY; LIVING CELLS; CAVEOLAE; CD59; POLYETHYLENIMINE; MACROMOLECULES; MICRODOMAINS AB Even though glycosylphosphatidylinositol (GPI)-anchored proteins lack direct structural contact with the intracellular space, these ubiquitously expressed surface receptors activate signaling cascades and endocytosis when crosslinked by extracellular ligands. Such properties may be due to their association with membrane microdomains composed of glycosphingolipids, cholesterol and some signaling proteins. In this study, we hypothesize that GPI proteins may be required for crosslinking-mediated endocytosis of extracellular bioconjugates. To test this hypothesis, we first biotinylated the surface membranes of native K562 erythroleukemia cells versus K562 cells incapable of surface GPI protein expression. We then compared the entry of fluorescently labeled avidin or DNA condensed on polyethylenimine-avidin bioconjugates into the two biotinylated cell populations. Using fluorescence microscopy, nearly 100% efficiency of fluorescent avidin endocytosis was demonstrated in both cell types over a 24 h period. Surprisingly, plasmid DNA transfer was slightly more efficient among the biotinylated GPI-negative cells as measured by the expression of green fluorescence protein. Our findings that GPI proteins are not required for the endocytosis of avidin bioconjugates into biotinylated cells suggest that endocytosis associated with general membrane crosslinking may be due to overall reorganization of the membrane domains rather than GPI protein-specific interactions. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. RP Miller, JL (reprint author), NIDDKD, Biol Chem Lab, NIH, 9000 Rockville Pike,Bldg 10,Rm 9B17, Bethesda, MD 20892 USA. RI Wojda, Urszula/M-6079-2015 OI Wojda, Urszula/0000-0002-4525-2004 NR 37 TC 0 Z9 0 U1 0 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0005-2736 J9 BBA-BIOMEMBRANES JI Biochim. Biophys. Acta-Biomembr. PD JUL 31 PY 2000 VL 1467 IS 1 BP 144 EP 152 DI 10.1016/S0005-2736(00)00213-3 PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 344BR UT WOS:000088738200015 PM 10930517 ER PT J AU Moody, TW Leyton, J Zia, F Tuthill, C Badamchian, M Goldstein, AL AF Moody, TW Leyton, J Zia, F Tuthill, C Badamchian, M Goldstein, AL TI Thymosin alpha 1 is chemopreventive for lung adenoma formation in A/J mice SO CANCER LETTERS LA English DT Article DE thymosin alpha 1; lung adenomas; chemoprevention; A/J mice ID GASTRIN-RELEASING PEPTIDE; AUTOCRINE GROWTH-FACTORS; CARCINOMA CELL-LINE; MONOCLONAL-ANTIBODY; CANCER GROWTH; BOMBESIN; RECEPTORS; MOUSE; CARCINOGENESIS; TUMORIGENESIS AB The effects of thymosin (THN) alpha1 were investigated using the urethane injection carcinogenesis A/J mouse model. Lung adenomas were observed 2.5, 3, and 4 months after urethane injection (400 mg/kg i.p.) into female A/J mice. Daily administration of THN alpha1 (0.4 mg/kg, s.c.) reduced lung adenoma multiplicity significantly, by approximately 45, 40, and 17%, respectively, 2.5, 3, and 4 months after urethane injection. Animals treated with THN alpha1 had a significantly greater white cell density than control A/J mice. Endogenous THN alpha1-like peptides were detected in the mouse lung. By radioimmunoassay and by Western blot, prothymosin alpha, was detected in the mouse lung. By immunocytochemistry, THN alpha1-like peptides were detected in all lung compartments including the bronchus, adenoma, bronchioles, and alveoli. These results indicate that exogenous THN alpha1 prevents lung carcinogenesis in A/J mice. (C) 2000 Published by Elsevier Science Ireland Ltd. C1 NCI, Cell & Canc Biol Dept, Med Branch, Rockville, MD 20850 USA. SciClone Pharmaceut, San Mateo, CA 94404 USA. George Washington Univ, Med Ctr, Dept Biochem & Mol Biol, Washington, DC 20037 USA. RP Moody, TW (reprint author), NCI, Cell & Canc Biol Dept, Med Branch, 9610 Med Ctr Dr, Rockville, MD 20850 USA. NR 38 TC 13 Z9 14 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD JUL 31 PY 2000 VL 155 IS 2 BP 121 EP 127 DI 10.1016/S0304-3835(00)00405-5 PG 7 WC Oncology SC Oncology GA 396XA UT WOS:000166662200002 PM 10822126 ER PT J AU Puzianowska-Kuznicka, M Nauman, A Madej, A Tanski, Z Cheng, SY Nauman, J AF Puzianowska-Kuznicka, M Nauman, A Madej, A Tanski, Z Cheng, SY Nauman, J TI Expression of thyroid hormone receptors is disturbed in human renal clear cell carcinoma SO CANCER LETTERS LA English DT Article DE human renal clear cell carcinoma; thyroid hormone receptor; gene expression; protein expression; tumorigenesis ID RETINOIC ACID RECEPTORS; CO-REPRESSOR; X-RECEPTOR; PROLIFERATION; GENE; DIFFERENTIATION; LOCALIZATION; VITAMIN-D-3; COACTIVATOR; PROTEIN AB Human renal clear cell carcinoma (RCCC) accounts for up to 2% of human cancers. To find out if thyroid hormone (T3) and its receptors (TRs) play a role in tumorigenesis of RCCC, the expression of TRs was evaluated on mRNA and protein level. It was found that TR alpha (both alpha1 and alpha2) mRNA amount was significantly decreased in tumors while compared with healthy kidney tissue, and this decrease was deepest in G1 (well differentiated) RCCCs. In contrast, TR alpha1 protein was 1.6 x overexpressed in tumors. TR beta1 mRNA amount was overexpressed in 30% and significantly decreased in 70% of examined tumors. On the protein level, TR beta1 amount was 1.7 x lower in tumors than in healthy controls. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Polish Acad Sci, Med Res Ctr, Dept Endocrinol, Warsaw, Poland. Med Ctr Postgrad Educ, Dept Biochem, Warsaw, Poland. Univ Med Sch, Dept Med & Endocrinol, PL-02097 Warsaw, Poland. Reg Hosp, Dept Urol, Ostroleka, Poland. NCI, Gene Regulat Sect, Mol Biol Lab, Bethesda, MD 20892 USA. RP Nauman, J (reprint author), Polish Acad Sci, Med Res Ctr, Dept Endocrinol, Warsaw, Poland. NR 32 TC 43 Z9 48 U1 0 U2 3 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD JUL 31 PY 2000 VL 155 IS 2 BP 145 EP 152 DI 10.1016/S0304-3835(00)00416-X PG 8 WC Oncology SC Oncology GA 396XA UT WOS:000166662200005 PM 10822129 ER PT J AU Weickert, CS Webster, MJ Colvin, SM Herman, MM Hyde, TM Weinberger, DR Kleinman, JE AF Weickert, CS Webster, MJ Colvin, SM Herman, MM Hyde, TM Weinberger, DR Kleinman, JE TI Localization of epidermal growth factor receptors and putative neuroblasts in human subependymal zone SO JOURNAL OF COMPARATIVE NEUROLOGY LA English DT Article DE neurogenesis; precursor; PSA-NCAM; class III beta-tubulin; CNPase; Neu N; Hu; subventricular zone ID ADULT MAMMALIAN BRAIN; BETA-TUBULIN ISOTYPE; MULTIPOTENTIAL STEM-CELLS; POSTNATAL RAT FOREBRAIN; HUMAN NERVOUS-SYSTEM; SUBVENTRICULAR ZONE; OLFACTORY-BULB; NEURONAL MIGRATION; PROLIFERATING CELLS; ADHESION MOLECULE AB Studies in rodents and monkeys suggest that neuronal precursor cells continue to exist and differentiate well into adulthood in these species. These results challenge the long held assumption that neurogenesis does not occur in the postnatal human brain. We examined the rostral subependymal zone (SEZ) of postnatal human brain for expression of cell phenotypic markers that have been associated with neuronal precursors and neuroblasts in rodent brain. We found epidermal growth factor receptor (EGF-R) mRNA and protein to be expressed in infant, teen, young adult, and adult human SEZ. Some SEZ cells expressed the polysialic acid form of neural cell adhesion molecule (PSA-NCAM), characteristic of migrating neuroblasts, as well as class III P-tubulin and Hu protein, characteristic of neuroblasts and early neurons. These neuroblast-like cells were negative for glial fibrillary acidic protein (GFAP), 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase), and vimentin, suggesting that they were not differentiating as glia. Our results show that neuroblast-like cells exist in the human SEZ and support the theory that SEZ of postnatal human brain has neurogenic potential. Published 2000 Wiiey-Liss. Inc.(+). C1 NIMH, Clin Brain Disorders Branch, IRP, NIH, Bethesda, MD 20892 USA. Stanley Fdn Res Program, Bethesda, MD 20814 USA. RP Weickert, CS (reprint author), MSC 1385,Bldg 10,Rm 4N 308, Bethesda, MD 20892 USA. RI Shannon Weickert, Cynthia/G-3171-2011 NR 60 TC 92 Z9 94 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 0021-9967 J9 J COMP NEUROL JI J. Comp. Neurol. PD JUL 31 PY 2000 VL 423 IS 3 BP 359 EP 372 DI 10.1002/1096-9861(20000731)423:3<359::AID-CNE1>3.0.CO;2-0 PG 14 WC Neurosciences; Zoology SC Neurosciences & Neurology; Zoology GA 330ZJ UT WOS:000087991700001 PM 10870078 ER PT J AU Yarger, DE Patrick, CB Rapoport, SI Murphy, EJ AF Yarger, DE Patrick, CB Rapoport, SI Murphy, EJ TI A continuous fluorometric assay for phospholipase A(2) activity in brain cytosol SO JOURNAL OF NEUROSCIENCE METHODS LA English DT Article DE phospholipase A(2); brain; fluorometric assay; bromoenol lactone; mouse ID CALCIUM-INDEPENDENT PHOSPHOLIPASE-A2; ALZHEIMERS-DISEASE; BROMOENOL LACTONE; ACID; CELLS; INHIBITION; SUBSTRATE; BINDING; ALBUMIN; KINASE AB Alterations in phospholipase A(2) (PLA(2)) activity have been implicated in Alzheimer disease and other neurological disorders, although brain PLA(2) activity is currently measured using lengthy, non-continuous assays. We describe herein a rapid, continuous assay in which we measured PLA(2) activity in mouse brain cytosol (CB-57). Brains were homogenized in HEPES buffer (pH 7.5) and the cytosolic fraction was prepared by centrifugation at 25 000 x g for 20 min, followed by centrifugation of the supernatant at 100 000 x g for 60 min. Cytosolic protein content was determined using the Bradford assay. Pyrene labeled phosphatidylcholine was added to 50 mu g of cytosolic protein in Tris buffer (pH 8.0) containing fatty acid free-bovine serum albumin for a final assay volume of 2 mi. Assay temperature was maintained at 30 +/- 1 degrees C. The excitation wavelength was 345 nm and emission was measured at 377 nm. Fluorescence intensity was converted to molar concentrations using a standard curve. Under these conditions, bromoenol lactone inhibited up to 58% of the PLA(2) activity with an IC50 of 0.5 mu M. In a separate experiment, lack of appreciable alternative acylhydrolase activity was verified chromatographically. Using this method, brain PLA(2) activity can be measured in a continuous, rapid, and sensitive manner. (C) 2000 Published by Elsevier Science B.V. C1 NIA, Sect Brain Physiol & Metab, Bethesda, MD 20892 USA. Cumberland Coll, Dept Biol, Williamsburg, KY 40769 USA. RP Murphy, EJ (reprint author), NIA, Sect Brain Physiol & Metab, Bldg 10,Room 6C103, Bethesda, MD 20892 USA. NR 32 TC 8 Z9 9 U1 0 U2 1 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 31 PY 2000 VL 100 IS 1-2 BP 127 EP 133 DI 10.1016/S0165-0270(00)00244-2 PG 7 WC Biochemical Research Methods; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 349HP UT WOS:000089039900016 PM 11040375 ER PT J AU Ingram, DK Roth, GS Umegaki, H Ikari, H AF Ingram, DK Roth, GS Umegaki, H Ikari, H TI Development of an adenoviral vector for intracerebral delivery of the dopamine D-2 receptor SO MECHANISMS OF AGEING AND DEVELOPMENT LA English DT Article; Proceedings Paper CT 4th International NILS Workshop on Longevity Sciences CY DEC 02-03, 1999 CL AICHI, JAPAN SP Japan Fdn Aging & Hlth DE gene therapy; Parkinson's disease; Huntington's disease; motor performance; neurotransmitter receptor; striatum; memory; acetylcholine; mutant mice ID MEDIATED GENE-TRANSFER; COGNITIVE ENHANCEMENT; TRANSGENE EXPRESSION; PARKINSONS-DISEASE; TURNING BEHAVIOR; CYSTIC-FIBROSIS; RAT STRIATUM; BRAIN; THERAPY; INVIVO AB The age-related loss of striatal dopamine D-2 receptors (D2R) has been observed in numerous species, including rodents, monkeys, and man, and is partly responsible for impaired motor function in aged mammals. We have developed an adenoviral vector designed for intracerebral transfer of cDNA for D2R. Results of in vitro studies demonstrated that the vector produced abundant message for D2R and that the vector was membrane bound and capable of binding appropriate ligand. Results of in vivo studies provided clear evidence of D2R production when injected into the striatum of rats. The D2R produced were capable of binding appropriate ligand. In addition, evidence of functional receptors was produced by demonstrating apomorphine-induced rotational behavior in rats receiving a unilateral injection of the vector. Despite these successes, we have been unable to demonstrate improvement in the motor behavior of aged rats receiving bilateral injections of the vector. A major problem with this vector as with similar adenoviral vectors is the loss of expression beginning 3-5 days after injection to undetectable levels at 21 days. Because of the lack of motor functional effects in aged rats and the loss of expression of the vector, other strategies for development of the vector are being pursued. Regarding functional effects, we have examined the feasibility of manipulating hippocampal acetylcholine (ACh) release through D2R manipulation to improve memory performance. Using microdialysis, we have demonstrated in vivo in rats that treatment with a D2R agonist increases hippocampal ACh release while treatment with a D2R antagonist attentuates this effect as well as impairs performance in a complex maze task. In addition, a D2R null mutant mouse is being used to examine possible therapeutic effects of the vector. These mice show specific motor deficits. Recent studies using positron emission tomography have also demonstrated the feasibility of in vivo imaging of the vector. Thus, use of adenoviral vectors specific for neurotransmitter receptors can provide a highly useful research tool for examining age-related alterations in behavioral function and a possible strategy for therapeutic intervention. (C) 2000 Published by Elsevier Science Ireland Ltd. C1 NIA, Mol Physiol & Genet Sect, Cellular & Mol Biol Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. Nagoya Univ, Sch Med, Dept Geriatr, Showa Ku, Nagoya 4668550, Japan. Fukushimura Hosp, Aichi 4418124, Japan. RP Ingram, DK (reprint author), NIA, Mol Physiol & Genet Sect, Cellular & Mol Biol Lab, Gerontol Res Ctr, 5600 Nathan Shock Lane, Baltimore, MD 21224 USA. NR 41 TC 4 Z9 4 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0047-6374 J9 MECH AGEING DEV JI Mech. Ageing. Dev. PD JUL 31 PY 2000 VL 116 IS 2-3 BP 77 EP 93 DI 10.1016/S0047-6374(00)00113-5 PG 17 WC Cell Biology; Geriatrics & Gerontology SC Cell Biology; Geriatrics & Gerontology GA 357FP UT WOS:000089488100003 PM 10996008 ER PT J AU Ma, BY Lii, JH Allinger, NL AF Ma, BY Lii, JH Allinger, NL TI Molecular polarizabilities and induced dipole moments in molecular mechanics SO JOURNAL OF COMPUTATIONAL CHEMISTRY LA English DT Article DE bond polarizability; molecular polarizability; induced dipole moment; MM3; ab initio ID INTERMOLECULAR POTENTIALS; DYNAMICS; MODEL; WATER AB Molecular polarizabilities may be divided into either atomic contributions or bond contributions. The common way to estimate molecular polarizabilities is to assign atomic or bond parameters for each atom or bond type to fit experimental or quantum mechanical results. Ln this study we have taken a different approach. A general formula based on MM3 force constants and bond lengths was used to compute bond polarizabilities and molecular polarizabilities. New parameters for polarizabilities are not required. A fair agreement between experimental and computed molecular polarizabilities was obtained, with a RMS deviation of 0.82 Angstrom(3) (11.7%) and signed average error of 0.01 Angstrom(3) for a broad selection of 57 molecules studied. Two methods, the many-body interaction and the pair-interaction approaches, have been used to study induced dipole moments using the bond polarizabilities estimated from the new formula. The pair-interaction approximation, which involves much less computation than the many-body interaction approach, gives a satisfactory representation of induced dipole interaction. (C) 2000 John Wiley & Sons, Inc. C1 Univ Georgia, Dept Chem, Computat Ctr Mol Struct & Design, Chem Annex, Athens, GA 30602 USA. NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, Frederick, MD 21702 USA. RP Allinger, NL (reprint author), Univ Georgia, Dept Chem, Computat Ctr Mol Struct & Design, Chem Annex, Athens, GA 30602 USA. RI Ma, Buyong/F-9491-2011 OI Ma, Buyong/0000-0002-7383-719X NR 33 TC 57 Z9 57 U1 0 U2 14 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0192-8651 J9 J COMPUT CHEM JI J. Comput. Chem. PD JUL 30 PY 2000 VL 21 IS 10 BP 813 EP 825 DI 10.1002/1096-987X(20000730)21:10<813::AID-JCC1>3.0.CO;2-T PG 13 WC Chemistry, Multidisciplinary SC Chemistry GA 326KE UT WOS:000087734000001 ER PT J AU Grene, E Newton, DA Brown, EA Berzofsky, JA Gattoni-Celli, G Shearer, GM AF Grene, E Newton, DA Brown, EA Berzofsky, JA Gattoni-Celli, G Shearer, GM TI Semi-allogeneic cell hybrids stimulate HIV-1 envelope-specific cytotoxic T lymphocytes SO AIDS LA English DT Article DE HIV infection; cytotoxic T lymphocytes; antigen presenting cells; T helper cells ID MELANOMA-CELLS; ANTIGEN EXPRESSION; IN-VIVO; PEPTIDE; INDUCTION; IMMUNITY; INDIVIDUALS; VACCINATION; EPITOPE; DNA AB Objective: The present study was designed to determine whether the HLA allogeneic T helper response stimulated by semi-allogeneic cell lines could be used as an in vitro model of immune-based therapy to stimulate HIV-specific cytotoxic T lymphocytes. Design and methods: Semi-allogeneic cell hybrids were obtained by the fusion of peripheral blood mononuclear cells from HIV-infected patients with the allogeneic beta 2-microglobulin-deficient FO1-12 melanoma cell line. These hybrids were used as antigen presenting cells for HIV envelope peptide (env)-specific cytotoxic assays. Results: The hybrid cell lines express HLA class I and II antigens from both parental cells, as well as the CD86 costimulatory molecule. HIV-specific cytotoxic T lymphocyte activity was obtained when patients' peripheral blood mononuclear cells were costimulated with env peptides plus semi-allogeneic hybrids, in contrast with stimulation with either env or hybrid cells alone. Thus, the semi-allogeneic hybrids enhanced HIV-specific killing of target cells. Conclusions: Irradiated, semi-allogeneic cell hybrids engineered for individual AIDS patients provide efficient and simultaneous co-recognition of HLA allogeneic determinants and viral antigenic determinants presented by self-HLA molecules on the same antigen presenting cells and results in the generation of enhanced HIV-specific cytotoxic T lymphocyte activity. (C) 2000 Lippincott Williams & Wilkins. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. Med Univ S Carolina, Dept Med, Dept Radiat Oncol, Charleston, SC 29425 USA. Med Univ S Carolina, Dept Med, Div Infect Dis, Charleston, SC 29425 USA. NCI, Metab Branch, NIH, Bethesda, MD USA. RP Shearer, GM (reprint author), NCI, Expt Immunol Branch, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 30 TC 8 Z9 8 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 28 PY 2000 VL 14 IS 11 BP 1497 EP 1506 DI 10.1097/00002030-200007280-00005 PG 10 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 349AL UT WOS:000089021600004 PM 10983636 ER PT J AU Shrestha, S Strathdee, SA Brahmbhatt, H Farzadegan, H Vlahov, D Smith, MW AF Shrestha, S Strathdee, SA Brahmbhatt, H Farzadegan, H Vlahov, D Smith, MW TI Short tandem repeat methodology for genotypic identification of single-person versus multi-person use of syringes SO AIDS LA English DT Article DE syringe sharing; risk behaviour validity; forensics; self-reporting; short tandem repeat (STR) ID POLYMERASE CHAIN-REACTION; INJECTION-DRUG USERS; HIV RISK BEHAVIORS; INDIVIDUAL IDENTIFICATION; FORENSIC CASEWORK; EXCHANGE PROGRAM; HEPATITIS-B; DNA; VALIDATION; SYSTEM AB Objective: To develop laboratory methods to differentiate between single- Versus multi-person use of syringes by injection drug users. Methods: Forensic short tandem repeat (STR) genetic analysis was undertaken to cross-validate a test panel of trace blood contents from syringes representing single-versus multi-person syringe use, laboratory-simulated scenarios of needle sharing generated 34 syringe washes that were blinded for evaluation. Polymerase chain reaction was used to amplify the polymorphic STR locus D6S502 from blood trace contents in used syringes. Alleles were sized and quantified using a commercial gene sequencer. A statistical algorithm was developed to determine the number of alleles present in the amplified DNA fragments. Syringes with more than two expected alleles were considered to represent multi-person syringe use. Sensitivity, specificity and the kappa coefficient were calculated. Results: Allelic matrix-based analysis of alleles from the single STR successfully characterized single-use (n = 12) and multiple-use (n = 22) syringes with 68% sensitivity and 100% specificity upon re-analysis. The extent of agreement over and above chance (kappa = 0.6; P < 0.0001) indicated good agreement for differentiating single-versus multi-person syringe use. Conclusions: These findings suggest that improved genotypic STR analysis of syringe material could be an adjunct to methods for validating self-reported needle sharing, conducting behavioral surveillance of needle-sharing behaviors, and evaluating interventions such as needle-exchange programs. Assays based on multiple STR loci will undoubtedly improve upon the promising results obtained from laboratory simulations of needle sharing. (C) 2000 Lippincott Williams & Wilkins. C1 NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Intramural Res Support Program, Frederick, MD 21702 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Baltimore, MD USA. New York Acad Med, Ctr Urban Epidemiol Studies, New York, NY USA. RP Smith, MW (reprint author), NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Intramural Res Support Program, POB B, Frederick, MD 21702 USA. RI Strathdee, Steffanie/B-9042-2009; Smith, Michael/B-5341-2012 FU NCI NIH HHS [N01-CO-56000]; NIDA NIH HHS [DA09225, DA12568] NR 37 TC 5 Z9 5 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 28 PY 2000 VL 14 IS 11 BP 1507 EP 1513 DI 10.1097/00002030-200007280-00006 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 349AL UT WOS:000089021600005 PM 10983637 ER PT J AU Kuritzkes, DR Bassett, RL Johnson, VA Marschner, IC Eron, JJ Sommadossi, JP Acosta, EP Murphy, RL Fife, K Wood, K Bell, D Martinez, A Pettinelli, CB AF Kuritzkes, DR Bassett, RL Johnson, VA Marschner, IC Eron, JJ Sommadossi, JP Acosta, EP Murphy, RL Fife, K Wood, K Bell, D Martinez, A Pettinelli, CB CA AIDS Clin Trials Grp 370 Protocol Team TI Continued lamivudine versus delavirdine in combination with indinavir and zidovudine or stavudine in lamivudine-experienced patients: results of Adult AIDS Clinical Trials Group protocol 370 SO AIDS LA English DT Article; Proceedings Paper CT 7th Conference on Retroviruses and Opportunistic Infections CY JAN 29-FEB 03, 2000 CL SAN FRANCISCO, CALIFORNIA DE HIV-1; reverse transcriptase inhibitor; nucleoside analog; nonnucleoside reverse transcriptase inhibitor; protease inhibitor ID IMMUNODEFICIENCY-VIRUS TYPE-1; PLACEBO-CONTROLLED TRIAL; HIV-1 INFECTION; DOUBLE-BLIND; ANTIRETROVIRAL THERAPY; REVERSE-TRANSCRIPTASE; DIDANOSINE; EFFICACY; NEVIRAPINE; MONOTHERAPY AB Objective: To compare the virologic activity of continued lamivudine (3TC) versus a switch to delavirdine (DLV) when initiating protease inhibitor therapy in nucleoside-experienced patients. Design: Randomized, open-label, multi-center study. Setting: Adult AIDS clinical trials units. Patients: Protease and non-nucleoside reverse transcriptase inhibitor-naive patients who had received 3TC plus zidovudine (ZDV), stavudine (d4T), or didanosine (ddl) for at least 24 weeks. Interventions: Patients with plasma HIV-1 RNA levels > 500 copies/mi who previously received d4T + 3TC or ddl + 3TC were randomized to ZDV + 3TC + indinavir (IDV) or ZDV + DLV + IDV. Main outcome measures: Primary endpoints were the proportion of patients with plasma HIV-1 RNA levels less than or equal to 200 copies/ml at 24 weeks, and occurrence of serious adverse events. The proportion of patients with plasma HIV-1 RNA levels less than or equal to 200 copies/ml at week 48 was a secondary endpoint. Results: At week 24, 58% of subjects in the ZDV + 3TC + IDV arm and 73% in the ZDV + DLV + IDV arm had plasma HIV-1 RNA levels less than or equal to 200 copies/ml (P = 0.29). At week 48, plasma HIV-1 RNA levels were less than or equal to 200 copies/ml in 48% and 83%, respectively (P = 0.007). Rash and hyperbilirubinemia occurred more frequently in DLV arm than in the 3TC arm. Steady-state plasma IDV levels were higher among patients in the DLV arm as compared with the 3TC arm. Conclusions: Substituting DLV for 3TC when adding IDV improved virologic outcome in nucleoside-experienced patients. This result might be explained, in part, by the positive effect of DLV on IDV pharmacokinetics. (C) 2000 Lippincott Williams & Wilkins. C1 Univ Colorado, Hlth Sci Ctr, Div Infect Dis, Denver, CO 80262 USA. Harvard Univ, Sch Publ Hlth, Ctr Biostat & AIDS Res, Boston, MA 02115 USA. Univ Alabama, Sch Med, Birmingham, AL USA. Vet Affairs Med Ctr, Birmingham, AL USA. Univ N Carolina, Chapel Hill, NC USA. Northwestern Univ, Chicago, IL 60611 USA. Indiana Univ, Indianapolis, IN 46204 USA. Frontier Sci & Technol Res Fdn, Amherst, NY USA. Adult AIDS Clin Trials Grp Operat Ctr, Rockville, MD USA. NIAID, Div Aids, Bethesda, MD 20892 USA. RP Kuritzkes, DR (reprint author), Univ Colorado, Hlth Sci Ctr, Div Infect Dis, 4200 E 9th Ave,B-168, Denver, CO 80262 USA. OI Murphy, Robert/0000-0003-3936-2052 FU NIAID NIH HHS [AI-25859, AI-38855, AI-38858] NR 28 TC 15 Z9 15 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 28 PY 2000 VL 14 IS 11 BP 1553 EP 1561 DI 10.1097/00002030-200007280-00011 PG 9 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 349AL UT WOS:000089021600010 PM 10983642 ER PT J AU Lewis, RA Carr, LM Doyle, K Fainstein, V Gross, R Orengo-Nania, S Samo, TC Shigley, JW Spencer, SS Weinert, M Dunn, JP Bartlett, J Becker, R Feinberg, J Jabs, DA Johnson, DA LaSalvia, S Miller, T Neisser, LG Semba, RD Tay-Kearney, ML Tucker, P Barron, B Jarrott, C Peyman, G Swenie, D Friedman, AH Ginsburg, R Sacks, H Severin, C Teich, S Wallach, F Rescigno, R Cowan, J Horan, C Kloser, P Wanner, M Friedberg, DN Addessi, A Chachoua, A Dieterich, D Hill, J Hutt, R Ligh, J Lorenzo-Latkany, M Pei, M Powers, T Scoppe, C Weinberg, DV Jampol, LM Lyon, AT Munana, A Murphy, R Palella, F Richine, L Strugala, Z Valadez, G Holland, GN Carlson, ME Chafey, S Hardy, WD Johiro, AK MacArthur-Chang, LJ Martin, MA Moe, AA Strong, CA Tufail, A Ugalat, PS Weisz, JM Freeman, WR Arevalo-Colina, JF Clark, T Jarman, CL Meixner, L Meng, TC Spector, S Taskintuna, I Torriani, FJ O'Donnell, J Alfred, P Ballesteros, F Clay, D Coleman, R Gordon, K Gumbley, D Hoffman, J Irvine, A Jacobson, M Larson, J Macalalag, L Narahara, M Payne, M Seiff, S Wilson, S Woodring, H Davis, J Mendez, P Murray, T Simmons, T van der Horst, C Kylstra, J Wohl, D Ziman, K Pavan, PR Bergen, GA Cohen, SM Craig, JA Dehler, RL Elbert, E Fox, RW Grizzard, WS Hammer, ME Hernandez, LS Herrera, S Holt, D Kemp, S Larkin, JA Ledford, DK Lockey, RF Menosky, MM Millard, S Nadler, JP Nelson, RP Norris, D Ormerod, LD Pautler, SE Poblete, SJ Rodriguez, D Rosenbach, KP Seekins, DW Toney, JR Jabs, DA Dodge, JM Klemstine, JL Schuerholtz, TA Stevens, M Meinert, CL Amend-Libercci, D Coleson, L Collins, KL Collison, BJ Dawson, C Dodge, J Donithan, M Ewing, C Fink, N Gerczak, C Harle, J Holbrook, JT Huffman, R Isaacson, MR Gilpin, AMK Lane, M Levine, CR Martin, B Meinert, J Nowakowski, DJ Owens, RM Piantadosi, B Saah, A Smith, M Tonascia, J Van Natta, ML Davis, MD Armstrong, J Brickbauer, J Brothers, R Chop, M Hubbard, L Hurlburt, D Kastorff, L Magli, Y Neider, M Onofrey, J Stoppenbach, V Vanderhoof-Young, M Walls, M Hughes, R Kurinij, N Mowery, RL Alston, B Foulkes, M Jabs, DA Davis, MD Kurinij, N Meinert, CL Mowery, RL Jabs, DA Addessi, A Alston, B Clark, T Davis, MD Feinberg, J Freeman, W Holbrook, J Holland, GN Hubbard, L Jacobson, M Kurinij, N Lewis, RA McArthur-Chang, L Meinert, C Mowery, R Murphy, R Polsky, B Tonascia, J Jabs, DA Davis, MD Duncan, WR Feinberg, J Kessler, H Kurinij, N Lambert, AG Meinert, CL Mowery, RL Powderly, W Schnittman, S Spector, S Tonascia, J Brown, BW Conway, B Grizzle, J Nussenblatt, R Phair, JP Smith, H Whitley, R Alston, B Davis, MD Foulkes, M Jabs, DA Kurinij, N Meinert, CL Mowery, RL Tonascia, J Jabs, DA Freeman, WR Jacobson, M Murphy, R Van Natta, ML Meinert, CL Cheng, B Frost, K Lambert, AG Marco, M AF Lewis, RA Carr, LM Doyle, K Fainstein, V Gross, R Orengo-Nania, S Samo, TC Shigley, JW Spencer, SS Weinert, M Dunn, JP Bartlett, J Becker, R Feinberg, J Jabs, DA Johnson, DA LaSalvia, S Miller, T Neisser, LG Semba, RD Tay-Kearney, ML Tucker, P Barron, B Jarrott, C Peyman, G Swenie, D Friedman, AH Ginsburg, R Sacks, H Severin, C Teich, S Wallach, F Rescigno, R Cowan, J Horan, C Kloser, P Wanner, M Friedberg, DN Addessi, A Chachoua, A Dieterich, D Hill, J Hutt, R Ligh, J Lorenzo-Latkany, M Pei, M Powers, T Scoppe, C Weinberg, DV Jampol, LM Lyon, AT Munana, A Murphy, R Palella, F Richine, L Strugala, Z Valadez, G Holland, GN Carlson, ME Chafey, S Hardy, WD Johiro, AK MacArthur-Chang, LJ Martin, MA Moe, AA Strong, CA Tufail, A Ugalat, PS Weisz, JM Freeman, WR Arevalo-Colina, JF Clark, T Jarman, CL Meixner, L Meng, TC Spector, S Taskintuna, I Torriani, FJ O'Donnell, J Alfred, P Ballesteros, F Clay, D Coleman, R Gordon, K Gumbley, D Hoffman, J Irvine, A Jacobson, M Larson, J Macalalag, L Narahara, M Payne, M Seiff, S Wilson, S Woodring, H Davis, J Mendez, P Murray, T Simmons, T van der Horst, C Kylstra, J Wohl, D Ziman, K Pavan, PR Bergen, GA Cohen, SM Craig, JA Dehler, RL Elbert, E Fox, RW Grizzard, WS Hammer, ME Hernandez, LS Herrera, S Holt, D Kemp, S Larkin, JA Ledford, DK Lockey, RF Menosky, MM Millard, S Nadler, JP Nelson, RP Norris, D Ormerod, LD Pautler, SE Poblete, SJ Rodriguez, D Rosenbach, KP Seekins, DW Toney, JR Jabs, DA Dodge, JM Klemstine, JL Schuerholtz, TA Stevens, M Meinert, CL Amend-Libercci, D Coleson, L Collins, KL Collison, BJ Dawson, C Dodge, J Donithan, M Ewing, C Fink, N Gerczak, C Harle, J Holbrook, JT Huffman, R Isaacson, MR Gilpin, AMK Lane, M Levine, CR Martin, B Meinert, J Nowakowski, DJ Owens, RM Piantadosi, B Saah, A Smith, M Tonascia, J Van Natta, ML Davis, MD Armstrong, J Brickbauer, J Brothers, R Chop, M Hubbard, L Hurlburt, D Kastorff, L Magli, Y Neider, M Onofrey, J Stoppenbach, V Vanderhoof-Young, M Walls, M Hughes, R Kurinij, N Mowery, RL Alston, B Foulkes, M Jabs, DA Davis, MD Kurinij, N Meinert, CL Mowery, RL Jabs, DA Addessi, A Alston, B Clark, T Davis, MD Feinberg, J Freeman, W Holbrook, J Holland, GN Hubbard, L Jacobson, M Kurinij, N Lewis, RA McArthur-Chang, L Meinert, C Mowery, R Murphy, R Polsky, B Tonascia, J Jabs, DA Davis, MD Duncan, WR Feinberg, J Kessler, H Kurinij, N Lambert, AG Meinert, CL Mowery, RL Powderly, W Schnittman, S Spector, S Tonascia, J Brown, BW Conway, B Grizzle, J Nussenblatt, R Phair, JP Smith, H Whitley, R Alston, B Davis, MD Foulkes, M Jabs, DA Kurinij, N Meinert, CL Mowery, RL Tonascia, J Jabs, DA Freeman, WR Jacobson, M Murphy, R Van Natta, ML Meinert, CL Cheng, B Frost, K Lambert, AG Marco, M CA Studies Ocular Complications AIDS AIDS Clin Trials Grp TI Long-term follow-up of patients with AIDS treated with parenteral cidofovir for cytomegalovirus retinitis: the HPMPC Peripheral Cytomegalovirus Retinitis Trial - The Studies of Ocular Complications of AIDS Research Group in collaboration with the AIDS Clinical Trials Group SO AIDS LA English DT Article DE AIDS; cidofovir; cytomegalovirus retinitis; nephrotoxicity; uveitis ID HUMAN-IMMUNODEFICIENCY-VIRUS; COMBINATION ANTIRETROVIRAL THERAPY; RELEASE GANCICLOVIR IMPLANT; INTRAVENOUS CIDOFOVIR; MAINTENANCE THERAPY; NUCLEOTIDE ANALOG; NATURAL-HISTORY; (S)-1-<3-HYDROXY-2-(PHOSPHONYLMETHOXY)PROPYL>CYTOSINE; INFECTION; DISEASE AB Objective: To evaluate patients with cytomegalovirus (CMV) retinitis treated with intravenous cidofovir for long-term outcomes. Design: Patients with CMV retinitis enrolled in a randomized, controlled clinical trial of intravenous cidofovir as treatment for retinitis were followed for long-term outcomes, including 21 patients initially enrolled in the deferral group who received cidofovir therapy after progression of retinitis. Setting: Thirteen tertiary care clinics specializing in AIDS care and ophthalmology. Participants: Fifty-eight patients with AIDS and small peripheral CMV retinitis lesions. Interventions: Cidofovir 5 mg/kg once weekly for 2 weeks followed by low-dose maintenance cidofovir therapy (3 mg/kg) in 35 patients or high-dose maintenance (5 mg/kg) in 23 patients. Main outcome measures: Time to progression of retinitis, drug toxicities. Results: Median time to progression of retinitis was 2.5 months. Median time to discontinuation of cidofovir because of intolerance was 6.6 months, and did not differ significantly between the two maintenance doses. Median time to discontinuation of cidofovir for intolerance other than probenecid reaction was 16.3 months for patients treated with low-dose maintenance and 5.0 months for patients treated with high-dose maintenance (P = 0.021). Proteinuria of 2+ or more occurred at a rate of 1.22/person year. In patients with sufficient follow-up to determine resolution of proteinuria, 89.9% of episodes resolved, and the median time to resolution was 20 days. Rates of probenecid intolerance and of cidofovir-associated uveitis were 0.35/person-year, and 0.20/person-year, respectively. C1 Baylor Coll Med, Cullen Eye Inst, Houston, TX 77030 USA. Johns Hopkins Univ, Sch Med, Baltimore, MD USA. Louisiana State Univ, Med Ctr, New Orleans, LA USA. Mt Sinai Sch Med, New York, NY USA. New Jersey Med Sch, Newark, NJ 07103 USA. NYU, Med Ctr, New York, NY 10016 USA. Northwestern Univ, Chicago, IL 60611 USA. Univ Calif Los Angeles, Los Angeles, CA USA. Univ Calif San Diego, San Diego, CA 92103 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Univ Miami, Sch Med, Miami, FL USA. Univ N Carolina, Chapel Hill, NC USA. Univ S Florida, Tampa, FL USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Coordinating Ctr, Baltimore, MD USA. Univ Wisconsin, Fundus Photograph Reading Ctr, Madison, WI USA. McKesson Bioserv Corp, Drug Distribut Ctr, Rockville, MD USA. NEI, Bethesda, MD 20892 USA. NIAID, Bethesda, MD 20892 USA. RP Jabs, DA (reprint author), 550 N Broadway,Suite 700, Baltimore, MD 21205 USA. RI Pavan, Peter/B-6473-2013 NR 46 TC 14 Z9 15 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 28 PY 2000 VL 14 IS 11 BP 1571 EP 1581 PG 11 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 349AL UT WOS:000089021600012 ER PT J AU Jackson, JB Becker-Pergola, G Guay, LA Musoke, P Mracna, M Fowler, MG Mofenson, LM Mirochnick, M Mmiro, F Eshleman, SH AF Jackson, JB Becker-Pergola, G Guay, LA Musoke, P Mracna, M Fowler, MG Mofenson, LM Mirochnick, M Mmiro, F Eshleman, SH TI Identification of the K103N resistance mutation in Ugandan women receiving nevirapine to prevent HIV-1 vertical transmission SO AIDS LA English DT Article DE reverse transcriptase inhibitors; Africa; Uganda; HIV drug resistance; vertical transmission; clinical trials; resistance mutations ID HUMAN-IMMUNODEFICIENCY-VIRUS; TYPE-1; PHARMACOKINETICS; ZIDOVUDINE; THERAPY AB Objective: A recent trial in Uganda demonstrated that a simple, inexpensive regimen of nevirapine (NVP) prophylaxis can dramatically reduce HIV-1 vertical transmission risk. In this regimen, women receive a single dose of NVP at the onset of labor and infants receive a single dose of NVP within 72 h of birth. The objective of this study was to determine whether HIV-1 variants with NVP resistance mutations were selected in Ugandan women who received this regimen in the Phase I/II trial HIVNET 006. Methods: Reverse transcriptase (RT) sequences from plasma HIV-1 were analyzed from 15 women 6 weeks after NVP dosing. RT sequences from plasma collected prior to NVP dosing were also analyzed. Results: The K103N NVP resistance mutation was detected 6 weeks after NVP administration in three (20%) out of 15 women (95% confidence interval, 0-40%). Pre-dose samples were available from two of the three women; both pre-dose samples lacked the mutation. Other NVP resistance mutations were absent from all 15 women. Women with the K103N mutation had a longer median NVP elimination half-life, decreased median oral clearance, and increased median area under the concentration time curve than those without the mutation. An evaluable sample was obtained from one of these three women 33 months after delivery; the K103N mutation was not detected in that sample. Conclusions: This preliminary study demonstrates that HIV-1 with the RT K103N mutation can be detected in some Ugandan women following a single dose of NVP. This suggests that non-nucleoside RT inhibitor resistance may be selected in some people by single dose NVP prophylaxis. Pharmacokinetic data suggested that a more prolonged exposure to NVP after dosing may favor selection of NVP-resistant HIV-1. (C) 2000 Lippincott Williams & Wilkins. C1 Johns Hopkins Med Inst, Dept Pathol, Baltimore, MD 21205 USA. Makerere Univ, Dept Paediat, Kampala, Uganda. NICHD, Div Aids, NIH, Rockville, MD USA. Ctr Dis Control, Atlanta, GA 30333 USA. NICHD, Pediat Adolescent & Mat AIDS Branch, NIH, Rockville, MD USA. Boston Univ, Dept Pediat, Boston, MA 02215 USA. Makerere Univ, Dept Obstet & Gynaecol, Kampala, Uganda. RP Eshleman, SH (reprint author), Johns Hopkins Med Inst, Dept Pathol, Ross Bldg 646,720 Rutland Ave, Baltimore, MD 21205 USA. FU NIAID NIH HHS [N0I-AI-35173]; NICHD NIH HHS [R29 NIH-CH/HD34348] NR 13 TC 126 Z9 130 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 28 PY 2000 VL 14 IS 11 BP F111 EP F115 DI 10.1097/00002030-200007280-00001 PG 5 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 349AL UT WOS:000089021600001 PM 10983633 ER PT J AU Pirone, DM Fukuhara, S Gutkind, JS Burbelo, PD AF Pirone, DM Fukuhara, S Gutkind, JS Burbelo, PD TI SPECs, small binding proteins for Cdc42 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID WISKOTT-ALDRICH-SYNDROME; ACTIN POLYMERIZATION; PLASMA-MEMBRANE; KINASE PATHWAY; ARP2/3 COMPLEX; N-WASP; GTPASES; ACTIVATION; EFFECTOR; DOMAIN AB The Rho GTPase, Cdc42, regulates a wide variety of cellular activities including actin polymerization, focal complex assembly, and kinase signaling. We have identified a new family of very small Cdc42-binding proteins, designated SPECs (for Small Protein Effector of Cdc42), that modulates these regulatory activities. The two human members, SPEC1 and SPECS, encode proteins of 79 and 84 amino acids, respectively. Both contain a conserved N-terminal region and a centrally located CRIB (Cdc42/Rac Interactive Binding) domain. Using a yeast two-hybrid system, we found that both SPECs interact strongly with Cdc42, weakly with Rad, and not at all with RhoA. Transfection analysis revealed that SPEC1 inhibited Cdc42-induced c-Jun N-terminal kinase (JNK) activation in COS1 cells in a manner that required an intact CRIB domain. Immunofluorescence experiments in MH-3T3 fibroblasts demonstrated that both SPEC1 and SPECS showed a cortical localization and induced the formation of cell surface membrane blebs, which was not dependent on Cdc42 activity. Cotransfection experiments demonstrated that SPEC1 altered Cdc42-induced cell shape changes both in COS1 cells and in NIH-3T3 fibroblasts and that this alteration required an intact CRIB domain. These results suggest that SPECs act as novel scaffold molecules to coordinate and/or mediate Cdc42 signaling activities. C1 Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Dept Oncol, Washington, DC 20007 USA. NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Burbelo, PD (reprint author), Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Dept Oncol, Rm EG16,New Res Bldg,3970 Reservoir Rd NW, Washington, DC 20007 USA. RI Gutkind, J. Silvio/A-1053-2009; Burbelo, Peter/B-1027-2009 NR 46 TC 24 Z9 27 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 22650 EP 22656 DI 10.1074/jbc.M002832200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400010 PM 10816584 ER PT J AU Hoover, DM Mizoue, LS Handel, TM Lubkowski, J AF Hoover, DM Mizoue, LS Handel, TM Lubkowski, J TI The crystal structure of the chemokine domain of fractalkine shows a novel quaternary arrangement SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GLYCOSAMINOGLYCAN-BINDING; FUNCTIONAL EXPRESSION; CHEMOTACTIC RECEPTOR; CELL-ADHESION; IDENTIFICATION; CX(3)CR1; ACTIVATION; RANTES; FLOW; AGGREGATION AB Fractalkine, or neurotactin, is a chemokine that is present in endothelial cells from several tissues, including brain, liver, and kidney. It is the only member of the CX3C class of chemokines. Fractalkine contains a chemokine domain (CDF) attached to a membrane-spanning domain via a mucin-like stalk. However, fractalkine can also be proteolytically cleaved from its membrane-spanning domain to release a freely diffusible form. Fractalkine attracts and immobilizes leukocytes by binding to its receptor, CX(3)CR1. The x-ray crystal structure of CDF has been solved and refined to 2.0 Angstrom resolution. The CDF monomers form a dimer through an intermolecular beta-sheet, This interaction is somewhat similar to that seen in other dimeric CC chemokine crystal structures. However, the displacement of the first disulfide in CDF causes the dimer to assume a more compact quaternary structure relative to CC chemokines, which is unique to CX3C chemokines, Although fractalkine can bind to heparin in vitro, as shown by comparison of electrostatic surface plots with other chemokines and by heparin chromatography, the role of this property in vivo is not well understood. C1 NCI, Frederick Canc Res & Dev Ctr, Program Struct Biol, Macromol Crystallog Lab, Frederick, MD 21702 USA. Univ Calif Berkeley, Dept Cell & Mol Biol, Berkeley, CA 94720 USA. RP Lubkowski, J (reprint author), NCI, Frederick Canc Res & Dev Ctr, Program Struct Biol, Macromol Crystallog Lab, Frederick, MD 21702 USA. NR 43 TC 38 Z9 40 U1 0 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 23187 EP 23193 DI 10.1074/jbc.M002584200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400080 PM 10770945 ER PT J AU Higashimoto, Y Saito, S Tong, XH Hong, A Sakaguchi, K Appella, E Anderson, CW AF Higashimoto, Y Saito, S Tong, XH Hong, A Sakaguchi, K Appella, E Anderson, CW TI Human p53 is phosphorylated on serines 6 and 9 in response to DNA damage-inducing agents SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CASEIN KINASE-I; PROTEIN; GENE; EXPRESSION; PHOSPHOSERINE; MUTATIONS; ANALOGS; SIGNALS; ISOFORM; GROWTH AB To characterize the sites in human p53 that become phosphorylated in response to DNA damage, we have developed polyclonal antibodies that recognize p53 only when it is phosphorylated at specific sites. Several attempts to generate an antibody to p53 phosphorylated at Ser(6) using a phosphoserine-containing peptide as an immunogen were unsuccessful; however, phosphorylation-specific antibodies were produced by using the phosphoserine mimetic, L-2-amino-4-phosphono-4,4-di-fluorobutanoicacid (F(2)Pab), in place of phosphoserine. Fmoc-F(2)Pab was prepared by an improved synthesis and chemically incorporated using solid phase peptide synthesis, Affinity-purified antibodies elicited by immunizing rabbits with an F(2)Pab peptide coupled to keyhole limpet hemocyanin recognized a p53(1-39) peptide phosphorylated only at Ser(6) but not the unphosphorylated peptide or the same peptide phosphorylated at Ser(9), Ser(15), Ser(20), Ser(33), or Ser(37). Untreated A549 cells exhibited a background of constitutive phosphorylation at Ser(6) that increased approximately 10-fold upon exposure to either ionizing radiation or UV light. Similar results were obtained for Sers using antibodies raised against a conventional phosphopeptide. Ser(9) was phosphorylated by casein kinase 1 in vitro in a phosphoserine 6-dependent manner. Our data identify two additional DNA damage-induced phosphorylations in human p53 and show that F(2)Pab-derivatized peptides can be used to develop phosphorylation site-specific polyclonal antibodies. C1 NCI, NIH, Bethesda, MD 20892 USA. AnaSpec Inc, San Jose, CA 95131 USA. Kyushu Univ, Fac Sci, Dept Chem, Fukuoka 8128581, Japan. Brookhaven Natl Lab, Dept Biol, Upton, NY 11973 USA. RP Appella, E (reprint author), NCI, NIH, Bethesda, MD 20892 USA. RI U-ID, Kyushu/C-5291-2016 NR 32 TC 88 Z9 89 U1 0 U2 12 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 23199 EP 23203 DI 10.1074/jbc.M002674200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400082 PM 10930428 ER PT J AU Yang, TX Huang, YN Heasley, LE Berl, T Schnermann, JB Briggs, JP AF Yang, TX Huang, YN Heasley, LE Berl, T Schnermann, JB Briggs, JP TI MAPK mediation of hypertonicity-stimulated cyclooxygenase-2 expression in renal medullary collecting duct cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ACTIVATED PROTEIN-KINASE; SIGNAL-REGULATED KINASE; OSMOTIC RESPONSE ELEMENT; CANINE KIDNEY-CELLS; N-TERMINAL KINASE; EPITHELIAL-CELLS; TYROSINE PHOSPHORYLATION; MESANGIAL CELLS; MAMMALIAN-CELLS; P38 KINASE AB We have previously shown that hypertonicity stimulates cyclooxygenase-2 (COX-2) expression in cultured medullary epithelial cells. The aims of the present study were (i) to examine the role of cytoplasmic signaling through MAPK pathways in tonicity regulation of COX-2 expression in collecting duct cells and (ii) to assess the possible contribution of COX-2 to the survival of inner medullary collecting duct (IMCD) cells under hypertonic conditions. In mIMCD-K2 cells, a cell line derived from mouse IMCDs, hypertonicity induced a marked increase in COX-2 protein expression, The stimulation was reduced significantly by inhibition of MEK1 (PD-98059, 5-50 mu M and p38 (SB-203580, 5-100 mu M) and was almost abolished by the combination of the two compounds. To study the role of JNK in tonicity-stimulated COX-2 expression, IMCD-3 cell lines stably transfected with dominant-negative mutants of three JNKs (JNK-1, -2, and -3) were used. Hypertonicity-stimulated COX-2 protein expression was significantly reduced in dominant-negative JNK-2-expressing cells and was unchanged in dominant-negative JNK-1- and JNK-3-expressing cells compared with controls. The reduction of COX-2 expression was associated with greatly reduced viability of dominant-negative JNK-2-expressing cells during hypertonicity treatment, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) (2-8 mu M), an inhibitor of Src kinases, reduced the tonicity-stimulated COX-2 expression in a dose-dependent manner, whereas PP3, an inactive analog of PP2, had no effect. Inhibition of COX-2 activity by NS-398 (30-90 mu M) and SC-58236 (10-20 mu M) significantly reduced viability of mIMCD-K2 cells subjected to prolonged hypertonic treatment. We conclude that 1) all three members of the MAPK family (ERK, JNK-2, and p38) as well as Src kinases are required for tonicity-stimulated COX-2 expression in mouse collecting duct cells and that 2) COX-2 may play a role in cell survival of medullary cells under hypertonic conditions. C1 NIDDK, NIH, Bethesda, MD 20892 USA. Univ Colorado, Dept Internal Med, Denver, CO 80262 USA. RP Briggs, JP (reprint author), NIDDK, NIH, Bldg 31,Rm 9A17,31 Ctr Dr,MSC 2560, Bethesda, MD 20892 USA. RI Briggs, Josephine/B-9394-2009 OI Briggs, Josephine/0000-0003-0798-1190 FU NIDDK NIH HHS [DK-37448, DK-39255, DK-40042] NR 37 TC 92 Z9 96 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 23281 EP 23286 DI 10.1074/jbc.M910237199 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400094 PM 10930430 ER PT J AU Poltilove, RMK Jacobs, AR Haft, CR Xu, P Taylor, SI AF Poltilove, RMK Jacobs, AR Haft, CR Xu, P Taylor, SI TI Characterization of Drosophila insulin receptor substrate SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CARBOXYL-TERMINAL EXTENSION; TYROSINE KINASE; SIGNAL-TRANSDUCTION; GENE-PRODUCT; S6 KINASE; PROTEIN; GROWTH; CELL; MELANOGASTER; HOMOLOG AB Insulin receptor substrate (IRS) proteins are phosphorylated by multiple tyrosine kinases, including the insulin receptor. Phosphorylated IRS proteins bind to SH2 domain-containing proteins, thereby triggering downstream signaling pathways. The Drosophila insulin receptor (dIR) C-terminal extension contains potential binding sites for signaling molecules, suggesting that dIR might not require an IRS protein to accomplish its signaling functions. However, we obtained a cDNA encoding Drosophila IRS (dIRS), and we demonstrated expression of dIRS in a Drosophila cell line. Like mammalian IRS proteins, the N-terminal portion of dIRS contains a pleckstrin homology domain and a phosphotyrosine binding domain that binds to phosphotyrosine residues in both human and Drosophila insulin receptors. When coexpressed with dIRS in COS-7 cells, a chimeric receptor (the extracellular domain of human IR fused to the cytoplasmic domain of dIR) mediated insulin-stimulated tyrosine phosphorylation of dIRS. Mutating the juxtamembrane NPXY motif markedly reduced the ability of the receptor to phosphorylate dIRS. In contrast, the NPXY motifs in the C-terminal extension of dIR were required for stable association with dIRS. Coimmunoprecipitationexperimentsdemonstratedinsulin-dependent binding of dIRS to phosphatidylinositol 3-kinase and SHP2. However, we did not detect interactions with Grb2, SHC, or phospholipase C-gamma. Taken together with published genetic studies, these biochemical data support the hypothesis that dIRS functions directly downstream from the insulin receptor in Drosophila. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. George Washington Univ, Grad Program Genet, Washington, DC 20052 USA. RP Taylor, SI (reprint author), NIDDKD, Diabet Branch, NIH, Bldg 10,Rm 9S213, Bethesda, MD 20892 USA. NR 51 TC 18 Z9 19 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 23346 EP 23354 DI 10.1074/jbc.M003579200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400103 PM 10801879 ER PT J AU Zhang, WG Trible, RP Zhu, MH Liu, SK McGlade, J Samelson, LE AF Zhang, WG Trible, RP Zhu, MH Liu, SK McGlade, J Samelson, LE TI Association of Grb2 Gads, and phospholipase C-gamma 1 with phosphorylated LAT tyrosine residues - Effect of LAT tyrosine mutations on T cell antigen receptor-mediated signaling SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ADAPTER PROTEIN; MOLECULAR-CLONING; ACTIVATION; KINASES; PLC-GAMMA-1; SLP-76; REQUIREMENT; EXCHANGE; DOMAIN; ZAP-70 AB The linker for activation of T cells (LAT) is a critical adaptor molecule required for T cell antigen receptor (TCR)-mediated signaling and thymocyte development. Upon T cell activation, LAT becomes highly phosphorylated on tyrosine residues, and Grb2, Gads, and phospholipase C (PLC)-gamma 1 bind LAT via Src homology-2 domains. In LAT-deficient mutant Jurkat cells, TCR engagement fails to induce ERK activation, Ca2+ flux, and activation of AP-1 and NF-AT. We mapped the tyrosine residues in LAT responsible for interaction with these specific signaling molecules by expressing LAT mutants with tyrosine to phenylalanine mutations in LAT-deficient cells. Our results showed that three distal tyrosines, Tyr(171), Tyr(191), and Tyr(226), are responsible for Grb2-binding; Tyr171, and Tyr(191), but not Tyr(226) necessary for Gads binding. Mutation of Tyr(132) alone abolished PLC-gamma 1 binding. Mutation of all three distal tyrosines also abolished PLC-gamma 1 binding, suggesting there might be multiple binding sites for PLC-gamma 1. Mutation of Tyr(132) affected calcium flux and blocked Erk and NF-AT activation. Since Grb2 binding is not affected by this mutation, these results strongly suggest that PLC-gamma activation regulates Ras activation in these cells. Mutation of individual Grb2 binding sites had no functional effect, but mutation of two or three of these sites, in combination, also affected Erk. and NF-AT activation. C1 NCI, Div Basic Sci, Cellular & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Hosp Sick Children, Dept Med Biophys, Toronto, ON M5G 1X8, Canada. Hosp Sick Children, Arthur & Sonia Labatt Brain Tumour Res Ctr, Toronto, ON M5G 1X8, Canada. Duke Med Ctr, Dept Immunol, Durham, NC 27710 USA. RP Samelson, LE (reprint author), NCI, Div Basic Sci, Cellular & Mol Biol Lab, NIH, Bldg 37,Room 1E24,37 Convent Dr, Bethesda, MD 20892 USA. NR 30 TC 281 Z9 289 U1 0 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 28 PY 2000 VL 275 IS 30 BP 23355 EP 23361 DI 10.1074/jbc.M000404200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338KV UT WOS:000088419400104 PM 10811803 ER PT J AU Charles, V Mezey, E Reddy, PH Dehejia, A Young, TA Polymeropoulos, MH Brownstein, MJ Tagle, DA AF Charles, V Mezey, E Reddy, PH Dehejia, A Young, TA Polymeropoulos, MH Brownstein, MJ Tagle, DA TI Alpha-synuclein immunoreactivity of huntingtin polyglutamine aggregates in striatum and cortex of Huntington's disease patients and transgenic mouse models SO NEUROSCIENCE LETTERS LA English DT Article DE Huntington's disease; Parkinson's disease; immunohistochemistry; huntingtin; alpha-synuclein; transgenic mice; cerebral cortex; striatum ID SELECTIVE NEURONAL LOSS; LENGTH HD CDNA; PARKINSONS-DISEASE; MICE; GENE; LOCALIZATION; PLASTICITY; MUTATION; BRAIN AB Polyglutamine expansions in proteins are implicated in at least eight inherited neurodegenerative disorders, including Huntington's disease. These mutant proteins can form aggregates with in the nucleus and processes of neurons possibly due to misfolding of the proteins. Polyglutamine aggregates are ubiquitinated and sequester molecular chaperone proteins and proteasome components. To investigate other protein components of polyglutamine aggregates, cerebral cortex and striata from patients with Huntington's disease and full-length cDNA transgenic mouse models for this disease were examined immunohistochemically for alpha-synuclein reactivity. Our findings demonstrate that alpha-synuclein can be used as a marker for huntingtin polyglutamine aggregates in both human and mice. Moreover in the HD transgenic mice, the intensity of immunoreactivity increases with age. The significance of recruitment of alpha-synuclein into huntingtin aggregates and its translocation away from the synapses remains to be determined. We propose that aberrant interaction of mutant huntingtin with other proteins, including alpha-synuclein, may influence disease progression. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Natl Human Genome Res Inst, NIH, Bethesda, MD 20892 USA. NINDS, NIH, Bethesda, MD 20892 USA. NIMH, NIH, Bethesda, MD 20892 USA. RP Tagle, DA (reprint author), Bldg 49,Room 3A26,49 Convent Dr, Bethesda, MD 20892 USA. RI Brownstein, Michael/B-8609-2009 FU NIMH NIH HHS [MH/NS 31862] NR 17 TC 32 Z9 33 U1 0 U2 2 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 28 PY 2000 VL 289 IS 1 BP 29 EP 32 DI 10.1016/S0304-3940(00)01247-7 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 339EZ UT WOS:000088465800008 PM 10899401 ER PT J AU Hunyady, L Catt, KJ Clark, AJL Gaborik, Z AF Hunyady, L Catt, KJ Clark, AJL Gaborik, Z TI Mechanisms and functions of AT(1) angiotensin receptor internalization SO REGULATORY PEPTIDES LA English DT Review DE endocytosis; phosphorylation; dynamin; beta-arrestin; G protein-coupled receptor; Ca2+-mobilizing hormone ID PROTEIN-COUPLED RECEPTOR; MUSCARINIC ACETYLCHOLINE-RECEPTOR; AGONIST-INDUCED PHOSPHORYLATION; ADRENAL GLOMERULOSA CELLS; CLATHRIN-MEDIATED ENDOCYTOSIS; VASCULAR SMOOTH-MUSCLE; PLECKSTRIN HOMOLOGY DOMAIN; BETA-ADRENERGIC-RECEPTOR; II-BINDING PROTEIN; DYNAMIN-DEPENDENT ENDOCYTOSIS AB The type I (AT(1)) angiotensin receptor, which mediates the known physiological and pharmacological actions of angiotensin II, activates numerous intracellular signaling pathways and undergoes rapid internalization upon agonist binding. Morphological and biochemical studies have shown that agonist-induced endocytosis of the AT(1) receptor occurs via clathrin-coated pits, and is dependent on two regions in the cytoplasmic tail of the receptor. However, it is independent of G protein activation and signaling, and does not require the conserved NPXXY motif in the seventh transmembrane helix. The dependence of internalization of the AT(1) receptor on a cytoplasmic serine-threonine-rich region that is phosphorylated during agonist stimulation suggests that endocytosis is regulated by phosphorylation of the AT(1) receptor tail. beta-Arrestins have been implicated in the desensitization and endocytosis of several G protein-coupled receptors, but the exact nature of the adaptor protein required for association of the AT(1) receptor with clathrin-coated pits, and the role of dynamin in the internalization process, are still controversial. There is increasing evidence for a role of internalization in sustained signal generation from the AT(1) receptor. Several aspects of the mechanisms and specific function of AT(1) receptor internalization, including its precise mode and route of endocytosis, and the potential roles of cytoplasmic and nuclear receptors, remain to be elucidated. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Semmelweis Univ, Fac Med, Dept Physiol, H-1444 Budapest, Hungary. NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. St Bartholomews Hosp, Dept Endocrinol, London, England. St Bartholomews Hosp, Dept Chem Endocrinol, London, England. RP Hunyady, L (reprint author), Semmelweis Univ, Fac Med, Dept Physiol, POB 259, H-1444 Budapest, Hungary. EM hunyady@puskin.sote.hu NR 170 TC 68 Z9 72 U1 1 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-0115 J9 REGUL PEPTIDES JI Regul. Pept. PD JUL 28 PY 2000 VL 91 IS 1-3 BP 29 EP 44 DI 10.1016/S0167-0115(00)00137-3 PG 16 WC Endocrinology & Metabolism; Physiology SC Endocrinology & Metabolism; Physiology GA 354QW UT WOS:000089342000004 PM 10967200 ER PT J AU Ruda, MA Ling, QD Hohmann, AG Peng, YB Tachibana, T AF Ruda, MA Ling, QD Hohmann, AG Peng, YB Tachibana, T TI Altered nociceptive neuronal circuits after neonatal peripheral inflammation SO SCIENCE LA English DT Article ID SCIATIC-NERVE AFFERENTS; PAIN RESPONSE; DORSAL HORN; ADULT-RAT; SENSITIVITY; TERMINALS; PRETERM; SECTION AB Nociceptive neuronal circuits are formed during embryonic and postnatal times when painful stimuli are normally absent or Limited. Today, medical procedures for neonates with health risks can involve tissue injury and pain for which the Long-term effects are unknown. To investigate the impact of neonatal tissue injury and pain on development of nociceptive neuronal circuitry, we used an animal model of persistent hind paw peripheral inflammation, We found that, as adults, these animals exhibited spinal neuronal circuits with increased input and segmental changes in nociceptive primary afferent axons and altered responses to sensory stimulation. C1 Natl Inst Dent & Craniofacial Res, Cellular Neurosci Sect, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. RP Ruda, MA (reprint author), Natl Inst Dent & Craniofacial Res, Cellular Neurosci Sect, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. NR 16 TC 236 Z9 248 U1 3 U2 11 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 28 PY 2000 VL 289 IS 5479 BP 628 EP 630 DI 10.1126/science.289.5479.628 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 338XD UT WOS:000088447800056 PM 10915627 ER PT J AU Kaneko, S Hikida, T Watanabe, D Ichinose, H Nagatsu, T Kreitman, RJ Pastan, I Nakanishi, S AF Kaneko, S Hikida, T Watanabe, D Ichinose, H Nagatsu, T Kreitman, RJ Pastan, I Nakanishi, S TI Synaptic integration mediated by striatal cholinergic interneurons in basal ganglia function SO SCIENCE LA English DT Article ID DOPAMINE-RECEPTORS; STRIATOPALLIDAL NEURONS; GENE-EXPRESSION; MESSENGER-RNAS; MICE LACKING; RAT-BRAIN; D1; DISORDERS; LOCALIZATION; AMPHETAMINE AB The physiological role of striatal cholinergic interneurons was investigated with immunotoxin-mediated cell targeting (IMCT). Unilateral cholinergic cell ablation caused an acute abnormal turning behavior. These mice showed gradual recovery but displayed abnormal turning by both excess stimulation and inhibition of dopamine actions. In the acute phase, basal ganglia function was shifted to a hyperactive state by stimulation and suppression of striatonigral and striatopallidal neurons, respectively. D1 and D2 dopamine receptors were then down-regulated, relieving dopamine-predomimant synaptic perturbation but leaving a defect in controlling dopamine responses. The acetylcholine-dopamine interaction is concertedly and adaptively regulated for basal ganglia synaptic integration. C1 Kyoto Univ, Fac Med, Dept Biol Sci, Kyoto 6068501, Japan. Fujita Hlth Univ, Sch Med, Inst Comprehens Med Sci, Toyoake, Aichi 4701192, Japan. NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Kyoto Univ, Fac Med, Dept Biol Sci, Kyoto 6068501, Japan. EM snakanis@phy.med.kyoto-u.ac.jp RI Ichinose, Hiroshi/D-8986-2015 OI Ichinose, Hiroshi/0000-0001-6008-6860 NR 35 TC 122 Z9 126 U1 0 U2 3 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 EI 1095-9203 J9 SCIENCE JI Science PD JUL 28 PY 2000 VL 289 IS 5479 BP 633 EP 637 DI 10.1126/science.289.5479.633 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 338XD UT WOS:000088447800058 PM 10915629 ER PT J AU Howell, BW Herrick, TM Hildebrand, JD Zhang, YN Cooper, JA AF Howell, BW Herrick, TM Hildebrand, JD Zhang, YN Cooper, JA TI Dab1 tyrosine phosphorylation sites relay positional signals during mouse brain development SO CURRENT BIOLOGY LA English DT Article ID REELER-LIKE PHENOTYPE; SRC FAMILY KINASES; LAMINAR ORGANIZATION; NEURONAL MIGRATION; MUTANT MICE; PROTEIN; DISABLED-1; SCRAMBLER; RECEPTOR; ABL AB Background: The extracellular protein Rein controls neuronal migrations in parts of the cortex, hippocampus and cerebellum. In vivo, absence of Rein correlates with up-regulation of the docking protein Dab1 and decreased Dab1 tyrosine phosphorylation. Loss of the Rein receptor proteins, apolipoprotein receptor 2 and very low density lipoprotein receptor. results in a Rein-like phenotype accompanied by increased Dab1 protein expression, Complete loss of Dab1, however, recapitulates the Rein phenotype. Results: To determine whether Dab1 tyrosine phosphorylation affects Dab1 protein expression and positioning of embryonic neurons, we have identified Dab1 tyrosine phosphorylation sites. We then generated mice in which the Dab1 protein had all the potential tyrosine phosphorylation sites mutated. This mutant protein is not tyrosine phosphorylated during brain development and is not upregulated to the extent observed in the Rein or the apoER2 and VLDLR receptor mutants. Animals expressing the non-phosphorylated Dab1 protein have a phenotype similar to the dab1-null mutant. Conclusions: Dab1 is downregulated by the Rein signal in neurons in the absence of tyrosine phosphorylation. Dab1 tyrosine phosphorylation sites and not downregulation of Dab1 protein are required for Rein signaling. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA. NINDS, Neurogenet Branch, NIH, Bethesda, MD 20892 USA. RP Howell, BW (reprint author), Fred Hutchinson Canc Res Ctr, 1100 Fairview Ave N, Seattle, WA 98109 USA. OI Howell, Brian/0000-0002-0204-0773 FU NCI NIH HHS [5P30-CA15704-26, R37-CA41072] NR 53 TC 197 Z9 207 U1 1 U2 3 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 27 PY 2000 VL 10 IS 15 BP 877 EP 885 DI 10.1016/S0960-9822(00)00608-4 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 348HK UT WOS:000088979300015 PM 10959835 ER PT J AU Paull, TT Rogakou, EP Yamazaki, V Kirchgessner, CU Gellert, M Bonner, WM AF Paull, TT Rogakou, EP Yamazaki, V Kirchgessner, CU Gellert, M Bonner, WM TI A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage SO CURRENT BIOLOGY LA English DT Article ID DOUBLE-STRAND BREAKS; DEPENDENT PROTEIN-KINASE; ATAXIA-TELANGIECTASIA GENE; HUMAN TUMOR-CELLS; SACCHAROMYCES-CEREVISIAE; PHOSPHATIDYLINOSITOL 3-KINASE; IONIZING-RADIATION; CELLULAR DNA; CHO CELLS; RECOMBINATION AB Background: The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. Recently it has been reported that a member of the histone H2A family, H2AX, becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites. Results: In this work, we examine the role of H2AX phosphorylation in focus formation by several repair-related complexes, and investigate what factors may be involved in initiating this response. Using two different methods to create DNA double-strand breaks in human cells, we found that the repair factors Rad50 and Rad51 each colocalized with phosphorylated H2AX (gamma-H2AX) foci after DNA damage. The product of the tumor suppressor gene BRCA1 also colocalized with gamma-H2AX and was recruited to these sites before Rad50 or Rad51. Exposure of cells to the fungal inhibitor wortmannin eliminated focus formation by all repair factors examined, suggesting a role for the phosphoinositide (PI)-3 family of protein kinases in mediating this response. Wortmannin treatment was effective only when it was added early enough to prevent gamma-H2AX formation, indicating that gamma-H2AX is necessary for the recruitment of other factors to the sites of DNA damage. DNA repair-deficient cells exhibit a substantially reduced ability to increase the phosphorylation of H2AX in response to ionizing radiation, consistent with a role for gamma-H2AX in DNA repair. Conclusions: The pattern of gamma-H2AX fool that is established within a few minutes of DNA damage accounts for the patterns of Rad50, Rad51, and Brca1 foci seen much later during recovery from damage. The evidence presented strongly supports a role for the gamma-H2AX and the PI-3 protein kinase family in focus formation at sites of double-strand breaks and suggests the possibility of a change in chromatin structure accompanying double-strand break repair. (C) 2000 Elsevier Science Ltd, All rights reserved. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Stanford Univ, Dept Radiat Oncol, Sch Med, Mayer Canc Res Lab, Stanford, CA 94305 USA. RP Gellert, M (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RI qiao, zhixin/I-3408-2012 NR 62 TC 1295 Z9 1346 U1 8 U2 64 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 27 PY 2000 VL 10 IS 15 BP 886 EP 895 DI 10.1016/S0960-9822(00)00610-2 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 348HK UT WOS:000088979300016 PM 10959836 ER PT J AU Shevach, EM AF Shevach, EM TI Suppressor T cells: Rebirth, function and homeostasis SO CURRENT BIOLOGY LA English DT Article AB A minor subpopulation of CD4(+) T cells that co express the cell-surface molecule CD25 has emerged as the major population of T cells responsible for downregulation of the immune response and prevention of autoimmunity. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Shevach, EM (reprint author), NIAID, Immunol Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 12 TC 27 Z9 30 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 27 PY 2000 VL 10 IS 15 BP R572 EP R575 DI 10.1016/S0960-9822(00)00617-5 PG 4 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 348HK UT WOS:000088979300014 PM 10959833 ER PT J AU Arris, CE Boyle, FT Calvert, AH Curtin, NJ Endicott, JA Garman, EF Gibson, AE Golding, BT Grant, S Griffin, RJ Jewsbury, P Johnson, LN Lawrie, AM Newell, DR Noble, MEM Sausville, EA Schultz, R Yu, W AF Arris, CE Boyle, FT Calvert, AH Curtin, NJ Endicott, JA Garman, EF Gibson, AE Golding, BT Grant, S Griffin, RJ Jewsbury, P Johnson, LN Lawrie, AM Newell, DR Noble, MEM Sausville, EA Schultz, R Yu, W TI Identification of novel purine and pyrimidine cyclin-dependent kinase inhibitors with distinct molecular interactions and tumor cell growth inhibition profiles SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID 2,6,9-TRISUBSTITUTED PURINES; CRYSTAL-STRUCTURE; CARCINOMA-CELLS; CDK INHIBITORS; PROTEIN MODELS; FLAVOPIRIDOL; POTENT; APOPTOSIS; CANCER; ANALOGS AB Substituted guanines and pyrimidines were tested as inhibitors of cyclin B1/CDK1 and cyclin A3/CDK2 and soaked into crystals of monomeric CDK2. O-6-Cyclohexylmethylguanine (NU2058) was a competitive inhibitor of CDK1 and CDK2 with respect to ATP (Ki values: CDK1, 5 +/-: 1 mu M; CDK2, 12 +/- 3 mu M) and formed a triplet of hydrogen bonds (i.e., NH-9 to Glu 81, N-3 to Leu 83, and 2-NH2 to Leu 83). The triplet of hydrogen bonding and CDK inhibition was reproduced by 2,6-diamino-4-cyclohexylmethyloxy-5-nitrosopyrimidine (NU6027, K-i values: CDK1, 2.5 +/- 0.4 mu M; CDK2, 1.3 +/- 0.2 mu M). Against human tumor cells, NU2058 and NU6027 were growth inhibitory in vitro (mean GI(50) values of 13 +/- 7 mu M and 10 +/- 6 mu M, respectively), with a pattern of sensitivity distinct from flavopiridol and olomoucine. These CDK inhibition and chemosensitivity data indicate that the distinct mode of binding of NU2058 and NU6027 has direct consequences for enzyme and cell growth inhibition. C1 Univ Newcastle Upon Tyne, Sch Med, Canc Res Unit, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. Univ Newcastle Upon Tyne, Sch Med, Dept Chem, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. AstraZeneca, Alderley Pk, Cheshire, England. Univ Oxford, Mol Biophys Lab, Oxford OX1 3QU, England. Univ Oxford, Dept Biochem, Oxford OX1 3QU, England. NCI, Dev Therapeut Program, Bethesda, MD 20892 USA. RP Newell, DR (reprint author), Univ Newcastle Upon Tyne, Sch Med, Canc Res Unit, Framlington Pl, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. OI Noble, Martin/0000-0002-3595-9807 NR 46 TC 176 Z9 182 U1 2 U2 21 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 27 PY 2000 VL 43 IS 15 BP 2797 EP 2804 DI 10.1021/jm990628o PG 8 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 339DL UT WOS:000088462300005 PM 10956187 ER PT J AU Rick, SW AF Rick, SW TI Free energy, entropy and heat capacity of the hydrophobic interaction as a function of pressure SO JOURNAL OF PHYSICAL CHEMISTRY B LA English DT Article ID MOLECULAR-DYNAMICS SIMULATIONS; CHARGE FORCE-FIELDS; LIQUID WATER; MONTE-CARLO; TEMPERATURE-DEPENDENCE; SOLVENT POLARIZABILITY; COMPUTER-SIMULATIONS; POTENTIAL FUNCTIONS; AQUEOUS-SOLUTION; RIBONUCLEASE-A AB Molecular dynamics simulations of a methane pair in water are used to calculate the thermodynamic properties of the hydrophobic interaction as a function of pressure. Pressure is found to decrease the tendency to form methane aggregates. The entropic contribution to the free energy, which at atmospheric pressure greatly stabilizes aggregation, is highly pressure dependent. As the pressure increases, the entropic stabilization steadily decreases until, at 7 kbar, the entropy of the contact pair is equal to the entropy of the solvent separated pair. The heat capacity change between the contact and solvent separated Fair is shown to be large and positive at 1 atm, as is characteristic of hydrophobic processes. At higher pressures, the heat capacity change is zero, indicating that two of the significant properties of the hydrophobic effect, the large entropy decrease and heat capacity increase are lost at high pressures. The free energy, volume and entropy changes are consistent with the corresponding changes for the pressure denaturation of proteins. C1 NCI, Adv Biomed Comp Ctr, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD USA. RP Univ New Orleans, Dept Chem, New Orleans, LA 70148 USA. NR 57 TC 54 Z9 54 U1 0 U2 6 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 1520-6106 J9 J PHYS CHEM B JI J. Phys. Chem. B PD JUL 27 PY 2000 VL 104 IS 29 BP 6884 EP 6888 DI 10.1021/jp000841s PG 5 WC Chemistry, Physical SC Chemistry GA 339QY UT WOS:000088490500027 ER PT J AU Schneider, TD AF Schneider, TD TI The bottle - Force the weird stuff down SO NATURE LA English DT Article C1 NIH, Frederick, MD USA. RP Schneider, TD (reprint author), NIH, Frederick, MD USA. NR 0 TC 0 Z9 0 U1 1 U2 5 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 27 PY 2000 VL 406 IS 6794 BP 351 EP 351 DI 10.1038/35019183 PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 337WC UT WOS:000088383800022 ER PT J AU McKay, R AF McKay, R TI Stem cells - hype and hope SO NATURE LA English DT Article ID TRANSPLANTATION; MOUSE; DERIVATION; PRECURSORS C1 NINDS, NIH, LMB, Bethesda, MD 20892 USA. RP McKay, R (reprint author), NINDS, NIH, LMB, Bethesda, MD 20892 USA. NR 25 TC 167 Z9 177 U1 0 U2 15 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 27 PY 2000 VL 406 IS 6794 BP 361 EP 364 DI 10.1038/35019186 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 337WC UT WOS:000088383800029 PM 10935622 ER PT J AU Lieu, T Ray, GT Black, S Shinefield, H Butler, J Miller, M AF Lieu, T Ray, GT Black, S Shinefield, H Butler, J Miller, M TI Cost-effectiveness of pneumococcal vaccine - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter C1 Harvard Pilgrim Hlth Care, Boston, MA USA. Kaiser Permanente, Oakland, CA USA. Ctr Dis Control & Prevent, Atlanta, GA USA. NIH, Bethesda, MD 20892 USA. RP Lieu, T (reprint author), Harvard Pilgrim Hlth Care, Boston, MA USA. NR 2 TC 5 Z9 6 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 26 PY 2000 VL 284 IS 4 BP 440 EP 441 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 335QB UT WOS:000088254200023 PM 10904501 ER PT J AU Perry, HM Davis, BR Price, TR Applegate, WB Fields, WS Guralnik, JM Kuller, L Pressel, S Stamler, J Probstfield, JL AF Perry, HM Davis, BR Price, TR Applegate, WB Fields, WS Guralnik, JM Kuller, L Pressel, S Stamler, J Probstfield, JL CA Systolic Hypertension Elderly Prog TI Effect of treating isolated systolic hypertension on the risk of developing various types and subtypes of stroke - The Systolic Hypertension in the Elderly Program (SHEP) SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HEMORRHAGE AB Context The Systolic Hypertension in the Elderly Program (SHEP) demonstrated that treating isolated systolic hypertension in older patients decreased incidence of total stroke, but whether all types of stroke were reduced was not evaluated. Objective To investigate antihypertensive drug treatment effects on incidence of stroke by type and subtype, timing of strokes, case-fatality rates, stroke residual effects, and relationship of attained systolic blood pressure to stroke incidence. Design The SHEP study, a randomized, double-blind, placebo-controlled trial began March 1, 1985, and had an average follow-up of 4.5 years. Setting and Participants A total of 4736 men and women aged 60 years or older with isolated systolic hypertension at 16 clinical centers in the United States. Interventions Patients were randomly assigned to receive treatment with 12.5 mg/d of chlorthalidone (step 1); either 25 mg/d of atenolol or 0.05 mg/d of reserpine (step 2) could be added (n = 2365); or placebo (n = 2371). Main Outcome Measures Occurrence, type and subtype, and timing of first strokes and stroke fatalities; and change in stroke incidence for participants (whether in active treatment or placebo groups) reaching study-specific systolic blood pressure goal (decrease of at least 20 mm Hg from baseline to below 160 mm Hg) compared with participants not reaching goal. Results A total of 85 and 132 participants in the active treatment and placebo groups, respectively, had ischemic strokes (adjusted relative risk [RR], 0.63; 95% confidence interval [CI], 0.48-0.82); 9 and 19 had hemorrhagic strokes (adjusted RR, 0.46; 95% CI, 0.21-1.02); and 9 and 8 had strokes of unknown type (adjusted RR, 1.05; 95% CI, 0.40-2.73), respectively. Four subtypes of ischemic stroke were observed in active treatment and placebo group participants, respectively, as follows: for lacunar, n=23 and n=43 (adjusted RR, 0.53; 95% CI, 0.32-0.88); for embolic, n=9 and n=16 (adjusted RR, 0.56; 95% CI, 0.25-1.27); for atherosclerotic, n=13 and n=13 (adjusted RR, 0.99; 95% CI, 0.46-2.15); and for unknown subtype, n=40 and n=60 (adjusted RR, 0.64; 95% CI, 0.43-0.96). Treatment effect was observed within 1 year for hemorrhagic strokes but was not seen until the second year for ischemic strokes. Stroke incidence significantly decreased in participants attaining study-specific systolic blood pressure goals. Conclusions In this study, antihypertensive drug treatment reduced the incidence of both hemorrhagic and ischemic (including lacunar) strokes. Reduction in stroke incidence occurred when specific systolic blood pressure goals were attained. C1 Washington Univ, Dept Med, St Louis, MO 63110 USA. Univ Texas, Hlth Sci Ctr, Sch Publ Hlth, Houston, TX USA. Univ Maryland, Sch Med, Dept Neurol, Baltimore, MD 21201 USA. Wake Forest Univ, Dept Med, Winston Salem, NC 27109 USA. NIA, Bethesda, MD 20892 USA. Univ Pittsburgh, Dept Epidemiol, Pittsburgh, PA 15261 USA. Northwestern Univ, Sch Med, Dept Prevent Med, Chicago, IL USA. Univ Washington, Dept Med, Seattle, WA USA. RP Perry, HM (reprint author), Washington Univ, Dept Med, Box 8048,660 S Euclid Ave, St Louis, MO 63110 USA. NR 21 TC 161 Z9 168 U1 0 U2 10 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 26 PY 2000 VL 284 IS 4 BP 465 EP 471 DI 10.1001/jama.284.4.465 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 335QB UT WOS:000088254200033 PM 10904510 ER PT J AU Topol, IA Nemukhin, AV Chao, M Iyer, LK Tawa, GJ Burt, SK AF Topol, IA Nemukhin, AV Chao, M Iyer, LK Tawa, GJ Burt, SK TI Quantum chemical studies of reactions of the cyclic disulfides with the zinc finger domains in the HIV-1 nucleocapsid protein (NCp7) SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID DENSITY-FUNCTIONAL THEORY; BOUNDARY-ELEMENT METHOD; SOLVATION FREE-ENERGY; MOLECULAR ELECTROSTATICS; DIELECTRIC MODEL; CONTINUUM MODEL; EXACT EXCHANGE; AB-INITIO; INHIBITORS; SOLVENT AB By using quantum chemistry methods, including ab initio Hartree-Fock (HF), as well as the Density Functional Theory approach employing B3LYP approximation, the reaction profiles of three cyclic disulfide species with model zinc finger domains in the HIV-1 nucleocapsid protein (NCp7) have been analyzed. It is shown that the disulfide molecules can act as efficient agents destroying the tetrahedral coordination sphere of the zinc finger domains. The consequence of the reaction is a break of one of the Zn-S bonds and removal of the corresponding molecular thiolate group from the domain by forming a new S-S bond between sulfur atoms from the withdrawing fragment and of the electrophilic agent. As a result of this process the zinc-containing site transforms from the initial tetrahedral geometry to a planar geometry configuration. This transformation further facilitates destruction of the metal binding site. The calculations explicitly show correlations between redox potentials of the electrophilic agents, their reactivity, and barrier heights on the reaction pathway. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. Moscow MV Lomonosov State Univ, Dept Chem, Moscow 119899, Russia. RP Topol, IA (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Adv Biomed Comp Ctr, POB B, Frederick, MD 21702 USA. RI Nemukhin, Alexander/P-9662-2015 NR 48 TC 11 Z9 11 U1 0 U2 4 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 26 PY 2000 VL 122 IS 29 BP 7087 EP 7094 DI 10.1021/ja000585x PG 8 WC Chemistry, Multidisciplinary SC Chemistry GA 340FP UT WOS:000088522700032 ER PT J AU Lenfant, C AF Lenfant, C TI Training the next generation of biomedical researchers - Challenges and opportunities SO CIRCULATION LA English DT Editorial Material DE editorials; research training; mentorship; diversity; clinical research 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 11 Z9 11 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 25 PY 2000 VL 102 IS 4 BP 368 EP 370 PG 3 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 337RA UT WOS:000088374400009 PM 10908205 ER PT J AU Nelson, SA Santora, KE LaRochelle, WJ AF Nelson, SA Santora, KE LaRochelle, WJ TI Isolation and characterization of a novel PDGF-induced human gene SO GENE LA English DT Article DE NOP5/NOP58; PDGF; ribosome biogenesis ID PRE-RIBOSOMAL-RNA; YEAST; EXPRESSION; CLEAVAGE; CLONING; SITE AB Using a differential display RT-PCR strategy to identify novel growth-factor-induced transcripts, we cloned and characterized the human homolog of yeast NOP5/NOP58, whose gene product has been implicated in the execution of early pre-rRNA processing steps. Human NOP5 cDNA was isolated from an M426 fibroblast cDNA library. Determination of the cDNA nucleotide sequence revealed an open reading frame of 1587 nucleotides encoding a predicted gene product of 529 amino acids and mass of 59 554 Da. The yeast and human NOP5 gene products were found to share 63% homology and 46% identity. NOP5 mRNA was induced within 2 h of platelet-derived growth factor (PDGF) treatment of human M426 fibroblasts. Pretreatment with cycloheximide enhanced, while actinomycin blocked induction of the NOP5 transcript. In vitro translational analysis of the cDNA revealed a 60 kDa species? consistent with the predicted molecular weight of the gene product. Ubiquitous, but differential NOP5 mRNA expression was revealed after Northern blot analysis of total RNA from several human tissues. Moreover, NOP5 mRNA expression was also demonstrated in cell lines of fibroblast, epithelial, and myeloid origin. A highly charged carboxy terminal domain and consensus phosphorylation sites were identified. The presence of potential regulatory elements, together with growth factor induction and widespread expression is consistent with the hypothesis that the NOP5 gene product may play a role in fundamental cellular growth processes. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Cellular & Mol Biol Lab, Bethesda, MD 20892 USA. RP LaRochelle, WJ (reprint author), NCI, Cellular & Mol Biol Lab, Bldg 37,Room 1E24, Bethesda, MD 20892 USA. NR 24 TC 3 Z9 4 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD JUL 25 PY 2000 VL 253 IS 1 BP 87 EP 93 DI 10.1016/S0378-1119(00)00232-8 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 344EE UT WOS:000088744000009 PM 10925205 ER PT J AU Haidan, A Bornstein, SR Liu, Z Walsh, LP Stocco, DM Ehrhart-Bornstein, M AF Haidan, A Bornstein, SR Liu, Z Walsh, LP Stocco, DM Ehrhart-Bornstein, M TI Expression of adrenocortical steroidogenic acute regulatory (StAR) protein is influenced by chromaffin cells SO MOLECULAR AND CELLULAR ENDOCRINOLOGY LA English DT Article DE steroidogenic acute regulatory protein; peripheral benzodiazepine receptor; adrenocortical cells; chromaffin cells; steroidogenesis, (bovine adrenal) ID MESSENGER-RIBONUCLEIC-ACID; ATRIAL-NATRIURETIC-PEPTIDE; ADRENAL-CORTEX CELLS; SIDE-CHAIN CLEAVAGE; IN-VIVO; FASCICULATA-RETICULARIS; BENZODIAZEPINE RECEPTOR; LUTEINIZING-HORMONE; LEYDIG-CELLS; CHOLESTEROL AB Cell-cell interactions are crucial role for the proper functioning of endocrine glands. We recently demonstrated that interactions of chromaffin and cortical cells are important for adrenocortical steroidogenesis. However, the molecular mechanisms have not been elucidated and it is unclear if this involves acute and/or chronic processes. By Northern analysis and the quantitative technique of TaqMan PCR we investigated whether chromaffin cells influence the regulation of StAR and the peripheral benzodiazepine receptor (PBR), both required for the rate-limiting step, the delivery of cholesterol to the inner mitochondrial membrane. StAR mRNA levels in bovine adrenocortical cells were increased by incubation with chromaffin cell-conditioned medium (CCM). Short-term treatment for 4 h resulted in a greater stimulation (229 +/- 29% of basal, mean +/- SEM) than did longer incubation times of 8 h and 5 days (159 +/- 13 and 177 +/- 24%). Neither short nor a long-term treatment affected PER expression. Consistently, the major secretion of chromaffin cells, epinephrine dose-dependently stimulated StAR expression with no effect on PER mRNA. In conclusion, adrenomedullary secretory products are not necessary for the maintenance of PER expression but facilitate steroid biosynthesis by increasing StAR mRNA expression and therefore can account for an ACTH-independent regulation of the rate-limiting step in steroidogenesis. (C) 2000 Published by Elsevier Science Ireland Ltd. All rights reserved. C1 NICHD, NIH, Bethesda, MD 20892 USA. Univ Leipzig, Dept Internal Med 3, A-04103 Leipzig, Austria. NIMH, NIH, Bethesda, MD 20892 USA. Texas Tech Univ, Hlth Sci Ctr, Dept Cell Biol & Biochem, Lubbock, TX 79430 USA. RP Bornstein, SR (reprint author), NICHD, NIH, Room 10N262,Bldg 10,10 Ctr Dr, Bethesda, MD 20892 USA. FU NICHD NIH HHS [HD17481] NR 48 TC 18 Z9 18 U1 0 U2 1 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 25 PY 2000 VL 165 IS 1-2 BP 25 EP 32 DI 10.1016/S0303-7207(00)00267-7 PG 8 WC Cell Biology; Endocrinology & Metabolism SC Cell Biology; Endocrinology & Metabolism GA 346AT UT WOS:000088848500004 PM 10940480 ER PT J AU Song, S Adachi, K Katsuyama, M Sorimachi, K Oka, T AF Song, S Adachi, K Katsuyama, M Sorimachi, K Oka, T TI Isolation and characterization of the 5 '-upstream and untranslated regions of the mouse type II iodothyronine deiodinase gene SO MOLECULAR AND CELLULAR ENDOCRINOLOGY LA English DT Article DE genomic organization; open reading frame; promoter activity; transcription start site; type II iodothyronine deiodinase ID THYROID-HORMONE ACTION; MESSENGER-RIBONUCLEIC-ACID; 5'-DEIODINASE GENE; CLONING; EXPRESSION; CDNA; CELLS; SELENOPROTEIN; TRANSLATION; DEFICIENCY AB The type II iodothyronine deiodinase (D2) catalyzes the 5'-deiodinationc of thyroxine to yield the biologically active form, 3,3',5,-tri-iodothyronine, and is a member of the selenoproteins. We isolated a 17.5 kb mouse genomic clone containing the entire coding and 5'-untranslated regions of the D2 gene (mdio2). We also isolated the entire 5'-UTR of the mouse D2 cDNA, which was 753 bp in length and contained five ATG codons. An additional 258 bp ORF from the fourth ATG codon was found in the same reading frame as the coding region reported previously, and this additional ORF contained a TGA codon, which could encode selenocysteine. The proximal promoter of mdio2 contained a TATA box and several potential transcription factor-binding sequences, including CRE, C/EBP and GATA binding sites. The 1.3 kb 5'-upstream region exhibited a promoter activity by reporter assay using Mm5MT and JAR cells, which have a D2 transcript, but not HepG2 cells that have no detectable level of D2 transcript. Published by Elsevier Science Ireland Ltd. C1 NIDDK, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. RP Oka, T (reprint author), NIDDK, Lab Genet & Physiol, NIH, Room 106,Bldg 8,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Sorimachi, Kenji/0000-0002-5259-9671 NR 39 TC 22 Z9 23 U1 0 U2 0 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 25 PY 2000 VL 165 IS 1-2 BP 189 EP 198 DI 10.1016/S0303-7207(00)00246-X PG 10 WC Cell Biology; Endocrinology & Metabolism SC Cell Biology; Endocrinology & Metabolism GA 346AT UT WOS:000088848500021 PM 10940497 ER PT J AU Van Houten, B Cheng, S Chen, YM AF Van Houten, B Cheng, S Chen, YM TI Measuring gene-specific nucleotide excision repair in human cells using quantitative amplification of long targets from nanogram quantities of DNA SO MUTATION RESEARCH-DNA REPAIR LA English DT Article ID COMPLEMENTATION GROUP-C; HAMSTER OVARY CELLS; INTERSTRAND CROSS-LINKS; CHAIN-REACTION ASSAY; DIHYDROFOLATE-REDUCTASE; SEQUENCE SPECIFICITY; PREFERENTIAL REPAIR; PCR AMPLIFICATION; PYRIMIDINE DIMERS; HUMAN FIBROBLASTS AB We have been developing a rapid and convenient assay for the measurement of DNA damage and repair in specific genes using quantitative polymerase chain reaction (QPCR) methodology. Since the sensitivity of this assay is limited to the size of the DNA amplification fragment, conditions have been found for the quantitative generation of PCR fragments from human genomic DNA in the range of 6-24 kb in length. These fragments include: (1) a 16.2 kb product from the mitochondrial genome; (2) 6.2, 10.4 kb, and 15.4 kb products from the hprt gene, and (3) 13.5, 17.7, 24.2 kb products from the human beta-globin gene cluster. Exposure of SV40 transformed human fibroblasts to increasing fluences of ultraviolet light (UV) resulted in the linear production of photoproducts with 10 J/m(2) of UVC producing 0.085 and 0.079 lesions/kb in the hprt gene and the beta-globin gene cluster, respectively. Kinetic analysis of repair following 10 J/m(2) of WC exposure indicated that the time necessary for the removal of 50% of the photoproducts, in the hprt gene and beta-globin gene cluster was 7.8 and 24.2 h, respectively. Studies using lymphoblastoid cell lines show very little repair in XPA cells in both the hprt gene and beta-globin locus. Preferential repair in the hprt gene was detected in XPC cells. Cisplatin lesions were also detected using this method and showed slower rates of repair than UV-induced photoproducts. These data indicate that the use of long targets in the gene-specific QPCR assay allows the measurement of biologically relevant lesion frequencies in 5-30 ng of genomic DNA. This assay will be useful for the measurement of human exposure to genotoxic agents and the determination of human repair capacity. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77550 USA. Roche Mol Syst, Alameda, CA USA. RP Van Houten, B (reprint author), NIEHS, Mol Genet Lab, DERT, 111 TW Alexander Dr,Box 12233, Res Triangle Pk, NC 27709 USA. FU NIEHS NIH HHS [1P30ES06676-01A1, 1RO1ES07038-02, 1RO1ES07218-01] NR 37 TC 35 Z9 35 U1 1 U2 5 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0921-8777 J9 MUTAT RES-DNA REPAIR JI Mutat. Res.-DNA Repair PD JUL 25 PY 2000 VL 460 IS 2 BP 81 EP 94 DI 10.1016/S0921-8777(00)00018-5 PG 14 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 334LJ UT WOS:000088187700001 PM 10882849 ER PT J AU Peng, A Koffman, BM Malley, JD Dalakas, MC AF Peng, A Koffman, BM Malley, JD Dalakas, MC TI Disease progression in sporadic inclusion body myositis: Observations in 78 patients SO NEUROLOGY LA English DT Article AB Functional decline for each decade at symptom onset and need for cane, walker, or wheelchair were assessed in 78 biopsy-proved patients with sporadic inclusion body myositis. Patients with disease onset between 40 and 59 years used a walker after 10.2 +/- 5.8 years, whereas those with disease onset between 60 and 79 years used a walker after 5.7 +/- 5.0 years (p = 0.05). Because patients progress faster to disability when symptoms begin after the age of 60, age at disease onset may define patient subsets for stratification in clinical trials. C1 NINDS, Neuromuscular Dis Sect, NIH, Bethesda, MD 20892 USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. RP Dalakas, MC (reprint author), NINDS, Neuromuscular Dis Sect, NIH, 10 Ctr Dr,MSC 1382, Bethesda, MD 20892 USA. NR 10 TC 50 Z9 51 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 25 PY 2000 VL 55 IS 2 BP 296 EP 298 PG 3 WC Clinical Neurology SC Neurosciences & Neurology GA 337FA UT WOS:000088345700026 PM 10908910 ER PT J AU Moffat, SD Zonderman, AB Harman, SM Blackman, MR Kawas, C Resnick, SM AF Moffat, SD Zonderman, AB Harman, SM Blackman, MR Kawas, C Resnick, SM TI The relationship between longitudinal declines in dehydroepiandrosterone sulfate concentrations and cognitive performance in older men SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID ALZHEIMERS-DISEASE; ELDERLY WOMEN; DHEA; MEMORY; BRAIN; AGE; CORTISOL; YOUTH AB Background: The observation that dehydroepiandrosterone (DHEA) concentrations decrease markedly with age has led to the hypothesis that declining DHEA concentrations may contribute to age-related changes in cognition. In the United States, DHEA is widely available as an over-the-counter supplement that individuals are using in an effort to ameliorate age-related cognitive and physical changes. Objective: To investigate the relationship between age-associated decreases in endogenous DHEA sulfate (DHEA-S) concentrations and declines in neuropsychological performance in a prospective, longitudinal study. Methods: The subjects were 883 men from a community-dwelling volunteer sample in the Baltimore Longitudinal Study of Aging. The men were aged 22 to 91 years at the initial visit, and they were followed up for as long as 31 years (mean, 11.55 years), with biennial reassessments of multiple cognitive domains and contemporaneous measurement of serum DHEA-S concentrations. Outcome measures were the results of cognitive tests of verbal and visual memory, 2 tests of mental status, phonemic and semantic word fluency tests, and measures of visuomotor scanning and attention. Serum DHEA-S concentrations were determined by standard radioimmunoassay. Results: Neither the rates of decline in mean DHEA-S concentrations nor the mean DHEA-S concentrations within individuals were related to cognitive status or cognitive decline. A comparison between the highest and lowest DHEA-S quartiles revealed no cognitive differences, despite the fact that these groups differed in endogenous DHEA-S concentration by more than a factor of 4 for a mean duration of 12 years. Conclusion: Our longitudinal results augment those of previous prospective studies by suggesting that the decline in endogenous DHEA-S concentration is independent of cognitive status and cognitive decline in healthy aging men. C1 NIA, Lab Personal & Cognit, Baltimore, MD 21224 USA. NIA, Clin Invest Lab, Intramural Res Program, Baltimore, MD 21224 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Neurol, Baltimore, MD 21205 USA. RP Moffat, SD (reprint author), NIA, Lab Personal & Cognit, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Zonderman, Alan B/0000-0002-6523-4778 FU NIA NIH HHS [AG05146, AG08325] NR 31 TC 44 Z9 50 U1 1 U2 2 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 24 PY 2000 VL 160 IS 14 BP 2193 EP 2198 DI 10.1001/archinte.160.14.2193 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 335FM UT WOS:000088233300016 PM 10904463 ER PT J AU Ghatan, S Larner, S Kinoshita, Y Hetman, M Patel, L Xia, ZG Youle, RJ Morrison, RS AF Ghatan, S Larner, S Kinoshita, Y Hetman, M Patel, L Xia, ZG Youle, RJ Morrison, RS TI p38 MAP kinase mediates Bax translocation in nitric oxide-induced apoptosis in neurons SO JOURNAL OF CELL BIOLOGY LA English DT Review DE caspase; excitotoxicity; neuronal cell death; p53; mitochondria ID ACTIVATED PROTEIN-KINASE; N-TERMINAL KINASE; CYTOCHROME-C RELEASE; TUMOR-NECROSIS-FACTOR; PERMEABILITY TRANSITION PORE; CEREBELLAR GRANULE CELLS; NMDA RECEPTOR ACTIVATION; DEFICIENT MICE; SIGNAL-TRANSDUCTION; MAMMALIAN-CELLS AB Nitric oxide is a chemical messenger implicated in neuronal damage associated with ischemia, neurodegenerative disease, and excitotoxicity. Excitotoxic injury leads to increased NO formation, as well as stimulation of the p38 mitogen-activated protein (MAP) kinase in neurons. In the present study, we determined if NO-induced cell death in neurons was dependent on p38 MAP kinase activity. Sodium nitroprusside (SNP), an NO donor, elevated caspase activity and induced death in human SH-SY5Y neuroblastoma cells and primary cultures of cortical neurons. Concomitant treatment with SB203580, a p38 MAP kinase inhibitor, diminished caspase induction and protected SH-SY5Y cells and primary cultures of cortical neurons from NO-induced cell death, whereas the caspase inhibitor zVAD-fmk did not provide significant protection. A role for p38 MAP kinase was further substantiated by the observation that SB203580 blocked translocation of the cell death activator, Bax, from the cytosol to the mitochondria after treatment with SNP. Moreover, expressing a constitutively active form of MKK3, a direct activator of p38 MAP kinase promoted Bax translocation and cell death in the absence of SNP. Bax-deficient cortical neurons were resistant to SNP, further demonstrating the necessity of Bax in this mode of cell death. These results demonstrate that p38 MAP kinase activity plays a critical role in NO-mediated cell death in neurons by stimulating Bax translocation to the mitochondria, thereby activating the cell death pathway. C1 Univ Washington, Sch Med, Dept Neurol Surg, Seattle, WA 98195 USA. Univ Washington, Sch Publ Hlth, Dept Environm Hlth, Seattle, WA 98195 USA. NINDS, Biochem Sect, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Morrison, RS (reprint author), Univ Washington, Sch Med, Dept Neurol Surg, Box 356470, Seattle, WA 98195 USA. FU NIA NIH HHS [AG10917]; NINDS NIH HHS [R01 NS035533, NS35533, NS37359] NR 100 TC 316 Z9 340 U1 0 U2 10 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP 335 EP 347 DI 10.1083/jcb.150.2.335 PG 13 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400017 PM 10908576 ER PT J AU Watts, NR Sackett, DL Ward, RD Miller, MW Wingfield, PT Stahl, SS Steven, AC AF Watts, NR Sackett, DL Ward, RD Miller, MW Wingfield, PT Stahl, SS Steven, AC TI HIV-1 Rev depolymerizes microtubules to form stable bilayered rings SO JOURNAL OF CELL BIOLOGY LA English DT Article DE acquired immunodeficiency syndrome; HIV-1 Rev; kinesin; microtubules; tubulin ID VIRUS TYPE-1 REV; TUBULIN DIMER; SUBCELLULAR-LOCALIZATION; NUCLEAR EXPORT; MESSENGER-RNAS; NEF PROTEIN; IN-VITRO; BINDING; ASSOCIATION; PHOSPHORYLATION AB We describe a novel interaction between HIV-1 Rev and microtubules (MTs) that results in the formation of bilayered rings that are 44-49 nm in external diameter, 3.4-4.2 MD (megadaltons) in mass, and have 28-, 30-, or 32-fold symmetry. Ring formation is not sensitive to taxol, colchicine, or microtubule-associated proteins, but requires Mg2+ and is inhibited by maytansine. The interaction involves the NH2-terminal domain of Rev and the face of tubulin exposed on the exterior of the MTs. The NH2-terminal half of Rev has unexpected sequence similarity to the tubulin-binding portion of the catalytic/motor domains of the microtubule-destabilizing Kin I kinesins. We propose a model wherein binding of Rev dimers to MTs at their ends causes segments of two neighboring protofilaments to peel off and close into rings, circumferentially containing 14, 15, or 16 tubulin heterodimers, with Rev bound on the inside. Rev has a strong inhibitory effect on aster formation in Xenopus egg extracts, demonstrating that it can interact with tubulin in the presence of normal levels of cellular constituents. These results suggest that Rev may interact with MTs to induce their destabilization, a proposition consistent with the previously described disruption of MTs after HIV-1 infection. C1 NICHD, Lab Integrat & Med Biophys, NIH, Bethesda, MD 20892 USA. NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. NIAMSD, Prot Express Lab, NIH, Bethesda, MD 20892 USA. NINDS, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. Wright State Univ, Dept Biol Sci, Dayton, OH 45435 USA. RP Sackett, DL (reprint author), NICHD, Lab Integrat & Med Biophys, NIH, Bldg 12A,Rm 2041, Bethesda, MD 20892 USA. FU NCRR NIH HHS [P41 RR001777, P41RR01777] NR 82 TC 34 Z9 34 U1 0 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP 349 EP 360 DI 10.1083/jcb.150.2.349 PG 12 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400018 PM 10908577 ER PT J AU Aoyagi, N Wassarman, DA AF Aoyagi, N Wassarman, DA TI Genes encoding Drosophila melanogaster RNA polymerase II general transcription factors: Diversity in TFIIA and TFIID components contributes to gene-specific transcriptional regulation SO JOURNAL OF CELL BIOLOGY LA English DT Article ID BINDING PROTEIN; TATA BOX; TAFS; ELEMENT; FAMILY; CORE C1 NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Wassarman, DA (reprint author), NICHHD, Cell Biol & Metab Branch, NIH, Bldg 18T,Rm 101, Bethesda, MD 20892 USA. NR 35 TC 48 Z9 49 U1 0 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP F45 EP F49 DI 10.1083/jcb.150.2.F45 PG 5 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400005 PM 10908585 ER PT J AU Brody, T Cravchik, A AF Brody, T Cravchik, A TI Drosophila melanogaster G protein-coupled receptors SO JOURNAL OF CELL BIOLOGY LA English DT Article ID MUSCARINIC ACETYLCHOLINE-RECEPTOR; DEVELOPMENTAL REGULATION; FUNCTIONAL EXPRESSION; OCTOPAMINE RECEPTOR; SEROTONIN RECEPTOR; CLONING; FAMILY; NEUROTRANSMITTER; LOCALIZATION; NEUROPEPTIDE C1 Celera Genom, Rockville, MD 20850 USA. NINDS, Neurogenet Unit, NIH, Bethesda, MD 20892 USA. RP Cravchik, A (reprint author), Celera Genom, 45 W Gude Dr, Rockville, MD 20850 USA. RI Marion-Poll, Frederic/D-8882-2011 OI Marion-Poll, Frederic/0000-0001-6824-0180 NR 40 TC 117 Z9 135 U1 0 U2 16 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP F83 EP F88 DI 10.1083/jcb.150.2.F83 PG 6 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400011 PM 10908591 ER PT J AU Fortini, ME Skupski, MP Boguski, MS Hariharan, IK AF Fortini, ME Skupski, MP Boguski, MS Hariharan, IK TI A survey of human disease gene counterparts in the Drosophila genome SO JOURNAL OF CELL BIOLOGY LA English DT Article ID P53; DEGENERATION; SUPPRESSION; INTERACTS; MUTATIONS; HOMOLOG; KINASE; MDM2 C1 Univ Penn, Sch Med, Dept Genet, Stellar Chance Labs, Philadelphia, PA 19104 USA. Celera Genom, Rockville, MD 20850 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. Massachusetts Gen Hosp, Ctr Canc, Charlestown, MA 02129 USA. RP Fortini, ME (reprint author), Univ Penn, Sch Med, Dept Genet, Stellar Chance Labs, Room 709C,422 Curie Blvd, Philadelphia, PA 19104 USA. RI Tang, Amy/L-3226-2016 OI Tang, Amy/0000-0002-5772-2878 FU NEI NIH HHS [R01EY11632]; NIA NIH HHS [R01 AG014583, R01AG14583]; NINDS NIH HHS [R01 NS036084, R01NS36084] NR 28 TC 103 Z9 110 U1 1 U2 20 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP F23 EP F29 DI 10.1083/jcb.150.2.F23 PG 7 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400002 PM 10908582 ER PT J AU Morrison, DK Murakami, MS Cleghon, V AF Morrison, DK Murakami, MS Cleghon, V TI Protein kinases and phosphatases in the Drosophila genome SO JOURNAL OF CELL BIOLOGY LA English DT Article ID TYROSINE KINASE; SIGNAL-TRANSDUCTION; FAMILY; IDENTIFICATION; ENCODES C1 NCI, Frederick Canc Res & Dev Ctr, Regulat Cell Growth Lab, Frederick, MD 21702 USA. RP Morrison, DK (reprint author), NCI, Frederick Canc Res & Dev Ctr, Regulat Cell Growth Lab, POB B, Frederick, MD 21702 USA. NR 29 TC 97 Z9 98 U1 1 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 24 PY 2000 VL 150 IS 2 BP F57 EP F62 DI 10.1083/jcb.150.2.F57 PG 6 WC Cell Biology SC Cell Biology GA 339DM UT WOS:000088462400007 PM 10908587 ER PT J AU Luke, BT AF Luke, BT TI Comparison of different data set screening methods for use in QSAR/QSPR generation studies SO JOURNAL OF MOLECULAR STRUCTURE-THEOCHEM LA English DT Article DE data set screening; quantitative structure/property relationship; evolutionary programming ID STRUCTURE-PROPERTY RELATIONSHIPS; GENETIC FUNCTION APPROXIMATION; DESCRIPTORS; PREDICTION; SHAPE AB In investigations aimed at generating accurate Quantitative Structure/Activity Relationships (QSAR) or Quantitative Structure/Property Relationships (QSPR), data sets are used that potentially contain a large number of descriptors for each compound. For trample, one of the data sets generated by Breneman and Rhem [J. Comput. Chem. 18 (1997) 182-197] contain 118 descriptors and the HPLC capacity factors [log(k')] in an ODS column for 22 compounds. One method of improving the search for good relationships is to prescreen the data set and, hopefully, remove redundant descriptors. This paper examines six different methods of prescreening a data set. Each method is used to generate multiple subsets of the data, either using different screening thresholds or producing sets with a given number of descriptors. Each set is then examined in two different ways. The first uses an Evolutionary Programming method described earlier to generate multiple relationships, and the second determines which relationships could be obtained from this reduced set of descriptors and ranks them relative to the top 250 relationships obtained from an All Possible Sets search of the full data set. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. RP NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Adv Biomed Comp Ctr, 430 Miller Dr, Frederick, MD 21702 USA. NR 22 TC 6 Z9 6 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-1280 J9 J MOL STRUC-THEOCHEM JI Theochem-J. Mol. Struct. PD JUL 24 PY 2000 VL 507 BP 229 EP 238 DI 10.1016/S0166-1280(99)00404-2 PG 10 WC Chemistry, Physical SC Chemistry GA 336MH UT WOS:000088305800024 ER PT J AU Nawab, SS Miller, CS Dale, JK Greenberg, BD Friedman, TC Chrousos, GP Straus, SE Rosenthal, NE AF Nawab, SS Miller, CS Dale, JK Greenberg, BD Friedman, TC Chrousos, GP Straus, SE Rosenthal, NE TI Self-reported sensitivity to chemical exposures in five clinical populations and healthy controls SO PSYCHIATRY RESEARCH LA English DT Article DE seasonal affective disorder; chronic fatigue syndrome; Addison's disease; obsessive-compulsive disorder; Cushing's syndrome; hypothalamic-pituitary-adrenal axis (HPA); multiple chemical sensitivity; chemical exposure ID CHRONIC FATIGUE SYNDROME; SEASONAL AFFECTIVE-DISORDER; PREVALENCE; DIAGNOSIS AB Two hundred and twenty-five subjects, including normal volunteers and patients with previously documented seasonal affective disorder (SAD), chronic fatigue syndrome (CFS), Cushing's syndrome, Addison's disease and obsessive-compulsive disorder (OCD), completed a self-rated inventory of reported sensitivity to various chemical exposures. Patients with CFS, Addison's disease and SAD self-reported more sensitivity to chemical exposures than normal controls. In addition, women reported more sensitivity than men. This report suggests that chemical sensitivity may be a relevant area to explore in certain medical and psychiatric populations. A possible relationship between reported chemical sensitivity and hypothalamic-pituitary-adrenal (HPA)-axis functioning is discussed. (C) Published by Elsevier Science Ireland Ltd. C1 NIMH, Sect Biol Rhythms, Bethesda, MD 20892 USA. Univ Texas, Hlth Sci Ctr, Dept Family Practice Environm & Occupat Med, San Antonio, TX 78284 USA. NIAID, Clin Invest Lab, Bethesda, MD 20892 USA. NIMH, Clin Sci Lab, Bethesda, MD 20892 USA. NICHHD, Dev Neurobiol Lab, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Cedars Sinai Med Ctr, Sch Med, Burns & Allen Res Inst,Dept Med,Div Endocrinol, Los Angeles, CA 90048 USA. NICHHD, Dev Endocrinol Branch, Bethesda, MD 20892 USA. RP Nawab, SS (reprint author), 10700 Stapleford Hall Dr, Potomac, MD 20854 USA. NR 28 TC 11 Z9 12 U1 1 U2 2 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0165-1781 J9 PSYCHIAT RES JI Psychiatry Res. PD JUL 24 PY 2000 VL 95 IS 1 BP 67 EP 74 DI 10.1016/S0165-1781(00)00148-7 PG 8 WC Psychiatry SC Psychiatry GA 341AP UT WOS:000088569100007 PM 10904124 ER PT J AU Anderson, DW Probst, FJ Belyantseva, IA Fridell, RA Beyer, L Martin, DM Wu, D Kachar, B Friedman, TB Raphael, Y Camper, SA AF Anderson, DW Probst, FJ Belyantseva, IA Fridell, RA Beyer, L Martin, DM Wu, D Kachar, B Friedman, TB Raphael, Y Camper, SA TI The motor and tail regions of myosin XV are critical for normal structure and function of auditory and vestibular hair cells SO HUMAN MOLECULAR GENETICS LA English DT Article ID UNCONVENTIONAL MYOSIN; DEAFNESS DFNB3; INNER-EAR; PROTEINS; DOMAIN; GENE; SHAKER-2; HOMOLOGY; MEMBRANE; ENCODES AB Recessive mutations in myosin 15, a class XV unconventional myosin, cause profound congenital deafness in humans and both deafness and vestibular dysfunction in mice homozygous for the shaker 2 and shaker 2(J) alleles, The shaker 2 allele is a previously described missense mutation of a highly conserved residue in the motor domain of myosin XV, The shaker 2(J) lesion, in contrast, is a 14.7 kb deletion that removes the last six exons from the 3'-terminus of the Myo 15 transcript, These exons encode a FERM (F, ezrin, radixin and moesin) domain that may interact with integral membrane proteins, Despite the deletion of six exons, Myo15 mRNA transcripts and protein are present in the post-natal day 1 shaker 2(J) inner ear, which suggests that the FERM domain is critical for the development of normal hearing and balance, Myo15 transcripts are first detectable at embryonic day 13.5 in wild-type mice, Myo15 transcripts in the mouse inner ear are restricted to the sensory epithelium of the developing cristae ampularis, macula utriculi and macula sacculi of the vestibular system as well as to the developing organ of Corti, Both the shaker 2 and shaker 2(J) alleles result in abnormally short hair cell stereocilia in the cochlear and vestibular systems, This suggests that Myo15 may be important for both the structure and function of these sensory epithelia. C1 Univ Michigan, Dept Human Genet, Ann Arbor, MI 48109 USA. Univ Michigan, Dept Otolaryngol, Ann Arbor, MI 48109 USA. Univ Michigan, Dept Pediat & Communicable Dis, Ann Arbor, MI 48109 USA. NIDCD, Lab Cellular Biol, Bethesda, MD 20892 USA. NIDCD, Mol Genet Lab, Rockville, MD 20850 USA. NIDCD, Cell Biol Lab, Rockville, MD 20850 USA. RP Camper, SA (reprint author), Univ Michigan, Dept Human Genet, 4301 MSRB 3,1150 W Med Ctr Dr, Ann Arbor, MI 48109 USA. OI Martin, Donna/0000-0002-8070-2007; Camper, Sally/0000-0001-8556-3379 FU NICHD NIH HHS [HD30428]; NIDCD NIH HHS [1 Z01 DC00048-02, DC016434] NR 39 TC 66 Z9 66 U1 0 U2 3 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 22 PY 2000 VL 9 IS 12 BP 1729 EP 1738 DI 10.1093/hmg/9.12.1729 PG 10 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 341LQ UT WOS:000088592200002 PM 10915760 ER PT J AU Lee, S Kozlov, S Hernandez, L Chamberlain, SJ Brannan, CI Stewart, CL Wevrick, R AF Lee, S Kozlov, S Hernandez, L Chamberlain, SJ Brannan, CI Stewart, CL Wevrick, R TI Expression and imprinting of MAGEL2 suggest a role in Prader-Willi syndrome and the homologous murine imprinting phenotype SO HUMAN MOLECULAR GENETICS LA English DT Article ID SYNDROME CRITICAL REGION; NECDIN GENE; MESSENGER-RNA; MOUSE; LETHALITY; PROTEIN; SNRPN; MODEL AB Prader-Willi syndrome (PWS) is caused by the loss of expression of imprinted genes in chromosome 15q11-q13. Affected individuals exhibit neonatal hypotonia, developmental delay and childhood-onset obesity. Necdin, a protein implicated in the terminal differentiation of neurons, is the only PWS candidate gene to reduce viability when disrupted in a mouse model, In this study, we have characterized MAGEL2 (also known as NDNL1), a gene with 51% amino acid sequence similarity to necdin and located 41 kb distal to NDN in the PWS deletion region. MAGEL2 is expressed predominantly in brain, the primary tissue affected in PWS and in several fetal tissues as shown by northern blot analysis. MAGEL2 is imprinted with monoallelic expression in control brain, and paternal-only expression in the central nervous system as demonstrated by its lack of expression in brain from a PWS-affected individual. The orthologous mouse gene (Magel2) is located within 150 kb of Ndn, is imprinted with paternal-only expression and is expressed predominantly in late developmental stages and adult brain as shown by northern blotting, RT-PCR and whole-mount RNA in situ hybridization, Magel2 distribution partially overlaps that of Ndn, with strong expression being detected in the central nervous system in midgestation mouse embryos by in situ hybridization, We hypothesize that, although loss of necdin expression may be important in the neonatal presentation of PWS, loss of MAGEL2 may be critical to abnormalities in brain development and dysmorphic features in individuals with PWS. C1 Univ Alberta, Dept Med Genet, Edmonton, AB T6G 2H7, Canada. NCI, Lab Canc Dev Biol, FCRDC, Frederick, MD 21702 USA. Univ Florida, Coll Med, Dept Mol Genet & Microbiol, Gainesville, FL 32610 USA. Univ Florida, Coll Med, Ctr Mammalian Genet, Gainesville, FL 32610 USA. Univ Florida, Coll Med, Inst Brain, Gainesville, FL 32610 USA. RP Wevrick, R (reprint author), Univ Alberta, Dept Med Genet, 842 Med Sci Bldg, Edmonton, AB T6G 2H7, Canada. OI Wevrick, Rachel/0000-0002-3343-5794 NR 21 TC 72 Z9 74 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 22 PY 2000 VL 9 IS 12 BP 1813 EP 1819 DI 10.1093/hmg/9.12.1813 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 341LQ UT WOS:000088592200012 PM 10915770 ER PT J AU Ai, YJ Zheng, Z O'Brien-Jenkins, A Bernard, DJ Wynshaw-Boris, T Ning, C Reynolds, R Segal, S Huang, K Stambolian, D AF Ai, YJ Zheng, Z O'Brien-Jenkins, A Bernard, DJ Wynshaw-Boris, T Ning, C Reynolds, R Segal, S Huang, K Stambolian, D TI A mouse model of galactose-induced cataracts SO HUMAN MOLECULAR GENETICS LA English DT Article ID GALACTOKINASE DEFICIENCY; ALDOSE REDUCTASE; GENE; MICE; LENS AB Galactokinase (GK; EC 2.7.1.6) is the first enzyme in the metabolism of galactose, In humans, GK deficiency results in congenital cataracts due to an accumulation of galactitol within the lens. In an attempt to make a galactosemic animal model, we cloned the mouse GK gene (Glk1) and disrupted it by gene targeting. As expected, galactose was very poorly metabolized in GK-deficient mice. In addition, both galactose and galactitol accumulated in tissues of GK-deficient mice. Surprisingly, the GK-deficient animals did not form cataracts even when fed a high galactose diet. However, the introduction of a human aldose reductase transgene into a GK-deficient background resulted in cataract formation within the first postnatal day. This mouse represents the first mouse model for congenital galactosemic cataract. C1 Univ Penn, Dept Ophthalmol, Philadelphia, PA 19104 USA. NHGRI, Genet Dis Branch, Bethesda, MD USA. NIH, Genet Dis Branch, Bethesda, MD USA. Childrens Hosp Philadelphia, Div Biochem Dev & Mol Dis, Philadelphia, PA 19104 USA. RP Stambolian, D (reprint author), Univ Penn, Dept Ophthalmol, Room 313 Stellar Chance,422 Curie Blvd, Philadelphia, PA 19104 USA. FU NEI NIH HHS [R01 EY09404]; NICHD NIH HHS [P01 HD29847] NR 22 TC 40 Z9 42 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 22 PY 2000 VL 9 IS 12 BP 1821 EP 1827 DI 10.1093/hmg/9.12.1821 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 341LQ UT WOS:000088592200013 PM 10915771 ER PT J AU Levy, D Kannel, WB AF Levy, D Kannel, WB TI Searching for answers to ethnic disparities in cardiovascular risk SO LANCET LA English DT Editorial Material ID CORONARY HEART-DISEASE C1 NHLBI, Framingham Heart Study, Framingham, MA 01702 USA. Boston Univ, Sch Med, Framingham, MA USA. RP Levy, D (reprint author), NHLBI, Framingham Heart Study, Framingham, MA 01702 USA. NR 9 TC 20 Z9 20 U1 0 U2 0 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUL 22 PY 2000 VL 356 IS 9226 BP 266 EP 267 DI 10.1016/S0140-6736(00)02496-X PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 336MJ UT WOS:000088305900002 PM 11071176 ER PT J AU Sher, L AF Sher, L TI Pharmacogenetics and antipsychotic drugs SO LANCET LA English DT Letter ID CLOZAPINE C1 NIMH, Bethesda, MD 20892 USA. RP Sher, L (reprint author), NIMH, Bethesda, MD 20892 USA. NR 5 TC 0 Z9 0 U1 1 U2 1 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUL 22 PY 2000 VL 356 IS 9226 BP 342 EP 342 DI 10.1016/S0140-6736(05)73627-8 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 336MJ UT WOS:000088305900054 PM 11071218 ER PT J AU Wosikowski, K Silverman, JA Bishop, P Mendelsohn, J Bates, SE AF Wosikowski, K Silverman, JA Bishop, P Mendelsohn, J Bates, SE TI Reduced growth rate accompanied by aberrant epidermal growth factor signaling in drug resistant human breast cancer cells SO BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH LA English DT Article ID MAMMARY EPITHELIAL-CELLS; RECEPTOR MONOCLONAL-ANTIBODIES; PROTEIN-KINASE-B; ADRIAMYCIN-RESISTANT; C-ERBB-2 EXPRESSION; ADJUVANT THERAPY; GENE; DOXORUBICIN; LINE; EGF AB We examined transforming growth factor (TGF) alpha, epidermd growth factor (EGF) and EGF receptor (EGFR) expression and signaling in three drug resistant MCF-7 human breast cancer sublines and asked whether these pathways contribute to the drug resistance phenotype. In the resistant sublines, upregulation of both TGF alpha and EGFR mRNA was observed. In an apparent contrast with upregulated growth factor and receptor gene expression, the drug resistant sublines displayed a reduced growth rate. Defects in the EGFR signaling pathway cascade were found in all examined drug resistant sublines, including altered EGF-induced Shc, Raf-1, or mitogen-activated protein kinase phosphorylation. Induction of c-fos mRNA expression by EGF was impaired in the sublines compared to parental MCF-7 cells. In contrast, the induction of the stress-activated protein kinase activity was similar in both parental and drug resistant cells. Evaluating the link between the reduced growth rate and drug resistance, serum starvation experiments were performed. These studies demonstrated that a reduced proliferative activity resulted in a marked reduction in sensitivity to cytotoxic agents in the parental MCF-7 cells. We propose that the altered EGFR levels frequently observed in drug resistant breast cancer cells are associated with perturbations in the signaling pathway that mediate a reduced proliferative rate and thereby contribute to drug resistance. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Div Clin Sci, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Div Basic Sci, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. RP Bates, SE (reprint author), NCI, Div Clin Sci, Med Branch, NIH, Bldg 10,Room 12N226,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 56 TC 14 Z9 14 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4889 J9 BBA-MOL CELL RES JI Biochim. Biophys. Acta-Mol. Cell Res. PD JUL 21 PY 2000 VL 1497 IS 2 BP 215 EP 226 DI 10.1016/S0167-4889(00)00062-8 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 338VW UT WOS:000088444800005 PM 10903426 ER PT J AU Bochar, DA Wang, L Beniya, H Kinev, A Xue, YT Lane, WS Wang, WD Kashanchi, F Shiekhattar, R AF Bochar, DA Wang, L Beniya, H Kinev, A Xue, YT Lane, WS Wang, WD Kashanchi, F Shiekhattar, R TI BRCA1 is associated with a human SWI/SNF-related complex: Linking chromatin remodeling to breast cancer SO CELL LA English DT Article ID POLYMERASE-II HOLOENZYME; TRANSCRIPTIONAL ACTIVATION; MEIOTIC CELLS; DNA-DAMAGE; EXPRESSION; PROTEIN; REPAIR; DOMAIN; GROWTH; BINDS AB Germline mutations in the tumor suppressor gene, BRCA1, predispose individuals to breast and ovarian cancers. Using a combination of affinity- and conventional chromatographic techniques, we have isolated a predominant form of a multiprotein BRCA1-containing complex from human cells displaying chromatin-remodeling activity. Mass spectrometric sequencing of components of this complex indicated that BRCA1 is associated with a SWI/SNF-related complex. We show that BRCA1 can directly interact with the BRG1 subunit of the SWI/SNF complex. Moreover, p53-mediated stimulation of transcription by BRCA1 was completely abrogated by either a dominant-negative mutant of BRG1 or the cancer-causing deletion in exon 11 of BRCA1. These findings reveal a direct function for BRCA1 in transcriptional control through modulation of chromatin structure. C1 Wistar Inst, Philadelphia, PA 19104 USA. Harvard Univ, Harvard Microchem Facil, Cambridge, MA 02138 USA. NIA, Genet Lab, NIH, Gerontol Res Ctr, Baltimore, MD 21224 USA. Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Biochem & Mol Biol, Newark, NJ 07103 USA. RP Shiekhattar, R (reprint author), Wistar Inst, 3601 Spruce St, Philadelphia, PA 19104 USA. RI Bochar, Daniel/A-9224-2009 NR 36 TC 374 Z9 388 U1 3 U2 18 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD JUL 21 PY 2000 VL 102 IS 2 BP 257 EP 265 DI 10.1016/S0092-8674(00)00030-1 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 337NA UT WOS:000088366800015 PM 10943845 ER PT J AU Kepplinger, KJF Kahr, H Forstner, G Sonnleitner, M Schindler, H Schmidt, T Groschner, K Soldatov, NM Romanin, C AF Kepplinger, KJF Kahr, H Forstner, G Sonnleitner, M Schindler, H Schmidt, T Groschner, K Soldatov, NM Romanin, C TI A sequence in the carboxy-terminus of the alpha(1C) subunit important for targeting, conductance and open probability of L-type Ca2+ channels SO FEBS LETTERS LA English DT Article DE class C-type Ca2+ channel; carboxyl tail; targeting; conductance; inactivation; fluorescence microscopy; GFP-labeled alpha(1C) subunit ID CALCIUM CHANNELS; BETA-SUBUNIT; CREB PHOSPHORYLATION; INACTIVATION; CALMODULIN; INHIBITION; MODULATION; DIVERSITY; SUPPORTS; NUCLEUS AB The role of the 80-amino acid motif 1572-1651 in the C-terminal tail of alpha(1C) Ca2+ channel subunits was studied by comparing properties of the conventional alpha(1C,77) channel expressed in HEK-tsA201 cells to three isoforms carrying alterations in this motif, Replacement of amino acids 1572-1651 in alpha(1C,77) with 81 non-identical residues leading to alc,ss impaired membrane targeting and cluster formation of the channel. Similar to alpha(1C,86), substitution of its 1572-1598 (alpha(1C,77L)) or 1595-1652 (alpha(1C,77K)) segments into the alpha(1C,77) channel yielded single-channel Ba2+ currents with increased inactivation, reduced open probability and unitary conductance, when compared to the alc,77 channel. Thus, the C-terminal sequence 1572-1651 of the ale subunit is important for membrane targeting, permeation and open probability of L-type Ca2+ channels. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 Univ Linz, Inst Biophys, A-4040 Linz, Austria. Leiden Univ, Dept Biophys, Leiden, Netherlands. Graz Univ, Inst Pharmacol & Toxicol, A-8010 Graz, Austria. NIA, NIH, Baltimore, MD 21224 USA. RP Romanin, C (reprint author), Univ Linz, Inst Biophys, Altenbergerstr 69, A-4040 Linz, Austria. RI Schmidt, Thomas/B-6296-2009; Groschner, Klaus/A-2550-2010; Romanin, Christoph/D-5399-2009 OI Schmidt, Thomas/0000-0002-0045-1851; Romanin, Christoph/0000-0003-3756-4136 NR 33 TC 24 Z9 24 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 21 PY 2000 VL 477 IS 3 BP 161 EP 169 DI 10.1016/S0014-5793(00)01791-9 PG 9 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 338PX UT WOS:000088429400003 PM 10908714 ER PT J AU Lee, CH Marekov, LN Kim, SY Brahim, JS Park, MH Steinert, PM AF Lee, CH Marekov, LN Kim, SY Brahim, JS Park, MH Steinert, PM TI Small proline-rich protein 1 is the major component of the cell envelope of normal human oral keratinocytes SO FEBS LETTERS LA English DT Article DE oral keratinocyte; barrier function; transglutaminase; cell envelope; small proline-rich protein ID TRANSGLUTAMINASE CROSS-LINKING; STRATIFIED SQUAMOUS EPITHELIA; EPIDERMAL-KERATINOCYTES; DIFFERENTIAL EXPRESSION; FAMILY; LORICRIN; INVOLUCRIN; BARRIER; ENZYME AB Oral keratinocytes of buccal and gingival tissues undergo a terminal differentiation program to form a protective epithelial barrier as non-keratinized or parakeratinized stratified cells, We have examined the protein composition of cell envelopes (CEs) from normal human buccal and gingival tissues as well as keratinocytes from normal human gingival cells grown in culture. Biochemical and sequencing analyses reveal that the CEs contain 60-70% small proline-rich protein 1a/b (SPR1a/b), together with smaller amounts of involucrin, annexin I and several other known CE proteins. The data imply a specialized role for SPR1 proteins in the unique barrier function requirements of oral epithelia, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V, All rights reserved. C1 NIAMS, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. NIDCR, Clin Res Core, NIH, Bethesda, MD 20892 USA. RP Steinert, PM (reprint author), NIAMS, Skin Biol Lab, NIH, Bldg 6,Room 425, Bethesda, MD 20892 USA. NR 29 TC 20 Z9 20 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 21 PY 2000 VL 477 IS 3 BP 268 EP 272 DI 10.1016/S0014-5793(00)01806-8 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 338PX UT WOS:000088429400022 PM 10908733 ER PT J AU Lau, OD Courtney, AD Vassilev, A Marzilli, LA Cotter, RJ Nakatani, Y Cole, PA AF Lau, OD Courtney, AD Vassilev, A Marzilli, LA Cotter, RJ Nakatani, Y Cole, PA TI p300/CBP-associated factor histone acetyltransferase processing of a peptide substrate SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SEROTONIN N-ACETYLTRANSFERASE; MELATONIN RHYTHM ENZYME; CATALYTIC MECHANISM; COACTIVATORS P300; CRYSTAL-STRUCTURE; COENZYME-A; DNA-DAMAGE; ACETYLATION; PCAF; BINDING AB p300/CBP-associated factor (PCAF) is a histone acetyltransferase that; plays an important role in the remodeling of chromatin and the regulation of gene expression. It has been shown to catalyze preferentially acetylation of the epsilon-amino group of lysine 14 in histone H3, In this study, the kinetic mechanism of PCAF was evaluated with a 20-amino acid peptide substrate derived from the amino terminus of histone H3 (H3-20) and recombinant bacterially expressed PCAF catalytic domain (PCAF(cat)). The enzymologic behavior of full-length PCAF and PCAF(cat) were shown to be similar. PCAF-catalyzed acetylation of the substrate H3-20 was shown to be specific for Lys-14, analogous to its behavior with the full-length histone H3 protein. Two-substrate kinetic analysis displayed an intersecting line pattern, consistent with a ternary complex mechanism for PCAF, The dead-end inhibitor analog desulfo-CoA was competitive versus acetyl-CoA and noncompetitive versus H3-20, The dead-end analog inhibitor H3-20 K14A was competitive versus H3-20 and uncompetitive versus acetylCoA The potent bisubstrate analog inhibitor H3-CoA-20 was competitive versus acetyl-CoA and noncompetitive versus H3-20. Taken together, these inhibition patterns support an ordered BiBi kinetic mechanism for PCAF in which acetyl-CoA binding precedes H3-20 binding. Viscosity experiments suggest that diffusional release of product is not rate-determining for PCAF catalysis, These results provide a mechanistic framework for understanding the detailed catalytic behavior of an important subset of the histone acetyltransferases and have significant implications for molecular regulation of and inhibitor design for these enzymes. C1 Johns Hopkins Univ, Sch Med, Dept Pharmacol & Mol Sci, Baltimore, MD 21205 USA. NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. Dana Farber Canc Inst, Boston, MA 02115 USA. RP Cole, PA (reprint author), Johns Hopkins Univ, Sch Med, Dept Pharmacol & Mol Sci, Rm 316,Hunterian Bldg,725 N Wolfe St, Baltimore, MD 21205 USA. NR 35 TC 82 Z9 82 U1 2 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 21953 EP 21959 DI 10.1074/jbc.M003219200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800029 PM 10777508 ER PT J AU Sukhodolets, MV Jin, DJ AF Sukhodolets, MV Jin, DJ TI Interaction between RNA polymerase and RapA, a bacterial homolog of the SWI/SNF protein family SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TRANSCRIPT CLEAVAGE FACTORS; ESCHERICHIA-COLI; GENE-EXPRESSION; MUTAGENESIS; GREA; RESISTANCE; HELICASES; LOCATION; BINDING; SUBUNIT AB Recently, we identified a novel Escherichia coli RNA polymerase (RNAP)-associated protein, an ATPase, called RapA (Sukhodolets, M, V., and Jin, D. J. (1998) J. Biol. Chem. 273, 7018-7023), RapA is a bacterial homolog of SWI2/SNF2. We showed that RapA forms a stable complex with RNAP holoenzyme and that binding to RNAP holoenzyme stimulates the ATPase activity of RapA. We have further analyzed the interactions between purified RapA and the two forms of RNAP: core RNAP and RNAP holoenzyme, We found that RapA interacts with either form of RNAP, However, RapA exhibits higher affinity for core RNAP than for RNAP holoenzyme. Chemical cross-linking of the RNAP-RapA complex indicated that the RapA-binding sites are located at the interface between the alpha and beta' subunits of RNAP, Contrary to previously reported results (Muzzin, O,, Campbell, E., A, Xia, L., Severinova, E., Darst, S. A, and Severinov, K, (1998) J. Biol. Chem, 273, 15157-15161), our in vivo analysis of a rapA null mutant suggested that RapA is not likely to be directly involved in DNA repair. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Jin, DJ (reprint author), NCI, Mol Biol Lab, NIH, Bldg 37,Rm 2E14,9000 Rockville Pike, Bethesda, MD 20892 USA. EM djjin@helix.nih.gov NR 32 TC 16 Z9 17 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 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22090 EP 22097 DI 10.1074/jbc.M000056200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800049 PM 10801781 ER PT J AU Watanabe, K Fujii, H Takahashi, T Kodama, M Aizawa, Y Ohta, Y Ono, T Hasegawa, G Naito, M Nakajima, T Kamijo, Y Gonzalez, FJ Aoyama, T AF Watanabe, K Fujii, H Takahashi, T Kodama, M Aizawa, Y Ohta, Y Ono, T Hasegawa, G Naito, M Nakajima, T Kamijo, Y Gonzalez, FJ Aoyama, T TI Constitutive regulation of cardiac fatty acid metabolism through peroxisome proliferator-activated receptor alpha associated with age-dependent cardiac toxicity SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ACYL-COA DEHYDROGENASE; COENZYME-A DEHYDROGENASE; ENZYME GENE-EXPRESSION; PPAR-ALPHA; BETA-OXIDATION; PENTADECANOIC ACID; MOLECULAR-CLONING; KAPPA-B; RAT; MITOCHONDRIAL AB The peroxisome proliferator-activated receptor alpha (PPAR alpha) is a member of the nuclear receptor superfamily and mediates the biological effects of peroxisome proliferators, To determine the physiological role of PPAR alpha in cardiac fatty acid metabolism, we examined the regulation of expression of cardiac fatty acid-metabolizing proteins using PPAR alpha-null mice. The capacity for constitutive myocardial beta-oxidation of the medium and long chain fatty acids, octanoic acid and palmitic acid, was markedly reduced in the PPAR alpha-null mice as compared with the wild-type mice, indicating that mitochondrial fatty acid catabolism is impaired in the absence of PPAR alpha. In contrast, constitutive beta-oxidation of the very long chain fatty acid, lignoceric acid, did not differ between the mice, suggesting that the constitutive expression of enzymes involved in peroxisomal beta-oxidation is independent of PPAR alpha. Indeed, PPAR alpha-null mice had normal levels of the peroxisomal beta-oxidation enzymes except the D-type bifunctional protein. At least seven mitochondrial fatty acid-metabolizing enzymes were expressed at much lower levels in the PPAR alpha-null mice, whereas other fatty acid-metabolizing enzymes were present at similar or slightly lower levels in the PPAR alpha-null, as compared with wild-type mice. Additionally, lower constitutive mRNA expression levels of fatty acid transporters were found in the PPAR alpha-null mice, suggesting a role for PPAR alpha in fatty acid transport and catabolism. Indeed, in fatty acid metabolism experiments in vivo, myocardial uptake of iodophenyl 9-methylpentadecanoic acid and its conversion to 3-methyl-nonanoic acid were reduced in the PPAR alpha-null mice. Interestingly, a decreased ATP concentration after exposure to stress, abnormal cristae of the mitochondria, abnormal caveolae, and fibrosis were observed only in the myocardium of the PPAR alpha-null mice. These cardiac abnormalities appeared to proceed in an age-dependent manner. Taken together, the results presented here indicate that PPAR alpha controls constitutive fatty acid oxidation, thus establishing a role for the receptor in cardiac fatty acid homeostasis, Furthermore, altered expression of fatty acid-metabolizing proteins seems to lead to myocardial damage and fibrosis, as inflammation and abnormal cell growth control can cause these conditions. C1 Niigata Coll Pharm, Dept Clin Pharmacol, Niigata 9502081, Japan. Niigata Univ, Sch Med, Dept Biochem 2, Niigata 9518510, Japan. Niigata Univ, Sch Med, Radioisotope Ctr, Niigata 9518510, Japan. Niigata Univ, Sch Med, Dept Med 1, Niigata 9518510, Japan. Niigata Univ, Sch Med, Dept Pathol 2, Niigata 9518510, Japan. Shinshu Univ, Sch Med, Dept Hyg & Med Genet, Nagano 3908621, Japan. Shinshu Univ, Sch Med, Dept Internal Med 2, Nagano 3908621, Japan. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Shinshu Univ, Sch Med, Dept Aging Biochem, Matsumoto, Nagano 3908621, Japan. RP Watanabe, K (reprint author), Niigata Coll Pharm, Dept Clin Pharmacol, Kamisin Ei Cho, Niigata 9502081, Japan. NR 45 TC 195 Z9 208 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22293 EP 22299 DI 10.1074/jbc.M000248200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800074 PM 10801788 ER PT J AU Wang, W Soto, H Oldham, ER Buchanan, ME Homey, B Catron, D Jenkins, N Copeland, NG Gilbert, DJ Nguyen, N Abrams, J Kershenovich, D Smith, K McClanahan, T Vicari, AP Zlotnik, A AF Wang, W Soto, H Oldham, ER Buchanan, ME Homey, B Catron, D Jenkins, N Copeland, NG Gilbert, DJ Nguyen, N Abrams, J Kershenovich, D Smith, K McClanahan, T Vicari, AP Zlotnik, A TI Identification of a novel chemokine (CCL28), which binds CCR10 (GPR2) SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CXC CHEMOKINE; PROTEIN; CELLS; LYMPHOCYTES; LYMPHOTACTIN; EXPRESSION; TECK AB We report the identification and characterization of a novel CC chemokine designated CCL28 and its receptor CCR10, known previously as orphan G-protein-coupled receptor GPR2. Human and mouse CCL28 share 83% identity at the amino acid and 76% at the nucleic acid levels. We also identified the mouse homologues of CCL28 and of CCR10, which map to mouse chromosomes 13 and 11, respectively, CCL28 is expressed in a variety of human and mouse tissues, and it appears to be predominantly produced by epithelial cells. Both human and mouse CCL28 induce calcium mobilization in human and mouse CCL28 expressing transfectants. CCL28 desensitized the calcium mobilization induced in CCR10 transfectants by CCL27, indicating that these chemokines share this new chemokine receptor. In vitro, recombinant human CCL28 displays chemotactic activity for resting CD4 or CD8 T cells. C1 DNAX Res Inst Mol & Cellular Biol Inc, Dept Immunobiol, Palo Alto, CA 94304 USA. Schering Plough Lab Immunol Res, F-69572 Dardilly, France. NCI, Frederick Canc Res & Dev Ctr, Mammalian Genet Lab, Frederick, MD 21702 USA. Inst Nacl Nutr Salvador Zubiran, Dept Gastroenterol, Mexico City 14000, DF, Mexico. RP Zlotnik, A (reprint author), DNAX Res Inst Mol & Cellular Biol Inc, Dept Immunobiol, Palo Alto, CA 94304 USA. RI Zlotnik, Albert/C-3791-2011 NR 25 TC 169 Z9 187 U1 0 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22313 EP 22323 DI 10.1074/jbc.M001461200 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800077 PM 10781587 ER PT J AU Brooks, PJ Wise, DS Berry, DA Kosmoski, JV Smerdon, MJ Somers, RL Mackie, H Spoonde, AY Ackerman, EJ Coleman, K Tarone, RE Robbins, JH AF Brooks, PJ Wise, DS Berry, DA Kosmoski, JV Smerdon, MJ Somers, RL Mackie, H Spoonde, AY Ackerman, EJ Coleman, K Tarone, RE Robbins, JH TI The oxidative DNA lesion 8,5 '-(S)-cyclo-2 '-deoxyadenosine is repaired by the nucleotide excision repair pathway and blocks gene expression in mammalian cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PIGMENTOSUM COMPLEMENTATION GROUP; RADICAL-INDUCED FORMATION; XERODERMA-PIGMENTOSUM; 6-4 PHOTOPRODUCTS; PYRIMIDINE DIMER; RNA-POLYMERASE; FREE-EXTRACTS; PLASMID DNA; BASE DAMAGE; GROUP-F AB Xeroderma pigmentosum (XP) patients with inherited defects in nucleotide excision repair (NER) are unable to excise from their DNA bulky photoproducts induced by UV radiation and therefore develop accelerated actinic damage, including cancer, on sun-exposed tissue. Some XP patients also develop a characteristic neurodegeneration believed to result from their inability to repair neuronal DNA damaged by endogenous metabolites since the harmful UV radiation in sunlight does not reach neurons. Free radicals, which are abundant in neurons, induce DNA lesions that, if unrepaired, might cause the XP neurodegeneration. Searching for such a lesion, we developed a synthesis for 8,5'-(S)-cyclo-2'-deoxyadenosine (cyclo-dA), a free radical-induced bulky lesion, and incorporated it into DNA to test its repair in mammalian cell extracts and living cells. Using extracts of normal and mutant Chinese hamster ovary (CHO) cells to test for NER and adult rat brain extracts to test for base excision repair, we found that cyclo-dA is re paired by NER and not by base excision repair. We measured host cell reactivation, which reflects a cell's capacity for NER, by transfecting CHO and XP cells with DNA constructs containing a single cyclo-dA or a cyclobutane thymine dimer at a specific site on the transcribed strand of a luciferase reporter gene. We found that, like the cyclobutane thymine dimer, cyclo-dA is a strong block to gene expression in CHO and human cells. Cyclo-dA was repaired extremely poorly in NER-deficient CHO cells and in cells from patients in XP complementation group A with neurodegeneration. Based on these findings, we propose that cyclo-dA is a candidate for an endogenous DNA lesion that might contribute to neurodegeneration in XP. C1 NIAAA, Neurogenet Lab, NIH, Bethesda, MD 20892 USA. Berry Associates Inc, Dexter, MI 48130 USA. Washington State Univ, Dept Biochem & Biophys, Pullman, WA 99164 USA. Glen Res Corp, Sterling, VA 20164 USA. Pacific NW Natl Lab, Dept Energy, Richland, WA 99352 USA. NCI, Dermatol Branch, NIH, Bethesda, MD 20892 USA. NCI, Biostat Branch, NIH, Bethesda, MD 20892 USA. RP Brooks, PJ (reprint author), NIAAA, Neurogenet Lab, NIH, 12420 Parklawn Dr, Bethesda, MD 20892 USA. FU NIEHS NIH HHS [ES02614, ESO4106] NR 49 TC 179 Z9 182 U1 0 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22355 EP 22362 DI 10.1074/jbc.M002259200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800082 PM 10801836 ER PT J AU Zhang, P Wang, YZ Kagan, E Bonner, JC AF Zhang, P Wang, YZ Kagan, E Bonner, JC TI Peroxynitrite targets the epidermal growth factor receptor, Raf-1, and MEK independently to activate MAPK SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HYDROGEN-PEROXIDE; PROTEIN-KINASE; NITRIC-OXIDE; SIGNAL-TRANSDUCTION; PULMONARY FIBROSIS; INDUCED APOPTOSIS; IN-VITRO; CELLS; EXPRESSION; H2O2 AB Activation of ERK-1 and -2 by H2O2 in a variety of cell types requires epidermal growth factor receptor (EGFR) phosphorylation, In this study, we investigated the activation of ERK by ONOO- in cultured rat lung myofibroblasts. Western blot analysis using anti-phos-pho-ERK antibodies along with an ERK kinase assay using the phosphorylated heat- and acid-stable protein (PHAS-1) substrate demonstrated that ERK activation peaked within 15 min after ONOO- treatment and was maximally activated with 100 mu M ONOO-. Activation of ERK by ONOO- and H2O2 was blocked by the antioxidant N-acetyl-L-cysteine. Catalase blocked ERK activation by H2O2, but not by ONOO-, demonstrating that the effect of ONOO- was not due to the generation of H2O2, Both H2O2 and ONOO- induced phosphorylation of EGFR in Western blot experiments using an anti-phos-pho-EGFR antibody. However, the EGFR tyrosine kinase inhibitor AG1478 abolished ERK activation by H2O2, but not by ONOO-. Both H2O2 and ONOO- activated Raf-1, However, the Raf inhibitor forskolin blocked ERK activation by H2O2, but not by ONOO-. The MEK inhibitor PD98059 inhibited ERK activation by both H2O2 and ONOO-. Moreover, ONOO- or H2O2 caused a cytotoxic response of myofibroblasts that was prevented by preincubation with PD98059, In a cell-free kinase assay, ONOO- (but not H2O2) induced autophosphorylation and nitration of a glutathione S-transferase-MEK-1 fusion protein. Collectively, these data indicate that ONOO- activates EGFR and Raf-1, but these signaling intermediates are not required for ONOO--induced ERK activation. However, MEK-1 activation is required for ONOO--induced ERK activation in myofibroblasts, In contrast, H2O2-induced ERK activation is dependent on EGFR activation, which then leads to downstream Raf-1 and MEK-1 activation. C1 NIEHS, Pulm Pathobiol Lab, NIH, Res Triangle Pk, NC 27709 USA. Uniformed Serv Univ Hlth Sci, Dept Pathol, Bethesda, MD 20814 USA. RP Bonner, JC (reprint author), NIEHS, Pulm Pathobiol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. FU NHLBI NIH HHS [HL-54196] NR 37 TC 105 Z9 110 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22479 EP 22486 DI 10.1074/jbc.M910425199 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800097 PM 10801894 ER PT J AU Hagemann, D Kuschel, M Kuramochi, T Zhu, WZ Cheng, HP Xiao, RP AF Hagemann, D Kuschel, M Kuramochi, T Zhu, WZ Cheng, HP Xiao, RP TI Frequency-encoding Thr(17) phospholamban phosphorylation is independent of Ser(16) phosphorylation in cardiac myocytes SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BETA-ADRENERGIC STIMULATION; PROTEIN-KINASE; HEART-CELLS; INTACT HEART; MECHANISMS; SITES; CONTRACTILITY; RELAXATION; VENTRICLES; CANINE AB Both Ser(16) and Thr(17) Of phospholamban (PLB) are phosphorylated, respectively, by cAMP-dependent protein kinase (PKA) and Ca2+/calmodulin-dependent protein kinase II (CaMKII), PLB phosphorylation relieves cardiac sarcoplasmic reticulum Ca2+ pump from inhibition by PLB. Previous studies have suggested that phosphorylation of Ser(16) by PKA is a prerequisite for Thr(17) phosphorylation by CaMKII and is essential to the relaxant effect of beta-adrenergic stimulation. To determine the role of Thr(17) PLB phosphorylation, we investigated the dual-site phosphorylation of PLB in isolated adult rat cardiac myocytes in response to beta(1)-adrenergic stimulation or electrical field stimulation (0.1-3 Hz) or both. A beta(1)-adrenergic agonist, norepinephrine (10(-9)-10(-6) M), in the presence of an alpha(1)-adrenergic antagonist, prazosin (10(-6) hr), selectively increases the PKA-dependent phos- phorylation of PLB at Ser(16) in quiescent myocytes, In contrast, electrical pacing induces an opposite phosphorylation pattern, selectively enhancing the CaMKII-mediated Thr(17) PLB phosphorylation in a frequency-dependent manner. When combined, electric stimulation (2 Hz) and beta(1)-adrenergic stimulation lead to dual phosphorylation of PLB and exert a synergistic effect on phosphorylation of Thr(17) but not Ser(16) Frequency-dependent Thr(17) phosphorylation is closely correlated with a decrease in 50% relaxation time (t(50)) of cell contraction, which is independent of, but additive to, the relaxant effect of Ser(16) phosphorylation, resulting in hastened contractile relaxation at high stimulation frequencies. Thus, we conclude that in intact cardiac myocytes, phosphorylation of PLB at Thr(17) occurs in the absence of prior Ser(16) phosphorylation, and that frequencydependent Thr(17) PLB phosphorylation may provide an intrinsic mechanism for cardiac myocytes to adapt to a sudden change of heart rate. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Xiao, RP (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 31 TC 106 Z9 107 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22532 EP 22536 DI 10.1074/jbc.C000253200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800104 PM 10825152 ER PT J AU Wang, YZ Zhang, P Rice, AB Bonner, JC AF Wang, YZ Zhang, P Rice, AB Bonner, JC TI Regulation of interleukin-1 beta-induced platelet-derived growth factor receptor-alpha expression in rat pulmonary myofibroblasts by p38 mitogen-activated protein kinase SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID MESSENGER-RNA DEGRADATION; TUMOR-NECROSIS-FACTOR; AU-RICH ELEMENTS; FACTOR-KAPPA-B; MAP KINASE; SIGNAL-TRANSDUCTION; PDGF RECEPTOR; 3'-UNTRANSLATED REGION; CELLULAR STRESSES; GENE-EXPRESSION AB The potential role of p38 mitogen-activated protein (MAP) kinase in platelet-derived growth factor receptor-alpha (PDGF-R alpha) gene expression was investigated using cultured rat pulmonary myofibroblasts. p38 MAP kinase was constitutively expressed in myofibroblasts and activated by interleukin (IL)-1 beta. A pyridinylimidazole compound, SB203580, completely inhibited the ability of p38 MAP kinase activity to phosphorylate PHAS-1 substrate. SB203580 inhibited IL-1 beta-induced up-regulation of PDGF-R alpha mRNA and protein in a concentration-dependent manner. Other kinase inhibitors, including the mitogen-activated protein kinase/extracellular signal-regulated kinase inhibitor PD98059, did not block upregulation of PDGF-R alpha. The IL-1 beta-induced increase in the number of I-125 PDGF-AA-binding sites at the cell surface was reduced >70% by pretreatment with SB203580. Accordingly, an enhancement of PDGF-AA-stimulated DNA synthesis following IL-1 beta pretreatment was blocked > 70% by SB203580. SB203580 did not affect IL-1 beta-induced ERK activation, yet enhanced IL-1 beta-induced JNK activation approximately 2-fold. Treatment of cells with SB203580 after inhibition of transcription by actinomycin D decreased the half-life of IL-1 beta-induced PDGF-R alpha mRNA from >4 to similar to 1.5 h. Moreover, pretreatment of cells with cycloheximide blocked induction of PDGF-R alpha mRNA by IL-1 beta, suggesting that de novo protein synthesis was required for PDGF-R alpha mRNA stabilization. These data indicate that p38 MAP kinase regulates PDGF-R alpha expression at the translational level by signaling the synthesis of an mRNA-stabilizing protein. C1 NIEHS, Pulm Pathobiol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Bonner, JC (reprint author), NIEHS, Pulm Pathobiol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 50 TC 35 Z9 35 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22550 EP 22557 DI 10.1074/jbc.M909785199 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800107 PM 10807932 ER PT J AU Rajapandi, T Greene, LE Eisenberg, E AF Rajapandi, T Greene, LE Eisenberg, E TI The molecular chaperones Hsp90 and Hsc70 are both necessary and sufficient to activate hormone binding by glucocorticoid receptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HEAT-SHOCK-PROTEIN; FUNCTIONAL HETEROCOMPLEX; RETICULOCYTE LYSATE; HSP70; MACHINERY; COMPLEX; FOLDOSOME; RECONSTITUTION; SYSTEM; DNAJ AB Glucocorticoid receptors must be complexed with Hsp90 in order to bind steroids, and it has been reported that at least three other proteins, Hop, Hsc70, and a J-domain protein (either Hsp40 or Ydj1), are required for formation of active Hsp90-steroid receptor complex. In the present study, we reinvestigated activation of stripped steroid receptors isolated from either L cells or WCL2 cells. Surprisingly, we found, using highly purified proteins, that only Hsp90 and Hsc70 are required for the activation of glucocorticoid receptors in the presence of steroids; In the absence of steroids, either p23 or molybdate are also required as reported previously. Addition of Hop or Ydj1 had no affect on the rate or magnitude of the activation of the stripped receptors, and quantitative Western blots confirmed that neither Hop or Hsp40 were present in our protein preparations or in the stripped receptors. Furthermore, a truncated recombinant Hsp70 that does not bind Hop or Hsp40 was as effective as wild-type Hsp70 in activating stripped receptor. Since Hsc70 does not bind directly to Hsp90 but both proteins bind to Hop, it has been suggested that Hop acts as a bridge between Hsp90 and Hsp70. However, we found that after Hsc70 or Hsp90 bind directly to the stripped receptors, they are fully reactivated by Hsp90 or Hsc70, respectively. We, therefore, conclude that Hsp90 and Hsc70 bind independently to stripped glucocorticoid receptors and alone are sufficient to activate them to bind steroids. C1 NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Eisenberg, E (reprint author), NHLBI, Cell Biol Lab, NIH, 9000 Rockville Pike,3 Ctr Dr,MSC 0301,Bldg 2,Room, Bethesda, MD 20892 USA. NR 26 TC 70 Z9 71 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 21 PY 2000 VL 275 IS 29 BP 22597 EP 22604 DI 10.1074/jbc.M002035200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337LY UT WOS:000088363800114 PM 10781595 ER PT J AU Shibusawa, Y Yanagida, A Ito, A Ichihashi, K Shindo, H Ito, Y AF Shibusawa, Y Yanagida, A Ito, A Ichihashi, K Shindo, H Ito, Y TI High-speed counter-current chromatography of apple procyanidins SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE counter-current chromatography; apples; food analysis; procyanidins; tannins; polyphenols; flavonoids ID LIQUID PARTITION CHROMATOGRAPHY; COIL PLANET CENTRIFUGE; CONDENSED TANNINS; SOLID SUPPORT; IN-VIVO; FRACTIONATION; PROTEIN; PHASE; CELLS AB Apple procyanidins were separated by high-speed counter-current chromatography using a type-J multilayer coil planet centrifuge. Several two-phase solvent systems with a wide range of hydrophobicities from a non-polar hexane system to polar n-butanol systems were evaluated their performance in terms of the partition coefficient and the retention of the phase. The best separation of procyanidins B and C was achieved with a two-phase solvent system composed of n-butanol-methyl tert.-butyl ether-acetonitrile-0.1% trifluoroacetic acid (2:4:3:8) using the lower phase as a mobile at a flow-rate of 1.0 ml/min. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Analyt Chem, Hachioji, Tokyo 1920392, Japan. NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Shibusawa, Y (reprint author), Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Analyt Chem, 1432-1 Horinouchi, Hachioji, Tokyo 1920392, Japan. NR 25 TC 17 Z9 17 U1 0 U2 5 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 21 PY 2000 VL 886 IS 1-2 BP 65 EP 73 DI 10.1016/S0021-9673(00)00448-9 PG 9 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 334ME UT WOS:000088189600007 PM 10950276 ER PT J AU Tian, GL Zhang, TY Yang, FQ Ito, Y AF Tian, GL Zhang, TY Yang, FQ Ito, Y TI Separation of gallic acid from Cornus officinalis Sieb. et Zucc by high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE Cornus officinalis; counter-current chromatography; gallic acid AB Gallic acid was separated from a n-butanol extract of the fruit of Cornus officinalis Sieb. et Zucc by high-speed countercurrent chromatography in two steps using two solvent systems composed of ethyl-acetate-ethanol n-butanol-water (5:1.8:6, v/v/v) and ethyl acetate-ethanol-water (5:0.5:6, v/v/v) successively. From 1 g of n-butanol extract the method produced 60 mg of gallic acid at a purity of 97%. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Beijing Inst New Technol Applicat, Beijing 100035, Peoples R China. NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Tian, GL (reprint author), Beijing Inst New Technol Applicat, Beijing 100035, Peoples R China. NR 5 TC 37 Z9 44 U1 0 U2 9 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 21 PY 2000 VL 886 IS 1-2 BP 309 EP 312 DI 10.1016/S0021-9673(00)00480-5 PG 4 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 334ME UT WOS:000088189600030 PM 10950299 ER PT J AU Patrick, CB Krzywkowski, P Ramassamy, C Poirier, J Rapoport, SI Murphy, EJ AF Patrick, CB Krzywkowski, P Ramassamy, C Poirier, J Rapoport, SI Murphy, EJ TI Phospholipase A(2) activity is decreased selectively in the hippocampus of aged apolipoprotein E deficient mice SO NEUROSCIENCE LETTERS LA English DT Article DE long term potentiation; apolipoprotein E; phospholipase A(2); hippocampus; knockout mice; aging; Alzheimer disease; arachidonic acid ID LONG-TERM POTENTIATION; ALZHEIMERS-DISEASE; E POLYMORPHISM; TYPE-4 ALLELE; MEMORY; INDUCTION; RECEPTORS; BINDING AB Because apolipoprotein E (ApoE) deficient mice have cognitive deficits (Neurosci. Lett., 199 (1995) 1-4; Neuroscience, 92 (1999) 1273-1286; Brain Res., 752 (1997) 189-796) that may involve decreased phospholipase A(2) (PLA(2)) activity (Neuroscience, 92 (1999) 1273-1286), striatal, hippocampal, and parieto-temporal PLA(2) activity was measured in cytosol from 3 and 20-month-old ApoE deficient and control mice. Samples were homogenized and cytosol prepared by ultracentrifugation. PLA(2) activity in each cytosolic fraction was measured in triplicate using a continuous fluorometric assay (J. Neurosci. Methods, (2000) in press). In 3-month-old animals, there was a trend for decreased hippocampal PLA(2) activity between groups. In 20-month-old animals, hippocampal PLA(2) activity was significantly (P = 0.0304) decreased nearly 20% in ApoE deficient mice as compared to age-matched control mice. No differences were found in other brain regions, although activity in the striatal samples were nearly 65% less than in the other two regions. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 NIA, Sect Brain Physiol & Metab, NIH, Bethesda, MD 20892 USA. Douglas Hosp, Res Ctr, Verdun, PQ H4H 1R3, Canada. RP Murphy, EJ (reprint author), NIA, Sect Brain Physiol & Metab, NIH, Bldg 10,Room 6C103,10 Ctr Dr MSC 1582, Bethesda, MD 20892 USA. NR 20 TC 2 Z9 2 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 21 PY 2000 VL 288 IS 3 BP 211 EP 214 DI 10.1016/S0304-3940(00)01240-4 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 336RN UT WOS:000088316900012 PM 10889345 ER PT J AU Arif, JM Gairola, CG Kelloff, GJ Lubet, RA Gupta, RC AF Arif, JM Gairola, CG Kelloff, GJ Lubet, RA Gupta, RC TI Inhibition of cigarette smoke-related DNA adducts in rat tissues by indole-3-carbinol SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE indole-3-carbinol; cigarette smoke; DNA adducts; P-32-postlabeling; rat lung ID DIETARY INDOLE-3-CARBINOL; MAMMARY CARCINOGENESIS; BRASSICA VEGETABLES; LUNG TUMORIGENESIS; A/J MICE; CANCER; TUMOR; CHEMOPREVENTION; ENHANCEMENT; HEPATOCARCINOGENESIS AB Indole-3-carbinol (I3C) found in various cruciferous vegetables has been shown to exert anti-carcinogenic activity in several target organs. In this study, we have investigated the effects of I3C on cigarette smoke-related lipophilic DNA adduct formation, potentially a key step in chemical carcinogenesis. Female Sprague-Dawley rats were exposed to sidestream cigarette smoke in a whole-body exposure chamber for 6 h per day, 7 days a week for 4 weeks. Control animals received only vehicle while the intervention groups received I3C (1.36 or 3.40 mmol/kg, b.wt.) daily by gavage starting from 1 week prior to smoke initiation until the end of the experiment. Analysis of tissue DNA by nuclease Pi-mediated P-32-postlabeling showed one major and several minor smoke-related adducts in lung, trachea, heart and bladder. The high dose of I3C significantly inhibited the major adducts in lung (#5) and trachea (#3) by 55% each; minor adducts were slightly inhibited (20-40%). The low dose of I3C showed lesser degree of inhibition (30-40%) in both lung and trachea; however, it was found statistically significant in lung only. The major smoke-related adduct in bladder (#2) was strongly inhibited (> 65%) by high dose of I3C approaching adduct levels achieved in sham-exposed rats. A small but statistically significant decrease in the smoke-related DNA adduct (#5) in heart tissue was also observed by intervention with high dose I3C. Low levels (30-50 adducts/10(10) nucleotides) of I3C-derived DNA adducts were also found in all the tissues examined although their significance remains unknown. These data show significant inhibition of cigarette smoke-related DNA adducts by I3C, particularly in the lung, trachea, and bladder, (C) 2000 Published by Elsevier Science B.V. C1 Univ Kentucky, Dept Prevent Med & Environm Hlth, Lexington, KY 40536 USA. Univ Kentucky, Tobacco & Hlth Res Inst, Lexington, KY 40536 USA. NCI, Dept Chemoprevent, Invest Branch, Bethesda, MD 20892 USA. RP Gupta, RC (reprint author), Univ Kentucky, Dept Prevent Med & Environm Hlth, 354 Hlth Sci Res Bldg, Lexington, KY 40536 USA. FU NCI NIH HHS [CA-01688, CN45591] NR 47 TC 16 Z9 19 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0027-5107 J9 MUTAT RES-FUND MOL M JI Mutat. Res.-Fundam. Mol. Mech. Mutagen. PD JUL 20 PY 2000 VL 452 IS 1 BP 11 EP 18 DI 10.1016/S0027-5107(00)00010-5 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 344GA UT WOS:000088749800002 PM 10894885 ER PT J AU Loukinova, E Dong, G Enamorado-Ayalya, I Thomas, GR Chen, Z Schreiber, H Van Waes, C AF Loukinova, E Dong, G Enamorado-Ayalya, I Thomas, GR Chen, Z Schreiber, H Van Waes, C TI Growth regulated oncogene-alpha expression by murine squamous cell carcinoma promotes tumor growth, metastasis, leukocyte infiltration and angiogenesis by a host CXC receptor-2 dependent mechanism SO ONCOGENE LA English DT Article DE GRO-alpha; KC; CXCR2; chemokines; squamous cell carcinoma ID PROINFLAMMATORY CYTOKINE EXPRESSION; COLONY-STIMULATING FACTOR; FACTOR-KAPPA-B; CONSTITUTIVE ACTIVATION; ENVIRONMENT PROMOTES; CANCER-CELLS; NECK-CANCER; HUMAN HEAD; IN-VIVO; LINES AB Growth Regulated Oncogene-alpha (GRO-alpha) is an autocrine growth factor in melanoma and is a member of the C-X-C family of chemokines which promote chemotaxis of granulocytes and endothelia through binding to CXC Receptor 2, We found previously that variants of murine squamous cell carcinoma PAM 212 which grow and metastasize more rapidly in vivo constitutively express increased levels of murine GRO-alpha, designated mGRO-alpha, or KC, We have examined the possible role of mGRO-alpha. expression in malignant progression of squamous cell carcinoma PAM 212 in homologous BALB/c and BALB CXC Receptor-2 deficient mice, Transfection of the PAM 212 cell line which exhibits low expression of GRO-alpha and malignant potential with a pActin-KC vector encoding mGRO-alpha enabled isolation of PAM-KC expressing cell lines, These PAM-KC transfectants displayed an increased rate of growth and metastasis in BALB/c mice, similar to the highly malignant phenotype observed in spontaneously occurring metastatic variants. Furthermore, the PAM-KC tumors showed an increase in infiltration of host leukocytes and CD31+ blood vessels, consistent with increased CXC chemokine activity. The increased growth of PAM-KC cells was attenuated in CXCR-2 deficient mice, indicating that the increased growth was dependent in part upon host cells responsive to the CXC chemokine, Together, these results show that a CXC chemokine such as GRO-alpha can promote malignant growth of murine squamous cell carcinoma by a host CXCR-2 dependent pathway. C1 Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. Univ Chicago, Comm Immunol, Dept Pathol, Chicago, IL 60637 USA. RP Van Waes, C (reprint author), Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bldg 10,Rm 5D55,MSC-1419, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA-22677]; NIDCD NIH HHS [Z01-DC-00016] NR 25 TC 122 Z9 130 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUL 20 PY 2000 VL 19 IS 31 BP 3477 EP 3486 DI 10.1038/sj.onc.1203687 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 337FJ UT WOS:000088346500004 PM 10918606 ER PT J AU Kamei, A Tamaki, S Taniyama, H Takamura, S Nishimura, Y Kagawa, Y Uno-Furuta, S Kaito, M Kim, G Toda, M Matsuura, Y Miyamura, T Adachi, Y Yasutomi, Y AF Kamei, A Tamaki, S Taniyama, H Takamura, S Nishimura, Y Kagawa, Y Uno-Furuta, S Kaito, M Kim, G Toda, M Matsuura, Y Miyamura, T Adachi, Y Yasutomi, Y TI Induction of hepatitis C virus-specific cytotoxic T lymphocytes in mice by an intrahepatic inoculation with an expression plasmid SO VIROLOGY LA English DT Article DE CTL; DNA vaccine; animal model; HCV ID TRANSGENIC MICE; NON-A; STRUCTURAL PROTEINS; CTL RESPONSES; LIVER-CELLS; CDNA; MOUSE; CHIMPANZEES; INFECTION; ANTIGENS AB We assessed the possibility of intrahepatic inoculation with a plasmid encoding hepatitis C virus (HCV) proteins to elicit HCV-specific cytotoxic T lymphocyces (CTL) in mice as a conventional animal model of HCV infection. BALB/c mice were intrahepatically or intramuscularly inoculated with an expression plasmid DNA encoding HCV structural proteins under the control of the elongation factor 1-alpha promoter. Expressions of HCV-core protein and envelope proteins (E1 and E2) in hepatocytes were detected immunohistochemically 6 days after inoculation. CTL responses were examined using target cells either pulsed with a specific peptide or infected with a recombinant vaccinia virus expressing HCV structural protein. Both intrahepatically and intramuscularly DNA-inoculated mice developed CD8(+), MHC class I-restricted CTL responses that recognized the peptide pulsed as well as HCV proteins expressing target cells. These studies demonstrated the usefulness of a murine model of HCV infection induced by direct intrahepatic DNA inoculation for understanding the immunopathogenic mechanisms in HCV infection. (C) 2000 Academic Press. C1 Mie Univ, Sch Med, Dept Bioregulat, Tsu, Mie 5148507, Japan. Mie Univ, Sch Med, Dept Internal Med 3, Tsu, Mie 5148507, Japan. Rakuno Gakuen Univ, Dept Vet Pathol, Ebetsu, Hokkaido 0690836, Japan. NIAID, Dept Virol 2, Tokyo 1628640, Japan. RP Yasutomi, Y (reprint author), Mie Univ, Sch Med, Dept Bioregulat, Tsu, Mie 5148507, Japan. NR 27 TC 3 Z9 4 U1 0 U2 0 PU ACADEMIC PRESS INC 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 2000 VL 273 IS 1 BP 120 EP 126 DI 10.1006/viro.2000.0403 PG 7 WC Virology SC Virology GA 340JF UT WOS:000088528800014 PM 10891414 ER PT J AU Yancey, KB Egan, CA AF Yancey, KB Egan, CA TI Pemphigoid: Clinical, histologic, immunopathologic, and therapeutic considerations SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID GESTATIONIS AUTOANTIBODIES RECOGNIZE; EPIDERMOLYSIS-BULLOSA ACQUISITA; HERPES-GESTATIONIS; EXTRACELLULAR DOMAIN; IGG AUTOANTIBODIES; SUBCLASS DISTRIBUTION; BASEMENT-MEMBRANE; BP180 ECTODOMAIN; PASSIVE TRANSFER; HG-FACTOR AB Autoimmune blistering diseases are generally distinct entities characterized by relatively consistent clinical, histologic, and immunopathologic findings. These disorders may cause impaired adhesion of epidermis to epidermal basement membrane (eg, the pemphigoid group of disorders [bullous, gestational, and mucous membrane]) or impaired adhesion of epidermal cells to each other (eg, the pemphigus group of disorders). Recent studies have shown that these disorders are characterized by autoantibodies that often display pathogenic (ie, blister-forming) activity in passive transfer models. Interestingly, the autoantigens targeted by these patients' autoantibodies represent important structural proteins that promote cell matrix (eg, pemphigoid) or cell-to-cell (eg, pemphigus) adhesion in skin. Autoimmune blistering diseases are characterized by substantial morbidity (pruritus, pain, disfigurement), and in some instances, mortality (secondary to loss of epidermal barrier function). Treatment with systemic immunosuppressives has reduced morbidity and mortality in patients with these diseases. C1 NCI, Dermatol Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Yancey, KB (reprint author), NCI, Dermatol Branch, Div Clin Sci, NIH, Bldg 10,Room 12N238,10 Ctr Dr,MSC 1908, Bethesda, MD 20892 USA. NR 62 TC 58 Z9 61 U1 0 U2 1 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 19 PY 2000 VL 284 IS 3 BP 350 EP 356 DI 10.1001/jama.284.3.350 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 333GR UT WOS:000088123000028 PM 10891967 ER PT J AU Clark, CM Fradkin, JE Hiss, RG Lorenz, RA Vinicor, F Warren-Boulton, E AF Clark, CM Fradkin, JE Hiss, RG Lorenz, RA Vinicor, F Warren-Boulton, E TI Promoting early diagnosis and treatment of type 2 diabetes - The National Diabetes Education Program SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Editorial Material ID NON-HISPANIC WHITES; GLYCEMIC CONTROL; CARE; MANAGEMENT; NIDDM; MELLITUS; ADULTS; POPULATION; BENEFITS; QUALITY C1 NIDDKD, NIH, Bethesda, MD 20892 USA. Regenstrief Inst Hlth Care, Indianapolis, IN 46202 USA. Univ Michigan Hlth Syst, Michigan Diabet Res & Training Ctr, Demonstrat & Educ Div, Ann Arbor, MI USA. Univ Illinois, Coll Med, Dept Pediat, Peoria, IL 61656 USA. Ctr Dis Control & Prevent, Div Diabet Translat, Atlanta, GA USA. Hager Sharp, Washington, DC USA. RP Clark, CM (reprint author), Natl Diabet Educ Program, Box JAMA Reprint,1 Diabet Way, Bethesda, MD 20892 USA. NR 32 TC 42 Z9 44 U1 0 U2 2 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 19 PY 2000 VL 284 IS 3 BP 363 EP 365 DI 10.1001/jama.284.3.363 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 333GR UT WOS:000088123000030 PM 10891969 ER PT J AU Wacholder, S Rothman, N Caporaso, N AF Wacholder, S Rothman, N Caporaso, N TI Population stratification in epidemiologic studies of common genetic variants and cancer: Quantification of bias SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID ACETYLATOR PHENOTYPE; RECEPTOR GENE; ASSOCIATION; ALCOHOLISM; ADMIXTURE; RISK; AGE AB Background: Some critics argue that bias from population stratification (the mixture of individuals from heterogeneous genetic backgrounds) undermines the credibility of epidemiologic studies designed to estimate the association between a genotype and the risk of disease. We investigated the degree of bias likely from population stratification in U.S. studies of cancer among non-Hispanic Caucasians of European origin. Methods: An expression of the confounding risk ratio-the ratio of the effect of the genetic factor on risk of disease with and without adjustment for ethnicity-is used to measure the potential relative bias from population stratification. We first use empirical data on the frequency of the N-acetyltransferase (NAT2) slow acetylation genotype and incidence rates of male bladder cancer and female breast cancer in non-Hispanic U.S, Caucasians with ancestries from eight European countries to assess the bias in a hypothetical population-based U.S. study that does not take ethnicity into consideration. Then, we provide theoretical calculations of the bias over a large range of allele frequencies and disease rates. Results: Ignoring ethnicity leads to a bias of 1% or less in our empirical studies of NAT2. Furthermore, evaluation of a wide range of allele frequencies and representative ranges of cancer rates that exist across European populations shows that the risk ratio is biased by less than 10% in U.S. studies except under extreme conditions. We note that the bias decreases as the number of ethnic strata increases. Conclusions: There will be only a small bias from population stratification in a well-designed case-control study of genetic factors that ignores ethnicity among non-Hispanic U.S. Caucasians of European origin. Further work is needed to estimate the effect of population stratification within other populations. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Wacholder, S (reprint author), NIH, EPS 8046,6120 Execut Blvd, Bethesda, MD 20892 USA. NR 28 TC 316 Z9 319 U1 0 U2 3 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD JUL 19 PY 2000 VL 92 IS 14 BP 1151 EP 1158 DI 10.1093/jnci/92.14.1151 PG 8 WC Oncology SC Oncology GA 334UJ UT WOS:000088203800010 PM 10904088 ER PT J AU Travis, LB Andersson, M Gospodarowicz, M van Leeuwen, FE Bergfeldt, K Lynch, CF Curtis, RE Kohler, BA Wiklund, T Storm, H Holowaty, E Hall, P Pukkala, E Sleijfer, DT Clarke, EA Boice, JD Stovall, M Gilbert, E AF Travis, LB Andersson, M Gospodarowicz, M van Leeuwen, FE Bergfeldt, K Lynch, CF Curtis, RE Kohler, BA Wiklund, T Storm, H Holowaty, E Hall, P Pukkala, E Sleijfer, DT Clarke, EA Boice, JD Stovall, M Gilbert, E TI Treatment-associated leukemia following testicular cancer SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID NON-HODGKINS-LYMPHOMA; GERM-CELL CANCER; SECONDARY LEUKEMIA; OVARIAN-CANCER; RISK; CHEMOTHERAPY; ETOPOSIDE; CISPLATIN; SEMINOMA; THERAPY AB Background: Men with testicular cancer are at an increased risk of leukemia, but the relationship to prior treatments is not well characterized. The purpose of our study was to describe the risk of leukemia following radiotherapy and chemotherapy for testicular cancer. Methods: Within a population-based cohort of 18 567 patients diagnosed with testicular cancer (from 1970 through 1993), a case-control study of leukemia was undertaken, Radiation dose to active bone marrow and type and cumulative amount of cytotoxic drugs were compared between 36 men who developed leukemia and 106 matched control patients without leukemia. Conditional logistic regression was used to estimate the relative risk of leukemia associated with specific treatments. All P values are two-sided. Results: Radiotherapy (mean dose to active bone marrow, 12.6 Gy) without chemotherapy was associated with a threefold elevated risk of leukemia. Risk increased with increasing dose of radiation to active bone marrow (P for trend = .02), with patients receiving radiotherapy to the chest as well as to the abdominal/pelvic fields accounting for much of the risk at higher doses. Radiation dose to active bone marrow and the cumulative dose of cisplatin (P for trend = .001) were both predictive of excess leukemia risk in a model adjusted for all treatment variables. The estimated relative risk of leukemia at a cumulative dose of 650 mg cisplatin, which is commonly administered in current testicular cancer treatment regimens, was 3.2 (95% confidence interval = 1.5-8.4); larger doses (1000 mg) were linked with statistically significant sixfold increased risks. Conclusions: Past treatments for testicular cancer are associated with an increased risk of leukemia, with evidence for dose-response relationships for both radiotherapy and cisplatin-based chemotherapy. Statistically nonsignificant excesses are estimated for current radiotherapy regimens limited to the abdomen and pelvis: Among 10 000 patients given a treatment dose of 25 Gy and followed for 15 years, an excess of nine leukemias is predicted; cisplatin-based chemotherapy (dose, 650 mg) might result in 16 cases of leukemia. The survival advantage provided by current radiotherapy and chemotherapy regimens for testicular cancer far exceeds the small absolute risk of leukemia. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Danish Canc Soc, Copenhagen, Denmark. Univ Toronto, Princess Margaret Hosp, Toronto, ON, Canada. Netherlands Canc Inst, Amsterdam, Netherlands. Karolinska Inst, Stockholm, Sweden. Univ Iowa, Iowa City, IA USA. Dept Hlth, Trenton, NJ USA. Senior Serv, Trenton, NJ USA. Univ Helsinki, Cent Hosp, FIN-00014 Helsinki, Finland. Canc Care Ontario, Toronto, ON, Canada. Finnish Canc Registry, FIN-00170 Helsinki, Finland. Univ Groningen, NL-9700 AB Groningen, Netherlands. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. RP Travis, LB (reprint author), NIH, Execut Plaza S,Suite 7086, Bethesda, MD 20892 USA. RI Bergfeldt, Kjell/C-3581-2012 NR 39 TC 149 Z9 152 U1 0 U2 2 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD JUL 19 PY 2000 VL 92 IS 14 BP 1165 EP 1171 DI 10.1093/jnci/92.14.1165 PG 7 WC Oncology SC Oncology GA 334UJ UT WOS:000088203800012 PM 10904090 ER PT J AU Whiteman, DC Murphy, MFG Cook, LS Cramer, DW Hartge, P Marchbanks, PA Nasca, PC Ness, RB Purdie, DM Risch, HA AF Whiteman, DC Murphy, MFG Cook, LS Cramer, DW Hartge, P Marchbanks, PA Nasca, PC Ness, RB Purdie, DM Risch, HA TI Multiple births and risk of epithelial ovarian cancer SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID DIZYGOTIC TWINS; INCESSANT OVULATION; HORMONE LEVELS; UNITED-STATES; MOTHERS; PROGESTERONE; PREGNANCIES; PARITY; WOMEN AB Background and Methods: Prevailing hypotheses about the causes of ovarian carcinogenesis predict that women with a history of multiple births (twins, triplets, etc.) should be at increased risk of epithelial ovarian cancer. However, the scant available evidence suggests that they may actually be at lower risk. To resolve this issue, we pooled data from eight studies involving 2859 parous women with epithelial ovarian cancer (case patients) and 7434 parous women without ovarian cancer (control women). In addition to assessing their history of multiple births (and the sex of the children, where available), we obtained information on age, parity, oral contraceptive use, and other reproductive factors for each woman. Details of tl:mor histology were available for all case patients. We estimated the relative risks of various histologic types of ovarian cancers associated with multiple births by using multivariable logistic regression analysis, adjusting for matching and confounding variables. Results: Among these parous women, 73 case patients (2.6%) and 257 control women (3.5%) had a history of multiple births. The adjusted summary odds ratio (OR) for developing all types of epithelial ovarian cancer that are associated with multiple births was 0.81 (95% confidence interval [CI] = 0.61-1.08), We found no evidence that risks associated with multiple births differed among women with borderline or invasive tumors and among women with same-sex and opposite-sex offspring from multiple births. The risk reductions appeared specific for nonmucinous tumors (n = 2453; summary adjusted OR = 0.71 [95% CI = 0.52-0.98]); in contrast, associations with mucinous tumors (n = 406) were heterogeneous across studies. Conclusions: Parous women with nonmucinous ovarian cancer are no more likely to have a history of multiple births than other parous women, counter to the predictions of current hypotheses for causes of ovarian cancer. C1 Univ Oxford, Imperial Canc Res Fund, Gen Practice Res Grp, Oxford OX1 2JD, England. Univ Calgary, Calgary, AB T2N 1N4, Canada. Harvard Univ, Sch Med, Boston, MA USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Fertil Epidemiol Sect, Atlanta, GA USA. Univ Massachusetts, Amherst, MA 01003 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. Queensland Inst Med Res, Brisbane, Qld 4006, Australia. Yale Univ, Sch Med, New Haven, CT USA. RP Whiteman, DC (reprint author), Queensland Inst Med Res, Epidemiol & Populat Hlth Unit, PO Royal Brisbane Hosp, Brisbane, Qld 4029, Australia. RI Whiteman, David/P-2728-2014 OI Whiteman, David/0000-0003-2563-9559 NR 39 TC 36 Z9 36 U1 0 U2 1 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD JUL 19 PY 2000 VL 92 IS 14 BP 1172 EP 1177 DI 10.1093/jnci/92.14.1172 PG 6 WC Oncology SC Oncology GA 334UJ UT WOS:000088203800013 PM 10904091 ER PT J AU Senderowicz, AM Sausville, EA AF Senderowicz, AM Sausville, EA TI Re: Preclinical and clinical development of cyclin-dependent kinase modulators - Response SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Letter ID CELLS; FLAVOPIRIDOL; MURINE C1 Natl Inst Dental & Craniofacial Res, Oral & Pharyngeal Canc Branch, Mol Therapeut Unit, Bethesda, MD USA. NCI, Div Canc Treatment & Diag, Dev Therapeut Program, Clin Trial Unit, Bethesda, MD 20892 USA. RP Senderowicz, AM (reprint author), NIH, Bldg 30,Rm 211, Bethesda, MD 20892 USA. NR 6 TC 4 Z9 4 U1 0 U2 1 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD JUL 19 PY 2000 VL 92 IS 14 BP 1185 EP 1185 DI 10.1093/jnci/92.14.1185 PG 1 WC Oncology SC Oncology GA 334UJ UT WOS:000088203800019 ER PT J AU Selengut, JD Levine, RL AF Selengut, JD Levine, RL TI MDP-1: A novel eukaryotic magnesium-dependent phosphatase SO BIOCHEMISTRY LA English DT Article ID PROTEIN-TYROSINE PHOSPHATASES; P-TYPE ATPASE; CRYSTAL-STRUCTURE; ACID-PHOSPHATASE; IDENTIFICATION; SUPERFAMILY; MECHANISM; CATALYSIS; VANADATE; SITE AB We report here the purification, cloning, expression, and characterization of a novel phosphatase, MDP-1. In the course of investigating the reported acid phosphatase activity of carbonic anhydrase III preparations, several discrete phosphatases were discerned. One of these, a magnesium-dependent species of 18.6 kDa, was purified to homogeneity and yielded several peptide sequences from which the parent gene was identified by database searching. Although orthologous genes were identified in fungi and plants as well as mammalian species, there was no apparent homology to any known family of phosphatases. The enzyme was expressed in Escherichia coli with a fusion tag and purified by affinity methods. The recombinant enzyme showed magnesium-dependent acid phosphatase activity comparable to the originally isolated rabbit protein. The enzyme catalyzes the rapid hydrolysis of p-nitrophenyl phosphate, ribose-5-phosphate, and phosphotyrosine. The selectivity for phosphotyrosine over phosphoserine or phosphothreonine is considerable, but the enzyme did not show activity toward five phosphotyrosine-containing peptides. None of the various substrates assayed (including various nucleotide, sugar, amino acid and peptide phosphates, phosphoinositides, and phosphodiesters) exhibited KM values lower than 1 mM, and many showed negligible rates of hydrolysis. The enzyme is inhibited by vanadate and fluoride but not by azide, cyanide, calcium, lithium, or tartaric acid. Chemical labeling, refolding, dialysis, and mutagenesis experiments suggest that the enzymatic mechanism is not dependent on cysteine, histidine, or nonmagnesium metal ions. In recognition of these observations, the enzyme has been given the name magnesium-dependent phosphatase-1 (MDP-1). C1 NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Selengut, JD (reprint author), NHLBI, Biochem Lab, NIH, Bldg 3-118,3 Ctr Dr, Bethesda, MD 20892 USA. EM selengut@nih.gov RI Levine, Rodney/D-9885-2011 NR 24 TC 29 Z9 30 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 18 PY 2000 VL 39 IS 28 BP 8315 EP 8324 DI 10.1021/bi0005052 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 337TB UT WOS:000088376800026 PM 10889041 ER PT J AU Stobie, L Gurunathan, S Prussin, C Sacks, DL Glaichenhaus, N Wu, CY Seder, RA AF Stobie, L Gurunathan, S Prussin, C Sacks, DL Glaichenhaus, N Wu, CY Seder, RA TI The role of antigen and IL-12 in sustaining Th1 memory cells in vivo: IL-12 is required to maintain memory/effector Th1 cells sufficient to mediate protection to an infectious parasite challenge SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID INTERFERON-GAMMA PRODUCTION; CD4(+) T-CELLS; LEISHMANIA-MAJOR; IFN-GAMMA; CUTANEOUS LEISHMANIASIS; IMMUNE-RESPONSE; POLARIZED TH1; MICE; INTERLEUKIN-12; EXPRESSION AB IL-12 plays a central role in both the induction and magnitude of a primary Th1 response. A critical question in designing vaccines for diseases requiring Th1 immunity such as Mycobacterium tuberculosis and Leishmania major is the requirements to sustain memory/effector Th1 cells in vivo. This report examines the role of IL-12 and antigen in sustaining Th1 responses sufficient for protective immunity to L. major after vaccination with LACK protein (LP) plus rIL-12 and LACK DNA. It shows that, after initial vaccination with LP plus rIL-12, supplemental boosting with either LP or rIL-12 is necessary but not sufficient to fully sustain long-term Th1 immunity. Moreover, endogenous IL-12 is also shown to be required for the induction, maintenance, and effector phase of the Th1 response after LACK DNA vaccination. Finally, IL-12 is required to sustain Th1 cells and control parasite growth in susceptible and resistant strains of mice during primary and secondary infection. Taken together, these data show that IL-12 is essential to sustain a sufficient number of memory/effector Th1 cells generated in vivo to mediate long-term protection to an intracellular pathogen. C1 NIAID, Clin Immunol Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. NIAID, Howard Hughes Med Inst, NIH, Bethesda, MD 20892 USA. NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. CNRS, Inst Pharmacol Mol & Cellulaire, UPR411, F-06560 Valbonne, France. RP Seder, RA (reprint author), NIAID, Clin Immunol Sect, Clin Invest Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Prussin, Calman/0000-0002-3917-3326 NR 34 TC 137 Z9 146 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 18 PY 2000 VL 97 IS 15 BP 8427 EP 8432 DI 10.1073/pnas.160197797 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335YF UT WOS:000088273900045 PM 10890924 ER PT J AU Villeneuve, S Souchon, H Riottot, MM Mazie, JC Lei, P Glaudemans, CPJ Kovac, P Fournier, JM Alzari, PM AF Villeneuve, S Souchon, H Riottot, MM Mazie, JC Lei, P Glaudemans, CPJ Kovac, P Fournier, JM Alzari, PM TI Crystal structure of an anti-carbohydrate antibody directed against Vibrio cholerae 01 in complex with antigen: Molecular basis for serotype specificity SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID MONOCLONAL-ANTIBODY; ANGSTROM RESOLUTION; COMBINING SITES; LIGHT CHAIN; OGAWA; O1; IMMUNOGLOBULIN; RECOGNITION; INABA; LIPOPOLYSACCHARIDE AB The crystal structure of the murine Fab S-20-4 from a protective anti-cholera Ab specific for the lipopolysaccharide Ag of the Ogawa serotype has been determined in its unliganded form and in complex with synthetic fragments of the Ogawa O-specific polysaccharide (O-SP). The upstream terminal O-SP monosaccharide is shown to be the primary antigenic determinant. Additional perosamine residues protrude outwards from the Ab surface and contribute only marginally to the binding affinity and specificity. A complementary water-excluding hydrophobic interface and five Ab-AS hydrogen bonds are crucial for carbohydrate recognition. The structure reported here explains the serotype specificity of anti-Ogawa Abs and provides a rational basis toward the development of a synthetic carbohydrate-based anti-cholera vaccine. C1 Inst Pasteur, Unite Biochim & Struct, CNRS URA 2185, F-75724 Paris 15, France. Inst Pasteur, Unite Cholera & Vibrat, F-75724 Paris, France. Inst Pasteur, Unite Immunol Struct, F-75724 Paris, France. Inst Pasteur, Lab Ingn Anticorps, F-75724 Paris, France. NIDDK, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Alzari, PM (reprint author), Inst Pasteur, Unite Biochim & Struct, CNRS URA 2185, 25 Rue Dr Roux, F-75724 Paris 15, France. EM alzari@pasteur.fr NR 42 TC 78 Z9 81 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 18 PY 2000 VL 97 IS 15 BP 8433 EP 8438 DI 10.1073/pnas.060022997 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335YF UT WOS:000088273900046 PM 10880560 ER PT J AU Rathore, D McCutchan, TF AF Rathore, D McCutchan, TF TI Role of cysteines in Plasmodium falciparum circumsporozoite protein: Interactions with heparin can rejuvenate inactive protein mutants SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ANTI-SPOROZOITE ANTIBODIES; HUMAN MALARIA PARASITE; REGION-II-PLUS; THROMBOSPONDIN; BINDING; CELL; PROPERDIN; SEQUENCE; MOTIFS; PROTEOGLYCANS AB Various pathogenic bacteria, viruses, and protozoan bind to glycosaminoglycan-based receptors on host cells and initiate an infection. Sporozoites of Plasmodium predominantly express circumsporozoite (CS) protein on their surface, which binds to heparan sulfate proteoglycans on liver cell surface that subsequently leads to malaria. Here we show that the interaction of free heparin with this parasite ligand has the potential to be a critical component of invasion. CS protein of P. falciparum contains four cysteines at positions 361. 365, 396. and 401. In this study, all four cysteine residues were mutagenized to alanine both individually and in different combinations. Conversion of cysteine 396 to alanine (protein CS3) led to a 10-fold increase in the binding activity of the protein to HepG2 cells. Replacement of cysteines at positions 361, 365. and 401 either alone or in different combinations led to a near total loss of binding. Surprisingly, activity in these inactive mutants could be effectively restored in the presence of submolar concentrations of heparin. Heparin also up-regulated binding of CS3 at submolar concentrations with respect to the protein but downregulated binding when present in excess. Given the significantly different concentrations of heparin in different organs of the host and the in vitro results described here one can consider in vivo ramifications of this phenomenon for pathogen targeting of specific organs and for the functional effects of antigenic variation on receptor ligand interaction. C1 NIAID, Growth & Dev Sect, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP McCutchan, TF (reprint author), NIAID, Growth & Dev Sect, Parasit Dis Lab, NIH, Room 126,Bldg 4,4 Ctr Dr,MSC 0425,9000 Rockville, Bethesda, MD 20892 USA. NR 27 TC 23 Z9 23 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 18 PY 2000 VL 97 IS 15 BP 8530 EP 8535 DI 10.1073/pnas.140224597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335YF UT WOS:000088273900063 PM 10890903 ER PT J AU Tsutsumi, Y Onda, M Nagata, S Lee, B Kreitman, RJ Pastan, I AF Tsutsumi, Y Onda, M Nagata, S Lee, B Kreitman, RJ Pastan, I TI Site-specific chemical modification with polyethylene glycol of recombinant immunotoxin anti-Tac(Fv)-PE38 (LMB-2) improves antitumor activity and reduces animal toxicity and immunogenicity SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE Pseudomonas exotoxin; cancer therapy; immunotherapy; pharmacokinetics; liver toxicity ID SURFACE-EXPOSED RESIDUES; NECROSIS-FACTOR-ALPHA; EXOTOXIN-A MUTANTS; PSEUDOMONAS EXOTOXIN; COMPLETE REGRESSION; CYSTEINE RESIDUES; MOLECULAR DESIGN; HUMAN CARCINOMA; FV FRAGMENT; MICE AB Chemical modification of proteins with polyethylene glycol (PEGy-lation) can increase plasma half-lives, stability, and therapeutic potency. To make a PEGylated recombinant immunotoxin with improved therapeutic properties, we prepared a mutant of anti-Tac(Fv)-PE38 (LMB-2), a recombinant immunotoxin composed of a single-chain Fv fragment of the anti-human Tac monoclonal antibody to the IL-2 receptor cu subunit fused to a 38-kDa fragment of Pseudomonas exotoxin. For site-specific PEGylation of LMB-2, one cysteine residue was introduced into the peptide connector (ASGCGPE) between the Fv and the toxin. This mutant LMB-2 (cys1-LMB-2), which retained full cytotoxic activity, was then site-specifically conjugated with 5 or 20 kDa of polyethylene glycol-maleimide. When compared with unmodified LMB-2, both PEGylated immunotoxins showed similar cytotoxic activities in vitro but superior stability at 37 degrees C in mouse serum, a 5- to 8-fold increase in plasma half-lives in mice, and a 3- to 4-fold increase in antitumor activity. This was accompanied by a substantial decrease in animal toxicity and immunogenicity. Site-specific PEGylation of recombinant immunotoxins may increase their therapeutic potency in humans. C1 NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP NCI, Mol Biol Lab, Div Basic Sci, NIH, Bldg 37,Room 4E16,37 Convent Dr MSC 4255, Bethesda, MD 20892 USA. EM pasta@helix.nih.gov NR 26 TC 165 Z9 172 U1 0 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 18 PY 2000 VL 97 IS 15 BP 8548 EP 8553 DI 10.1073/pnas.140210597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335YF UT WOS:000088273900066 PM 10890891 ER PT J AU Wong, ML Xie, BX Beatini, N Phu, P Marathe, S Johns, A Golds, PW Hirsch, E Williams, KJ Licinio, J Tabas, I AF Wong, ML Xie, BX Beatini, N Phu, P Marathe, S Johns, A Golds, PW Hirsch, E Williams, KJ Licinio, J Tabas, I TI Acute systemic inflammation up-regulates secretory sphingomyelinase in vivo: A possible link between inflammatory cytokines and atherogenesis SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID LOW-DENSITY-LIPOPROTEIN; INTERLEUKIN-1 RECEPTOR ANTAGONIST; TO-RETENTION HYPOTHESIS; CHOLESTEROL EFFLUX; ATHEROSCLEROTIC LESIONS; MYOCARDIAL-INFARCTION; EXTRACELLULAR-MATRIX; CULTURED FIBROBLASTS; SIGNAL TRANSDUCTION; ENDOTHELIAL-CELLS AB Inflammation plays a critical role in atherogenesis, yet the mediators linking inflammation to specific atherogenic processes remain to be elucidated. One such mediator may be secretory sphingomyelinase (S-SMase), a product of the acid sphingomyelinase gene. The secretion of S-SMase by cultured endothelial cells is induced by inflammatory cytokines, and in vivo data have implicated S-SMase in subendothelial lipoprotein aggregation, macrophage foam cell formation, and possibly other atherogenic processes. Thus, the goal of this study was to seek evidence for S-SMase regulation in vivo during a physiologically relevant inflammatory response. First, wild-type mice were injected with saline or lipopolysaccharide (LPS) as a model of acute systemic inflammation. Serum S-SMase activity 3 h postinjection was increased 2- to 2.5-fold by LPS (P < 0.01). To determine the role of IL-1 in the LPS response, we used IL-1 converting enzyme knockout mice, which exhibit deficient IL-1 bioactivity. The level of serum S-SMase activity in LPS-injected IL-1 converting enzyme knockout mice was approximate to 35% less than that in identically treated wild-type mice (P < 0.01). In LPS-injected IL-1-receptor antagonist knockout mice, which have an enhanced response to IL-1, serum S-SMase activity was increased 1.8-fold compared with LPS-injected wild-type mice (P < 0.01). Finally, when wild-type mice were injected directly with IL-1 beta, tumor necrosis factor or, or both, serum S-SMase activity increased 1.6-, 2.3-, and 2.9-fold, respectively (P < 0.01). These data show regulation of S-SMase activity in vivo and they wise the possibility that local stimulation of S-SMase may contribute to the effects of inflammatory cytokines in atherosclerosis. C1 Columbia Univ, Dept Med, New York, NY 10032 USA. Univ Calif Los Angeles, Sch Med, Dept Psychiat & Biobehav Sci, Los Angeles, CA 90095 USA. Columbia Univ, Dept Anat, New York, NY 10032 USA. Columbia Univ, Dept Cell Biol, New York, NY 10032 USA. Columbia Univ, Dept Obstet & Gynecol, New York, NY 10032 USA. NIMH, Clin Neuroendocrinol Branch, Intramural Res Program, NIH, Bethesda, MD 20892 USA. Berlex Biosci, Richmond, CA 94804 USA. Thomas Jefferson Univ, Dorrance H Hamilton Res Labs, Div Endocrinol Diabet & Metab Dis, Philadelphia, PA 19107 USA. RP Tabas, I (reprint author), Columbia Univ, Dept Med, 630 W 168th St, New York, NY 10032 USA. RI Wong, Ma-Li/D-7903-2011; Licinio, Julio/L-4244-2013 OI Licinio, Julio/0000-0001-6905-5884 FU NHLBI NIH HHS [HL56984, P50 HL056984] NR 71 TC 98 Z9 101 U1 2 U2 4 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 18 PY 2000 VL 97 IS 15 BP 8681 EP 8686 DI 10.1073/pnas.150098097 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335YF UT WOS:000088273900089 PM 10890909 ER PT J AU Ma, JY Weiss, E Kyle, DE Ziffer, H AF Ma, JY Weiss, E Kyle, DE Ziffer, H TI Acid catalyzed Michael additions to artemisitene SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID SUBSEQUENT CONVERSION; CONJUGATE ADDITION; ANALOGS; HOMOLOGATION; DERIVATIVES; QINGHAOSU AB A series of 14-substituted-artemisinin and 9-epiartemisinin derivatives was prepared by a titanium-tetrachloride catalyzed addition of trimethylsilyl enol ethers to artemisitene. Several compounds were four to seven times more active than artemisinin against Plasmodium falciparum. (C) 2000 Published by Elsevier Science Ltd. All rights reserved. C1 NIDDK, Chem Phys Lab, Bethesda, MD 20892 USA. NIDDK, Bioorgan Chem Lab, Bethesda, MD 20892 USA. Walter Reed Army Med Ctr, Walter Reed Army Inst Res, Dept Parasitol, Washington, DC 20307 USA. RP Ziffer, H (reprint author), NIDDK, Chem Phys Lab, Bethesda, MD 20892 USA. NR 13 TC 25 Z9 26 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD JUL 17 PY 2000 VL 10 IS 14 BP 1601 EP 1603 DI 10.1016/S0960-894X(00)00297-3 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 333UG UT WOS:000088148700027 PM 10915061 ER PT J AU Lobachev, KS Stenger, JE Kozyreva, OG Jurka, J Gordenin, DA Resnick, MA AF Lobachev, KS Stenger, JE Kozyreva, OG Jurka, J Gordenin, DA Resnick, MA TI Inverted Alu repeats unstable in yeast are excluded from the human genome SO EMBO JOURNAL LA English DT Article DE Alu repeats; human genome stability; inverted repeats; recombination; yeast ID NUCLEOTIDE EXCISION-REPAIR; DNA-POLYMERASE-DELTA; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; PALINDROMIC SEQUENCE; MUTATION AVOIDANCE; TRANSPOSON TN5; RECOMBINATION; GENE; INSTABILITY AB The nearly one million Alu repeats in human chromosomes are a potential threat to genome integrity. Alus form dense clusters where they frequently appear as inverted repeats, a sequence motif known to cause DNA rearrangements in model organisms. Using a yeast recombination system, we found that inverted Alu pairs can be strong initiators of genetic instability. The highly recombinagenic potential of inverted Alu pairs was dependent on the distance between the repeats and the level of sequence divergence. Even inverted Alus that were 86% homologous could efficiently stimulate recombination when separated by <20 bp, This stimulation was independent of mismatch repair. Mutations in the DNA metabolic genes RAD27 (FEN1), POL3 (polymerase delta) and MMS19 destabilized widely separated and diverged inverted Alus, Having defined factors affecting inverted Alu repeat stability in yeast, we analyzed the distribution of Alu pairs in the human genome. Closely spaced, highly homologous inverted Alus are rare, suggesting that they are unstable in humans. Alu pairs were identified that are potential sites of genetic change. C1 NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. Genet Informat Res Inst, Sunnyvale, CA 94089 USA. RP Resnick, MA (reprint author), NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. OI Gordenin, Dmitry/0000-0002-8399-1836 NR 52 TC 90 Z9 91 U1 1 U2 1 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 17 PY 2000 VL 19 IS 14 BP 3822 EP 3830 DI 10.1093/emboj/19.14.3822 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 338WV UT WOS:000088447000030 PM 10899135 ER PT J AU Du, QZ Wu, PD Ito, Y AF Du, QZ Wu, PD Ito, Y TI Low speed rotary countercurrent chromatography using a convoluted multilayer helical tube for industrial separation SO ANALYTICAL CHEMISTRY LA English DT Article AB An industrial-scale countercurrent chromatographic system was developed using a slowly rotating helical device which can be automated and left unattended during the operation. It uses a multilayer coiled column consisting of long convoluted Teflon tubing of 0.85 cm i.d., which provides sufficient retention of the stationary phase under a relatively high flow rate, The column is mounted on a seal-less continuous-flow rotary device which eliminates various complications such as leakage and clogging as often caused by the use of a conventional rotary seal. Using a multilayer coiled column with a 10-L capacity, 150 g of crude tea extract was successfully purified, yielding 40 g of epigallocatechin Gallate of over 92.7% purity at a recovery rate of 82.6%. The present system can be scaled up for a kilogram-scale separation. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. Chinese Acad Agr Sci, Tea Res Inst, Hangzhou 310008, Zhejiang, Peoples R China. Zhejiang Univ, Dept Chem Engn, Hangzhou 310027, Zhejiang, Peoples R China. RP Ito, Y (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 7 TC 44 Z9 46 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0003-2700 J9 ANAL CHEM JI Anal. Chem. PD JUL 15 PY 2000 VL 72 IS 14 BP 3363 EP 3365 DI 10.1021/ac991423q PG 3 WC Chemistry, Analytical SC Chemistry GA 337FQ UT WOS:000088347100053 ER PT J AU Suprynowicz, FA Groigno, L Whitaker, M Miller, FJ Sluder, G Sturrock, J Whalley, T AF Suprynowicz, FA Groigno, L Whitaker, M Miller, FJ Sluder, G Sturrock, J Whalley, T TI Activation of protein kinase C alters p34(cdc2) phosphorylation state and kinase activity in early sea urchin embryos by abolishing intracellular Ca2+ transients SO BIOCHEMICAL JOURNAL LA English DT Article DE cyclin B; embryonic cell cycle; Ins(1,4,5)P-3; mitosis; phorbol esters ID NUCLEAR-ENVELOPE BREAKDOWN; GERMINAL VESICLE BREAKDOWN; GROWTH-FACTOR RECEPTOR; CELL-DIVISION CYCLE; PHOSPHOLIPASE-C; PHORBOL ESTER; FREE CALCIUM; INOSITOL TRISPHOSPHATE; MOUSE OOCYTES; XENOPUS EGGS AB The p34(cdc2) protein kinase, a universal regulator of mitosis, is controlled positively and negatively by phosphorylation, and by association with B-type mitotic cyclins. In addition, activation and inactivation of p34(cdc2) are induced by Ca2+ and prevented by Ca2+ chelators in permeabilized cells and cell-free systems. This suggests that intracellular Ca2+ transients may play an important physiological role in the control of p34(cdc2) kinase activity. We have found that activators of protein kinase C can be used to block cell cycle-related alterations in intracellular Ca2+ concentration ([Ca2+](i)) in early sea urchin embryos without altering the normal resting level of Ca2+. We have used this finding to investigate whether [Ca2+](i) transients control p34(cdc2) kinase activity in living cells via a mechanism that involves cyclin B or the phosphorylation state of p34(cdc2). In the present study we show that the elimination of [Ca2+](i), transients during interphase blocks p34(cdc2) activation and entry into mitosis, while the elimination of mitotic [Ca2+](i) transients prevents p34(cdc2) inactivation and exit from mitosis. Moreover, we find that [Ca2+](i) transients are not required for the synthesis of cyclin B, its binding to p34(cdc2) Or its destruction during anaphase. However, in the absence of interphase [Ca2+](i) transients p34(cdc2) does not undergo the tyrosine dephosphorylation that is required for activation, and in the absence of mitotic [Ca2+](i) transients p34(cdc2) does not undergo threonine dephosphorylation that is normally associated with inactivation. These results provide evidence that intracellular [Ca2+](i) transients trigger the dephosphorylation of p34(cdc2) at key regulatory sites, thereby controlling the timing of mitosis entry and exit. C1 Univ Stirling, Inst Biol Sci, Stirling FK9 4LA, Scotland. NICHHD, Lab Cellular & Mol Biophys, Bethesda, MD 20892 USA. Univ Newcastle Upon Tyne, Dept Physiol Sci, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. Worcester Fdn Biomed Res, Shrewsbury, MA 01545 USA. RP Whalley, T (reprint author), Univ Stirling, Inst Biol Sci, Stirling FK9 4LA, Scotland. OI Whalley, Tim/0000-0003-3362-0006 NR 56 TC 3 Z9 4 U1 0 U2 2 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 2000 VL 349 BP 489 EP 499 DI 10.1042/0264-6021:3490489 PN 2 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338LU UT WOS:000088422000013 PM 10880348 ER PT J AU Wellmann, A Thieblemont, C Pittaluga, S Sakai, A Jaffe, ES Siebert, P Raffeld, M AF Wellmann, A Thieblemont, C Pittaluga, S Sakai, A Jaffe, ES Siebert, P Raffeld, M TI Detection of differentially expressed genes in lymphomas using cDNA arrays: identification of clusterin as a new diagnostic marker for anaplastic large-cell lymphomas SO BLOOD LA English DT Article ID RETE TESTIS FLUID; TRANSCRIPTIONAL PROGRAM; KINASE ALK; HEAT-SHOCK; MICROARRAY; PATTERNS; DEATH; PROTEINS; CLASSIFICATION; RESISTANCE AB This study reports the first use of gene array technology for the identification of a tumor-specific marker in lymphoid neoplasms. The differential gene expression of 31 hematopoietic cell lines, representing most major lymphoma subgroups of B- and T-cell origin, was assessed by hybridizing labeled complementary DNA to Atlas human expression arrays containing 588 genes. Genes known to be specific for B, T, or myelomonocytic lineages were appropriately identified in the arrays, validating the general utility of this approach. One gene, clusterin, not previously known to be expressed in lymphoid neoplasms, was specifically found in all 4 anaplastic large-cell lymphoma (ALCL) cell lines, but not in any of the 27 remaining tumor lines. Using a monoclonal antibody against clusterin, its differential expression was confirmed by Western blotting and immunohistochemistry. A total of 198 primary lymphomas (representing most major lymphoma subtypes), including 36 cases of systemic ALCL, were surveyed for clusterin expression by immunohistochemistry and Western blotting. All of the 36 ALCL cases marked for clusterin, with most cases showing moderate to strong staining in the majority of neoplastic cells. Clusterin expression was not related to expression of anaplastic lymphoma kinase-1. With 2 exceptions, none of the remaining 162 non-ALCL cases marked with the clusterin antibody, including Hodgkin disease and primary cutaneous ALCL. In reactive lymphoid tissues, only follicular dendritic cells and fibroblastic reticular cells exhibited staining. Clusterin is a highly conserved glycoprotein implicated in intercellular and cell matrix interactions, regulation of the complement system, lipid transport, stress responses, and apoptosis, Although its function in ALCL is unknown, the unique expression of clusterin within this category of lymphoma provides an additional marker for the diagnosis of ALCL, This study illustrates the enormous potential of gene array technologies for diagnostic marker discovery. (C) 2000 by The American Society of Hematology. C1 NCI, Pathol Lab, Hematopathol Sect, NIH, Bethesda, MD 20892 USA. Clontech Labs, Palo Alto, CA USA. RP Raffeld, M (reprint author), NCI, Pathol Lab, Hematopathol Sect, NIH, Bldg 10,Rm 2N110,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 42 TC 139 Z9 143 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 2000 VL 96 IS 2 BP 398 EP 404 PG 7 WC Hematology SC Hematology GA 335GB UT WOS:000088234800004 PM 10887098 ER PT J AU Quintanilla-Martinez, L Kumar, S Fend, F Reyes, E Teruya-Feldstein, J Kingma, DW Sorbara, L Raffeld, M Straus, SE Jaffe, ES AF Quintanilla-Martinez, L Kumar, S Fend, F Reyes, E Teruya-Feldstein, J Kingma, DW Sorbara, L Raffeld, M Straus, SE Jaffe, ES TI Fulminant EBV+ T-cell lymphoproliferative disorder following acute/chronic EBV infection: a distinct clinicopathologic syndrome SO BLOOD LA English DT Article ID EPSTEIN-BARR-VIRUS; PROTEIN-1 ONCOGENE DELETIONS; NON-HODGKINS-LYMPHOMAS; REED-STERNBERG CELLS; HEMOPHAGOCYTIC SYNDROME; MONOCLONAL PROLIFERATION; GENE-EXPRESSION; DISEASE; MONONUCLEOSIS; ASSOCIATION AB This study describes the clinicopathologic features of 5 patients who developed a fulminant Epstein-Barr virus (EBV)-positive clonal T-cell lymphoproliferative disorder (LPD) after acute EBV infection. One additional patient developed a similar disorder in the setting of long-standing chronic active EBV infection. Detailed immunophenotyping, in situ hybridization for EBV early RNA-1 (EBER1) and polymerase chain reaction (PCR) analyses for immunoglobulin (Ig) heavy chain and T-cell receptor (TCR)-gamma gene rearrangements were performed on paraffin-embedded tissue from all patients. In addition, EBV strain typing and detection of the characteristic 30-bp deletion of the latent membrane protein-1 (LMP-1) gene were performed by PCR, Controls included 8 cases of uncomplicated infectious mononucleosis (IM). patients included 4 males and 2 females with a median age of 18 years (2-37 years). Three patients were Mexican, 2 were white, and 1 was of Asian descent. All presented with fever, hepatosplenomegaly, and pancytopenia; 5 were previously healthy, but had a clinical history of a recent viral-like upper respiratory illness (1 week to 2 months), and 1 patient had documented chronic active EBV infection for 7 years. Serologic data for EBV were incomplete but titers were either negative or only modestly elevated in 3 cases. In 1 case serology was consistent with severe chronic active EBV infection. In the remaining 2 cases serologic studies were not performed. All patients died within 7 days to 8 months of presentation with T-cell LPD. On histologic examination, the liver and spleen showed prominent sinusoidal and portal lymphoid infiltrates of CD3(+), beta F1(+), EBER1(+) T cells lacking significant cytologic atypia. Two cases were CD4(+), 2 cases were CD8(+), and 2 cases had admired CD4+ and CD8+ cells without clear subset predominance. All were TIA-1(+), CD56(-), Only rare B cells were noted. Marked erythrophagocytosis was present Molecular analysis revealed identical T-cell clones in 2 or more sites (liver, spleen, lymph node) in 5 cases. All patients carried type A EBV; 4 cases had wild-type EBV-LMP, and 2 showed the 30-bp deletion. This fulminant T-cell LPD after acute/chronic EBV infection is characterized by hepatosplenomegaly, often without significant lymphadenopathy, fever, liver failure, pancytopenia, and erythrophagocytosis indicative of a hemophagocytic syndrome, EBV serology may be misleading, with lack of elevated them, The presence of an EBER1(+) T-cell infiltrate with scant B cells should alert one to this diagnosis. Although cytologic atypia is minimal, studies for T-cell clonality confirm the diagnosis. (C) 2000 by The American Society of Hematology. C1 NCI, Pathol Lab, Hematopathol Sect, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Inst Nacl Nutr Salvador Zubiran, Dept Pathol, Mexico City, DF, Mexico. GSF, Res Ctr Environm & Hlth, Inst Pathol, Neuherberg, Germany. RP Jaffe, ES (reprint author), NCI, Pathol Lab, Hematopathol Sect, Bldg 10,Room 2N202,10 Ctr Dr,MSC 1500, Bethesda, MD 20892 USA. NR 59 TC 153 Z9 164 U1 0 U2 4 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 2000 VL 96 IS 2 BP 443 EP 451 PG 9 WC Hematology SC Hematology GA 335GB UT WOS:000088234800010 PM 10887104 ER PT J AU Walker, RE Bechtel, CM Natarajan, V Baseler, M Hege, KM Metcalf, JA Stevens, R Hazen, A Blaese, RM Chen, CC Leitman, SF Palensky, J Wittes, J Davey, RT Falloon, J Polis, MA Kovacs, JA Broad, DF Levine, BL Roberts, MR Masur, H Lane, HC AF Walker, RE Bechtel, CM Natarajan, V Baseler, M Hege, KM Metcalf, JA Stevens, R Hazen, A Blaese, RM Chen, CC Leitman, SF Palensky, J Wittes, J Davey, RT Falloon, J Polis, MA Kovacs, JA Broad, DF Levine, BL Roberts, MR Masur, H Lane, HC TI Long-term in vivo survival of receptor-modified syngeneic T cells in patients with human immunodeficiency virus infection SO BLOOD LA English DT Article ID CD28 COSTIMULATION; ADOPTIVE TRANSFER; LYMPHOCYTES-T; RNA LEVELS; IN-VIVO; VIREMIA; INFUSION; AIDS; TRANSDUCTION; INDIVIDUALS AB To study human immunodeficiency virus (HIV)-specific cellular immunity in vivo, we transferred syngeneic lymphocytes after ex vivo expansion and transduction with a chimeric receptor gene (CD4/CD3-zeta) between identical twins discordant for HIV infection. Single and multiple infusions of 10(10) genetically modified CD8(+) T cells resulted in peak fractions in the circulation of approximately 10(4) to 10(5) modified cells/10(6) mononuclear cells at 24 to 48 hours, followed by 2- to 3-log declines by 8 weeks. In an effort to provide longer high-level persistence of the transferred cells and possibly enhance anti-HIV activity, we administered a second series of infusions in which both CD4(+) and CD8(+) T cells were engineered to express the chimeric receptor and were costimulated ex vivo with beads coated with anti-CDS and anti-CD28. Sustained fractions of approximately 10(3) to 10(4) modified cells/10(6) total CD4(+) or CD8(+) cells persisted for at least 1 year. Assessment of in vivo trafficking of the transferred cells by lymphoid tissue biopsies revealed the presence of modified cells in proportions equivalent to or below those in the circulation. The cell infusions were well tolerated and were not associated with substantive Immunologic or virologic changes. Thus, adoptive transfer of genetically modified HIV-antigen-specific T cells was safe. Sustained survival in the circulation was achieved when modified CD4(+) and CD8(+) T cells were infused together after ex vivo costimulation, indicating the important role played by antigen-specific CD4(+) T cells in providing "help" to cytotoxic effecters. (C) 2000 by The American Society of Hematology. C1 NIH, Dept Nucl Med, Bethesda, MD 20892 USA. NIH, Dept Transfus Med, Bethesda, MD 20892 USA. NIH, Dept Crit Care Med, Bethesda, MD 20892 USA. NIAID, Clin Gene Therapy Branch, Natl Human Genome Res Inst, Clin & Mol Retrovirol Sect,Lab Immunoregulat, Bethesda, MD 20892 USA. SAIC, Frederick Canc Res & Dev Ctr, Frederick, MD USA. Cell Genesys Inc, Foster City, CA USA. Stat Collaborat, Washington, DC USA. Univ Penn, Ctr Canc, Leonard Madlyn Abramson Family Canc Res Inst, Philadelphia, PA 19104 USA. RP Lane, HC (reprint author), NIH, Dept Nucl Med, Bldg 10,Room 11S231,9000 Rockville, Bethesda, MD 20892 USA. RI Levine, Bruce/D-1688-2009; OI Polis, Michael/0000-0002-9151-2268 FU PHS HHS [N01-C0-56000] NR 36 TC 125 Z9 129 U1 1 U2 7 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 2000 VL 96 IS 2 BP 467 EP 474 PG 8 WC Hematology SC Hematology GA 335GB UT WOS:000088234800013 PM 10887107 ER PT J AU Paul, SP Taylor, LS Stansbury, EK McVicar, DW AF Paul, SP Taylor, LS Stansbury, EK McVicar, DW TI Myeloid specific human CD33 is an inhibitory receptor with differential ITIM function in recruiting the phosphatases SHP-1 and SHP-2 SO BLOOD LA English DT Article ID PROTEIN-TYROSINE-PHOSPHATASE; IMMUNOGLOBULIN SUPERFAMILY; B-CELL; ACID; ACTIVATION; BINDING; PHOSPHORYLATION; ANTIGEN; CD22; MOLECULES AB CD33 is a myeloid specific member of the sialic acid-binding receptor family and is expressed highly on myeloid progenitor cells but at much lower levels in differentiated cells. Human CD33 has two tyrosine residues in its cytoplasmic domain (Y340 and Y358), When phosphorylated, these tyrosines could function as docking sites for the phosphatases, SHP-1 and/or SHP-2, enabling CD33 to function as an inhibitory receptor. Here we demonstrate that CD33 is tyrosine phosphorylated in the presence of the phosphatase inhibitor, pervanadate, and recruits SHP-1 and SHP-2, Go-expression studies suggest that the Src-family kinase Lck is effective at phosphorylating Y340, but not Y358, suggesting that these residues may function in the selective recruitment of adapter molecules and have distinct functions. Further support for overlapping, but nonredundant, roles for Y340 and Y358 comes from peptide-binding studies that revealed the recruitment of both SHP-1 end SHP-2 to Y340 but only SHP-2 to Y358, Analysis using mutants of SHP-1 demonstrated that binding Y340 of CD33 was primarily to the amino Src homology-2 domain of SHP-1, The potential of CD33 to function as an inhibitory receptor was demonstrated by its ability to downregulate CD64-induced calcium mobilization in U937. The dependence of this inhibition on SHP-1 was demonstrated by blocking CD33-mediated effects with dominant negative SHP-1. This result implies that CD33 is an inhibitory receptor and also that SHP-1 phosphatase has a significant role in mediating CD33 function. Further studies are essential to identify the receptor(s) that CD33 inhibits in vivo and its function in myeloid lineage development. (C) 2000 by The American Society of Hematology. C1 NCI, Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Div Basic Sci, Frederick, MD 21702 USA. RP McVicar, DW (reprint author), NCI, Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Div Basic Sci, Bldg 560,Rm 31-93, Frederick, MD 21702 USA. RI McVicar, Daniel/G-1970-2015 NR 48 TC 101 Z9 102 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 2000 VL 96 IS 2 BP 483 EP 490 PG 8 WC Hematology SC Hematology GA 335GB UT WOS:000088234800015 PM 10887109 ER PT J AU Mackall, CL Stein, D Fleisher, TA Brown, MR Hakim, FT Bare, CV Leitman, SF Read, EJ Carter, CS Wexler, LH Gress, RE AF Mackall, CL Stein, D Fleisher, TA Brown, MR Hakim, FT Bare, CV Leitman, SF Read, EJ Carter, CS Wexler, LH Gress, RE TI Prolonged CD4 depletion after sequential autologous peripheral blood progenitor cell infusions in children and young adults SO BLOOD LA English DT Article ID SEVERE COMBINED IMMUNODEFICIENCY; BONE-MARROW TRANSPLANTATION; INTENSIVE CHEMOTHERAPY; IMMUNE RECONSTITUTION; STEM-CELLS; CANCER; AGE; THYMOPOIESIS; REGENERATION; RECOVERY AB Administration of mobilized peripheral blood progenitor cells (PBPCs) after high-dose chemotherapy rapidly restores multilineage hematopoiesis, but the ability of such products to restore lymphocyte populations remains unclear. In this report, we evaluated immune reconstitution in a series of patients treated with sequential cycles of high-dose chemotherapy, followed by autologous PBPC infusions (median CD34(+) cell dose 7.2 x 10(6) cells/kg [range 2-29.3]), Although patients experienced rapid reconstitution of B cells and CD8(+) T cells, we observed CD4 depletion and diminished immune responsiveness in all patients for several months after completion of therapy. Mature CD4(+) T cells contained within the grafts did not appear to contribute substantially to immune reconstitution because CD4 counts did not differ between recipients of unmanipulated T-cell replete Infusions versus CD34 selected, T-cell-depleted infusions. Rather, at 12 months after therapy, total CD4 count was inversely proportional to age (rho = -0.78, P = .04), but showed no relationship to CD34 cell dose (rho = -0.42, P = .26), suggesting that age-related changes within the host are largely responsible for the limited immune reconstitution observed. These results demonstrate that in the autologous setting, the infusion of large numbers of PBPCs is not sufficient to restore T-cell Immune competence and emphasize that specific approaches to enhance immune reconstitution are necessary if immune-based therapy is to be used to eradicate minimal residual disease after autologous PBPC transplantation. C1 NCI, Pediat Oncol Branch, Bethesda, MD 20892 USA. NCI, Med Branch, Bethesda, MD 20892 USA. NCI, Expt Immunol Branch, Bethesda, MD 20892 USA. NIH, Dept Transfus Med, Ctr Clin, Bethesda, MD 20892 USA. RP Mackall, CL (reprint author), 10 Ctr Dr,MSC 1928, Bethesda, MD 20892 USA. NR 33 TC 68 Z9 68 U1 0 U2 0 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 2000 VL 96 IS 2 BP 754 EP 762 PG 9 WC Hematology SC Hematology GA 335GB UT WOS:000088234800051 PM 10887145 ER PT J AU Hukku, B Mally, M Cher, ML Peehl, DM Kung, HF Rhim, JS AF Hukku, B Mally, M Cher, ML Peehl, DM Kung, HF Rhim, JS TI Stepwise genetic changes associated with progression of nontumorigenic HPV-18 immortalized human prostate cancer-derived cell line to a malignant phenotype SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID PAPILLOMAVIRUS TYPE-18 DNA; ALLELIC LOSS; DELETION; ABNORMALITIES; CARCINOMA; FREQUENCY; LOCI AB Cytogenetics, fluorescence in situ hybridization (FISH), and comparative genomic hybridization (CGH) were used to identify genes that are involved in the development and progression of prostate cancer. For that purpose, we chose a cell line established in vitro from a prostatic adenocarcinoma which was nontumorigenic in nude mice and followed its progression to a tumorigenic cell line. Stepwise changes were observed in the cell line as it became tumorigenic. The composite karyotype at the nontumorigenic stage (CA-HPV-10) was 68 similar to 77,XXY, -(1, 9, 13, 14, 19, 22),+(4, 5, 11, 18, 20, 21),+ (del(1) (q23q31)=M1 (two copies), + der(9)t(1;9)(q24 similar to q31;p23) = M5(two copies),der(14)t(14;?)(q10;?) = M17 in the majority of metaphases. These two derivative chromosomes were also observed a previous study. Our CGH analysis clearly showed that this deleted region in M1 is, in fact, translocated with derivative M5 and, in reality, is amplified. The cell line established from nodule (SCID 5029 p11, showed a number of new changes, as described; however, the most significant change was amplification of the 8q23 similar to qter region, harboring c-myc. This region was translocated with chromosomes 2, 4, and 16 as der(2) t(2;8)(q33;q23)=M12, der(4)t(4;8)(q34;q23)=M11, and der(16)t(8;16)(q24;q21)=M9. We deduce from our study that amplification of c-myc and other genes in the 8q23 similar to qter region were important in progression but did not lead to tumorigenicity. The population that became tumorigenic (SCID 5019 II) showed almost all of the same changes in the karyotype as observed in the nodular cell line; the only significant change was the appearance of der(11)t(4;11)(q32;q22)=M7 and the addition of another copy of t(3q;7p)=M2. These new changes lead to loss of chromosomes 3p, 4pter similar to q34, 6, 7q21 similar to qter, 11q22 similar to qter, and 18q, and gain of 3q, 7p, 8q23 similar to qter, and 11pter similar to q22, before the cell line became tumorigenic. The clonal selection of the population is proven by the presence of a number of the same derivative chromosomes in both the nodular and tumorigenic cell line. As it progressed to tumorigenicity, some of the same changes observed in the original study re-appear at different stages of malignancy, although it was absent in the nontumorigenic cell line. These are: der(16)t(8;1 6)(q24;q21)=M9 in the nodular cell line and der(11)t(4;11)(q32;q22)=M7 in the tumorigenic cell line. In our system, amplificafion of c-myc and other genes in der(2)t(2;8)(q33;q23) =M12, der(4) t(4;8)(q34;q23= M11 together with the presence of der(16)t(8;16)(q24;q21)=M9 and der(11)t(4;11)(q32;q22)=M5 makes the cell line tumorigenic. It is either nontumorigenic, with the presence of a marker equivalent to der(16)=M9 and der(11)=M7 observed in the original study, and only nodular (SCIL) 5019 p11, present study), with the presence of number of markers with c-myc amplification (M9, M11, and M12). There is accumulation of all the above-mentioned changes in the same cell before it becomes tumorigenic. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Childrens Hosp Michigan, Dept Pediat, Cell Culture Lab, Detroit, MI 48201 USA. Wayne State Univ, Sch Med, Detroit, MI USA. Wayne State Univ, Karamanos Canc Inst, Dept Urol & Pathol, Detroit, MI USA. Stanford Univ, Dept Urol, Sch Med, Stanford, CA 94305 USA. NCI, Lab Biochem Physiol, Frederick, MD 21701 USA. RP Hukku, B (reprint author), Childrens Hosp Michigan, Dept Cell Culture, 3901 Beaubien Blvd, Detroit, MI 48201 USA. FU NCI NIH HHS [N0I-CB33063] NR 23 TC 10 Z9 12 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0165-4608 J9 CANCER GENET CYTOGEN JI Cancer Genet. Cytogenet. PD JUL 15 PY 2000 VL 120 IS 2 BP 117 EP 126 DI 10.1016/S0165-4608(00)00216-8 PG 10 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 347JF UT WOS:000088925000005 PM 10942801 ER PT J AU Marrogi, A Pass, HI Khan, M Metheny-Barlow, LJ Harris, CC Gerwin, BI AF Marrogi, A Pass, HI Khan, M Metheny-Barlow, LJ Harris, CC Gerwin, BI TI Human mesothelioma samples overexpress both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (NOS2): In vitro antiproliferative effects of a COX-2 inhibitor SO CANCER RESEARCH LA English DT Article ID NONSTEROIDAL ANTIINFLAMMATORY DRUGS; APOPTOTIC CELL-DEATH; NF-KAPPA-B; SELECTIVE-INHIBITION; IN-VIVO; EXPRESSION; LINES; MECHANISMS; INDUCTION; RAT AB Accumulating data demonstrate overexpression of both inducible NO synthase (NOS2) and cyclooxygenase-2 (COX2) in many epithelial neoplasias, In addition, cyclooxygenase inhibitors have been shown to have antineoplastic and prophylactic efficacy against human colon cancer and in mouse models of this disease. Mesothelioma arises in a context of asbestos exposure and chronic inflammation, which would be expected to enhance the expression of these inducible enzymes. This study demonstrates that both inducible enzymes were expressed in 30 human mesothelioma tissues but were not detectable in nonreactive mesothelial tissues from the same individuals. In contrast, areas of reactive mesothelial cells stained positively for these enzymes. In vitro exposure of human mesothelioma cell lines to the COX2 inhibitor, NS398, revealed dose- and time-dependent antiproliferative activity, whereas the NOS2 inhibitor, 1400W, had no detectable inhibitory effect, Surprisingly, nonmalignant human mesothelial isolates expressed both NOS2 and COX2 in vitro at the same level as mesothelioma cell lines but were less sensitive to NS398 inhibition. This finding indicates that these nonmalignant isolates may retain properties of reactive mesothelial cells and suggests that targets in addition to COX2 mag be involved in the antiproliferative response of mesothelioma cell lines. These results have clinical significance because of the selective activity of the drug coupled with the therapeutic resistance and poor prognosis of mesothelioma, The findings presented here suggest that further preclinical studies of these inhibitors in animal models of mesothelioma would be of great interest. C1 NCI, Human Carcinogenesis Lab, Bethesda, MD 20892 USA. Wayne State Univ, Karmanos Canc Inst, Thorac Oncol Program, Detroit, MI 48201 USA. RP Gerwin, BI (reprint author), NCI, Human Carcinogenesis Lab, Bldg 37,Room 2C07, Bethesda, MD 20892 USA. RI Metheny-Barlow, Linda/C-3123-2011 NR 34 TC 81 Z9 89 U1 2 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 2000 VL 60 IS 14 BP 3696 EP 3700 PG 5 WC Oncology SC Oncology GA 337ML UT WOS:000088365300003 PM 10919635 ER PT J AU Noonan, FP Otsuka, T Bang, S Anver, MR Merlino, G AF Noonan, FP Otsuka, T Bang, S Anver, MR Merlino, G TI Accelerated ultraviolet radiation-induced carcinogenesis in hepatocyte growth factor/scatter factor transgenic mice SO CANCER RESEARCH LA English DT Article ID SQUAMOUS-CELL CARCINOMA; FACTOR SCATTER FACTOR; MALIGNANT-MELANOMA; C-MET; SUN EXPOSURE; SKIN-CANCER; GENOTOXIC STRESS; PATHOGENESIS; EXPRESSION; INDUCTION AB The dramatic rise in incidence of malignant melanoma experienced by populations both within the United States and throughout the world over the last several decades has been attributed to enhanced exposure to the UV spectrum of sunlight radiation. This hypothesis can now be tested using genetically engineered mouse models predisposed to malignant melanoma, Here we use melanoma-prone transgenic mice inappropriately expressing hepatocyte growth factor/scatter factor (HGF/SF) in the skin as an experimental model system to ascertain the consequences of a chronic regimen of suberythemal UV radiation on melanoma genesis. HGF/SF is a multifunctional regulator capable of stimulating growth, motility, invasiveness, and morphogenetic transformation in cells, including melanocytes, expressing its receptor tyrosine kinase Met. HGF/SF transgenic mice demonstrate ectopic interfollicular localization and accumulation of melanocytes within the truncal dermis, epidermis, and junction and if untreated develop primary cutaneous melanoma with a mean onset age of similar to 21 months. Transgenic mice and their wild-type littermates subjected to UV radiation three times weekly using FS40 sunlamps (60% UVB and 40% UVA), with daily UV doses graded from 2.25 to 6.0 KJ/m(2), developed skin tumors with a mean onset age of 26 and 37 weeks, respectively (P < 0.001, Kaplan-Meier log rank test), However, the repeated doses of suberythemal UV radiation used in this study failed to accelerate melanoma genesis, instead inducing the development of nonmelanoma tumors that included squamous cell carcinomas, squamous papillomas, and sarcomas, The conspicuous absence of melanocytic tumors occurred despite the immunohistochemical detection of a significant stimulation (P < 0.001) in melanocyte-specific bromodeoxyuridine incorporation in response to only 2 weeks of UV irradiation (total UV dose of 13.5 kJ/m(2)), resulting in 2.6- and 4.6-fold increases in the number of melanocytes in the dermis and epidermis, respectively. These data indicate that chronic suberythemal UV radiation preferentially favors the development of nonmelanocytic over melanocytic neoplasms in this transgenic animal, consistent with the pathogenesis proposed for sun exposure-associated skin cancer based on retrospective studies in the human population. Our findings suggest that the HGF/SF transgenic mouse will be useful as an experimental model for determining the consequences of exposure to various regimens of UV radiation and for elucidating the mechanisms by which such consequences are realized. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. George Washington Univ, Med Ctr, Dept Dermatol, Lab Photobiol & Photoimmunol, Washington, DC 20037 USA. NCI, Pathol Histotechnol Lab, Sci Applicat Int Corp, Frederick Canc Res & Dev Ctr, Ft Detrick, MD 21702 USA. RP Merlino, G (reprint author), NCI, Mol Biol Lab, NIH, Bldg 37,Room 2E24, Bethesda, MD 20892 USA. FU NCI NIH HHS [N0I-CO-56000] NR 51 TC 67 Z9 68 U1 0 U2 6 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 2000 VL 60 IS 14 BP 3738 EP 3743 PG 6 WC Oncology SC Oncology GA 337ML UT WOS:000088365300011 PM 10919643 ER PT J AU Sinha, R Kulldorff, M Swanson, CA Curtin, J Brownson, RC Alavanja, MCR AF Sinha, R Kulldorff, M Swanson, CA Curtin, J Brownson, RC Alavanja, MCR TI Dietary heterocyclic amines and the risk of lung cancer among Missouri women SO CANCER RESEARCH LA English DT Article ID UNITED-STATES; VARYING DEGREES; FAT; CHOLESTEROL; MORTALITY; DONENESS; HEALTH; FOODS; MEAT; MEN AB Heterocyclic amines (HCAs) such as 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) are found in meats cooked at high temperatures. In rodents, MeIQx induces lung turners. The purpose of this study was to investigate lung cancer risk posed by different HCAs in the diet. A population-based case-control study of 593 cases and 623 frequency-matched controls including both nonsmoking and smoking women was conducted in Missouri, An administered food frequency questionnaire with detailed questions on meat consumption, degrees of internal doneness, surface browning/charring, and cooking technique was linked to a database that provided exposure estimates of three HCAs. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression. When comparing the 90(th) and 10(th) percentiles, significant excess risks were observed for MeIQx (OR, 1.5; CI, 1.1-2.0), but not fur DiMeIQx (OR, 1.2; CI, 0.9-1.6) or PhIP (OR, 0.9; CI, 0.8-1.1). MeIQx consumption was associated with increased risk of lung cancer for nonsmokers (OR, 3.6; CI, 1.3-10.3) and light/moderate smokers (OR, 2.1; CI, 1.3-3.3), hut not for heavy smokers (OR, 1.0; CI, 0.7-1.5), There was elevated risk with MeIQx: intake for subjects with squamous cell carcinomas (OR, 1.9; CI, 1.2-3.1) and "other histological cell types" (OR, 1.6; CI, 1.1-2.5), but not for subjects with small cell carcinomas and adenocarcinomas. Neither DiMeIQx: nor PhIP showed an association with smoking categories or lung cancer histology, In conclusion, MeIQx may be associated with lung cancer risk, but DiMeIQx and PhIP are probably not associated with lung canter risk. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ Connecticut, Sch Med, Dept Community Med & Hlth Care, Div Biostat, Farmington, CT 06030 USA. Informat Management Serv Inc, Silver Spring, MD 20904 USA. St Louis Univ, Sch Publ Hlth, Dept Community Hlth, St Louis, MO 63103 USA. St Louis Univ, Sch Publ Hlth, Prevent Res Ctr, St Louis, MO 63103 USA. RP Sinha, R (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS Room 7028, Bethesda, MD 20892 USA. RI Kulldorff, Martin/H-4282-2011; Sinha, Rashmi/G-7446-2015; OI Sinha, Rashmi/0000-0002-2466-7462; Kulldorff, Martin/0000-0002-5284-2993 NR 29 TC 50 Z9 50 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 2000 VL 60 IS 14 BP 3753 EP 3756 PG 4 WC Oncology SC Oncology GA 337ML UT WOS:000088365300014 PM 10919646 ER PT J AU Li, WQ Hyun, T Heller, M Yam, A Flechner, L Pierce, JH Rudikoff, S AF Li, WQ Hyun, T Heller, M Yam, A Flechner, L Pierce, JH Rudikoff, S TI Activation of insulin-like growth factor I receptor signaling pathway is critical for mouse plasma cell tumor growth SO CANCER RESEARCH LA English DT Article ID KINASE-C-DELTA; MULTIPLE-MYELOMA; MURINE PLASMACYTOMAGENESIS; TYROSINE KINASE; V-MYC; MICE; TRANSLOCATION; EXPRESSION; GENE; ABL AB Plasma cell neoplasia in humans generally occurs as multiple myeloma, an incurable form of cancer. Tumors with marked similarity can be induced in mice hg a variety of agents, including chemicals, silicone, and oncogene-containing retroviruses, suggesting the use of murine tumors as an informative model to study plasma cell disease. Herein, we have focused on the role of insulin-like growth factor I receptor (IGF-IR) signaling in the development of plasma cell disease. The insulin receptor substrate 2/phosphatidylinositol 3'-kinase/p70S6K pathway was found to be either constitutively or IGF-I-dependently activated in all plasma cell tumors. Biological relevance was demonstrated in that plasma cell lines with up-regulated IGF IR expression levels exhibited mitogenic responses to IGF-I, More importantly, expression of a dominant-negative mutant of IGF-IR in these lines strongly suppressed tumorigenesis in vivo. Taken together, those results demonstrate that up-regulation and activation of IGF-IR and the downstream signaling pathway involving insulin receptor substrate 2, phosphatidylinositol 3'-kinase, and p70S6K may play an important role in the development of a broad spectrum of plasma cell tumors. C1 Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Dept Oncol,NRB, Washington, DC 20007 USA. NCI, Cellular & Mol Biol Lab, Bethesda, MD 20892 USA. RP Li, WQ (reprint author), Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Dept Oncol,NRB, E407,3970 Reservoir Rd NW, Washington, DC 20007 USA. NR 39 TC 23 Z9 24 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 2000 VL 60 IS 14 BP 3909 EP 3915 PG 7 WC Oncology SC Oncology GA 337ML UT WOS:000088365300036 PM 10919668 ER PT J AU Carson, DD Bagchi, I Dey, SK Enders, AC Fazleabas, AT Lessey, BA Yoshinaga, K AF Carson, DD Bagchi, I Dey, SK Enders, AC Fazleabas, AT Lessey, BA Yoshinaga, K TI Embryo implantation SO DEVELOPMENTAL BIOLOGY LA English DT Review ID LEUKEMIA INHIBITORY FACTOR; UTERINE EPITHELIAL-CELLS; COLONY-STIMULATING FACTOR; ACTIVATED RECEPTOR-ALPHA; GROWTH-FACTOR RECEPTOR; PERIIMPLANTATION MOUSE UTERUS; CALCITONIN-GENE EXPRESSION; EXTRACELLULAR-MATRIX COMPONENTS; HEPARAN-SULFATE PROTEOGLYCANS; BABOON PAPIO-ANUBIS C1 Univ Delaware, Dept Biol Sci, Newark, DE 19716 USA. Rockefeller Univ, Populat Council, New York, NY 10021 USA. Univ Kansas, Med Ctr, Dept Mol & Integrat Physiol, Kansas City, KS 66160 USA. Univ Calif Davis, Dept Cell Biol & Human Anat, Davis, CA 95616 USA. Univ Illinois, Dept Obstet & Gynecol, Chicago, IL 60612 USA. Univ N Carolina, Dept Obstet & Gynecol, Div Reprod Endocrinol & Infertil, Chapel Hill, NC 27599 USA. NICHD, Reprod Sci Branch, Populat Res Ctr, NIH, Bethesda, MD 20892 USA. RP Carson, DD (reprint author), Univ Delaware, Dept Biol Sci, Newark, DE 19716 USA. FU NICHD NIH HHS [HD29968, HD29963, HD34760] NR 228 TC 470 Z9 502 U1 2 U2 22 PU ACADEMIC PRESS INC 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 2000 VL 223 IS 2 BP 217 EP 237 DI 10.1006/dbio.2000.9767 PG 21 WC Developmental Biology SC Developmental Biology GA 337RX UT WOS:000088376400001 PM 10882512 ER PT J AU Dichoso, D Brodigan, T Chwoe, KY Lee, JS Llacer, R Park, M Corsi, AK Kostas, SA Fire, A Ahnn, J Krause, M AF Dichoso, D Brodigan, T Chwoe, KY Lee, JS Llacer, R Park, M Corsi, AK Kostas, SA Fire, A Ahnn, J Krause, M TI The MADS-Box factor CeMEF2 is not essential for Caenorhabditis elegans myogenesis and development SO DEVELOPMENTAL BIOLOGY LA English DT Article DE MEF2; MADS box; myogenesis; C. elegans; SRF ID SERUM RESPONSE FACTOR; D-MEF2 TRANSCRIPTION FACTOR; MUSCLE-SPECIFIC GENES; C-ELEGANS; DROSOPHILA MEF2; MOUSE EMBRYOGENESIS; EXPRESSION; ENHANCER; PROTEIN; CELLS AB MEF2 is an evolutionarily conserved MADS (MCM1, Agamous, Deficiens, and serum response factor) box-type transcription factor that plays a critical role in vertebrate and Drosophila melanogaster myogenesis. We have addressed the developmental role of the single MEF2-like factor, CeMEF2, in Caenorhabditis elegans. Using expression assays and two mef-2 deletion alleles, we show that CeMEF2 is not required for proper myogenesis or development. Moreover, a putative null mef-2 allele fails to enhance or suppress the phenotypes of mutants in CeMyoD or CeTwist. Our results suggest that despite its evolutionary conservation of sequence and DNA binding properties, CeMEF2 has adopted a divergent role in development in the nematode compared with Drosophila and vertebrates. C1 NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Kwangju Inst Sci & Technol, Dept Life Sci, Kwangju, South Korea. Carnegie Inst Sci, Dept Embryol, Baltimore, MD 21210 USA. RP NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. FU NIGMS NIH HHS [R01-GM37706, T32 GM07231, R01 GM037706] NR 64 TC 27 Z9 33 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 EI 1095-564X J9 DEV BIOL JI Dev. Biol. PD JUL 15 PY 2000 VL 223 IS 2 BP 431 EP 440 DI 10.1006/dbio.2000.9758 PG 10 WC Developmental Biology SC Developmental Biology GA 337RX UT WOS:000088376400016 PM 10882527 ER PT J AU Kramer, PR Wray, S AF Kramer, PR Wray, S TI Novel gene expressed in nasal region influences outgrowth of olfactory axons and migration of luteinizing hormone-releasing hormone (LHRH) neurons SO GENES & DEVELOPMENT LA English DT Article DE olfactory; migration; LHRH; GnRH ID CELL-ADHESION MOLECULE; EXPLANT CULTURES; GLOMERULAR FORMATION; DEVELOPING MOUSE; DEVELOPING RAT; MESSENGER-RNA; GUIDANCE; BULB; RECEPTORS; NERVE AB Although a variety of cues have been implicated in axonal targeting during embryogenesis and regeneration, the precise mechanisms guiding olfactory axons remain unclear. Appropriate olfactory axon pathfinding is essential for functional chemoreceptive and pheromone receptive systems. Olfactory axon pathfinding is also necessary for establishment of the neuroendocrine LHRH system, cells critical for reproductive function. LHRH cells exhibit neurophilic migration moving from the nasal region along olfactory axons into the brain. Factors involved in the migration of these neuroendocrine cells are as yet unresolved. We report identification of a novel factor termed nasal embryonic LHRH factor (NELF) that was discovered in a differential screen of migrating versus nonmigrating primary LHRH neurons. NELF is expressed in PNS and CNS tissues during embryonic development, including olfactory sensory cells and LHRH cells. NELF antisense experiments indicate that a reduction in NELF expression decreases olfactory axon outgrowth and the number of LHRH neurons that migrate out of the nasal tissue. These results demonstrate that NELF plays a role as a common guidance molecule for olfactory axon projections and subsequently, either directly or indirectly, in the neurophilic migration of LHRH cells. C1 NINDS, Cellular & Dev Neurobiol Sect, NIH, Bethesda, MD 20892 USA. RP Wray, S (reprint author), NINDS, Cellular & Dev Neurobiol Sect, NIH, Bethesda, MD 20892 USA. OI Kramer, Phillip/0000-0003-0117-542X; wray, susan/0000-0001-7670-3915 NR 46 TC 88 Z9 91 U1 0 U2 1 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 0890-9369 J9 GENE DEV JI Genes Dev. PD JUL 15 PY 2000 VL 14 IS 14 BP 1824 EP 1834 PG 11 WC Cell Biology; Developmental Biology; Genetics & Heredity SC Cell Biology; Developmental Biology; Genetics & Heredity GA 335VJ UT WOS:000088265200012 PM 10898796 ER PT J AU Kim, SC Dougherty, ER Chen, YD Sivakumar, K Meltzer, P Trent, JM Bittner, M AF Kim, SC Dougherty, ER Chen, YD Sivakumar, K Meltzer, P Trent, JM Bittner, M TI Multivariate measurement of gene expression relationships SO GENOMICS LA English DT Article ID MICROARRAY HYBRIDIZATION; SACCHAROMYCES-CEREVISIAE; CDNA MICROARRAY; PATTERNS; CELLS AB The operational activities of cells are based on an awareness of their current state, coupled to a programmed response to internal and external cues in a context-dependent manner. One key goal of functional genomics is to develop analytical methods for delineating the ways in which the individual actions of genes are integrated into our understanding of the increasingly complex systems of organelle, cell, organ, and organism. This paper describes a novel approach to assess the codetermination of gene transcriptional states based upon statistical evaluation of reliably informative subsets of data derived from large-scale simultaneous gene expression measurements with cDNA microarrays. The method finds associations between the expression patterns of individual genes by determining whether knowledge of the transcriptional levels of a small gene set can be used to predict the associated transcriptional state of another gene. To test this approach for identification of the relevant contextual elements of cellular response, we have modeled our approach using data from known gene response pathways including ionizing radiation and downstream targets of inactivating gene mutations. This approach strongly suggests that evaluation of the transcriptional status of a given gene(s) can be combined with data from global expression analyses to predict the expression level of another gene. With data sets of the size currently available, this approach should be useful in finding sets of genes that participate in particular biological processes. As larger data sets and more computing power become available, the method can be extended to validating and ultimately identifying biologic (transcriptional) pathways based upon large-scale gene expression analysis. (C) 2000 Academic Press. C1 Natl Human Genome Res Inst, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. Texas A&M Univ, Dept Elect Engn, College Stn, TX 77843 USA. Washington State Univ, Dept Elect Engn, Pullman, WA 99164 USA. RP Bittner, M (reprint author), Natl Human Genome Res Inst, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. NR 28 TC 98 Z9 100 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD JUL 15 PY 2000 VL 67 IS 2 BP 201 EP 209 DI 10.1006/geno.2000.6241 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 339JQ UT WOS:000088474300010 PM 10903845 ER PT J AU Gao, YT Shu, XO Dai, Q Potter, JD Brinton, LA Wen, WQ Sellers, TA Kushi, LH Ruan, ZX Bostick, RM Jin, F Zheng, W AF Gao, YT Shu, XO Dai, Q Potter, JD Brinton, LA Wen, WQ Sellers, TA Kushi, LH Ruan, ZX Bostick, RM Jin, F Zheng, W TI Association of menstrual and reproductive factors with breast cancer risk: Results from the Shanghai Breast Cancer Study SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID WOMEN; LACTATION; METAANALYSIS; EXPERIENCE; ABORTION; HISTORY AB The incidence of breast cancer among women in Shanghai, a traditionally low-risk population, has increased substantially over the past 20 years. To evaluate the association of menstrual and reproductive factors with breast cancer risk and the influence of these factors on the temporal trend of breast cancer incidence, we analyzed data from the Shanghai Breast Cancer Study, a population-based case-control study of breast cancer recently completed among Chinese women in urban Shanghai, In-person interviews were completed for 1,459 women newly diagnosed with breast cancer between ages 25 and 64 and for 1,556 controls frequency-matched to cases by age. Unconditional logistic regression was employed to estimate adjusted odds ratios (ORs) and 95% confidence intervals (CIs) related to menstrual and reproductive factors. Earlier menarcheal age, nulliparity, and later age at first live birth were associated with increased risk of breast cancer among both pre- and post-menopausal women, while never having breast-fed and later age at menopause were associated with elevated risk only among post-menopausal women. Among controls, 32% of younger women (less than or equal to 40 years) and 24% of older women (>40 years) reported starting menarche at age of 13 or younger, and this factor contributed to 44% of cases diagnosed among younger women and 26% to 28% of cases in older women. Older age at first live birth or at menopause explained a considerable portion of cases diagnosed in older, but not younger, women. Our study suggests that the changes in menstrual and reproductive patterns among women in Shanghai have contributed to the recent increase in breast cancer incidence, particularly among younger women. Int. J. Cancer 87:295-300, 2000. (C) 2000 Wiley-Liss, Inc. C1 Univ S Carolina, Dept Pediat, Columbia, SC 29203 USA. S Carolina Canc Ctr, Columbia, SC USA. Shanghai Canc Inst, Dept Epidemiol, Shanghai, Peoples R China. Univ S Carolina, Dept Epidemiol, Columbia, SC 29203 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. NCI, Environm Epidemiol Branch, Bethesda, MD 20892 USA. Mato Clin, Teachers Coll, Dept Hlth & Behav Studies, Rochester, MN USA. RP Shu, XO (reprint author), Univ S Carolina, Dept Pediat, 15 Richland Med Pk,Suite 301, Columbia, SC 29203 USA. RI Brinton, Louise/G-7486-2015; OI Brinton, Louise/0000-0003-3853-8562; Potter, John/0000-0001-5439-1500; Kushi, Lawrence/0000-0001-9136-1175 FU NCI NIH HHS [R01CA64277] NR 23 TC 182 Z9 191 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 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD JUL 15 PY 2000 VL 87 IS 2 BP 295 EP 300 DI 10.1002/1097-0215(20000715)87:2<295::AID-IJC23>3.0.CO;2-7 PG 6 WC Oncology SC Oncology GA 329YZ UT WOS:000087937200023 PM 10861490 ER PT J AU Coleman, CN Govern, FS Svensson, G Mitchell, R Chaffey, JT AF Coleman, CN Govern, FS Svensson, G Mitchell, R Chaffey, JT TI The Harvard Joint Center for Radiation Therapy, 1968-1999: A unique concept and its relationship to the prevailing times in academic medicine SO INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS LA English DT Article DE radiation oncology; healthcare delivery; joint ventures; oncology; academic medicine; community science; collaboration ID ONCOLOGY; CANCER AB Purpose: Institutional structure, function, and philosophy reflect the organizational needs, and tend to mirror societal values of the times. For many gears, the field of radiation oncology had among its major academic centers, an organization that served as a model for collaboration among health care institutions in an effort to serve the common good of its patients, hospitals, professional colleagues, and community. For over three decades, the Joint Center for Radiation Therapy (JCRT) was a leader in developing new organizational approaches for academic and clinical radiation oncology through the philosophy of collaboration in patient care, education, and research. Methods and Results: In tracing the development and changes in organizational philosophy and structure of the JCRT, one can see the impact on academic oncology and cancer care through the emergence of both radiation and medical oncology as independent subspecialties, the importance of the National Cancer Act of 1971 accompanied by the growth of the NIH research and training programs and, more recently, the effect of the changing attitudes and approaches of hospitals, academicians, practitioners, and policy makers to health care delivery, structures, and cooperation. Conclusion: Lessons learned from the 31-year history of the JCRT may help provide organizational insight useful in guiding academic oncology and academic medical centers through periods of change, (C) 2000 Elsevier Science Inc. C1 Harvard Univ, Sch Med, Joint Ctr Radiat Therapy, Boston, MA 02115 USA. RP Coleman, CN (reprint author), NCI, Radiat Oncol Sci Program, Radiat Oncol Branch, NIH, Bldg 10,B3B69, Bethesda, MD 20892 USA. NR 10 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0360-3016 J9 INT J RADIAT ONCOL JI Int. J. Radiat. Oncol. Biol. Phys. PD JUL 15 PY 2000 VL 47 IS 5 BP 1357 EP 1369 DI 10.1016/S0360-3016(00)00548-4 PG 13 WC Oncology; Radiology, Nuclear Medicine & Medical Imaging SC Oncology; Radiology, Nuclear Medicine & Medical Imaging GA 334TW UT WOS:000088202600024 PM 10889391 ER PT J AU De Cuyper, M Bulte, JWM AF De Cuyper, M Bulte, JWM TI Preparation and characterization of a phospholipid membrane-bound tetrapeptide that corresponds to the C-terminus of the gastrin/cholecystokinin hormone family SO JOURNAL OF COLLOID AND INTERFACE SCIENCE LA English DT Article ID TRYPTOPHAN OCTYL ESTER; N-HYDROXYSULFOSUCCINIMIDE; POLY(ETHYLENE GLYCOL); LIPID BILAYERS; FLUORESCENCE; CARBODIIMIDE; CHOLECYSTOKININ; SURFACE; PROTEIN; RECEPTORS AB The present work deals with the synthesis of a hydrophobized peptide and its localization at the membrane surface, after its incorporation into phospholipid vesicles. The tetrapeptide, Trp-Met-Asp-Phe-NH2, which corresponds to the C-terminus of the cholecystokinin/gastrin hormone family, is conjugated to N-glutaryldioleoylphosphatidylethanolamine using a carwbodiimide-catalyzed reaction method, Sonication of the lipophilized hormone in the presence of dimyristoylphosphatidylcholine results in a strong sequestration of the conjugate in the artificial membrane structures that are formed, More detailed information on the localization of the peptide moiety with respect to the membrane surface is gathered from fluorescence measurements. Both the observed blue shift in the fluorescence spectra and the quenching of Trp emission in the presence of potassium iodide point to a partial screening of the hormone moiety from the surrounding aqueous phase. The different parameters that may influence the physicochemical behavior of a hydrophobized peptide in a membrane structure are briefly discussed. (C) 2000 Academic Press. C1 Katholieke Univ Leuven, Interdisciplinary Res Ctr, B-8500 Kortrijk, Belgium. NIH, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. RP De Cuyper, M (reprint author), Katholieke Univ Leuven, Interdisciplinary Res Ctr, Campus Kortrijk, B-8500 Kortrijk, Belgium. RI Bulte, Jeff/A-3240-2008 OI Bulte, Jeff/0000-0003-1202-1610 NR 57 TC 4 Z9 4 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0021-9797 J9 J COLLOID INTERF SCI JI J. Colloid Interface Sci. PD JUL 15 PY 2000 VL 227 IS 2 BP 421 EP 426 DI 10.1006/jcis.2000.6902 PG 6 WC Chemistry, Physical SC Chemistry GA 333JA UT WOS:000088126100021 ER PT J AU Yap, G Pesin, M Sher, A AF Yap, G Pesin, M Sher, A TI Cutting edge: IL-12 is required for the maintenance of IFN-gamma production in T cells mediating chronic resistance to the intracellular pathogen, Toxoplasma gondii SO JOURNAL OF IMMUNOLOGY LA English DT Article ID EXPERIMENTAL COLITIS; RECEPTOR REPERTOIRE; IMMUNE-RESPONSE; ACUTE INFECTION; TNF-ALPHA; MICE; TH1; EXPRESSION; ABSENCE; INDUCE AB IL-12 is required for the development of IFN-gamma-dependent resistance to intracellular pathogens but is not thought to play a major role in its maintenance, To directly assess the requirement for continuous IL-12 signaling in long-term cell-mediated immunity, recombinant cytokine was transiently administered to IL-12 p40-deficient mice during the first 2 wk of infection with the intracellular pathogen Toxoplasma gondii, As expected, these animals survived the acute phase and established chronic infections. However, 4-6 wk after IL-12 withdrawal, the mice exhibited increased brain cyst burdens and succumbed to toxoplasmic encephalitis, Reactivation was associated with a loss of T-dependent IFN-gamma production without a concomitant increase in Th2 cytokine expression. Importantly, parasite Ag-induced IFN-gamma synthesis by purified T cells from these animals could be restored by in vitro exposure to IL-12, These results argue that endogenous IL-12 is required for the long-term maintenance of IFN-gamma-dependent resistance against intracellular pathogens. C1 NIAID, Immunobiol Sect, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Sher, A (reprint author), NIAID, Immunobiol Sect, Parasit Dis Lab, NIH, Bldg 4,Room 126,Ctr Dr, Bethesda, MD 20892 USA. NR 23 TC 178 Z9 182 U1 0 U2 3 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 2000 VL 165 IS 2 BP 628 EP 631 PG 4 WC Immunology SC Immunology GA 332QU UT WOS:000088085600004 PM 10878333 ER PT J AU Kitani, A Chua, K Nakamura, K Strober, W AF Kitani, A Chua, K Nakamura, K Strober, W TI Activated self-MHC-reactive T cells have the cytokine phenotype of Th3/T regulatory cell 1 T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID GROWTH-FACTOR-BETA; POKEWEED MITOGEN; ORAL TOLERANCE; CD40 LIGAND; HELPER; MICE; AUTOIMMUNITY; EXPRESSION; INDUCTION; ANTIGEN AB In the present study, we show that human self-MHC-reactive (autoreactive) T cell clones are functionally distinct from Ag-specific T cell clones, Self-MHC-reactive T cells exhibited helper function for B cell Ig production when cultured with non-T cells alone, and they exhibit suppressor function when cultured with PWM- or rCD40 ligand (rCD40L)-activated non-T cells, whereas tetanus toroid (TT)-specific clones exhibited only helper function in the presence of TT with or without PWM or rCD40L, Addition of neutralizing Abs to the cultures showed that the suppression was mediated by TGF-beta but not by IL-10 or IFN-gamma, The self-MHC-reactive clones also inhibited proliferation of primary CD4(+) T cells and TT-specific T cell clones, but in this case the inhibition was mediated by both IL-10 and TGF-beta, In further studies, the interactions between self-MHC-reactive T cell clones and non-T cells that led to suppressor cytokine production have been explored. We found that prestimulation of non-T cells for 8 h with PWM or for 48 h for rCD40L results in non-T cells capable of inducing self-MHC-reactive T cell to produce high levels of TGF-beta and IL-10, In addition, these prestimulation times coincided with peak induction of HLA-DR and costimulatory B7 molecule (especially CD86) expression on B cells, Finally, addition of CTLA-4/Fc or blocking F(ab')(2) anti-CTLA-4 mAb, plus optimally stimulated non-T cells, to cultures of self-MHC-reactive clones inhibited the induction of TGF-beta but not IL-10 or IFN-gamma production. In summary, these studies show that activated self-MHC-reactive T cells have the cytokine phenotype of Th3 or T regulatory cell 1 and thus may be important regulatory cells that mediate oral and peripheral tolerance and prevent the development of autoimmunity. C1 NIAID, Mucosal Immun Sect, Clin Invest Lab, NIH, Rockville, MD 20852 USA. RP Strober, W (reprint author), NIAID, Mucosal Immun Sect, Clin Invest Lab, NIH, Bldg 10,11N238, Rockville, MD 20852 USA. NR 44 TC 95 Z9 104 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 2000 VL 165 IS 2 BP 691 EP 702 PG 12 WC Immunology SC Immunology GA 332QU UT WOS:000088085600012 PM 10878341 ER PT J AU Gurunathan, S Stobie, L Prussin, C Sacks, DL Glaichenhaus, N Fowell, DJ Locksley, RM Chang, JT Wu, CY Seder, RA AF Gurunathan, S Stobie, L Prussin, C Sacks, DL Glaichenhaus, N Fowell, DJ Locksley, RM Chang, JT Wu, CY Seder, RA TI Requirements for the maintenance of Th1 immunity in vivo following DNA vaccination: A potential immunoregulatory role for CD8(+) T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID LEISHMANIA-MAJOR INFECTION; MOUSE IL-12 RECEPTOR; INTERFERON-GAMMA; CUTANEOUS LEISHMANIASIS; PROTECTIVE IMMUNITY; BALB/C MICE; INTERLEUKIN-12; EXPRESSION; RESISTANCE; PARASITE AB Protective immunity against Leishmania major generated by DNA encoding the LACK (Leishmania homologue of receptor for activated C kinase) Ag has been shown to be more durable than vaccination with LACK protein plus IL-12. One mechanism to account for this may be the selective ability of DNA vaccination to induce CD8(+) IFN-gamma-producing T cells. In this regard, we previously reported that depletion of CD8(+) T cells in LACK DNA-vaccinated mice abrogated protection when infectious challenge was done 2 wk postvaccination, In this study, we extend these findings to study the mechanism by which CD8(+) T cells induced by LACK DNA vaccination mediate both short- and long-term protective immunity against L, major, Mice vaccinated with LACK DNA and depleted of CD8(+) T cells at the time of vaccination or infection were unable to control infection when challenge was done 2 or 12 wk postvaccination, Remarkably, it was noted that depletion of CD8(+) T cells in LACK DNA-vaccinated mice was associated with a striking decrease in the frequency of LACK-specific CD4(+) IFN-gamma-producing T cells both before and after infection. Moreover, data are presented to suggest a mechanism by which CD8(+) T cells exert this regulatory role. Taken together, these data provide additional insight into how Th1 cells are generated and sustained in vivo and suggest a potentially novel immunoregulatory role for CD8(+) T cells following DNA vaccination. C1 NIAID, Clin Invest Lab, Clin Immunol Sect, NIH, Bethesda, MD 20892 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. CNRS, Inst Pharmacol Mol & Cellulaire, F-06560 Valbonne, France. Univ Calif San Francisco, San Francisco, CA 94143 USA. NIH, Howard Hughes Med Inst, Bethesda, MD 20892 USA. RP Seder, RA (reprint author), NIAID, Clin Invest Lab, Clin Immunol Sect, NIH, 10 Ctr Dr,Room 10-11C215, Bethesda, MD 20892 USA. OI Prussin, Calman/0000-0002-3917-3326 NR 36 TC 103 Z9 106 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 2000 VL 165 IS 2 BP 915 EP 924 PG 10 WC Immunology SC Immunology GA 332QU UT WOS:000088085600037 PM 10878366 ER PT J AU Belkaid, Y Mendez, S Lira, R Kadambi, N Milon, G Sacks, D AF Belkaid, Y Mendez, S Lira, R Kadambi, N Milon, G Sacks, D TI A natural model of Leishmania major infection reveals a prolonged "silent" phase of parasite amplification in the skin before the onset of lesion formation and immunity SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CD4(+) T-CELLS; EXPERIMENTAL CUTANEOUS LEISHMANIASIS; NITRIC-OXIDE SYNTHASE; DENDRITIC CELLS; MEDIATED-IMMUNITY; COSTIMULATORY MOLECULES; LANGERHANS CELLS; IL-12 INDUCTION; DEFICIENT MICE; MACROPHAGES AB A model of Leishmania major infection in C57BL/6 mice has been established that combines two main features of natural transmission: low dose (100 metacyclic promastigotes) and inoculation into a dermal site (the ear dermis), The evolution of the dermal lesion could be dissociated into two distinct phases, The initial "silent" phase, lasting 4-5 wk, favored establishment of the peak load of parasites in the dermis in the absence of lesion formation or any overt histopathologic changes in the site. The second phase corresponds to the development of a lesion associated with an acute infiltration of neutrophils, macrophages, and eosinophils into the dermis and was coincident with the killing of parasites in the site. The onset of immunity/pathology was correlated with the appearance of cells staining for IL-12p40 and IFN-gamma in the epidermal compartment, and an expansion of T cells capable of producing IFN-gamma in the draining lymph node. Parasite growth was not enhanced over the first 4.5 wk in anti-CD4-treated mice, SCID mice, or C57BL/6 mice deficient in IL-12p40, IFN-gamma, CD40 ligand, or inducible NO synthase, These mice all failed to ultimately control infection in the site, but in some cases (anti-CD4 treated, IL-12p40(-/-), CD40 ligand(-/-), and SCID) high dermal parasite loads were associated with little or no pathology. These results extend to a natural infection model a role for Th1 cells in both acquired resistance and lesion formation, and document the remarkable avoidance of this response during a prolonged phase of parasite amplification in the skin . C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. Inst Pasteur, Unite Immunophysiol Cellulaire, Paris, France. RP Sacks, D (reprint author), NIAID, Parasit Dis Lab, NIH, Bldg 4,Room 126,Ctr Dr,MSC 0425, Bethesda, MD 20892 USA. NR 57 TC 231 Z9 235 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 JUL 15 PY 2000 VL 165 IS 2 BP 969 EP 977 PG 9 WC Immunology SC Immunology GA 332QU UT WOS:000088085600044 PM 10878373 ER PT J AU Gea-Banacloche, JC Migueles, SA Martino, L Shupert, WL McNeil, AC Sabbaghian, MS Ehler, L Prussin, C Stevens, R Lambert, L Altman, J Hallahan, CW de Quiros, JCLB Connors, M AF Gea-Banacloche, JC Migueles, SA Martino, L Shupert, WL McNeil, AC Sabbaghian, MS Ehler, L Prussin, C Stevens, R Lambert, L Altman, J Hallahan, CW de Quiros, JCLB Connors, M TI Maintenance of large numbers of virus-specific CD8(+) T cells in HIV-infected progressors and long-term nonprogressors SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NEUTRALIZING ANTIBODY-RESPONSES; IMMUNODEFICIENCY-VIRUS; ANTIRETROVIRAL THERAPY; LYMPHOCYTE ACTIVITY; TYPE-1 INFECTION; RECEPTOR REPERTOIRE; EFFECTOR FUNCTION; VIRAL-INFECTION; FLOW-CYTOMETRY; ELDERLY HUMANS AB The virus-specific CD8(+) T cell responses of 21 HIV-infected patients were studied including a unique cohort of long-term nonprogressors with low levels of plasma viral RNA and strong proliferative responses to HIV Ags, HIV-specific CD8(+) T cell responses were studied by a combination of standard cytotoxic T cell (CTL) assays, MHC tetramers, and TCR repertoire analysis, The frequencies of CD8(+) T cells specific to the majority of HIV gene products were measured by how cytometric detection of intracellular IFN-gamma in response to HIV-vaccinia recombinant-infected autologous B cells. Very high frequencies (0.8-18.0%) of circulating CD8(+) T cells were found to be HIV specific. High frequencies of HIV-specific CD8(+) T cells were not limited to long-tern nonprogressors with restriction of plasma virus. No correlation was found between the frequency of HIV-specific CD8(+) T cells and levels of plasma viremia, In each case, the vast majority of cells (up to 17.2%) responded to gag-pol. Repertoire analysis showed these large numbers of Ag-specific cells were scattered throughout the repertoire and in the majority of cases not contained within large monoclonal expansions. These data demonstrate that high numbers of HIV-specific CD8(+) T cells exist even in patients with high-level viremia and progressive disease. Further, they suggest that other qualitative parameters of the CD8+ T cell response may differentiate some patients with very low levels of plasma virus and nonprogressive disease. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. Sci Applicat Int Corp, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Emory Univ, Vaccine Ctr Yerkes, Atlanta, GA 30322 USA. Hosp Gen Gregorio Maranon, Microbiol Serv, Madrid, Spain. Univ Autonoma Madrid, Serv Med Interne 1, Clin Puerta de Hierro, Madrid, Spain. RP Connors, M (reprint author), NIAID, Immunoregulat Lab, NIH, Bldg 10,Room 11B-09,10 Ctr Dr,MSC 1876, Bethesda, MD 20892 USA. OI Prussin, Calman/0000-0002-3917-3326 NR 74 TC 203 Z9 206 U1 0 U2 3 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 2000 VL 165 IS 2 BP 1082 EP 1092 PG 11 WC Immunology SC Immunology GA 332QU UT WOS:000088085600058 PM 10878387 ER PT J AU Yamada, M Ariga, T Kawamura, N Yamaguchi, K Ohtsu, M Nelson, DL Kondoh, T Kobayashi, I Okano, M Kobayashi, K Sakiyama, Y AF Yamada, M Ariga, T Kawamura, N Yamaguchi, K Ohtsu, M Nelson, DL Kondoh, T Kobayashi, I Okano, M Kobayashi, K Sakiyama, Y TI Determination of carrier status for the Wiskott-Aldrich syndrome by flow cytometric analysis of Wiskott-Aldrich syndrome protein expression in peripheral blood mononuclear cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID LYMPHOCYTES; INACTIVATION; MUTATIONS; GENE AB The Wiskott-Aldrich syndrome (WAS) is caused by defects in the WAS protein (WASP) gene on the X chromosome. Previous study disclosed that flow cytometric analysis of intracellular WASP expression (FCM-WASP analysis) in lymphocytes was useful for the diagnosis of WAS patients. Lymphocytes from all WAS patients showed WASP(dim) instead of WASP(bright). Here we report that FCM-WASP analysis in monocytes could be a useful tool for the WAS carrier diagnosis. Monocytes from all nine WAS carriers showed varied population of WASP(dim) together with WASP(bright). None of control individuals possessed the WASP(dim) population. In contrast, lymphocytes from all the carriers except two lacked the WASP(dim) population. The difference of the WASP(dim) population in monocytes and lymphocytes observed in WAS carriers suggests that WASP plays a more critical role in the development of lymphocytes than in that of monocytes. The present studies suggest that a skewed X-chromosomal inactivation pattern observed in WAS carrier peripheral blood cells is not fixed at the hemopoietic stem cell level but progresses after the lineage commitment. C1 Hokkaido Univ, Sch Med, Dept Human Gene Therapy, Kita Ku, Sapporo, Hokkaido 0608638, Japan. Hokkaido Univ, Sch Med, Dept Pediat, Sapporo, Hokkaido 0608638, Japan. NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. Nagasaki Univ, Sch Med, Dept Pediat, Nagasaki 852, Japan. RP Ariga, T (reprint author), Hokkaido Univ, Sch Med, Dept Human Gene Therapy, Kita Ku, N-14 W-5, Sapporo, Hokkaido 0608638, Japan. RI Ariga, Tadashi/A-4252-2012; Kobayashi, Ichiro/G-8638-2012 NR 17 TC 26 Z9 27 U1 0 U2 3 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 2000 VL 165 IS 2 BP 1119 EP 1122 PG 4 WC Immunology SC Immunology GA 332QU UT WOS:000088085600062 PM 10878391 ER PT J AU Zeng, G Touloukian, CE Wang, X Restifo, NP Rosenberg, SA Wang, RF AF Zeng, G Touloukian, CE Wang, X Restifo, NP Rosenberg, SA Wang, RF TI Identification of CD4(+) T cell epitopes from NY-ESO-1 presented by HLA-DR molecules SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HUMAN TUMOR-ANTIGENS; IMMUNE-RESPONSE; METASTATIC MELANOMA; DENDRITIC CELLS; PEPTIDE; LYMPHOCYTES; RECOGNITION; VACCINATION; PROTEIN; MAGE-3 AB In previous studies, the shared cancer-testis Ag, NY-ESO-1, was demonstrated to be recognized by both Abs and CD8(+) T cells. Gene expression of NY-ESO-1 was detected in many tumor types, Including melanoma, breast, and lung cancers, but was not found in normal tissues, with the exception of testis, In this study, we describe the identification of MHC class II-restricted T cell epitopes from NY-ESO-1, Candidate CD4(+) T cell peptides were first identified using HLA-DR4 transgenic mice immunized with the NY-ESO-1 protein. NY-ESO-l-specific CD4(+) T cells were then generated from PBMC of a patient with melanoma stimulated with the candidate peptides in vitro. These CD4(+) T cells recognized NY-ESO-1 peptides or protein pulsed on HLA-DR4(+) EBV B cells, and also recognized tumor cells expressing HLA-DR4 and NY-ESO-I, A 10-mer peptide (VLLKEFTVSG) was recognized by CD4(+) T cells. These studies provide new opportunities for developing more effective vaccine strategies by using tumor-specific CD4(+) T cells. This approach may be applicable to the identification of CD4(+) T cell epitopes from many known tumor Ags recognized by CD8(+) T cells. C1 NIH, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Wang, RF (reprint author), Baylor Coll Med, ALKEK Bldg N1120,1 Baylor Plaza, Houston, TX 77030 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 39 TC 107 Z9 112 U1 0 U2 3 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 2000 VL 165 IS 2 BP 1153 EP 1159 PG 7 WC Immunology SC Immunology GA 332QU UT WOS:000088085600066 PM 10878395 ER PT J AU Camandola, S Mattson, MP AF Camandola, S Mattson, MP TI Pro-apoptotic action of Par-4 involves inhibition of NF-kappa B activity and suppression of Bcl-2 expression SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE apoptosis; kinase; mitochondria; neurons; neurotrophic factors ID NECROSIS-FACTOR-ALPHA; PROTEIN-KINASE-C; TROPHIC FACTOR WITHDRAWAL; AMYLOID BETA-PEPTIDE; HIPPOCAMPAL-NEURONS; ALZHEIMER-DISEASE; BINDING FACTOR; UP-REGULATION; NEURAL CELLS; PC12 CELLS AB Par-4(1) (prostate apoptosis response 4) is known to function at an early stage in apoptosis in several different cell types, including neurons. On the other hand, activation of the transcription factor NF-kappa B can prevent apoptosis in various cancer cells and neurons. We now report that overexpression of full-length Par-4 in cultured PC12 cells results in a suppression of basal NF-kappa B DNA-binding activity and NF-kappa B activation following trophic factor withdrawal (TFW). The decreased NF-kappa B activity is correlated with enhanced apoptosis. Conversely, NF-kappa B activity is increased and vulnerability to apoptosis reduced in cells overexpressing a dominant-negative form of Par-4. Par-4 overexpression or functional blockade had no effect on AP-1 DNA-binding activity. Expression of the antiapoptotic protein Bcl-2 was dramatically reduced in PC12 cells overexpressing Par-4. Our data suggest that suppression of NF-kappa B activation plays a major role in the proapoptotic function of Par-4. (C) 2000 Wiley-Liss, Inc. C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. Univ Kentucky, Sanders Brown Res Ctr Aging, Lexington, KY USA. Univ Kentucky, Dept Anat & Neurobiol, Lexington, KY USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 34 TC 66 Z9 68 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 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD JUL 15 PY 2000 VL 61 IS 2 BP 134 EP 139 DI 10.1002/1097-4547(20000715)61:2<134::AID-JNR3>3.0.CO;2-P PG 6 WC Neurosciences SC Neurosciences & Neurology GA 331AJ UT WOS:000087994500003 PM 10878585 ER PT J AU Sarafova, S Siu, G AF Sarafova, S Siu, G TI Precise arrangement of factor-binding sites is required for murine CD4 promoter function SO NUCLEIC ACIDS RESEARCH LA English DT Article ID T-CELL DEVELOPMENT; ZINC-FINGER PROTEIN; GENE-EXPRESSION; B-CELLS; TRANSCRIPTION; ENHANCER; SELECTION; MYB; MAZ; TERMINATION AB The control of CD4 expression is linked to the signaling events that mediate T-cell development and is directly dependent on the CD4 promoter. The CD4 promoter does not contain functionally redundant sites: all four factor-binding sites must be intact to achieve wild-type activity, Here we demonstrate that the precise position of three factor-binding sites relative to each other is essential for promoter activity, indicating that they function together as an inseparable cassette for assembly of the transcription initiation complex, Small changes in either phasing or distance between any two sites in this cassette leads to complete abrogation of promoter function, In addition, we demonstrate that one of the factors that bind the promoter cassette is not present in CD8 SP T-C cells, Thus, this factor is a candidate for mediating the relative subclass specificity of CD4 promoter function in activated CD4 SP T-H cells. C1 Columbia Univ, Coll Phys & Surg, Dept Microbiol, New York, NY 10032 USA. Columbia Univ, Coll Phys & Surg, Integrated Program Cellular Mol & Biophys Studies, New York, NY 10032 USA. RP Siu, G (reprint author), NIH, Bldg 10,Room 4B36,10 Ctr Dr, Bethesda, MD 20892 USA. FU NIAID NIH HHS [P30 AI42848, P30 AI042848, R01 AI034925, R01 AI34925] NR 40 TC 11 Z9 12 U1 0 U2 1 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 2000 VL 28 IS 14 BP 2664 EP 2671 DI 10.1093/nar/28.14.2664 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338PB UT WOS:000088427500006 PM 10908321 ER PT J AU Machwe, A Ganunis, R Bohr, VA Orren, DK AF Machwe, A Ganunis, R Bohr, VA Orren, DK TI Selective blockage of the 3 '-> 5 ' exonuclease activity of WRN protein by certain oxidative modifications and bulky lesions in DNA SO NUCLEIC ACIDS RESEARCH LA English DT Article ID WERNER-SYNDROME PROTEIN; SYNDROME GENE-PRODUCT; ESCHERICHIA-COLI; HELICASE ACTIVITY; RECQ FAMILY; REPLICATION; POLYMERASE; CLONING; REPAIR; SENSITIVITY AB Individuals with mutations in the WRN gene suffer from Werner syndrome, a disease with early onset of many characteristics of normal aging. The WRN protein (WRNp) functions in DNA metabolism, as the purified polypeptide has both 3'-->5' helicase and 3'-->5' exonuclease activities, In this study, we have further characterized WRNp exonuclease activity by examining its ability to degrade double-stranded DNA substrates containing abnormal and damaged nucleotides. In addition, we directly compared the 3'-->5' WRNp exonuclease activity with that of exonuclease III and the Klenow fragment of DNA polymerase I. Our results indicate that the presence of certain abnormal bases (such as uracil and hypoxanthine) does not inhibit the exonuclease activity of WRNp, exonuclease III or Klenow, whereas other DNA modifications, including apurinic sites, 8-oxoguanine, 8-oxoadenine and cholesterol adducts, inhibit or block WRNp, The ability of damaged nucleotides to inhibit exonucleolytic digestion differs significantly between WRNp, exonuclease III and Klenow, indicating that each exonuclease has a distinct mechanism of action, In addition, normal and modified DNA substrates are degraded similarly by full-length WRNp and an N-terminal fragment of WRNp, indicating that the specificity for this activity lies mostly within this region, The biochemical and physiological significance of these results is discussed. C1 NIA, Genet Mol Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. RP Bohr, VA (reprint author), NIA, Genet Mol Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 39 TC 48 Z9 48 U1 1 U2 2 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 2000 VL 28 IS 14 BP 2762 EP 2770 DI 10.1093/nar/28.14.2762 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338PB UT WOS:000088427500018 PM 10908333 ER PT J AU Schneider, TD AF Schneider, TD TI Evolution of biological information SO NUCLEIC ACIDS RESEARCH LA English DT Article ID SEQUENCE LOGOS; MOLECULAR MACHINES; CHANNEL CAPACITY; BINDING-SITES; DNA; COMPLEXITY; DISPLAY; ENERGY AB How do genetic systems gain information by evolutionary processes? Answering this question precisely requires a robust, quantitative measure of information. Fortunately, 50 years ago Claude Shannon defined information as a decrease in the uncertainty of a receiver. For molecular systems, uncertainty is closely related to entropy and hence has clear connections to the Second Law of Thermodynamics. These aspects of information theory have allowed the development of a straightforward and practical method of measuring information in genetic control systems. Here this method is used to observe information gain in the binding sites for an artificial 'protein' in a computer simulation of evolution. The simulation begins with zero information and, as in naturally occurring genetic systems, the information measured in the fully evolved binding sites is close to that needed to locate the sites in the genome. The transition is rapid, demonstrating that information gain can occur by punctuated equilibrium. C1 NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, Frederick, MD 21702 USA. RP Schneider, TD (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, POB B, Frederick, MD 21702 USA. OI Schneider, Thomas/0000-0002-9841-1531 NR 37 TC 73 Z9 74 U1 0 U2 14 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 2000 VL 28 IS 14 BP 2794 EP 2799 DI 10.1093/nar/28.14.2794 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338PB UT WOS:000088427500022 PM 10908337 ER PT J AU Grabczyk, E Usdin, K AF Grabczyk, E Usdin, K TI The GAA center dot TTC triplet repeat expanded in Friedreich's ataxia impedes transcription elongation by T7 RNA polymerase in a length and supercoil dependent manner SO NUCLEIC ACIDS RESEARCH LA English DT Article ID HOMOPURINE-HOMOPYRIMIDINE SEQUENCES; DNA STRUCTURES; PLASMID DNA; H-DNA; EXPANSION; TEMPLATE; TERMINATION; EVOLUTION; MECHANISM; FRATAXIN AB Large expansions of the trinucleotide repeat GAA . TTC within the first intron of the X25 (frataxin) gene cause Friedreich's ataxia, the most common inherited ataxia, Expansion leads to reduced levels of frataxin mRNA in affected individuals. Here we show that GAA . TTC tracts, in the absence of any other frataxin gene sequences, can reduce the amount of GAA-containing transcript produced in a defined in vitro transcription system. This effect is due to an impediment to elongation that forms in the GAA . TTC tract during transcription, a phenomenon that is exacerbated by both superhelical stress and increased tract length. On supercoiled templates the major truncations of the GAA-containing transcripts occur in the distal (3') end of the GAA repeat, To account for these observations we present a model in which an RNA polymerase advancing within a long GAA . TTC tract initiates the transient formation of an R . R . Y intramolecular DNA tripler. The non-template (GAA) strand folds back creating a loop in the template strand, and the distal triplex-duplex junction. C1 NIDDK, Sect Genom Struct & Funct, Mol & Cellular Biol Lab, NIH, Bethesda, MD 20892 USA. RP Grabczyk, E (reprint author), NIDDK, Sect Genom Struct & Funct, Mol & Cellular Biol Lab, NIH, Bldg 8,Room 202,8 Ctr DR MSC 0830, Bethesda, MD 20892 USA. NR 40 TC 143 Z9 147 U1 2 U2 8 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 2000 VL 28 IS 14 BP 2815 EP 2822 DI 10.1093/nar/28.14.2815 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338PB UT WOS:000088427500025 PM 10908340 ER PT J AU Drew, LR Tang, DC Berg, PE Rodgers, GP AF Drew, LR Tang, DC Berg, PE Rodgers, GP TI The role of trans-acting factors and DNA-bending in the silencing of human beta-globin gene expression SO NUCLEIC ACIDS RESEARCH LA English DT Article ID LOCUS-CONTROL REGION; KRUPPEL-LIKE FACTOR; LEUKEMIC CELL-LINE; REGULATORY PROTEINS; ERYTHROID-CELLS; BINDING DOMAIN; HMG PROTEINS; TRANSCRIPTION; BOX; COMPLEX AB The molecular mechanisms which govern the developmental specificity of human beta-globin gene transcription have been studied in K562 cells, a human eyrthro-leukemia line that expresses minimal beta-globin, Protein-binding analysis reveals that the 5' region contains three elements bound by trans-acting factors, beta-protein 1 (BP1) and beta-protein 2 (BP2), In vitro mutagenesis of each individual element in a beta-globin vector containing chloramphenicol acetyltransferase (pCAT) followed by transient transfection into K562 cells increased levels of CAT activity 5.5-fold higher than wild-type (wt) beta CAT, consistent with their silencing role. Mutagenesis of all three elements, however, resulted in activity significantly lower than wt beta CAT, BP1 and BP2 motifs have overlapping binding sites for high mobility group proteins (HMG1+2), DNA-bending factors, shown here to extrinsically bend the beta-globin promoter. Theoretically, mutations in all beta-protein binding sites could affect the binding of HMG1+2 sufficiently to impede DNA-protein and/or protein-protein interactions needed to facilitate constitutive gene expression. Placing two turns of DNA between BP1 and BP2 motifs also increased expression 3-fold, indicative of spatial constraints required for optimal silencing. However, insertion of the HMG1+2 DNA-bending motif (also equivalent to two turns) facilitates beta-silencing by re-establishment of BP1-BP2 proximity. Thus a combination of general DNA-bending and specific transcriptional factors appear to be involved in beta-globin silencing in the embryonic/fetal erythroid stage. C1 NIDDKD, Mol & Clin Hematol Branch, NIH, Bethesda, MD 20892 USA. George Washington Univ, Med Ctr, Dept Biochem & Mol Biol, Washington, DC 20037 USA. RP Rodgers, GP (reprint author), NIDDKD, Mol & Clin Hematol Branch, NIH, Bldg 10,Room 9N115,10 Ctr Dr, Bethesda, MD 20892 USA. FU NIDDK NIH HHS [DK53533] NR 45 TC 8 Z9 8 U1 0 U2 0 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 2000 VL 28 IS 14 BP 2823 EP 2830 DI 10.1093/nar/28.14.2823 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 338PB UT WOS:000088427500026 PM 10908341 ER PT J AU Boguna, M Berezhkovskii, AM Weiss, GH AF Boguna, M Berezhkovskii, AM Weiss, GH TI Residence time densities for non-Markovian systems. (I). The two-state system SO PHYSICA A LA English DT Article ID MOLECULE FLUORESCENCE SPECTROSCOPY; SINGLE MOLECULES; CONFORMATIONAL FLUCTUATIONS; TEMPERATURE; PENETRATION; TISSUE AB We study dynamical system which makes transitions between two states at random times. We analyze properties of the cumulative time tau spent by the system in a given state up to time T. When the probability density for the residence time in a single sojourn in the given state differs from a negative exponential the system will be non-Markovian. Simple analytical expressions are derived for the Laplace transform with respect to T of moments of the cumulative residence time. An exact Fourier-Laplace transform of the probability densities for tau at a fixed T are also found. It can be inferred from this expression, that at sufficiently large T the probability densities tend towards a Gaussian. The parameters that define the Gaussian are also given. (C) 2000 Published by Elsevier Science B.V. All rights reserved. C1 NIH, Math & Stat Comp Lab, Div Comp Res & Technol, Bethesda, MD 20892 USA. NICHHD, NIH, Bethesda, MD 20892 USA. Karpov Inst Phys Chem, Moscow 103064, Russia. RP Weiss, GH (reprint author), NIH, Math & Stat Comp Lab, Div Comp Res & Technol, Bldg 12 A Room 2007, Bethesda, MD 20892 USA. RI Boguna, Marian/B-7795-2011 OI Boguna, Marian/0000-0001-7833-3487 NR 37 TC 28 Z9 28 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4371 J9 PHYSICA A JI Physica A PD JUL 15 PY 2000 VL 282 IS 3-4 BP 475 EP 485 DI 10.1016/S0378-4371(00)00091-1 PG 11 WC Physics, Multidisciplinary SC Physics GA 330GZ UT WOS:000087955600009 ER PT J AU Boguna, M Weiss, GH AF Boguna, M Weiss, GH TI Residence time densities for non-Markovian systems. (II). The N-state system SO PHYSICA A LA English DT Article ID MOLECULE FLUORESCENCE SPECTROSCOPY; SINGLE AB In the preceding paper (Boguna et al., Physica A 282 (2000) 475), we developed formalism to calculate a probability density for the cumulative time spent in one out of a two-state non-Markovian system of the form S-1 reversible arrow S-2 when the system is observed continuously for a time T. The asymptotic form for the probability density was shown to be Gaussian. In this paper we indicate how to reduce any reacting nearest-neighbor system, i.e., S-1 reversible arrow S-2 reversible arrow...reversible arrow S-N to an equivalent two-state system. This also shows that the probability density for the cumulative residence time in a single state is asymptotic to a Gaussian. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIH, Math & Stat Comp Lab, Div Comp Res & Technol, Bethesda, MD 20892 USA. RP Weiss, GH (reprint author), NIH, Math & Stat Comp Lab, Div Comp Res & Technol, Bldg 12 A Room 2007, Bethesda, MD 20892 USA. RI Boguna, Marian/B-7795-2011 OI Boguna, Marian/0000-0001-7833-3487 NR 17 TC 11 Z9 11 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4371 J9 PHYSICA A JI Physica A PD JUL 15 PY 2000 VL 282 IS 3-4 BP 486 EP 494 DI 10.1016/S0378-4371(00)00092-3 PG 9 WC Physics, Multidisciplinary SC Physics GA 330GZ UT WOS:000087955600010 ER PT J AU Engels, EA Schmid, CH Terrin, N Olkin, I Lau, J AF Engels, EA Schmid, CH Terrin, N Olkin, I Lau, J TI Heterogeneity and statistical significance in meta-analysis: an empirical study of 125 meta-analyses SO STATISTICS IN MEDICINE LA English DT Article ID ACUTE MYOCARDIAL-INFARCTION; RANDOMIZED CLINICAL-TRIALS; METAANALYSIS; MAGNESIUM; EFFICACY; NUMBER; CANCER AB For meta-analysis, substantial uncertainty remains about the most appropriate statistical methods for combining the results of separate trials. An important issue for meta-analysis is how to incorporate heterogeneity, defined as variation among the results of individual trials beyond that expected from chance, into summary estimates of treatment effect. Another consideration is which 'metric' to use to measure treatment effect; for trials with binary outcomes, there are several possible metrics, including the odds ratio (a relative measure) and risk difference tan absolute measure). To examine empirically how assessment of treatment effect and heterogeneity may differ when different methods are utilized, we studied 125 meta-analyses representative of those performed by clinical investigators. There was no meta-analysis in which the summary risk difference and odds ratio were discrepant to the extent that one indicated significant benefit while the other indicated significant harm. Further, for most meta-analyses, summary odds ratios and risk differences agreed in statistical significance, leading to similar conclusions about whether treatments affected outcome. Heterogeneity was common regardless of whether treatment effects were measured by odds ratios or risk differences. However, risk differences usually displayed more heterogeneity than odds ratios. Random effects estimates, which incorporate heterogeneity, tended to be less precisely estimated than fixed effects estimates. We present two exceptions to these observations, which derive from the weights assigned to individual trial estimates. We discuss the implications of these findings for selection of a metric for meta-analysis and incorporation of heterogeneity into summary estimates. Published in 2000 by John Wiley & Sons, Ltd. C1 Tufts Univ, Sch Med, New England Med Ctr, Dept Med,Div Clin Care Res, Boston, MA 02111 USA. Tufts Univ, Sch Med, New England Med Ctr, Biostat Res Ctr, Boston, MA 02111 USA. Stanford Univ, Dept Stat, Stanford, CA 94305 USA. RP Engels, EA (reprint author), NCI, Viral Epidemiol Branch, Div Canc Epidemiol & Genet, 6120 Execut Blvd,MSC 7248, Rockville, MD 20852 USA. OI Schmid, Christopher/0000-0002-0855-5313 FU AHRQ HHS [HS10064, R01HS08532, T32HS00060] NR 40 TC 188 Z9 190 U1 0 U2 16 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0277-6715 J9 STAT MED JI Stat. Med. PD JUL 15 PY 2000 VL 19 IS 13 BP 1707 EP 1728 DI 10.1002/1097-0258(20000715)19:13<1707::AID-SIM491>3.0.CO;2-P PG 22 WC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Medicine, Research & Experimental; Statistics & Probability SC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Research & Experimental Medicine; Mathematics GA 332FF UT WOS:000088063000001 PM 10861773 ER PT J AU Cronin, KA Feuer, EJ AF Cronin, KA Feuer, EJ TI Cumulative cause-specific mortality for cancer patients in the presence of other causes: a crude analogue of relative survival SO STATISTICS IN MEDICINE LA English DT Article ID FAILURE PROBABILITIES; COMPETING RISKS; IMPACT AB A common population-based cancer progress measure for net survival (survival in the absence of other causes) of cancer patients is relative survival. Relative survival is defined as the ratio of a population of observed survivors in a cohort of cancer patients to the proportion of expected survivors in a comparable set of cancer-free individuals in the general public, thus giving a measure of excess mortality due to cancer. Relative survival was originally designed to address the question of whether or not there is evidence that patients have been cured. It has proven to be a useful survival measure in several areas, including the evaluation of cancer control efforts and the application of cure models. However, it is not representative of the actual survival patterns observed in a cohort of cancer patients. This paper suggests a measure for cumulative crude tin the presence of other causes) cause-specific probability of death for a population diagnosed with cancer. The measure does not use cause of death information which can be unreliable for population cancer registries. Point estimates and variances are derived for crude cause-specific probability of death using relative survival instead of cause of death information. Examples are given for men diagnosed with localized prostate cancer over the age of 70 and women diagnosed with regional breast cancer using Surveillance, Epidemiology and End Results (SEER) Program data. The examples emphasize the differences in crude and net mortality measures and suggest areas where a crude measure is more informative. Estimates of this type are especially important for older patients as new screening modalities detect cancers earlier and choice of treatment or even 'watchful waiting' become viable options. Published in 2000 by John Wiley & Sons, Ltd. C1 NCI, Appl Res Branch, Bethesda, MD 20892 USA. RP Cronin, KA (reprint author), NCI, Appl Res Branch, EPN 4103,6130 Execut Blvd, Bethesda, MD 20892 USA. EM cronink@dcpcepn.nci.nih.gov NR 21 TC 55 Z9 56 U1 0 U2 5 PU WILEY-BLACKWELL PI MALDEN PA COMMERCE PLACE, 350 MAIN ST, MALDEN 02148, MA USA SN 0277-6715 J9 STAT MED JI Stat. Med. PD JUL 15 PY 2000 VL 19 IS 13 BP 1729 EP 1740 DI 10.1002/1097-0258(20000715)19:13<1729::AID-SIM484>3.0.CO;2-9 PG 12 WC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Medicine, Research & Experimental; Statistics & Probability SC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Research & Experimental Medicine; Mathematics GA 332FF UT WOS:000088063000002 PM 10861774 ER PT J AU Horne, MK Goad, JL Merryman, PK Cullinane, AM AF Horne, MK Goad, JL Merryman, PK Cullinane, AM TI Comparison of the effect of histidine-rich glycoprotein and 6-aminohexanoic acid on plasmin production and fibrinolysis in vitro SO THROMBOSIS RESEARCH LA English DT Article DE histidine-rich glycoprotein; fibrinolysis; 6-aminohexanoic acid ID ACTIVATION; MECHANISM; AFFINITY; KINETICS; BINDING AB Because histidine-rich glycoprotein binds to the kringle 1-3 domain of plasminogen. it may affect fibrinolysis by reducing fibrin-dependent plasmin production, and in this way it could be mechanistically analogous to 6-aminohexanoic acid. We tested this hypothesis by comparing the effects of histidine-rich glycoprotein and 6-aminohexanoic acid in an in vitro assay of fibrin-dependent plasmin production mediated by tissue plasminogen activator. Whereas 1 mM of 6-aminohexanoic acid increased the K-m of the reaction from similar to 0.22 mu M to similar to 1.7 mu M, 2 mu M of histidine-rich glycoprotein had no discernible effect. Similar results were obtained in an assay based upon fibrin clot lysis. Therefore, we could not document an effect of histidine-rich glycoprotein on the rate of fibrin-dependent plasmin production. Published by Elsevier Science Ltd. C1 NIH, Hematol Serv, Dept Clin Pathol, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Horne, MK (reprint author), NIH, Hematol Serv, Dept Clin Pathol, Warren G Magnuson Clin Ctr, Room 2C306,Bldg 10, Bethesda, MD 20892 USA. NR 25 TC 6 Z9 6 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0049-3848 J9 THROMB RES JI Thromb. Res. PD JUL 15 PY 2000 VL 99 IS 2 BP 179 EP 186 DI 10.1016/S0049-3848(00)00231-0 PG 8 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 347EU UT WOS:000088915200009 PM 10946092 ER PT J AU Men, R Wyatt, L Tokimatsu, I Arakaki, S Shameem, G Elkins, R Chanock, R Moss, B Lai, CJ AF Men, R Wyatt, L Tokimatsu, I Arakaki, S Shameem, G Elkins, R Chanock, R Moss, B Lai, CJ TI Immunization of rhesus monkeys with a recombinant of modified vaccinia virus Ankara expressing a truncated envelope glycoprotein of dengue type 2 virus induced resistance to dengue type 2 virus challenge SO VACCINE LA English DT Article DE modified vaccinia virus Ankara; dengue virus; envelope protein; protective immunity ID JAPANESE ENCEPHALITIS-VIRUS; TICK-BORNE ENCEPHALITIS; STRUCTURAL PROTEINS; NONSTRUCTURAL PROTEIN-NS1; INCREASED IMMUNOGENICITY; MICE; VOLUNTEERS; EFFICACY; STRAIN; GENES AB Dengue epidemics increasingly pose a public health problem in most countries of the tropical and subtropical areas. Despite decades of research, development of a safe and effective live dengue virus vaccine is still at the experimental stage. To explore an alternative vaccine strategy, we employed the highly attenuated, replication-deficient modified vaccinia Ankara (MVA) as a vector to construct recombinants for expression of the major envelope glycoprotein of one or more dengue virus serotypes. MVA recombinants expressing the highly immunogenic C-terminally truncated dengue type 2 virus (DEN2) or dengue type 4 virus (DEN4) envelope protein (E), approx. 80% of the full-length, were evaluated for their protective immunity in animal models. Each of these recombinants elicited an elevated antibody response to DEN2 or DEN4 E in mice following the booster inoculation, as detected by radio-immunoprecipitation. Recombinant MVA-DEN2 80%E, but not MVA-DEN4 80%E, induced a neutralizing antibody response. The MVA-DEN2 80%E recombinant was chosen to further evaluate its ability to induce resistance to wild type DEN2 challenge in monkeys. Monkeys immunized twice with recombinant MVA-DEN2 80%E developed a low to moderate antibody response and were partially protected against DEN2 challenge, as determined by the viremia pattern. Importantly, the subsequent study showed that all four monkeys immunized with the recombinant in a three dose schedule developed an increased level of antibodies and were completely protected against DEN2 challenge. The potential efficacy of recombinant MVA-DEN2 80%E to protect primates against dengue infection suggests that construction and evaluation of MVA recombinants expressing other serotypes of dengue virus E for use in a tetravalent vaccine strategy might be warranted. Published by Elsevier Science Ltd. C1 NIAID, Infect Dis Lab, Bethesda, MD 20892 USA. NIAID, Viral Dis Lab, Bethesda, MD 20892 USA. RP Lai, CJ (reprint author), NIAID, Infect Dis Lab, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 31 TC 72 Z9 77 U1 0 U2 2 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0264-410X J9 VACCINE JI Vaccine PD JUL 15 PY 2000 VL 18 IS 27 BP 3113 EP 3122 DI 10.1016/S0264-410X(00)00121-3 PG 10 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 336UP UT WOS:000088321600006 PM 10856791 ER PT J AU Nagababu, E Rifkind, JM AF Nagababu, E Rifkind, JM TI Heme degradation during autoxidation of oxyhemoglobin SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE hemoglobin; autoxidation; heme degradation; fluorescent products; hydrogen peroxide; ferrylhemoglobin ID RED-BLOOD-CELLS; HYDROGEN-PEROXIDE; UNSTABLE HEMOGLOBINS; LIPID-PEROXIDATION; ERYTHROCYTE; SUPEROXIDE; OXYGEN; IRON; SUBSTITUTES; OXIDATION AB Two fluorescent heme degradation compounds are detected during autoxidation of oxyhemoglobin. These fluorescent compounds are similar to fluorescent compounds formed when hydrogen peroxide reacts with hemoglobin EE. Nagababu and J, M. Rifkind, Biochem. Biophys. Res. Commun. 247, 592-596 (1998)]. Low levels of heme degradation in the presence of superoxide and catalase are attributed to a reaction involving the superoxide produced during autoxidation. The inhibition of most of the degradation by catalase suggests that the hydrogen peroxide generated during autoxidation of oxyhemoglobin produces heme degradation by the same mechanism as the direct addition of hydrogen peroxide to hemoglobin, The formation of the fluorescent degradation products was inhibited by the peroxidase substrate, ABTS, which reduces ferrylhemoglobin to methemoglobin, indicating that ferrylhemoglobin is produced during the autoxidation of hemoglobin. It is the transient formation of this highly reactive Fe(IV) hemoglobin, which is responsible for most of the heme degradation during autoxidation. (C) 2000 Academic Press. C1 NIA, Cellular & Mol Biol Lab, NIH, Baltimore, MD 21224 USA. RP Rifkind, JM (reprint author), NIA, Cellular & Mol Biol Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 30 TC 57 Z9 57 U1 0 U2 4 PU ACADEMIC PRESS INC 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 14 PY 2000 VL 273 IS 3 BP 839 EP 845 DI 10.1006/bbrc.2000.3025 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 337LX UT WOS:000088363700009 PM 10891334 ER PT J AU Shah, PS Bizik, F Dukor, RK Qasba, PK AF Shah, PS Bizik, F Dukor, RK Qasba, PK TI Active site studies of bovine alpha 1 -> 3-galactosyltransferase and its secondary structure prediction SO BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY LA English DT Article DE glycosyltransferase; alpha 1 -> 3-galactosyltransferase; Fourier transform infrared spectroscopy ID TRANSFORM INFRARED-SPECTROSCOPY; VIBRATIONAL CIRCULAR-DICHROISM; BLOOD-GROUP-A; SUBSTRATE-SPECIFICITY; GALACTOSYL EPITOPES; AQUEOUS-SOLUTIONS; ESCHERICHIA-COLI; GROUP-B; GALACTOSYLTRANSFERASE; CDNA AB The catalytic domain of bovine alpha 1 --> 3-galactosyltransferase (alpha 3GalT), residues 80-368, have been cloned and expressed, in Escherichia coli. Using a sequential purification protocol involving a Ni2+ affinity column followed by a UDP-hexanolamine affinity column, we have obtained a pure and active protein from the soluble fraction which catalyzes the transfer of galactose (Gal) from UDP-Gal to N-acetyllactosamine (LacNAc) with a specific activity of 0.69 pmol/min/ng. The secondary structural content of alpha 3GalT protein was analyzed by Fourier transform infrared (FTIR) spectroscopy, which shows that the enzyme has about 35% beta-sheet and 22% alpha-helix. This predicted secondary structure content by FTIR spectroscopy was used in the protein sequence analysis algorithm, developed by the Biomolecular Engineering Research Center at Boston University and Tasc Inc., for the assignment of secondary structural elements to the amino acid sequence of alpha 3GalT. The enzyme appears to have three major and three minor helices and five sheet-like structures. The studies on the acceptor substrate specificity of the enzyme, alpha 3GalT, show that in addition to LacNAc, which is the natural substrate, the enzyme accepts various other disaccharides as substrates such as lactose and Gal derivatives, beta-O-methylgalactose and beta-D-thiogalactopyranoside, albeit with lower specific activities. There is an absolute requirement for Gal to be at the non-reducing end of the acceptor molecule which has to be beta 1 --> 4-linked to a second residue that can be more diverse in structure. The kinetic parameters for four acceptor molecules were determined. Lactose binds and functions in a similar way as LacNAc. However. beta-O-methylgalactose and Gal do not bind as tightly as LacNAc or lactose, as their K-ia and K-A values indicate, suggesting that the second monosaccharide is critical for holding the acceptor molecule in place. The 2' and 4' hydroxyl groups of the receiving Gal moiety are important in binding. Even though there is large structural variability associated with the second residue of the acceptor molecule, there are constraints which do not allow certain Gal-R sugars to be good acceptors for the enzyme. The beta 1 --> 4-linked residue at the second position of the acceptor molecule is preferred, but the interactions between the enzyme and the second residue are likely to be non-specific. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Struct Glycobiol Sect, Lab Expt & Computat Biol, FCRDC, Frederick, MD 21702 USA. Vysis Inc, Downers Grove, IL USA. RP Qasba, PK (reprint author), NCI, Struct Glycobiol Sect, Lab Expt & Computat Biol, FCRDC, Room 221,Bldg 469, Frederick, MD 21702 USA. NR 53 TC 8 Z9 8 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4838 J9 BBA-PROTEIN STRUCT M JI Biochim. Biophys. Acta-Protein Struct. Molec. Enzym. PD JUL 14 PY 2000 VL 1480 IS 1-2 BP 222 EP 234 DI 10.1016/S0167-4838(00)00074-1 PG 13 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 339CA UT WOS:000088459000022 PM 11004566 ER PT J AU Nishimura, Y Ito, T Hoe, KL Saavedra, JM AF Nishimura, Y Ito, T Hoe, KL Saavedra, JM TI Chronic peripheral administration of the angiotensin II AT(1) receptor antagonist Candesartan blocks brain AT(1) receptors SO BRAIN RESEARCH LA English DT Article DE renin angiotensin system; angiotensin II receptor subtypes; angiotensin II; receptor antagonists; autoradiography; in situ hybridization ID SPONTANEOUSLY HYPERTENSIVE RATS; IN-VITRO AUTORADIOGRAPHY; TYPE-1 RECEPTOR; BINDING-SITES; LOSARTAN; BARRIER; CROSS; IRBESARTAN; EXPRESSION; INHIBITION AB Brain Angiotensin II, through stimulation of brain AT(1) receptors, regulates pituitary hormones and autonomic activity. We have administered the insurmountable AT(1) antagonist Candesartan, s.c. via osmotic minipumps for 14 days, to determine whether peripheral chronic AT(1) blockade affects AT(1) receptor binding and mRNA in the brain. Peripherally administered Candesartan (0.1, 0.5 or 1.0 mg/kg per day) inhibits AT(1) binding in adrenal gland zona glomerulosa and kidney glomeruli. In addition, Candesartan dose-dependently decreases AT(1) binding in brain areas outside (subfornical organ and area postrema) and inside (paraventricular nucleus of the hypothalamus and nucleus of the solitary tract) the blood-brain barrier. Conversely, peripheral treatment with Candesattan does not affect AT(1A) receptor mRNA, the predominant receptor subtype expressed in these areas, or Angiotensin II binding to AT(1) receptors in the locus coeruleus or inferior olive,Our results demonstrate that chronic peripheral treatment with selective, potent AT(1) antagonists not only inhibits peripheral but also brain AT(1) receptors. These central effects may play a role in the antihypertensive effects of the AT(1) antagonist Candesartan. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIMH, IRP, Pharmacol Sect, Bethesda, MD 20892 USA. RP Saavedra, JM (reprint author), NIMH, IRP, Pharmacol Sect, 10 Ctr Dr,Bldg 10,Room 2D-57, Bethesda, MD 20892 USA. NR 34 TC 102 Z9 104 U1 0 U2 0 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 14 PY 2000 VL 871 IS 1 BP 29 EP 38 DI 10.1016/S0006-8993(00)02377-5 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 336BF UT WOS:000088281500004 PM 10882779 ER PT J AU Endo, K Takeshita, T Kasai, H Sasaki, Y Tanaka, N Asao, H Kikuchi, K Yamada, M Chen, M O'Shea, JJ Sugamura, K AF Endo, K Takeshita, T Kasai, H Sasaki, Y Tanaka, N Asao, H Kikuchi, K Yamada, M Chen, M O'Shea, JJ Sugamura, K TI STAM2, a new member of the STAM family, binding to the Janus kinases SO FEBS LETTERS LA English DT Article DE IL-2; GM-CSF; signal transduction; tyrosine kinase ID TRANSDUCING ADAPTER MOLECULE; COLONY-STIMULATING FACTOR; COMMON GAMMA-CHAIN; SIGNAL-TRANSDUCTION; C-MYC; POSSIBLE INVOLVEMENT; CYTOKINE RECEPTORS; CELL-GROWTH; PROTEIN; ACTIVATION AB We here cloned a cDNA encoding STAM2, a new member of the STAM family, which contains an SH3 domain and ITAM, STAM2 like STAM1 is associated with Jak2 and Jak3, and involved in the signaling for DNA synthesis and c-myc induction mediated by IL-2 and GM-CSF. Go-expression of the SH3 deletion mutants of STAM1 and STAM2 induces an additive effect on suppressing DNA synthesis upon stimulation with IL-2 and GM-CSF, suggesting that STAM1 and STAM2 exhibit compensatory effects on the signaling pathways downstream of Jak2 and Jak3 upon stimulation with GM-SCF and IL-2, respectively, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 Tohoku Univ, Sch Med, Dept Microbiol & Immunol, Aoba Ku, Sendai, Miyagi 9808575, Japan. NIAMSD, Lymphocyte Cell Biol Sect, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. Japan Sci & Technol Corp, CREST, Tokyo, Japan. RP Sugamura, K (reprint author), Tohoku Univ, Sch Med, Dept Microbiol & Immunol, Aoba Ku, Seiryo Machi 2-1, Sendai, Miyagi 9808575, Japan. RI YAMADA, MITSUHIRO/Q-8003-2016; Kikuchi, Kazu/A-4097-2017 OI YAMADA, MITSUHIRO/0000-0002-0832-1939; Kikuchi, Kazu/0000-0002-4681-4275 NR 24 TC 54 Z9 60 U1 2 U2 4 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 14 PY 2000 VL 477 IS 1-2 BP 55 EP 61 DI 10.1016/S0014-5793(00)01760-9 PG 7 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 337AM UT WOS:000088335200010 PM 10899310 ER PT J AU Zhou, XL Espey, MG Chen, JX Hofseth, LJ Miranda, KM Hussain, SP Wink, DA Harris, CC AF Zhou, XL Espey, MG Chen, JX Hofseth, LJ Miranda, KM Hussain, SP Wink, DA Harris, CC TI Inhibitory effects of nitric oxide and nitrosative stress on dopamine-beta-hydroxylase SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NG-MONOMETHYL ARGININE; ANOCOCCYGEUS MUSCLE; STRIATAL SLICES; H-3 DOPAMINE; RELEASE; RAT; RELAXATION; KINETICS; INSIGHTS; ASSAY AB Dopamine-beta-hydroxylase (D beta H) is a copper-containing enzyme that uses molecular oxygen and ascorbate to catalyze the addition of a hydroxyl group on the beta-carbon of dopamine to form norepinephrine. While norepinephrine causes vasoconstriction following reflex sympathetic stimulation, nitric oxide (NO) formation results in vasodilatation via a guanylyl cyclase-dependent mechanism. In this report, we investigated the relationship between NO and D beta H enzymatic activity. In the initial in vitro experiments, the activity of purified D beta H was inhibited by the NO donor, diethylamine/NO (DEA/NO), with an IC50 of 1 mM. The inclusion of either azide or GSH partially restored D beta H activity, suggesting the involvement of the reactive nitrogen oxide species, N2O3. Treatment of human neuroblastoma cells (SK-N-MC) with diethylamine/NO decreased cellular D beta H activity without affecting their growth rate and was augmented by the depletion of intracellular GSH. Coculture of the SK-N-MC cells with interferon-gamma and lipopolysaccharide-activated macrophages, which release NO, also reduced the D beta H activity in the neuroblastoma cells. Our results are consistent with the hypothesis that nitrosative stress, mediated by N2O3, can result in the inhibition of norepinephrine biosynthesis and may contribute to the regulation of neurotransmission and vasodilatation. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. NCI, Radiat Biol Branch, NIH, Bethesda, MD 20892 USA. RP Harris, CC (reprint author), NCI, Human Carcinogenesis Lab, NIH, Bldg 37,Rm 2C05,37 Convent Dr, Bethesda, MD 20892 USA. RI Miranda, Katrina/B-7823-2009 NR 36 TC 26 Z9 26 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 14 PY 2000 VL 275 IS 28 BP 21241 EP 21246 DI 10.1074/jbc.M904498199 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 335EM UT WOS:000088230600039 PM 10887204 ER PT J AU Vitale, N Pacheco-Rodriguez, G Ferrans, VJ Riemenschneider, W Moss, J Vaughan, M AF Vitale, N Pacheco-Rodriguez, G Ferrans, VJ Riemenschneider, W Moss, J Vaughan, M TI Specific functional interaction of human cytohesin-1 and ADP-ribosylation factor domain protein (ARD1) SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GUANINE-NUCLEOTIDE-EXCHANGE; GTPASE-ACTIVATING DOMAIN; SEC7 DOMAIN; FACTOR ARNO; CHOLERA-TOXIN; ARF GTPASE; BINDING; GOLGI; GDP; TRANSPORT AB Activation of ADP-ribosylation factors (ARFs) is mediated by guanine nucleotide-exchange proteins, which accelerate conversion of inactive ARF-GDP to active ARF-GTP. ARF domain protein (ARD1), a 64-kDa GTPase with a C-terminal ADP-ribosylation factor domain, is localized to lysosomes and the Golgi apparatus. When ARD1 was used as bait to screen a human liver cDNA library using the yeast two-hybrid system, a cDNA for cytohesin-1, a similar to 50-kDa protein with ARF guanine nucleotide-exchange protein activity, was isolated. In this system, ARD1-GDP interacted well with cytohesin-1 but very poorly with cytohesin-2. In agreement, cytohesin-1, but not cytohesin-2, markedly accelerated [S-35]guanosine 5'-3-O-(thio)triphosphate binding to ARD1. The effector region of the ARF domain of ARD1 appeared to be critical for the specific interaction with cytohesin-1. Replacement of single amino acids in the Sec7 domains of cytohesin-1 and -2 showed that residue 30 is critical for specificity. In transfected COS-7 cells, overexpressed ARD1 and cytohesin-1 were partially colocalized, as determined by confocal fluorescence microscopy. It was concluded that cytohesin-1 is likely to be involved in ARD1 activation, consistent with a role for ARD1 in the regulation of vesicular trafficking. C1 NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Pacheco-Rodriguez, G (reprint author), NHLBI, Pulm Crit Care Med Branch, NIH, Rm 5N-307,Bldg 10,10 Ctr Dr,MSC 1434, Bethesda, MD 20892 USA. RI Vitale, nicolas/G-5967-2014 OI Vitale, nicolas/0000-0002-4752-4907 NR 48 TC 10 Z9 12 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 14 PY 2000 VL 275 IS 28 BP 21331 EP 21339 DI 10.1074/jbc.M909642199 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 335EM UT WOS:000088230600051 PM 10748148 ER PT J AU Jing, NJ Marchand, C Liu, J Mitra, R Hogan, ME Pommier, Y AF Jing, NJ Marchand, C Liu, J Mitra, R Hogan, ME Pommier, Y TI Mechanism of inhibition of HIV-1 integrase by G-tetrad-forming oligonucleotides in vitro SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; CARBOXYL-TERMINAL DOMAINS; DNA-BINDING DOMAIN; CATALYTIC DOMAIN; ACTIVE-SITE; I INTEGRASE; PROTEIN; CORE; MULTIMERIZATION; IDENTIFICATION AB The G-tetrad-forming oligonucleotides T30177 and T30695 have been identified as potent inhibitors of human immunodeficiency virus type 1 integrase (HIV-1 IN) activity (Rando, R. F., Ojwang, J., Elbaggari, A., Reyes, G. R., Tinder, R., McGrath, M. S., and Hogan, M. E. (1995) J. Biol. Chem. 270, 1754-1760; Mazumder, A., Neamati, N., Ojwang, J. O., Sunder, S., Rando, R. F., and Pommier, Y. (1996) Biochemistry 35, 13762-13771; Jing, N., and Hogan, M. E. (1998) J. Biol. Chem. 273, 34992-34999). To understand the inhibition of HIV-1 IN activity by the G-quartet inhibitors, we have designed the oligonucleotides T40215 and T40216, composed of three and four G-quartets with stem lengths of 19 and 24 Angstrom, respectively. The fact that increasing the G-quartet stem length from 15 to 24 Angstrom kept inhibition of HIV-1 IN activity unchanged suggests that the binding interaction occurs between a GTGT loop domain of the G-quartet inhibitors and a catalytic site of HIV-1 ZN, referred to as a face-to-face interaction. Docking the NMR structure of T30695 (Jing and Hogan (1998)) into the x-ray structure of the core domain of HIV-1 IN, HIV-1 IN-(51-209) (Maignan, S., Guilloteau, J.-P., Qing, Z.-L., Clement-Mella, C., and Mikol, V. (1998) J. Mel. Biol. 282, 359-368), was performed using the GRAMM program. The statistical distributions of hydrogen bonding between HIV-1 IN and T30695 were obtained from the analyses of 1000 random docking structures. The docking results show a high probability of interaction between the GTGT loop residues of the G-quartet inhibitors and the catalytic site of HIV-1 IN, in agreement with the experimental observation. C1 Baylor Coll Med, Dept Mol Physiol & Biophys, Houston, TX 77030 USA. NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Jing, NJ (reprint author), Baylor Coll Med, Dept Mol Physiol & Biophys, Houston, TX 77030 USA. RI Marchand, Christophe/D-8559-2016 FU NIGMS NIH HHS [GM60153] NR 40 TC 85 Z9 88 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 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 14 PY 2000 VL 275 IS 28 BP 21460 EP 21467 DI 10.1074/jbc.M001436200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 335EM UT WOS:000088230600068 PM 10801812 ER PT J AU Fukuhara, S Marinissen, MJ Chiariello, M Gutkind, JS AF Fukuhara, S Marinissen, MJ Chiariello, M Gutkind, JS TI Signaling from G protein-coupled receptors to ERK5/big MAPK 1 involves G alpha(q) and G alpha(12/13) families of heterotrimeric G proteins - Evidence for the existence of a novel Ras and Rho-independent pathway SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GTP-BINDING PROTEINS; BETA-GAMMA-SUBUNITS; JUN NH2-TERMINAL KINASE; N-TERMINAL KINASE; TYROSINE KINASE; TRANSCRIPTIONAL ACTIVATION; DEPENDENT ACTIVATION; GENE-EXPRESSION; INVOLVEMENT; THROMBIN AB The regulation of gene expression by cell surface receptors often involves the stimulation of signaling pathways including one or more members of the MAPK superfamily of serine-threonine kinases. Upon their activation in the cytosol, MAPKs can translocate to the nucleus and affect the activity of a variety of transcription factors. Recently, it has been observed that a novel member of the MAPK superfamily, ERK5, can be potently activated by transforming G protein-coupled receptors (GPCRs) and that ERK5 participates in the regulation of c-jun expression through the activation of MEF2 transcription factors. How cell surface receptors, including GPCRs, stimulate ERK5 is still poorly understood. In this study, we have used transiently transfected COS-7 cells to begin delineating the biochemical route linking GPCRs to ERK5. We show that receptors that can couple to the Gq and G(12/13) families of heterotrimeric G proteins, mi and thrombin receptors, respectively, but not those coupled to G(i), such as m2 receptors, are able to regulate the activity of ERK5. To investigate which heterotrimeric G proteins signal to ERK5, we used a chimeric system by which G alpha(q)- and G alpha(13)-mediated signaling pathways can be conditionally activated upon ligand stimulation. Using this system, as well as the expression of activated forms of G protein subunits, we show that the G alpha(q) and G alpha(12/13) families of heterotrimeric G proteins, but not the G alpha(i), G alpha(s), and beta gamma subunits, are able to regulate ERK5. Furthermore, we provide evidence that the stimulation of ERK5 by GPCRs involves a novel signaling pathway, which is distinct from those regulated by Ras and Rho GTPases. C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Gutkind, JS (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, Rm 212,Bldg 30,30 Convent Dr, Bethesda, MD 20892 USA. RI Chiariello, Mario/O-3642-2014 OI Chiariello, Mario/0000-0001-8434-5177 NR 48 TC 70 Z9 74 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 14 PY 2000 VL 275 IS 28 BP 21730 EP 21736 DI 10.1074/jbc.M002410200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 335EM UT WOS:000088230600101 PM 10781600 ER PT J AU Zhang, SJ Cheng, HP Zhou, YY Wang, DJ Zhu, WZ Ziman, B Spurgoen, H Lefkowitz, RJ Lakatta, EG Koch, WJ Xiao, RP AF Zhang, SJ Cheng, HP Zhou, YY Wang, DJ Zhu, WZ Ziman, B Spurgoen, H Lefkowitz, RJ Lakatta, EG Koch, WJ Xiao, RP TI Inhibition of spontaneous beta(2)-adrenergic activation rescues beta(1)-adrenergic contractile response in cardiomyocytes overexpressing beta(2)-adrenoceptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ADRENERGIC-RECEPTOR KINASE; PROTEIN-COUPLED RECEPTORS; RAT-HEART; PHOSPHOLAMBAN PHOSPHORYLATION; BETA-ADRENOCEPTOR; CARDIAC MYOCYTES; TRANSGENIC MICE; CA2+ CURRENT; DESENSITIZATION; EXPRESSION AB Cardiac-specific overexpression of the human beta(2)-adrenergic receptor (AR) in transgenic mice (TG4) enhances basal cardiac function due to ligand-independent spontaneous beta(2)-AR activation. However, agonist-mediated stimulation of either beta(1)-AR or beta(2)-AR fails to further enhance contractility in TG4 ventricular myocytes. Although the lack of beta(2)-AR response has been ascribed to an efficient coupling of the receptor to pertussis toxin-sensitive G(i) proteins in addition to G(s), the contractile response to beta(1)-AR stimulation by norepinephrine and an alpha(1)-adrenergic antagonist prazosin is not restored by pertussis toxin treatment despite a G(i) protein elevation of 1.7-fold in TG4 hearts. Since beta-adrenergic receptor kinase, beta ARK1, activity remains unaltered, the unresponsiveness of beta(1)-AR is not caused by beta ARK1-mediated receptor desensitization. In contrast, pre-incubation of cells with anti-adrenergic reagents such as muscarinic receptor agonist, carbachol (10(-5) M), or a beta(2)-AR inverse agonist, ICI 118,551 (5 x 10(-7) M), to abolish spontaneous beta(2)-AR signaling, both reduce the base-line cAMP and contractility and, surprisingly, restore the beta(1)-AR contractile response. The "rescued" contractile response is completely reversed by a beta(1)-AR antagonist, CGP 20712A. Furthermore, these results from the transgenic animals are corroborated by in vitro acute gene manipulation in cultured wild type adult mouse ventricular myocytes. Adenovirus-directed overexpression of the human beta(2)-AR results in elevated baseline cAMP and contraction associated with a marked attenuation of beta(1)-AR response; carbachol pretreatment fully revives the diminished beta(1)-AR contractile response. Thus, we conclude that constitutive beta(2)-AR activation induces a heterologous desensitization of beta(1)-ARs independent of beta ARK1 and G(i) proteins; suppression of the constitutive beta(2)-AR signaling by either a beta(2)-AR inverse agonist or stimulation of the muscarinic receptor rescues the beta(1)-ARs from desensitization, permitting agonist-induced contractile response. C1 NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA. Duke Univ, Med Ctr, Dept Med, Durham, NC 27710 USA. Duke Univ, Med Ctr, Howard Hughes Med Inst, Durham, NC 27710 USA. RP Xiao, RP (reprint author), NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 36 TC 19 Z9 20 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 14 PY 2000 VL 275 IS 28 BP 21773 EP 21779 DI 10.1074/jbc.M909484199 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 335EM UT WOS:000088230600107 PM 10787424 ER PT J AU Monory, K Bryant, SD Kertesz, I Balboni, G Guerrini, R Toth, G Salvadori, S Lazarus, LH Borsodi, A AF Monory, K Bryant, SD Kertesz, I Balboni, G Guerrini, R Toth, G Salvadori, S Lazarus, LH Borsodi, A TI Characterization of N,N(Me)(2)-Dmt-Tic-OH, a delta selective opioid dipeptide antagonist SO NEUROREPORT LA English DT Article DE antagonist; delta opioid receptor; [S-35]GTP gamma S binding; peptide; radioligand binding ID DMT-TIC PHARMACOPHORE; RECEPTOR; BINDING; DIKETOPIPERAZINE; MEMBRANES; AFFINITY; PEPTIDES; ACID AB N,N(Me)(2)-Dimethyl-tyrosine-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid-OH (N,N(Me)(2)-Dmt-Tic-OH) is a very selective delta opioid dipeptide with elevated antagonist activity. We have radiolabelled this compound by catalytic tritiation of the N,N(Me)(2)-Dmt(3',5'-I-2)-Tic-OH precursor. The ligand labelled rat brain membranes with a K-d value of 0.42 nM and a B-max of 63.12 fmol/mg protein. The new tritiated ligand showed high affinity for the delta opioid receptor whereas its binding at mu and kappa opioid receptors was weak. N,N(Me)(2)-Dmt-Tic-OH was able to inhibit the agonist-stimulated binding of the non-hydrolysable GTP analogue [S-35]GTP gamma S, thus attenuating the activation of G proteins via opioid receptors. This simple opioid dipeptide in both normal and labelled form may serve as a useful tool to study delta opioid receptors in vitro and in vivo. NeuroReport 11:2083-2086 (C) 2000 Lippincott Williams & Wilkins. C1 Hungarian Acad Sci, Biol Res Ctr, Inst Biochem, H-6701 Szeged, Hungary. Hungarian Acad Sci, Biol Res Ctr, Isotope Lab, H-6701 Szeged, Hungary. Univ Ferrara, Dipartimento Sci Farmaceut, I-44100 Ferrara, Italy. Univ Ferrara, Ctr Biotechnol, I-44100 Ferrara, Italy. NIEHS, LCBRA, Res Triangle Pk, NC 27709 USA. RP Borsodi, A (reprint author), Hungarian Acad Sci, Biol Res Ctr, Inst Biochem, POB 521, H-6701 Szeged, Hungary. OI Guerrini, Remo/0000-0002-7619-0918; SALVADORI, Severo/0000-0002-8224-2358 NR 19 TC 3 Z9 3 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-4965 J9 NEUROREPORT JI Neuroreport PD JUL 14 PY 2000 VL 11 IS 10 BP 2083 EP 2086 DI 10.1097/00001756-200007140-00005 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 336BU UT WOS:000088282700005 PM 10923648 ER PT J AU Kanduri, C Pant, V Loukinov, D Pugacheva, E Qi, CF Wolffe, A Ohlsson, R Lobanenkov, VV AF Kanduri, C Pant, V Loukinov, D Pugacheva, E Qi, CF Wolffe, A Ohlsson, R Lobanenkov, VV TI Functional association of CTCF with the insulator upstream of the H19 gene is parent of origin-specific and methylation-sensitive SO CURRENT BIOLOGY LA English DT Article ID DNA-SEQUENCE; EXPRESSION; DELETION; IGF2 AB in mammals, a subset of genes inherit gametic marks that establish parent of origin-dependent expression patterns in the soma ([1] and references therein). The currently most extensively studied examples of this phenomenon, termed genomic imprinting, are the physically linked Igf2 (insulin like growth factor II) and H19 genes, which are expressed mono-allelically from opposite parental alleles [1,2]. The repressed status of the maternal Igf2 allele is due to cis elements that prevent the HIS enhancers [3] from accessing the Igf2 promoters on the maternal chromosome [4,5], A differentially methylated domain (DMD) in the 5' flank of H19 is maintained paternally methylated and maternally unmethylated [6,7], We show here by gel-shift and chromatin immunopurification analyses that binding of the highly conserved multivalent factor CTCF ([8,9] and references therein) to the His DMD is methylation-sensitive and parent of origin-dependent, Selectively mutating CTCF-contacting nucleotides, which were identified by methylation interference within the extended binding sites initially revealed by nuclease footprinting, abrogated the His DMD enhancer blocking property. These observations suggest that molecular mechanisms of genomic Imprinting may use an unusual ability of CTCF to interact with a diverse spectrum of variant target sites, some of which include CpGs that are responsible for methylation-sensitive CTCF binding in vitro and in vivo. C1 Sangamo Biosci Inc, Point Richmond Tech Ctr, Richmond, CA 94804 USA. Uppsala Univ, Dept Genet & Dev, S-75236 Uppsala, Sweden. NIAID, Mol Pathol Sect, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. RP Lobanenkov, VV (reprint author), Sangamo Biosci Inc, Point Richmond Tech Ctr, 501 Canal Blvd,Suite A100, Richmond, CA 94804 USA. OI Lobanenkov, Victor/0000-0001-6665-3635 NR 10 TC 320 Z9 327 U1 1 U2 5 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD JUL 13 PY 2000 VL 10 IS 14 BP 853 EP 856 DI 10.1016/S0960-9822(00)00597-2 PG 4 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 348HJ UT WOS:000088979200019 PM 10899010 ER PT J AU Pettit, GR Grealish, MP Herald, DL Boyd, MR Hamel, E Pettit, RK AF Pettit, GR Grealish, MP Herald, DL Boyd, MR Hamel, E Pettit, RK TI Antineoplastic agents. 443. Synthesis of the cancer cell growth inhibitor hydroxyphenstatin and its sodium diphosphate prodrug SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID COMBRETASTATIN A-4 PRODRUG; GRIGNARD-REAGENTS; KETONES; TUBULIN; CHLORIDES; PHOSPHATE AB A structure-activity relationship (SAR) study of the South African willow tree (Combretum caffrum) antineoplastic constituent combretastatin A-4 (3b) led to the discovery of a potent cancer cell growth inhibitor designated phenstatin (5a). This benzophenone derivative of combretastatin A-4 showed remarkable antineoplastic activity, and the benzophenone derivative of combretastatin A-1 was therefore synthesized. The benzophenone, designated hydroxyphenstatin (6a), was synthesized by coupling of a protected bromobenzene and a benzaldehyde to give the benzhydrol with subsequent oxidation to the ketone. Hydroxyphenstatin was converted to the sodium phosphate prodrug (6e) by a dibenzyl phosphite phosphorylation and subsequent benzyl cleavage (6a --> 6d --> 6e). While hydroxyphenstatin (6a) was a potent inhibitor of tubulin polymerization with activity comparable to that of combretastatin A-1 (3a), the phosphorylated derivative (6e) was inactive. C1 Arizona State Univ, Canc Res Inst, Tempe, AZ 85287 USA. Arizona State Univ, Dept Chem & Biochem, Tempe, AZ 85287 USA. NCI, Lab Drug Discovery Res & Dev, DTP, DCTD,Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, Screening Technol Branch, DTP, DCTD,Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Pettit, GR (reprint author), Arizona State Univ, Canc Res Inst, POB 872404, Tempe, AZ 85287 USA. NR 32 TC 74 Z9 75 U1 1 U2 6 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 13 PY 2000 VL 43 IS 14 BP 2731 EP 2737 DI 10.1021/jm000045a PG 7 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 333XY UT WOS:000088157100010 PM 10893310 ER PT J AU Gottlieb, M McGovern, V Goodwin, P Hoffman, S Oduola, A AF Gottlieb, M McGovern, V Goodwin, P Hoffman, S Oduola, A TI Please don't downgrade the sequencers' role ... SO NATURE LA English DT Letter C1 NIAID, Bethesda, MD 20892 USA. Burroughs Wellcome Fund, Res Triangle Pk, NC 27709 USA. Wellcome Trust, London NW1 2BE, England. USN, Med Res Ctr, Silver Spring, MD 20910 USA. WHO, Special Programme Res & Training Trop Dis, World Bank, UNDP, CH-1211 Geneva 27, Switzerland. RP Gottlieb, M (reprint author), NIAID, 6700-B Rockledge Dr, Bethesda, MD 20892 USA. NR 1 TC 3 Z9 3 U1 0 U2 0 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 13 PY 2000 VL 406 IS 6792 BP 121 EP 122 DI 10.1038/35018270 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335BA UT WOS:000088221100014 PM 10910329 ER PT J AU Hallett, M AF Hallett, M TI Transcranial magnetic stimulation and the human brain SO NATURE LA English DT Article ID MOTOR CORTEX EXCITABILITY; VISUAL-CORTEX; PARKINSONS-DISEASE; CORTICAL OUTPUTS; CEREBRAL-CORTEX; READING HAND; REORGANIZATION; DEPRESSION; MODULATION; ORGANIZATION AB Transcranial magnetic stimulation (TMS) is rapidly developing as a powerful, non-invasive tool for studying the human brain. A pulsed magnetic field creates current flow in the brain and can temporarily excite or inhibit specific areas. TMS of motor cortex can produce a muscle twitch or block movement; TMS of occipital cortex can produce visual phosphenes or scotomas. TMS can also alter the functioning of the brain beyond the time of stimulation, offering potential for therapy. C1 NINDS, Human Motor Control Sect, NIH, Bethesda, MD 20892 USA. RP Hallett, M (reprint author), NINDS, Human Motor Control Sect, NIH, Bldg 10,Room 5N226,10 Ctr Dr,MSC 1428, Bethesda, MD 20892 USA. NR 41 TC 668 Z9 695 U1 17 U2 106 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 13 PY 2000 VL 406 IS 6792 BP 147 EP 150 DI 10.1038/35018000 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335BA UT WOS:000088221100036 PM 10910346 ER PT J AU Pearson, M Carbone, R Sebastiani, C Cioce, M Fagioli, M Saito, S Higashimoto, Y Appella, E Minucci, S Pandolfi, PP Pelicci, PG AF Pearson, M Carbone, R Sebastiani, C Cioce, M Fagioli, M Saito, S Higashimoto, Y Appella, E Minucci, S Pandolfi, PP Pelicci, PG TI PML regulates p53 acetylation and premature senescence induced by oncogenic Ras SO NATURE LA English DT Article ID CREB BINDING-PROTEIN; WILD-TYPE P53; PROMYELOCYTIC LEUKEMIA; GENE AMPLIFICATION; DNA-DAMAGE; IN-VITRO; CELLS; MODULATION; GROWTH; TRANSFORMATION AB The tumour suppressor p53 induces cellular senescence in response to oncogenic signals(1). p53 activity is modulated by protein stability and post-translational modification, including phosphorylation and acetylation(2). The mechanism of p53 activation by oncogenes remains largely unknown. Here we report that the tumour suppressor PML regulates the p53 response to oncogenic signals. We found that oncogenic pas upregulates PML expression, and overexpression of PML induces senescence in a p53-dependent manner. p53 is acetylated at lysine 382 upon Ras expression, an event that is essential for its biological function. pas induces re-localization of p53 and the CBP acetyltransferase within the PML nuclear bodies and induces the formation of a trimeric p53-PML-CBP complex. Lastly, Ras-induced p53 acetylation, p53-CBP complex stabilization and senescence are lost in PML-/- fibroblasts. Our data establish a link between PML and p53 and indicate that integrity of the PML bodies is required for p53 acetylation and senescence upon oncogene expression. C1 European Inst Oncol, Dept Expt Oncol, I-20141 Milan, Italy. Univ Perugia, Ist Med Interna & Sci Oncol, I-06100 Perugia, Italy. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. Cornell Univ, Mem Sloan Kettering Canc Ctr, Dept Human Genet, New York, NY 10021 USA. Cornell Univ, Mem Sloan Kettering Canc Ctr, Program Mol Biol, New York, NY 10021 USA. RP Pelicci, PG (reprint author), European Inst Oncol, Dept Expt Oncol, I-20141 Milan, Italy. RI Minucci, Saverio/J-9669-2012; Tang, Amy/L-3226-2016 OI Tang, Amy/0000-0002-5772-2878 NR 30 TC 548 Z9 563 U1 2 U2 17 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUL 13 PY 2000 VL 406 IS 6792 BP 207 EP 210 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 335BA UT WOS:000088221100054 PM 10910364 ER PT J AU Hoover, RN AF Hoover, RN TI Cancer - Nature, nurture, or both SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material ID BREAST-CANCER; WOMEN C1 NCI, Rockville, MD 20892 USA. RP Hoover, RN (reprint author), NCI, Rockville, MD 20892 USA. NR 9 TC 57 Z9 57 U1 0 U2 4 PU MASSACHUSETTS MEDICAL SOC 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 13 PY 2000 VL 343 IS 2 BP 135 EP 136 DI 10.1056/NEJM200007133430210 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 333EP UT WOS:000088116500010 PM 10891523 ER PT J AU Davey, RT Murphy, RL Graziano, FM Boswell, SL Pavia, AT Cancio, M Nadler, JP Chaitt, DG Dewar, RL Sahner, DK Duliege, AM Capra, WB Leong, WP Giedlin, MA Lane, HC Kahn, JO AF Davey, RT Murphy, RL Graziano, FM Boswell, SL Pavia, AT Cancio, M Nadler, JP Chaitt, DG Dewar, RL Sahner, DK Duliege, AM Capra, WB Leong, WP Giedlin, MA Lane, HC Kahn, JO TI Immunologic and virologic effects of subcutaneous interleukin 2 in combination with antiretroviral therapy - A randomized controlled trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; INTRAVENOUS INTERLEUKIN-2; INFECTION; TYPE-1; ZIDOVUDINE; PLASMA; HAART AB Context While interleukin 2 (IL-2) is capable of inducing a marked expansion of the CD4 T-lymphocyte pool, limited data exist on whether IL-2 treatment can add significantly to the immunologic and virologic effects of potent antiretroviral therapy (ART). Objective To determine the rate and magnitude of CD4 cell recovery and viral suppression when using a combination therapy of IL-2 and ART com pared with ART alone. Design and Setting Randomized, controlled multicenter trial conducted from April 1996 through April 1998 at 8 clinical sites in the United States. Patients Eighty-two adult outpatients who were infected with human immunodeficiency virus (HIV) and had baseline CD4 cell counts of 200 x 10(6)/L to 500 x 10(6)/L and baseline RNA levels of fewer than 10 000 copies/mL were randomized; 78 completed the study. Interventions Thirty-nine patients were randomly assigned to receive a combination therapy of subcutaneous IL-2 (administered in 5-day courses every 8 weeks at a starting dosage of 7.5 mIU twice per day) and ART; 43 were to receive ART therapy alone. Main Outcome Measures Interleukin 2 safety and differential effects on CD4 cell counts, CD4 cell percentages, and plasma HIV RNA levels. Results The mean (SD) percentage increase in CD4 cell counts at 1 year for patients who received IL-2 was 112% (113%) compared with 18% (35%) in recipients of ART alone (P<.001). Both groups had, mean (SD) increases in CD4 cell percentage: from 20.4% (6.3%) to 32.3% (12.4%) for the combination therapy group compared with 20.4% (5.1%) to 23.0% (7.2%) for recipients of ART alone (P<.001). Using a sensitive viral RNA assay, mean viral load changes were -0.28 and 0.09 log(10) copies for IL-2 recipients and control patients, respectively (P=.03). Twenty (67%) of 30 evaluable patients receiving IL-2 achieved final viral loads of fewer than 50 copies/mL compared with 13 (36%) of 36 control patients (P=.02). Toxic effects were common among patients who received IL-2 and were managed with antipyretics, hydration, rest, and dosage reduction as needed. Conclusions Intermittent therapy with IL-2 and ART produced a substantially greater increase in CD4 cells and was associated with a larger decrease in viral load than ART alone. Clinical end-point trials will be necessary to determine whether the enhanced viral suppression and CD4 cell increases associated with IL-2 therapy will translate into improved clinical outcomes. C1 NIAID, NIH, Bethesda, MD 20892 USA. NIH, Dept Crit Care Med, Bethesda, MD 20892 USA. Northwestern Univ, Div Infect Dis, Chicago, IL 60611 USA. Univ Wisconsin, Dept Med, Madison, WI USA. Fenway Community Hlth Ctr, Boston, MA USA. Univ Utah, Div Pediat Infect Dis, Salt Lake City, UT USA. St Josephs Hosp, Tampa, FL USA. Univ S Florida, Div Infect Dis, Tampa, FL USA. Sci Applicat Int Corp, Frederick, MD USA. Chiron Corp, Emeryville, CA 94608 USA. San Francisco Gen Hosp, San Francisco, CA 94110 USA. Univ Calif San Francisco, Ctr AIDS Res, San Francisco, CA 94143 USA. RP Davey, RT (reprint author), NIAID, NIH, Bldg 10,Room 11C-103, Bethesda, MD 20892 USA. OI Murphy, Robert/0000-0003-3936-2052 FU PHPPO CDC HHS [PH30 MH59037] NR 26 TC 142 Z9 144 U1 0 U2 7 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 12 PY 2000 VL 284 IS 2 BP 183 EP 189 DI 10.1001/jama.284.2.183 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 329XE UT WOS:000087933100022 PM 10889591 ER PT J AU Biggar, RJ Frisch, M Goedert, JJ AF Biggar, RJ Frisch, M Goedert, JJ CA AIDS-Canc Match Registry Study Grp TI Risk of cancer in children with AIDS SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID KAPOSIS-SARCOMA; HIV-INFECTION; LYMPHOMA; EPIDEMIOLOGY; PEOPLE AB Context Population-based data on cancers associated with acquired immunodeficiency syndrome (AIDS) in children are lacking. Objective To determine risk of pediatric AIDS-associated cancers. Design, Setting, and Participants Using records from 11 locations in the United States for varying periods between 1978 and 1996, we linked data for children aged 14 years and younger at AIDS diagnosis to local cancer registry data. Main Outcome Measures Cancer frequency and, in the 2-year post-AIDS onset period, cancer incidence and relative risk (RR; measured as standardized incidence ratio), by cancer type. Results Among 4954 children with AIDS, 124 (2.5%) were identified as having cancer before, at, or after AIDS onset, including 100 cases of non-Hodgkin lymphoma (NHL), 8 of Kaposi sarcoma (KS), 4 of leiomyosarcoma, and 2 of Hodgkin disease; there were 10 other or unspecified cancers. Expected numbers for all cancers identified in the study sample, based on population rates (using area-specific registry data), were less than 1. In the first 2 years after AIDS diagnosis (5485 person-years), NHL incidence was 510 per 100 000 person-years (RR, 651; 95% confidence interval [CI], 432-941). Median time for developing NHL after AIDS diagnosis was 14 months (range, 3-107 months). The most common type of NHL was Burkitt lymphoma. However, the risk of primary brain lymphoma (91 per 100 000 person-years) was especially high (RR, 7143; 95% CI, 2321-16692), and 4 cases were diagnosed more than 2 years (range, 37-98 months) after AIDS onset. Leiomyosarcomas also tended to occur several years after AIDS onset, with 3 of the 4 cases occurring 33 to 76 months after AIDS diagnosis, whereas KS was reported only at or within 2 years of AIDS diagnosis. Hodgkin disease risk was also significantly increased (RR, 62; 95% CI, 2-342). Conclusions The spectrum of AIDS-associated pediatric cancers resembled that seen in adults, with the addition of leiomyosarcoma. Both primary brain lymphomas and leiomyosarcomas tended to occur in children surviving several years after AIDS onset. Because the expected numbers of these cancers in this population were less than 1 and because of the small numbers of some types of observed cancers, the RR estimates are imprecise and caution is warranted in their interpretation. C1 NCI, VEB, Rockville, MD 20852 USA. Statens Serum Inst, Danish Epidemiol Sci Ctr, Dept Epidemiol Res, DK-2300 Copenhagen, Denmark. RP Biggar, RJ (reprint author), NCI, VEB, 6120 Execut Blvd,Room 8014, Rockville, MD 20852 USA. RI Frisch, Morten/E-9206-2016 OI Frisch, Morten/0000-0002-3864-8860 NR 16 TC 78 Z9 85 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 12 PY 2000 VL 284 IS 2 BP 205 EP 209 DI 10.1001/jama.284.2.205 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 329XE UT WOS:000087933100025 PM 10889594 ER PT J AU D'Souza, MP Cairns, JS Plaeger, SF AF D'Souza, MP Cairns, JS Plaeger, SF TI Current evidence and future directions for targeting HIV entry - Therapeutic and prophylactic strategies SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; CHEMOKINE RECEPTOR CXCR4; SMALL-MOLECULE INHIBITOR; SYNCYTIUM-INDUCING PHENOTYPE; CANDIDATE AIDS VACCINES; INFECTION IN-VIVO; DISEASE PROGRESSION; ENVELOPE GLYCOPROTEIN; COILED-COIL; NEUTRALIZING ANTIBODIES AB Great strides have been made in developing potent antiretroviral regimens that block human immunodeficiency virus (HIV) transcription and assembly. Despite these therapeutic advances, problems of drug resistance, latent viral reservoirs, and drug-induced toxic effects that compromise effective viral central point to the need for new classes of anti-HIV drugs with different modes of action. One promising approach involves blocking HIV entry into human cells, a complex process that involves multiple protein interactions. The process of HIV entry begins with binding of the viral envelope glycoprotein to both the CD4 receptor and one of several chemokine receptors and ends with fusion of viral and cell membranes. Conceptually, there are 3 steps in the HIV entry process that could serve as therapeutic targets: binding of the viral envelope glycoprotein with the CD4 receptor, binding of the envelope-CD4 complex to chemokine receptors, and fusion of the viral and cell membranes. Preclinical and clinical assessment of these entry inhibitors is ongoing and will determine if they possess properties required for drug licensure. Moreover, the worldwide epidemic is largely occurring in developing countries that cannot afford these drugs: a prophylactic vaccine is necessary and urgent. New knowledge of the HIV-envelope glycoprotein has also provided insight into possibilities for the design of novel HIV vaccines. C1 NIAID, Vaccine Clin Res Branch, Div AIDS, NIH, Bethesda, MD 20892 USA. NIAID, Branch Targeted Intervent, Div AIDS, NIH, Bethesda, MD 20892 USA. NIAID, Branch Pathogenesis & Basic Sci, Div AIDS, NIH, Bethesda, MD 20892 USA. RP D'Souza, MP (reprint author), NIAID, Vaccine Clin Res Branch, Div AIDS, NIH, Room 4152,6700-B Rockledge Dr, Bethesda, MD 20892 USA. NR 99 TC 77 Z9 78 U1 0 U2 3 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 12 PY 2000 VL 284 IS 2 BP 215 EP 222 DI 10.1001/jama.284.2.215 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 329XE UT WOS:000087933100027 PM 10889596 ER PT J AU Klein, HG AF Klein, HG TI Will blood transfusion ever be safe enough? SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Editorial Material ID VIRUS; RISK C1 NIH, Dept Transfus Med, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Klein, HG (reprint author), NIH, Dept Transfus Med, Warren G Magnuson Clin Ctr, Bldg 10,Room 1C711,MSC-1184, Bethesda, MD 20892 USA. NR 22 TC 30 Z9 30 U1 0 U2 2 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 12 PY 2000 VL 284 IS 2 BP 238 EP 240 DI 10.1001/jama.284.2.238 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 329XE UT WOS:000087933100031 PM 10889600 ER PT J AU Jaretzki, A Barohn, RJ Ernstoff, RM Kaminski, HJ Keesey, JC Penn, AS Sanders, DB AF Jaretzki, A Barohn, RJ Ernstoff, RM Kaminski, HJ Keesey, JC Penn, AS Sanders, DB CA Myasthenia Gravis Fdn Amer TI Myasthenia gravis - Recommendations for clinical research standards SO NEUROLOGY LA English DT Review ID LONG-TERM CORRELATION; THYMIC CARCINOMA; DOUBLE-BLIND; THYMECTOMY; THYMOMA; DISEASE; AZATHIOPRINE; SEVERITY; QUALITY; PATIENT C1 Myasthenia Gravis Fdn Amer Inc, Chicago, IL 60602 USA. Columbia Presbyterian Med Ctr, Dept Surg, New York, NY 10032 USA. Univ Texas, SW Med Ctr, Dept Neurol, Dallas, TX USA. William Beaumont Hosp, Serv Neurol, Royal Oak, MI USA. Univ Hosp Cleveland, Dept Vet Affairs Med Ctr, Case Western Reserve Univ, Dept Neurol, Cleveland, OH 44106 USA. Univ Hosp Cleveland, Dept Vet Affairs Med Ctr, Case Western Reserve Univ, Dept Neurosci, Cleveland, OH 44106 USA. Univ Calif Los Angeles, Dept Neurol, Los Angeles, CA 90024 USA. Natl Inst Neurol Disorders & Stroke, NIH, Bethesda, MD USA. Duke Univ, Med Ctr, Dept Med, Durham, NC USA. RP Jaretzki, A (reprint author), Myasthenia Gravis Fdn Amer Inc, 123 W Madison,Suite 800, Chicago, IL 60602 USA. NR 51 TC 501 Z9 526 U1 2 U2 25 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUL 12 PY 2000 VL 55 IS 1 BP 16 EP 23 PG 8 WC Clinical Neurology SC Neurosciences & Neurology GA 332NV UT WOS:000088081100005 PM 10891897 ER PT J AU Moffat, SD Szekely, CA Zonderman, AB Kabani, NJ Resnick, SM AF Moffat, SD Szekely, CA Zonderman, AB Kabani, NJ Resnick, SM TI Longitudinal change in hippocampal volume as a function of apolipoprotein E genotype SO NEUROLOGY LA English DT Article ID E EPSILON-4 ALLELE; ALZHEIMERS-DISEASE AB The epsilon 4 allele of the apolipoprotein E (APOE) gene confers an increased risk for the development of AD. The authors compared longitudinal rates of change in hippocampal volume as a function of APOE genotype in nondemented elderly individuals. Rate of volumetric loss was significantly greater among epsilon 4+ compared with epsilon 4- individuals. These results indicate that individuals positive for the APOE epsilon 4 allele may show a greater rate of hippocampal atrophy than their epsilon 4- counterparts, even in the absence of a diagnosis of AD. C1 NIA, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Montreal Neurol Hosp & Inst, McConnell Brain Imaging Ctr, Montreal, PQ H3A 2B4, Canada. RP Resnick, SM (reprint author), NIA, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Szekely, Christine/C-1342-2009; OI Zonderman, Alan B/0000-0002-6523-4778 NR 10 TC 114 Z9 117 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUL 12 PY 2000 VL 55 IS 1 BP 134 EP 136 PG 3 WC Clinical Neurology SC Neurosciences & Neurology GA 332NV UT WOS:000088081100030 PM 10891924 ER PT J AU Backgren, C Hummer, G Wikstrom, M Puustinen, A AF Backgren, C Hummer, G Wikstrom, M Puustinen, A TI Proton translocation by cytochrome c oxidase can take place without the conserved glutamic acid in subunit I SO BIOCHEMISTRY LA English DT Article ID HEME-COPPER OXIDASES; SENSORY RHODOPSIN-I; PARACOCCUS-DENITRIFICANS; ESCHERICHIA-COLI; RHODOBACTER-SPHAEROIDES; STRUCTURAL-CHANGES; UBIQUINOL OXIDASE; OXYGEN; BO(3); RESOLUTION AB A glutamic acid residue in subunit I of the heme-copper oxidases is highly conserved and has been directly implicated in the O-2 reduction and proton-pumping mechanisms of these respiratory enzymes. Its mutation to residues other than aspartic acid dramatically inhibits activity, and proton translocation is lost. However, this glutamic acid is replaced by a nonacidic residue in some structurally distant members of the heme-copper oxidases, which have a tyrosine residue in the vicinity. Here, using cytochrome c oxidase from Paracoccus denitrificans, we show that replacement of the glutamic acid and a conserved glycine nearby lowers the catalytic activity to <0.1% of the wild-type value. But if, in addition, a phenylalanine that lies close in the structure is changed to tyrosine, the activity rises more than 100-fold and proton translocation is restored. Molecular dynamics simulations suggest that the tyrosine can support a transient array of water molecules that may be essential for proton transfer in the heme-copper oxidases. Surprisingly, the glutamic acid is thus not indispensable, which puts important constraints on the catalytic mechanism of these enzymes. C1 Univ Helsinki, Inst Biomed Sci, Dept Med Chem, Helsinki Bioenerget Grp, FIN-00014 Helsinki, Finland. Univ Helsinki, Bioctr Helsinki, FIN-00014 Helsinki, Finland. NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Puustinen, A (reprint author), Univ Helsinki, Inst Biomed Sci, Dept Med Chem, Helsinki Bioenerget Grp, POB 8, FIN-00014 Helsinki, Finland. RI Wikstrom, Marten/A-4403-2008; Hummer, Gerhard/A-2546-2013 OI Hummer, Gerhard/0000-0001-7768-746X NR 30 TC 64 Z9 64 U1 0 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 11 PY 2000 VL 39 IS 27 BP 7863 EP 7867 DI 10.1021/bi000806b PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 333KA UT WOS:000088128400004 PM 10891065 ER PT J AU Driscoll, WJ Konig, S Fales, HM Pannell, LK Eipper, BA Mueller, GP AF Driscoll, WJ Konig, S Fales, HM Pannell, LK Eipper, BA Mueller, GP TI Peptidylglycine-alpha-hydroxylating monooxygenase generates two hydroxylated products from its mechanism-based suicide substrate, 4-phenyl-3-butenoic acid SO BIOCHEMISTRY LA English DT Article ID DOPAMINE BETA-HYDROXYLASE; ENZYME-BOUND COPPER; C-TERMINAL AMIDES; AMIDATING MONOOXYGENASE; PEPTIDE AMIDATION; HYDROXYGLYCINE INTERMEDIATE; NEUROPEPTIDE AMIDATION; INACTIVATION; CELLS; STEREOCHEMISTRY AB The bifunctional enzyme peptidylglycine-alpha-amidating monooxygenase mediates the conversion of C-terminal glycine-extended peptides to their active alpha-amidated products. Peptidylglycine-alpha-hydroxylating monooxygenase (PHM, EC 1.14. 17.3) catalyzes the first reaction in this two-step process. The olefinic compound 4-phenyl-3-butenoic acid (PBA) is the most potent irreversible, mechanism-based PHM inactivator known. While the details of the inhibitory action of PEA on PHM remain undefined, covalent modification of the protein has been proposed as the underlying mechanism. We report here that, in the process of inactivating PHM, PEA itself serves as a substrate without covalently labeling the enzyme. Approximately 100 molecules of PEA are metabolized per molecule of PHM inactivated, under saturating conditions. The metabolism of PEA by PHM generates two hydroxylated products, 2-hydroxy-4-phenyl-3-butenoic acid and its allylic isomer, 4-hydroxy-4-phenyl-2-butenoic acid. While one enantiomer for each product is significantly favored in the reaction, both are produced. From these observations, we conclude that hydroxylated PEA products are formed by a delocalized free radical mechanism and that the lack of absolute stereospecificity indicates significant freedom of movement within the catalytic site, The ability of PHM to metabolize PBA suggests that the physiological functions of PHM may include the hydroxylation of substrates other than those containing terminal glycines. C1 Uniformed Serv Univ Hlth Sci, Dept Physiol, F Edward Hebert Sch Med, Bethesda, MD 20814 USA. NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA. RP Driscoll, WJ (reprint author), Uniformed Serv Univ Hlth Sci, Dept Physiol, F Edward Hebert Sch Med, 4301 Jones Bridge Rd, Bethesda, MD 20814 USA. RI Konig, Simone/B-6504-2008 OI Konig, Simone/0000-0003-0672-7246 FU NIDDK NIH HHS [DK32949]; NINDS NIH HHS [NS34173]; PHS HHS [R07644] NR 44 TC 11 Z9 11 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 11 PY 2000 VL 39 IS 27 BP 8007 EP 8016 DI 10.1021/bi0002380 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 333KA UT WOS:000088128400021 PM 10891082 ER PT J AU Knatterud, GL Rosenberg, Y Campeau, L Geller, NL Hunninghake, DB Forman, SA Forrester, JS Gobel, FL Herd, JA Hickey, A Hoogwerf, BJ Terrin, ML White, C AF Knatterud, GL Rosenberg, Y Campeau, L Geller, NL Hunninghake, DB Forman, SA Forrester, JS Gobel, FL Herd, JA Hickey, A Hoogwerf, BJ Terrin, ML White, C CA Post CABG Investigators TI Long-term effects on clinical outcomes of aggressive lowering of low-density lipoprotein cholesterol levels and low-dose anticoagulation in the post coronary artery bypass graft trial SO CIRCULATION LA English DT Article DE bypass; grafting; anticoagulation; lipids; follow-up studies ID PROGNOSTIC-SIGNIFICANCE; MYOCARDIAL-INFARCTION; EVENTS; ATHEROSCLEROSIS; PROGRESSION AB Background-The Post Coronary Artery Bypass Graft Trial, designed to compare the effects of 2 lipid-lowering regimens and low-dose anticoagulation versus placebo on progression of atherosclerosis in saphenous vein grafts of patients who had had CABG surgery, demonstrated that aggressive lowering of LDL cholesterol (LDL-C) levels to <100 mg/dL compared with a moderate reduction to 132 to 136 mg/dL decreased the progression of atherosclerosis in grafts. Low-dose anticoagulation did not significantly affect progression. Methods and Results-Approximately 3 years after the last trial visit, Clinical Center Coordinators contacted each patient by telephone to ascertain the occurrence of cardiovascular events and procedures. The National Death Index was used to ascertain vital status for patients who could not be contacted, Vital status was established for all but 3 of 1351 patients. Information on nonfatal events was available for 95% of surviving patients. A 30% reduction in revascularization procedures and 24% reduction in a composite clinical end point were observed in patients assigned to aggressive strategy compared with patients assigned to moderate strategy during 7.5 years of follow-up, P=0.0006 and 0.001, respectively. Reductions of 35% in deaths and 31% in deaths or myocardial infarctions with low-dose anticoagulation compared with placebo were also observed, P=0.008 and 0.003, respectively. Conclusions-The long-term clinical benefit observed during extended follow-up in patients assigned to the aggressive strategy is consistent with the angiographic findings of delayed atherosclerosis progression in grafts observed during the trial. The apparent long-term benefit of low-dose warfarin remains unexplained. C1 Maryland Med Res Inst, Post CABG Coordinating Ctr, Baltimore, MD 21210 USA. NHLBI, Clin Trials Grp, Bethesda, MD 20892 USA. NHLBI, Clin Trials Grp, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, Bethesda, MD 20892 USA. Montreal Heart Inst, Montreal, PQ H1T 1C8, Canada. Univ Minnesota, Heart Dis Prevent Clin, Minneapolis, MN USA. Univ Minnesota, Angiogram Reading Ctr, Minneapolis, MN USA. Minneapolis Heart Inst, Minneapolis, MN USA. Cedars Sinai Med Ctr, Los Angeles, CA 90048 USA. Baylor Coll Med, Houston, TX 77030 USA. Cleveland Clin Fdn, Cleveland, OH 44195 USA. RP Knatterud, GL (reprint author), Maryland Med Res Inst, Post CABG Coordinating Ctr, 600 Wyndhurst Ave, Baltimore, MD 21210 USA. FU NHLBI NIH HHS [N01-HC-75071, N01-HC-75072, N01-HC-75073] NR 19 TC 169 Z9 173 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 11 PY 2000 VL 102 IS 2 BP 157 EP + PG 10 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 332LA UT WOS:000088074700012 PM 10889125 ER PT J AU Brown, AL Mann, NC Daya, M Goldberg, MR Meischke, H Taylor, J Smith, K Osganian, S Cooper, L AF Brown, AL Mann, NC Daya, M Goldberg, MR Meischke, H Taylor, J Smith, K Osganian, S Cooper, L CA Rapid Early Action Coronary Treatm TI Demographic, belief, and situational factors influencing the decision to utilize emergency medical services among chest pain patients SO CIRCULATION LA English DT Article DE coronary disease; epidemiology; public policy ID ACUTE MYOCARDIAL-INFARCTION; RAPID-EARLY-ACTION; CORONARY-TREATMENT; CALL 911; DELAY; SYMPTOMS; CAMPAIGN; DESIGN; ONSET AB Background-Empirical evidence suggests that people value emergency medical services (EMS) but that they may not use the service when experiencing chest pain. This study evaluates this phenomenon and the factors associated with the failure to use EMS during a potential cardiac event. Methods and Results-Baseline data were gathered from a randomized, controlled community trial (REACT) that was conducted in 20 US communities. A random-digit-dial survey documented bystander intentions to use EMS for cardiac symptoms in each community. An emergency department surveillance system documented the mode of transport among chest pain patients in each community and collected ancillary data, including situational factors surrounding the chest pain event. Logistic regression identified factors associated with failure to use EMS. A total of 962 community members responded to the phone survey, and data were collected on 875 chest pain emergency department arrivals. The mean proportion of community members intending to use EMS during a witnessed cardiac event was 89%; the mean proportion of patients observed using the service was 23%, with significant geographic differences (range, 10% to 48% use). After controlling for covariates, non-EMS users were more likely to try antacids/aspirin and call a doctor and were less likely to subscribe to (or participate in) an EMS prepayment plan. Conclusion-The results of this study indicate that indecision, self-treatment, physician contact, and financial concerns may undermine a chest pain patient's intention to use EMS. C1 Univ Utah, Intermt Injury Control Res Ctr, Sch Med, Salt Lake City, UT 84108 USA. Oregon Hlth Sci Univ, Dept Emergency Med, Portland, OR 97201 USA. Univ Massachusetts, Sch Med, Dept Cardiol, Worcester, MA USA. Univ Washington, Dept Hlth Serv, Seattle, WA 98195 USA. Mississippi Univ Women, Div Hlth & Kinesiol, Columbus, MS USA. New England Res Inst, Watertown, MA 02172 USA. NHLBI, Bethesda, MD 20892 USA. RP Mann, NC (reprint author), Univ Utah, Intermt Injury Control Res Ctr, Sch Med, 410 Chipeta Way,Suite 222, Salt Lake City, UT 84108 USA. FU NHLBI NIH HHS [U01-HL-53141, U01-HL-53149, U01-HL-53412] NR 23 TC 76 Z9 78 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUL 11 PY 2000 VL 102 IS 2 BP 173 EP + PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 332LA UT WOS:000088074700014 PM 10889127 ER PT J AU Caplen, NJ Fleenor, J Fire, A Morgan, RA AF Caplen, NJ Fleenor, J Fire, A Morgan, RA TI dsRNA-mediated gene silencing in cultured Drosophila cells: a tissue culture model for the analysis of RNA interference SO GENE LA English DT Article DE post-transcriptional gene silencing; RNAi; S2 cells ID DOUBLE-STRANDED-RNA; CAENORHABDITIS-ELEGANS; C-ELEGANS; MESSENGER-RNA; DNA; EXPRESSION; NEUROSPORA; TRANSGENE; HELICASE; HOMOLOG AB RNA interference (RNAi) is a form of post-transcriptional gene silencing that has been described in a number of plant, nematode, protozoan, and invertebrate species. RNAi is characterized by a number of features: induction by double stranded RNA (dsRNA), a high degree of specificity, remarkable potency and spread across cell boundaries, and a sustained downregulation of the target gene. Previous studies of RNAi have examined this effect in whole organisms or in extracts thereof; we have now examined the induction of RNAi in tissue culture. A screen of mammalian cells from three different species showed no evidence for the specific down-regulation of gene expression by dsRNA. By contrast, RNAi was observed in Drosophila Schneider 2 (S2) cells. Green fluorescent protein (GFP) expression in S2 cells was inhibited in a dose-dependent manner by transfection of dsRNA corresponding to gfp when GFP was expressed either transiently or stably. This effect was structure- and sequence-specific in that: (1) little or no effect was seen when antisense (or sense) RNA was transfected; (2) an unrelated dsRNA did not reduce GFP expression, and (3) dsRNA corresponding to gfp had no effect on the expression of an unrelated target transgene. This invertebrate tissue culture model should allow facile assays for loss of function in a well-defined cellular system and facilitate further understanding of the mechanism of RNAi and the genes involved in this process. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Natl Human Genome Res Inst, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA. Carnegie Inst Washington, Dept Embryol, Baltimore, MD 21210 USA. RP Morgan, RA (reprint author), Natl Human Genome Res Inst, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA. RI Caplen, Natasha/H-2768-2016 OI Caplen, Natasha/0000-0002-0001-9460 NR 36 TC 159 Z9 184 U1 1 U2 15 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD JUL 11 PY 2000 VL 252 IS 1-2 BP 95 EP 105 DI 10.1016/S0378-1119(00)00224-9 PG 11 WC Genetics & Heredity SC Genetics & Heredity GA 337CG UT WOS:000088339400009 PM 10903441 ER PT J AU Huang, K Johnson, KD Petcherski, AG Vandergon, T Mosser, EA Copeland, NG Jenkins, NA Kimble, J Bresnick, EH AF Huang, K Johnson, KD Petcherski, AG Vandergon, T Mosser, EA Copeland, NG Jenkins, NA Kimble, J Bresnick, EH TI A HECT domain ubiquitin ligase closely related to the mammalian protein WWP1 is essential for Caenorhabditis elegans embryogenesis SO GENE LA English DT Article DE gene family; morphogenesis; ubiquitination; WW domain ID MOUSE; TRANSCRIPTION; CLONING; E6-AP; IDENTIFICATION; ORGANIZATION; EXPRESSION; SEQUENCES; PATHWAY; TARGETS AB The highly conserved ubiquitin/proteasome pathway controls the degradation of many critical regulatory proteins. Proteins are posttranslationally conjugated to ubiquitin through a concerted set of reactions involving activating (E1), conjugating (E2), and ligase (E3) enzymes. Ubiquitination targets proteins for proteolysis via the proteasome and may regulate protein function independent of proteolysis. We describe the cloning and functional analysis of new members of the HECT domain family of E3 ubiquitin ligases. Murine Wwp1 encoded a broadly expressed protein containing a C2 domain, four WW domains, and a catalytic HECT domain. A Caenorhabditis elegans gene was cloned encoding a HECT domain protein (CeWWP1), which was highly homologous to murine and human WWP1. Disruption of CeWwp1 via RNA interference yielded an embryonic lethal phenotype, despite the presence of at least six additional C. elegans genes encoding HECT domain proteins. The embryonic lethality was characterized by grossly abnormal morphogenesis during late embryogenesis, despite normal proliferation early in embryogenesis. CeWWP1 must therefore have unique and nonredundant functions critical for embryogenesis. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Wisconsin, Sch Med, Dept Pharmacol, Mol & Cellular Pharmacol Program, Madison, WI 53706 USA. Univ Wisconsin, Howard Hughes Med Inst, Madison, WI 53706 USA. Univ Wisconsin, Dept Biochem, Madison, WI 53706 USA. Pepperdine Univ, Div Nat Sci, Malibu, CA 90263 USA. NCI, Mammalian Genet Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Bresnick, EH (reprint author), Univ Wisconsin, Sch Med, Dept Pharmacol, Mol & Cellular Pharmacol Program, 387 Med Sci Ctr,1300 Univ Ave, Madison, WI 53706 USA. FU NIDDK NIH HHS [DK55700] NR 33 TC 27 Z9 28 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 11 PY 2000 VL 252 IS 1-2 BP 137 EP 145 DI 10.1016/S0378-1119(00)00216-X PG 9 WC Genetics & Heredity SC Genetics & Heredity GA 337CG UT WOS:000088339400013 PM 10903445 ER PT J AU Horkay, F McKenna, GB Deschamps, P Geissler, E AF Horkay, F McKenna, GB Deschamps, P Geissler, E TI Neutron scattering properties of randomly cross-linked polyisoprene gels SO MACROMOLECULES LA English DT Article ID SWOLLEN; NETWORKS AB Measurements are reported of the osmotic swelling and elastic properties of cross-linked polyisoprene gels swollen in toluene, as a function of the polymer volume fraction phi. The swelling pressure and shear modulus results may be combined to yield the Flory-Huggins interaction parameter chi for the cross-linked state of this system. It is found that chi can be expressed as chi = chi(0) + chi(1)phi + chi(2)phi(2), where chi(0) = 0.427, chi(1) = 0.000, and chi(2) = 0.112. Small-angle neutron scattering (SANS) measurements are also reported for the same gels. The scattering intensity associated with the spontaneous thermodynamic concentration fluctuations in the gels is compared with that, computed from the osmotic data. The agreement between the observed neutron intensities and the calculated values is good. The SANS measurements reveal two additional components to the scattered intensity: one due to the local elastic constraints of the crosslinks and the other being caused by dense clusters that show no appreciable deformation when subjected to an external elastic strain field. C1 NICHD, Sect Tissue Biophys & Biomimet, Lab Integrat & Med Biophys, NIH, Bethesda, MD 20892 USA. Texas Tech Univ, Dept Chem Engn, Lubbock, TX 79409 USA. European Synchrotron Radiat Facil, F-38043 Grenoble, France. Univ Grenoble 1, Spectrometrie Phys Lab, CNRS UMR 5588, F-38402 St Martin Dheres, France. RP Horkay, F (reprint author), NICHD, Sect Tissue Biophys & Biomimet, Lab Integrat & Med Biophys, NIH, 13 South Dr, Bethesda, MD 20892 USA. RI McKenna, Gregory/O-1134-2013 OI McKenna, Gregory/0000-0002-5676-9930 NR 35 TC 43 Z9 43 U1 0 U2 10 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0024-9297 J9 MACROMOLECULES JI Macromolecules PD JUL 11 PY 2000 VL 33 IS 14 BP 5215 EP 5220 DI 10.1021/ma0003001 PG 6 WC Polymer Science SC Polymer Science GA 333UF UT WOS:000088148600033 ER PT J AU Sorlie, PD Nieto, FJ Adam, E Folsom, AR Shahar, E Massing, M AF Sorlie, PD Nieto, FJ Adam, E Folsom, AR Shahar, E Massing, M TI A prospective study of cytomegalovirus, herpes simplex virus 1, and coronary heart disease - The Atherosclerosis Risk in Communities (ARIC) Study SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID SOLID-PHASE RADIOIMMUNOASSAY; CHLAMYDIA-PNEUMONIAE; INFECTION; ANTIBODY; ARTERY; RESTENOSIS; COHORT; AGENTS; LINK AB Background: Conflicting evidence exists implicating infectious disease in the pathological processes leading to coronary heart disease (CHD). The objective of this article is to describe the relationship of previous infection with cytomegalovirus (CMV) and herpes simplex virus 1 to incident CHD in a population-based cohort study. Methods: Using a nested case-cohort design from the Atherosclerosis Risk in Communities Study, antibody levels to CMV and herpes simplex virus 1 were determined in serum samples that bad been frozen at the baseline examination in participants free of CHD. Determinations were made in those who developed incident CHD (n=221) during follow-up of up to 5 years from baseline and in a stratified random sample of all participants (n=515). Results: The population with the highest antibody levels of CMV (approximately the upper 20%) showed an increased relative risk (RR) of CHD of 1.76 (95% confidence interval, 1.00-3.11), adjusting for age, sex, and race. After adjustment for additional covariates of hypertension, diabetes, years of education, cigarette smoking, low-density lipoprotein and high-density lipoprotein cholesterol levels, and fibrinogen level, the RR increased slightly. Based on a priori hypotheses, the RR of CHD at the highest antibody levels in individuals with diabetes was particularly large but with wide confidence intervals (RR, 9.2; 95% confidence interval, 1.8-47.0), and the interaction between high levels of antibody to CMV and diabetes was statistically significant (P=.05). There was no association of CHD with the highest herpes simplex virus 1 antibody levels (adjusted RR, 0.77; 95% confidence interval, 0.36-1.62). Conclusions: High levels of CMV antibodies are significantly associated with incident CHD. Infection with CMV, particularly in more susceptible disease states such as diabetes, may be an important risk factor for CHD. C1 NHLBI, NIH, Bethesda, MD 20892 USA. Johns Hopkins Sch Hyg & Publ Hlth, Dept Epidemiol, Baltimore, MD USA. Baylor Coll Med, Div Mol Virol, Houston, TX 77030 USA. Univ Minnesota, Div Epidemiol, Minneapolis, MN USA. Univ N Carolina, Dept Epidemiol, Chapel Hill, NC USA. RP Sorlie, PD (reprint author), NHLBI, NIH, 6701 Rockledge Dr,Mail Stop Code 7934, Bethesda, MD 20892 USA. FU NHLBI NIH HHS [N01-HC-55015, N01-HC-55016, N01-HC-55018] NR 23 TC 99 Z9 105 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 10 PY 2000 VL 160 IS 13 BP 2027 EP 2032 DI 10.1001/archinte.160.13.2027 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 330XZ UT WOS:000087988000014 PM 10888976 ER PT J AU Zhao, HC Jin, SQ Antinore, MJ Lung, FDT Fan, FY Blanck, P Roller, P Fornace, AJ Zhan, QM AF Zhao, HC Jin, SQ Antinore, MJ Lung, FDT Fan, FY Blanck, P Roller, P Fornace, AJ Zhan, QM TI The central region of Gadd45 is required for its interaction with p21/WAF1 SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE GADD45; p21; p53; growth suppression; cell cycle checkpoint ID CYCLIN-DEPENDENT KINASES; P53-REGULATED PROTEIN GADD45; P53-MEDIATED G(1) ARREST; TUMOR-SUPPRESSOR P53; DNA-REPLICATION; CDK INHIBITOR; WILD-TYPE; P21; PCNA; GENE AB Cell cycle arrest represents an important response to genotoxic stress and the tumor suppressor p53 has been described to act as a critical effector in this biological event. Upon stress, p53 becomes transcriptionally active and up-regulates the transcription of downstream effector genes, which contain p53 recognition sites in their regulatory regions. Among the genes activated are p21 and GADD45, each of which independently exhibits growth-suppressive activity. The Gadd45 protein has been described to form a complex with p21, and thus, work was undertaken to map the regions of Gadd45 involved in this interaction and to examine the roles of those two proteins in growth suppression. In this report, a Gadd45 overlapping peptide library and a series of Gadd45 deletion mutants were used to define the domains of Gadd45 involved in the association with p21. Results using both in vitro and in vivo methods have shown that the interaction of Gadd45 with p21 involves a central region of Gadd45. Interestingly, the pal-binding domain of Gadd45 also encodes the Cdc2-binding activity, indicating that the central region of Gadd45 may serve as an important "core," through which Gadd45 protein is able to present cross-talk with other cell cycle regulators. In addition, GADD45 inhibition of Cdc2 kinase activity was compared with Myd118 and CR6, two other members of the GADD45 family. GADD45 was shown to generate the strongest inhibitory effect on Cdc2 activity. Finally, results from short-term survival assays further demonstrated that p21 and GADD45 act upon different cellular pathways to exert their growth-suppressive function. (C) 2000 Academic Press. C1 Univ Pittsburgh, Sch Med, Pittsburgh Canc Inst, Dept Radiat Oncol, Pittsburgh, PA 15213 USA. NCI, Gene Response Sect, NIH, Bethesda, MD 20892 USA. NCI, Med Chem Lab, DBS, NIH, Bethesda, MD 20892 USA. RP Zhan, QM (reprint author), Univ Pittsburgh, Sch Med, Pittsburgh Canc Inst, Dept Radiat Oncol, Pittsburgh, PA 15213 USA. RI Fornace, Albert/A-7407-2008 OI Fornace, Albert/0000-0001-9695-085X NR 42 TC 45 Z9 49 U1 0 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD JUL 10 PY 2000 VL 258 IS 1 BP 92 EP 100 DI 10.1006/excr.2000.4906 PG 9 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 333XN UT WOS:000088156200010 PM 10912791 ER PT J AU Spector, M Nguyen, VA Sheng, XN He, LS Woodward, J Fan, SJ Baumgarten, CM Kunos, G Dent, P Gao, B AF Spector, M Nguyen, VA Sheng, XN He, LS Woodward, J Fan, SJ Baumgarten, CM Kunos, G Dent, P Gao, B TI Activation of mitogen-activated protein kinases is required for alpha(1)-adrenergic agonist-induced cell scattering in transfected HepG2 cells SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE cell hypertrophy; protein kinase C; AP1 ID HEPATOCYTE GROWTH-FACTOR; CARDIAC MYOCYTE HYPERTROPHY; GENE-EXPRESSION; PHOSPHATIDYLINOSITOL 3-KINASE; ADRENERGIC-RECEPTORS; SIGNALING PATHWAY; RAT HEPATOCYTES; SMOOTH-MUSCLE; MAP KINASE; BETA-2-ADRENERGIC RECEPTORS AB Activation of alpha(1B)-adrenergic receptors (alpha(1B)AR) by phenylephrine (PE) induces scattering of HepG2 cells stably transfected with the alpha(1B)AR (TFG2 cells). Scattering was also observed after stimulation of TFG2 cells with phorbol myristate acetate (PMA) but not with hepatocyte growth factor/scatter factor, epidermal growth factor, or insulin. PMA but not phenylephrine rapidly activated PKC alpha in TFG2 cells, and the highly selective PKC inhibitor bisindolylmaleimide (GFX) completely abolished PMA-induced but not PE-induced scattering PE rapidly activated p44/42 mitogen-activated protein kinase (MAPK), p38 MAPK, c-Jun N-terminal kinase (JNK), and AP1 (c-fos/c-jun), Selective blockade of p42/44 MAPK activity by PD98059 or by transfection of a MEK1 dominant negative adenovirus significantly inhibited the PE-induced scattering of TFG2 cells. Selective inhibition of p38 MAPK by SB203850 or SB202190 also blocked PE-induced scattering, whereas treatment of TFG2 cells with the PI3 kinase inhibitors LY294002 or wortmannin did not inhibit PE-induced scattering. Blocking JNK activation with a dominant negative mutant of JNK or blocking AP1 activation with a dominant negative mutant of c-jun (TAM67) significantly inhibited PE-induced cell scattering These data indicate that PE-induced scattering of TFG2 cells is mediated by complex mechanisms, including activation of p42/44 MAPK, p38 MAPK, and JNK, Cell spreading has been reported to play important roles in wound repair, tumor invasion, and metastasis, Therefore, catecholamines acting via the alpha(1)AR may modulate these physiological and pathological processes. (C) 2000 Academic Press. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Physiol, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Radiat Oncol, Richmond, VA 23298 USA. NCI, NIH, Bethesda, MD 20892 USA. Albert Einstein Coll Med, Long Isl Jewish Med Ctr, Dept Radiat Oncol, New York, NY USA. RP Gao, B (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Box 980613, Richmond, VA 23298 USA. FU NCI NIH HHS [R29CA72681]; NHLBI NIH HHS [R01 HL046764-08]; NIAAA NIH HHS [R03AA11823, R01AA12637] NR 53 TC 16 Z9 18 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD JUL 10 PY 2000 VL 258 IS 1 BP 109 EP 120 DI 10.1006/excr.2000.4907 PG 12 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 333XN UT WOS:000088156200012 PM 10912793 ER PT J AU Aharonovitz, O Zaun, HC Balla, T York, JD Orlowski, J Grinstein, S AF Aharonovitz, O Zaun, HC Balla, T York, JD Orlowski, J Grinstein, S TI Intracellular pH regulation by Na+/H+ exchange requires phosphatidylinositol 4,5-bisphosphate SO JOURNAL OF CELL BIOLOGY LA English DT Article DE amiloride; ATP depletion; NA(+)/H+ antiport; phosphoinositide ID PLECKSTRIN-HOMOLOGY DOMAINS; CAMP-MEDIATED INHIBITION; CELL-VOLUME REGULATION; ATP DEPENDENCE; POTASSIUM CHANNELS; PLASMA-MEMBRANE; ISOFORM-1 NHE1; PHOSPHORYLATION; IDENTIFICATION; SENSITIVITY AB The carrier-mediated, electroneutral exchange of Na+ for H+ across the plasma membrane does not directly consume metabolic energy. Nevertheless, acute depletion of cellular ATP markedly decreases transport. We analyzed the possible involvement of polyphosphoinositides in the metabolic regulation of NHE1, the ubiquitous isoform of the Na+/H+ exchanger. Depletion of ATP was accompanied by a marked reduction of plasmalemmal phosphatidylinositol 4,5-bisphosphate (PIP2) content. Moreover, sequestration or hydrolysis of plasmalemmal PIP2, in the absence of ATP depletion, was associated with profound inhibition of NHE1 activity. Examination of the primary structure of the COOH-terminal domain of NHE1 revealed two potential PIP2-binding motifs. Fusion proteins encoding these motifs bound PIP2 in vitro. When transfected into antiport-deficient cells, mutant forms of NHE1 lacking the putative PIP2-binding domains had greatly reduced transport capability, implying that association with PIP2 is required for optimal activity. These findings suggest that NHE1 activity is modulated by phosphoinositides and that the inhibitory effect of ATP depletion may be attributable, at least in part, to the accompanying net dephosphorylation of PIP2. C1 Hosp Sick Children, Div Cell Biol, Res Inst, Cell Biol Programme, Toronto, ON M5G 1X8, Canada. McGill Univ, Dept Physiol, Montreal, PQ H3G 1Y6, Canada. NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Duke Univ, Med Ctr, Dept Pharmacol & Canc Biol, Durham, NC 27710 USA. RP Grinstein, S (reprint author), Hosp Sick Children, Div Cell Biol, Res Inst, Cell Biol Programme, 555 Univ Ave, Toronto, ON M5G 1X8, Canada. OI Balla, Tamas/0000-0002-9077-3335 FU NHLBI NIH HHS [R01-HL 55672, R01 HL055672] NR 47 TC 139 Z9 142 U1 1 U2 3 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUL 10 PY 2000 VL 150 IS 1 BP 213 EP 224 DI 10.1083/jcb.150.1.213 PG 12 WC Cell Biology SC Cell Biology GA 334UC UT WOS:000088203200022 PM 10893269 ER PT J AU Hook, S Prout, M Camberis, M Konig, M Zimmer, A Van Heeke, G Le Gros, G AF Hook, S Prout, M Camberis, M Konig, M Zimmer, A Van Heeke, G Le Gros, G TI Th2-dependent airway eosinophilia is regulated by preproenkephalin SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE preproenkephalin; Th2; eosinophils; cytokine ID MESSENGER-RNA; PROENKEPHALIN-A; OPIOID RECEPTOR; LYMPHOCYTES-T; IN-VITRO; CELLS; ENKEPHALIN; EXPRESSION; RESPONSES; PEPTIDES AB We have used preproenkephalin (PPNK)-deficient mice to study the role of PPNK in the development of airway inflammation. Airway eosinophilia was established by either sensitization followed by airway challenge with OVA or by infection with Nippostrongylus brasiliensis. Both models induce a strong Th2 immune response, characterized by an IL-5-dependent airway eosinophilia. We observed that although the accumulation of lymphocytes in the airways of PPNK-deficient mice was similar to that induced in control mice, IL-5 production and eosinophil infiltration were reduced. We conclude from this work that PPNK has a role in enhancing Th2 cell function and that this molecule may be an important target in asthma immunotherapy. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Wellington Sch Med, Malaghan Inst Med Res, Wellington, New Zealand. NIMH, Genet Lab, Unit Mouse Genet, Bethesda, MD 20892 USA. Novartis Horsham Res Ctr, Horsham, W Sussex, England. RP Hook, S (reprint author), Wellington Sch Med, Malaghan Inst Med Res, POB 7060, Wellington, New Zealand. RI Le Gros, Graham/C-6725-2011; Zimmer, Andreas/B-8357-2009 NR 21 TC 14 Z9 15 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 JUL 10 PY 2000 VL 107 IS 1 BP 59 EP 65 DI 10.1016/S0165-5728(00)00243-5 PG 7 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 318AG UT WOS:000087260700007 PM 10808051 ER PT J AU Hatano, H Vogel, S Yoder, C Metcalf, JA Dewar, R Davey, RT Polis, MA AF Hatano, H Vogel, S Yoder, C Metcalf, JA Dewar, R Davey, RT Polis, MA TI Pre-HAART HIV burden approximates post-HAART viral levels following interruption of therapy in patients with sustained viral suppression SO AIDS LA English DT Article DE HIV; highly active antiretroviral therapy; viral load; viral load setpoint ID ACTIVE ANTIRETROVIRAL THERAPY; RNA MEASUREMENTS; PLASMA; INFECTION; CELLS; LOAD; DISCONTINUATION; INTERLEUKIN-2; PERSISTENCE; REPLICATION AB Objective: To evaluate the relationship between the HIV viral burden in individuals prior to receiving highly-active antiretroviral therapy (HAART) and the viral burden after withdrawal of HAART. Design and setting: Retrospective cohort study at the National Institutes of Health, Bethesda, Maryland, USA. Patients: Fourteen HIV-infected patients who achieved and maintained viral control on HAART who subsequently discontinued HAART. Main outcome measures: Pre- and post-HAART viral loads measured from plasma or serum. Results: Patients achieved viral control (< 500 copies/ml) on HAART in a median 28 days (range, 15-490 days; mean, 72 days), maintained viral control for a median 661 days (range, 53-1067 days; mean, 611 days), and subsequently discontinued HAART for a median 49 days (range, 14-196 days; mean, 73 days). The median difference between the pre- and post-HAART viral loads was 0.16 log(10) (range, -0.72 to 1.05 log(10); mean, 0.19 log(10)). The median absolute difference between the pre- and post HAART viral loads was 0.43 log(10) (range, 0.06-1.05 log(10); mean, 0.46 log(10)). Nine individuals had post-HAART values higher than pre-HAART values, five had lower values. Median duration between pre- and post-HAART viral load measurements was 1757 days (range, 117-3177 days; mean, 1756 days), or 4.8 years. Conclusions: After discontinuing HAART, individuals had rebounds in their viral burdens approximating pre-HAART levels, even after a significant lapse of time approaching 5 years. Our data suggest that an intrinsic viral load set-point may exist, and that a single interruption of an effective regimen with viral suppression for almost 2 years does not significantly alter this set-point. (C) 2000 Lippincott Williams & Wilkins. C1 NIAID, Immunoregulat Lab, Bethesda, MD 20892 USA. NIH, Dept Crit Care Med, Warren Magnuson Clin Ctr, Bethesda, MD 20892 USA. Sci Applicat Int Corp, Frederick, MD USA. RP Polis, MA (reprint author), NIAID, Immunoregulat Lab, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Polis, Michael/0000-0002-9151-2268 FU NCI NIH HHS [N01-CO-56000] NR 33 TC 72 Z9 73 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 7 PY 2000 VL 14 IS 10 BP 1357 EP 1363 DI 10.1097/00002030-200007070-00008 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 333RW UT WOS:000088145400008 PM 10930150 ER PT J AU Lambert, JS Watts, DH Mofenson, L Stiehm, ER Harris, DR Bethel, J Whitehouse, J Jimenez, E Gandia, J Scott, G O'Sullivan, MJ Kovacs, A Stek, A Shearer, WT Hammill, H van Dyke, R Maupin, R Silio, M Fowler, MG AF Lambert, JS Watts, DH Mofenson, L Stiehm, ER Harris, DR Bethel, J Whitehouse, J Jimenez, E Gandia, J Scott, G O'Sullivan, MJ Kovacs, A Stek, A Shearer, WT Hammill, H van Dyke, R Maupin, R Silio, M Fowler, MG CA Pediat AIDS Clin Trials Grp 185 Te TI Risk factors for preterm birth, low birth weight, and intrauterine growth retardation in infants born to HIV-infected pregnant women receiving zidovudine SO AIDS LA English DT Article DE HIV; vertical infection; pregnancy; preterm birth; low birth weight; intrauterine growth retardation; antiretroviral therapy; zidovudine ID HUMAN-IMMUNODEFICIENCY-VIRUS; MOTHER-TO-CHILD; CLINICAL-TRIALS GROUP; PERINATAL TRANSMISSION; ASSOCIATION; OUTCOMES; TYPE-1; AIDS; PREMATURITY; ALCOHOL AB Objective: To evaluate independent contributions of maternal factors to adverse pregnancy outcomes (APO) in HIV-infected women receiving antiretroviral therapy (ART). Design: Risk factors for preterm birth (< 37 weeks gestation), low birth weight (LBW) (< 2500 g), and intrauterine growth retardation (IUGR) (birth weight < 10(th) percentile for gestational age) examined in 497 HIV-infected pregnant women enrolled in PACTG 185, a perinatal clinical trial. Methods: HIV RNA copy number, culture titer, and CD4 lymphocyte counts were measured during pregnancy. Information collected included antenatal use of cigarettes, alcohol, illicit drugs; ART; obstetric history and complications. Results: Eighty-six percent were minority race/ethnicity; 86% received antenatal monotherapy, predominantly zidovudine (ZDV), and 14% received combination antiretrovirals. Preterm birth occurred in 17%, LBW in 13%, IUGR in 6%. Risk of preterm birth was independently associated with prior preterm birth [odds ratio (OR) 3.34; P < 0.001], multiple gestation (OR, 6.02; P = 0.011), antenatal alcohol use (OR, 1.91; P = 0.038), and antenatal diagnosis of genital herpes (OR, 0.24; P = 0.022) or pre-eclampsia (OR, 6.36; P = 0.025). LEW was associated with antenatal diagnosis of genital herpes (OR, 0.08; P = 0.014) and pre-eclampsia (OR, 5.25; P = 0.049), and baseline HIV culture titer (OR, 1.41; P = 0.037). IUGR was associated with multiple gestation (OR, 8.20; P = 0.010), antenatal cigarette use (OR, 3.60; P = 0.008), and pre-eclampsia (OR, 12.90; P = 0.007). Maternal immune status and HIV RNA copy number were not associated with APO. Conclusions: Risk factors for APO in antiretroviral treated HIV-infected women are similar to those reported for uninfected women. These data suggest that provision of prenatal care and ART may reduce APO. (C) 2000 Lippincott Williams & Wilkins. C1 Univ Maryland, Inst Human Virol, Baltimore, MD 21201 USA. Case Western Reserve Univ Hosp, Cleveland, OH 44106 USA. Univ Med & Dent New Jersey, Newark, NJ 07103 USA. Childrens Hosp, Boston, MA 02115 USA. Brigham & Womens Hosp, Boston, MA 02115 USA. Boston Univ, Med Ctr, Boston, MA USA. Univ Calif Los Angeles, Med Ctr, Los Angeles, CA 90024 USA. Johns Hopkins Univ, Sch Med, Baltimore, MD USA. Univ Maryland, Baltimore, MD 21201 USA. Baylor Coll Med, Houston, TX USA. Univ Texas, Sch Med, Houston, TX USA. Columbia Presbyterian Med Ctr, New York, NY 10032 USA. Univ Miami, Sch Med, Miami, FL USA. Mt Sinai Sch Med, New York, NY USA. NYU, Med Ctr, New York, NY 10016 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. San Francisco Gen Hosp, San Francisco, CA 94110 USA. Univ Calif San Diego, La Jolla, CA 92093 USA. Univ N Carolina, Chapel Hill, NC USA. Univ Illinois, Chicago, IL USA. City Hosp, San Juan, PR USA. Ramon Ruiz Arnau Univ Hosp, Bayamon, PR USA. SUNY Stony Brook, Stony Brook, NY 11794 USA. Childrens Hosp, Detroit, MI 48201 USA. Hutzel Hosp, Detroit, MI 48201 USA. Albany Med Ctr, Albany, NY USA. Univ Texas, SW Med Ctr, Dallas, TX USA. Howard Univ Hosp, Washington, DC USA. Univ So Calif, Los Angeles Cty Med Ctr, Los Angeles, CA 90033 USA. Univ Florida, Hlth Sci Ctr, Jacksonville, FL 32209 USA. Univ Colorado, Hlth Sci Ctr, Denver, CO USA. Virginia Commonwealth Univ, Richmond, VA USA. Methodist Hosp, Reg Med Ctr Memphis, St Jude Childrens Res Hosp, Memphis, TN USA. Univ Puerto Rico, Sch Med, San Juan, PR 00936 USA. Childrens Hosp Philadelphia, Philadelphia, PA 19104 USA. Univ Penn, Philadelphia, PA 19104 USA. Thomas Jefferson Univ Hosp, Philadelphia, PA 19107 USA. St Christophers Hosp Children, Philadelphia, PA 19133 USA. Temple Univ, Philadelphia, PA 19122 USA. Childrens Hosp, Seattle, WA USA. Med Ctr, Seattle, WA USA. Bronx Lebanon Hosp, Bronx, NY USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. Washington Hosp Ctr, Washington, DC 20010 USA. Childrens Hosp Kings Daughter & Sentara Norfolk G, Norfolk, VA USA. Tulane Univ, New Orleans, LA 70118 USA. Louisiana State Univ, New Orleans, LA USA. Med Ctr Cent Massachusetts, Worcester, MA USA. Baystate Med Ctr, Springfield, MA USA. Univ Connecticut, Ctr Hlth, Farmington, CT USA. Childrens Med Ctr, Farmington, CT USA. Univ Alabama, Birmingham, AL USA. SUNY Hlth Sci Ctr, Brooklyn, NY 11203 USA. George Washington Univ, Med Ctr, Washington, DC 20037 USA. Univ Minnesota, Minneapolis, MN USA. Harvard Univ, Sch Publ Hlth, PACTG Stat & Data Anal Ctr, Boston, MA 02115 USA. NHLBI, NIH, Bethesda, MD 20892 USA. NICHHD, NIH, Bethesda, MD 20892 USA. NIAID, NIH, Bethesda, MD 20892 USA. Westat, Rockville, MD USA. NABI, Boca Raton, FL USA. Glaxo Wellcome, Res Triangle Pk, NC USA. Quest Diagnost, Baltimore, MD USA. RP Lambert, JS (reprint author), Univ Maryland, Inst Human Virol, 725 W Lombard St, Baltimore, MD 21201 USA. OI Mofenson, Lynne/0000-0002-2818-9808 FU NHLBI NIH HHS [HL-57128]; NIAID NIH HHS [AI-27565]; NICHD NIH HHS [HD-33162] NR 39 TC 50 Z9 53 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 7 PY 2000 VL 14 IS 10 BP 1389 EP 1399 DI 10.1097/00002030-200007070-00012 PG 11 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 333RW UT WOS:000088145400012 PM 10930154 ER PT J AU Dunn, DT Simonds, RJ Bulterys, M Kalish, LA Moye, J de Maria, A Kind, C Rudin, C Denamur, E Krivine, A Loveday, C Newell, ML AF Dunn, DT Simonds, RJ Bulterys, M Kalish, LA Moye, J de Maria, A Kind, C Rudin, C Denamur, E Krivine, A Loveday, C Newell, ML TI Interventions to prevent vertical transmission of HIV-1: effect on viral detection rate in early infant samples SO AIDS LA English DT Article DE vertical transmission; viral detection rate; timing of transmission; interventions ID POLYMERASE CHAIN-REACTION; IMMUNODEFICIENCY-VIRUS TYPE-1; NEONATAL-PERIOD; ZIDOVUDINE PROPHYLAXIS; DISEASE PROGRESSION; INFECTION; SENSITIVITY; INTRAUTERINE; DIAGNOSIS; ASSAY AB Objective: To determine whether mode of delivery or the use of maternal or neonatal antiretroviral prophylaxis influence the age when HIV-1 can first be detected in infected infants, particularly the probability of detection at birth. Methods: In a collaboration between four multicentre studies, data on 422 HIV-1 infected infants who were assessed by HIV-1 DNA PCR or cell culture before 14 days of age were analysed. Weibull mixture models were used to estimate the cumulative proportion of infants with detectable levels of HIV-1 according to use of maternal/neonatal antiretroviral therapy (mainly zidovudine monotherapy) and mode of delivery. Results: HIV-1 was detected in 162 infants (38%) when they were first tested, at a median age of 2 days. At birth, it was estimated that 36% [95% confidence interval (CI), 31-41%] of infants have levels of virus that can be detected by DNA PCR or cell culture. This percentage was not associated with either mode of delivery (35% for vaginal delivery versus 40% for cesarean section delivery; P = 0.4) or the use of maternal or neonatal antiretroviral prophylaxis. Among infants with undetectable levels of HIV-1 at birth, the median time to viral detectability was estimated to be 14.8 days (95% CI, 12.9-16.8 days). This time was increased by 15% (95% CI, -11 to 48%; P = 0.3) among infants who were exposed to antiretroviral therapy postnatally compared with infants who were not exposed. No effect was observed for mode of delivery. Conclusions: The outcome of an early virological test for HIV-1 is thought to be related directly to the timing of transmission and cesarean section delivery primarily reduces the risk of intrapartum transmission. The absence of an association between mode of delivery and viral detectability at birth was therefore unexpected. There was no evidence that foetal or neonatal exposure to prophylactic zidovudine delays substantially the diagnosis of infection, although this cannot be inferred for combination antiretroviral therapy. (C) 2000 Lippincott Williams & Wilkins. C1 UCL, Inst Child Hlth, Dept Epidemiol & Publ Hlth, London, England. Ctr Dis Control & Prevent, Atlanta, GA USA. New England Res Inst, Watertown, MA 02172 USA. NICHHD, NIH, Bethesda, MD 20892 USA. Univ Genoa, Dept Internal Med, I-16126 Genoa, Italy. Kantonsspital, Div Neonatol, St Gallen, Switzerland. Univ Basel, Childrens Hosp, Basel, Switzerland. Hop Robert Debre, Biochim Genet Lab, F-75019 Paris, France. Hop St Vincent de Paul, Serv Bacteriol, F-75674 Paris, France. Royal Free & Univ Coll Med Sch, Dept Retrovirol, London, England. RP Dunn, DT (reprint author), MRC, Clin Trials Unit, 222 Euston Rd, London NW1 2DA, England. RI SHCS, MoCHIV/G-4081-2011; SHCS, all/G-4072-2011; SHCS, ch/G-4077-2011; SHCS, int. coll. B/G-4090-2011; de maria, andrea/F-7116-2016; OI de maria, andrea/0000-0001-5782-333X; moye, john/0000-0001-9976-8586; Newell, Marie-Louise/0000-0002-1074-7699 NR 31 TC 27 Z9 28 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD JUL 7 PY 2000 VL 14 IS 10 BP 1421 EP 1428 DI 10.1097/00002030-200007070-00016 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 333RW UT WOS:000088145400016 PM 10930158 ER PT J AU Lovett-Racke, AE Smith, ME Arredondo, LR Bittner, PS Ratts, RB Shive, CL Forsthuber, TG Racke, MK AF Lovett-Racke, AE Smith, ME Arredondo, LR Bittner, PS Ratts, RB Shive, CL Forsthuber, TG Racke, MK TI Developmentally regulated gene expression of Th2 cytokines in the brain SO BRAIN RESEARCH LA English DT Article DE experimental allergic encephalomyelitis; GATA-3; immune-privilege; interleukin-4; interleukin-10 ID EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; MYELIN BASIC-PROTEIN; NERVE GROWTH-FACTOR; MULTIPLE-SCLEROSIS; IMMUNE PRIVILEGE; MESSENGER-RNA; MICE; IL-4; INTERLEUKIN-4 AB Given the critical role of cytokines in the regulation of an inflammatory response, we investigated whether certain cytokines are expressed in the brains of normal mice during maturation that could contribute to the immune-privileged nature of the CNS or potentially influence an immune-mediated illness such as experimental allergic encephalomyelitis. The gene expression of IFN gamma (Th1 cytokine) and IL-4 (Th2 cytokine) was analyzed in the brain of several strains of mice. IFN gamma was not detectable. However, IL-4 was present in the brains of neonatal mice, but not adult mice. Resident CNS cells are believed to be the source of the IL-4, because mice deficient in T cells (SCID and RAG2-/-) expressed the IL-4 gene in the CNS. Further analysis indicated that the gene expression of the Th2 cytokine transcription factor, GATA-3, correlated with IL-4 and IL-10 expression in the brain. Since GATA-3-deficient mice have an abnormal CNS, brain-derived Th2 cytokines may play an important role in CNS development, as well as potentially contribute to the immune-privileged nature of the brain. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Texas, SW Med Ctr, Dept Neurol, Dallas, TX 75235 USA. Washington Univ, Sch Med, Dept Neurol, St Louis, MO 63110 USA. NIAID, DAIDS, NIH, Bethesda, MD 20892 USA. Case Western Reserve Univ, Dept Pathol, Cleveland, OH 44106 USA. Univ Texas, SW Med Ctr, Ctr Immunol, Dallas, TX USA. RP Lovett-Racke, AE (reprint author), Univ Texas, SW Med Ctr, Dept Neurol, 5323 Harry Hines Blvd, Dallas, TX 75235 USA. RI Lovett-Racke, Amy/E-3545-2011 NR 37 TC 20 Z9 20 U1 0 U2 0 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 7 PY 2000 VL 870 IS 1-2 BP 27 EP 35 DI 10.1016/S0006-8993(00)02398-2 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 330FE UT WOS:000087951500004 PM 10869498 ER PT J AU Morcock, DR Sowder, RC Casas-Finet, JR AF Morcock, DR Sowder, RC Casas-Finet, JR TI Role of the histidine residues of visna virus nucleocapsid protein in metal ion and DNA binding SO FEBS LETTERS LA English DT Article DE histidine residues; visna virus nucleocapsid protein; metal ion binding; DNA binding; zinc finger domains ID HUMAN-IMMUNODEFICIENCY-VIRUS; DETECTED MAGNETIC-RESONANCE; ZINC FINGER PEPTIDES; LEUKEMIA-VIRUS; ACID; SEQUENCE; TYPE-1; PHOSPHORESCENCE; RETROVIRUSES; FLUORESCENCE AB Zinc finger (ZF) domains in retroviral nucleocapsid proteins usually contain one histidine per metal ion coordination complex (Cys-X-2-Cys-X-4-His-X-4-Cys). Visna virus nucleocapsid protein, p8, has two additional histidines (in the second of its two ZFs) that could potentially bind metal ions. Absorption spectra of cobalt-bound ZF2 peptides were altered by Cys alkylation and mutation, but not by mutation of the extra histidines. Our results show that visna p8 ZFs involve three Cys and one His in the canonical spacing in metal ion coordination, and that the two additional histidines appear to interact with nucleic acid bases in p8-DNA complexes. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 NCI, AIDS Vaccine Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Casas-Finet, JR (reprint author), NCI, AIDS Vaccine Program, SAIC Frederick, Frederick Canc Res & Dev Ctr, Bldg 535-4,POB B, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 16 TC 9 Z9 9 U1 1 U2 6 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 7 PY 2000 VL 476 IS 3 BP 190 EP 193 DI 10.1016/S0014-5793(00)01723-3 PG 4 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 335LM UT WOS:000088245000016 PM 10913611 ER PT J AU Bezrukov, SM Kullman, L Winterhalter, M AF Bezrukov, SM Kullman, L Winterhalter, M TI Probing sugar translocation through maltoporin at the single channel level SO FEBS LETTERS LA English DT Article DE sugar transport; ion channel reconstitution; current fluctuation; noise analysis ID ESCHERICHIA-COLI; OUTER-MEMBRANE; SALMONELLA-TYPHIMURIUM; RATE CONSTANTS; TRANSPORT; NOISE; LAMB; CONDUCTANCE AB Sugar permeation through maltoporin of Escherichia coli, a trimer protein that facilitates maltodextrin translocation across outer bacterial membranes, was investigated at the single channel level. For large sugars, such as maltohexaose, elementary events of individual sugar molecule penetration into the channel a ere readily observed. At small sugar concentrations an elementary event consists of maltoporin channel closure by one third of its initial conductance in sugar-free solution. Statistical anal? sis of such closures at higher sugar concentrations shows that all three pores of the maltoporin channel transport sugars independently. interestingly, while channel conductance is only slightly asymmetric showing about 10% higher values at -200 mV than at +200 mV (from the side of protein addition), asymmetry in dependence of the sugar binding constant on the voltage polarity is about 20 times higher. Combining our data with observations made with bacteriophage-h we conclude that the sugar residence time is much more sensitive to (and is decreased by) voltages that are negative from the intra-cell side of the bacterial membrane. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 NICHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. Univ Basel, Biozentrum, Dept Biophys Chem, Basel, Switzerland. CNRS, IPBS, Toulouse, France. RP Bezrukov, SM (reprint author), NICHD, Lab Phys & Struct Biol, NIH, Bldg 9,Room 1E-122, Bethesda, MD 20892 USA. NR 16 TC 55 Z9 55 U1 0 U2 1 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 7 PY 2000 VL 476 IS 3 BP 224 EP 228 DI 10.1016/S0014-5793(00)01753-1 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 335LM UT WOS:000088245000023 PM 10913618 ER PT J AU Fan, YX McPhie, P Miles, EW AF Fan, YX McPhie, P Miles, EW TI Thermal repair of tryptophan synthase mutations in a regulatory intersubunit salt bridge - Evidence from arrhenius plots, absorption spectra, and primary kinetic isotope effects SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ULTRAVIOLET VISIBLE SPECTROSCOPY; ALPHA(2)BETA(2) COMPLEX REVEALS; INDUCED CONFORMATIONAL-CHANGES; SALMONELLA-TYPHIMURIUM; BIENZYME COMPLEX; BETA-SUBUNIT; ALPHA-SUBUNIT; ESCHERICHIA-COLI; 3-DIMENSIONAL STRUCTURE; ACTIVE-SITE AB This work is aimed at understanding how protein structure and conformation regulate activity and allosteric communication in the tryptophan synthase alpha(2)beta(2) complex from Salmonella typhimurium. Previous erystallographic and kinetic results suggest that both monovalent cations and a salt bridge between alpha subunit Asp(56) and beta subunit Lys(167) play allosteric roles. Here we show that mutation of either of these salt bridging residues produced deleterious effects that could be repaired by increased temperature in combination with CsCl or with NaCl plus an alpha subunit ligand, alpha-glycerol 3-phosphate. Arrhenius plots of the activity data under these conditions were nonlinear, The same conditions yielded temperature-dependent changes in the equilibrium distribution of enzyme-substrate intermediates and in primary kinetic isotope effects. We correlate the results with a model in which the mutant enzymes are converted by increased temperature from a low activity, "open" conformation to a high activity, "closed" conformation under certain conditions. The allosteric ligand and different monovalent cations affected the equilibrium between the open and closed forms. The results suggest that alpha subunit Asp(56) and beta subunit Lys(167) are not essential for catalysis and for allosteric communication between the alpha and beta subunits but that their mutual interaction is important in stabilization of the active, closed form of the alpha(2)beta(2) complex. C1 NIDDK, Sect Enzyme Struct & Funct, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Miles, EW (reprint author), NIDDK, Sect Enzyme Struct & Funct, Lab Biochem & Genet, NIH, Bldg 8,Rm 225,8 Ctr Dr MSC 0830, Bethesda, MD 20892 USA. NR 31 TC 6 Z9 6 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20302 EP 20307 DI 10.1074/jbc.M001135200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500014 PM 10801803 ER PT J AU Seo, MS Kang, SW Kim, K Baines, IC Lee, TH Rhee, SG AF Seo, MS Kang, SW Kim, K Baines, IC Lee, TH Rhee, SG TI Identification of a new type of mammalian peroxiredoxin that forms an intramolecular disulfide as a reaction intermediate SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ALKYL HYDROPEROXIDE REDUCTASE; THIOL-SPECIFIC ANTIOXIDANT; ACTIVATED PROTEIN-KINASE; NECROSIS-FACTOR-ALPHA; THIOREDOXIN PEROXIDASE; SIGNAL-TRANSDUCTION; CRYSTAL-STRUCTURE; OXIDATIVE STRESS; CELL-SURVIVAL; CLONING AB Peroxidases of the peroxiredoxin (Prx) family contain a Cys residue that is preceded by a conserved sequence in the NH2-terminal region. A new type of mammalian Prx, designated PrxV, has now been identified as the result of a data base search with this conserved Cys-containing sequence. The 162-amino acid PrxV shares only similar to 10% sequence identity with previously identified mammalian Prx enzymes and contains Cys residues at positions 73 and 152 in addition to that (Cys(48)) corresponding to the conserved Cys. Analysis of mutant human PrxV proteins in which each of these three Cys residues was individually replaced with serine suggested that the sulfhydryl group of Cys(48) is the site of oxidation by peroxides and that oxidized Cys(48) reacts with the sulfhydryl group of Cys(152) to form an intramolecular disulfide linkage. The oxidized intermediate of PrxV is thus distinct from those of other Prx enzymes, which form either an intermolecular disulfide or a sulfenic acid intermediate. The disulfide formed by PrxV is reduced by thioredoxin but not by glutaredoxin or glutathione. Thus, PrxV mutants lacking Cys(48) or Cys(152) showed no detectable thioredoxin-dependent peroxidase activity, whereas mutation of Cys(73) had no effect on activity. Immunoblot analysis revealed that PrxV is widely expressed in rat tissues and cultured mammalian cells and is localized intracellularly to cytosol, mitochondria, and peroxisomes. The peroxidase function of PrxV in vivo was demonstrated by the observations that transient expression of the wild-type protein, but not that of the Cys(48) mutant, in NIH 3T3 cells inhibited H2O2 accumulation and activation of c-Jun NH2-terminal kinase induced by tumor necrosis factor-alpha. C1 NHLBI, Lab Cell Signaling, NIH, Bethesda, MD 20892 USA. NHLBI, Dept Extramural Affairs, NIH, Bethesda, MD 20892 USA. Chonnam Natl Univ, Dept Food & Nutr, Kwangju 500757, South Korea. RP Rhee, SG (reprint author), NHLBI, Lab Cell Signaling, NIH, Bldg 3,Rm 120, Bethesda, MD 20892 USA. NR 38 TC 279 Z9 300 U1 1 U2 8 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20346 EP 20354 DI 10.1074/jbc.M001943200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500019 PM 10751410 ER PT J AU Tridandapani, S Lyden, TW Smith, JL Carter, JE Coggeshall, KM Anderson, CL AF Tridandapani, S Lyden, TW Smith, JL Carter, JE Coggeshall, KM Anderson, CL TI The adapter protein LAT enhances Fc gamma receptor-mediated signal transduction in myeloid cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID IMMUNOGLOBULIN-E RECEPTOR; TYROSINE PHOSPHORYLATION; HIGH-AFFINITY; PHOSPHATIDYLINOSITOL 3-KINASE; PHOSPHOINOSITIDE 3-KINASE; ANTIGEN RECEPTOR; MEMBRANE DOMAINS; CROSS-LINKING; IGE RECEPTOR; LIPID RAFTS AB Fc gamma R clustering in monocytes initiates a cascade of signaling events that culminate in biological responses such as phagocytosis, production of inflammatory cytokines, and generation of reactive oxygen species. We have identified and determined the function of the adapter protein linker of activation of T cell (LAT) in Fc gamma R-mediated signaling and function. Clustering of Fc gamma Rs on the human monocytic cell line, THP-1, induces phosphorylation of a major 36-kDa protein which immunoreacts with anti-LAT antisera, Our data indicate that although both the 36-kDa and 38-kDa isoforms of LAT are expressed in THP-1 and U937 human monocytic cells, Fc gamma R clustering induces phosphorylation of the 36-kDa isoform only. Co-immunoprecipitation experiments revealed a constitutive association of p36 LAT with both Fc gamma RI and Fc gamma RIIa immunoprecipitates, and an activation-induced association of LAT with PLC gamma 1, Grb2, and the p85 subunit of phosphatidylinositol 3-kinase. Transient transfection experiments in COS-7 cells indicated that overexpression of a wild type but not a dominant-negative LAT, that is incapable of binding to p85, enhances phagocytosis by Fc gamma RI. Furthermore, bone marrow-derived macrophages from LAT-deficient mice displayed reduced phagocytic efficiency in comparison to the macrophages from wild-type mice. Thus, we conclude that p36 LAT serves to enhance Fc gamma R-induced signal transduction in myeloid cells. C1 Ohio State Univ, Coll Med, Davis Res Ctr 2054, Dept Internal Med, Columbus, OH 43210 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20877 USA. Oklahoma Med Res Fdn, Dept Immunobiol & Canc, Oklahoma City, OK 73104 USA. RP Anderson, CL (reprint author), Ohio State Univ, Coll Med, Davis Res Ctr 2054, Dept Internal Med, 480 W 9th Ave, Columbus, OH 43210 USA. FU NCI NIH HHS [CA44983]; NICHD NIH HHS [HD35121] NR 47 TC 49 Z9 54 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20480 EP 20487 DI 10.1074/jbc.M909462199 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500037 PM 10781611 ER PT J AU Yang, Y Hwang, CK D'Souza, UM Lee, SH Junn, E Mouradian, MM AF Yang, Y Hwang, CK D'Souza, UM Lee, SH Junn, E Mouradian, MM TI Three-amino acid extension loop homeodomain proteins Meis2 and TGIF differentially regulate transcription SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DNA-BINDING SPECIFICITY; DOPAMINE-RECEPTOR GENE; HOMEOBOX GENE; RETINOIC ACID; MOLECULAR-CLONING; THYROID-HORMONE; EXPRESSION; PROMOTER; EXTRADENTICLE; REGION AB Three-amino acid extension loop (TALE) homeobox proteins are highly conserved transcription regulators. me report that two members of this family, Meis2 and TGIF, which frequently have overlapping consensus binding sites on complementary DNA strands in opposite orientations, can function competitively. For example, in the D-1A gene, which encodes the predominant dopamine receptor in the striatum, Meis2 and TGIF bind to the activator sequence ACT (-1174 to -1154) and regulate transcription differentially in a cell type-specific manner. Among the five cloned splice variants of Meis2, isoforms Meis2a-d activate the D-1A promoter in most cell types tested, whereas TGIF competes with Meis2 binding to DNA and represses Meis2-induced transcription activation, Consequently, Meis2 cannot activate the D-1A promoter in a cell that has abundant TGIF expression. The Meis2 message is highly co-localized with the D-1A message in adult striatal neurons, whereas TGIF is barely detectable in the adult brain. Our observations provide in vitro and in vivo evidence that Meis2 and TGIF differentially regulate their target genes. Thus, the delicate ratio between Meis2 and TGIF expression in a given cell type determines the cell-specific expression of the D-1A gene. We also found that splice variant Meis2e, which has a truncated homeodomain, cannot bind to the D-1A ACT sequence or activate transcription. However, Meis2e is an effective dominant negative regulator by blocking Meis2d-induced transcription activation. Thus, truncated homeoproteins with no DNA binding domains can have important regulatory functions. C1 NINDS, NIH, Expt Therapeut Branch, Genet Pharmacol Unit, Bethesda, MD 20892 USA. RP Mouradian, MM (reprint author), NINDS, NIH, Expt Therapeut Branch, Genet Pharmacol Unit, 10 Ctr Dr,MSC 1406, Bethesda, MD 20892 USA. OI Mouradian, M. Maral/0000-0002-9937-412X NR 41 TC 65 Z9 71 U1 1 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20734 EP 20741 DI 10.1074/jbc.M908382199 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500072 PM 10764806 ER PT J AU Kojima, T Morikawa, Y Copeland, NG Gilbert, DJ Jenkins, NA Senba, E Kitamura, Y AF Kojima, T Morikawa, Y Copeland, NG Gilbert, DJ Jenkins, NA Senba, E Kitamura, Y TI TROY, a newly identified member of the tumor necrosis factor receptor superfamily, exhibits a homology with edar and is expressed in embryonic skin and hair follicles SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CYTOTOXIC LIGAND TRAIL; P55 TNF RECEPTOR; NF-KAPPA-B; MOLECULAR-CLONING; SURFACE-ANTIGEN; VIRAL-PROTEINS; FAMILY; CD40; DOMAIN; ACTIVATION AB In a signal sequence trap screening of the murine brain, we identified a new member of the tumor necrosis factor receptor superfamily designated TROY. TROY is a type I membrane protein of 416 amino acids with characteristic cysteine-rich motifs in the extracellular domain and a tumor necrosis factor receptor-associated factor (TRAF) 2 binding sequence in the cytoplasmic domain of 223 amino acids. In fact, activation of nuclear factor kappa B was induced by the overexpression of TROY and inhibited by dominant negative forms of TRAF2, TRAF5, and TRAF6, indicating that TRAFs and nuclear factor kappa B are involved in the signal transduction of TROY. We also cloned a cDNA for a human counterpart, which showed a 75% homology with mouse TROY at the amino acid level. The extracellular domain of TROY exhibits an extensive homology with that of Edar, a receptor that specifies hair follicle fate. TROY mRNA is strongly expressed in brain and embryo and moderately expressed in the heart, lung, and liver but not the spleen. In the embryo, the expression level is particularly strong in the skin. Interestingly, in situ hybridization analysis of the embryo showed that TROY mRNA was exclusively expressed in the epithelium of many tissues. On the other hand, in neonatal mice, TROY is expressed in hair follicles Like Edar as well as in the cerebrum, suggesting pleiotropic functions of TROY in development as well as in the adult mice. The Troy gene is located near the waved coat (Wc) locus, a mutant related to abnormalities in skin and hair. C1 Univ Tokyo, Inst Med Sci, Dept Hematopoiet Factors, Minato Ku, Tokyo 1088639, Japan. Chugai Res Inst Mol Med Inc, Cytokine Res Program, Ibaraki, Osaka 3004101, Japan. Wakayama Med Coll, Dept Anat & Neurobiol, Wakayama 6408509, Japan. NCI, Frederick Canc Res & Dev Ctr, Mouse Canc Genet Program, Frederick, MD 21702 USA. RP Kitamura, Y (reprint author), Univ Tokyo, Inst Med Sci, Dept Hematopoiet Factors, Minato Ku, 4-6-1 Shirokanedai, Tokyo 1088639, Japan. NR 54 TC 84 Z9 90 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20742 EP 20747 DI 10.1074/jbc.M002691200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500073 PM 10764796 ER PT J AU Zhang, ZP Teng, CT AF Zhang, ZP Teng, CT TI Estrogen receptor-related receptor alpha 1 interacts with coactivator and constitutively activates the estrogen response elements of the human lactoferrin gene SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID THYROID-HORMONE RECEPTOR; ERR-1 ORPHAN RECEPTOR; NUCLEAR RECEPTORS; TRANSCRIPTIONAL ACTIVATION; CHROMOSOMAL LOCALIZATION; HISTONE DEACETYLASE; MOUSE; PROMOTER; EXPRESSION; BINDING AB The human estrogen receptor-related receptor (ERR alpha 1, NR3B1a) was shown to bind a steroidogenic factor binding element (SFRE), TCAAGGTCATC, 26 base pairs upstream from the estrogen response element (ERE) of the human lactoferrin gene promoter, A mutation made at SFRE significantly reduced estrogen-dependent transcription from the lactoferrin ERE in human endometrial cells. In this study, we demonstrated that ERR alpha 1 binds both SFRE and ERE elements and constitutively transactivates the lactoferrin gene promoter. In DNase I footprinting protection analysis, both SFRE and ERE regions were protected by glutathione S-transferase-ERR alpha 1 fusion protein. The receptor formed two protein-DNA complexes with either SFRE or ERE in electrophoresis mobility shift assay. Homodimerization of ERR alpha 1 was confirmed with the mammalian two-hybrid system. ERR alpha 1 activates reporter constructs containing various types of estrogen response elements in endometrial and non-endometrial cells in transient transfection experiments. Overexpressing the coactivator, SRC1a or GRIP1, further enhances ERR alpha 1-induced transcriptional activity. We demonstrated that the AF2 domain of ERR alpha 1 is essential for the transactivation function and that deletion or mutation at this region abrogates the activation capability. Protein-protein interaction between the SRC1a and ERR alpha 1 C terminus was confirmed with a GST glutathione S-transferase "pull-down" assay. When comparing ERR alpha 1 and the estrogen receptor alpha (ER alpha) in many of the experiments, we found that ER alpha can also bind SFRE of the lactoferrin gene and transactivate the promoter activity in a ligand-dependent manner. The present study demonstrated that ERR alpha 1 binds similar DNA elements as ER alpha and confers its transactivation function constitutively. Therefore, ERR alpha 1 may actively modulate the estrogen response of lactoferrin gene as well as other estrogen-responsive genes. C1 NIEHS, Reprod & Dev Toxicol Lab, Gene Regulat Grp, NIH, Res Triangle Pk, NC 27709 USA. RP Teng, CT (reprint author), NIEHS, Reprod & Dev Toxicol Lab, Gene Regulat Grp, NIH, POB 12233,MD E-201, Res Triangle Pk, NC 27709 USA. NR 60 TC 97 Z9 103 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20837 EP 20846 DI 10.1074/jbc.M001880200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500087 PM 10779508 ER PT J AU Fu, MF Wang, CG Reutens, AT Wang, J Angeletti, RH Siconolfi-Baez, L Ogryzko, V Avantaggiati, ML Pestell, RG AF Fu, MF Wang, CG Reutens, AT Wang, J Angeletti, RH Siconolfi-Baez, L Ogryzko, V Avantaggiati, ML Pestell, RG TI p300 and p300/cAMP-response element-binding protein-associated factor acetylate the androgen receptor at sites governing hormone-dependent transactivation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TRANSCRIPTION FACTOR GATA-1; ADENOVIRAL ONCOPROTEIN E1A; PROSTATE-SPECIFIC ANTIGEN; HISTONE ACETYLTRANSFERASES; COACTIVATORS P300; GENE ACTIVATION; DNA-DAMAGE; IN-VIVO; CBP; PCAF AB The androgen receptor (AR) is a sequence-specific DNA-binding protein that plays a key role in prostate cancer cellular proliferation by dihydrotestosterone and the induction of secondary sexual characteristics. In this study me demonstrate that the AR can be modified by acetylation in vitro and in vivo, p300 and p300/cAMP-response element-binding protein acetylated the AR at a highly conserved lysine-rich motif carboxyl-terminal to the zinc finger DNA-binding domain. [C-14]acetate-labeling experiments demonstrated that AR acetylation by p300 in cultured cells requires the same residues identified in vitro. Point mutation of the AR acetylation site (K632A/K633A) abrogated dihydrotestosterone-dependent transactivation of the AR in cultured cells. Mutation of the p300 CH3 region or the p300/cAMP-response element-binding protein histone acetylase domain reduced ligand-dependent AR function. The identification of the AR as a direct target of histone acetyltransferase co-activators has important implications for targeting inhibitors of AR function. C1 Yeshiva Univ Albert Einstein Coll Med, Albert Einstein Canc Ctr, Dept Dev & Mol Biol, Bronx, NY 10461 USA. NICHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. Roswell Pk Canc Inst, Grace Canc Drug Ctr, Dept Pharmacol & Therapeut, Buffalo, NY 14263 USA. RP Pestell, RG (reprint author), Yeshiva Univ Albert Einstein Coll Med, Albert Einstein Canc Ctr, Dept Dev & Mol Biol, 1300 Morris Pk Ave,Chanin 302, Bronx, NY 10461 USA. RI Ogryzko, Vasily/M-6665-2015 OI Ogryzko, Vasily/0000-0002-8548-1389 FU NCI NIH HHS [5-P30-CA13330-26, R01CA70897, R01CA75503] NR 49 TC 256 Z9 257 U1 2 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUL 7 PY 2000 VL 275 IS 27 BP 20853 EP 20860 DI 10.1074/jbc.M000660200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 332QG UT WOS:000088084500089 PM 10779504 ER PT J AU Gao, HQ Boyer, PL Sarafianos, SG Arnold, E Hughes, SH AF Gao, HQ Boyer, PL Sarafianos, SG Arnold, E Hughes, SH TI The role of steric hindrance in 3TC resistance of human immunodeficiency virus type-1 reverse transcriptase SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE HIV-1; reverse transcriptase; 3TC; drug resistance; RNase H ID ANTIBODY FAB FRAGMENT; DNA STRAND TRANSFER; RNASE-H; ANGSTROM RESOLUTION; HIV-1 RT; POLYMERASE; COMPLEX; PROCESSIVITY; MUTATIONS; MUTAGENESIS AB Treating HIV infections with drugs that block viral replication selects for drug-resistant strains of the virus. Particular inhibitors select characteristic resistance mutations. In the case of the nucleoside analogs 3TC and FTC, resistant viruses are selected with mutations at amino acid residue 184 of reverse transcriptase (RT). The initial change is usually to M184I; this virus is rapidly replaced by a variant carrying the mutation M184V. 3TC and ETC are taken up by cells and converted into 3TCTP and FTCTP. The triphosphate forms of these nucleoside analogs are incorporated into DNA by HIV-1 RT and act as chain terminators. Both of the mutations, M184I and M184V, provide very high levels of resistance in vivo; purified HIV-1 RT carrying M184V and M184I also shows resistance to 3TCTP and FTCTP in in vitro polymerase assays. Amino acid M184 is part of the dNTP binding site of HIV-1 RT. Structural studies suggest that the mechanism of resistance of HIV-1 RTs carrying the M184V or M184I mutation involves steric hindrance, which could either completely block the binding of 3TCTP and FTCTP or allow binding of these nucleoside triphosphate molecules but only in a configuration that would prevent incorporation. The available kinetic data are ambiguous: one group has reported that the primary effect of the mutations is at the level of 3TCTP binding; another, at the level of incorporation. We have approached this problem using assays that monitor the ability of HIV-1 RT to undergo a conformational change upon binding a dNTP. These studies show that both wild-type RT and the drug-resistant variants can bind 3TCTP at the polymerase active site; however, the binding to M184V and M184I is somewhat weaker and is sensitive to salt. We propose that the drug-resistant variants bind 3TCTP in a strained configuration that is salt-sensitive and is not catalytically competent. C1 NCI, HIV Drug Resistance Program, FCRDC, ABL Basic Res Program, Ft Detrick, 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, FCRDC, ABL Basic Res Program, POB B,Bldg 539,Room 130A, Ft Detrick, MD 21702 USA. OI Sarafianos, Stefan G/0000-0002-5840-154X FU NIAID NIH HHS [AI09578, AI27690] NR 29 TC 95 Z9 95 U1 0 U2 8 PU ACADEMIC PRESS 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 7 PY 2000 VL 300 IS 2 BP 403 EP 418 DI 10.1006/jmbi.2000.3823 PG 16 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 333HQ UT WOS:000088125200012 PM 10873473 ER PT J AU Kim, SN Kim, SG Park, JH Lee, MA Park, SD Cho-Chung, YS Hong, SH AF Kim, SN Kim, SG Park, JH Lee, MA Park, SD Cho-Chung, YS Hong, SH TI Dual anticancer activity of 8-Cl-cAMP: Inhibition of cell proliferation and induction of apoptotic cell death SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE 8-Cl-cAMP; inhibition of cell proliferation; apoptosis; anticancer activity ID DEPENDENT PROTEIN-KINASE; RII(BETA) REGULATORY SUBUNIT; HUMAN NEUROBLASTOMA-CELLS; HUMAN CANCER-CELLS; RI-ALPHA-SUBUNIT; GROWTH-INHIBITION; BCL-2 EXPRESSION; DRUG-RESISTANCE; GENE-EXPRESSION; DOWN-REGULATION AB 8-Cl-cAMP induces apoptotic cell death in human cancer cells. To look at this more closely, we examined the changes in the levels of Bcl-2 family proteins during 8-Cl-cAMP-induced apoptosis of SH-SY5Y human neuroblastoma cells. Following the treatment with 8-Cl-cAMP, Bcl-2 was transiently down-regulated and Bad was increased continuously up to day 5. In addition, overexpression of Bcl-2 efficiently blocked the 8-Cl-cAMP-induced apoptosis, suggesting Bcl-2 family proteins may be involved in the 8-Cl-cAMP-induced apoptosis. The contribution of the apoptotic cell death and the inhibition of cell proliferation in the 8-Cl-cAMP-induced growth inhibition was closely monitored in the Bcl-2-overexpressing cells. Though the apoptosis was reduced significantly, no significant difference was observed in the inhibition of cell proliferation up to day 2 of 8-Cl-cAMP treatment. These results suggest that 8-Cl-cAMP exerts anticancer activity by two distinct mechanisms, i.e., through the inhibition of cell proliferation as well as the induction of apoptosis. Supporting this notion was the observations that (1) suppression of apoptosis by zVAD did not abrogate 8-Cl-cAMP-induced inhibition of cell proliferation, and (2) 8-Cl-cAMP did not show additive inhibition of cell proliferation in RII beta-overexpressing cells. (C) 2000 Academic Press. C1 Seoul Natl Univ, Inst Mol Biol & Genet, Cell Biol Lab, Seoul 151742, South Korea. Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea. Ajou Univ, Sch Med, Brain Dis Res Ctr, Suwon 442749, South Korea. NCI, Cellular Biochem Sect, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. RP Hong, SH (reprint author), Seoul Natl Univ, Inst Mol Biol & Genet, Cell Biol Lab, Seoul 151742, South Korea. NR 34 TC 22 Z9 24 U1 0 U2 1 PU ACADEMIC PRESS INC 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 5 PY 2000 VL 273 IS 2 BP 404 EP 410 DI 10.1006/bbrc.2000.2949 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 333JZ UT WOS:000088128300003 PM 10873618 ER PT J AU Waragai, M Junn, E Kajikawa, M Takeuchi, S Kanazawa, I Shibata, M Mouradian, MM Okazawa, H AF Waragai, M Junn, E Kajikawa, M Takeuchi, S Kanazawa, I Shibata, M Mouradian, MM Okazawa, H TI PQBP-1/Npw38, a nuclear protein binding to the polyglutamine tract, interacts with US-15kD/dim1p via the carboxyl-terminal domain SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID CELL-DEATH; CYCLIN D1; TRANSCRIPTION; ACTIVATION; INDUCTION AB PQBP-1 was identified as a binding protein to the polyglutamine tract present in various transcription-related factors and causative genes for neurodegenerative disorders. This novel gene contains at least two functional domains, WW domain and carboxyl-terminal domain (CTD), strictly conserved beyond species. Although human PQBP-1 additionally contains the polar amino acid-rich domain by which it binds to the polyglutamine tract, genuine physiological function(s) have not been clarified. In this study, we showed that U5-15kD, human homologue of fission yeast dim1p, is a partner molecule of PQBP-1 binding to CTD. This finding suggests physiological functions of PQBP-1 in splicing, cell cycle, and ubiquitination, through which we can speculate the pathological roles of PQBP-1 in triplet repeat diseases. (C) 2000 Academic Press. C1 Univ Tokyo, Grad Sch Med, Dept Neurol, Bunkyo Ku, Tokyo 1138655, Japan. NINDS, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. Med & Biol Labs Co Ltd, Dept Pharmaceut Dev, Nagano 3960002, Japan. RP Okazawa, H (reprint author), Univ Tokyo, Grad Sch Med, Dept Neurol, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1138655, Japan. NR 14 TC 53 Z9 54 U1 0 U2 1 PU ACADEMIC PRESS INC 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 5 PY 2000 VL 273 IS 2 BP 592 EP 595 DI 10.1006/bbrc.2000.2992 PG 4 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 333JZ UT WOS:000088128300035 PM 10873650 ER PT J AU Chen, HD Huang, YK Ault, K Wong, GW Lin, MCM Chen, HC Kung, HF AF Chen, HD Huang, YK Ault, K Wong, GW Lin, MCM Chen, HC Kung, HF TI Molecular basis for differing antineurogenic effects of GATA-1a and GATA-1b in Xenopus SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE transcription factor; chimeras; untranslated regions; erythropoiesis; neurogenesis ID TRANSCRIPTION FACTOR GATA-1; DNA-BINDING FACTOR; DIFFERENTIAL REGULATION; EXPRESSION; GENES; NEURALIZATION AB The erythroid transcription factor GATA-1 in Xenopus has been cloned as a pair of presumably duplicated genes designated as xGATA-1a and xGATA-1b. Although both xGATA-1a and xGATA-1b are able to stimulate erythropoiesis, only xGATA-1b is capable of inhibiting neurogenesis in Xenopus embryos. Chimeras of these two genes were constructed by permuting coding and untranslated regions (UTR) on both ends of these two xGATA-1, and their neurogenesis-inhibitory effects were studied. These results reveal that (1) sequence variations between the coding regions alone do not account for the neurogenesis effect; (2) 3' UTR of xGATA-1a causes the loss of the neurogenesis inhibition of xGATA-1b; (3) 3' UTR of xGATA-1b is essential to inhibit neurogenesis. In addition, the presence of either UTR does not affect the stability of the mRNA in vitro. These observations suggest the influence of 3' UTR in xGATA-1 on the inhibition of neurogenesis. (C) 2000 Academic Press. C1 NICHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. NCI, Lab Biochem Physiol, Div Basic Sci, FCRDC,NIH, Frederick, MD 21702 USA. Univ Hong Kong, Inst Mol Biol, Hong Kong, Hong Kong, Peoples R China. RP Chen, HD (reprint author), NICHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. NR 21 TC 1 Z9 1 U1 0 U2 0 PU ACADEMIC PRESS INC 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 5 PY 2000 VL 273 IS 2 BP 614 EP 620 DI 10.1006/bbrc.2000.2988 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 333JZ UT WOS:000088128300039 PM 10873654 ER PT J AU Foncea, R Galvez, A Perez, V Morales, MP Calixto, A Melendez, J Gonzalez-Jara, F Diaz-Araya, G Sapag-Hagar, M Sugden, PH LeRoith, D Lavandero, S AF Foncea, R Galvez, A Perez, V Morales, MP Calixto, A Melendez, J Gonzalez-Jara, F Diaz-Araya, G Sapag-Hagar, M Sugden, PH LeRoith, D Lavandero, S TI Extracellular regulated kinase, but not protein kinase C, is an antiapoptotic signal of insulin-like growth factor-1 on cultured cardiac myocytes SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE insulin-like growth factor-1; protein kinases; signal transduction; apoptosis; cardiac myocytes ID RENIN-ANGIOTENSIN SYSTEM; RAT VENTRICULAR MYOCYTES; I GENE-EXPRESSION; TRANSGENIC MICE; POTENTIAL ROLE; HEART-FAILURE; IGF-I; HYPERTROPHY; APOPTOSIS; TRANSDUCTION AB This study aims to elucidate the signaling pathway for insulin-like growth factor-1 (IGF-1) in cultured neonatal rat cardiomyocytes and particularly the role of IGF-1 in cardiac apoptosis, IGF-1 stimulated polyphosphoinositide turnover, translocation of protein kinase C (PKC) isoforms (alpha, epsilon, and delta) from the soluble to the particulate fraction, activation of phospholipid-dependent and Ca2+-, phospholipid-dependent PKC, and activation of the extracellular-regulated kinase (ERK), IGF-1 attenuated sorbitol-induced cardiomyocyte viability and nuclear DNA fragmentation, These antiapoptotic effects of IGF-1 were blocked by PD098059 (an MEK inhibitor) but not by bisindolylmaleimide I (BIM, a specific PKC inhibitor), The ERK pathway may therefore be an important component in the mechanism whereby IGF-1 exerts its antiapoptotic effect on the cardiomyocyte, (C) 2000 Academic Press. C1 Univ Chile, Fac Ciencias Quim & Farmaceut, Dept Bioquim & Biol Mol, Santiago 6640750, Chile. Natl Heart & Lung Inst, Imperial Coll Sch Med, London SW3 6LY, England. NIDDK, Clin Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Lavandero, S (reprint author), Univ Chile, Fac Ciencias Quim & Farmaceut, Dept Bioquim & Biol Mol, Olivos 1007, Santiago 6640750, Chile. OI Lavandero, Sergio/0000-0003-4258-1483 NR 58 TC 29 Z9 31 U1 0 U2 2 PU ACADEMIC PRESS INC 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 5 PY 2000 VL 273 IS 2 BP 736 EP 744 DI 10.1006/bbrc.2000.3008 PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 333JZ UT WOS:000088128300058 PM 10873673 ER PT J AU Tjandra, N Tate, S Ono, A Kainosho, M Bax, A AF Tjandra, N Tate, S Ono, A Kainosho, M Bax, A TI The NMR structure of a DNA dodecamer in an aqueous dilute liquid crystalline phase SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID NUCLEAR-MAGNETIC-RESONANCE; RESTRAINED-MOLECULAR-DYNAMICS; RESIDUAL DIPOLAR COUPLINGS; EXCHANGEABLE PROTON RESONANCES; B-DNA; ORIENTED MACROMOLECULES; CONFORMATIONAL-ANALYSIS; HYDROGEN-BONDS; PSEUDOCONTACT SHIFTS; ENERGY MINIMIZATION AB The solution structure of the DNA dodecamer d(CGCGAATCCCG)(2) has been studied in an aqueous liquid crystalline medium containing 5% w/v bicelles. These phospholipid particles impose a small degree of orientation on the DNA duplex molecules with respect to the magnetic field and permit the measurement of dipolar interactions. Experiments were carried out on several samples with different isotopic labeling patterns, including two complementary samples, in which half of the nucleotides were uniformly enriched with C-13 and deuterated at the H2 " and H5' positions. From this, 198 C-13-H-1 and 10 N-15-H-1 one-bond dipolar coupling restraints were derived, in addition to 200 approximate H-1-H-1 dipolar coupling and 162 structurally meaningful NOE restraints. Although loose empirical restraints for the phosphodiester backbone torsion angles were essential for obtaining structures that satisfy all experimental data, they do not contribute to the energetic penalty function of the final minimized structures. Except for additional regular Watson-Crick hydrogen bond restraints and standard van der Waals and electrostatic terms used in the molecular dynamics-based structure calculation, the structure is determined primarily by the dipolar couplings. The final structure is highly regular, without any significant bending or kinks, and with C2'-endo/C1'-exo sugar puckers corresponding to regular a-form DNA. Most local parameters, including sugar puckers, glycosyl torsion angles, and propeller twists, are also tightly determined by the NMR data. The precision of the determined structures is limited primarily by the uncertainty in the exact magnitude and rhombicity of the alignment tensor. This causes considerable spread in parameters such as the degree of base-pair opening and the width of the minor groove, which are relatively sensitive to the alignment tensor values used. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. Japan Adv Inst Sci & Technol, Ctr New Mat, Tatsunokuchi, Ishikawa 9231292, Japan. Tokyo Metropolitan Univ, Hachioji, Tokyo 1920397, Japan. NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Tjandra, N (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 3-418, Bethesda, MD 20892 USA. NR 90 TC 172 Z9 174 U1 3 U2 20 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 5 PY 2000 VL 122 IS 26 BP 6190 EP 6200 DI 10.1021/ja000324n PG 11 WC Chemistry, Multidisciplinary SC Chemistry GA 333JF UT WOS:000088126600008 ER PT J AU Rostovtseva, TK Liu, TT Colombini, M Parsegian, VA Bezrukov, SM AF Rostovtseva, TK Liu, TT Colombini, M Parsegian, VA Bezrukov, SM TI Positive cooperativity without domains or subunits in a monomeric membrane channel SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID OUTER MITOCHONDRIAL-MEMBRANE; ION-CHANNEL; NEUROSPORA-CRASSA; VDAC; BINDING; PH; ACTIVATION; PROTEIN; NUMBER; ARRAYS AB The monomeric VDAC channel shows an accelerated pH titration of its transport properties with a Hill coefficient of about 2. This manifests itself as a sharp peak in conductance noise as well as a fast change in channel selectivity with pH. On the basis of the known structure of this channel, we propose that this cooperativity arises from a mechanically linked mobile pair of ionizable groups. Concerted movement of these groups between two states changes the distance from nearby electrostatic charge to influence the pK of the groups. This model of pH-dependent motion produces positive cooperative behavior that fits the observations without need for subunits or identifiable domains within the protein. The mathematical formalism has never required such domains, but these are generally considered an essential part of cooperative behavior in proteins. The present proposal reduces the size of a cooperative unit to a minimum, extending the limits of what is perceived to be possible. Together with large-scale conformational transitions, these subtle cooperative structural changes may allow proteins to adapt, with high sensitivity, to changes in their environment. They might also be relatively easy to engineer into a protein. C1 Univ Maryland, Dept Biol, College Pk, MD 20742 USA. NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. RP Colombini, M (reprint author), Univ Maryland, Dept Biol, College Pk, MD 20742 USA. RI Colombini, Marco/A-1540-2014 NR 26 TC 38 Z9 39 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 5 PY 2000 VL 97 IS 14 BP 7819 EP 7822 DI 10.1073/pnas.140115397 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400031 PM 10859360 ER PT J AU Wilson, SM Yip, R Swing, DA O'Sullivan, TN Zhang, Y Novak, EK Swank, RT Russell, LB Copeland, NG Jenkins, NA AF Wilson, SM Yip, R Swing, DA O'Sullivan, TN Zhang, Y Novak, EK Swank, RT Russell, LB Copeland, NG Jenkins, NA TI A mutation in Rab27a causes the vesicle transport defects observed in ashen mice SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HERMANSKY-PUDLAK-SYNDROME; COAT-COLOR PHENOTYPE; PALE EAR EP; MYOSIN-V; MELANOSOME DISTRIBUTION; PURKINJE-CELLS; GENE ENCODES; MUTANT MICE; MOUSE; PROTEIN AB The dilute (d), leaden (ln), and ashen (ash) mutations provide a unique model system for studying vesicle transport in mammals. All three mutations produce a lightened coat color because of defects in pigment granule transport. In addition, all three mutations are suppressed by the semidominant dilute-suppressor (dsu), providing genetic evidence that these mutations function in the same or overlapping transport pathways, Previous studies showed that d encodes a major vesicle transport motor, myosin-VA, which is mutated in Griscelli syndrome patients. Here, using positional cloning and bacterial artificial chromosome rescue, we show that ash encodes Rab27a, Rab GTPases represent the largest branch of the p21 Ras superfamily and are recognized as key players in vesicular transport and organelle dynamics in eukaryotic cells. We also show that ash mice have platelet defects resulting in increased bleeding times and a reduction in the number of platelet dense granules. These defects have not been reported for d and In mice. Collectively, our studies identify Rab27a as a critical gene for organelle-specific protein trafficking in melanocytes and platelets and suggest that Rab27a functions in both MyoVa dependent and independent pathways. C1 NCI, Mouse Canc Genet Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Roswell Pk Canc Inst, Dept Mol & Cell Biol, Buffalo, NY 14263 USA. Oak Ridge Natl Lab, Div Biol, Oak Ridge, TN 37831 USA. RP Jenkins, NA (reprint author), NCI, Mouse Canc Genet Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RI Yip, Richard/J-5289-2012 FU NCI NIH HHS [CA16056, P30 CA016056]; NEI NIH HHS [EY12104, R01 EY012104]; NHLBI NIH HHS [HL31698, R01 HL031698] NR 50 TC 282 Z9 287 U1 2 U2 4 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 5 PY 2000 VL 97 IS 14 BP 7933 EP 7938 DI 10.1073/pnas.140212797 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400051 PM 10859366 ER PT J AU Santamarina-Fojo, S Peterson, K Knapper, C Qiu, Y Freeman, L Cheng, JF Osorio, J Remaley, A Yang, XP Haudenschild, C Prades, C Chimini, G Blackmon, E Francois, T Duverger, N Rubin, EM Rosier, M Denefle, P Fredrickson, DS Brewer, HB AF Santamarina-Fojo, S Peterson, K Knapper, C Qiu, Y Freeman, L Cheng, JF Osorio, J Remaley, A Yang, XP Haudenschild, C Prades, C Chimini, G Blackmon, E Francois, T Duverger, N Rubin, EM Rosier, M Denefle, P Fredrickson, DS Brewer, HB TI Complete genomic sequence of the human ABCA1 gene: Analysis of the human and mouse ATP-binding cassette A promoter SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE high density lipoproteins; atherosclerosis; cholesterol efflux ID APOLIPOPROTEIN-A-I; DENSITY-LIPOPROTEIN DEFICIENCY; REVERSE CHOLESTEROL TRANSPORT; TRANSFER PROTEIN GENE; TANGIER-DISEASE; TRANSCRIPTION FACTORS; MOLECULAR MECHANISMS; CELLULAR CHOLESTEROL; NUCLEAR FACTOR-3; RESPONSE ELEMENT AB The ABCA1 gene, a member of the ATP-binding cassette A (ABCA1) transporter superfamily, encodes a membrane protein that facilitates the cellular efflux of cholesterol and phospholipids. Mutations in ABCA1 lead to familiar high density lipoprotein deficiency and Tangier disease. We report the complete human ABCA1 gene sequence, including 1,453 bp of the promoter, 146,581 bp of introns and exons, and 1 kb of the 3' flanking region. The ABCA1 gene spans 149 kb and comprises 50 exons. Sixty-two repetitive Alu sequences were identified in introns 1-49. The transcription start site is 315 bp upstream of a newly identified initiation methionine codon and encodes an ORF of 6,783 bp. Thus, the ABCA1 protein is comprised of 2,261 aa. Analysis of the 1,453 bp 5' upstream of the transcriptional start site reveals multiple binding sites for transcription factors with roles in lipid metabolism. Comparative analysis of the mouse and human ABCA1 promoter sequences identified specific regulatory elements, which are evolutionarily conserved. The human ABCA1 promoter fragment -200 to -80 bp that contains binding motifs for SP1, SP3, E-box, and AP1 modulates cellular cholesterol and cAMP regulation of ABCA1 gene expression. These combined findings provide insights into ABCA1-mediated regulation of cellular cholesterol metabolism and will facilitate the identification of new pharmacologic agents for the treatment of atherosclerosis in humans. C1 NHLBI, NIH, Mol Dis Branch, Bethesda, MD 20892 USA. NIH, Dept Clin Pathol, Bethesda, MD 20892 USA. Ctr Immunol Marseille Luminy, F-13288 Marseille, France. Aventis Pharma, Evry Genom Ctr, F-91047 Evry, France. Univ Calif Berkeley, Lawrence Berkeley Lab, Berkeley, CA 94720 USA. RP Santamarina-Fojo, S (reprint author), NHLBI, NIH, Mol Dis Branch, 10-7N115,10 Ctr Dr, Bethesda, MD 20892 USA. NR 54 TC 147 Z9 161 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 5 PY 2000 VL 97 IS 14 BP 7987 EP 7992 DI 10.1073/pnas.97.14.7987 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400060 PM 10884428 ER PT J AU Geiszt, M Kopp, JB Varnai, P Leto, TL AF Geiszt, M Kopp, JB Varnai, P Leto, TL TI Identification of Renox, an NAD(P)H oxidase in kidney SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NADPH OXIDASE; ERYTHROPOIETIN GENE; TRANSGENIC MICE; TUBULAR CELLS; CYTOCHROME-B558; COMPONENTS; EXPRESSION; SENESCENCE; PROTEINS; BINDING AB Oxygen sensing is essential for homeostasis in all aerobic organisms, but its mechanism is poorly understood. Data suggest that a phagocytic-like NAD(P)H oxidase producing reactive oxygen species serves as a primary sensor for oxygen. We have characterized a source of superoxide anions in the kidney that we refer to as a renal NAD(P)H oxidase or Renox. Renox is homologous to gp91(phox) (91-kDa subunit of the phagocyte oxidase), the electron-transporting subunit of phagocytic NADPH oxidase. and contains all of the structural motifs considered essential for binding of heme, flavin, and nucleotide, In situ RNA hybridization revealed that renox is highly expressed at the site of erythropoietin production in the renal cortex, showing the greatest accumulation of renox mRNA in proximal convoluted tubule epithelial cells. NIH 3T3 fibroblasts overexpressing transfected Renox show increased production of superoxide and develop signs of cellular senescence. Our data suggest that Renox, as a renal source of reactive oxygen species, is a likely candidate for the oxygen sensor function regulating oxygen-dependent gene expression and may also have a role in the development of inflammatory processes in the kidney. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. NICHHD, Endocrinol & Reprod Branch, NIH, Bethesda, MD 20892 USA. RP Leto, TL (reprint author), NIAID, Host Def Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 27 TC 542 Z9 551 U1 0 U2 11 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 5 PY 2000 VL 97 IS 14 BP 8010 EP 8014 DI 10.1073/pnas.130135897 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400064 PM 10869423 ER PT J AU Zhao, WQ Cavallaro, S Gusev, P Alkon, DL AF Zhao, WQ Cavallaro, S Gusev, P Alkon, DL TI Nonreceptor tyrosine protein kinase pp60c-src in spatial learning: Synapse-specific changes in its gene expression, tyrosine phosphorylation, and protein-protein interactions SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID LONG-TERM POTENTIATION; GROWTH CONE MEMBRANES; DEVELOPING RAT-BRAIN; C-SRC; NEUROTRANSMITTER RELEASE; NMDA RECEPTORS; PRODUCT; NEURONS; BINDING; VESICLE AB c-src is a nonreceptor tyrosine protein kinase that is highly concentrated in synaptic regions, including synaptic vesicles and growth cones. Here, we report that the mRNA signal of pp60c-src is widely distributed in the rat brain with particularly high concentrations in the hippocampus. After spatial maze learning, up-regulation of c-src mRNA was observed in the CA3 region of the hippocampus, which was accompanied by increases in pp60c-src protein in hippocampal synaptosomal preparations. Training also triggered an increase in c-src protein tyrosine kinase activity that was correlated with its tyrosine dephosphorylation in the synaptic membrane fraction, After training, pp60c-src from hippocampus showed enhanced interactions with synaptic proteins such as synapsin I, synaptophysin, and the type 2 N-methyl-D-aspartate receptor, as well as the cytoskeletal protein actin, The association of pp60c-src with insulin receptor in the synaptic membrane fraction, however, was temporally decreased after training. Furthermore, in vitro results showed that Ca2+ and protein kinase C might be involved in the regulation of protein-protein interactions of pp60c-src. These results suggest, therefore, that pp60c-src participates in the regulation of hippocampal synaptic activity during learning and memory. C1 NINDS, Lab Adapt Syst, NIH, Bethesda, MD 20892 USA. CNR, Inst Bioimaging & Pathophysiol Cent Nervous Syst, I-95123 Catania, Italy. RP Zhao, WQ (reprint author), NINDS, Lab Adapt Syst, NIH, Bethesda, MD 20892 USA. RI Cavallaro, Sebastiano/F-3104-2010 OI Cavallaro, Sebastiano/0000-0001-7590-1792 NR 42 TC 49 Z9 52 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 5 PY 2000 VL 97 IS 14 BP 8098 EP 8103 DI 10.1073/pnas.97.14.8098 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400079 PM 10884433 ER PT J AU Guan, ZH Prado, A Melzig, J Heisenberg, M Nash, HA Raabe, T AF Guan, ZH Prado, A Melzig, J Heisenberg, M Nash, HA Raabe, T TI Mushroom body defect, a gene involved in the control of neuroblast proliferation in Drosophila, encodes a coiled-coil protein SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ENDOPLASMIC-RETICULUM; BRAIN MUTANTS; MELANOGASTER; KINASE; NEUROGENESIS; GENERATION; MEMBRANE; REGIONS; NEURONS; DOMAIN AB Neurogenesis relies on the establishment of the proper number and precisely controlled proliferation of neuroblasts, the neuronal precursor cells. A role for the mushroom body defect (mud) gene in both of these aspects of neuroblast behavior, as well as possible roles in other aspects of fruit fly biology, is implied by phenotypes associated with mud mutations. We have localized mud by determining the sequence change in one point mutant, identifying a predicted ORF affected by the mutation, and showing that an appropriate segment of the genome rescues mud mutant phenotypes. An analysis of mud cDNAs and a survey of mud transcripts by Northern blotting indicate that the gene is subject to differential splicing and is expressed primarily during embryogenesis but also, at lower levels, during subsequent developmental stages in a sexually dimorphic manner. The gene is predicted to encode a polypeptide without obvious homologs but with two prominent structural features, a long coiled coil that constitutes the central core of the protein and a carboxyl-terminal transmembrane domain. C1 Univ Wurzburg, Lehrstuhl Genet, Theodor Boveri Inst Biowissensch, D-97074 Wurzburg, Germany. NIMH, Mol Biol Lab, Mol Genet Sect, Bethesda, MD 20892 USA. RP Raabe, T (reprint author), Univ Wurzburg, Lehrstuhl Genet, Theodor Boveri Inst Biowissensch, D-97074 Wurzburg, Germany. RI Prado, Antonio/N-3401-2014 OI Prado, Antonio/0000-0001-9813-8566 NR 43 TC 29 Z9 30 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 5 PY 2000 VL 97 IS 14 BP 8122 EP 8127 DI 10.1073/pnas.97.14.8122 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 331ZK UT WOS:000088048400083 PM 10884435 ER PT J AU Liu, J Liu, YP Habeebu, SM Waalkes, MP Klaassen, CD AF Liu, J Liu, YP Habeebu, SM Waalkes, MP Klaassen, CD TI Chronic combined exposure to cadmium and arsenic exacerbates nephrotoxicity, particularly in metallothionein-I/II null mice SO TOXICOLOGY LA English DT Article DE cadmium; arsenite; chronic oral exposure; metallothionein; nephrotoxicity; metallothionein-I/II null mice ID INDUCED HEPATOTOXICITY; OXIDATIVE STRESS; II GENES; CARCINOGENESIS; TOXICITY; CANCER; ZINC; RATS; SENSITIVITY; METABOLISM AB Cadmium (Cd) and arsenic (As) are important inorganic toxicants in the environment. Humans certainly have the potential to be exposed to the mixtures of Cd and As, but the toxicological interactions of these inorganic mixtures are poorly defined. Metallothionein (MT) is a cysteine-rich, metal-binding protein that plays an important role in Cd detoxication, but its role in As toxicity is less certain. To examine the role of MT in Cd- and/or As-induced nephrotoxicity, MT-I/II-knockout (MT-null) mice and background-matched wild-type (WT) mice were fed CdCl2 (100 ppm Cd) in the diet, NaAsO2 (22.5 ppm As) in the drinking water, or Cd plus As for 4 months. Subsequently, nephrotoxicity was examined by morphological and biochemical techniques. Chronic exposure to Cd produced more renal toxicity than As, and the combination of Cd and As produced even more renal injury than caused by either of the chemicals given alone. In mice receiving Cd plus As, proximal tubule degeneration and atrophy, glomerular swelling and interstitial fibrosis were more severe than those produced by either inorganic. Furthermore, lack of MT rendered MT-null mice more sensitive than WT mice to the nephrotoxicity produced by chronic Cd- and/or As-exposure. MT-null mice were especially susceptible to the toxicity produced by the combination of Cd and As, as evidenced by decreased body weight, enzymuria, glucosuria, proteinuria and nephropathy. In conclusion, this study indicates that As may potentiate Cd nephrotoxicity during the long-term, combined exposure, and that intracellular MT plays a role in decreasing the nephropathy of combined exposure to Cd and As. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Comparat Carcinogenesis Lab, NCI, Res Triangle Pk, NC 27709 USA. Univ Kansas, Med Ctr, Kansas City, KS 66103 USA. RP Liu, J (reprint author), NIEHS, Comparat Carcinogenesis Lab, NCI, Mail Drop F0-09, Res Triangle Pk, NC 27709 USA. NR 46 TC 60 Z9 61 U1 2 U2 10 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 JUL 5 PY 2000 VL 147 IS 3 BP 157 EP 166 DI 10.1016/S0300-483X(00)00194-3 PG 10 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 337UR UT WOS:000088380500002 PM 10924798 ER PT J AU Hasenkrug, KJ Dittmer, U AF Hasenkrug, KJ Dittmer, U TI The role of CD4 and CD8 T cells in recovery and protection from retroviral infection: Lessons from the friend virus model SO VIROLOGY LA English DT Review ID NEUTRALIZING ANTIBODY-RESPONSE; HOST GENETIC-CONTROL; INDUCED LEUKEMIA; TYPE-1 INFECTION; MICE; LYMPHOCYTE; IMMUNITY; COMPLEX; VIREMIA; DISEASE C1 NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. Univ Wurzburg, Inst Virol, Wurzburg, Germany. RP Hasenkrug, KJ (reprint author), NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, 903 S 4th St, Hamilton, MT 59840 USA. NR 38 TC 45 Z9 45 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD JUL 5 PY 2000 VL 272 IS 2 BP 244 EP 249 DI 10.1006/viro.2000.0387 PG 6 WC Virology SC Virology GA 334AL UT WOS:000088162900002 PM 10873767 ER PT J AU Ravn, P Wasnich, RD Cizza, G AF Ravn, P Wasnich, RD Cizza, G TI Long-term prevention of bone loss - In response SO ANNALS OF INTERNAL MEDICINE LA English DT Letter ID POSTMENOPAUSAL C1 Ctr Clin & Basic Res, Ballerup, Denmark. Hawaii Osteoporosis Ctr, Honolulu, HI 96814 USA. NIMH, Bethesda, MD 20892 USA. RP Ravn, P (reprint author), Ctr Clin & Basic Res, Ballerup, Denmark. NR 5 TC 0 Z9 0 U1 0 U2 0 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 4 PY 2000 VL 133 IS 1 BP 72 EP 73 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 329TB UT WOS:000087923100012 ER PT J AU Prasad, A Narayanan, S Husain, S Padder, F Waclawiw, M Epstein, N Quyyumi, AA AF Prasad, A Narayanan, S Husain, S Padder, F Waclawiw, M Epstein, N Quyyumi, AA TI Insertion-deletion polymorphism of the ACE gene modulates reversibility of endothelial dysfunction with ACE inhibition SO CIRCULATION LA English DT Article DE atherosclerosis; genes; angiotensin; acetylcholine; coronary disease ID ANGIOTENSIN-CONVERTING-ENZYME; CORONARY-ARTERY DISEASE; ISCHEMIC-HEART-DISEASE; NITRIC-OXIDE ACTIVITY; MYOCARDIAL-INFARCTION; LINKAGE ANALYSIS; RISK-FACTORS; BRADYKININ; VASODILATION; SEGREGATION AB Background-The aim of this study was to examine whether angiotensin-converting enzyme (ACE) inhibition improves coronary endothelial dysfunction in patients with atherosclerosis and its risk factors and whether this was related to the ACE insertion-deletion (I/D) polymorphism. Methods and Results-In 56 patients with atherosclerosis or its risk factors, we studied endothelium-dependent responses with acetylcholine and endothelium-independent function with sodium nitroprusside, before and after ACE inhibition with enalaprilat. Enalaprilat did not alter either resting coronary tone or vasodilation with sodium nitroprusside. However, it potentiated the coronary microvascular and epicardial responses with acetylcholine; coronary blood flow increased from 82 +/- 7 to 90 +/- 8 mL/min (P=0.05) after enalaprilat. Patients with depressed endothelial function (P<0.001) and those with ACE DD or ID genotypes (P=0.002) but not those homozygous for the I allele had the greatest improvement by multivariate analysis. Similarly, acetylcholine-mediated epicardial vasomotion improved in segments that initially constricted (endothelial dysfunction): from - 10.1 +/- 1% to -1.4 +/- 2% (P<0.001) after enalaprilat. No augmentation was observed in segments that dilated (normal endothelial dysfunction) with acetylcholine. Patients with the D allele, hypercholesterolemia, and smokers (all P<0.05) had greater improvement. Conclusions-Acute ACE inhibition improves coronary epicardial and microvascular endothelium-dependent vasomotion in patients with atherosclerosis or its risk factors who have endothelial dysfunction and presence of the D allele. C1 NHLBI, NIH, Cardiol Branch, Bethesda, MD 20892 USA. RP Quyyumi, AA (reprint author), NHLBI, NIH, Cardiol Branch, Bldg 10,Room 7B15,10 Ctr Dr,MSC 1650, Bethesda, MD 20892 USA. NR 32 TC 44 Z9 48 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 4 PY 2000 VL 102 IS 1 BP 35 EP 41 PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 329VG UT WOS:000087928700009 PM 10880412 ER PT J AU Kitani, A Fuss, IJ Nakamura, K Schwartz, OM Usui, T Strober, W AF Kitani, A Fuss, IJ Nakamura, K Schwartz, OM Usui, T Strober, W TI Treatment of experimental (Trinitrobenzene sulfonic acid) colitis by intranasal administration of transforming growth factor (TGF)-beta 1 plasmid: TGF-beta 1-mediated suppression of T helper cell type 1 response occurs by interleukin (IL)-10 induction and IL-12 receptor beta 2 chain downregulation SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE delivery; hapten; interferon gamma; tumor necrosis factor alpha; beta-galactosidase ID INFLAMMATORY-BOWEL-DISEASE; TUMOR-NECROSIS-FACTOR; GENE-TRANSFER; INTESTINAL INFLAMMATION; MUCOSAL INFLAMMATION; SIGNAL TRANSDUCER; TRANSGENIC MICE; ORAL TOLERANCE; SCID MICE; EXPRESSION AB In this study, we show that a single intranasal dose of a plasmid encoding active transforming growth factor beta 1 (pCMV-TGF-beta 1) prevents the development of T helper cell type 1 (Th1)-mediated experimental colitis induced by the haptenating reagent, 2,4,6-trinitrobenzene sulfonic acid (TNBS). In addition, such plasmid administration abrogates TNBS colitis after it has been established, whereas, in contrast, intraperitoneal administration of rTGF-beta 1 protein does not have this effect. Intranasal pCMV-TGF-beta 1 administration leads to the expression of TGF-beta 1 mRNA ill the intestinal lamina propria and spleen for 2 wk, as well as the appearance of TGF-beta 1-producing T cells and macrophages in these tissues, and is not associated with the appearances of fibrosis. These cells cause marked suppression of interleukin (IL)-12 and interferon (IFN)-gamma production and enhancement of IL-10 production; in addition, they inhibit IL-12 receptor beta 2 (IL-12R beta 2) chain expression. Coadministration of anti-IL-10 at the time of pCMV-TCF-beta 1 administration prevents the enhancement of IL-10 production and reverses the supression of IL-12 but not IFN-gamma secretion. However, anti-IL-10 leads to increased tumor necrosis factor or production, especially in established colitis. Taken together, these studies show that TGF-P 1 inhibition of a Th1-mediated colitis is due to: (a) suppression of IL-12 secretion by IL-10 induction and (b) inhibition of IL-12 signaling via downregulation of IL-12R beta 2 chain expression. In addition, TGF-beta 1 may also have an inhibitory effect on IFN-gamma transcription. C1 NIAID, Mucosal Immun Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. NIAID, Biol Imaging Facil, NIH, Bethesda, MD 20892 USA. RP Strober, W (reprint author), Bldg 10,Rm 11N238,10 Ctr Dr, Bethesda, MD 20892 USA. NR 52 TC 131 Z9 140 U1 0 U2 1 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 3 PY 2000 VL 192 IS 1 BP 41 EP 52 DI 10.1084/jem.192.1.41 PG 12 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 334NB UT WOS:000088191600005 PM 10880525 ER PT J AU Daly, JW AF Daly, JW TI Alkylxanthines as research tools SO JOURNAL OF THE AUTONOMIC NERVOUS SYSTEM LA English DT Review DE caffeine; methylxanthines; pharmacological effects ID CYCLIC-NUCLEOTIDE PHOSPHODIESTERASE; ADENOSINE-RECEPTOR ANTAGONISTS; SMOOTH-MUSCLE CELLS; RAT SENSORY NEURONS; XANTHINE DERIVATIVES; CAFFEINE ANALOGS; CHLORIDE EFFLUX; RELEASE CHANNEL; CALCIUM-RELEASE; CA2+ RELEASE AB (1) The methylxanthine caffeine has many pharmacological effects, most of which can be Linked to blockade of adenosine receptors, inhibition of phosphodiesterases, and augmentation of calcium-dependent release of calcium from intracellular stores. (2) A variety of xanthines have been developed as potent and/or selective antagonists for adenosine receptors. (3) Several xanthines have been developed that are more potent and more selective inhibitors of cyclic nucleotide phosphodiesterase than caffeine or theophylline. (4) Caffeine remains the xanthine of choice for activation of intracellular calcium-sensitive calcium release channels although millimolar concentrations are required, which can have effects on other aspects of calcium regulation. (C) 2000 Elsevier Science BN. All rights reserved. C1 NIDDKD, Sect Pharmacodynam, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Daly, JW (reprint author), NIDDKD, Sect Pharmacodynam, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NR 103 TC 40 Z9 41 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-1838 J9 J AUTONOM NERV SYST JI J. Auton. Nerv. Syst. PD JUL 3 PY 2000 VL 81 IS 1-3 BP 44 EP 52 DI 10.1016/S0165-1838(00)00110-7 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 335FA UT WOS:000088232000008 PM 10869699 ER PT J AU Jacobson, KA Kim, YC King, BF AF Jacobson, KA Kim, YC King, BF TI In search of selective P2 receptor ligands: interaction of dihydropyridine derivatives at recombinant rat P2X(2) receptors SO JOURNAL OF THE AUTONOMIC NERVOUS SYSTEM LA English DT Article DE ion channels; oocytes; purines; dihydropyridine derivatives; potentiator ID P2Y(1) RECEPTORS; ION CHANNELS; ANTAGONISTS; ATP; POTENT; ACTIVATION; RESPONSES; NEURONS; CELLS AB 1,4-Dihydropyridines are regarded as privileged structures for drug design, i.e. they tend to bind to a wide variety of receptor sites. We have shown that upon appropriate manipulation of the substituent groups on a 1.4-dihydropyridine template, high affinity and selectivity for the A, subtype of adenosine receptors ('P1 receptors') may be attained. In the present study we have begun to extend this approach to P2 receptors which are activated by ATP and other nucleotides. Nicardipine, a representative dihydropyridine, used otherwise as an L-type calcium channel blocker, was shown to be an antagonist at recombinant rat P2X(2) (IC(50) = 25 mu M) and P2X(4) (IC(50) similar to 220 mu M) receptors expressed in Xenopus oocytes. Thus, this class of compounds represents a suitable lead for enhancement of affinity through chemical synthesis. In an attempt to modify the 1,4-dihydropyridine structure with a predicted P2 receptor recognition moiety, we have replaced one of the ester groups with a negatively charged phosphonate group. Several 4-phenyl-5-phosphonato-1,4-dihydropyridine derivatives, MRS 2154 (2,6-dimethyl), MRS 2155 (6-methyl-2-phenyl), and MRS 2156 (2-methyl-6-phenyl), were synthesized through three component condensation reactions. These derivatives were not pure antagonists of the effects of ATP at P2X(2) receptors, rather were either inactive (MRS 2156) or potentiated the effects of ATP in a concentration-dependent manner (MRS 2154 in the 0.3-10 mu M range and MRS 2155 at >1 mu M). Antagonism of the effects of ATP at P2X(2) receptor superimposed on the potentiation was also observed at >10 mu M (MRS 2154) or 0.3-1 mu M (MRS 2155). Thus, while a conventional dihydropyridine, nicardipine, was found to antagonize rat P2X(2) receptors ninefold more potently than P2X(4) receptors, the effects of novel, anionic 5-phosphonate analogues at the receptor were more complex. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. UCL, Sch Med, Autonom Neurosci Inst, London NW3 2PF, England. RP Jacobson, KA (reprint author), NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bldg 8A Rm B1A-19, Bethesda, MD 20892 USA. EM kajacobs@helix.nih.gov RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 FU Intramural NIH HHS [Z01 DK031127-01, Z99 DK999999] NR 31 TC 16 Z9 16 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-1838 J9 J AUTONOM NERV SYST JI J. Auton. Nerv. Syst. PD JUL 3 PY 2000 VL 81 IS 1-3 BP 152 EP 157 DI 10.1016/S0165-1838(00)00128-4 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 335FA UT WOS:000088232000023 PM 10869714 ER PT J AU Bezrukov, SM Winterhalter, M AF Bezrukov, SM Winterhalter, M TI Examining noise sources at the single-molecule level: 1/f noise of an open maltoporin channel SO PHYSICAL REVIEW LETTERS LA English DT Article ID SELF-ORGANIZED CRITICALITY; CURRENT FLUCTUATIONS; ESCHERICHIA-COLI; ION-CHANNEL; SALMONELLA-TYPHIMURIUM; SUGAR-TRANSPORT; RATE CONSTANTS; KINETICS; BINDING; PROTEIN AB We have studied the phenomenological origin of 1/f noise in a solute-specific bacterial inn channel, maltoporin. We show that after excision of small, but resolvable stepwise changes in the recordings of the current through a single open channel, the 1/f noise component disappears and the channel exhibits noise that is "white" below 100 Hz. Combined with results of a recent noise study of several bacterial porins, our observations suggest that 1/f noise is caused by the equilibrium conductance fluctuations related to the conformational flexibility of the channel pore structural constituents. C1 NICHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. Biozentrum, Dept Biophys Chem, Basel, Switzerland. IPBS, CNRS, Toulouse, France. RP Bezrukov, SM (reprint author), NICHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. NR 29 TC 106 Z9 106 U1 0 U2 15 PU AMERICAN PHYSICAL SOC PI COLLEGE PK PA ONE PHYSICS ELLIPSE, COLLEGE PK, MD 20740-3844 USA SN 0031-9007 J9 PHYS REV LETT JI Phys. Rev. Lett. PD JUL 3 PY 2000 VL 85 IS 1 BP 202 EP 205 DI 10.1103/PhysRevLett.85.202 PG 4 WC Physics, Multidisciplinary SC Physics GA 330JR UT WOS:000087959400051 PM 10991194 ER PT J AU Vaitukaitis, JL AF Vaitukaitis, JL TI Reviving patient-oriented research SO ACADEMIC MEDICINE LA English DT Editorial Material C1 NIH, Natl Ctr Res Resources, Bethesda, MD 20892 USA. RP Vaitukaitis, JL (reprint author), NIH, Natl Ctr Res Resources, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU HANLEY & BELFUS INC PI PHILADELPHIA PA 210 S 13TH ST, PHILADELPHIA, PA 19107 USA SN 1040-2446 J9 ACAD MED JI Acad. Med. PD JUL PY 2000 VL 75 IS 7 BP 683 EP 685 DI 10.1097/00001888-200007000-00008 PG 3 WC Education, Scientific Disciplines; Health Care Sciences & Services SC Education & Educational Research; Health Care Sciences & Services GA 337QD UT WOS:000088372400008 PM 10926017 ER PT J AU Lands, WEM AF Lands, WEM TI Peptide signaling paths related to intoxication, memory and addiction SO ADDICTION BIOLOGY LA English DT Review ID METHYL-D-ASPARTATE; PROTEIN-KINASE-C; LONG-TERM POTENTIATION; ALCOHOL-DEPENDENT PATIENTS; NMDA-RECEPTOR ANTAGONISTS; FYN TYROSINE KINASE; I ADENYLYL-CYCLASE; BEHAVIORAL SENSITIZATION; ETHANOL SENSITIVITY; TRANSIENT CALCIUM AB Many peptides bind to G protein-coupled receptors and activate intracellular signaling paths for adaptive cellular responses. The components of these paths can be affected by signals from other neurotransmitters to produce overall integrated results not easily predicted from customary a priori considerations. This intracellular cross-talk among signaling paths provides a "filter" through which long-term tonic signals affect short-term phasic signals as they progress toward the nucleus and induce long-term adaptation of gene expression which provide enduring attributes of acquired memories and addictions. Peptides of the PACAP family provide intracellular signaling that involves kinases, scaffolding interactions, Ca2+ mobilization, and gene expression to facilitate development of tolerance to alcohol and development of associative memories. The peptide-induced enhancement of NMDA receptor responses to extracellular glutamate also may increase behavioral sensitization to the low doses of alcohol that occur at the onset of each bout of drinking. Because many gene products participate in each signaling path, each behavioral response to alcohol is a polygenic process of many steps with no single gene product sufficient to interpret fully the adaptive response to alcohol. Different susceptibility of individuals to alcohol addiction may be a cumulative result of small differences among the many signaling components. Understanding this network of signals may help interpret future "magic bullets" proposed to treat addiction. C1 NIAAA, NIH, Bethesda, MD 20892 USA. RP Lands, WEM (reprint author), NIAAA, NIH, 6000 Execut Blvd,MC 7003, Bethesda, MD 20892 USA. NR 98 TC 0 Z9 0 U1 0 U2 0 PU CARFAX PUBLISHING PI BASINGSTOKE PA RANKINE RD, BASINGSTOKE RG24 8PR, HANTS, ENGLAND SN 1355-6215 J9 ADDICT BIOL JI Addict. Biol. PD JUL PY 2000 VL 5 IS 3 BP 245 EP 260 DI 10.1111/j.1369-1600.2000.tb00189.x PG 16 WC Biochemistry & Molecular Biology; Substance Abuse SC Biochemistry & Molecular Biology; Substance Abuse GA 339TB UT WOS:000088493000001 PM 20575839 ER PT J AU Ratnasinghe, D Forman, MR Tangrea, JA Qiao, YL Yao, SX Gunter, EW Barrett, MJ Giffen, CA Erozan, Y Tockman, MS Taylor, PR AF Ratnasinghe, D Forman, MR Tangrea, JA Qiao, YL Yao, SX Gunter, EW Barrett, MJ Giffen, CA Erozan, Y Tockman, MS Taylor, PR TI Serum carotenoids are associated with increased lung cancer risk among alcohol drinkers, but not among non-drinkers in a cohort of tin miners SO ALCOHOL AND ALCOHOLISM LA English DT Article ID HEALTH INTERVIEW SURVEY; BETA-CAROTENE; ALPHA-TOCOPHEROL; DIETARY-INTAKE; UNITED-STATES; TOBACCO USE; VEGETABLES; SMOKING; YUNNAN; CHINA AB To examine the association between pre-diagnostic serum carotenoid levels and lung cancer risk and the effects of alcohol intake on the carotenoid-lung cancer relationship, we conducted a case-control study in an occupational cohort from the Yunnan Tin Corporation in China. During 6 years of follow-up, 339 cases of confirmed lung cancer were diagnosed. Among these cases, those who donated pre-diagnostic blood (n = 108) were eligible for this study. For each case, two individuals alive and free of cancer at the time of case diagnosis, matched on age, sex, and dare of blood collection, were selected as controls. Serum beta-carotene (odds ratios (ORs) for tertiles: 1, 1.3, 2.0) and beta-cryptoxanthin (ORs for tertiles: 1, 1.8, 2.9) levels were positively associated with lung cancer risk after adjustment for tobacco use and radon exposure. Among alcohol drinkers, higher serum carotenoid levels were significantly associated with increased lung cancer risk (alpha-carotene OR 2.2, 95% confidence interval (CI) 1.1-4.4, beta-carotene OR 7.6, 95% CI 3.1-18.6, lutcin/zeaxanthin OR 2.3, 95% CI 1.2-6.6 and beta-cryptoxanthin OR 7.6, 95% CI 2.7-21.5). Conversely, risk estimates among non-drinkers suggest a possible protective association for higher carotenoid levels. C1 NCI, Canc Prevent Studies Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. CICAMS, Beijing, Peoples R China. Yunnan Tin Corp, Gejiu, Yunnan Province, Peoples R China. Ctr Dis Control & Prevent, NHANES Lab Biochem Anal, Atlanta, GA USA. Informat Management Serv Inc, Silver Spring, MD USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. H Lee Moffit Canc Ctr & Res Inst, Tampa, FL USA. RP Ratnasinghe, D (reprint author), NCI, Canc Prevent Studies Branch, Div Clin Sci, NIH, 6006 Execut Blvd,Suite 321, Bethesda, MD 20892 USA. RI Qiao, You-Lin/B-4139-2012 OI Qiao, You-Lin/0000-0001-6380-0871 NR 32 TC 22 Z9 22 U1 1 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0735-0414 J9 ALCOHOL ALCOHOLISM JI Alcohol Alcohol. PD JUL-AUG PY 2000 VL 35 IS 4 BP 355 EP 360 DI 10.1093/alcalc/35.4.355 PG 6 WC Substance Abuse SC Substance Abuse GA 347JT UT WOS:000088926400007 PM 10906000 ER EF