FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Domanski, MJ Epstein, A Hallstrom, A Saksena, S Zipes, DP AF Domanski, MJ Epstein, A Hallstrom, A Saksena, S Zipes, DP TI Survival of antiarrhythmic or implantable cardioverter defibrillator treated patients with varying degrees of left ventricular dysfunction who survived malignant ventricular arrhythmias SO JOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY LA English DT Article DE sudden cardiac death; implantable cardioverter defibrillator; ventricular arrhythmia ID MYOCARDIAL-INFARCTION; RANDOMIZED TRIAL; CARDIAC-ARREST; TACHYARRHYTHMIAS; THERAPY AB Introduction: The relative effectiveness of the implantable cardioverter defibrillator (ICD) and antiarrhythmic drugs (AADs) varies with left ventricular ejection fraction (LVEF). However, once an ICD or AAD treatment strategy is chosen, the degree to which the LVEF influences survival is unknown. This article addresses that question, Methods and Results: Using patient data from the Antiarrhythmics Versus Implantable Defibrillators (AVID) trial, the impact of LVEF on prognosis of patients who were treated with either an ICD or AAD was assessed. Survival within each quintile of LVEF was estimated by the method of Kaplan-Meier for patients treated with either the ICD or AADs. The Cox proportional hazards model was used to investigate the prognostic value of LVEF for estimating survival after adjustment for other baseline covariates among all patients in the subgroups treated by ICD or AAD. In the highest two quintiles of LVEF, survival was comparable in AAD-treated and ICD-treated patients. In the AAD-treated patients, higher LVEF was significantly and independently associated with survival free of all-cause mortality and arrhythmic death. In the ICD-treated patients, however, the statistical significance of the association was lost and only a trend toward greater survival was present. Death due to congestive heart failure remained independently and significantly associated with survival in both AAD-treated and ICD-treated patients. Conclusion: In patients treated with AADs but not patients treated with ICDs, survival is strongly associated with LVEF. The absence of a statistically significant association in the ICD patients is likely related to the effectiveness of the ICD in treating malignant ventricular arrhythmias, but a chance lack of association cannot be excluded. C1 NHLBI, Clin Trials Grp, Rockledge Ctr 2, Bethesda, MD 20892 USA. Univ Alabama, Sch Med, Div Cardiol, Birmingham, AL USA. Univ Washington, Div Biostat, Seattle, WA 98195 USA. Cardiovasc Inst Atlantic Hlth Syst, Passaic, NJ USA. Indiana Univ, Sch Med, Div Cardiol, Indianapolis, IN USA. RP Domanski, MJ (reprint author), NHLBI, Clin Trials Grp, Rockledge Ctr 2, 6701 Rockledge Dr,MSC 7936, Bethesda, MD 20892 USA. OI Zipes, Douglas/0000-0001-7141-6829 NR 11 TC 11 Z9 11 U1 0 U2 1 PU FUTURA PUBL CO PI ARMONK PA 135 BEDFORD RD, PO BOX 418, ARMONK, NY 10504-0418 USA SN 1045-3873 J9 J CARDIOVASC ELECTR JI J. Cardiovasc. Electrophysiol. PD JUN PY 2002 VL 13 IS 6 BP 580 EP 583 DI 10.1046/j.1540-8167.2002.00580.x PG 4 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 562NT UT WOS:000176205500012 PM 12108501 ER PT J AU Scott, MC Wakamatsu, K Ito, S Kadekaro, AL Kobayashi, N Groden, J Kavanagh, R Takakuwa, T Virador, V Hearing, VJ Abdel-Malek, ZA AF Scott, MC Wakamatsu, K Ito, S Kadekaro, AL Kobayashi, N Groden, J Kavanagh, R Takakuwa, T Virador, V Hearing, VJ Abdel-Malek, ZA TI Human melanocortin 1 receptor variants, receptor function and melanocyte response to UV radiation SO JOURNAL OF CELL SCIENCE LA English DT Article DE melanocortin 1 receptor; human melanocytes; MC1R variants ID RED HAIR; MSH RECEPTOR; PIGMENTATION PHENOTYPE; GENE VARIANTS; SKIN-CANCER; FAIR SKIN; MELANIN; RISK; MC1R; PHEOMELANIN AB Cutaneous pigmentation is determined by the amounts of eumelanin and pheomelanin synthesized by epidermal melanocytes and is known to protect against sun-induced DNA damage. The synthesis of eumelanin is stimulated by the binding of alpha-melanotropin (alpha-melanocyte-stimulating hormone) to the functional melanocortin 1 receptor (MOR) expressed on melanocytes. The human MC1R gene is highly polymorphic and certain allelic variants of the gene are associated with red hair phenotype, melanoma and non-melanoma skin cancer. The importance of the MC1R gene in determining skin cancer risk led us to examine the impact of specific polymorphisms in this gene on the responses of human melanocytes to alpha-melanotropin and UV radiation. We compared the ability of human melanocyte cultures, each derived from a single donor, to respond to alpha-melanotropin with dose-dependent stimulation of cAMP formation, tyrosinase activity and proliferation. In each of those cultures the MC1R gene was sequenced, and the eumelanin and pheomelanin contents were determined. Human melanocytes homozygous; for Arg160Trp, heterozygous for Arg160Trp and Asp294His, or for Arg151Cys and Asp294His substitutions, but not melanocytes homozygous for Val92Met substitution, in the MC1R demonstrated a significantly reduced response to alpha-melanotropin. Additionally, melanocytes with a nonfunctional MC1R demonstrated a pronounced increase in their sensitivity to the cytotoxic effect of UV radiation compared with melanocytes expressing functional MC1R. We conclude that loss-of-function mutations in the MC1R gene sensitize human melanocytes to the DNA damaging effects of UV radiation, which may increase skin cancer risk. C1 Univ Cincinnati, Coll Med, Dept Dermatol, Cincinnati, OH 45267 USA. Fujita Hlth Univ, Sch Hlth Sci, Aichi 4701192, Japan. Nara Med Univ, Dept Dermatol, Nara 6348522, Japan. Univ Cincinnati, Coll Med, Dept Mol Genet Biochem & Microbiol, Cincinnati, OH 45267 USA. POLA Labs, Totsuka Ku, Yokohama, Kanagawa 2440812, Japan. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Abdel-Malek, ZA (reprint author), Univ Cincinnati, Coll Med, Dept Dermatol, POB 670592, Cincinnati, OH 45267 USA. FU NIEHS NIH HHS [R01 ES-09110] NR 45 TC 118 Z9 122 U1 0 U2 7 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD JUN 1 PY 2002 VL 115 IS 11 BP 2349 EP 2355 PG 7 WC Cell Biology SC Cell Biology GA 566WH UT WOS:000176450900010 PM 12006619 ER PT J AU Hoque, ATMS Yamano, S Liu, XB Swaim, WD Goldsmith, CM Delporte, C Baum, BJ AF Hoque, ATMS Yamano, S Liu, XB Swaim, WD Goldsmith, CM Delporte, C Baum, BJ TI Expression of the aquaporin 8 water channel in a rat salivary epithelial cell line SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Article ID FUNCTIONAL EXPRESSION; FLUID MOVEMENT; ACINAR-CELLS; IN-VITRO; MEMBRANE; TESTIS; IMMUNOLOCALIZATION; CLONING; GLANDS; LOCALIZATION AB Aquaporins are a family of water channels considered to play an important role in fluid transport across plasma membranes. Among the reported isoforms, relatively little is known about the functional role of aquaporin 8 (AQP8), and there are no cell lines known to express the AQP8 protein. We report here that the rat submandibular epithelial cell line, SMIE, expresses AQP8. Using RT-PCR, the presence of mRNA for AQP8 was demonstrated in these cells. Confocal immunofluorescence experiments revealed that the AQP8 protein is primarily present in the apical membranes of SMIE cells. When grown as a polarized monolayer on collagen coated polycarbonate filters, and exposed on their apical surface to different hyperosmotic (440, 540, or 640 mOsm) solutions, net fluid movement across SMIE cells was 8-25-fold that seen under isosmotic conditions. Similarly, when grown on coverslips and then exposed to a hypertonic solution, SMIE cells shrunk as a function of time, Together, these results suggest that SMIE cells endogenously express functional AQP8 water channels. Published 2002 Wiley-Liss, Inc.(dagger). C1 NIDCR, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Baum, BJ (reprint author), NIDCR, Gene Therapy & Therapeut Branch, NIH, Bldg 10,Rm 1N113,MSC-1190, Bethesda, MD 20892 USA. OI Yamano, Seiichi/0000-0003-2056-4359 NR 32 TC 12 Z9 14 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0021-9541 J9 J CELL PHYSIOL JI J. Cell. Physiol. PD JUN PY 2002 VL 191 IS 3 BP 336 EP 341 DI 10.1002/jcp.10106 PG 6 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 548XJ UT WOS:000175414900010 PM 12012329 ER PT J AU Alfaro, CL Wudarsky, M Nicolson, R Gochman, P Sporn, A Lenane, M Rapoport, JL AF Alfaro, CL Wudarsky, M Nicolson, R Gochman, P Sporn, A Lenane, M Rapoport, JL TI Correlation of antipsychotic and prolactin concentrations in children and adolescents acutely treated with haloperidol, clozapine, or olanzapine SO JOURNAL OF CHILD AND ADOLESCENT PSYCHOPHARMACOLOGY LA English DT Article ID CHILDHOOD-ONSET SCHIZOPHRENIA; SERUM PROLACTIN; CLINICAL-RESPONSE; HYPERPROLACTINEMIA; RISPERIDONE; DRUGS AB Patients with a Diagnostic and Statistical Manual of Mental Disorders (third edition, revised) diagnosis of schizophrenia or psychotic disorder not otherwise specified with onset of psychosis before the age of 13 participated in 6- to 8-week open or double-blind trials of haloperidol (n=15, mean dose 15.4+/-8.1 mg/day [0.27+/-0.15 mg/kg/day]), clozapine (n=30, mean dose 269.9+/-173.3 mg/day [4.4+/-2.6 mg/kg/day]), or olanzapine (n=12, mean dose 17.5+/-2.8 mg/day [0.30+/-0.13 mg/kg/day]). Blood samples were obtained at 6 weeks for evaluation of haloperidol, reduced haloperidol, clozapine, desmethylclozapine, and olanzapine plasma concentrations and serum prolactin concentrations. No gender differences were noted for antipsychotic dose or concentration within each treatment group. Correlations between antipsychotic plasma concentration and serum prolactin concentration were significant only for the olanzapine treatment group (r=0.80, p=0.002). Separate correlations for gender were significant only for females receiving olanzapine (r=0.91, p=0.03); the patient with the highest serum prolactin experienced galactorrhea. Further studies evaluating the prolactin-elevating properties of antipsychotics are warranted in this population. C1 NIH, Ctr Clin, Dept Pharm, Bethesda, MD 20892 USA. NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. RP Alfaro, CL (reprint author), NIH, Ctr Clin, Dept Pharm, Bldg 10,Room 1N257, Bethesda, MD 20892 USA. RI Nicolson, Robert/E-4797-2011 NR 28 TC 49 Z9 49 U1 2 U2 4 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1044-5463 J9 J CHILD ADOL PSYCHOP JI J. Child Adolesc. Psychopharmacol. PD SUM PY 2002 VL 12 IS 2 BP 83 EP 91 DI 10.1089/104454602760219126 PG 9 WC Pediatrics; Pharmacology & Pharmacy; Psychiatry SC Pediatrics; Pharmacology & Pharmacy; Psychiatry GA 576HM UT WOS:000176997900002 PM 12188977 ER PT J AU Romero, R Kuivaniemi, H Tromp, G AF Romero, R Kuivaniemi, H Tromp, G TI Editorial: Functional genomics and proteomics in term and preterm parturition SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Editorial Material ID HUMAN FETAL MEMBRANES; DIFFERENTIAL DISPLAY; GENE-EXPRESSION; AMNIOTIC-FLUID; LABOR; INTERLEUKIN-8; MYOMETRIUM; RUPTURE; MICROARRAYS; MECHANISMS C1 NICHHD, Perinatol Res Branch, Intramural Div, Detroit, MI 48201 USA. Wayne State Univ, Sch Med, Ctr Mol Med & Genet, Detroit, MI 48201 USA. Wayne State Univ, Sch Med, Dept Surg, Detroit, MI 48201 USA. RP Romero, R (reprint author), NICHHD, Perinatol Res Branch, Intramural Div, 4707 St Antoine Blvd, Detroit, MI 48201 USA. EM warfiela@mail.nih.gov RI Tromp, Gerard/B-2677-2017; OI Tromp, Gerard/0000-0002-7761-0806; Kuivaniemi, Helena/0000-0001-5753-8766 NR 46 TC 34 Z9 34 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 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD JUN PY 2002 VL 87 IS 6 BP 2431 EP 2434 DI 10.1210/jc.87.6.2431 PG 4 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 563DW UT WOS:000176241000003 PM 12050194 ER PT J AU Kalantaridou, SN Chrousos, GP AF Kalantaridou, SN Chrousos, GP TI Monogenic disorders of puberty SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Review ID FOLLICLE-STIMULATING-HORMONE; ADRENAL HYPOPLASIA CONGENITA; ACUTE REGULATORY PROTEIN; COMPOUND HETEROZYGOUS MUTATION; MCCUNE-ALBRIGHT-SYNDROME; RECEPTOR-GENE-MUTATIONS; LEYDIG-CELL HYPOPLASIA; STEROID 5-ALPHA-REDUCTASE-2 DEFICIENCY; COMPLETE HYPOGONADOTROPIC HYPOGONADISM; ANDROGEN INSENSITIVITY SYNDROME C1 Univ Ioannina, Sch Med, Dept Obstet & Gynecol, Div Reprod Endocrinol, Ioannina 45500, Greece. NICHHD, Pediat Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Kalantaridou, SN (reprint author), Univ Hosp Ioannina, Dept Obstet & Gynecol, Panepistimiou Ave, Ioannina 45500, Greece. EM kalantas@exchange.nih.gov NR 138 TC 26 Z9 33 U1 0 U2 2 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD JUN PY 2002 VL 87 IS 6 BP 2481 EP 2494 DI 10.1210/jc.87.6.2481 PG 14 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 563DW UT WOS:000176241000012 PM 12050203 ER PT J AU Vottero, A Kino, T Combe, H Lecomte, P Chrousos, GP AF Vottero, A Kino, T Combe, H Lecomte, P Chrousos, GP TI A novel, C-terminal dominant negative mutation of the GR causes familial glucocorticoid resistance through abnormal interactions with p160 steroid receptor coactivators SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID PRIMARY CORTISOL RESISTANCE; NUCLEAR RECEPTOR; ESTROGEN-RECEPTOR; BETA-ISOFORM; PROTEIN-1 GRIP1; BINDING DOMAINS; 26S PROTEASOME; EXPRESSION; LOCALIZATION; TRAFFICKING AB Primary cortisol resistance is a rare, inherited or sporadic form of generalized end-organ insensitivity to glucocorticoids. Here, we report a kindred in which affected members had a heterozygous T to G base substitution at nucleotide 2373 of exon 9alpha of the GR gene, causing substitution of Ile by Met at position 747. This mutation was located close to helix 12, at the C terminus of the ligand-binding domain, which has a pivotal role in the formation of activation function (AF)-2, a subdomain that interacts with p160 coactivators. The affinity of the mutant GR for dexamethasone was decreased by about 2-fold, and its transcriptional activity on the glucocorticoid-responsive mouse mammary tumor virus promoter was compromised by 20- to 30-fold. In addition, the mutant GR functioned as a dominant negative inhibitor of wild-type receptor-induced transactivation. The mutant GR through its intact AF-1 domain bound to a p160 coactivator, but failed to do so through its AF-2 domain. Overexpression of a p160 coactivator restored the transcriptional activity and reversed the negative transdominant activity of the mutant GR. Interestingly, green fluorescent protein (GFP)-fused GRalphaI747M had a slight delay in its translocation from the cytoplasm into the nucleus and formed coarser nuclear speckles than GFP-fused wildtype GRalpha. Similarly, a GFP-fused p160 coactivator had a distinctly different distribution in the nucleus in the presence of mutant vs. wild-type receptor, presenting also as coarser speckling. We conclude that the mutation at amino acid 747 of the GR causes familial, autosomal dominant glucocorticoid resistance by decreasing ligand binding affinity and transcriptional activity, and by exerting a negative transdominant effect on the wild-type receptor. The mutant receptor has an ineffective AF-2 domain, which leads to an abnormal interaction with p160 coactivators and a distinct nuclear distribution of both. C1 NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. CHU Tours, Endocrine Unit, F-37044 Tours, France. RP Kino, T (reprint author), NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bldg 10,Room 9D42,10 Ctr Dr,MSC 1583, Bethesda, MD 20892 USA. EM kinot@mail.nih.gov NR 38 TC 73 Z9 83 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD JUN PY 2002 VL 87 IS 6 BP 2658 EP 2667 DI 10.1210/jc.87.6.2658 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 563DW UT WOS:000176241000036 PM 12050230 ER PT J AU Uwaifo, GI Parikh, SJ Keil, M Elberg, J Chin, J Yanovski, JA AF Uwaifo, GI Parikh, SJ Keil, M Elberg, J Chin, J Yanovski, JA TI Comparison of insulin sensitivity, clearance, and secretion estimates using euglycemic and hyperglycemic clamps in children SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID C-PEPTIDE CONCENTRATIONS; MINIMAL-MODEL; GLUCOSE-TOLERANCE; WHITE AMERICANS; AFRICAN-AMERICAN; ADOLESCENTS; RESISTANCE; PROINSULIN; OBESITY; INVIVO AB The euglycemic clamp is the gold standard for estimating insulin sensitivity. The hyperglycemic clamp is easier to perform and is the gold standard for estimating beta-cell secretion. Reports in adults suggest that hyperglycemic clamps can estimate insulin sensitivity with results equivalent to euglycemic clamps. We investigated whether insulin sensitivity measures from both clamps are equivalent in children. Thirty-one lean and obese children (mean body mass index, 25.1 +/- 4.9 kg/m(2); mean age, 8.7 +/- 1.4 yr; 15 girls and 16 boys; 12 black and 19 white) were studied. All subjects underwent hyperglycemic clamps, then euglycemic clamps 2-6 wk later. Body composition was estimated by dual energy x-ray absorptiometry. Visceral and se abdominal fat was estimated by abdominal magnetic resonance imaging. Whole-body glucose disposal and insulin sensitivity (SI clamp) derived from both clamps and normalized for total or visceral fat and lean mass were significantly correlated (r, 0.45-0.65; P < 0.05). However, absolute SI clamp values were not equivalent. Bland-Altman comparisons found that SI clamp estimates from hyperglycemic clamps became less precise as SI clamp increased. There were significant correlations between indices of beta-cell secretion from the hyperglycemic clamp and mean C-peptide values from the euglycemic clamp (P < 0.05). However, no correlation was found between measures of total insulin clearance (derived from the euglycemic clamp) and surrogates of hepatic insulin clearance (derived from the hyperglycemic clamp). In this cohort of diverse children, SI clamp values from euglycemic and hyperglycemic clamps were significantly correlated but were not equivalent, whereas the insulin clearance measures were not correlated. It cannot be assumed that the hyperglycemic clamp obviates the need for euglycemic clamp studies to accurately estimate insulin sensitivity in children. C1 NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Uwaifo, GI (reprint author), NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, 10 Ctr Dr,Bldg 10,Room 10N262,MSC 1862, Bethesda, MD 20892 USA. EM Uwaifog@mail.nih.gov RI Uwaifo, Gabriel/M-2361-2016 OI Uwaifo, Gabriel/0000-0002-6962-9304 FU NICHD NIH HHS [HD-00641] NR 53 TC 27 Z9 28 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD JUN PY 2002 VL 87 IS 6 BP 2899 EP 2905 DI 10.1210/jc.87.6.2899 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 563DW UT WOS:000176241000071 PM 12050270 ER PT J AU Charmandari, E AF Charmandari, E TI Serum cortisol and 17-hydroxyprogesterone concentrations in children with classic congenital adrenal hyperplasia - Response SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Letter ID 21-HYDROXYLASE DEFICIENCY C1 NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Rockville, MD 20852 USA. RP Charmandari, E (reprint author), NICHHD, Pediat & Reprod Endocrinol Branch, NIH, 10 Ctr Dr,Bldg 10,Room 9D42, Rockville, MD 20852 USA. NR 6 TC 0 Z9 0 U1 1 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD JUN PY 2002 VL 87 IS 6 BP 2993 EP 2993 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 563DW UT WOS:000176241000090 ER PT J AU Wang, HY Fu, T Wang, G Zeng, G Perry-Lalley, DM Yang, JC Restifo, NP Hwu, P Wang, RF AF Wang, HY Fu, T Wang, G Zeng, G Perry-Lalley, DM Yang, JC Restifo, NP Hwu, P Wang, RF TI Induction of CD4(+) T cell-dependent antitumor immunity by TAT-mediated tumor antigen delivery into dendritic cells SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID TYROSINASE-RELATED PROTEIN-2; CLASS-I PRESENTATION; INTRACELLULAR DELIVERY; MELANOMA PATIENTS; B16 MELANOMA; HUMAN CANCER; PEPTIDE; VACCINATION; RESPONSES; VIVO AB Dendritic cell-based (DC-based) immunotherapy represents a promising approach to the prevention and treatment of many diseases, including cancer, but current strategies have met with only limited success in clinical and preclinical studies. Previous studies have demonstrated that a TAT peptide derived from the HIV TAT protein has the ability to transduce peptides or proteins into various cells. Here, we describe the use of TAT-mediated delivery of T cell peptides into DCs to prolong antigen presentation and enhance T cell responses. While immunization of mice with DCs pulsed with an antigenic peptide derived from the human TRP2 protein generated partial protective immunity against B16 tumor, immunization with DCs loaded with a TAT-TRP2 peptide resulted in complete protective immunity, as well as significant inhibition of lung metastases in a 3-day tumor model. Although both DC/TRP2 and DC/TAT-TRP2 immunization increased the number of TRP2-specific CD8(+) T cells detected by K-b/TRP2 tetramers, T cell activity elicited by DC/TAT-TRP2 was three-to tenfold higher than that induced by DC/TRP2. Furthermore, both CD4(+) and CD8(+) T cells were required for antitumor immunity demonstrated by experiments with antibody depletion of subsets of T cells, as well as with various knockout mice. These results suggest that a TAT-mediated antigen delivery system may have important clinical applications for cancer therapy. C1 Baylor Coll Med, Ctr Cell & Gene Therapy, Houston, TX 77030 USA. Baylor Coll Med, Dept Immunol, Houston, TX 77030 USA. NCI, Surg Branch, NIH, Bethesda, MD USA. RP Wang, RF (reprint author), Baylor Coll Med, Ctr Cell & Gene Therapy, 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 [Z01 BC010763-01, Z99 CA999999]; NCI NIH HHS [R01 CA090327, R01-CA90327-01A1] NR 50 TC 67 Z9 72 U1 0 U2 0 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD JUN PY 2002 VL 109 IS 11 BP 1463 EP 1470 DI 10.1172/JCI200215399 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 559FN UT WOS:000176015300010 PM 12045260 ER PT J AU Orange, JS Brodeur, SR Jain, A Bonilla, FA Schneider, LC Kretschmer, R Nurko, S Rasmussen, WL Kohler, JR Gellis, SE Ferguson, BM Strominger, JL Zonana, J Ramesh, N Ballas, ZK Geha, RS AF Orange, JS Brodeur, SR Jain, A Bonilla, FA Schneider, LC Kretschmer, R Nurko, S Rasmussen, WL Kohler, JR Gellis, SE Ferguson, BM Strominger, JL Zonana, J Ramesh, N Ballas, ZK Geha, RS TI Deficient natural killer cell cytotoxicity in patients with IKK-gamma/NEMO mutations SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID NF-KAPPA-B; HYPOHIDROTIC ECTODERMAL DYSPLASIA; MURINE CYTOMEGALOVIRUS-INFECTION; INCONTINENTIA PIGMENTI; NK CELLS; KINASE; ACTIVATION; NEMO; EXPRESSION; RECEPTORS AB NF-kappaB essential modifier (NEMO), also known as IKK-gamma, is a member of the I-kappaB kinase complex responsible for phosphorylating I-kappaB, allowing the release and activation of NF-kappaB. Boys with an expressed NEMO mutation have an X-linked syndrome characterized by hypohidrotic ectodermal dysplasia with immune deficiency (HED-ID). The immunophenotype resulting from NEMO mutation is highly variable, with deficits in both T and B cell responses. We evaluated three patients with NEMO mutations (L153, Q403X, and C417R) and HED-ID who had evidence of defective CD40 signaling. All three patients had normal percentages of peripheral blood NK cells, but impaired NK cell cytotoxic activity. This was not due to a generalized defect in cytotoxicity because antibody-dependent cellular cytotoxicity was intact. This abnormality was partially reversed by in vitro addition of IL-2 which was also able to induce NF-kappaB activation. In one patient with recurrent cytomegalovirus infections, administration of IL-2 partially corrected the NK cell killing deficit. These data suggest that NEMO participates in signaling pathways leading to NK cell cytotoxicity and that IL-2 can activate NF-kappaB and partially overcome the NK cell defect in patients with NEMO mutations. C1 Childrens Hosp, Div Immunol, Boston, MA 02115 USA. Harvard Univ, Sch Med, Dept Pediat, Boston, MA USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Hosp ABC, Dept Pediat, Mexico City, DF, Mexico. Iowa City Vet Adm, Dept Internal Med, Iowa City, IA USA. Harvard Univ, Sch Med, Dept Pediat, Boston, MA 02115 USA. Childrens Hosp, Div Infect Dis, Boston, MA 02115 USA. Oregon Hlth Sci Univ, Dept Mol & Med Genet, Portland, OR 97201 USA. Harvard Univ, Dept Cell & Mol Biol, Cambridge, MA 02138 USA. RP Geha, RS (reprint author), Childrens Hosp, Div Immunol, 300 Longwood Ave, Boston, MA 02115 USA. RI Orange, Jordan/D-5239-2009; OI orange, jordan/0000-0001-7117-7725 FU NCRR NIH HHS [M01 RR002172, M01-RR02172]; NIAID NIH HHS [AI-31136, P01 AI031541, R01 AI031136, AI-07512, AI-31541, U19 AI031541, T32 AI007512]; NIDCR NIH HHS [DE-11311, R01 DE011311] NR 39 TC 133 Z9 138 U1 0 U2 4 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD JUN PY 2002 VL 109 IS 11 BP 1501 EP 1509 DI 10.1172/JCI200214858 PG 9 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 559FN UT WOS:000176015300014 PM 12045264 ER PT J AU Yepes, M Sandkvist, M Coleman, TA Moore, E Wu, JY Mitola, D Bugge, TH Lawrence, DA AF Yepes, M Sandkvist, M Coleman, TA Moore, E Wu, JY Mitola, D Bugge, TH Lawrence, DA TI Regulation of seizure spreading by neuroserpin and tissue-type plasminogen activator is plasminogen-independent SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID INDUCED NEURONAL DEGENERATION; KAINIC ACID; STATUS EPILEPTICUS; MICE; HIPPOCAMPAL; INHIBITOR; GENE; INDUCTION; SUBSTRATE; DEMENTIA AB Tissue-type plasminogen activator (tPA) is a highly specific serine proteinase expressed in the CNS during events that require neuronal plasticity. In this study we demonstrate that endogenous tPA mediates the progression of kainic acid-induced (KA-induced) seizures by promoting the synchronization of neuronal activity required for seizure spreading, and that, unlike KA-induced cell death, this activity is plasminogen-independent. Specifically, seizure induction by KA injection into the amygdala induces tPA activity and cell death in both hippocampi, and unilateral treatment of rats with neuroserpin, a natural inhibitor of tPA in the brain, enhances neuronal survival in both hippocampi. Inhibition of tPA within the hippocampus by neuroserpin treatment does not prevent seizure onset but instead markedly delays the progression of seizure activity in both rats and wild-type mice. In tPA-deficient mice, seizure progression is significantly delayed, and neuroserpin treatment does not further delay seizure spreading. In contrast, plasminogen-deficient mice show a pattern of seizure spreading and a response to neuroserpin that is similar to that of wild-type animals. These findings indicate that tPA acts on a substrate other than plasminogen and that the effects of neuroserpin on seizure progression and neuronal cell survival are mediated through the inhibition of tPA. C1 Amer Red Cross, Holland Lab, Vasc Biol Dept, Rockville, MD 20855 USA. Georgetown Univ Hosp, Dept Neurol, Washington, DC 20007 USA. Amer Red Cross, Holland Lab, Dept Biochem, Rockville, MD 20855 USA. Human Genome Sci Inc, Dept Prot Dev, Rockville, MD USA. Georgetown Univ, Sch Med, Dept Physiol & Biophys, Washington, DC 20007 USA. NIDCR, Proteases & Tissue Remodeling Unit, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD USA. RP Lawrence, DA (reprint author), Amer Red Cross, Holland Lab, Vasc Biol Dept, 15601 Crabbs Branch Way, Rockville, MD 20855 USA. RI Yepes, Manuel/C-3576-2011 FU NHLBI NIH HHS [R01 HL055374, HL-55374, HL-55747, R01 HL055747]; NINDS NIH HHS [K08 NS002223, NS-02223, NS-36477] NR 34 TC 93 Z9 98 U1 0 U2 3 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD JUN PY 2002 VL 109 IS 12 BP 1571 EP 1578 DI 10.1172/JCI200214308 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 564MR UT WOS:000176318600010 PM 12070304 ER PT J AU Yang, YA Dukhanina, O Tang, BW Mamura, M Letterio, JJ MacGregor, J Patel, SC Khozin, S Liu, ZY Green, J Anver, MR Merlino, G Wakefield, LM AF Yang, YA Dukhanina, O Tang, BW Mamura, M Letterio, JJ MacGregor, J Patel, SC Khozin, S Liu, ZY Green, J Anver, MR Merlino, G Wakefield, LM TI Lifetime exposure to a soluble TGF-beta antagonist protects mice against metastasis without adverse side effects SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID GROWTH-FACTOR-BETA; TRANSGENIC MICE; BREAST-CANCER; II RECEPTOR; TRANSFORMING GROWTH-FACTOR-BETA-1; EXPERIMENTAL GLOMERULONEPHRITIS; MAMMARY-GLAND; IN-VIVO; T-CELLS; DISEASE AB TGF-betas play diverse and complex roles in many biological processes. In tumorigenesis, they can function either as tumor suppressors or as pro-oncogenic factors, depending on the stage of the disease. We have developed transgenic mice expressing a TGF-beta antagonist of the soluble type II TGF-beta receptor:Fc fusion protein class, under the regulation of the mammary-selective MMTV-LTR promoter/enhancer. Biologically significant levels of antagonist were detectable in the serum and most tissues of this mouse line. The mice were resistant to the development of metastases at multiple organ sites when compared with wild-type controls, both in a tail vein metastasis assay using isogenic melanoma cells and in crosses with the MMTV-neu transgenic mouse model of metastatic breast cancer. Importantly, metastasis from endogenous mammary tumors was suppressed without any enhancement of primary tumorigenesis. Furthermore, aged transgenic mice did not exhibit the severe pathology characteristic of TGF-beta null mice, despite lifetime exposure to the antagonist. The data suggest that in vivo the antagonist may selectively neutralize the undesirable TGF-beta associated with metastasis, while sparing the regulatory roles of TGF-betas in normal tissues. Thus this soluble TGF-beta antagonist has potential for long-term clinical use in the prevention of metastasis. C1 NCI, Lab Cell Regulat & Carcinogenesis, Bethesda, MD 20892 USA. NCI, Pathol & Histotechnol Lab, Sci Applicat Int Corp, Frederick, MD 21701 USA. NCI, Mol Biol Lab, Bethesda, MD 20892 USA. RP Wakefield, LM (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, Bldg 41,Rm C629,41 Lib Dr,MSC 5055, Bethesda, MD 20892 USA. OI Mamura, Mizuko/0000-0003-4531-0144 FU NCI NIH HHS [N01CO12400, N01-CO-12400] NR 47 TC 281 Z9 317 U1 0 U2 3 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA 35 RESEARCH DR, STE 300, ANN ARBOR, MI 48103 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD JUN PY 2002 VL 109 IS 12 BP 1607 EP 1615 DI 10.1172/JCI200215333 PG 9 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 564MR UT WOS:000176318600014 PM 12070308 ER PT J AU Dierker, LC Merikangas, KR AF Dierker, LC Merikangas, KR TI Design and interpretability of findings in a family study to investigate posttraumatic stress disorder - Drs. Dierker and Merikangas reply SO JOURNAL OF CLINICAL PSYCHIATRY LA English DT Letter ID INTERVIEW C1 Wesleyan Univ, Middletown, CT 06457 USA. NIMH, Washington, DC USA. RP Dierker, LC (reprint author), Wesleyan Univ, Middletown, CT 06457 USA. NR 3 TC 0 Z9 0 U1 0 U2 0 PU PHYSICIANS POSTGRADUATE PRESS PI MEMPHIS PA P O BOX 240008, MEMPHIS, TN 38124 USA SN 0160-6689 J9 J CLIN PSYCHIAT JI J. Clin. Psychiatry PD JUN PY 2002 VL 63 IS 6 BP 538 EP 538 PG 1 WC Psychology, Clinical; Psychiatry SC Psychology; Psychiatry GA 566GZ UT WOS:000176419700018 ER PT J AU Lerman, C Croyle, RT Tercyak, KP Hamann, H AF Lerman, C Croyle, RT Tercyak, KP Hamann, H TI Genetic testing: Psychological aspects and implications SO JOURNAL OF CONSULTING AND CLINICAL PSYCHOLOGY LA English DT Review ID BREAST-CANCER SUSCEPTIBILITY; SMOKING-CESSATION TREATMENT; AT-RISK RELATIVES; HUNTINGTON DISEASE; CYSTIC-FIBROSIS; OVARIAN-CANCER; COLON-CANCER; DECISION-MAKING; PRETEST EDUCATION; GENERAL-POPULATION AB As the number of genes associated with inherited disease continues to grow, researchers and practitioners in behavioral medicine will encounter complex psychological issues faced by individuals at risk for these diseases. A review of the literature concerning prenatal, carrier, and predictive genetic testing suggests that the severity of psychological risks posed by research-based genetic testing is not great. However, subgroups of individuals with particular psychological traits may be more vulnerable to adverse effects. Available data do not provide evidence that genetic testing promotes changes in health-related behaviors. Thus, although there may be less of a role for mental health professionals in the psychological counseling of genetic testing participants, there is a need for research and practice to facilitate health protective behaviors in response to genetic risk information. C1 Univ Penn, Ctr Canc, Philadelphia, PA 19104 USA. NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Georgetown Univ, Med Ctr, Lombardi Canc Ctr, Washington, DC 20057 USA. Univ Utah, Dept Psychol, Salt Lake City, UT 84112 USA. RP Lerman, C (reprint author), Univ Penn, Ctr Canc, 3535 Market St,Suite 4100, Philadelphia, PA 19104 USA. FU NCI NIH HHS [CA/HG74861]; NHGRI NIH HHS [HG01846] NR 128 TC 162 Z9 162 U1 4 U2 17 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0022-006X J9 J CONSULT CLIN PSYCH JI J. Consult. Clin. Psychol. PD JUN PY 2002 VL 70 IS 3 BP 784 EP 797 DI 10.1037//0022-006X.70.3.784 PG 14 WC Psychology, Clinical SC Psychology GA 562LJ UT WOS:000176198000022 PM 12090383 ER PT J AU Siegler, IC Bastian, LA Steffens, DC Bosworth, HB Costa, PT AF Siegler, IC Bastian, LA Steffens, DC Bosworth, HB Costa, PT TI Behavioral medicine and aging SO JOURNAL OF CONSULTING AND CLINICAL PSYCHOLOGY LA English DT Review ID PHYSICAL-ACTIVITY; LIFE-SPAN; ALZHEIMERS-DISEASE; COLLEGE ALUMNI; UNITED-STATES; RISK-FACTORS; AGE; PERSONALITY; HEALTH; POPULATION AB A brief history of behavioral medicine and aging is followed by a series of perspectives that help to understand how age is used as a variable in this research, the relative importance of age to declines in cognitive functioning, and the impact of behavioral risk indicators on healthy survival. The authors discuss Alzheimer's disease and the role of age in clinical practice. Also discussed are potential problems in age-related research, ways to improve the knowledge base in behavioral medicine and aging, and thoughts about future challenges to enhance work in behavioral medicine and aging. C1 Duke Univ, Med Ctr, Behav Med Res Ctr, Dept Psychiat & Behav Sci,Sch Med, Durham, NC 27710 USA. Duke Univ, Sch Med, Dept Med, Durham, NC 27710 USA. Duke Univ, Sch Med, Dept Obstet & Gynecol, Durham, NC 27710 USA. NIA, Lab Personal & Cognit, Bethesda, MD 20892 USA. RP Siegler, IC (reprint author), Duke Univ, Med Ctr, Behav Med Res Ctr, Dept Psychiat & Behav Sci,Sch Med, Box 2969, Durham, NC 27710 USA. EM ilene.siegler@duke.edu OI Costa, Paul/0000-0003-4375-1712 FU NCI NIH HHS [P01 CA72099]; NHLBI NIH HHS [P01 HL36587, R01HL55356]; NIA NIH HHS [R01 AG12458]; NIMH NIH HHS [K07 MH 01367] NR 103 TC 11 Z9 11 U1 1 U2 2 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0022-006X J9 J CONSULT CLIN PSYCH JI J. Consult. Clin. Psychol. PD JUN PY 2002 VL 70 IS 3 BP 843 EP 851 DI 10.1037//0022-006X.70.3.843 PG 9 WC Psychology, Clinical SC Psychology GA 562LJ UT WOS:000176198000026 PM 12090387 ER PT J AU Keller, RA Lipkus, IM Rimer, BK AF Keller, RA Lipkus, IM Rimer, BK TI Depressive realism and health risk accuracy: The negative consequences of positive mood SO JOURNAL OF CONSUMER RESEARCH LA English DT Article ID PROTECTION MOTIVATION THEORY; COMMUNITY SAMPLE; SELF-ENHANCEMENT; JUDGMENTS; PERSPECTIVE; PERSONALITY; ILLUSION; BEHAVIORS; ACCESSIBILITY; HELPLESSNESS AB We examine the role of level of depression on updating of health-related risk estimates. Participants provided their risk of getting breast cancer before (baseline) and after (follow-up) receiving personalized (experiment 1) or standard (experiment 2) medical risk feedback. Although there were no significant differences in risk estimates at baseline, the follow-up risk estimates indicate that compared to nondepressives, depressives lowered their risk estimates such that they were more accurate or closer to the medical estimates provided in the risk feedback. In contrast to depressives, nondepressives with higher baseline risk estimates did not revise their follow-up risk estimates because they were in a positive mood after receiving the risk feedback. C1 Dartmouth Coll, Tuck Sch Business, Hanover, NH 03755 USA. Duke Univ, Med Ctr, Durham, NC 27708 USA. NCI, Bethesda, MD 20892 USA. RP Keller, RA (reprint author), Dartmouth Coll, Tuck Sch Business, Hanover, NH 03755 USA. NR 77 TC 7 Z9 7 U1 4 U2 16 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0093-5301 J9 J CONSUM RES JI J. Consum. Res. PD JUN PY 2002 VL 29 IS 1 BP 57 EP 69 PG 13 WC Business SC Business & Economics GA 556NN UT WOS:000175856000004 ER PT J AU O'Grady, NP AF O'Grady, NP TI Applying the science to the prevention of catheter-related infections SO JOURNAL OF CRITICAL CARE LA English DT Article ID CENTRAL VENOUS CATHETER; BLOOD-STREAM INFECTION; INTENSIVE-CARE UNIT; RANDOMIZED CONTROLLED TRIAL; DOUBLE-BLIND TRIAL; SCANNING ELECTRON-MICROSCOPY; CRITICALLY ILL PATIENTS; STAPHYLOCOCCUS-AUREUS; MUPIROCIN OINTMENT; POVIDONE-IODINE AB Background. Catheter-related bloodstream infection (CR-BSI) remains a leading cause of nososcomial infection, despite the fact that many CR-BSIs are preventable. Simple principles of infection control and the use of novel devices to reduce these infections are not uniformly implemented. Objective:To review and summarize the evidence that promotes strategies to prevent CR-BSIs. Data sources: The Medline database and bibliographies of review articles and book chapters were searched for relevant articles. Studies included. In vitro and in vivo laboratory-based studies, controlled clinical trials, prospective interventional trials, and epidemiologic investigations. Outcome measures: Reduction in CR-BSI, catheter colonization, tunnel infection, or local site infection. Synthesis: The recommended prevention strategies with strong supportive evidence include educating and training of health care providers who insert and maintain catheters; using full barrier precautions during central venous catheter insertion; using a 2% chlorhexidine preparation for skin antisepsis; eliminating the practice of scheduled replacement of central venous catheters for prevention of infection; and using antiseptic/antibiotic impregnated short-term central venous catheters. Conclusion. Simple interventions can reduce the risk for serious catheter-related infection. Health care provider awareness and adherence to these prevention strategies is critical to reducing the risk for CR-BSI, improving patient safety, and promoting quality health care. Copyright 2002, Elsevier Science (USA). All rights reserved. C1 NIH, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP O'Grady, NP (reprint author), NIH, Warren G Magnuson Clin Ctr, 10 Ctr Dr,Bldg 10,Room 7D 43, Bethesda, MD 20892 USA. NR 67 TC 11 Z9 11 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 0883-9441 J9 J CRIT CARE JI J. Crit. Care PD JUN PY 2002 VL 17 IS 2 BP 114 EP 121 DI 10.1053/jcrc.2002.34366 PG 8 WC Critical Care Medicine SC General & Internal Medicine GA 568PC UT WOS:000176550100007 PM 12096374 ER PT J AU Nilsson, O Abad, V Chrysis, D Ritzen, EM Savendahl, L Baron, J AF Nilsson, O Abad, V Chrysis, D Ritzen, EM Savendahl, L Baron, J TI Estrogen receptor-alpha and -beta are expressed throughout postnatal development in the rat and rabbit growth plate SO JOURNAL OF ENDOCRINOLOGY LA English DT Article ID BONE-GROWTH; CARTILAGE CELLS; SKELETAL GROWTH; SEX STEROIDS; LOCALIZATION; ESTRADIOL; MICE; AGE; OSSIFICATION; CHONDROCYTES AB Estrogen regulates skeletal growth and promotes epiphyseal fusion. To explore the mechanisms underlying these effects we investigated the expression of estrogen receptor-alpha (ERalpha) and -beta (ERbeta) in rat and rabbit growth plates during postnatal development, using immunolustochemistry. Immunoreactiviry for ERalpha and ERbeta was observed in resting zone and proliferative zone chondrocytes at all ages studied for both rat (7, 14, 28 and 70 days of age) and rabbit (1, 7, 28 and 120 days of age). In the rat distal humerus and the rabbit proximal tibia, expression of both receptors in the hypertrophic zone was minimal at early ages, increasing only at the last time point prior to epiphyseal fusion. Expression was rarely seen in the hypertrophic zone of the rat proximal tibia, a growth plate that does not fuse until late in life. Therefore, we conclude that ERalpha and ERbeta are both expressed in the mammalian growth plate. The temporal and anatomical pattern suggests that ER expression in the hypertrophic zone in particular may play a role in epiphyseal fusion. C1 NICHHD, NIH, Dev Endocrinol Branch, Bethesda, MD 20892 USA. Karolinska Inst, Pediat Endocrinol Unit, Dept Woman & Child Hlth, Stockholm, Sweden. RP Nilsson, O (reprint author), NICHHD, NIH, Dev Endocrinol Branch, Bldg 10,Room 10N262,10 Ctr Dr,MSC 1862, Bethesda, MD 20892 USA. NR 39 TC 40 Z9 44 U1 2 U2 2 PU SOC ENDOCRINOLOGY PI BRISTOL PA 17/18 THE COURTYARD, WOODLANDS, BRADLEY STOKE, BRISTOL BS32 4NQ, ENGLAND SN 0022-0795 J9 J ENDOCRINOL JI J. Endocrinol. PD JUN PY 2002 VL 173 IS 3 AR UNSP 0022-0795/02/0173-407 DI 10.1677/joe.0.1730407 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 568LB UT WOS:000176542300003 PM 12065230 ER PT J AU Hackos, DH Chang, TH Swartz, KJ AF Hackos, DH Chang, TH Swartz, KJ TI Scanning the intracellular S6 activation gate in the shaker K+ channel SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Article DE Kv channel; voltage-dependent gating; scanning mutagenesis; pore occlusion; closed gate ID POTASSIUM CHANNELS; OPEN-STATE; INACTIVATION; TRANSITIONS; PORE; PROPENSITY; CONDUCTION; INHIBITORS; RECEPTOR; RESIDUES AB In Kv channels, an activation gate is thought to be located near the intracellular entrance to the ion conduction pore. Although the COOH terminus of the S6 segment has been implicated in forming the gate structure, the residues positioned at the occluding part of the gate remain undetermined. We use a mutagenic scanning approach in the Shaker Kv channel, mutating each residue in the S6 gate region (T469-Y485) to alanine, tryptophan, and aspartate to identify positions that are insensitive to mutation and to find mutants that disrupt the gate. Most mutants open in a steeply voltage-dependent manner and close effectively at negative voltages, indicating that the gate structure can both support ion flux when open and prevent it when closed. We find several mutant channels where macroscopic ionic currents are either very small or undetectable, and one mutant that displays constitutive currents at negative voltages. Collective examination of the three types of substitutions support the notion that the intracellular portion of S6 forms an activation gate and identifies V478 and F481 as candidates for occlusion of the pore in the closed state. C1 NINCDS, Mol Physiol & Biophys Unit, NIH, Bethesda, MD 20892 USA. RP Swartz, KJ (reprint author), NINCDS, Mol Physiol & Biophys Unit, NIH, Bldg 36,Rm 2C19,36 Convent Dr,MSC 4066, Bethesda, MD 20892 USA. FU Intramural NIH HHS [ZIA NS002945-13] NR 42 TC 128 Z9 130 U1 0 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0022-1295 J9 J GEN PHYSIOL JI J. Gen. Physiol. PD JUN PY 2002 VL 119 IS 6 BP 521 EP 531 DI 10.1085/jgp.20028569 PG 11 WC Physiology SC Physiology GA 562CV UT WOS:000176180600002 PM 12034760 ER PT J AU Fridman, AI Matveev, SA Agalakova, NI Fedorova, OV Lakatta, EG Bagrov, AY AF Fridman, AI Matveev, SA Agalakova, NI Fedorova, OV Lakatta, EG Bagrov, AY TI Marinobufagenin, an endogenous ligand of alpha-1 sodium pump, is a marker of congestive heart failure severity SO JOURNAL OF HYPERTENSION LA English DT Article DE bufanolides; heart failure; Na(+)-K(+)-exchanging ATPase; natriuretic peptides; ouabain; steroids ID NATRIURETIC-PEPTIDE SECRETION; OUABAIN-LIKE IMMUNOREACTIVITY; ESSENTIAL-HYPERTENSION; NA+/K+-ATPASE; PROTEIN-KINASE; CYCLIC-GMP; RAT AORTA; DIGITALIS; PLASMA; INHIBITION AB Background A reduced cardiac output in chronic heart failure (CHF) evokes renal NaCl and water retention, and, therefore, activates mechanisms promoting natriuresis. Atrial natriuretic peptide (ANP) is one such factor. We hypothesized that another NaCl sensitive endogenous natriuretic factor, i.e., marinobufagenin (MBG), a specific ligand of the alpha-1 subunit of Na/K ATPase (the main kidney isoform) and also a vasoconstrictor and cardiotonic substance, would be elevated in CHF patients in a graded manner with the severity of CHF. Methods and results We measured the plasma levels of MBG, alpha-hANP, ouabain-like compound (OLC) and left ventricular (LV) volumes and ejection fraction in 23 consecutive hypertensive male patients with CHF. Plasma MBG levels exhibited progressive increases (0.59 +/- 0.15, 1.08 +/- 0.20,1.35 +/- 0.17 and 1.88 +/- 0.05 nmol/l NYHA 1-4, respectively) and paralleled the changes of cc-hANP. Conversely, plasma OLC did not exhibit such increases. Plasma MBG correlated with alpha-hANP (r = 0.82; P< 0.0001). Both MBG and alpha-hANP correlated with LV systolic (r = 0.55 and r = 0.47; P< 0.01) diameter and inversely with ejection fraction (r = -0.73 and r = -0.60; P< 0.01). OLC did not exhibit correlations with alpha-hANP or LV volumes, but positively correlated with systolic brachial blood pressure and with pulse pressure. Conclusions In CHF, MBG exhibits progressive increases similar to ANP, varies with CHF severity and correlates with LV systolic function. We hypothesize, that, in CHF, the concurrent production of these two natriuretic hormones, a vasorelaxant, ANP, and a vasoconstrictor, MBG, potentiate each other's natriuretic effects, but may offset their vasoactive actions. (C) 2002 Lippincott Williams Wilkins. C1 NIA, Cardiovasc Sci Lab, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Mil Med Acad, Dept Cardiac Surg, St Petersburg, Russia. RP Bagrov, AY (reprint author), NIA, Cardiovasc Sci Lab, Intramural Res Program, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 50 TC 44 Z9 45 U1 0 U2 1 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 JUN PY 2002 VL 20 IS 6 BP 1189 EP 1194 DI 10.1097/00004872-200206000-00032 PG 6 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 559LL UT WOS:000176027600032 PM 12023690 ER PT J AU Kato, M Kawai, Y Yoshida, M Masamura, K Inazu, T Hayashi, T Miyamori, I AF Kato, M Kawai, Y Yoshida, M Masamura, K Inazu, T Hayashi, T Miyamori, I TI Amphiregulin stimulates vascular smooth muscle cells (VSMC) proliferation in an autocrine and/or paracrine manner via-MAPK and PI3K-Akt/PBK pathway SO JOURNAL OF HYPERTENSION LA English DT Meeting Abstract CT 19th Meeting of the International-Society-for-Hypertension/12th European-Society-of-Hypertension Meeting CY JUN 23-27, 2002 CL PRAGUE, CZECH REPUBLIC SP Int Soc Hypertens, European Soc Hypertens C1 Fukui Med Univ, Dept Internal Med 3, Fukui, Japan. NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 1 U2 1 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 JUN PY 2002 VL 20 SU 4 MA P83 BP S31 EP S31 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 600JF UT WOS:000178386300136 ER PT J AU Pacak, K Carrasquillo, JA Chen, CA Eisenhofer, G Li, S Goldstein, DS AF Pacak, K Carrasquillo, JA Chen, CA Eisenhofer, G Li, S Goldstein, DS TI Pheochromocytoma: Novel approach to localization by 6-[F-18]fluorodopamine positron emission tomographic (PET) scanning SO JOURNAL OF HYPERTENSION LA English DT Meeting Abstract CT 19th Meeting of the International-Society-for-Hypertension/12th European-Society-of-Hypertension Meeting CY JUN 23-27, 2002 CL PRAGUE, CZECH REPUBLIC SP Int Soc Hypertens, European Soc Hypertens C1 NICHD, Pediat & Reprod Endocrinol Branch, Bethesda, MD USA. NIH, Dept Nucl Med, Ctr Clin, Bethesda, MD 20892 USA. RI Carrasquillo, Jorge/E-7120-2010 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 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD JUN PY 2002 VL 20 SU 4 MA O96 BP S172 EP S172 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 600JF UT WOS:000178386300796 ER PT J AU Scuteri, A Manolio, TA Lakatta, EG AF Scuteri, A Manolio, TA Lakatta, EG TI Specific carotid geometric phenotypes predict incidence of CV events in older persons. The cardiovascular health study SO JOURNAL OF HYPERTENSION LA English DT Meeting Abstract CT 19th Meeting of the International-Society-for-Hypertension/12th European-Society-of-Hypertension Meeting CY JUN 23-27, 2002 CL PRAGUE, CZECH REPUBLIC SP Int Soc Hypertens, European Soc Hypertens C1 NIA, NIH, Baltimore, MD 21224 USA. NHLBI, NIH, Bethesda, MD 20892 USA. INRCA, Rome, Italy. 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 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD JUN PY 2002 VL 20 SU 4 MA O66 BP S90 EP S90 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 600JF UT WOS:000178386300415 ER PT J AU Scuteri, A Muller, D Aandres, R Hogaku, H Metter, JE Lakatta, EG AF Scuteri, A Muller, D Aandres, R Hogaku, H Metter, JE Lakatta, EG TI Metabolic syndrome accelerates vascular aging SO JOURNAL OF HYPERTENSION LA English DT Meeting Abstract CT 19th Meeting of the International-Society-for-Hypertension/12th European-Society-of-Hypertension Meeting CY JUN 23-27, 2002 CL PRAGUE, CZECH REPUBLIC SP Int Soc Hypertens, European Soc Hypertens C1 NIA, NIH, Baltimore, MD 21224 USA. INRCA, Rome, Italy. 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 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD JUN PY 2002 VL 20 SU 4 MA P234 BP S62 EP S62 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 600JF UT WOS:000178386300284 ER PT J AU Sharabi, Y Grotto, I Heruti, R Huerta, M Grossman, E AF Sharabi, Y Grotto, I Heruti, R Huerta, M Grossman, E TI The association between socioeconomic parameters and blood pressure: Results of a large-scale population study - The Young Adult Periodic Examination in Israel (YAPEIS) database SO JOURNAL OF HYPERTENSION LA English DT Meeting Abstract CT 19th Meeting of the International-Society-for-Hypertension/12th European-Society-of-Hypertension Meeting CY JUN 23-27, 2002 CL PRAGUE, CZECH REPUBLIC SP Int Soc Hypertens, European Soc Hypertens C1 NINDS, NIH, Bethesda, MD 20892 USA. Period Examinat Ctr, Tel Hashomer, Israel. Chaim Sheba Med Ctr, Dept Internal Med D, IL-52621 Tel Hashomer, Israel. RI Grotto, Itamar/F-2028-2012 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 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD JUN PY 2002 VL 20 SU 4 MA P1399 BP S328 EP S328 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 600JF UT WOS:000178386301511 ER PT J AU Basta, S Chen, WS Bennink, JR Yewdell, JW AF Basta, S Chen, WS Bennink, JR Yewdell, JW TI Inhibitory effects of cytomegalovirus proteins US2 and US11 point to contributions from direct priming and cross-priming in induction of vaccinia virus-specific CD8(+) T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MHC CLASS-I; ANTIGEN PRESENTATION; EXOGENOUS ANTIGEN; DENDRITIC CELLS; DEFICIENT MICE; CUTTING EDGE; EXPRESSION; RESPONSES; IMMUNODOMINANCE; ADENOVIRUSES AB The extent to which naive CD8(+) CTLs (T-CD8(+)) are primed by APCs presenting endogenous Ags (direct priming) or Ags acquired from other infected cells (cross-priming) is a critical topic in basic and applied immunology. To examine the contribution of direct priming in the induction of NW-specific T-CD8(+), we generated recombinant vaccinia viruses that express human CMV proteins (US2 and US11) that induce the destruction of newly synthesized MHC class I molecules. Expression of US2 or US11 was associated with a 24-63% decrease in numbers of primary or secondary VV-specific T-CD8(+) responding to i.p. infection. Using HPLC-isolated peptides from NW-infected cells, we show that US2 and US11 selectively inhibit T-CD8(+) responses to a subset of immunogenic VV determinants. Moreover, VV-US2 and lysates from VV-infected histoincompatible cells elicit T-CD8(+) specific for a similar subset of VV determinants. These findings indicate that US2 and US11 can function in vivo to interfere with the activation of virus-specific T-CD8(+). Furthermore, they suggest that 1) both cross-priming and direct priming contribute significantly to the generation of VV-specific T-CD8(+), 2) the sets of immunogenic vaccinia virus determinants generated by cross-priming and direct priming are not completely overlapping, and 3) cross-priming overrides the effects of cis-acting viral interference with the class I Ag presentation pathway. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. Ludwig Inst Canc Res, Canc Vaccine Unit, Heidelberg, Germany. RP Yewdell, JW (reprint author), NIAID, Viral Dis Lab, NIH, Room 211,Bldg 4,4 Ctr Dr, Bethesda, MD 20892 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012; Chen, Weisan/E-7828-2012 NR 39 TC 47 Z9 47 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 JUN 1 PY 2002 VL 168 IS 11 BP 5403 EP 5408 PG 6 WC Immunology SC Immunology GA 555WA UT WOS:000175815200004 PM 12023332 ER PT J AU Roberts, TJ Sriram, V Spence, PM Gui, M Hayakawa, K Bacik, I Bennink, JR Yewdell, JW Brutkiewicz, RR AF Roberts, TJ Sriram, V Spence, PM Gui, M Hayakawa, K Bacik, I Bennink, JR Yewdell, JW Brutkiewicz, RR TI Recycling CD1d1 molecules present endogenous antigens processed in an endocytic compartment to NKT cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INNATE IMMUNE-SYSTEM; T-CELLS; LIPOGLYCAN ANTIGENS; CUTTING EDGE; ASIALOGLYCOPROTEIN RECEPTOR; STRUCTURAL REQUIREMENTS; LYSOSOMOTROPIC AMINES; CULTURED-CELLS; HEPATOMA-CELLS; MHC MOLECULES AB Mouse CD1d1 molecules present endogenous glycolipids to NKT cells. Although glycolipid presentation requires CD1d1 transport through the endocytic pathway, the processing requirements for such endogenous Ag presentation by CD1d1 molecules are undefined. We examined CD1d1 Ag presentation to NKT cells by disrupting endocytic trafficking and function in cells expressing normal and mutated CD1d1 expressed by recombinant vaccinia viruses. Consistent with previous studies, we found that preventing CD1d1 localization to endosomes by altering its cytoplasmic targeting sequences abrogated recognition by Valpha14Jalpha281(+) NKT cells without affecting recognition by Valpha14(-) NKT cells. Increasing the pH of acidic compartments by incubating cells with chloroquine or bafilomycin A1 blocked CD1d1 recognition by Valpha14(+) (but not Valpha14(-)) NKT cells without reducing levels of cell surface CD1d1. Similar results were obtained with primaquine, which interferes with the recycling of cell surface glycoproteins. These results suggest that the loading of a subset of glycolipid ligands onto CD1d1 molecules entails the delivery of cell surface CD1d1 molecules and an acidic environment in the endocytic pathway. C1 Indiana Univ, Sch Med, Dept Microbiol & Immunol, Indianapolis, IN 46202 USA. Walther Oncol Ctr, Indianapolis, IN 46202 USA. Walther Canc Inst, Indianapolis, IN 46208 USA. Fox Chase Canc Ctr, Inst Canc Res, Philadelphia, PA 19111 USA. NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Brutkiewicz, RR (reprint author), Indiana Univ, Sch Med, Dept Microbiol & Immunol, Bldg R4,Room 302,1044 W Walnut St, Indianapolis, IN 46202 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012; OI Webb, Tonya/0000-0003-3086-6414 FU NIAID NIH HHS [R01 AI 41726, AI46455-01S1, R01 AI 46455] NR 58 TC 89 Z9 92 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 JUN 1 PY 2002 VL 168 IS 11 BP 5409 EP 5414 PG 6 WC Immunology SC Immunology GA 555WA UT WOS:000175815200005 PM 12023333 ER PT J AU Sehgal, D Obiakor, H Mage, RG AF Sehgal, D Obiakor, H Mage, RG TI Distinct clonal Ig diversification patterns in young appendix compared to antigen-specific splenic clones SO JOURNAL OF IMMUNOLOGY LA English DT Article ID DOUBLE-STRAND BREAKS; GENE-CONVERSION-LIKE; B-CELL DEVELOPMENT; V-H GENES; SOMATIC HYPERMUTATION; GERMINAL-CENTERS; HEAVY-CHAIN; ANTIBODY REPERTOIRE; IMMUNOGLOBULIN GENES; REGION SEQUENCES AB The young rabbit appendix is a dynamic site for primary B cell repertoire development. To study diversification patterns during clonal expansion, we collected single appendix B cells from 3- to 9-wk-old rabbits and sequenced rearranged H and L chain genes. Single cells obtained by hydraulic micromanipulation or laser capture microdissection were lysed, PCR amplified, and products directly sequenced. Gene conversion-like changes occurred in rearranged H and L chain sequences by 3-4 wk of age. Somatic mutations were found in the D regions that lack known conversion donors and probably also occurred in the V genes. A few small sets of clonally related appendix B cells were found at 3-5 wk; by 5.5 wk, some larger clones were recovered. The diversification patterns in the clones from appendix were strikingly different from those found previously in splenic germinal centers where an immunizing Ag was driving the expansion and selection process toward high affinity. Clonally related appendix B cells developed different amino acid sequences in each complementarity-determining region (CDR) including CDR3, whereas dominant clones from spleen underwent few changes in CDR3. The variety of combining sites generated by diversification within individual clones suggests that at least some clonal expansion and selection, known to require normal gut flora, may be driven through indirect effects of microbial components rather than solely by their recognition as specific foreign Ags. This diversity of combining sites within B cell clones supports the proposed role of appendix in generating the preimmune repertoire. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Mage, RG (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Room 11N311,10 Ctr Dr,MSC 1892, Bethesda, MD 20892 USA. NR 35 TC 24 Z9 25 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 JUN 1 PY 2002 VL 168 IS 11 BP 5424 EP 5433 PG 10 WC Immunology SC Immunology GA 555WA UT WOS:000175815200007 PM 12023335 ER PT J AU Livingston, B Crimi, C Newman, M Higashimoto, Y Appella, E Sidney, J Sette, A AF Livingston, B Crimi, C Newman, M Higashimoto, Y Appella, E Sidney, J Sette, A TI A rational strategy to design multiepitope immunogens based on multiple th lymphocyte epitopes SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HLA-DR; HELPER EPITOPES; CELL EPITOPES; PEPTIDE; ANTIGEN; VACCINE; BINDING; IDENTIFICATION; IMMUNIZATION; PROTEIN AB Four HLA-DR-restricted HIV-derived Th lymphocyte (HTL) epitopes cross-reactive with the murine I-A(b) class II molecule were used to evaluate different vaccine design strategies to simultaneously induce multiple HTL responses. All four epitopes were immunogenic in H-2(b) mice, demonstrating the feasibility of murine models to evaluate epitope-based vaccines destined for human use. Immunization with a pool of peptides induced responses against all four epitopes; illustrating immunodominance does not prevent the induction of balanced multispecific responses. When different delivery systems were evaluated, a multiple Ag peptide construct was found to be less efficient than a linear polypeptide encompassing all four epitopes. Further characterization of linear polypeptide revealed that the sequential arrangement of the epitopes created a junctional epitope with high affinity class II binding. Disruption of this junctional epitope through the introduction of a GPGPG spacer restored the immunogenicity against all four epitopes. Finally, we demonstrate that a GPGPG spacer construct can be used to induce HTL responses by either polypeptide or DNA immunization, highlighting the flexibility of the approach. C1 Epimmune, San Diego, CA 92121 USA. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Livingston, B (reprint author), Epimmune, 5820 Nancy Ridge Dr,Suite 100, San Diego, CA 92121 USA. FU NIAID NIH HHS [1-R44 AI49051-01] NR 35 TC 85 Z9 94 U1 0 U2 7 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD JUN 1 PY 2002 VL 168 IS 11 BP 5499 EP 5506 PG 8 WC Immunology SC Immunology GA 555WA UT WOS:000175815200016 PM 12023344 ER PT J AU Mariner, JM Mamane, Y Hiscott, J Waldmann, TA Azimi, N AF Mariner, JM Mamane, Y Hiscott, J Waldmann, TA Azimi, N TI IFN regulatory factor 4 participates in the human T cell lymphotropic virus type I-mediated activation of the IL-15 receptor alpha promoter SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NF-KAPPA-B; INTERLEUKIN-2 RECEPTOR; LEUKEMIA-VIRUS; GENE-EXPRESSION; NATURAL-KILLER; BETA-CHAIN; TRANSCRIPTIONAL ACTIVATOR; BINDING-PROTEIN; HTLV-I; MICE AB IL-15Ralpha mRNA and protein levels are increased in human T cell lymphotropic virus type-I (HTLV-I)-associated adult T cell leukemia. Previously, we demonstrated that IL-15Ralpha expression was activated by HTLV-1 Tax, in part, through the action of NF-kappaB. However, there appeared to be additional motifs within the IL-15Ralpha promoter that were responsive to HTLV-I Tax. In this study, we demonstrated that IL-15Ralpha mRNA expression was activated in human monocytes by IFN treatment, suggesting a role for IFN regulatory factors (IRFs) in IL-15Ralpha transcription. In addition, an IRF element within the Tax-responsive element of the IL-15Ralpha promoter was necessary for maximal Tax-induced activation of this promoter. Furthermore, we demonstrated that IRF-4, a transcription factor known to be elevated in HTLV-I-infected cells, activated the IL-15Ralpha promoter. Inhibition of IRF-4 action lead to reduced Tax-induced activation of the IL-15Ralpha promoter, while inhibition of both IRF-4 and NF-kappaB severely inhibited the Tax-induced activation of this promoter. These findings suggest a role for both NF-kappaB and IRF-4 in the transcriptional regulation of IL-15Ralpha by HTLV-I Tax. It is possible that the HTLV-I Tax-mediated induction of IL-15Ralpha and IL-15 may lead to an autocrine cytokine-mediated stimulatory loop leading to the proliferation of HTLV-1 infected cells. This loop of proliferation may facilitate viral propagation and play a role in HTLV-1-mediated disease progression. C1 NCI, Ctr Canc Res, Metab Branch, NIH, Bethesda, MD 20892 USA. Lady Davis Inst Med Res, Terry Fox Mol Oncol Grp, Montreal, PQ, Canada. McGill Univ, Dept Microbiol & Immunol, Montreal, PQ, Canada. McGill Univ, Dept Med, Montreal, PQ, Canada. McGill Univ, Dept Oncol, Montreal, PQ, Canada. RP Azimi, N (reprint author), NCI, Ctr Canc Res, Metab Branch, NIH, 10 Ctr Dr,MSC 1374,Bldg 10,Room 4N-102, Bethesda, MD 20892 USA. NR 43 TC 20 Z9 20 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 JUN 1 PY 2002 VL 168 IS 11 BP 5667 EP 5674 PG 8 WC Immunology SC Immunology GA 555WA UT WOS:000175815200037 PM 12023365 ER PT J AU Choi, WS Kim, YM Combs, C Frohman, MA Beaven, MA AF Choi, WS Kim, YM Combs, C Frohman, MA Beaven, MA TI Phospholipases D1 and D2 regulate different phases of exocytosis in mast cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-KINASE-C; ADP-RIBOSYLATION FACTOR; MUSCARINIC ACETYLCHOLINE-RECEPTOR; PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE; PHOSPHATIDIC-ACID; DEPENDENT ACTIVATION; RBL-2H3 CELLS; GENE FAMILY; RAT-BRAIN; D DEFINES AB The rat mast cell line RBL-2H3 contains both phospholipase D (PLD)1 and PLD2. Previous studies with this cell line indicated that expressed PLD1 and PLD2 are both strongly activated by stimulants of secretion. We now show by use of PLDs tagged with enhanced green fluorescent protein that PLD1, which is largely associated with secretory granules, redistributes to the plasma membrane in stimulated cells by processes reminiscent of exocytosis and fusion of granules with the plasma membrane. These processes and secretion of granules are suppressed by expression of a catalytically inactive mutant of PLD1 or by the presence of 50 mM 1-butanol but not tent-butanol, an indication that these events are dependent on the catalytic activity of PLD1. Of note, cholera toxin induces translocation of PLD1-labeled granules to the plasma membrane but not fusion of granules with plasma membrane or secretion. Subsequent stimulation of calcium influx with Ag or thapsigargin leads to rapid redistribution of PLD1 to the plasma membrane and accelerated secretion. Also of note, PLD1 is recycled from plasma membrane back to granules within 4 h of stimulation. PLD2, in contrast, is largely confined to the plasma membrane, but it too participates in the secretory process, because expression of catalytically inactive PLD2 also blocks secretion. These data indicate a two-step process: translocation of granules to the cell periphery, regulated by granule-associated PLD1, and a calcium-dependent fusion of granules with the plasma membrane, regulated by plasma membrane-associated PLD2 and possibly PLD1. C1 NHLBI, Lab Mol Immunol, NIH, Bethesda, MD 20892 USA. NHLBI, Light Microscopy Core Facil, NIH, Bethesda, MD 20892 USA. SUNY Stony Brook, Dept Pharmacol, Stony Brook, NY 11794 USA. SUNY Stony Brook, Ctr Dev Genet, Stony Brook, NY 11794 USA. RP Beaven, MA (reprint author), NHLBI, Lab Mol Immunol, NIH, Room 8N109,Bldg 10, Bethesda, MD 20892 USA. NR 46 TC 94 Z9 95 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 JUN 1 PY 2002 VL 168 IS 11 BP 5682 EP 5689 PG 8 WC Immunology SC Immunology GA 555WA UT WOS:000175815200039 PM 12023367 ER PT J AU Monsurro, V Nagorsen, D Wang, E Provenzano, M Dudley, ME Rosenberg, SA Marincola, FM AF Monsurro, V Nagorsen, D Wang, E Provenzano, M Dudley, ME Rosenberg, SA Marincola, FM TI Functional heterogeneity of vaccine-induced CD8(+) T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID IN-VITRO STIMULATION; MELANOMA PATIENTS; METASTATIC MELANOMA; PERIPHERAL-BLOOD; ANTIGENIC PEPTIDE; TUMOR-REGRESSION; DENDRITIC CELLS; REACTIVE CTL; GENE MAGE-3; EXPRESSION AB The functional status of circulating vaccine-induced, tumor-specific T cells has been questioned to explain their paradoxical inability to inhibit tumor growth. We enumerated with HLA-A*0201/peptide tetramers (tHLA) vaccine-elicited CD8(+) T cell precursor frequency among PBMC in 13 patients with melanoma undergoing vaccination with the HLA-A*0201-associated gp100: 209-217(210 M) epitope. T cell precursor frequency increased from undetectable to 12,400 +/- 3,600 x 105 CD8+ T cells after vaccination and appeared heterogeneous according to previously described functional subtypes: CD45RA(+)CD27(+) (14 +/- 2.6% of tHLA-staining T cells), naive; CD4RA(-)CD27(+) (14 +/- 3.2%), memory; CD45RA(+)CD27(-) (43 +/- 6%), effector; and CD45RA(-)CD27(-) (30 +/- 4.1%), memory/effector. The majority of tHLA(+)CD8(+) T cells displayed an effector, CD27(-) phenotype (73%). However, few expressed perforin (17%). Epitope-specific in vitro stimulation (IVS) followed by 10-day expansion in IL-2 reversed this phenotype by increasing the number of perforin(+) (84 +/- 3.6%; by paired t test, p < 0.001) and CD27(+) (from 28 to 67%; by paired t test, p = 0.01) tHLA(+) T cells. This conversion probably represented a change in the functional status of tHLA(+) T cells rather than a preferential expansion of a CD27(+) (naive and/or memory) PBMC, because it was reproduced after IVS of a T cell clone bearing a classic effector phenotype (CD45RA(+)CD27(-)). These findings suggest that circulating vaccine-elicited T cells are not as functionally active as inferred by characterization of IVS-induced CTL. In addition, CD45RA/CD27 expression may be more informative about the status of activation of circulating T cells than their status of differentiation. C1 NCI, Immunogenet Sect, Dept Transfus Med, Ctr Clin,NIH, Bethesda, MD 20892 USA. NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Marincola, FM (reprint author), NCI, Immunogenet Sect, Dept Transfus Med, Ctr Clin,NIH, Bldg 10,Room 1C-711,10 Ctr Dr,MSC 1502, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z01 SC003811-33] NR 54 TC 73 Z9 76 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 JUN 1 PY 2002 VL 168 IS 11 BP 5933 EP 5942 PG 10 WC Immunology SC Immunology GA 555WA UT WOS:000175815200072 PM 12023400 ER PT J AU Purcell, RH Wong, DC Shapiro, M AF Purcell, RH Wong, DC Shapiro, M TI Relative infectivity of hepatitis A virus by the oral and intravenous routes in 2 species of nonhuman primates SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID A VIRUS; VACCINE AB Hepatitis A virus (HAV) is naturally transmitted by the fecal-oral route but can also be transmitted intravenously. To determine the relative infectivity of these 2 routes, an infectivity titration of a standard challenge pool of virulent HAV was performed in tamarins and chimpanzees. In both species, 1 oral dose of HAV was equivalent to 10(4.5) intravenous doses. These findings have relevance for attempts to develop live, attenuated HAV vaccines that can be administered orally. C1 NIAID, Hepatitis Viruses Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. Bioqual, Rockville, MD USA. RP Purcell, RH (reprint author), NIAID, Hepatitis Viruses Sect, Infect Dis Lab, NIH, 50 South Dr,MSC 8009,Bldg 50,Rm 8523, Bethesda, MD 20892 USA. FU NIAID NIH HHS [N01 AI 52705] NR 15 TC 18 Z9 19 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUN 1 PY 2002 VL 185 IS 11 BP 1668 EP 1671 DI 10.1086/340520 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 556LK UT WOS:000175850800018 PM 12023776 ER PT J AU Chun, TW Justement, JS Pandya, P Hallahan, CW McLaughlin, M Liu, SY Ehler, LA Kovacs, C Fauci, AS AF Chun, TW Justement, JS Pandya, P Hallahan, CW McLaughlin, M Liu, SY Ehler, LA Kovacs, C Fauci, AS TI Relationship between the size of the human immunodeficiency virus type 1 (HIV-1) reservoir in peripheral blood CD4(+) T cells and CD4(+): CD8(+) T cell ratios in aviremic HIV-1-Infected individuals receiving long-term highly active antiretroviral therapy SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID LYMPHOCYTE-ACTIVATION; PLASMA VIREMIA; RNA LEVELS; INFECTION; REPLICATION; PERSISTENCE AB It has been demonstrated that human immunodeficiency virus type 1 (HIV-1) replication persists in most infected individuals receiving highly active antiretroviral therapy (HAART). However, studies addressing the relationship between low levels of ongoing viral replication and immunologic parameters, such as the CD4(+):CD8(+) T cell ratio, in such individuals have been lacking. Here, a statistically significant inverse correlation is shown between the frequency of CD4(+) T cells carrying HIV-1 proviral DNA and the CD4(+):CD8(+) T cell ratio in infected individuals receiving HAART and in whom plasma viremia had been suppressed below the limit of detection for prolonged periods of time. No correlation was found between the frequency of HIV-1-specific cytotoxic CD8(+) T lymphocytes (CTLs) and the CD4(+):CD8(+) T cell ratios in those individuals. These data suggest that persistent, low-level, ongoing viral replication, although not sufficient to maintain HIV-1 specific CTL responses, may explain, in part, why normalization of the CD4(+):CD8(+) T cell ratio is not achieved in some infected individuals successfully treated with HAART. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. Univ Toronto, Dept Med, Toronto, ON, Canada. RP Chun, TW (reprint author), NIAID, Immunoregulat Lab, NIH, Bldg 10,Rm 6A32, Bethesda, MD 20892 USA. NR 15 TC 57 Z9 57 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUN 1 PY 2002 VL 185 IS 11 BP 1672 EP 1676 DI 10.1086/340521 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 556LK UT WOS:000175850800019 PM 12023777 ER PT J AU Kim, SY Jeong, EJ Steinert, PM AF Kim, SY Jeong, EJ Steinert, PM TI IFN-gamma induces transglutaminase 2 expression in rat small intestinal cells SO JOURNAL OF INTERFERON AND CYTOKINE RESEARCH LA English DT Article ID CELIAC-DISEASE; TISSUE TRANSGLUTAMINASE; INTERFERON-GAMMA; CROSS-LINKING; T-CELLS; APOPTOSIS; GLIADIN; BETA; IDENTIFICATION; AUTOANTIGEN AB Transglutaminase 2 (tissue transglutaminase, TGase 2) was recently identified as an endomysial autoantigen in celiac disease (CD). Identification of how TGase 2 expression is increased may allow a better understanding of this autoimmune disease. Certain inflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta), and the Th type I cytokine interferon-gamma (INF-gamma) are abundant in CD. We have investigated whether these play a role in the regulation of TGase 2 expression in a model rat small intestinal epithelial cell line (IEC-6). After treatment for 24 h, TNF-alpha did not significantly alter TGase 2 mRNA or activity, but TGF-beta decreased mRNA and activity by 4-5-fold. IFN-gamma increased mRNA and TGase 2 activity by about 2-fold in 24 h and 5-fold by 5 days. Our new data suggest that increased TGase 2 expression in the upper small intestine of CD patients may be due to increased IFN-gamma expression, loss of TGF-beta signaling, or both. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. RP Kim, SY (reprint author), Cornell Univ, Weill Med Coll, Burke Med Res Inst, 785 Mamaroneck Ave, White Plains, NY 10605 USA. NR 31 TC 32 Z9 32 U1 0 U2 0 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 JUN PY 2002 VL 22 IS 6 BP 677 EP 682 DI 10.1089/10799900260100169 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Immunology SC Biochemistry & Molecular Biology; Cell Biology; Immunology GA 568QM UT WOS:000176553900008 PM 12162878 ER PT J AU Emmert, S Slor, H Busch, DB Batko, S Albert, RB Coleman, D Khan, SG Abu-Libdeh, B DiGiovanna, JJ Cunningham, BB Lee, MM Crollick, J Inui, H Ueda, T Hedayati, M Grossman, L Shahlavi, T Cleaver, JE Kraemer, KH AF Emmert, S Slor, H Busch, DB Batko, S Albert, RB Coleman, D Khan, SG Abu-Libdeh, B DiGiovanna, JJ Cunningham, BB Lee, MM Crollick, J Inui, H Ueda, T Hedayati, M Grossman, L Shahlavi, T Cleaver, JE Kraemer, KH TI Relationship of neurologic degeneration to genotype in three xeroderma pigmentosum group G patients SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE Cockayne syndrome; DNA repair; host cell reactivation; molecular genetics; sun sensitivity ID NUCLEOTIDE EXCISION-REPAIR; COMPLEMENTATION GROUP-G; TRANSCRIPTION-COUPLED REPAIR; COCKAYNE-SYNDROME PATIENTS; DNA-REPAIR; XPG PROTEIN; SKIN-CANCER; SACCHAROMYCES-CEREVISIAE; YEAST RAD2; GENE AB We studied three newly diagnosed xeroderma pigmentosum complementation group G patients with markedy different clinical features. An Israeli-Palestinian girl (XP96TA) had severe abnormal ities suggestive of the xeroderma pigmentosum/Cockayne syndrome complex including sun sensitivity, neurologic and developmental impairment, and death by age 6 y. A Caucasian girl (XP82DC) also had severe sun sensitivity with neurologic and developmental impairment and died at 5.8 y. In contrast, a mildly affected 14-y-old Caucasian female (XP65BE) had sun sensitivity but no neurologic abnormalities. XP96TA, XP82DC, and XP65BE fibroblasts showed marked reductions in post-ultraviolet cell survival and DNA repair but these were higher in XP65BE than in XP82DC. XP96TA fibroblasts had very low XPG mRNA expression levels whereas XP65BE fibroblasts had nearly normal levels. Host cell reactivation of an ultraviolet-treated reporter assigned all three fibroblast strains to the rare xeroderma pigmentosum complementation group G (only 10 other patients previously reported). XP96TA and XP82DC cells had mutations in both XPG alleles that are predicted to result in severely truncated proteins including stop codons and two base frameshifts. The mild XP65BE patient had an early stop codon mutation in the paternal allele. The XP65BE maternal allele had a single base missense mutation (G2817A, Ala874Thr) that showed residual ability to complement xeroderma pigmentosum complementation group G cells. These observations agree with earlier studies demonstrating that XPG mutations, which are predicted to lead to severely truncated proteins in both alleles, were associated with severe xeroderma pigmentosum/Cockayne syndrome neurologic symptoms. Retaining residual functional activity in one allele was associated with mild clinical features without neurologic abnormalities. C1 NCI, Basic Res Lab, Canc Res Ctr, Bethesda, MD 20892 USA. Tel Aviv Univ, Sackler Sch Med, Dept Human Genet & Mol Med, IL-69978 Tel Aviv, Israel. Armed Forces Inst Pathol, Dept Environm & Toxicol Pathaol, Washington, DC 20306 USA. Makassed Islam Charitable Hosp, Jerusalem, Israel. Brown Univ, Sch Med, Dept Dermatol, Providence, RI 02912 USA. Univ Calif San Diego, San Diego, CA 92103 USA. Childrens Hosp & Hlth Ctr, Dept Dermatol, San Diego, CA USA. Kaiser Permanente, Dermatol Sect, Downey, CA USA. SUNY Buffalo, Dept Dermatol, Buffalo, NY 14260 USA. Johns Hopkins Univ, Sch Publ Hlth, Dept Biochem, Baltimore, MD USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. RP Kraemer, KH (reprint author), NCI, Basic Res Lab, Canc Res Ctr, Bldg 37,Room 3E24, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z01 BC004517-31] NR 66 TC 63 Z9 65 U1 0 U2 1 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD JUN PY 2002 VL 118 IS 6 BP 972 EP 982 DI 10.1046/j.1523-1747.2002.01782.x PG 11 WC Dermatology SC Dermatology GA 559NA UT WOS:000176031200010 PM 12060391 ER PT J AU Domachowske, JB Bonville, CA Easton, AJ Rosenberg, HF AF Domachowske, JB Bonville, CA Easton, AJ Rosenberg, HF TI Pulmonary eosinophilia in mice devoid of interleukin-5 SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article ID RESPIRATORY SYNCYTIAL VIRUS; AIRWAY HYPERRESPONSIVENESS; HOST-DEFENSE; RIBONUCLEASE GENES; INFECTED MICE; LUNG; IL-5; INFLAMMATION; EXPRESSION; ASTHMA AB The biology of the eosinophilic leukocyte-development, recruitment, and prolonged existence in somatic tissues has been linked almost invariably to the actions of the "eosinophil" cytokine, interleukin-5 (IL-5). Here we demonstrate that pulmonary eosinophilia can occur in the absence of IL-5, as morphologically normal eosinophils are recruited to the lungs of virus-infected IL-5 -/- mice with kinetics and sequelae that are indistinguishable from those of their IL-5 +/+ Counterparts. We conclude that pulmonary eosinophilia observed in response to primary paramyxovirus infection occurs via mechanisms that are distinct from those involved in eosinophil responses to allergens and in asthma. Furthermore, the presence of functional eosinophils in IL-5 -/- mice suggests the possibility of developmentally distinct subsets of what has been presumed to be a homogeneous leukocyte population. C1 NIAID, LHD, Eosinophil Pathophysiol Sect, NIH, Bethesda, MD 20892 USA. SUNY Syracuse, Upstate Med Univ, Dept Pediat, Syracuse, NY USA. Univ Warwick, Dept Biol, Coventry CV4 7AL, W Midlands, England. RP Rosenberg, HF (reprint author), NIAID, LHD, Eosinophil Pathophysiol Sect, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 38 TC 13 Z9 15 U1 0 U2 2 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD JUN PY 2002 VL 71 IS 6 BP 966 EP 972 PG 7 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 560JG UT WOS:000176078500007 PM 12050181 ER PT J AU Ardans, JA Economou, AP Martinson, JM Zhou, M Wahl, LM AF Ardans, JA Economou, AP Martinson, JM Zhou, M Wahl, LM TI Oxidized low-density and high-density lipoproteins regulate the production of matrix metalloproteinase-1 and-9 by activated monocytes SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE atherosclerosis; prostaglandins; TNF-alpha; GM-CSF ID HUMAN ATHEROSCLEROTIC PLAQUES; MACROPHAGE; EXPRESSION; CYTOKINES; PROSTACYCLIN; COLLAGENASE; MECHANISMS; RECEPTOR; REGIONS; LESIONS AB Monocytes/macrophages are prominent in atherosclerotic plaques where the vascular remodeling and plaque rupture may be influenced by the lipids and cytokines at these sites. Therefore, we evaluated the effects of factors found,within the vascular wall, such as cytokines, Oxidized low-density lipoprotein (ox-LDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL), on monocyte-derived matrix metalloproteinase-1 (MMP-1) and -9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1). ox-LDL, LDL, and HDL alone had no effect on MMP-1, MMP-9, or TIMP-1 production. However, in the presence of tumor necrosis factor (TNF)-alpha and GM-CSF, ox-LDL enhanced MMP-1 significantly by two- to threefold, increased MMP-9 slightly, and had no effect on TIMP-1 production. In contrast, HDL suppressed the induction of MMP-1 by TNF-alpha and GM-CSF as well as the ox-LDL-mediated increase in MMP-1 production. The enhancement of MMP-1 production by ox-LDL occurred through, in part, a prostaglandin E-2 (PGE(2))-dependent pathway as indomethacin suppressed and PGE(2) restored MMP-1 production. This conclusion was supported further by ox-LDL-mediated increases in PGE(2) and cyclooxygenase-2 (COX-2) production. These data suggest that the interaction of primary monocytes with ox-LDL and proinflammatory cytokines may contribute to vascular remodeling and plaque rupture. C1 NIDCR, Immunopathol Sect, NIH, Bethesda, MD 20892 USA. RP Wahl, LM (reprint author), NIDCR, Immunopathol Sect, NIH, Bldg 30,Room 325, Bethesda, MD 20892 USA. NR 32 TC 48 Z9 54 U1 1 U2 1 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD JUN PY 2002 VL 71 IS 6 BP 1012 EP 1018 PG 7 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 560JG UT WOS:000176078500013 PM 12050187 ER PT J AU Galban, CJ Spencer, RGS AF Galban, CJ Spencer, RGS TI Optimized pulse parameters for reducing quantitation errors due to saturation factor changes in magnetic resonance spectroscopy SO JOURNAL OF MAGNETIC RESONANCE LA English DT Article ID LATTICE RELAXATION-TIMES; CHEMICAL-EXCHANGE; IN-VIVO; MUSCLE; NMR; SYSTEMS; HEART AB We present an analysis of the effects of chemical exchange and changes in T-1 on metabolite quantitation for heart, skeletal muscle, and brain using the one-pulse experiment for a sample which is subject to temporal variation. We use an optimization algorithm to calculate interpulse delay times, TRs, and flip angles, theta, resulting in maximal root-mean-squared signal-to-noise per unit time (SIN) for all exchanging species under 5 and 10% constraints on quantitation errors. The optimization yields TR and theta pairs giving signal-to-noise per unit time close or superior to typical literature values. Additional simulations were performed to demonstrate explicitly the dependence of the quantitation errors on pulse parameters and variations in the properties of the sample, such as may occur after an intervention. We find that (i) correction for partial saturation in accordance with the usual analysis neglecting variations in metabolite concentrations and rate constants may readily result in quantitation errors of 15% or more; the exact degree of error depends upon the details of the system under consideration; (ii) if T-1's vary as well, significantly larger quantitation errors may occur; and (iii) optimal values of pulse parameters may minimize errors in quantitation with minimal S/N loss. (C) 2002 Elsevier Science (USA). C1 NIA, NIH, Baltimore, MD 21224 USA. RP Spencer, RGS (reprint author), NIA, NIH, GRC 4D-08,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 15 TC 5 Z9 5 U1 1 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1090-7807 J9 J MAGN RESON JI J. Magn. Reson. PD JUN PY 2002 VL 156 IS 2 BP 161 EP 170 DI 10.1006/jmre.2002.2542 PG 10 WC Biochemical Research Methods; Physics, Atomic, Molecular & Chemical; Spectroscopy SC Biochemistry & Molecular Biology; Physics; Spectroscopy GA 584RB UT WOS:000177480900001 PM 12165250 ER PT J AU Ding, KY Gronenborn, AM AF Ding, KY Gronenborn, AM TI Novel 2D triple-resonance NMR experiments for sequential resonance assignments of proteins SO JOURNAL OF MAGNETIC RESONANCE LA English DT Article DE proteins; sequential assignment; reduced dimensionality; structural genomics ID BIOLOGICAL MACROMOLECULES; REDUCED-DIMENSIONALITY; BACKBONE ASSIGNMENT; LARGER PROTEINS; PEAK PICKING; SPECTROSCOPY; SPECTRA; RESOLUTION; RELAXATION AB We present 2D versions of the popular triple resonance HN(CO) CACB, HN(COCA)CACB, HN(CO)CAHA, and HN(COCA) CARA experiments, commonly used for sequential resonance assignments of proteins. These experiments provide information about correlations between amino proton and nitrogen chemical shifts and the alpha- and beta-carbon and alpha-proton chemical shifts within and between amino acid residues. Using these 2D spectra, sequential resonance assignments of H-N, N, C-alpha, C-beta, and H-alpha nuclei are easily achieved. The resolution of these spectra is identical to the well-resolved 2D N-15-H-1 HSQC and H(NCO)CA spectra, with slightly reduced sensitivity compared to their 3D and 4D versions. These types of spectra are ideally suited for exploitation in automated assignment procedures and thereby constitute a fast and efficient means for NMR structural determination of small and medium-sized proteins in solution in structural genomics programs. (C) 2002 Elsevier Science (USA). C1 NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Gronenborn, AM (reprint author), NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. OI Gronenborn, Angela M/0000-0001-9072-3525 NR 37 TC 93 Z9 96 U1 0 U2 6 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1090-7807 J9 J MAGN RESON JI J. Magn. Reson. PD JUN PY 2002 VL 156 IS 2 BP 262 EP 268 DI 10.1006/jmre.2002.2537 PG 7 WC Biochemical Research Methods; Physics, Atomic, Molecular & Chemical; Spectroscopy SC Biochemistry & Molecular Biology; Physics; Spectroscopy GA 584RB UT WOS:000177480900013 PM 12165262 ER PT J AU St Lawrence, KS Ye, FQ Lewis, BK Weinberger, DR Frank, JA McLaughlin, AC AF St Lawrence, KS Ye, FQ Lewis, BK Weinberger, DR Frank, JA McLaughlin, AC TI Effects of indomethacin on cerebral blood flow at rest and during hypercapnia: An arterial spin tagging study in humans SO JOURNAL OF MAGNETIC RESONANCE IMAGING LA English DT Article; Proceedings Paper CT 9th Scientific Meeting of the ISMRM CY APR 21-27, 2001 CL GLASGOW, SCOTLAND SP ISMRM DE cerebral blood flow; hypercapnia; arterial spin tagging; arterial spin labeling; indomethacin ID CARBON-DIOXIDE REACTIVITY; FUNCTIONAL MRI; PROSTAGLANDIN SYNTHESIS; REGIONAL DIFFERENCES; OXYGEN-CONSUMPTION; NITROUS-OXIDE; STEP CHANGES; ACTIVATION; PERFUSION; BRAIN AB Purpose: To investigate using an arterial spin tagging (AST) approach the effect of indomethacin on the cerebral blood flow (CBF) response to hypercapnia. Materials and Methods: Subjects inhaled a gas mixture containing 6% CO2 for two 5-minute periods, which were separated by a 10-minute interval, in which subjects inhaled room air. In six subjects, indomethacin (i.v., 0.2 mg/ kg) was infused in the normocapnic interval between the two hypercapnic periods. Results: Indomethacin reduced normocapnic gray matter CBF by 36 +/- 5% and reduced the CBF increase during hypercapnia from 43 +/- 9% to 16 +/- 5% in gray matter (P < 0.001) and from 48 +/- 11% to 35 +/- 9% in white matter (P < 0.025). Conclusion: The results demonstrate that an AST approach can measure the effects of indomethacin on global CBF increases during hypercapnia and suggest that an AST approach could be used to investigate pharmacological effects on focal CBF increases during functional activation. C1 NIH, Lab Diagnost Radiol Res, Clin Ctr, Bethesda, MD 20892 USA. RP St Lawrence, KS (reprint author), NIH, Lab Diagnost Radiol Res, Clin Ctr, Bldg 10 Rm B1N 256 10 Ctr Dr MSC 1074, Bethesda, MD 20892 USA. RI St. Lawrence, Keith/B-5726-2015 NR 40 TC 17 Z9 17 U1 0 U2 1 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 1053-1807 J9 J MAGN RESON IMAGING JI J. Magn. Reson. Imaging PD JUN PY 2002 VL 15 IS 6 BP 628 EP 635 DI 10.1002/jmri.10111 PG 8 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 557QB UT WOS:000175918300002 PM 12112512 ER PT J AU Clay, JR Shrier, A AF Clay, JR Shrier, A TI Temperature dependence of bistability in squid giant axons with alkaline intracellular pH SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE nerve; automaticity; alkalinity; action potential; temperature; ion; channel gating ID CONDUCTANCE; MEMBRANE; CHANNEL; NEURONS; MODEL; ANNIHILATION; CURRENTS; APNEA AB Raising the intracellular pH (pH(i)) above 7.7 in intracellularly perfused squid giant axons causes spontaneous firing of action potentials. The firing frequency ranged from 20 Hz at 0degreesC to 200 Hz at 23degreesC. Above 23degreesC, the axons were quiescent. They were bistable for 13 < T < 23degreesC. That is, they were either quiescent or spontaneously firing. Below 13degreesC, spontaneous firing was the only stable element. The primary effects of changes in temperature on the underlying ionic currents were on gating of the delayed rectifier potassium channel I-K, and the sodium ion channel I-Na. The kinetics of I-K had a Q(10) of 3.63. The effect of T on I-Na was more complicated in that the peak I-Na amplitude increased with T, as demonstrated in earlier reports. This effect, as well as the changes in I-Na kinetics produced by changes in T, were mimicked in the context of a model of I-Na gating in which activation and inactivations are coupled. Electrical activity was simulated in a model of the action potential with appropriate temperature-dependent modifications for I-Na and I-K. The model predicts a change from monostability (spontaneous firing) at relatively low temperatures to bistability (quiescence and spontaneous firing) as the temperature is raised, followed by change back to monostability (quiescence) as the temperature is further increased, which is consistent with the experimental results. C1 NINCDS, Ion Channel Biophys Unit, Div Intramural Res, NIH, Bethesda, MD 20892 USA. McGill Univ, Dept Physiol, Montreal, PQ H3G 1Y6, Canada. RP Clay, JR (reprint author), NINCDS, Ion Channel Biophys Unit, Div Intramural Res, NIH, Bethesda, MD 20892 USA. NR 30 TC 8 Z9 8 U1 0 U2 0 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 JUN 1 PY 2002 VL 187 IS 3 BP 213 EP 223 DI 10.1007/s00232-001-0165-3 PG 11 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 561AQ UT WOS:000176114100005 PM 12163979 ER PT J AU Janczewski, AM Spurgeon, HA Lakatta, EG AF Janczewski, AM Spurgeon, HA Lakatta, EG TI Action potential prolongation in cardiac myocytes of old rats is an adaptation to sustain youthful intracellular Ca2+ regulation SO JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY LA English DT Article DE action potential-clamp; L-type Ca2+ current; excitation-contraction coupling; Ca-i(2+) transient; sarcoplasmic reticulum; caffeine; Na+-Ca2+ exchange; Ca2+-induced Ca2+ release gain ID SARCOPLASMIC-RETICULUM CA2+-ATPASE; PIG VENTRICULAR MYOCYTES; AGE-ASSOCIATED CHANGES; EXCITATION-CONTRACTION; GUINEA-PIG; NA+-CA2+ EXCHANGE; CALCIUM; HEART; RELEASE; CELLS AB Advanced age in rats is accompanied by reduced expression of the sarcoplasmic reticulum (SR) Ca2+ pump (SERCA-2). The amplitudes of intracellular Ca2+ (Ca+) transients and contractions in ventricular myocytes isolated from old (23-24-months) rats (OR), however, are similar to those of young (4-6-months) rat myocytes (YR). OR myocytes also manifest slowed inactivation of L-type Ca2+ current (I-CaL) and marked prolongation of action potential (AP) duration. To determine whether and how age-associated AP prolongation preserves the Ca-i(2+) transient amplitude in OR myocytes, we employed an AP-clamp technique with simultaneous measurements of I-CaL (with Na+ current, K+ currents and Ca2+ influx via sarcolemmal Na+-Ca2+ exchanger blocked) and Ca-i(2+) transients in OR rat ventricular myocytes dialyzed with the fluorescent Ca2+ probe, indo-1. Myocytes were stimulated with AP-shaped voltage clamp waveforms approximating the configuration of prolonged, i.e. the native, AP of OR cells (AP-L), or with short AP waveforms (AP-S), typical of YR myocytes. Changes in SR Ca2+ load were assessed by rapid, complete SR Ca2+ depletions with caffeine. As expected, during stimulation with AP-S vs AP-L, peak I-CaL increased, by 21 +/- 4%, while the I-CaL integral decreased, by 19 +/- 3% (P < 0.01 for each). Compared to AP-L, stimulation of OR myocytes with AP-S reduced the amplitudes of the Ca-i(2+) transient by 31 +/- 6%, its maximal rate of rise (+ dCa(i)(2+)/dt(max); a sensitive index of SR Ca2+ release flux) by 37 +/- 4%, and decreased the SR Ca2+ load by 29 4% (P < 0.01 for each). Intriguingly, AP-S also reduced the maximal rate of the Ca2+ transient relaxation and prolonged its time to 50% decline, by 35 +/- 5% and 33 +/- 7%, respectively (P < 0.01 for each). During stimulation with AP-S, the gain of Ca2+-induced Ca2+ release (CICR), indexed by + dCa(i)(2+)/dt(max)/I-CaL, was reduced by 46 4% vs AP-L (P < 0.01). We conclude that the effects of an application of a shorter AP to OR myocytes to reduce +dCa(i)(2+)/dt(max) and the Ca2+ transient amplitude are attributable to a reduction in SR Ca2+ load, presumably due to a reduced I-CaL integral and likely also to an increased Ca2+ extrusion via sarcolemmal Na+-Ca2+ exchanger. The decrease in the Ca-i(2+) transient relaxation rate in OR cells stimulated with shorter APs may reflect a reduction of Ca2+/calmodulin-kinase II-regulated modulation of Ca2+ uptake via SERCA-2, consequent to a reduced local Ca2+ release in the vicinity of SERCA-2, also attributable to reduced SR Ca2+ load. Thus, the reduction of CICR gain during stimulation with AP-S is the net result of both a diminished SR Ca2+ release and an increased peak I-CaL. These results suggest that ventricular myocytes of old rats utilize AP prolongation to preserve an optimal SR Ca2+ loading, CICR gain and relaxation of Ca-i(2+) transients. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, Intramural Res Program,NIH, Baltimore, MD 21224 USA. RP Lakatta, EG (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, Intramural Res Program,NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. EM lakattae@grc.nia.nih.gov NR 41 TC 40 Z9 40 U1 0 U2 1 PU ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2828 EI 1095-8584 J9 J MOL CELL CARDIOL JI J. Mol. Cell. Cardiol. PD JUN PY 2002 VL 34 IS 6 BP 641 EP 648 DI 10.1006/jmcc.2002.2004 PG 8 WC Cardiac & Cardiovascular Systems; Cell Biology SC Cardiovascular System & Cardiology; Cell Biology GA 567QW UT WOS:000176496800007 PM 12054851 ER PT J AU Sollott, SJ AF Sollott, SJ TI Endogenous no mechanisms mediate the stretch dependence of Ca2+ release in cardiac myocytes. SO JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY LA English DT Meeting Abstract C1 NIA, IRP, NIH, Ctr Gerontol Res, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2828 EI 1095-8584 J9 J MOL CELL CARDIOL JI J. Mol. Cell. Cardiol. PD JUN PY 2002 VL 34 IS 6 BP A59 EP A59 DI 10.1016/S0022-2828(02)90961-2 PG 1 WC Cardiac & Cardiovascular Systems; Cell Biology SC Cardiovascular System & Cardiology; Cell Biology GA 567QW UT WOS:000176496800239 ER PT J AU Kittur, S Wilasrusmee, S Pedersen, WA Mattson, MP Straube-West, K Wilasrusmee, C Jubelt, B Kittur, DS AF Kittur, S Wilasrusmee, S Pedersen, WA Mattson, MP Straube-West, K Wilasrusmee, C Jubelt, B Kittur, DS TI Neurotrophic and neuroprotective effects of milk thistle (Silybum marianum) on neurons in culture SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE apoptosis; herbal medicine; hippocampus; neurite outgrowth; oxidative stress ID CELL-DEATH; SILYMARIN; INHIBITION; APOPTOSIS; ANTIOXIDANT; ACTIVATION; EXPRESSION; SILIBININ; PATHWAY; PROTEIN AB Herbal products are being increasingly used as dietary supplements and therapeutic agents. However, much more research must be performed in order to determine the biological basis for their putative clinical effects. We tested the effects of milk thistle (Silybum marianum) extract on the differentiation and survival of cultured neural cells. Milk thistle enhanced nerve growth factor (NGF)-induced neurite outgrowth in PC-12 neural cells and prolonged their survival in culture. Milk thistle extract also protected cultured rat hippocampal neurons against oxidative stress-induced cell death. Our data demonstrate that milk thistle extract can promote neuronal differentiation and survival, suggesting potential benefits of chemicals in this plant on the nervous system. C1 SUNY Upstate Med Univ, Dept Neurol, Syracuse, NY 13210 USA. SUNY Upstate Med Univ, Dept Surg, Syracuse, NY 13210 USA. VA Med Ctr, Dept Neurol, Syracuse, NY 13210 USA. NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Kittur, S (reprint author), SUNY Upstate Med Univ, Dept Neurol, Syracuse, NY 13210 USA. RI Mattson, Mark/F-6038-2012 NR 26 TC 13 Z9 13 U1 1 U2 3 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 JUN PY 2002 VL 18 IS 3 BP 265 EP 269 DI 10.1385/JMN:18:3:265 PG 5 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 557NX UT WOS:000175915600012 PM 12059045 ER PT J AU Milanowski, DJ Winter, REK Elvin-Lewis, MPF Lewis, WH AF Milanowski, DJ Winter, REK Elvin-Lewis, MPF Lewis, WH TI Geographic disitribution of three alkaloid chemotypes of Croton lechleri SO JOURNAL OF NATURAL PRODUCTS LA English DT Article AB Three known alkaloids, isoboldine (2), norisoboldine (1), and magnoflorine (8), have been isolated for the first time from Croton lechleri, a source of the wound healing latex "sangre de grado". An HPLC system was developed, and a large number of latex and leaf samples of C. lechleri from 22 sites in northern Peru and Ecuador were analyzed to gain an understanding of the natural variation in alkaloid content for the species. Up to six alkaloids were found to occur in the leaves including, in addition to those listed above, thaliporphine (3), glaucine (4), and taspine (9), whereas the latex contained only 9. Taspine (9) is the component that has been previously found to be responsible for the wound healing activity of C. lechleri latex, and its mean concentration throughout the range examined was found to be 9% of the latex by dry weight. In addition, three chemotypes are defined based on the alkaloid content of the leaves, and the geographic distribution of these chemotypes is discussed along with a quantitative analysis of the alkaloid content as a function of chemotype. C1 Washington Univ, Dept Biol, St Louis, MO 63130 USA. Univ Missouri, Dept Chem, St Louis, MO 63110 USA. RP Milanowski, DJ (reprint author), NCI, Mol Targets Drug Discivery Program, Ctr Canc Res, Bldg 562,Room 301, Frederick, MD 21702 USA. FU FIC NIH HHS [U01TW00331] NR 13 TC 31 Z9 34 U1 1 U2 10 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 JUN PY 2002 VL 65 IS 6 BP 814 EP 819 DI 10.1021/np000270-v PG 6 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 570YP UT WOS:000176688300003 PM 12088421 ER PT J AU Mi, QW Lantvit, D Reyes-Lim, E Chai, HY Zhao, WM Lee, IS Peraza-Sanchez, S Ngassapa, O Kardono, LBS Riswan, S Hollingshead, MG Mayo, JG Farnsworth, NR Cordell, GA Kinghorn, AD Pezzuto, JM AF Mi, QW Lantvit, D Reyes-Lim, E Chai, HY Zhao, WM Lee, IS Peraza-Sanchez, S Ngassapa, O Kardono, LBS Riswan, S Hollingshead, MG Mayo, JG Farnsworth, NR Cordell, GA Kinghorn, AD Pezzuto, JM TI Evaluation of the potential cancer chemotherapeutic efficacy of natural product isolates employing in vivo hollow fiber tests SO JOURNAL OF NATURAL PRODUCTS LA English DT Article ID NUDE-MICE; INHIBITORY ACTIVITY; AQUEOUS-SOLUTION; TUMOR-MODEL; IN-VITRO; POLYVINYLPYRROLIDONE; DERIVATIVES; GLYCOSIDES; CELLS; TAXOL AB The hollow fiber test has been developed for the preliminary in vivo assessment of cancer chemotherapeutic efficacy of selected natural products. Using this model, we have established growth conditions for HL-60, HUVEC, Ishikawa, KB, KB-V1, LNCaP, Lu1, MCF-7, Mel2, P-388, and SW626 cells implanted at the intraperitoneal (i.p.) and subcutaneous (s.c.) compartments of athymic mice. Five cytotoxic natural product isolates (2-6) were tested in this model, along with paclitaxel (taxol) (1). Among the compounds tested, dioscin (2) and 13-methoxy-15-oxozoapatlin (3) were found to be active, indicating their potential to function as cancer chemotherapeutic agents. On the other hand, ochraceolide A (4), alpha-lapachone (5), and 2-(1-hydroxyethyl)naphtha[2,3-b]furan-4,9-quinone (6), all of which were significantly cytotoxic to cultured mammalian cells, did not mediate significant responses with the hollow fiber model. In further xenograft studies using KB cells implanted at the subcutaneous site, compound 3 mediated a statistically significant response which was consistent with the response observed at the subcutaneous compartment in the hollow fiber tests. In sum, these studies illustrate the usefulness of the hollow fiber model in natural product drug discovery programs. Preliminary indications of potential therapeutic efficacy can be provided quickly at relatively low expense. Agents capable of mediating a response at the subcutaneous site would appear to warrant greatest attention. C1 Univ Illinois, Coll Pharm, Program Collaborat Res Pharmaceut Sci, Chicago, IL 60612 USA. Univ Illinois, Coll Pharm, Dept Med Chem & Pharmacognosy, Chicago, IL 60612 USA. NCI, Frederick Canc Res & Dev Ctr, Biol Testing Branch, Dev Therapeut Program,Div Canc Treatment & Diag, Frederick, MD 21701 USA. RP Pezzuto, JM (reprint author), Univ Illinois, Coll Pharm, Program Collaborat Res Pharmaceut Sci, Chicago, IL 60612 USA. OI Kinghorn, A. Douglas/0000-0002-6647-8707 FU NCI NIH HHS [U19 CA52956] NR 48 TC 60 Z9 67 U1 1 U2 6 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 JUN PY 2002 VL 65 IS 6 BP 842 EP 850 DI 10.1021/np010322w PG 9 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 570YP UT WOS:000176688300007 PM 12088425 ER PT J AU Marquez, BL Watts, KS Yokochi, A Roberts, MA Verdier-Pinard, P Jimenez, JI Hamel, E Scheuer, PJ Gerwick, WH AF Marquez, BL Watts, KS Yokochi, A Roberts, MA Verdier-Pinard, P Jimenez, JI Hamel, E Scheuer, PJ Gerwick, WH TI Structure and absolute stereochemistry of hectochlorin, a potent stimulator of actin assembly SO JOURNAL OF NATURAL PRODUCTS LA English DT Article ID CYANOBACTERIUM LYNGBYA-MAJUSCULA; CURACIN-A; ANTIMITOTIC AGENT; NATURAL PRODUCT; JASPLAKINOLIDE; METABOLITE; POLYMERIZATION; KINETICS; SPONGE; CELLS AB Hectochlorin (1) was isolated from marine isolates of Lyngbya majuscula collected from Hector Bay, Jamaica, and Boca del Drago Beach, Bocas del Toro, Panama. The planar structure was deduced by one-and two-dimensional NMR spectroscopy. X-ray crystallography was used to determine the absolute stereochemistry of hectochlorin as 2S,3S, 14S,22S. Hectochlorin is equipotent to jasplakinolide (5) in its ability to promote actin polymerization, but unlike jasplakinolide, is unable to displace a fluorescent phalloidin analogue from polymerized actin. In addition, hectochlorin shows both a unique profile of cytotoxicity by the COMPARE algorithm and potent inhibitory activity toward the fungus Candida albicans. Structurally, hectochlorin resembles dolabellin and the recently reported lyngbyabellin class of compounds. C1 Oregon State Univ, Coll Pharm, Corvallis, OR 97331 USA. Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA. Oregon State Univ, Dept Chem, Corvallis, OR 97331 USA. NCI, Screening Technol Branch, Dev Therapeut Program, Div Canc Treatment & Diag,NIH, Frederick, MD 21702 USA. Univ Hawaii Manoa, Dept Chem, Honolulu, HI 96822 USA. RP Gerwick, WH (reprint author), Oregon State Univ, Coll Pharm, Corvallis, OR 97331 USA. OI Verdier-Pinard, Pascal/0000-0002-6149-6578 FU NCI NIH HHS [CA 52955]; NIEHS NIH HHS [P30-ES03850] NR 25 TC 76 Z9 80 U1 2 U2 10 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 JUN PY 2002 VL 65 IS 6 BP 866 EP 871 DI 10.1021/np0106283 PG 6 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 570YP UT WOS:000176688300011 PM 12088429 ER PT J AU Lutz, RJ Warren, K Balis, F Patronas, N Dedrick, RL AF Lutz, RJ Warren, K Balis, F Patronas, N Dedrick, RL TI Mixing during intravertebral arterial infusions in an in vitro model SO JOURNAL OF NEURO-ONCOLOGY LA English DT Article DE arterial infusion; mixing; brain tumors ID BLOOD-BRAIN-BARRIER; DRUG DELIVERY; INVITRO MODEL; CHEMOTHERAPY; EFFICACY; TUMORS AB Regional delivery of drugs can offer a pharmacokinetic advantage in the treatment of localized tumors. One method of regional delivery is by intra-arterial infusion into the basilar/vertebral artery network that provides local access to infratentorial tumors, which are frequent locations of childhood brain cancers. Proper delivery of drug by infused solutions requires adequate mixing of the infusate at the site of infusion within the artery lumen. Our mixing studies with an in vitro model of the vertebral artery network indicate that streaming of drug solution is likely to occur at low, steady infusion rates of 2 ml/min. Streaming leads to maldistribution of drug to distal perfused brain regions and may result in toxic levels in some regions while concurrently yielding subtherapeutic levels in adjacent regions. According to our model findings, distribution to both brain hemispheres is not likely following infusion into a single vertebral artery even if the infusate is well-mixed at the infusion site. This outcome results from the unique fluid flow properties of two converging channels, which are represented by the left and right vertebral branches converging into the basilar. Fluid in the model remains stratified on the side of the basilar artery served by the infused vertebral artery. Careful thought and planning of the methods of intravertebral drug infusions for treating posterior fossa tumors are required to assure proper distribution of the drug to the desired tissue regions. Improper delivery may be responsible for some noted toxicities or for failure of the treatments. C1 NIH, Div Bioeng & Phys Sci, Bethesda, MD 20892 USA. NIH, Neuro Oncol Branch, Bethesda, MD 20892 USA. NIH, Pediat Branch, Bethesda, MD 20892 USA. NIH, Dept Radiol, Bethesda, MD 20892 USA. RP Lutz, RJ (reprint author), NIH, Div Bioeng & Phys Sci, Bldg 13,Room 3N17, Bethesda, MD 20892 USA. NR 18 TC 6 Z9 6 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0167-594X J9 J NEURO-ONCOL JI J. Neuro-Oncol. PD JUN PY 2002 VL 58 IS 2 BP 95 EP 106 DI 10.1023/A:1016034910875 PG 12 WC Oncology; Clinical Neurology SC Oncology; Neurosciences & Neurology GA 581PN UT WOS:000177302200001 PM 12164691 ER PT J AU Cadet, JL AF Cadet, JL TI Involvement of multiple molecular events in methamphetamine-induced neurodegeneration: evidence from cDNA array analysis SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIDA, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 4 EP 4 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500014 ER PT J AU Romm, E Kim, JG Kim, NW Nagle, J Hudson, LD AF Romm, E Kim, JG Kim, NW Nagle, J Hudson, LD TI The MyT1 family recruits histone deacetylase to regulate neural transcription SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NINDS, NIH, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 5 EP 5 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500018 ER PT J AU Murphy, EJ Cole, N Glynn, A Nussbaum, RL AF Murphy, EJ Cole, N Glynn, A Nussbaum, RL TI Brain phospholipid levels and composition are altered in alpha-synuclen gene-abated mice SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Univ N Dakota, Grand Forks, ND 58201 USA. NHGRI, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 11 EP 11 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500039 ER PT J AU Rosenberger, TA Hovda, JT Peters, JM AF Rosenberger, TA Hovda, JT Peters, JM TI Disruption of PPAR beta results in distinct gender differences in mouse brain phospholipid SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. Penn State Univ, Ctr Mol Toxicol & Carcinogenesis, University Pk, PA 16802 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 13 EP 13 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500045 ER PT J AU Quarles, RH Dashiell, SD Weiss, MD Pant, HC AF Quarles, RH Dashiell, SD Weiss, MD Pant, HC TI Schwann cell to axon signalling mediated by myelin-associated glycoprotein affects the structure of myelinated axons SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NINDS, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 17 EP 17 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500058 ER PT J AU Marini, AM Banaudha, K McCall, S Zhu, D Lipsky, R AF Marini, AM Banaudha, K McCall, S Zhu, D Lipsky, R TI alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-mediated neuroprotection requires TRKB receptor activation SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Uniformed Serv Univ Hlth Sci, Dept Neurol, Bethesda, MD 20814 USA. USAF, Inst Pathol, Dept Cellular Pathol & Genet, Washington, DC 20330 USA. NIAAA, Neurogenet Lab, NIH, Rockville, MD 20852 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 26 EP 26 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500086 ER PT J AU Rosenberger, TA Rapoport, SI AF Rosenberger, TA Rapoport, SI TI Phospholipases A2, arachidonate turnover, and the neuroinflammatory response SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 31 EP 31 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500100 ER PT J AU Proia, RL AF Proia, RL TI Life without gangliosides SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIDDK, NIH, Bethesda, MD 20832 USA. RI Proia, Richard/A-7908-2012 NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 34 EP 34 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500112 ER PT J AU Mattson, MP Duan, W Lee, J Maswood, N Cheng, A AF Mattson, MP Duan, W Lee, J Maswood, N Cheng, A TI Dietary restriction, BDNF signalling and brain aging SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, Gerontol Res Ctr, Neurosci Lab, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 37 EP 37 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500122 ER PT J AU Wolkow, CA AF Wolkow, CA TI Insulin-like signalling in neurons controls lifespan in C-elegans SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, IRP, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 37 EP 37 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500119 ER PT J AU Barker, JL Maric, D AF Barker, JL Maric, D TI Transmitter signalling during early neurogenesis in the embryonic rat cortex SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NINDS, Neurophysiol Lab, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 38 EP 38 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500126 ER PT J AU Muja, N Lovas, G Romm, E Machleder, D Ranjan, M Hudson, LD AF Muja, N Lovas, G Romm, E Machleder, D Ranjan, M Hudson, LD TI Transgenic mice exhibiting oligodendrocyte-specific expression of a mutant protein tyrosine phosphatase epsilon SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NINDS, NIH, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 46 EP 46 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500142 ER PT J AU Nielsen, JA Hudson, LD Armstrong, RC AF Nielsen, JA Hudson, LD Armstrong, RC TI Myelin transcription factor 1 function in oligodendrocyte development SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 47 EP 47 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500146 ER PT J AU Lee, J Seroogy, KB Mattson, MP AF Lee, J Seroogy, KB Mattson, MP TI Dietary restriction enhances neurogenesis and up-regulates neurotrophin expression in the hippocampus of adult mice SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, LNS, GRC, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 0 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 57 EP 57 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500181 ER PT J AU Mattson, MP Cutler, RG Pedersen, WA AF Mattson, MP Cutler, RG Pedersen, WA TI Perturbed sphingomyelin metabolism and accumulation of ceramides and cholesterol esters in Alzheimer's disease and ALS SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 0 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 60 EP 60 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500190 ER PT J AU Wine, RN McPherson, CA Harry, GJ AF Wine, RN McPherson, CA Harry, GJ TI Elevation of cyclin D1 following trimethyltin induced hippocampal neurodegeneration SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 78 EP 78 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500247 ER PT J AU Martin, R AF Martin, R TI Use of pharmacogenomics in clinical trials for multiple sclerosis SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 81 EP 81 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500257 ER PT J AU Busciglio, J Pigino, G Morfini, G Mattson, MP Brady, ST AF Busciglio, J Pigino, G Morfini, G Mattson, MP Brady, ST TI Novel pathological mechanisms involving Presenilin 1 mutations SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Univ Connecticut, Ctr Hlth, Dept Neurosci, Farmington, CT USA. Univ Texas, SW Med Ctr, Dept Cell Biol, Dallas, TX USA. NIA, Neurosci Lab, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 0 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 84 EP 84 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500268 ER PT J AU Shine, HD Zhou, L Grider, MH McGowen, LR Menesses-Diaz, DA Zheng, X Le, WD Mamounas, L AF Shine, HD Zhou, L Grider, MH McGowen, LR Menesses-Diaz, DA Zheng, X Le, WD Mamounas, L TI Neurotrophic factor gene delivery supports axonal regeneration after injury SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Baylor Coll Med, Houston, TX 77030 USA. NINDS, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 86 EP 86 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500273 ER PT J AU Brunssen, SH Houser, ER Harry, GJ AF Brunssen, SH Houser, ER Harry, GJ TI Effects of early post-natal exposure of cerebral cortex to hyper-IL-6 on development and cognition in the CD-1 mouse SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIEHS, NIH, Res Triangle Pk, NC 27709 USA. UNC, Chapel Hill, NC USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 90 EP 90 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500279 ER PT J AU Grider, MH Le, WD Mamounas, LA Baumgartner, BJ Zheng, X Shine, HD AF Grider, MH Le, WD Mamounas, LA Baumgartner, BJ Zheng, X Shine, HD TI Adenoviral-mediated expression of BDNF induces axonal regeneration after neurotoxic lesion SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Baylor Coll Med, Houston, TX 77030 USA. Trinity Valley Community Coll, Athens, TX USA. NINDS, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 92 EP 92 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500285 ER PT J AU d'Hellencourt, CL Harry, GJ AF d'Hellencourt, CL Harry, GJ TI Injury-induced differential gene expression in laser captured hippocampal pyramidal and dentate granule cells SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 NIEHS, LMT, NIH, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 103 EP 103 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500320 ER PT J AU McPherson, CA Wine, RN Kubik, JA Harry, GJ AF McPherson, CA Wine, RN Kubik, JA Harry, GJ TI Trimethyltin induced hippocampal neurodegeneration elevates cyclins A and B SO JOURNAL OF NEUROCHEMISTRY LA English DT Meeting Abstract C1 Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 SU 1 BP 105 EP 105 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 573KU UT WOS:000176829500327 ER PT J AU Zapata, A Shippenberg, TS AF Zapata, A Shippenberg, TS TI D-3 receptor ligands modulate extracellular dopamine clearance in the nucleus accumbens SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE D-2 receptors; microdialysis; no-net flux; rotating disk electrode voltammetry; uptake ID IN-VIVO; RAT-BRAIN; QUANTITATIVE MICRODIALYSIS; STRIATAL TRANSPORTER; RELEASE; MICE; COCAINE; NEURONS; BINDING; AUTORECEPTORS AB An involvement of the D-3 dopamine receptor in the regulation of extracellular dopamine has been suggested. However, the mechanisms mediating this effect are unclear. We have used the technique of no net flux microdialysis under transient conditions to examine the influence of the D-3-preferring agonist (+)-PD128907 upon extracellular dopamine levels in the nucleus accumbens of the mouse. (+)-PD 128907 (0.1 mg/kg intraperitoneally) significantly decreased extracellular dopamine. This decrease was associated with a marked increase in the extraction fraction, which suggests an increase in dopamine clearance. The ability of D-3 -preferring compounds to modulate dopamine uptake was investigated in vitro using rotating disk electrode voltammetry. (+)-PD 128907 (10 nm) significantly increased the initial clearance rate of 3 mum dopamine in rat nucleus accumbens tissue suspensions. Kinetic analysis revealed no change in the apparent K-m of uptake but it showed a 33% increase in V-max. In contrast, the D-3 antagonist GR 103691 (10 nm) significantly decreased dopamine uptake. Consistent with the low levels of D-3 receptors in the dorsal striatum, neither compound affected uptake in tissue suspensions from this brain region. These data indicate that D-3 receptor activation increases dopamine uptake in the nucleus accumbens and suggest that this receptor subtype can regulate extracellular dopamine by modulating the DA transporter activity. C1 NIDA, Integrat Neurosci Sect, Behab Neurosci Branch, Intramural Res Program, Baltimore, MD 21224 USA. RP Zapata, A (reprint author), NIDA, Integrat Neurosci Sect, Behab Neurosci Branch, Intramural Res Program, 5500 Nathan Schock Dr, Baltimore, MD 21224 USA. NR 31 TC 40 Z9 40 U1 0 U2 2 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 IS 5 BP 1035 EP 1042 DI 10.1046/j.1471-4159.2002.00893.x PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 554HG UT WOS:000175729300015 PM 12065616 ER PT J AU Dashiell, SM Tanner, SL Pant, HC Quarles, RH AF Dashiell, SM Tanner, SL Pant, HC Quarles, RH TI Myelin-associated glycoprotein modulates expression and phosphorylation of neuronal cytoskeletal elements and their associated kinases SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE cyclin-dependent kinase-5; cytoskeleton; extracellular signal-regulated kinase; microtubule-associated proteins; myelin-associated glycoprotein; neurofilaments ID TANDEM MASS-SPECTROMETRY; SCHWANN-CELLS; NEUROFILAMENT PROTEIN; MICE DEFICIENT; SIALIC-ACID; MICROTUBULE STABILITY; PERIPHERAL-NERVES; AXONAL-TRANSPORT; TREMBLER MICE; TAIL DOMAIN AB Decreased phosphorylation of neurofilaments in mice lacking myelin-associated glycoprotein (MAG) was shown to be associated with decreased activities of extracellular-signal regulated kinases (ERK1/2) and cyclin-dependent kinase-5 (cdk5). These in vivo changes could be caused directly by the absence of a MAG-mediated signaling pathway or secondary to a general disruption of the Schwann cell-axon junction that prevents signaling by other molecules. Therefore, in vitro experimental paradigms of MAG interaction with neurons were used to determine if MAG directly influences expression and phosphorylation of cytoskeletal proteins and their associated kinases. COS-7 cells stably transfected with MAG or with empty vector were co-cultured with primary dorsal root ganglion (DRG) neurons. Total amounts of the middle molecular weight neurofilament subunit (NF-M), microtubule-associated protein 1B (MAP1B), MAP2, and tau were up-regulated significantly in DRG neurons in the presence of MAG. There was also increased expression of phosphorylated high molecular weight neurofilament subunit (NF-H), NF-M, and MAP1B. Additionally, in similar in vitro paradigms, total and phosphorylated NF-M were increased significantly in PC12 neurons co-cultured with MAG-expressing COS cells or treated with a soluble MAG Fc-chimera. The increased expression of phosphorylated cytoskeletal proteins in the presence of MAG in vitro was associated with increased activities of ERK 1/2 and cdk5. We propose that interaction of MAG with an axonal receptor(s) induces a signal transduction cascade that regulates expression of cytoskeletal proteins and their phosphorylation by these proline-directed protein kinases. C1 NINCDS, Mol & Cellular Neurobiol Lab, NIH, Bethesda, MD 20892 USA. NINCDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Quarles, RH (reprint author), NINCDS, Mol & Cellular Neurobiol Lab, NIH, Room 2A28,Bldg 49,MSC 4440,49 Convent Dr, Bethesda, MD 20892 USA. NR 56 TC 70 Z9 73 U1 0 U2 3 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 IS 6 BP 1263 EP 1272 DI 10.1046/j.1471-4159.2002.00927.x PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 559ZN UT WOS:000176056700012 PM 12068074 ER PT J AU Gao, HM Jiang, J Wilson, B Zhang, W Hong, JS Liu, B AF Gao, HM Jiang, J Wilson, B Zhang, W Hong, JS Liu, B TI Microglial activation-mediated delayed and progressive degeneration of rat nigral dopaminergic neurons: relevance to Parkinson's disease SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE dopamine; inflammation; microglia; Parkinson's disease; superoxide ID NECROSIS-FACTOR-ALPHA; ALZHEIMERS-DISEASE; NITRIC-OXIDE; INTRANIGRAL INJECTION; SUBSTANTIA-NIGRA; BRAIN INJURY; INFLAMMATORY DAMAGE; NADPH OXIDASE; GLIAL-CELLS; LIPOPOLYSACCHARIDE AB The etiology of sporadic Parkinson's disease (PD) remains unknown. Increasing evidence has suggested a role for inflammation in the brain in the pathogenesis of PD. However, it has not been clearly demonstrated whether microglial activation, the most integral part of the brain inflammatory process, will result in a delayed and progressive degeneration of dopaminergic neurons in substantia nigra, a hallmark of PD. We report here that chronic infusion of an inflammagen lipopolysaccharide at 5 ng/h for 2 weeks into rat brain triggered a rapid activation of microglia that reached a plateau in 2 weeks, followed by a delayed and gradual loss of nigral dopaminergic neurons that began at between 4 and 6 weeks and reached 70% by 10 weeks. Further investigation of the underlying mechanism of action of microglia-mediated neurotoxicity using rat mesencephalic neuron-glia cultures demonstrated that low concentrations of lipopolysaccharide (0.1-10 ng/mL)-induced microglial activation and production of neurotoxic factors preceded the progressive and selective degeneration of dopaminergic neurons. Among the factors produced by activated microglia, the NADPH oxidase-mediated release of superoxide appeared to be a predominant effector of neurodegeneration, consistent with the notion that dopaminergic neurons are particularly vulnerable to oxidative insults. This is the first report that microglial activation induced by chronic exposure to inflammagen was capable of inducing a delayed and selective degeneration of nigral dopaminergic neurons and that microglia-originated free radicals play a pivotal role in dopaminergic neurotoxicity in this inflammation-mediated model of PD. C1 NIEHS, Neuropharmacol Sect, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC USA. Dalian Med Univ, Dept Physiol, Dalian, Peoples R China. RP Liu, B (reprint author), F1-01,POB 12233, Res Triangle Pk, NC 27709 USA. RI gao, huiming/C-8454-2012; liu, Bin/A-7695-2009 NR 53 TC 401 Z9 440 U1 5 U2 23 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 IS 6 BP 1285 EP 1297 DI 10.1046/j.1471-4159.2002.00928.x PG 13 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 559ZN UT WOS:000176056700014 PM 12068076 ER PT J AU Lee, HK Safieddine, S Petralia, RS Wenthold, RJ AF Lee, HK Safieddine, S Petralia, RS Wenthold, RJ TI Identification of a novel SNAP25 interacting protein (SIP30) SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE SNAP25; SNAREs; syntaxin; yeast two-hybrid ID HAIR-CELLS; INFERIOR COLLICULUS; NEUROTRANSMITTER RELEASE; SYNAPTIC VESICLES; SNARE COMPLEXES; ADULT CAT; EXOCYTOSIS; SYNTAXIN; SNAP-25; ORGANIZATION AB Soluble N -ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), including synaptosome-associated proteins of 25 kDa (SNAP25), syntaxins, and vesicle-associated membrane proteins (VAMP), are essential for regulated exocytosis of synaptic vesicles in neurotransmission. We identified a cDNA coding for a novel protein of 266 amino acids that we have named SIP30 (S NAP25 interacting protein of 30 kDa). SIP30 is expressed abundantly in brain and slightly in testis and kidney. In brain, SIP30 is highly expressed in the inferior and superior colliculi, which contain important relay nuclei of the auditory and visual systems. GST-pull-down and immunoprecipitation assays showed direct binding of SIP30 to SNAP25. Although SIP30 does not directly interact with syntaxin based on pull-down assays, syntaxin does co-immunoprecipitate with SIP30 suggesting that syntaxin is indirectly associated with SIP30, perhaps through SNAP25. C1 NIDCD, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Wenthold, RJ (reprint author), NIDCD, Neurochem Lab, NIH, Bldg 50,Room 4140, Bethesda, MD 20892 USA. NR 39 TC 15 Z9 16 U1 0 U2 0 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUN PY 2002 VL 81 IS 6 BP 1338 EP 1347 DI 10.1046/j.1471-4159.2002.00937.x PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 559ZN UT WOS:000176056700019 PM 12068081 ER PT J AU Devanne, H Cohen, LG Kouchtir-Devanne, N Capaday, C AF Devanne, H Cohen, LG Kouchtir-Devanne, N Capaday, C TI Integrated motor cortical control of task-related muscles during pointing in humans SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID TRANSCRANIAL MAGNETIC STIMULATION; SOLEUS H-REFLEX; CORTICOCORTICAL INHIBITION; INTRACORTICAL INHIBITION; HUMAN WALKING; CORTEX; FORELIMB; INPUT; CAT; REPRESENTATIONS AB A large body of compelling but indirect evidence suggests that the motor cortex controls the different forelimb segments as a whole rather than individually. The purpose of this study was to obtain physiological evidence in behaving human subjects on the mode of operation of the primary motor cortex during coordinated movements of the forelimb. We approached this problem by studying a pointing movement involving the shoulder, elbow, wrist, and index finger as follows. Focal transcranial magnetic stimulation (TMS) was used to measure the input-output (I/O) curves-a measure of the corticospinal pathway excitability-of proximal (anterior deltoid, AD, and triceps brachii, TB) and distal muscles (extensor carpi radialis, ECR, and first dorsal interosseus, 1DI) during isolated contraction of one of these muscles or during selective co-activation with other muscles involved in pointing. Compared to an isolated contraction of the ECR, the plateau-level of the ECR sigmoid I/O curve increased markedly during co-activation with the AD while pointing. In contrast, the I/O curve of AD was not influenced by activation of the more distal muscles involved in pointing. Moreover, the 1DI I/O curve was not influenced by activation of the more proximal muscles. Three arguments argue for a cortical site of facilitation of ECR motor potentials. First, ECR motor potentials evoked by a near threshold TMS stimulus were facilitated when the AD and ECR were co-activated during pointing but not those in response to a near threshold anodal electrical stimulus. Second, the ECR H reflex was not found to be task dependent, indicating that the recruitment gain of the ECR alpha-motoneuron pool did not differ between tasks. Finally, in comparison with an isolated ECR contraction, intracortical inhibition tested at the ECR cortical site was decreased during pointing. These results suggest that activation of shoulder, elbow, and wrist muscles involved in pointing appear to involve, at least in part, common motor cortical circuits. In contrast, at least in the pointing task, the motor cortical circuits involved in activation of the 1DI appear to act independently. C1 Univ Laval Robert Giffard, Ctr Rech, Brain & Movement Lab, Dept Anat & Physiol, Quebec City, PQ G1J 2G3, Canada. Ctr Hosp Reg & Univ Lille, F-59037 Lille, France. Univ Littoral Cote Opale, F-62228 Calais, France. NINCDS, Human Cort Physiol Sect, NIH, Bethesda, MD 20982 USA. CNRS, FRE, Lab Neurophys & Physiol Syst Moteur, F-75006 Paris, France. RP Capaday, C (reprint author), Univ Laval Robert Giffard, Ctr Rech, Brain & Movement Lab, Dept Anat & Physiol, F-6500,2601 Canardiere, Quebec City, PQ G1J 2G3, Canada. RI Devanne, Herve/M-7703-2016 OI Devanne, Herve/0000-0002-0296-9152 NR 40 TC 57 Z9 57 U1 0 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD JUN PY 2002 VL 87 IS 6 BP 3006 EP 3017 DI 10.1152/jn.00990.2001 PG 12 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 556ZA UT WOS:000175878900039 PM 12037204 ER PT J AU Bichot, NP Schall, JD AF Bichot, NP Schall, JD TI Priming in macaque frontal cortex during popout visual search: Feature-based facilitation and location-based inhibition of return SO JOURNAL OF NEUROSCIENCE LA English DT Article DE visual cortex; vision; attention; selection; eye movements; oculomotor ID EYE FIELD NEURONS; ALTERNATIVE FORCED-CHOICE; SACCADE TARGET SELECTION; INFERIOR TEMPORAL CORTEX; NEURAL MECHANISMS; PREFRONTAL CORTEX; VISUOSPATIAL ATTENTION; SINGLE NEURONS; REACTION-TIME; MONKEY AB In popout search, humans and monkeys are affected by trial-to-trial changes in stimulus features and target location. The neuronal mechanisms underlying such sequential effects have not been examined. Single neurons were recorded in the frontal eye field (FEF) of monkeys performing a popout search during which stimulus features and target position changed unpredictably across trials. Like previous studies, repetition of stimulus features improved performance. This feature-based facilitation of return was manifested in the target discrimination process in FEF: neurons discriminated the target from distractors earlier and better with repetition of stimulus features, corresponding to improvements in saccade latency and accuracy, respectively. The neuronal target selection was mediated by both target enhancement and distractor suppression. In contrast to the repetition of features, repetition of target position increased saccade latency. This location-based inhibition of return was reflected in the neuronal discrimination process but not in the baseline activity in FEF. These results show adjustments of the target selection process in FEF corresponding to and therefore possibly contributing to changes in performance across trials caused by sequential regularities in display properties. C1 NIMH, Neuropsychol Lab, NIH, Bethesda, MD 20892 USA. Vanderbilt Univ, Dept Psychol, Vanderbilt Vis Res Ctr, Nashville, TN 37240 USA. RP Bichot, NP (reprint author), NIMH, Neuropsychol Lab, NIH, Bldg 49,Room 1B80, Bethesda, MD 20892 USA. FU NEI NIH HHS [P30-EY08126, R01-EY08890, T32-EY0135] NR 71 TC 145 Z9 148 U1 0 U2 7 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 JUN 1 PY 2002 VL 22 IS 11 BP 4675 EP 4685 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 557GL UT WOS:000175900700048 PM 12040074 ER PT J AU Berti, R Brennan, MB Soldan, SS Ohayon, JM Casareto, L McFarland, HF Jacobson, S AF Berti, R Brennan, MB Soldan, SS Ohayon, JM Casareto, L McFarland, HF Jacobson, S TI Increased detection of serum HHV-6 DNA sequences during multiple sclerosis (MS) exacerbations and correlation with parameters of MS disease progression SO JOURNAL OF NEUROVIROLOGY LA English DT Article DE HHV-6; multiple sclerosis; exacerbation ID POLYMERASE-CHAIN-REACTION; HUMAN HERPESVIRUS-6; NERVOUS-SYSTEM; INFECTION; ASSOCIATION; PATHOGEN; CHILDREN; VARIANT AB In recent years, human herpesvirus 6 (HHV-6) has been investigated as a possible causative agent for MS. To determine if the detection of HHV-6 DNA in the serum of MS patients correlates with clinical parameters of MS disease progression, a total of 215 serum samples was obtained from 59 MS patients followed prospectively for a 5-month period. These samples were analyzed for the presence of HHV-6 DNA by nested PCR and compared in parallel to MS disease activity. HHV-6 DNA was amplified in 22% (4/18) of samples obtained during a period of clinical exacerbation. Significantly fewer (P = 0.008) sera, 5.6% (11/197), obtained from MS patients during clinical remission tested positive for the presence of HHV-6 DNA. This work demonstrates that the detection of serum HHV-6 DNA is significantly correlated with clinical exacerbations in MS. Moreover, the findings presented in this study have confirmed previous reports supporting an association between MS and HHV-6 and suggest a role for this human herpesvirus in the pathogenesis of MS. C1 NINDS, Viral Immunol Sect, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Walter Reed Army Inst Res, Dept Neuropharmacol & Mol Biol, Silver Spring, MD USA. George Washington Univ, Dept Genet, Inst Biomed Sci, Washington, DC 20052 USA. InfoMax Inc, Bethesda, MD USA. RP Jacobson, S (reprint author), NINDS, Viral Immunol Sect, Neuroimmunol Branch, NIH, Bldg 10,Room 5B16, Bethesda, MD 20892 USA. NR 34 TC 52 Z9 52 U1 0 U2 1 PU TAYLOR & FRANCIS INC PI PHILADELPHIA PA 325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USA SN 1355-0284 J9 J NEUROVIROL JI J. Neurovirol. PD JUN PY 2002 VL 8 IS 3 BP 250 EP 256 DI 10.1080/13550280290049615 PG 7 WC Neurosciences; Virology SC Neurosciences & Neurology; Virology GA 570QR UT WOS:000176669500010 PM 12053279 ER PT J AU Mariani, G Gipponi, M Moresco, L Villa, G Bartolomei, M Mazzarol, G Bagnara, MC Romanini, A Cafiero, F Paganelli, G Strauss, HW AF Mariani, G Gipponi, M Moresco, L Villa, G Bartolomei, M Mazzarol, G Bagnara, MC Romanini, A Cafiero, F Paganelli, G Strauss, HW TI Radioguided sentinel lymph node biopsy in malignant cutaneous melanoma SO JOURNAL OF NUCLEAR MEDICINE LA English DT Review DE sentinel lymph node; malignant cutaneous melanoma; radiocolloid; interstitial administration; lymphoscintigraphy; intraoperative gamma-probe guidance ID POLYMERASE-CHAIN-REACTION; EARLY-STAGE MELANOMA; INTRAOPERATIVE GAMMA-PROBE; TYROSINASE MESSENGER-RNA; AMERICAN JOINT COMMITTEE; HIGH-RISK MELANOMA; BREAST-CANCER; RADIATION-PROTECTION; DYNAMIC LYMPHOSCINTIGRAPHY; DOSE INTERFERON-ALPHA-2B AB The procedure of sentinel lymph node biopsy in patients with malignant cutaneous melanoma has evolved from the notion that the tumor drains in a logical way through the lymphatic system, from the first to subsequent levels. As a consequence, the first lymph node encountered (the sentinel node) will most likely be the first affected by metastasis; therefore, a negative sentinel node makes it highly unlikely that other nodes in the same lymphatic basin are affected. Although the long-term therapeutic benefit of the sentinel lymph node biopsy per se has not yet been ascertained, this procedure distinguishes patients without nodal metastases, who can avoid nodal basin dissection with its associated risk of lymphedema, from those with metastatic involvement, who may benefit from additional therapy. Sentinel lymph node biopsy would represent a significant advantage as a minimally invasive procedure, considering that an average of only 20% of melanoma patients with a Breslow thickness between 1.5 and 4 mm harbor metastasis in their sentinel node and are therefore candidates for elective lymph node dissection. Furthermore, histologic sampling errors (amounting to approximately 12% of lymph nodes in the conventional routine) can be reduced if one assesses a single (sentinel) node extensively rather than assessing the standard few histologic sections in a high number of lymph nodes per patient. The cells from which cutaneous melanomas originate are located between the dermis and the epidermis, a zone that drains to the inner lymphatic network in the reticular dermis and, in turn, to larger collecting lymphatics in the subcutis. Therefore, the optimal route for interstitial administration of radiocolloids for lymphoscintigraphy and subsequent radioguided sentinel lymph node biopsy is intradermal or subdermal injection. Tc-99m-Labeled colloids in various size ranges are equally adequate for radioguided sentinel lymph node biopsy in patients with cutaneous melanoma, depending on local experience and availability. For melanomas along the midline of the head, neck, and trunk, particular consideration should be given to ambiguous lymphatic drainage, which frequently requires interstitial administration virtually all around the tumor or surgical scar from prior excision of the melanoma. Lymphoscintigraphy is an essential part of radioguided sentinel lymph node biopsy because images are used to direct the surgeon to the sites of the nodes. The sentinel lymph node should have a significantly higher count than that of the background (at least 10:1 intraoperatively). After removal of the sentinel node, the surgical bed must be reexamined to ensure that all radioactive sites are identified and removed for analysis. Virtually the entire sentinel lymph node should be processed for histopathology, including both conventional hematoxylin-eosin staining and immune staining with antibodies to the S-100 and HMB-45 antigens. The success rate of radioguidance in localizing the sentinel lymph node in melanoma patients is approximately 98% in institutions that perform a high number of procedures and approaches 99% when combined with the vital blue-dye technique. Growing evidence of the high correlation between a sentinel lymph node biopsy negative for cancer and a negative status for the lymphatic basin-evidence, therefore, of the high prognostic value of sentinel node biopsy-has led to the procedure's being included in the most recent version of the TNM staging system and starting to become the standard of care for patients with cutaneous melanoma. C1 Univ Pisa, Sch Med, Reg Ctr Nucl Med, I-56126 Pisa, Italy. CNR, Inst Clin Physiol, Pisa, Italy. Natl Canc Inst, Div Surg Oncol, Genoa, Italy. Univ Genoa, Dept Oncol Biol & Genet, Genoa, Italy. Univ Genoa, Sch Med, Dept Internal Med & Med Specialities, Nucl Med Serv, Genoa, Italy. European Inst Oncol, Div Nucl Med, Milan, Italy. European Inst Oncol, Dept Pathol, Milan, Italy. Osped San Martino Genova, Hlth Phys Unit, Genoa, Italy. Santa Chiara Hosp, Med Oncol Unit, Pisa, Italy. Mem Sloan Kettering Canc Ctr, Dept Radiol, Div Nucl Med, New York, NY USA. RP Mariani, G (reprint author), Univ Pisa, Sch Med, Reg Ctr Nucl Med, Via Roma 67, I-56126 Pisa, Italy. NR 157 TC 79 Z9 81 U1 1 U2 1 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD JUN PY 2002 VL 43 IS 6 BP 811 EP 827 PG 17 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 559AJ UT WOS:000176001700021 PM 12050328 ER PT J AU McKeown, NM Jacques, PF Gundberg, CM Peterson, JW Tucker, KL Kiel, DP Wilson, PWF Booth, SL AF McKeown, NM Jacques, PF Gundberg, CM Peterson, JW Tucker, KL Kiel, DP Wilson, PWF Booth, SL TI Dietary and nondietary determinants of vitamin K biochemical measures in men and women SO JOURNAL OF NUTRITION LA English DT Article DE dietary methods; biomarker; vitamin K; osteocalcin; Framingham Offspring Study ID FOOD FREQUENCY QUESTIONNAIRE; FAT-SOLUBLE VITAMINS; PHYLLOQUINONE VITAMIN-K-1; SERUM CONCENTRATIONS; BONE HEALTH; PLASMA; OSTEOCALCIN; ADULTS; REPRODUCIBILITY; OSTEOPOROSIS AB Few epidemiological studies that rely on the food frequency questionnaire (FFQ) for dietary assessment have measured biomarkers of vitamin K intake to independently confirm associations between self-reported dietary vitamin K intake and disease risk. Associations were examined between two sensitive biomarkers of vitamin K status, plasma phylloquinone and serum percent undercarboxylated osteocalcin (%ucOC), and self-reported usual phylloquinone intake as estimated from a FFQ. The influence of other dietary and nondietary factors on plasma phylloquinone concentrations was also examined. Dietary phylloquinone intake was estimated using a FFQ in 369 men and 468 women of the Framingham Offspring Study. The prevalence of high %ucOC concentrations (greater than or equal to 20%), suggestive of a low vitamin K status, was 44% in men and 54% in women, respectively. After multivariate adjustment, the odds of a high %ucOC was 2.5 greater for women (odds ratio: 2.5; 95% confidence interval [CI]: 1.2-5.1) and almost three times greater for men (odds ratio: 2.8; 95% CI: 1.3-5.9) in the lowest dietary phylloquinone intake quintile category compared to the highest quintile category. Fasting triglyceride concentrations, smoking status and season were associated with plasma phylloquinone concentrations, independent of dietary phylloquinone intake. Phylloquinone and green vegetable intake was linearly associated with plasma phylloquinone, after adjustment for potential confounding factors. There were limitations in the use of the FFQ to predict plasma phylloquinone, evident in an observed plateau effect and required nondietary adjustment factors. Despite these caveats, these findings support the use of a FFQ for a relative assessment of vitamin K status in population-based studies. C1 Tufts Univ, Jena Mayer US Dept Agr Human Nutr Res Ctr Aging, Boston, MA 02111 USA. Yale Univ, Sch Med, New Haven, CT 06510 USA. Hebrew Rehabil Ctr Aged, Res & Training Inst, Boston, MA 02131 USA. Harvard Univ, Sch Med, Boston, MA 02131 USA. Boston Univ, Sch Med, Boston, MA 01702 USA. NHLBI, Framingham Heart Study, Boston, MA 01702 USA. RP Booth, SL (reprint author), Tufts Univ, Jena Mayer US Dept Agr Human Nutr Res Ctr Aging, Boston, MA 02111 USA. RI Tucker, Katherine/A-4545-2010; OI Kiel, Douglas/0000-0001-8474-0310; Tucker, Katherine/0000-0001-7640-662X FU NHLBI NIH HHS [N01-HC-38038]; NIA NIH HHS [AG 14759]; NIDDK NIH HHS [T32 DK 07651] NR 36 TC 80 Z9 82 U1 1 U2 3 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD JUN PY 2002 VL 132 IS 6 BP 1329 EP 1334 PG 6 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 559NY UT WOS:000176033300038 PM 12042454 ER PT J AU Tewari, KS DiSaia, PJ AF Tewari, KS DiSaia, PJ TI Radiation therapy for gynecologic cancer SO JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH LA English DT Article ID CERVICAL-CANCER; INTERSTITIAL BRACHYTHERAPY; BREAST-CANCER; ENDOMETRIAL CANCER; PROGNOSTIC FACTORS; OVARIAN-CANCER; DOSE-RATE; CARCINOMA; RADIOTHERAPY; VULVA C1 Univ Calif Irvine, Div Gynecol Oncol, Orange, CA 92868 USA. RP DiSaia, PJ (reprint author), Chao Family NCI Designated Comprehens Canc Ctr, 101 City Dr, Orange, CA 92868 USA. NR 91 TC 5 Z9 5 U1 0 U2 0 PU BLACKWELL PUBLISHING ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 1341-8076 J9 J OBSTET GYNAECOL RE JI J. Obstet. Gynaecol. Res. PD JUN PY 2002 VL 28 IS 3 BP 123 EP 140 DI 10.1046/j.1341-8076.2002.00036.x PG 18 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 585HZ UT WOS:000177520500001 PM 12214828 ER PT J AU Packer, RJ Biegel, JA Blaney, S Finlay, J Geyer, JR Heideman, R Hilden, J Janss, AJ Kun, L Vezina, G Rorke, LB Smith, M AF Packer, RJ Biegel, JA Blaney, S Finlay, J Geyer, JR Heideman, R Hilden, J Janss, AJ Kun, L Vezina, G Rorke, LB Smith, M TI Atypical teratoid/rhabdoid tumor of the central nervous system: Report on workshop SO JOURNAL OF PEDIATRIC HEMATOLOGY ONCOLOGY LA English DT Article DE atypical teratoid tumors; rhabdoid tumors; infantile brain tumors; radiotherapy; chemotherapy ID MALIGNANT RHABDOID TUMOR; TERATOID TUMOR; INFANCY; CHILDHOOD; MUTATIONS; ENTITY; DEFINITION; BRAIN AB Childhood atypical teratoid/rhabdoid tumor (AT/RT) of the central nervous system (CNS) is a recently described entity. Diagnosis is based on distinctive light microscopy and immunohistochemical findings, coupled with molecular genetic analysis. Most AT/RTs demonstrate monosomy 22 or deletions of chromosome band 22q11 with alterations of the hSNF5/INI1 gene. The tumor's incidence is still undefined, but it may comprise as big as 1 in 4 primitive CNS tumors in infants. Treatment is far from optimal, but there are occasional long-term survivors, especially among older children. Therapeutic approached have included surgery, chemotherapy, and radiotherapy.Prospective clinical trials are needed for children with AT/RTs. C1 George Washington Univ, Ctr Neurosci & Behav Med, Childrens Natl Med Ctr, Med Ctr,Dept Neurol, Washington, DC 20010 USA. George Washington Univ, Dept Pediat, Childrens Natl Med Ctr, Med Ctr, Washington, DC 20010 USA. George Washington Univ, Dept Neuroradiol, Childrens Natl Med Ctr, Med Ctr, Washington, DC 20010 USA. Univ Penn, Childrens Hosp, Div Human Genet, Philadelphia, PA 19104 USA. Univ Penn, Childrens Hosp, Div Pediat, Philadelphia, PA 19104 USA. Univ Penn, Childrens Hosp, Div Neuropathol, Philadelphia, PA 19104 USA. Texas Childrens Hosp, Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. NYU, Dept Pediat, New York, NY 10016 USA. Univ Washington, Childrens Reg Med Ctr, Dept Pediat, Seattle, WA 98195 USA. St Jude Childrens Res Hosp, Dept Pediat, Memphis, TN 38105 USA. St Jude Childrens Res Hosp, Dept Radiat Oncol, Memphis, TN 38105 USA. Cleveland Clin Fdn, Dept Pediat, Cleveland, OH 44195 USA. NCI, Bethesda, MD 20892 USA. RP Packer, RJ (reprint author), George Washington Univ, Ctr Neurosci & Behav Med, Childrens Natl Med Ctr, Med Ctr,Dept Neurol, 111 Michigan Ave NW, Washington, DC 20010 USA. EM rpacker@cnmc.org NR 15 TC 131 Z9 133 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1077-4114 J9 J PEDIAT HEMATOL ONC JI J. Pediatr. Hematol. Oncol. PD JUN-JUL PY 2002 VL 24 IS 5 BP 337 EP 342 DI 10.1097/00043426-200206000-00004 PG 6 WC Oncology; Hematology; Pediatrics SC Oncology; Hematology; Pediatrics GA 570ZR UT WOS:000176690800002 PM 12142780 ER PT J AU Uwaifo, GI Nguyen, TT Keil, MF Russell, DL Nicholson, JC Bonat, SH McDuffie, JR Yanovski, JA AF Uwaifo, GI Nguyen, TT Keil, MF Russell, DL Nicholson, JC Bonat, SH McDuffie, JR Yanovski, JA TI Differences in insulin secretion and sensitivity of Caucasian and African American prepubertal children SO JOURNAL OF PEDIATRICS LA English DT Article ID C-PEPTIDE CONCENTRATIONS; BETA-CELL FUNCTION; WHITE AMERICANS; HYPERGLYCEMIC CLAMP; HEPATIC EXTRACTION; BLACK; GLUCOSE; ADOLESCENTS; CLEARANCE; OBESITY AB Objective: To investigate whether differences in body composition of African American children (AA) and Caucasian children (C) explain differences in insulin sensitivity and secretion. Study design: Prepubertal nondiabetic children (31 AA and 54 C) were studied; 84% were overweight. Participants under-went a 2-hour hyperglycemic clamp, to estimate insulin sensitivity (SI (clamp)) and secretion, and dual energy x-ray absorptiometry, to assess body composition. Results: AA had greater total body fat mass (P = .01), fasting, 1st phase, 2nd phase, and steady state insulin levels (P < .05). AA and C had similar glucose disposal rates, but AA had lower SIclamp steady state C-peptide were less in C (P < .05), whereas corresponding C- peptide/insulin ratios were higher (all P < .005). Insulin levels and SIclamp remained different in AA and C after adjustment for body fat or lean mass differences. Analyses restricted to only overweight AA and C showed similar trends. Conclusion: Prepubertal African American children have higher baseline and glucose-stimulated insulin and C-peptide levels, as welt as reduced insulin sensitivity that is not entirely explained by differences in adiposity. The lower C-peptide/insulin molar ratio in AA suggests that they probably have lower hepatic insulin clearance than Caucasian children. C1 NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Uwaifo, GI (reprint author), NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, NIH, 10 Ctr Dr,Bldg 10,Rm 10N262,MSC 1862, Bethesda, MD 20892 USA. RI Uwaifo, Gabriel/M-2361-2016; OI Uwaifo, Gabriel/0000-0002-6962-9304; Yanovski, Jack/0000-0001-8542-1637 FU NICHD NIH HHS [HD-000641] NR 35 TC 28 Z9 29 U1 0 U2 0 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 JUN PY 2002 VL 140 IS 6 BP 673 EP 680 DI 10.1067/mpd.2002.124312 PG 8 WC Pediatrics SC Pediatrics GA 568AM UT WOS:000176520300008 PM 12072869 ER PT J AU Fox, E Bungay, PM Bacher, J McCully, CL Dedrick, RL Balis, FM AF Fox, E Bungay, PM Bacher, J McCully, CL Dedrick, RL Balis, FM TI Zidovudine concentration in brain extracellular fluid measured by microdialysis: Steady-state and transient results in rhesus monkey SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID CEREBROSPINAL-FLUID; CONTINUOUS INFUSION; PHARMACOKINETICS; CALIBRATION; TRANSPORT; INVITRO; INVIVO; PROBES; WATER AB We measured zidovudine concentrations in blood, muscle, and brain extracellular fluid (ECF) by microdialysis and in serum ultrafiltrate and cerebrospinal fluid (CSF) samples during a continuous intravenous infusion (15 mg/kg/h) and after bolus dosing (50-80 mg/kg over 15 min) in nonhuman primates to determine whether CSF drug penetration is a valid surrogate for blood-brain barrier penetration. Recovery was estimated in vivo by zero net flux for the continuous infusion and retrodialysis for the bolus dosing. In vivo recovery was tissue-dependent and was lower in brain than in blood or muscle. Mean (+/-S.D.) steady-state blood, muscle, and brain zidovudine concentrations by microdialysis were 112 +/- 63.8, 105 +/- 51.1, and 13.8 +/- 10.4 muM, respectively; and steady-state serum ultrafiltrate and CSF concentrations were 81.2 +/- 40.2 and 14.1 +/- 8.0 muM, respectively. Brain ECF penetration (microdialysis brain/blood ratio) and CSF penetration (standard sampling CSF/serum ratio) at steady state were 0.13 +/- 0.06 and 0.17 +/- 0.02, respectively. With bolus dosing the mean (+/-S.D.) zidovudine area under concentration-time curve (AUC) normalized to a dose of 80 mg/kg was 577 +/- 103 muM . h in blood, 528 +/- 202 muM . h in muscle, and 108 +/- 74 muM . h in brain (brain/blood ratio of 0.18 +/- 0.10) by microdialysis. Serum ultrafiltrate AUC was 446 +/- 72 muM . h and the CSF AUC was 123 +/- 4.7 muM . h (CSF/serum ratio of 0.28 +/- 0.06). In conclusion, recovery was tissue-dependent. CSF and brain ECF zidovudine concentrations were comparable at steady state, and the corresponding AUCs were comparable after bolus injection. Thus, zidovudine penetration in brain ECF and CSF in nonhuman primates is limited to a similar extent, presumably by active transport, as in other species. C1 NCI, Pharmacol & Expt Therapeut Sect, Pediat Oncol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NIH, Div Bioengn & Phys Sci, Bethesda, MD 20892 USA. NIH, Surg Serv, Vet Resources Program, Off Res Serv, Bethesda, MD 20892 USA. RP Fox, E (reprint author), NCI, Pharmacol & Expt Therapeut Sect, Pediat Oncol Branch, Ctr Canc Res, 10-13C103,10 Ctr Dr,MSC 1920, Bethesda, MD 20892 USA. NR 27 TC 26 Z9 27 U1 0 U2 1 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD JUN PY 2002 VL 301 IS 3 BP 1003 EP 1011 AR UNSP 0/986121 DI 10.1124/jpet.301.3.1003 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LK UT WOS:000175736800028 PM 12023531 ER PT J AU Granvil, CP Krausz, KW Gelboin, HV Idle, JR Gonzalez, FJ AF Granvil, CP Krausz, KW Gelboin, HV Idle, JR Gonzalez, FJ TI 4-hydroxylation of debrisoquine by human CYP1A1 and its inhibition by quinidine and quinine SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID HUMAN-LIVER-MICROSOMES; METABOLIZING ENZYME EXPRESSION; CYTOCHROME-P450 2D6; POOR METABOLIZERS; LIQUID-CHROMATOGRAPHY; MASS-SPECTROMETRY; HYDROXYLATION POLYMORPHISM; SUBSTRATE-SPECIFICITY; CYP2D6 POLYMORPHISM; LUNG-CANCER AB A panel of 15 recombinant cytochromes P450 expressed in human B-lymphoblastoid cells was used to study debrisoquine 4-hydroxylation. Both CYP2D6 and CYP1A1 carried out the reaction. The apparent K-m (micromolar) and V-max (picomoles per minute per picomole of P450) for CYP2D6 were 12.1 and 18.2 and for CYP1A1 were 23.1 and 15.2, respectively. CYP1A1 debrisoquine 4-hydroxylase was inhibited by the CYP1A1 inhibitor alpha-naphthoflavone and the CYP1A1 substrate 7-ethoxyresorufin. Additionally and surprisingly, this reaction was also inhibited by quinidine and quinine, with respective IC50 values of 1.38 +/- 0.10 and 3.31 +/- 0.14 muM, compared with those for CYP2D6 debrisoquine 4-hydroxylase of 0.018 +/- 0.05 and 3.75 +/- 2.07 muM, respectively. Anti-CYP1A1 monoclonal antibody (mAb) 1-7-1 abolished CYP1A1 debrisoquine hydroxylase and anti-CYP2D6 mAb 50-1-3 eradicated CYP2D6 debrisoquine 4-hydroxylase. Three further CYP2D6-specific reactions were tested: dextromethorphan O-demethylation, bufuralol 1'-hydroxylation, and sparteine dehydrogenation. The CYP2D6 specificity, judged by the CYP2D6/CYP1A1 activity ratios was 18.5, 7.0, 6.0, and 1.6 for dextromethorphan, bufuralol, sparteine, and debrisoquine, respectively. Thus, debrisoquine is not a specific CYP2D6 substrate and quinidine is not a specific CYP2D6 inhibitor. These findings have significant implications for the conduct of in vitro drug metabolism inhibition studies and underscore the fallacy of "specific chemical inhibitors" of a supergene family of enzymes that have overlapping substrate specificities. The use of highly specific mAbs in such studies is mandated. It is unclear as yet whether these findings have implications for the relationship between CYP2D6 genotype and in vivo debrisoquine 4-hydroxylase activity. C1 NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Norwegian Univ Sci & Technol, Inst Canc Res & Mol Biol, N-7034 Trondheim, Norway. RP Gonzalez, FJ (reprint author), NCI, Lab Metab, NIH, Bldg 37,Room 3E24, Bethesda, MD 20892 USA. OI Idle, Jeff/0000-0002-6143-1520 NR 78 TC 34 Z9 35 U1 1 U2 2 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD JUN PY 2002 VL 301 IS 3 BP 1025 EP 1032 AR UNSP 33704/986124 DI 10.1124/jpet.301.3.1025 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LK UT WOS:000175736800031 PM 12023534 ER PT J AU Baek, SJ Wilson, LC Lee, CH Eling, TE AF Baek, SJ Wilson, LC Lee, CH Eling, TE TI Dual function of nonsteroidal anti-inflammatory drugs (NSAIDs): Inhibition of cyclooxygenase and induction of NSAID-activated gene SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID TGF-BETA SUPERFAMILY; CARCINOMA CELL-LINES; FAMILIAL ADENOMATOUS POLYPOSIS; ANTIINFLAMMATORY DRUGS; CANCER CELLS; MORPHOGENETIC PROTEIN; PROSTATE-CANCER; COLON-CANCER; APOPTOSIS; GROWTH AB Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used drugs for the treatment of inflammatory disease and have a chemopreventive effect on colorectal cancer. NSAIDs inhibit cyclooxygenase (COX)-1 and/or COX-2 activity, but the chemopreventive effect may be, in part, independent of prostaglandin inhibition. NSAID-activated gene (NAG-1) was previously identified as a gene induced by some NSAIDs in cells devoid of COX activity. NAG-1 has proapoptotic and antitumorigenic activity in vitro and in vivo. To determine whether the induction of NAG-1 by NSAIDs is influenced by COX expression, we developed COX-1- and COX-2-overexpressing HCT-116 cells. COX expression did not affect NSAID-induced NAG-1 expression as assessed by transient and stable transfection. Also, NAG-1 expression was not affected by PGE(2) and arachidonic acid, suggesting that NAG-1 induction by NSAIDs occurs by a prostanoid-independent manner. We also report that indomethacin increased NAG-1 expression in a number of cells from tissues other than colorectal. In conclusion, NSAIDs have dual function, induction of NAG-1 expression and inhibition of COX activity that occurs in a variety of cell lines. C1 NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Eling, TE (reprint author), NIEHS, Mol Carcinogenesis Lab, NIH, 111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. OI Baek, Seung/0000-0001-7866-7778 NR 45 TC 93 Z9 95 U1 0 U2 1 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD JUN PY 2002 VL 301 IS 3 BP 1126 EP 1131 AR UNSP 33068/986118 DI 10.1124/jpet.301.3.1126 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LK UT WOS:000175736800043 PM 12023546 ER PT J AU Baumann, MH Ayestas, MA Sharpe, LG Lewis, DB Rice, KC Rothman, RB AF Baumann, MH Ayestas, MA Sharpe, LG Lewis, DB Rice, KC Rothman, RB TI Persistent antagonism of methamphetamine-induced dopamine release in rats pretreated with GBR12909 decanoate SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID MONOAMINE REUPTAKE INHIBITORS; MEDICATION DEVELOPMENT; I-125 RTI-55; COCAINE; AMPHETAMINE; GBR-12909; SEROTONIN; FENFLURAMINE; TRANSPORTERS; PHENTERMINE AB Methamphetamine abuse is a serious global health problem, and no effective treatments for methamphetamine dependence have been developed. In animals, the addictive properties of methamphetamine are mediated via release of dopamine (DA) from nerve terminals in mesolimbic reward circuits. At the molecular level, methamphetamine promotes DA release by a nonexocytotic diffusion-exchange process involving DA transporter (DAT) proteins. We have shown that blocking DAT activity with high-affinity DA uptake inhibitors, such as 1-[2[bis( 4-fluorophenyl) methoxy] ethyl]-4-(3-phenylpropyl) piperazine (GBR12909), can substantially reduce amphetamine-induced DA release in vivo. In the present study, we examined the ability of a long-acting depot formulation of GBR12909 decanoate (GBR-decanoate) to influence neurochemical actions of methamphetamine in the nucleus accumbens of rats. Rats received single injections of GBR-decanoate (480 mg/kg i.m.) and were subjected to in vivo microdialysis testing 1 and 2 weeks later. Pretreatment with GBR-decanoate produced modest elevations in basal extracellular levels of DA, but not 5-hydroxytryptamine (5-HT), at both time points. GBR-decanoate nearly eliminated the DA-releasing ability of methamphetamine (0.3 and 1.0 mg/kg i. v.) for 2 weeks, whereas methamphetamine-induced 5-HT release was unaffected. Autoradiographic analysis revealed that GBR-decanoate caused long-term decreases in DAT binding in the brain. Our data suggest that GBR-decanoate, or similar agents, may be useful adjuncts in treating methamphetamine dependence. This therapeutic strategy would be especially useful for noncompliant patient populations. C1 NIDA, Clin Psychopharmacol Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. NIDA, Behav Neurosci Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Baumann, MH (reprint author), NIDA, Clin Psychopharmacol Sect, Intramural Res Program, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 44 TC 26 Z9 26 U1 0 U2 1 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD JUN PY 2002 VL 301 IS 3 BP 1190 EP 1197 AR UNSP 4742/987466 DI 10.1124/jpet.301.3.1190 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LK UT WOS:000175736800051 PM 12023554 ER PT J AU Webster, EL Barrientos, RM Contoreggi, C Isaac, MG Ligier, S Gabry, KE Chrousos, GP McCarthy, EF Rice, KC Gold, PW Sternberg, EM AF Webster, EL Barrientos, RM Contoreggi, C Isaac, MG Ligier, S Gabry, KE Chrousos, GP McCarthy, EF Rice, KC Gold, PW Sternberg, EM TI Corticotropin releasing hormone (CRH) antagonist attenuates adjuvant induced arthritis: Role of CRH in peripheral inflammation SO JOURNAL OF RHEUMATOLOGY LA English DT Article DE corticotropin releasing receptor antagonist; rats; inflammation; adjuvant induced arthritis; glucocorticoids ID PITUITARY-ADRENAL AXIS; MAST-CELL DEGRANULATION; LEWIS RATS; RHEUMATOID-ARTHRITIS; RECEPTOR ANTAGONIST; STRESS-RESPONSE; NEUROENDOCRINE; SUSCEPTIBILITY; DISEASE; SYSTEM AB Objective. To determine whether a corticotropin releasing hormone (CRH) type 1-specific receptor antagonist, antalarmin, would alter the progression of inflammation in adjuvant induced arthritis (AIA) susceptible LEW/N rats by blocking local CRH mediated inflammatory responses or render AIA resistant F344/N rats more susceptible to AIA by blocking central CRH, thus reducing secretion of endogenous glucocorticoids. Methods. F344/N and LEW/N rats were assigned to either drug or vehicle groups and treated with 20 mg/kg antalarmin or vehicle alone BID for 25 days by intraperitoneal injection. Arthritis was induced in both antalarmin and vehicle treated LEW/N and F344/N rats by subcutaneous injections at the base of the tail of incomplete Freund's adjuvant containing 10 mg/ml heat killed Mycobacterium tuberculosis. Control F344/N and LEW/N rats were maintained on either antalarmin or vehicle. Results. Chronic blockade of CRH-R1 with systemic antalarmin significantly ameliorated AIA in LEW/N rats, reducing the severity of inflammation in peripheral joints, evidenced by clinical and histopathology scores, and weight loss associated with disease onset. Antalarmin neither induced nor exacerbated arthritis expression in F344/N or LEW/N rats, despite suppression of levels of adjuvant induced corticosterone, the major antiinflammatory glucocorticoid in rats. Conclusion. Systemic blockade of CRH-R1 appeared to predominantly block peripheral proinflammatory effects of immune CRH, rather than the systemic glucocorticoid mediated anti inflammatory effects of hypothalamic CRH. Results indicate that chronic treatment with a CRH antagonist attenuates progressive inflammation induced degeneration of synovia, cartilage, and bone in arthritic joints, suggesting that antalarmin may have therapeutic potential in treatment of human autoimmune and inflammatory disorders. C1 NIMH, Integrat Neural Immune Program, Sect Neuroendocrine Immunol & Behav, NIH, Bethesda, MD 20892 USA. NIMH, Clin Neuroendocrinol Branch, NIH, Bethesda, MD 20892 USA. NIDA, Brain Imaging Unit, Baltimore, MD USA. NICHHD, Pediat Endocrinol Sect, PREB, Bethesda, MD USA. Johns Hopkins Univ, Sch Med, Dept Orthoped Surg, Baltimore, MD USA. NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Sternberg, EM (reprint author), NIMH, Integrat Neural Immune Program, Sect Neuroendocrine Immunol & Behav, NIH, 36 Convent Dr,MSC 4020,Bldg 36, Bethesda, MD 20892 USA. RI Barrientos, Ruth/E-3244-2013 OI Barrientos, Ruth/0000-0001-7224-4109 NR 38 TC 53 Z9 54 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 JUN PY 2002 VL 29 IS 6 BP 1252 EP 1261 PG 10 WC Rheumatology SC Rheumatology GA 559PA UT WOS:000176033500024 PM 12064844 ER PT J AU Chi-Fishman, G Sonies, BC AF Chi-Fishman, G Sonies, BC TI Kinematic strategies for hyoid movement in rapid sequential swallowing SO JOURNAL OF SPEECH LANGUAGE AND HEARING RESEARCH LA English DT Article DE sequential swallowing; hyoid bone; kinematics; ultrasound; gender ID UPPER ESOPHAGEAL SPHINCTER; VELOCITY; SPEECH; VOLUME AB Past videofluoroscopic and EMG evidence has shown that rapid sequential Swallowing differs from discrete swallows, but our knowledge of the control strategies remains incomplete. This study examined in detail the interrelationships among kinematic variables to discern the strategies for deglutitive hyoid motion during discrete (5 cc, 10 cc, 20 cc, 30 cc) and rapid sequential (120 cc) swallowing tasks. Submental ultrasound was conducted with head and transducer stabilization on 30 healthy subjects (15 males, 15 females) in three age groups (20-39, 40-59, 60-79 yrs). Frame-by-frame changes in hyoid position were tracked from digitized images of 236 discrete and 318 rapid sequential swallows. Repeated-measures analyses of variance were conducted on a number of kinematic variables with corrections for multiple tests and comparisons. The main effect of task was significant for all variables except forward peak velocity. Per post hoc contrasts, rapid sequential swallows had significantly reduced maximal amplitude (maximal displacement), total distance, backward peak velocity at-max and total durations, and time to backward peak velocity in comparison with discrete swallows of any volume. Amplitude "down-scaling" was the prominent kinematic strategy used to accomplish rapid sequential swallows in a shorter time while keeping forward peak velocity essentially unchanged. In contrast, amplitude "up-scaling" was the strategy for accommodating larger-volume discrete swallows. Our results confirm built-in flexibility in the functional range of deglutitive hyoid motion. C1 NIH, Ctr Clin, Bethesda, MD 20892 USA. RP Chi-Fishman, G (reprint author), NIH, Ctr Clin, Rm 6s235,Bldg 10,OMF-PDB-RMD,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 23 TC 17 Z9 22 U1 0 U2 0 PU AMER SPEECH-LANGUAGE-HEARING ASSOC PI ROCKVILLE PA 10801 ROCKVILLE PIKE, ROCKVILLE, MD 20852-3279 USA SN 1092-4388 J9 J SPEECH LANG HEAR R JI J. Speech Lang. Hear. Res. PD JUN PY 2002 VL 45 IS 3 BP 457 EP 468 DI 10.1044/1092-4388(2002/036) PG 12 WC Audiology & Speech-Language Pathology; Linguistics; Rehabilitation SC Audiology & Speech-Language Pathology; Linguistics; Rehabilitation GA 561MH UT WOS:000176144200004 PM 12068999 ER PT J AU DeRijk, RH Schaaf, M de Kloet, ER AF DeRijk, RH Schaaf, M de Kloet, ER TI Glucocorticoid receptor variants: clinical implications SO JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY LA English DT Review DE glucocorticoid receptor variants; clinical implications; hypothalamic-pituitary-adrenal axis ID PRIMARY CORTISOL RESISTANCE; FRAGMENT-LENGTH-POLYMORPHISM; HORMONE-BINDING DOMAIN; HUMAN MULTIPLE-MYELOMA; BETA-ISOFORM; DNA-BINDING; MESSENGER-RNA; TRANSCRIPTIONAL ACTIVATION; RESPONSE ELEMENT; CUSHINGS-DISEASE AB Following exposure to stress, cortisol is secreted from the adrenal cortex under the control of the hypothalamic-pituitary-adrenal axis (HPA-axis). Central in the regulation of the HPA-axis is a two tied corticosteroid-receptor system, comprised of high and low affinity receptors, the mineralocorticoid receptor (MR) and the glucocorticoid receptor (GR), respectively. In addition, these corticosteroid receptors mediate the effects of cortisol during stress on both central and peripheral targets. Cortisol modulates gene-expression of corticosteroid-responsive genes, with the effect lasting from hours to days. Mutations in the GR-gene are being associated with corticosteroid resistance and haematological malignancies, although these mutations are relatively rare and probably not a common cause of these diseases. However, several GR-gene variants and single nucleotide polymorphisms (SNP) in the GR-gene have been identified which are relatively common in the human population. The GRbeta-variant, for example, has been proposed to influence corticosteroid-sensitivity and most evidence has been derived from the immune system and in particular asthma. With respect to polymorphisms, a BclI restriction fragment polymorphism and a Asp363Ser have been described, which not only influence the regulation of the HPA-axis, but are also associated with changes in metabolism and cardiovascular control. These associations of a GR-gene polymorphism with metabolism and cardivascular control, and also with the regulation of the HPA-axis, indicates an important underlying role of cortisol in the etiology of these complex disorders. Therefore, we propose that a common underlying defect in these complex disorders is a disregulation of the HPA-axis, especially during stress. The clinical implication is that the regulation of the HPA-axis should be envisioned as a primary target of new drugs for the treatment of stress-related disorders. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Rijngeestgroep LUMC, Hosp Psychiat, Dept Psychiat, NL-2342 AJ Oesgstsgeest, Netherlands. NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC USA. LUMC, Dept Med Pharmacol, Oesgstsgeest, Netherlands. RP DeRijk, RH (reprint author), Rijngeestgroep LUMC, Hosp Psychiat, Dept Psychiat, Endegeesterstr Weg 5, NL-2342 AJ Oesgstsgeest, Netherlands. NR 151 TC 163 Z9 169 U1 1 U2 7 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-0760 J9 J STEROID BIOCHEM JI J. Steroid Biochem. Mol. Biol. PD JUN PY 2002 VL 81 IS 2 BP 103 EP 122 AR PII S0960-0760(02)00062-6 DI 10.1016/S0960-0760(02)00062-6 PG 20 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 592MT UT WOS:000177942000001 PM 12137800 ER PT J AU Zimmerman, SB AF Zimmerman, SB TI Toroidal nucleoids in Escherichia coli exposed to chloramphenicol SO JOURNAL OF STRUCTURAL BIOLOGY LA English DT Article DE chloramphenicol; Escherichia coli; nucleoid; toroid ID PROTEIN-SYNTHESIS; LIGHT-MICROSCOPY; CELL-DIVISION; DNA; CONDENSATION; ORGANIZATION; INHIBITION; FILAMENTS; BACTERIA; RELEASE AB The DNA of growing cells of Escherichia coli occurs in one or a few lobular bodies known as nucleoids. Upon exposure to chloramphenicol, the nucleoids assume compact, rounded forms ("cm-nucleoids") that have been described as ring- or sphere-shaped. Multiple views of single cells or spheroplasts, however, support a different, curved toroid shape for cm-nucleoids. The multiple views were obtained either by DNA fluorescence imaging as the cells or spheroplasts reoriented in liquid medium or by optical sectioning using phase-contrast or fluorescence imaging of immobilized cells. The curved toroid shape is consistent with electron microscope images of thin sections of chloramphenicol-treated cells. The relationship of this structure to active and inactive nucleoids and to the smaller toroidal forms made by in vitro DNA condensation is discussed. Published by Elsevier Science (USA). C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Zimmerman, SB (reprint author), NIDDKD, Mol Biol Lab, NIH, Bldg 5,Room 329, Bethesda, MD 20892 USA. NR 38 TC 15 Z9 15 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1047-8477 J9 J STRUCT BIOL JI J. Struct. Biol. PD JUN PY 2002 VL 138 IS 3 BP 199 EP 206 AR PII S1047-8477(02)00036-9 DI 10.1016/S1047-8477(02)00036-9 PG 8 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 596BT UT WOS:000178144400004 PM 12217658 ER PT J AU Owens, PL Hoagwood, K Horwitz, SM Leaf, PJ Poduska, JM Kellam, SG Ialongo, NS AF Owens, PL Hoagwood, K Horwitz, SM Leaf, PJ Poduska, JM Kellam, SG Ialongo, NS TI Barriers to children's mental health services SO JOURNAL OF THE AMERICAN ACADEMY OF CHILD AND ADOLESCENT PSYCHIATRY LA English DT Article DE barriers to care; mental health ID DIAGNOSTIC INTERVIEW SCHEDULE; PSYCHIATRIC-DISORDERS; ADOLESCENTS; RELIABILITY; BEHAVIOR; YOUTH; CARE AB Objective: To examine the characteristics associated with barriers to children's mental health services, focusing on the effect of children's psychosocial problems on parents. Method: Data come from a first-grade, prevention-intervention project conducted in Baltimore, Maryland. Analyses were restricted to 116 families who participated in seventh-grade interviews and indicated the index child needed services. The Services Assessment for Children and Adolescents was used to measure barriers to children's mental health services. Results: More than 35% of parents reported a barrier to mental health services. Types of barriers included those related to structural constraints, perceptions of mental health, and perceptions of services (20.7%, 23.3%, and 25.9%, respectively). Although parenting difficulties were associated with all barriers (structural: OR = 10.63, 95% CI: 2.37, 47.64; mental health: OR = 8.31, 95% CI: 1.99, 34.79; services: OR = 5.22, 95% CI: 1.56, 17.51), additional responsibilities related to attendance at meetings was associated only with structural barriers (OR = 5,49, 95% CI: 1,22, 24.59). Conclusions: Researchers and policymakers interested in increasing children's access to mental health services should consider strategies to reduce barriers related to perceptions about mental health problems and services, in addition to structural barriers. Particular attention should be given to programs that focus on the needs of families who are most affected by their child's psychosocial problems. C1 Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Mental Hyg, Baltimore, MD USA. NIMH, Rockville, MD 20857 USA. Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, New Haven, CT 06510 USA. Amer Inst Res, Washington, DC USA. RP Owens, PL (reprint author), Agcy Healthcare Res & Qual, Ctr Org & Delivery Studies, 2101 E Jefferson St,Suite 605, Rockville, MD 20852 USA. FU NIMH NIH HHS [5P0-MH38725, 5T32-MH19545, 2P50-MH43703] NR 30 TC 168 Z9 169 U1 4 U2 13 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 JUN PY 2002 VL 41 IS 6 AR UNSP 0890-8567/02/4106-0731 DI 10.1097/00004583-200206000-00013 PG 8 WC Psychology, Developmental; Pediatrics; Psychiatry SC Psychology; Pediatrics; Psychiatry GA 554PM UT WOS:000175744700013 PM 12049448 ER PT J AU Hirshberg, B Libutti, SK Alexander, HR Bartlett, BL Cochran, C Livi, A Chang, R Shawker, T Skarulis, MC Gorden, P AF Hirshberg, B Libutti, SK Alexander, HR Bartlett, BL Cochran, C Livi, A Chang, R Shawker, T Skarulis, MC Gorden, P TI Blind distal pancreatectomy for occult insulinoma, an inadvisable procedure SO JOURNAL OF THE AMERICAN COLLEGE OF SURGEONS LA English DT Article; Proceedings Paper CT 86th Annual Clinical Congress of the American-College-of-Surgeons CY OCT 23-24, 2000 CL CHICAGO, ILLINOIS SP Amer Coll Surgeons ID INTRAOPERATIVE ULTRASONOGRAPHY; HYPOGLYCEMIC DISORDERS; LOCALIZATION AB BACKGROUND: Fasting hypoglycemia with neuroglycopenic symptoms corrected by administration of glucose are the hallmarks for the diagnosis of insulinoma. Surgical resection is the treatment of choice for insulinomas, but localization of these lesions can be challenging. Blind distal pancreatectomy has been advocated for occult insulinomas not detected on imaging studies or during abdominal exploration. Wide the advent of newer localization techniques, we challenge the wisdom of this approach. STUDY DESIGN: The records of patients (multiple endocrine neoplasia excluded) with pathologically proved insulinoma who were screened at our institution or referred to us after a failed blind distal pancreatectomy were reviewed. All records included patient history and results of physical examination and routine blood and urine tests. The diagnosis of insulinoma was confirmed during a supervised fast. Patients with biochemically proved insulinoma underwent localization studies and operation. Studies included CT scans, MRI, transabdominal ultrasound, intraoperative ultrasonography, angiography (more recently, Ca++-stimulated arteriography), and venous sampling. RESULTS: From 1970 to 2000, 99 patients (34 men, 65 women; mean age 43 years) underwent operation. All patients with benign tumors (92) were cured after operation. Seventeen patients were referred to the NIH after a failed blind distal pancreatectomy. Of these, 5 were diagnosed as having factitious hypoglycemia. In the other 12 patients a tumor was localized in the pancreatic head. Two patients incorrectly diagnosed with nesidioblastosis after initial surgery were subsequently cured by resection of an insulinoma. CONCLUSIONS: The use of preoperative imaging studies, most notably Ca++-stimulated arteriography, and intraoperative ultrasonography permits detection of virtually all insulinomas, including reoperated cases. When a tumor is not detected, the procedure should be terminated and the patient referred to a center capable of performing advanced preoperative and intraoperative localization techniques. With the preoperative and intraoperative imaging strategies currently available, the use of blind distal pancreatectomy for occult insulinoma should be abolished. C1 NCI, Surg Metab Sect, Surg Branch, NIH,Ctr Canc Res, Bethesda, MD 20892 USA. NIDDK, Div Intramural Res, NIH, Bethesda, MD USA. NIH, Dept Radiol, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Libutti, SK (reprint author), NCI, Surg Metab Sect, Surg Branch, NIH,Ctr Canc Res, 10 Ctr Dr,MSC 1502,Bldg 10,Rm 2B07, Bethesda, MD 20892 USA. NR 24 TC 54 Z9 63 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1072-7515 J9 J AM COLL SURGEONS JI J. Am. Coll. Surg. PD JUN PY 2002 VL 194 IS 6 BP 761 EP 764 AR PII S1072-7515(02)01177-8 DI 10.1016/S1072-7515(02)01177-8 PG 4 WC Surgery SC Surgery GA 562NM UT WOS:000176204800014 PM 12081066 ER PT J AU Stables, GJ Subar, AF Patterson, BH Dodd, K Heimendinger, J Van Duyn, MAS Nebeling, L AF Stables, GJ Subar, AF Patterson, BH Dodd, K Heimendinger, J Van Duyn, MAS Nebeling, L TI Changes in vegetable and fruit consumption and awareness among US adults: Results of the 1991 and 1997 5 A Day for Better Health Program surveys SO JOURNAL OF THE AMERICAN DIETETIC ASSOCIATION LA English DT Article ID INCREASING FRUIT; CANCER PREVENTION; TREATWELL 5-A-DAY; KNOWLEDGE; DIET; RISK; QUESTIONNAIRE; WORKSITE; CHILDREN; PROJECT AB Objective The purpose of this study is to assess population-based changes in vegetable and fruit consumption and psychosocial correlates. Design Two nationally representative random digit dial surveys conducted in 1991 and 1997; respondents were queried regarding consumption of and attitudes and knowledge about vegetables and fruit. Subjects/Setting Respondents were 2,755 and 2,544 adults (in 1991 and 1997, respectively) older than 18 years. Statistical Analysis Performed Vegetable and fruit consumption and message awareness were measured using weighted-only and regression model-adjusted analyses to assess changes. Results Mean vegetable and fruit consumption was significantly (P=.007) higher in 1997 than in 1991 using weighted-only analyses, but remained significant only for Hispanic (P=.03) and nonsmoker (P=.004) subgroups when adjusted for demographic shifts. Significantly higher percentages were found in the model-adjusted analyses for those consuming 5 or more daily servings (23.4% to 25.8%), message awareness (7.7% to 19.2%), and knowledge of the 5 A Day Program (2.0% to 17.8%). Applications/Conclusions A significantly positive change in vegetable and fruit consumption occurred between 1991 and 1997 according to traditional methods of survey data analysis, but null findings resulted when the data were adjusted for demographic shifts. Nutrition professionals should continue targeting specific demographic subgroups with tailored interventions to move all Americans toward achievement of dietary guidelines for vegetable and fruit consumption. C1 NCI, Appl Res Program, Risk Factor Monitoring & Methods Branch, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. NCI, Div Canc Control & Populat Sci, Stat Res & Applicat Branch, Bethesda, MD 20892 USA. NCI, Div Canc Control & Populat Sci, Hlth Promot Res Branch, Bethesda, MD 20892 USA. NCI, Div Canc Prevent, Biometry Res Grp, Bethesda, MD 20892 USA. AMC Canc Res Ctr & Fdn, Denver, CO USA. NCI, Office Commun, Hlth Promot Branch, Bethesda, MD 20892 USA. RP Stables, GJ (reprint author), NCI, Div Canc Control & Populat Sci, Hlth Promot Res Branch, Behav Res Program, 6130 Execut Blvd,Room 4052, Bethesda, MD 20892 USA. NR 42 TC 104 Z9 109 U1 1 U2 9 PU AMER DIETETIC ASSOC PI CHICAGO PA 216 W JACKSON BLVD #800, CHICAGO, IL 60606-6995 USA SN 0002-8223 J9 J AM DIET ASSOC JI J. Am. Diet. Assoc. PD JUN PY 2002 VL 102 IS 6 BP 809 EP 817 DI 10.1016/S0002-8223(02)90181-1 PG 9 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 560DZ UT WOS:000176068500015 PM 12067046 ER PT J AU Alroy, J Sabnis, S Kopp, JB AF Alroy, J Sabnis, S Kopp, JB TI Renal pathology in Fabry disease SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article ID ALPHA-GALACTOSIDASE; TRANSPLANTATION; ALLOGRAFT; HISTOCHEMISTRY; RECURRENCE; STORAGE C1 NIDDKD, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. Tufts Univ, Sch Vet Med, Dept Pathol, Boston, MA 02111 USA. Tufts Univ, Sch Med, Dept Pathol, Boston, MA 02111 USA. Tufts Univ New England Med Ctr, Boston, MA 02111 USA. Armed Forces Inst Pathol, Div Nephrol, Washington, DC 20306 USA. RP Kopp, JB (reprint author), NIDDKD, Kidney Dis Sect, NIH, 10-3N116, Bethesda, MD 20892 USA. NR 21 TC 71 Z9 88 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 JUN PY 2002 VL 13 IS 6 SU 2 BP S134 EP S138 AR UNSP 1046-6673/1300-0134 DI 10.1097/01.ASN.0000016684.07368.75 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 558YZ UT WOS:000175997000004 PM 12068025 ER PT J AU Branton, M Schiffmann, R Kopp, JB AF Branton, M Schiffmann, R Kopp, JB TI Natural history and treatment of renal involvement in Fabry disease SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article ID REPLACEMENT THERAPY; ENZYME REPLACEMENT; SAFETY; TRIAL C1 NIDDKD, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. NINCDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Kopp, JB (reprint author), NIDDKD, Kidney Dis Sect, NIH, 10-3N116, Bethesda, MD 20892 USA. NR 13 TC 55 Z9 60 U1 0 U2 3 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 JUN PY 2002 VL 13 IS 6 SU 2 BP S139 EP S143 AR UNSP 1046-6673/1300-0139 DI 10.1097/01.ASN.0000016683.73778.78 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 558YZ UT WOS:000175997000005 PM 12068026 ER PT J AU Roberson, R AF Roberson, R TI Leap of faith into the millennium SO JOURNAL OF THE NATIONAL MEDICAL ASSOCIATION LA English DT Letter C1 NIH, Baltimore, MD USA. RP Roberson, R (reprint author), NIH, Baltimore, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATL MED ASSOC PI WASHINGON PA 1012 10TH ST, N W, WASHINGON, DC 20001 USA SN 0027-9684 J9 J NATL MED ASSOC JI J. Natl. Med. Assoc. PD JUN PY 2002 VL 94 IS 6 BP 513 EP 514 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 562DN UT WOS:000176182300024 PM 12078933 ER PT J AU Zhang, QL Liu, J Lin, PX Webster, HD AF Zhang, QL Liu, J Lin, PX Webster, HD TI Local administration of vasoactive intestinal peptide after nerve transection accelerates early myelination and growth of regenerating axons SO JOURNAL OF THE PERIPHERAL NERVOUS SYSTEM LA English DT Article DE vasoactive intestinal peptide; nerve regeneration; laminin; Schwann cell; myelin ID CILIARY NEUROTROPHIC FACTOR; SCHWANN-CELL PROLIFERATION; SCIATIC-NERVE; SYMPATHETIC NEURONS; ELECTRON-MICROSCOPY; LAMININ PRODUCTION; GENE-EXPRESSION; RAT; DEGENERATION; POLYPEPTIDE AB Our goal was to determine whether local injections of vasoactive intestinal peptide (VIP) promote early stages of regeneration after nerve transection. Sciatic nerves were transected bilaterally in 2 groups of 10 adult mice. In the first group, 15 mug (20 muL) of VIP were injected twice daily into the gap between transected ends of the right sciatic nerve for 7 days (4 mice) or 14 days (6 mice). The same number of mice in the second group received placebo injections (20 muL of 0.9% sterile saline) in the same site, twice daily, for the same periods. After 7 days, axon sizes, relationships with Schwann cells and degree of myelination were compared in electron micrographs of transversely sectioned distal ends of proximal stumps. Fourteen days after transection, light and electron microscopy were used to compare and measure axons and myelin sheaths in the transection gap, 2-mm distal to the ends of proximal stumps. Distal ends of VIP-treated proximal stumps contained larger axons 7 days after transection. More axons were in 1:1 relationships with Schwann cells and some of them were surrounded by thin myelin sheaths. In placebo-treated proximal slumps, axons were smaller, few were in 1:1 relationships with Schwann cells and no myelin sheaths were observed. In VIP-treated transection gaps, measurements 14 days after transection showed that larger axons were more numerous and their myelin sheaths were thicker. Our results suggest that in this nerve transection model, local administration of VIP promotes and accelerates early myelination and growth of regenerating axons. C1 NINCDS, Basic Neurosci Program, NIH, Bethesda, MD 20892 USA. NINCDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Webster, HD (reprint author), NINCDS, Basic Neurosci Program, NIH, Bldg 36,Rm 4A-29, Bethesda, MD 20892 USA. NR 51 TC 11 Z9 12 U1 0 U2 1 PU WOODLAND PUBLICATIONS, INC PI NEW YORK PA 69 MURRAY ST, NEW YORK, NY 10007 USA SN 1085-9489 J9 J PERIPHER NERV SYST JI J. Peripher. Nerv. Syst. PD JUN PY 2002 VL 7 IS 2 BP 118 EP 127 DI 10.1046/j.1529-8027.2002.02018.x PG 10 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 563ZX UT WOS:000176289600005 PM 12090298 ER PT J AU Lee, W Chaiworapongsa, T Romero, R Williams, R McNie, B Johnson, A Treadwell, M Comstock, CH AF Lee, W Chaiworapongsa, T Romero, R Williams, R McNie, B Johnson, A Treadwell, M Comstock, CH TI A diagnostic approach for the evaluation of spina bifida by three-dimensional ultrasonography SO JOURNAL OF ULTRASOUND IN MEDICINE LA English DT Article DE fetus; neural tube defect; prenatal diagnosis; spina bifida; three-dimensional ultrasonography ID NEURAL-TUBE DEFECTS; 3-DIMENSIONAL ULTRASOUND; FETAL SPINE; PRENATAL ULTRASOUND; FETUSES; MALFORMATIONS; OSSIFICATION; ANOMALIES; LEVEL AB Objective. To describe a prenatal diagnostic method for evaluating spina bifida by three-dimensional ultrasonography, Methods. Two- and three-dimensional ultrasonography were used to determine the extent of vertebral defects among fetuses with spina bifida. Spinal levels were independently counted from the most caudal thoracic vertebra with a rib (e.g., 12th thoracic rib), A virtual cutting plane was manipulated through a volume-rendered spine to generate optimal multiplanar views for this blinded analysis, Prenatal diagnosis was compared with a postnatal analysis of bony spine defects derived from radiographic films or magnetic resonance imaging. Results. Nine fetuses were examined by two-dimensional ultrasonography (21.8 +/- 3.4 menstrual weeks) and three-dimensional ultrasonography (22,8 +/- 4.4 menstrual weeks), For two-dimensional ultrasonography, the spinal level agreed to within 1 vertebral segment in 6 of 9 infants, in contrast, three-dimensional ultrasonography agreed to within 1 vertebral segment in 8 of 9 infants. Three fetuses had vertebral defect levels on two-dimensional ultrasonography that were 1,5 to 2 segments away from postnatal findings, The same fetuses had results that were within 1 vertebral segment on three-dimensional ultrasonography, Volume rendering showed splayed vertebral pedicles and disrupted vertebrae. An intact meningeal sac was easily rendered in 5 of 9 subjects, Conclusions. Multiplanar views are generally more informative than rendered views for localizing bony defects of the fetal spine. The level of the defect on three-dimensional ultrasonography correlates well with those on two-dimensional ultrasonography and postnatal imaging studies. This approach may improve characterization of spina bifida by adding diagnostic information that is complementary to the initial assessment by two-dimensional ultrasonography. C1 William Beaumont Hosp, Dept Obstet & Gynecol, Div Fetal Imaging, Royal Oak, MI 48073 USA. NICHHD, Perinatol Res Branch, Detroit, MI USA. Wayne State Univ, Hutzel Hosp, Dept Obstet & Gynecol, Detroit, MI USA. RP Lee, W (reprint author), William Beaumont Hosp, Dept Obstet & Gynecol, Div Fetal Imaging, 3601 W 13 Mile Rd, Royal Oak, MI 48073 USA. NR 22 TC 23 Z9 26 U1 1 U2 1 PU AMER INST ULTRASOUND MEDICINE PI LAUREL PA SUBSCRIPTION DEPT, 14750 SWEITZER LANE, STE 100, LAUREL, MD 20707-5906 USA SN 0278-4297 J9 J ULTRAS MED JI J. Ultrasound Med. PD JUN PY 2002 VL 21 IS 6 BP 619 EP 626 PG 8 WC Acoustics; Radiology, Nuclear Medicine & Medical Imaging SC Acoustics; Radiology, Nuclear Medicine & Medical Imaging GA 556FV UT WOS:000175839500001 PM 12054297 ER PT J AU Wright, EJ Fang, JY Metter, EJ Partin, AW Landis, P Chan, DW Carter, HB AF Wright, EJ Fang, JY Metter, EJ Partin, AW Landis, P Chan, DW Carter, HB TI Prostate specific antigen predicts the long-term risk of prostate enlargement: Results from the Baltimore Longitudinal Study of Aging SO JOURNAL OF UROLOGY LA English DT Article DE prostate-specific antigen; prostatic hyperplasia; prostate ID ACUTE URINARY RETENTION; MEN; HYPERPLASIA; GROWTH AB Purpose: Prostate specific antigen (PSA) is a predictor of prostate growth in men with lower urinary tract symptoms. The long-term risk of prostate enlargement as a function of PSA among community dwelling volunteers is unknown. Materials and Methods: A Cox proportional hazards regression model was used to study the relationship between baseline PSA level at ages 40 to 49.9 years in 194 men, 50 to 59.9 in 191 and 60 to 69.9 in 144, and prostate enlargement, defined as a prostate volume larger than the 75th percentile for age decade, as measured by magnetic resonance imaging in a longitudinal study of aging (Baltimore Longitudinal Study of Aging, National Institute on Aging). Kaplan-Meier survival analysis was performed to estimate the probability of freedom from prostate enlargement with time as a function of baseline PSA level. Results: The relative risk of prostate enlargement was 3- to 6-fold higher for men 40 to 49.9 years old with a baseline PSA of 0.31 ng./ml. or more compared to men with PSA levels of 0.30 ng./ml. or less at baseline. The relative risk was increased 5- to 9-fold in men 50 to 59.9 years old and 11-fold in those 60 to 69.9 years old when comparing men with PSA greater than 0.80 ng./ml. and greater than 1.70 ng./ml. with those with PSA 0.50 or less. The cumulative probability of freedom from prostate enlargement at 20 years was 0.89 (95% confidence interval ([CI] 0.79-0.99) and 0.63 (0.52-0.74) for men 40 to 49.9 years old with PSA levels below and above 0.30 ng./ml., respectively. For men 50 to 59.9 years old the 10-year probability of freedom from prostate enlargement was 0.90 (95% CI 0.84-0.96) and 0.59 (0.43-0.74) when PSA levels were below and above 0.80 ng./ml., respectively. At age 60 to 69.9 years the 10-year probability of freedom from prostate enlargement was 0.83 (95% CI 0.72-0.93) and 0.27 (0.09 to 0.48) when PSA levels were below and above 1.70 ng./ml., respectively. Conclusions: These data demonstrate the long-term risk of prostate enlargement by PSA level. Risk stratification based on PSA level may be useful to identify men at greatest risk for adverse events due to prostate enlargement and selection of men for future benign prostatic hyperplasia studies. C1 Johns Hopkins Univ, Sch Med, Dept Urol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. James Buchanan Brady Urol Inst, Baltimore, MD USA. Johns Hopkins Univ Hosp, Baltimore, MD 21287 USA. NIA, NIH, Baltimore, MD 21224 USA. Gerontol Res Ctr, Baltimore, MD USA. RP Wright, EJ (reprint author), Johns Hopkins Univ, Sch Med, Dept Urol, Baltimore, MD 21205 USA. NR 11 TC 32 Z9 36 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-5347 J9 J UROLOGY JI J. Urol. PD JUN PY 2002 VL 167 IS 6 BP 2484 EP 2487 AR UNSP 0022-5347/02/1676-2484/0 DI 10.1016/S0022-5347(05)65010-0 PG 4 WC Urology & Nephrology SC Urology & Nephrology GA 552DD UT WOS:000175602500032 PM 11992063 ER PT J AU Chertova, E Bess, JW Crise, BJ Sowder, RC Schaden, TM Hilburn, JM Hoxie, JA Benveniste, RE Lifson, JD Henderson, LE Arthur, LO AF Chertova, E Bess, JW Crise, BJ Sowder, RC Schaden, TM Hilburn, JM Hoxie, JA Benveniste, RE Lifson, JD Henderson, LE Arthur, LO TI Envelope glycoprotein incorporation, not shedding of surface envelope glycoprotein (gp120/SU), is the primary determinant of SU content of purified human immunodeficiency virus type 1 and simian immunodeficiency virus SO JOURNAL OF VIROLOGY LA English DT Article ID AMINO-ACID SUBSTITUTIONS; SOLUBLE CD4; HIV-1 VIRIONS; NEUTRALIZATION RESISTANCE; CYTOPLASMIC DOMAINS; INFECTED-CELLS; CYCLOPHILIN-A; FULL-LENGTH; V3 LOOP; PROTEINS AB Human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) particles typically contain small amounts of the surface envelope protein (SU), and this is widely believed to be due to shedding of SU from mature virions. We purified proteins from HIV-1 and SIV isolates using procedures which allow quantitative measurements of viral protein content and determination of the ratios of gag- and env-encoded proteins in virions. All of the HIV-1 and most of the SIN isolates examined contained low levels of envelope proteins, with Gag:Env ratios of approximately 60:1. Based on an estimate of 1,200 to 2,500 Gag molecules per virion, this corresponds to an average of between 21 and 42 SU molecules, or between 7 and 14 trimers, per particle. In contrast, some SIV isolates contained levels of SU at least 10-fold greater than SU from HIV-1 isolates. Quantification of relative amounts of SU and transmembrane envelope protein (TM) provides a means to assess the impact of SU shedding on virion SU content, since such shedding would be expected to result in a molar excess of TM over SU on virions that had shed SU. With one exception, viruses with sufficient SU and TM to allow quantification were found to have approximately equivalent molar amounts of SU and TM. The quantity of SU associated with virions and the SU:TM ratios were not significantly changed during multiple freeze-thaw cycles or purification through sucrose gradients. Exposure of purified HIV-1 and SIV to temperatures of 55degreesC or greater for 1 h resulted in loss of most of the SU from the virus but retention of TM. Incubation of purified virus with soluble CD4 at 37degreesC resulted in no appreciable loss of SU from either SIV or HIV-1. These results indicate that the association of SU and TM on the purified virions studied is quite stable. These findings suggest that incorporation of SU-TM complexes into the viral membrane may be the primary factor determining the quantity of SU associated with SIV and HIV-1 virions, rather than shedding of SU from mature virions. C1 NCI, Frederick, MD 21702 USA. SAIC Frederick, AIDS Vaccine Program, Frederick, MD 21702 USA. Univ Penn, Philadelphia, PA 19104 USA. RP Chertova, E (reprint author), NCI, Frederick, MD 21702 USA. RI Bess, Jr., Julian/B-5343-2012 FU NCI NIH HHS [N01-CO-12400, N01CO12400] NR 57 TC 196 Z9 198 U1 0 U2 2 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 JUN PY 2002 VL 76 IS 11 BP 5315 EP 5325 DI 10.1128/JVI.76.11.5315-5325.2002 PG 11 WC Virology SC Virology GA 551EF UT WOS:000175546000003 PM 11991960 ER PT J AU Chakrabarti, BK Kong, W Wu, B Yang, ZY Friborg, J Ling, X King, SR Montefiori, DC Nabel, GJ AF Chakrabarti, BK Kong, W Wu, B Yang, ZY Friborg, J Ling, X King, SR Montefiori, DC Nabel, GJ TI Modifications of the human immunodeficiency virus envelope glycoprotein enhance immunogenicity for genetic immunization SO JOURNAL OF VIROLOGY LA English DT Article ID HIV VACCINES; NEUTRALIZING ANTIBODY; INFECTED MACAQUES; IMMUNE-RESPONSES; TYPE-1; GP41; GP120; EXPRESSION; FUSION; CELLS AB In this study, we have investigated the effect of specific mutations in human immunodeficiency virus type 1 (HIV-1) envelope (Env) on antibody production in an effort to improve humoral immune responses to this glycoprotein by DNA vaccination. Mice were injected with plasmid expression vectors encoding HIV Env with modifications in regions that might affect this response. Elimination of conserved glycosylation sites did not substantially enhance humoral or cytotoxic-T-lymphocyte (CTL) immunity. In contrast, a modified gp140 with different COOH-terminal mutations intended to mimic a fusion intermediate and stabilize trimer formation enhanced humoral immunity without reducing the efficacy of the CTL response. This mutant, with deletions in the cleavage site, fusogenic domain, and spacing of heptad repeats 1 and 2, retained native antigenic conformational determinants as defined by binding to known monoclonal antibodies or CD4, oligomer formation, and virus neutralization in vitro. Importantly, this modified Env, gp140DeltaCFI, stimulated the antibody response to native gp160 while it retained its ability to induce a CTL response, a desirable feature for an AIDS vaccine. C1 NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. Univ Michigan, Dept Internal Med, Ann Arbor, MI 48109 USA. Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA. RP Nabel, GJ (reprint author), NIAID, Vaccine Res Ctr, NIH, 40 Convent Dr, Bethesda, MD 20892 USA. NR 57 TC 124 Z9 126 U1 0 U2 2 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 JUN PY 2002 VL 76 IS 11 BP 5357 EP 5368 DI 10.1128/JVI.76.11.5357-5368.2002 PG 12 WC Virology SC Virology GA 551EF UT WOS:000175546000007 PM 11991964 ER PT J AU Zhang, LQ Peeples, ME Boucher, RC Collins, PL Pickles, RJ AF Zhang, LQ Peeples, ME Boucher, RC Collins, PL Pickles, RJ TI Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology SO JOURNAL OF VIROLOGY LA English DT Article ID GENE-TRANSFER; CYSTIC-FIBROSIS; APICAL SURFACE; ADENOVIRUS INFECTION; PERICILIARY LIQUID; COTTON RATS; GLYCOPROTEIN; ATTACHMENT; RSV; REPLICATION AB Gene therapy for cystic fibrosis (CF) lung disease requires efficient gene transfer to airway epithelial cells after intralumenal delivery. Most gene transfer vectors so far tested have not provided the efficiency required. Although human respiratory syncytial virus (RSV), a common respiratory virus, is known to infect the respiratory epithelium, the mechanism of infection and the epithelial cell type targeted by RSV have not been determined. We have utilized human primary airway epithelial cell cultures that generate a well-differentiated pseudostratified mucociliary epithelium to investigate whether RSV infects airway epithelium via the lumenal (apical) surface. A recombinant RSV expressing green fluorescent protein (rgRSV) infected epithelial cell cultures with high gene transfer efficiency when applied to the apical surface but not after basolateral inoculation. Analyses of the cell types infected by RSV revealed that lumenal columnar cells, specifically ciliated epithelial cells, were targeted by RSV and that cultures became susceptible to infection as they differentiated into a ciliated phenotype. In addition to infection of ciliated cells via the apical membrane, RSV was shed exclusively from the apical surface and spread to neighboring ciliated cells by the motion of the cilial beat. Gross histological examination of cultures infected with RSV revealed no evidence of obvious cytopathology, suggesting that RSV infection in the absence of an immune response can be tolerated for >3 months. Therefore, rgRSV efficiently transduced the airway epithelium via the lumenal surface and specifically targeted ciliated airway epithelial cells. Since rgRSV appears to breach the lumenal barriers encountered by other gene transfer vectors in the airway, this virus may be a good candidate for the development of a gene transfer vector for CF lung disease. C1 Univ N Carolina, Cyst Fibrosis Pulm Res Treatment Ctr, Chapel Hill, NC 27599 USA. Rush Presbyterian St Lukes Med Ctr, Dept Immunol Microbiol, Chicago, IL 60612 USA. NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Zhang, LQ (reprint author), Univ N Carolina, Cyst Fibrosis Pulm Res Treatment Ctr, 7129 Thurston Bowles,CB 7248, Chapel Hill, NC 27599 USA. RI Zhang, Liqun/A-6861-2011 FU NHLBI NIH HHS [HL 51818, P01 HL051818] NR 45 TC 238 Z9 242 U1 1 U2 11 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 JUN PY 2002 VL 76 IS 11 BP 5654 EP 5666 DI 10.1128/JVI.76.11.5654.5666.2002 PG 13 WC Virology SC Virology GA 551EF UT WOS:000175546000037 PM 11991994 ER PT J AU Prikhod'ko, GG Prikhod'ko, EA Pletnev, AG Cohen, JI AF Prikhod'ko, GG Prikhod'ko, EA Pletnev, AG Cohen, JI TI Langat flavivirus protease NS3 binds caspase-8 and induces apoptosis SO JOURNAL OF VIROLOGY LA English DT Article ID DENGUE VIRUS TYPE-2; RNA-STIMULATED NTPASE; YELLOW-FEVER VIRUS; NONSTRUCTURAL PROTEIN; CELL-DEATH; PUTATIVE HELICASES; VIRAL POLYPROTEIN; DOMAIN; CONTAINS; NS2B AB The flavivirus NS3 protein plays an important role in the cleavage and processing of the viral polyprotein and in the synthesis of the viral RNA. NS3 recruits NS2B and NS5 proteins to form complexes possessing protease and replicase activities through protease and nucleoside triphosphatase/helicase domains. We have found that NS3 also induces apoptosis. Expression of the Langat (LGT) virus NS3 protein resulted in a cleavage of cellular DNA and reduced the viability of cells. Coexpression of NS3 with apoptotic inhibitors (CrmA and P35) and addition of caspase peptide substrates (Z-VAD-FMK and Z-IETD-FMK) to NS3-transfected cells blocked NS3-induced apoptosis. In cotransfection experiments, NS3 bound to caspase-8 and enhanced caspase-8-mediated apoptosis. NS3 and caspase-8 colocalized in the cytoplasm of transfected cells. Deletion analysis demonstrated that at least two regions of NS3 contribute to its apoptotic activities. The protease and helicase domains are each able to bind to caspase-8, while the protease domain alone induces apoptosis. The protease domain and tetrahelix region of the helicase domain are required for NS3 to augment caspase-8-mediated apoptosis. Thus, the LGT virus NS3 protein is a multifunctional protein that binds to caspase-8 and induces apoptosis. C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Prikhod'ko, GG (reprint author), Amer Red Cross, Jerome H Holland Lab, Plasma Derivat Dept, 15601 Crabbs Branch Way, Rockville, MD 20855 USA. NR 40 TC 38 Z9 42 U1 1 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 JUN PY 2002 VL 76 IS 11 BP 5701 EP 5710 DI 10.1128/JVI.76.11.5701-5710.2002 PG 10 WC Virology SC Virology GA 551EF UT WOS:000175546000041 PM 11991998 ER PT J AU Wu, L Martin, TD Vazeux, R Unutmaz, D KewalRamani, VN AF Wu, L Martin, TD Vazeux, R Unutmaz, D KewalRamani, VN TI Functional evaluation of DC-SIGN monoclonal antibodies reveals DC-SIGN interactions with ICAM-3 do not promote human immunodeficiency virus type 1 transmission SO JOURNAL OF VIROLOGY LA English DT Article ID C-TYPE LECTIN; DENDRITIC CELLS; T-CELLS; ENDOTHELIAL-CELLS; HIV-1 INFECTION; RHESUS MACAQUE; EXPRESSION; REPLICATION; PROTEIN; TRANS AB DC-SIGN, a type H membrane-spanning C-type lectin that is expressed on the surface of dendritic cells (DC), captures and promotes human and simian immunodeficiency virus (HIV and SIV) infection of CD4(+) T cells in trans. To better understand the mechanism of DC-SIGN-mediated virus transmission, we generated and functionally evaluated a panel of seven monoclonal antibodies (MAbs) against DC-SIGN family molecules. Six of the MAbs reacted with myeloid-lineage DC, whereas one MAb preferentially bound DC-SIGNR/L-SIGN, a homolog of DC-SIGN. Characterization of hematopoietic cells also revealed that stimulation of monocytes with interleukin-4 (IL-4) or IL-13 was sufficient to induce expression of DC-SIGN. All DC-SIGN-reactive MAbs competed with intercellular adhesion molecule 3 (ICAM-3) for adhesion to DC-SIGN and blocked HIV-1 transmission to T cells that was mediated by THP-1 cells expressing DC-SIGN. Similar but less efficient MAb blocking of DC-mediated HIV-1 transmission was observed, indicating that HIV-1 transmission to target cells via DC may not be dependent solely on DC-SIGN. Attempts to neutralize DC-SIGN capture and transmission of HIV-1 with soluble ICAM-3 prophylaxis were limited in success, with a maximal inhibition of 60%. In addition, disrupting DC-SIGN/ICAM-3 interactions between cells with MAbs did not impair DC-SIGN-mediated HIV-1 transmission. Finally, forced expression of ICAM-3 on target cells did not increase their susceptibility to HIV-1 transmission mediated by DC-SIGN. While these findings do not discount the role of intercellular contact in facilitating HIV-1 transmission, our in vitro data indicate that DC-SIGN interactions with ICAM-3 do not promote DC-SIGN-mediated virus transmission. C1 NCI, HIV Drug Resistance Program, NIH, Ft Detrick, MD 21702 USA. ICOS Corp, Bothell, WA 98021 USA. Vanderbilt Univ, Sch Med, Nashville, TN 37232 USA. RP KewalRamani, VN (reprint author), NCI, HIV Drug Resistance Program, NIH, POB B, Ft Detrick, MD 21702 USA. RI Wu, Li/E-4330-2011 OI Wu, Li/0000-0002-5468-2487 FU NIAID NIH HHS [R01 AI042284, R01-AI42284] NR 33 TC 83 Z9 85 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUN PY 2002 VL 76 IS 12 BP 5905 EP 5914 DI 10.1128/JVI.76.12.5905-5914-2002 PG 10 WC Virology SC Virology GA 557MK UT WOS:000175912200007 PM 12021323 ER PT J AU Peng, B Voltan, R Lim, L Edghill-Smith, Y Phogat, S Dimitrov, DS Arora, K Leno, M Than, S Woodward, R Markham, PD Cranage, M Robert-Guroff, M AF Peng, B Voltan, R Lim, L Edghill-Smith, Y Phogat, S Dimitrov, DS Arora, K Leno, M Than, S Woodward, R Markham, PD Cranage, M Robert-Guroff, M TI Rhesus macaque resistance to mucosal simian immunodeficiency virus infection is associated with a postentry block in viral replication SO JOURNAL OF VIROLOGY LA English DT Article ID HIV-1 DISEASE PROGRESSION; CHEMOKINE GENE VARIANT; IN-VITRO; CYCLOPHILIN-A; INTRAVAGINAL INOCULATION; AIDS PROGRESSION; RETROVIRAL DNA; TROPIC HIV-1; TYPE-1 HIV-1; CCR5 AB Elucidation of the host factors which influence susceptibility to human immunodeficiency virus or simian immunodeficiency virus (SIV) infection and disease progression has important theoretical and practical implications. Rhesus macaque 359, a vaccine control animal, resisted two successive intravaginal challenges with SIV(mac251) and failed to seroconvert. Here, after an additional intrarectal SIVmac32H challenge, macaque 359 remained highly resistant to infection. Viral RNA (10(6) copies/ml) was observed in plasma only at week 2 postchallenge. Virus isolation and proviral DNA were positive only once at week eight postchallenge. The animal remained seronegative and cleared SIV in vivo. Its blood and lymph node cells obtained at 49 weeks after intrarectal challenge did not transmit SIV to a naive macaque. We found that the resistance of macaque 359 to SIV infection was not due to a high level of CD8(+) suppressor activity but to an inherent resistance of its CD4(+) T cells. To elucidate the basis for the unusually strong resistance of macaque 359 to SIV infection in vivo and in vitro, we investigated early events of viral infection and replication in CD4(+) cells of macaque 359, including expression and mutation screening of SIV coreceptors and analysis of viral entry and reverse transcription. Mutation screening revealed no genetic alteration in SIV coreceptors. PCR analysis revealed a significant delay in production of early in vitro reverse transcription intermediates in macaque 359 cells compared to susceptible controls, but cell fusion assays showed that SIV entered the CD4(+) CCR5(+) cells of macaque 359 as readily as cells of macaques susceptible to SIV infection. Our results suggest that the resistance of macaque 359 to SIV infection is due to a postentry block in viral replication and implicate a cellular inhibitory mechanism in its CD4(+) T cells. Identification of this host mechanism will help further elucidate the biochemistry of reverse transcription and may suggest therapeutic strategies. Determining the prevalence of this host resistance mechanism among macaques may lead to better design of SIV pathogenesis and vaccine studies. C1 NCI, Basic Res Lab, Bethesda, MD 20892 USA. NCI Frederick, Lab Expt & Computat Biol, NIH, Frederick, MD 21702 USA. Adv BioSci Labs Inc, Kensington, MD 20895 USA. Ctr Appl Microbiol & Res, Salisbury SP4 OJG, Wilts, England. RP Robert-Guroff, M (reprint author), NCI, Basic Res Lab, 41 Lib Dr,Bldg 41,Rm D804, Bethesda, MD 20892 USA. EM guroffm@exchange.nih.gov NR 60 TC 9 Z9 9 U1 0 U2 2 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 JUN PY 2002 VL 76 IS 12 BP 6016 EP 6026 DI 10.1128/JVI.76.12.6016-6026.2002 PG 11 WC Virology SC Virology GA 557MK UT WOS:000175912200018 PM 12021334 ER PT J AU Mizukoshi, E Nascimbeni, M Blaustein, JB Mihalik, K Rice, CM Liang, TJ Feinstone, SM Rehermann, B AF Mizukoshi, E Nascimbeni, M Blaustein, JB Mihalik, K Rice, CM Liang, TJ Feinstone, SM Rehermann, B TI Molecular and immunological significance of chimpanzee major histocompatibility complex haplotypes for hepatitis C virus immune response and vaccination studies SO JOURNAL OF VIROLOGY LA English DT Article ID MHC CLASS-I; T-LYMPHOCYTE RESPONSE; PEPTIDE-BINDING MOTIFS; HLA-A ALLELES; CELL EPITOPES; NUCLEOTIDE SUBSTITUTION; OVERDOMINANT SELECTION; HLA-A2.1 MOLECULES; INFECTION; RESPONSIVENESS AB The chimpanzee is a critical animal model for studying cellular immune responses to infectious pathogens such as hepatitis B and C viruses, human immunodeficiency virus, and malaria. Several candidate vaccines and immunotherapies for these infections aim at the induction or enhancement of cellular immune responses against viral epitopes presented by common human major his to compatibility complex (MHC) alleles. To identify and characterize chimpanzee MHC class I molecules that are functionally related to human alleles, we sequenced 18 different Pan troglodytes (Patr) alleles of 14 chimpanzees, 2 of them previously unknown and 3 with only partially reported sequences. Comparative analysis of Patr binding pockets and binding assays with biotinylated peptides demonstrated a molecular homology between the binding grooves of individual Patr alleles and the common human alleles HLA-A1, -A2, -A3, and -B7. Using cytotoxic T cells isolated from the blood of hepatitis C virus (HCV)-infected chimpanzees, we then mapped the Patr restriction of these HCV peptides and demonstrated functional homology between the Patr-HLA orthologues in cytotoxicity and gamma interferon (IFN-gamma) release assays. Based on these results, 21 HCV epitopes were selected to characterize the chimpanzees' cellular immune response to HCV. In each case, IFN-gamma-producing T cells were detectable in the blood after but not prior to HCV infection and were specifically targeted against those HCV peptides predicted by Patr-HLA homology. This study demonstrates a close functional homology between individual Patr and HLA alleles and shows that HCV infection generates HCV peptides that are recognized by both chimpanzees and humans with Patr and HLA orthologues. These results are relevant for the design and evaluation of vaccines in chimpanzees that can now be selected according to the most frequent human MHC haplotypes. C1 NIDDK, Liver Dis Sect, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Lab Hepatatis Res, Bethesda, MD 20892 USA. Rockefeller Univ, Ctr Study Hepatitis C, New York, NY 10021 USA. RP Rehermann, B (reprint author), NIDDK, Liver Dis Sect, NIH, 10 Ctr Dr,Room 9B16, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA85883-01, R01 CA085883] NR 54 TC 25 Z9 27 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 JUN PY 2002 VL 76 IS 12 BP 6093 EP 6103 DI 10.1128/JVI.76.12.6093-6103.2002 PG 11 WC Virology SC Virology GA 557MK UT WOS:000175912200026 PM 12021342 ER PT J AU Amara, RR Smith, JM Staprans, SI Montefiori, DC Villinger, F Altman, JD O'Neil, SP Kozyr, NL Xu, Y Wyatt, LS Earl, PL Herndon, JG McNicholl, JM McClure, HM Moss, B Robinson, HL AF Amara, RR Smith, JM Staprans, SI Montefiori, DC Villinger, F Altman, JD O'Neil, SP Kozyr, NL Xu, Y Wyatt, LS Earl, PL Herndon, JG McNicholl, JM McClure, HM Moss, B Robinson, HL TI Critical role for Env as well as Gag-Pol in control of a simian-human immunodeficiency virus 89.6P challenge by a DNA prime/recombinant modified vaccinia virus Ankara vaccine SO JOURNAL OF VIROLOGY LA English DT Article ID POLYMERASE CHAIN-REACTION; T-CELL DEPLETION; RHESUS-MONKEYS; NEUTRALIZING ANTIBODIES; MACAQUES; AIDS; INFECTION; ENVELOPE; PREVENTION; INDUCTION AB Cellular immune responses against epitopes in conserved Gag and Pol sequences of human immunodeficiency virus type 1 have become popular targets for candidate AIDS vaccines. Recently, we used a simian-human immunodeficiency virus model (SHIV 89.6P) with macaques to demonstrate the control of a pathogenic mucosal challenge by priming with Gag-Pol-Env-expressing DNA and boosting with Gag-Pol-Env-expressing recombinant modified vaccinia virus Ankara (rMVA). Here we tested Gag-Pol DNA priming and Gag-Pol rMVA boosting to evaluate the contribution of anti-Env immune responses to viral control. The Gag-Pol vaccine raised frequencies of Gag-specific T cells similar to those raised by the Gag-Pol-Env vaccine. Following challenge, these rapidly expanded to counter the challenge infection. Despite this, the control of the SHIV 89.6P challenge was delayed and inconsistent in the Gag-Pol-vaccinated group and all of the animals underwent severe and, in most cases, sustained loss of CD4(+) cells. Interestingly, most of the CD4(+) cells that were lost in the Gag-Pol-vaccinated group were uninfected cells. We suggest that the rapid appearance of binding antibody for Env in Gag-Pol-Env-vaccinated animals helped protect uninfected CD4(+) cells from Env-induced apoptosis. Our results highlight the importance of immune responses to Env, as well as to Gag-Pol, in the control of immunodeficiency virus challenges and the protection of CD4(+) cells. C1 Emory Univ, Sch Med, Vaccine Res Ctr, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Yerkes Reg Primate Res Ctr, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Pathol & Lab Med, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Microbiol & Immunol, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA. Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA. NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Div AIDS STD & TB Lab Res, Natl Ctr Infect Dis, Atlanta, GA 30330 USA. RP Robinson, HL (reprint author), 954 Gatewood Rd NE, Atlanta, GA 30329 USA. FU NCRR NIH HHS [P51 RR000165, P51 RR00165]; NIAID NIH HHS [AI 85343, P01 AI 43045, P01 AI043045]; NIDA NIH HHS [P30 DA 12121] NR 23 TC 136 Z9 147 U1 0 U2 2 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 JUN PY 2002 VL 76 IS 12 BP 6138 EP 6146 DI 10.1128/JVI.76.12.6138-6146.2002 PG 9 WC Virology SC Virology GA 557MK UT WOS:000175912200031 PM 12021347 ER PT J AU Teng, MN Collins, PL AF Teng, MN Collins, PL TI The central conserved cystine noose of the attachment G protein of human respiratory syncytial virus is not required for efficient viral infection in vitro or in vivo SO JOURNAL OF VIROLOGY LA English DT Article ID NECROSIS-FACTOR RECEPTOR; REPLICATION IN-VITRO; G-GLYCOPROTEIN; O-GLYCOSYLATION; SUBGROUP-B; SH-GENE; ESCAPE MUTANTS; F-PROTEIN; CELLS; HEPARIN AB The G glycoprotein of human respiratory syncytial virus (RSV) was identified previously as the viral attachment protein. Although we and others recently showed that G is not essential for replication in vitro, it does affect the efficiency of replication in a cell type-dependent fashion and is required for efficient replication in vivo. The ectodomain of G is composed of two heavily glycosylated domains with mucin-like characteristics that are separated by a short central region that is relatively devoid of glycosylation sites. This central region contains a 13-amino acid segment that is conserved in the same form among RSV isolates and is overlapped by a second segment containing four cysteine residues whose spacings are conserved in the same form and which create a cystine noose. The conserved nature of the cystine noose and flanking 13-amino acid segment suggested that this region likely was important for attachment activity. To test this hypothesis, we constructed recombinant RSVs from which the region containing the cysteine residues was deleted together with part or all of the conserved 13-amino acid segment. Surprisingly, each deletion had little or no effect on the intracellular synthesis and processing of the G protein, the kinetics or efficiency of virus replication in vitro, or sensitivity to neutralization by soluble heparin in vitro. In addition, neither deletion had any discernible effect on the ability of RSV to infect the upper respiratory tract of mice and both resulted in a 3- to 10-fold reduction in the lower respiratory tract. Thus, although the G protein is necessary for efficient virus replication in vivo, this activity does not require the central conserved cystine noose region. C1 NIAID, Infect Dis Lab, Bethesda, MD 20892 USA. RP Collins, PL (reprint author), NIAID, Infect Dis Lab, 50 S Dr,MSC 8007,Bldg 50,Room 6503, Bethesda, MD 20892 USA. RI Teng, Michael/I-5006-2012 OI Teng, Michael/0000-0002-0722-3659 NR 42 TC 42 Z9 44 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 JUN PY 2002 VL 76 IS 12 BP 6164 EP 6171 DI 10.1128/JVI.76.12.6164-6171.2002 PG 8 WC Virology SC Virology GA 557MK UT WOS:000175912200034 PM 12021350 ER PT J AU Santra, S Barouch, DH Kuroda, MJ Schmitz, JE Krivulka, GR Beaudry, K Lord, CI Lifton, MA Wyatt, LS Moss, B Hirsch, VM Letvin, NL AF Santra, S Barouch, DH Kuroda, MJ Schmitz, JE Krivulka, GR Beaudry, K Lord, CI Lifton, MA Wyatt, LS Moss, B Hirsch, VM Letvin, NL TI Prior vaccination increases the epitopic breadth of the cytotoxic T-lymphocyte response that evolves in rhesus monkeys following a simian-human immunodeficiency virus infection SO JOURNAL OF VIROLOGY LA English DT Article ID DISEASE PROGRESSION; DNA VACCINATION; VIREMIA; MACAQUES; COMPLEX; AIDS; INDUCTION; DOMINANT; REGIMEN; LOAD AB Although recent evidence has confirmed the importance of cytotoxic T-lymphocyte (CTL) responses in controlling human immunodeficiency virus type 1 and simian immunodeficiency virus replication, the relevance of the epitopic breadth of those CTL responses remains unexplored. In the present study, we sought to determine whether vaccination can expand CTL populations which recognize a repertoire of viral epitopes that is greater than is typically generated in the course of a viral infection. We demonstrate that potent secondary CTL responses to subdominant epitopes are rapidly generated following a pathogenic simian-human immunodeficiency virus challenge of rhesus monkeys vaccinated with plasmid DNA or recombinant modified vaccinia virus Ankara vaccines. These data indicate that prior vaccination can increase the breadth of the CTL response that evolves after an AIDS virus infection. C1 Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med,Div Viral Pathogenesis, Boston, MA 02215 USA. NIAID, Mol Microbiol Lab, NIH, Rockville, MD 20852 USA. NIAID, Viral Dis Lab, NIH, Rockville, MD 20852 USA. RP Letvin, NL (reprint author), Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med,Div Viral Pathogenesis, RE113,POB 15732, Boston, MA 02215 USA. FU NCI NIH HHS [CA-50139, R01 CA050139]; NIAID NIH HHS [AI-20729, R37 AI020729, R01 AI020729, U01 AI026507, AI-26507, AI-85343, P01 AI026507] NR 23 TC 18 Z9 19 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 JUN PY 2002 VL 76 IS 12 BP 6376 EP 6381 DI 10.1128/JVI.76.12.6376-6381.2002 PG 6 WC Virology SC Virology GA 557MK UT WOS:000175912200055 PM 12021371 ER PT J AU Smith, JR Venable, S Roberts, TW Metter, EJ Monticone, R Schneider, EL AF Smith, JR Venable, S Roberts, TW Metter, EJ Monticone, R Schneider, EL TI Relationship between in vivo age and in vitro aging: Assessment of 669 cell cultures derived from members of the Baltimore Longitudinal Study of Aging SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID REPLICATIVE LIFE-SPAN; SMOOTH-MUSCLE CELLS; SKIN FIBROBLASTS; DONOR AGE; CALORIC RESTRICTION; INVITRO; INVIVO; CAPACITY; GROWTH AB We examined the in vitro proliferative potential of 669 cell cultures established from skin biopsies of members of the Baltimore Longitudinal Study of Aging. The colony size distribution was used to estimate the proliferative life span of the cultures. A significant decline in proliferative potential with donor age was observed for female but not male donors. For both male and female donors, the proliferative potential was significantly greater for donors under the age of 30 years compared with all donors over the age of 30 years. In an attempt to reduce genetic heterogeneity, we examined the proliferative potential of cultures derived at different ages from the same donor. These studies revealed a trend (approaching statistical significance) toward low proliferative potential as donors aged. Interestingly, samples obtained from donors who had a history of skin cancer at the time of biopsy had a significantly lower doubling potential than those from donors who did not. The implications of these results for the use of cells derived from donors of different ages for aging research are discussed. C1 Baylor Coll Med, Huffington Ctr Aging, Houston, TX 77030 USA. Life Technol Inc, Gaithersburg, MD USA. NIA, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Univ So Calif, Ethel Percy Andrus Gerontol Ctr, Los Angeles, CA 90089 USA. RP Smith, JR (reprint author), Barshop Ctr Longev & Aging Studies, Dept Pathol, STCBM Bldg,15335 Lambda Dr, San Antonio, TX 78245 USA. NR 24 TC 23 Z9 26 U1 2 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 JUN PY 2002 VL 57 IS 6 BP B239 EP B246 PG 8 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 558RG UT WOS:000175979900004 PM 12023260 ER PT J AU Kobayashi, H Kawamoto, S Jo, SK Sato, N Saga, T Hiraga, A Konishi, J Hu, S Togashi, K Brechbiel, MW Star, RA AF Kobayashi, H Kawamoto, S Jo, SK Sato, N Saga, T Hiraga, A Konishi, J Hu, S Togashi, K Brechbiel, MW Star, RA TI Renal tubular damage detected by dynamic micro-MRI with a dendrimer-based magnetic resonance contrast agent SO KIDNEY INTERNATIONAL LA English DT Article DE proximal tubules; magnetic resonance imaging; dendrimer; contrast agent; cisplatin; non-invasive imaging ID POLYAMIDOAMINE DENDRIMER; CORE; NEPHROTOXICITY; ANGIOGRAPHY; FAILURE; DTPA AB Background. A noninvasive technique to evaluate the structure and function of the kidney would be useful to investigate renal diseases, especially acute renal failure. We have developed a novel technique to visualize functional micro-magnetic resonance (MR) images of the mouse kidney with a dendrimer-based macromolecular renal MR contrast agent. Method. Mice were injected with cisplatin or vehicle, then examined three days later by contrast-enhanced, dynamic high-resolution micro-MRI with 160 mum spatial resolution using a 1.5 T clinical MRI unit, a surface coil, and the renal contrast agent G4D-(1B4M-Gd)(64) . Results. The cortex and outer stripe of the outer medulla of the mouse kidney were clearly visualized in the normal mice. In animals treated with cisplatin, the gradation of tubular damage as assessed by contrast enhanced dynamic MRI correlated with renal function. Conclusion. Contrast-enhanced, dynamic high-resolution micro-MRI with a novel dendrimer-based macromolecular renal MR contrast agent can be a powerful tool for in vivo observation of renal structural and functional damage. C1 NCI, Metab Branch, Radioimmune & Inorgan Chem Sect, Radiat Oncol Branch,NIH, Bethesda, MD 20892 USA. Kyoto Univ, Dept Diagnost & Intervent Imagiol, Kyoto, Japan. Kyoto Univ, Dept Nucl Med & Diagnost Imaging, Kyoto, Japan. Kyoto Univ, Dept Radiol, Kyoto, Japan. NIDDKD, Renal Diagnost & Therapeut Unit, NIH, Bethesda, MD 20892 USA. RP Kobayashi, H (reprint author), NCI, Metab Branch, Radioimmune & Inorgan Chem Sect, Radiat Oncol Branch,NIH, Bldg 10,Room 4N109,10 Ctr Dr,MSC 1374, Bethesda, MD 20892 USA. NR 19 TC 64 Z9 66 U1 0 U2 1 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD JUN PY 2002 VL 61 IS 6 BP 1980 EP 1985 DI 10.1046/j.1523-1755.2002.00364.x PG 6 WC Urology & Nephrology SC Urology & Nephrology GA 553FX UT WOS:000175666100006 PM 12028438 ER PT J AU Bickel, CA Knepper, MA Verbalis, JG Ecelbarger, CA AF Bickel, CA Knepper, MA Verbalis, JG Ecelbarger, CA TI Dysregulation of renal salt and water transport proteins in diabetic Zucker rats SO KIDNEY INTERNATIONAL LA English DT Article; Proceedings Paper CT 33rd Annual Meeting of the American-Society-of-Nephrology CY OCT 10-16, 2000 CL TORONTO, CANADA SP Amer Soc Nephrol DE diabetic nephropathy; insulin; hypertension; Na, K-ATPase; Na-Cl cotransporter; epithelial sodium channel; aquaporin; Na-K-2Cl cotransporter; sodium transport ID THICK ASCENDING LIMB; NITRIC-OXIDE; VASOPRESSIN; NEPHROPATHY; CHANNEL; EXPRESSION; ABUNDANCE; MELLITUS; KIDNEY; MICE AB Background. Diabetic nephropathy is commonly associated with renal salt and water retention and hypertension. The molecular mechanisms involved and how the kidney responds to this volume expansion, in terms of renal transporter regulation, are not understood. Methods. Targeted proteomics employing semiquantitative immunoblotting were used to investigate regulation of abundance of the primary salt and water transport proteins of the kidney, in 6-month-old lean and obese Zucker rats, a model for Type II diabetes. Results. Obese rats were significantly heavier, had larger kidneys, increased plasma creatinine and glucose levels and elevated blood pressures. Furthermore, they had a marked decrease in abundance of many pre-macula densa renal sodium transporters. Mean band densities (% lean) were: in cortex, sodium phosphate cotransporter (NaPi-2), 68%, and sodium hydrogen exchanger (NHE3), 66%; and in outer medulla, NHE3, 39%, and the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2), 37%. Collecting duct proteins also were markedly reduced. In inner medulla, aquaporins-2, -3, and -4 were reduced to, 46, 48, and 46%, respectively, and the apical urea transporter, UTA1 to 52%. In contrast, post-macula densa sodium transporters were less affected. The thiazide-sensitive Na-Cl cotransporter (NCC) was 106% and the alpha- beta- and gamma-subunits of the epithelial sodium channel (ENaC), 54, 121, and 84% of lean, respectively. Conclusions. In obese rats, selective decreases for pre-macula densa sodium transporters may reflect decreased glomerular filtration rate and glomerulotubular balance. This potentially could reduce blood pressure by decreasing proximal tubule sodium reabsorption. C1 Georgetown Univ, Dept Med, Div Endocrinol & Metab, Washington, DC 20007 USA. NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. RP Ecelbarger, CA (reprint author), Georgetown Univ, Dept Med, Div Endocrinol & Metab, Bldg D,Rm 232,4000 Reservoir Rd NW, Washington, DC 20007 USA. FU Intramural NIH HHS [Z99 HL999999, Z01 HL001285-21]; NHLBI NIH HHS [R01 HL073193]; NIDDK NIH HHS [DK38094, K01 DK02672] NR 45 TC 43 Z9 46 U1 0 U2 0 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD JUN PY 2002 VL 61 IS 6 BP 2099 EP 2110 DI 10.1046/j.1523-1755.2002.00353.x PG 12 WC Urology & Nephrology SC Urology & Nephrology GA 553FX UT WOS:000175666100018 PM 12028450 ER PT J AU Kaysen, GA Dubin, JA Muller, HG Mitch, WE Rosales, LM Levin, NW AF Kaysen, GA Dubin, JA Muller, HG Mitch, WE Rosales, LM Levin, NW CA HEMO Study Group TI Relationships among inflammation nutrition and physiologic mechanisms establishing albumin levels in hemodialysis patients SO KIDNEY INTERNATIONAL LA English DT Article DE albumin synthesis; fractional catabolic rate; plasma volume; acute phase; C-reactive protein; ceruloplasmin; alpha 1 acid glycoprotein; nutrition; body mass index; nPCR ID ACUTE-PHASE PROTEIN; CHRONIC-RENAL-FAILURE; PLASMA-ALBUMIN; SERUM-ALBUMIN; MORTALITY; LIVER; HYPOALBUMINEMIA; MALNUTRITION; FIBRINOGEN; KINETICS AB Background. Serum albumin concentration is a balance among its synthesis rate, fractional catabolic rate (FCR), distribution, dilution in the plasma pool and external loss. The physiologic bases for establishing the level of serum albumin in hemodialysis patients have not been defined despite the association of hypoalbuminemia with excess mortality. Albumin concentration is associated with the levels of several acute phase proteins (APPs), C-reactive protein (CRP), alpha1 acid glycoprotein (alpha1 AG), or ceruloplasmin, and with nutritional markers, such as normalized protein catabolic rate (nPCR). Methods. To establish the relationship among parameters that regulate albumin levels and markers of nutrition and inflammation, we injected [(125) I]-albumin, into 64 hemodialysis patients enrolled in the HEMO study to measure albumin distribution, synthesis and FCR. These variables were related to the levels of acute phase proteins (APPs), nPCR, body mass index (BMI), external albumin loss as well as demographic variables. Albumin distribution, synthesis and FCR were calculated from kinetic modeling, as was the initial plasma volume (PV). Serum albumin, transferrin, CRP, ceruloplasmin and alpha1 AG were measured weekly. Dialysate was collected during one dialysis each week to measure albumin loss. Results were analyzed by multiple linear regression. Results. Albumin concentration correlated with its synthesis rate and FCR, but not with PV or its distribution between the vascular and extravascular pools. Albumin concentration also correlated with nPCR and alpha1 AG. However, albumin synthesis was directly related most strongly to PV and BMI (or nPCR), but not to levels of APPs. By contrast, albumin FCR correlated positively with both alpha1 AG and ceruloplasmin. Conclusion. Albumin concentration in dialysis patients changes with inflammation and nutritional status through their effects on albumin catabolism and synthesis, respectively. Within the range of albumin levels in these patients, nutritional variables primarily affected albumin synthesis while inflammation caused hypoalbuminemia by increasing albumin FCR. Albumin synthesis also increased in proportion to PV. The result of this is that PV expansion does not contribute to hypoalbuminemia. C1 Univ Calif Davis, Div Nephrol, Dept Med, Davis, CA 95616 USA. Univ Calif Davis, Dept Stat, Davis, CA 95616 USA. VANCHCS, Res Serv, Mather, CA USA. Emory Univ, Dept Med, Div Nephrol, Atlanta, GA 30322 USA. Renal Res Inst New York, New York, NY USA. NIDDK, HEMO Study Grp, NIH, Bethesda, MD USA. Yale Univ, Div Biostat, New Haven, CT USA. RP Kaysen, GA (reprint author), Univ Calif Davis, Div Nephrol, Dept Med, TB 136, Davis, CA 95616 USA. FU NCRR NIH HHS [M01-RR00039]; NIDDK NIH HHS [R01 DK 50777] NR 38 TC 139 Z9 147 U1 0 U2 3 PU BLACKWELL PUBLISHING INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD JUN PY 2002 VL 61 IS 6 BP 2240 EP 2249 DI 10.1046/j.1523-1755.2002.00076.x PG 10 WC Urology & Nephrology SC Urology & Nephrology GA 553FX UT WOS:000175666100034 PM 12028466 ER PT J AU Carney, SA Tahara, H Swartz, CD Risinger, JI He, H Moore, AB Haseman, JK Barrett, JC Dixon, D AF Carney, SA Tahara, H Swartz, CD Risinger, JI He, H Moore, AB Haseman, JK Barrett, JC Dixon, D TI Immortalization of human uterine leiomyoma and myometrial cell lines after induction of telomerase activity: Molecular and phenotypic characteristics SO LABORATORY INVESTIGATION LA English DT Article ID REVERSE-TRANSCRIPTASE HTERT; CATALYTIC SUBUNIT; GROWTH-FACTOR; TUMOR-CELLS; INVITRO; CANCER; PROGESTERONE; FIBROBLASTS; EXPRESSION; ESTROGEN AB in vitro model systems for studying uterine leiomyomas are limited in that human-derived leiomyoma cells grow poorly in culture compared with normal myometrial cells and begin to senesce early, at approximately passage 10 in our studies. To our knowledge, a good in vitro human-derived cell culturing system for leiomyomas does not exist. In an attempt to fill this void, we have immortalized a uterine leiomyoma cell line by inducing telomerase activity, which allows cells to bypass their normal programmed senescence. Telomerase activity was induced by infecting the target (uterine leiomyoma and normal myometrial) cells with a retroviral vector containing hTERT, the gene for the catalytic subunit of telomerase. Subsequent analysis by RT-PCR and the telomeric repeat amplification protocol assay confirmed expression of the inserted gene and induction of telomerase activity in leiomyoma and myometrial cells. Analysis of cells for estrogen receptor-alpha and progesterone receptor proteins by Western blotting showed no change in expression of these proteins between the immortalized and parental leiomyoma and myometrial cells. Both immortalized and parental myometrial and leiomyoma cells expressed the smooth muscle-specific cytoskeletal protein alpha-actin and were negative for mutant p53 protein as evidenced by immunocytochemical staining. The immortalized leiomyoma and myometrial cells showed no anchorage-independent growth, with the exception of a small subpopulation of immortalized leiomyoma cells at a higher passage that did form two to three small colonies (per 50,000 cells) in soft agar. None of the immortalized cells were tumorigenic in nude mice. In conclusion, our data show the successful insertion of the hTERT gene into leiomyoma and myometrial cells and the immortalization of these cell lines without phenotypic alteration from the parental cell types (up to 200 population doublings). These cells should help to advance research in understanding the molecular pathways involved in the conversion of a normal myometrial cell to a leiomyoma cell and the mechanisms responsible for the growth of uterine leiomyomas. Answers to these questions will undoubtedly lead to the development of more effective treatment and intervention regimens for clinical cases of uterine leiomyoma. C1 NIEHS, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. NCI, Lab Biosyst & Canc, Bethesda, MD 20892 USA. RP Dixon, D (reprint author), NIEHS, Lab Expt Pathol, POB 12233,MD C2-09, Res Triangle Pk, NC 27709 USA. NR 30 TC 74 Z9 77 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0023-6837 J9 LAB INVEST JI Lab. Invest. PD JUN PY 2002 VL 82 IS 6 BP 719 EP 728 DI 10.1097/01.LAB.0000017499.51216.3E PG 10 WC Medicine, Research & Experimental; Pathology SC Research & Experimental Medicine; Pathology GA 564MF UT WOS:000176317600006 PM 12065682 ER PT J AU Rapoport, SI AF Rapoport, SI TI Hydrogen magnetic resonance spectroscopy in Alzheimer's disease SO LANCET NEUROLOGY LA English DT Letter ID H-1 MRS C1 NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. RP Rapoport, SI (reprint author), NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. NR 5 TC 6 Z9 7 U1 1 U2 1 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 1474-4422 J9 LANCET NEUROL JI Lancet Neurol. PD JUN PY 2002 VL 1 IS 2 BP 82 EP 82 DI 10.1016/S1474-4422(02)00035-2 PG 1 WC Clinical Neurology SC Neurosciences & Neurology GA 588HA UT WOS:000177695000014 PM 12849509 ER PT J AU Bodurka, J Bandettini, PA AF Bodurka, J Bandettini, PA TI Toward direct mapping of neuronal activity: MRI detection of ultraweak, transient magnetic fields changes SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE magnetic field; magnetic phase; neuronal activity; MRI; brain mapping ID HUMAN BRAIN; RESONANCE; EPI; MAGNETOENCEPHALOGRAPHY; NETWORKS; TESLA AB A novel method based on selective detection of rapidly changing DeltaB(o), magnetic fields and suppression of slowly changing DeltaB(o), fields is presented. The ultimate goal of this work is to present a method that may allow detection of transient and subtle changes in B-o in cortical tissue associated with electrical currents produced by neuronal activity. The method involves the detection of NMR phase changes that occur during a single-shot spin-echo (SE) echo-planar sequence (EPI) echo time. SE EPI effectively rephases all changes in B-o that occur on a time scale longer than the echo time (TE) and amplifies all DeltaB(o) changes that occur during TE/2. The method was tested on a phantom that contains wires in which current can be modulated. The sensitivity and flexibility of the technique was demonstrated by modulation of the temporal position and duration of the stimuli-evoked transient magnetic field relative to the 180 RF pulse in the imaging sequence-requiring precise stimulus timing. Currently, with this method magnetic field changes as small as 2 x 10(-10) T (200 pT) and lasting for 40 msec can be detected. Implications for direct mapping of brain neuronal activity with MRI are discussed. C1 NIMH, Telsa Funct Neuroimaging Facil 3, NIH, Bethesda, MD 20892 USA. NIMH, Lab Brain & Cognit, Unit Funct Imaging Methods, NIH, Bethesda, MD 20892 USA. RP Bodurka, J (reprint author), NIMH, Telsa Funct Neuroimaging Facil 3, NIH, Bldg 10,Rm 1D80,10 Ctr Dr,MSC 1148, Bethesda, MD 20892 USA. NR 27 TC 120 Z9 122 U1 0 U2 9 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD JUN PY 2002 VL 47 IS 6 BP 1052 EP 1058 DI 10.1002/mrm.10159 PG 7 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 557XV UT WOS:000175935100002 PM 12111950 ER PT J AU Barbier, EL St Lawrence, KS Grillon, E Koretsky, AP Decorps, M AF Barbier, EL St Lawrence, KS Grillon, E Koretsky, AP Decorps, M TI A model of blood-brain barrier permeability to water: Accounting for blood inflow and longitudinal relaxation effects SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE contrast agent; cerebral blood flow; cerebral blood volume; blood-brain barrier; permeability; water ID SUSCEPTIBILITY-CONTRAST MRI; MAGNETIC-RESONANCE; CEREBRAL-CORTEX; VOLUME; FLOW; OXYGENATION; PERFUSION; ISCHEMIA; INVIVO AB A noninvasive technique for measuring the permeability of the blood-brain barrier (BBB) to water could help to evaluate changes in the functional integrity of the BBB that occur in different pathologies, such as multiple sclerosis or growth of brain tumor. Recently, Schwarzbauer et al. (Magn Reson Med 1997;37:769-777) proposed an MR method to measure this permeability based on the T-1 reductions induced by injecting various doses of paramagnetic contrast agent. However, this method may be difficult to implement in a clinical environment. Described here is a two-point technique, in which a spatially selective inversion is used to measure T-1 prior to and after injection of an intravascular contrast agent. Measurements made in the rat brain are compared to numerical simulations generated with a physiological model that accounts for blood flow and includes two different blood volumes: nonexchanging and exchanging blood volumes. Our results suggest that BBB permeability could be evaluated from the change in T-1 caused by the vascular contrast agent. This technique might provide an approach for monitoring changes in BBB permeability to water in clinical studies. C1 INSERM, U438, RMN Bioclin, Grenoble, France. NINCDS, Lab Funct & Mol Imaging, NIH, Bethesda, MD 20892 USA. NIH, Ctr Clin, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. RP Barbier, EL (reprint author), CHU Michallon Pavillon B, RMN Bioclin, INSERM, U438, F-38043 Grenoble 9, France. RI Barbier, Emmanuel/E-6302-2011; St. Lawrence, Keith/B-5726-2015; Koretsky, Alan/C-7940-2015 OI Barbier, Emmanuel/0000-0002-4952-1240; Koretsky, Alan/0000-0002-8085-4756 NR 30 TC 13 Z9 13 U1 0 U2 2 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD JUN PY 2002 VL 47 IS 6 BP 1100 EP 1109 DI 10.1002/mrm.10158 PG 10 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 557XV UT WOS:000175935100008 PM 12111956 ER PT J AU de Zwart, JA Ledden, PJ Kellman, P van Gelderen, P Duyn, JH AF de Zwart, JA Ledden, PJ Kellman, P van Gelderen, P Duyn, JH TI Design of a SENSE-optimized high-sensitivity MRI receive coil for brain imaging SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE MRI; parallel imaging; SENSE; brain imaging; RF coil; sensitivity; SNR ID RADIOFREQUENCY COIL; PHASED-ARRAY; HEAD-COIL; NMR AB An 8-channel receive-only detector array was developed for SENSE MRI of human brain. The coil geometry was based on a gapped element design and used ultra-high impedance preamplifiers for mutual decoupling of the elements. Computer simulations of the electric and magnetic fields showed that excellent signal-to-noise ratio (SNR) and SENSE performance could be achieved by placing the coil elements close to the head and maintaining a substantial gap between the elements. Measurements with a 1.5 T prototype coil showed a 2.7-fold improvement of the SNR averaged over the brain compared to a conventional quadrature birdcage receive coil and an average geometrical noise amplification factor (g-value) of 1.06 and 1.38 for rate-2 and rate-3 SENSE, respectively. C1 NINCDS, LFMI, NIH, Bethesda, MD 20892 USA. Nova Med, Wakefield, MA USA. NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Duyn, JH (reprint author), NINCDS, LFMI, NIH, Bldg 10,Rm B1D-118,MSC 1065,10 Ctr Dr, Bethesda, MD 20892 USA. RI Duyn, Jozef/F-2483-2010 NR 16 TC 110 Z9 111 U1 1 U2 5 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD JUN PY 2002 VL 47 IS 6 BP 1218 EP 1227 DI 10.1002/mrm.10169 PG 10 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 557XV UT WOS:000175935100021 PM 12111969 ER PT J AU Park, YG Lukes, L Yang, H Debies, MT Samant, RS Welch, DR Lee, M Hunter, KW AF Park, YG Lukes, L Yang, H Debies, MT Samant, RS Welch, DR Lee, M Hunter, KW TI Comparative sequence analysis in eight inbred strains of the metastasis modifier QTL candidate gene Brms1 SO MAMMALIAN GENOME LA English DT Article ID MOUSE; REGIONS C1 NCI, DCEG, Lab Populat Genet, NIH, Bethesda, MD 20892 USA. Penn State Univ, Coll Med, Dept Med, Jake Gittlen Canc Res Inst, Hershey, PA USA. RP Hunter, KW (reprint author), NCI, DCEG, Lab Populat Genet, NIH, Bldg 41,Room D702,41 Ctr Dr, Bethesda, MD 20892 USA. RI Welch, Danny/B-7310-2009 OI Welch, Danny/0000-0002-1951-4947 NR 10 TC 12 Z9 12 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 JUN PY 2002 VL 13 IS 6 BP 289 EP 292 DI 10.1007/s00335-001-2151-6 PG 4 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 561BK UT WOS:000176116200002 PM 12115030 ER PT J AU Remmers, EF Hansen, CT Hashiramoto, A Wilder, RL Dobbins, DE AF Remmers, EF Hansen, CT Hashiramoto, A Wilder, RL Dobbins, DE TI Application of interval haplotype analysis facilitates efficient mapping of the mutation causing osteopetrosis in tl rats SO MAMMALIAN GENOME LA English DT Article ID MACROPHAGE; TOOTHLESS C1 NIAMSD, Arthritis & Rheumatism Branch, Inflammatory Joint Dis Sect, NIH, Bethesda, MD 20892 USA. NIH, Off Res Serv, Vet Resources Program, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Dept Anat Physiol & Genet, Bethesda, MD 20814 USA. RP Remmers, EF (reprint author), NIDDKD, NIH, 9000 Rockville Pike,Bldg 10,Room 9N308, Bethesda, MD 20892 USA. NR 8 TC 1 Z9 1 U1 0 U2 1 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 JUN PY 2002 VL 13 IS 6 BP 299 EP 301 DI 10.1007/s00335-001-1022-5 PG 3 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 561BK UT WOS:000176116200004 PM 12115032 ER PT J AU Lipscomb, J Snyder, CF AF Lipscomb, J Snyder, CF TI The outcomes of cancer outcomes research - Focusing on the National Cancer Institute's quality-of-care initiative SO MEDICAL CARE LA English DT Article DE outcomes research; quality of care; neoplasms ID PROSTATE-CANCER; PATTERNS AB BACKGROUND. To assess the impact of outcomes research, the Agency for Healthcare Policy and Research has proposed a four-level typology that successively distinguishes between research that (1) adds to the knowledge base only, (2) affects practice policies, (3) influences the delivery of care, and (4) leads to changes in health outcomes. The agency has concluded that outcomes research to date has made substantial contributions at level 1, but has had little impact at levels 2 to 4. OBJECTIVE. To describe the National Cancer Institute's emerging agenda in cancer outcomes research, focusing on how it has been shaped early on to support the institute's quality of care initiative and noting whether each new project represents outcomes research at level 1, 2, 3, or 4. RESULTS. Cancer outcomes research describes, interprets, and predicts the impact of various influences, especially (but not exclusively) interventions on endpoints that matter to decision makers. The institute's projects to improve the scientific quality of final endpoint measures, strengthen the methodologic and empirical basis for quality of care analysis, and enhance the application of health-related quality-of-life measures in clinical trials represent level 1 applications of outcomes research. Collaborative projects with federal agencies to translate evidence into practice and a new public and private effort to identify core measures of cancer quality represent level 2 and level 3 outcomes research. CONCLUSION. The stage has been set for carrying out a balanced portfolio of level 1 to 3 outcomes research projects in cancer, with the ultimate (level 4) aim of improving cancer-related health outcomes. C1 NCI, Div Canc Control & Populat Sci, Appl Res Program, Outcomes Res Branch, Bethesda, MD 20892 USA. RP Lipscomb, J (reprint author), NCI, Div Canc Control & Populat Sci, Appl Res Program, Outcomes Res Branch, 6130 Execut Blvd,MSC 7344,EPN Room 4005, Bethesda, MD 20892 USA. NR 15 TC 16 Z9 16 U1 1 U2 3 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 JUN PY 2002 VL 40 IS 6 SU S BP 3 EP 10 PG 8 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 556YG UT WOS:000175876700002 ER PT J AU Etzioni, R Riley, GF Ramsey, SD Brown, M AF Etzioni, R Riley, GF Ramsey, SD Brown, M TI Measuring costs - Administrative claims data, clinical trials, and beyond SO MEDICAL CARE LA English DT Article DE medical care costs; survival; cost-effectiveness analysis ID COLORECTAL-CANCER; MEDICAL COSTS; CARE; SURVIVAL; STAGE AB BACKGROUND. Accurate estimation of medical care costs raises a host of issues, both practical and methodological. OBJECTIVEs. This article reviews methods for estimating the long-term medical care costs associated with a cancer diagnosis. METHODS. The authors consider data from administrative claims databases and describe the analytic challenges posed by these increasingly common resources. They present a number of statistical methods that are valid under censoring and describe methods for estimating mean costs and controlling for covariates. In addition, the authors compare two different approaches for estimating the cancer-related costs; namely, the portion of the long-term costs that may be attributed to the disease. Examples from economic studies of breast and colorectal cancer are presented. RESULTS. In an analysis of data on colorectal cancer costs from the SEER-Medicare database, the two methods used to estimate expected long-term costs (one model based, one not model-based) yielded similar results. However, in calculating expected cancer-related costs, a method that included future medical costs among controls yielded quite different results from the method that did not include these future costs. CONCLUSIONS. Statistical methods for analyzing long-term medical costs under censoring are available and appropriate in many applications where total or disease-related costs are of interest. Several of these approaches are non-parametric and therefore may be expected to be robust against the non-standard features that are often encountered when analyzing medical cost data. C1 Fred Hutchinson Canc Res Ctr, Publ Hlth Sci Div, Seattle, WA USA. US Hlth Care Financing Adm, Baltimore, MD 21207 USA. NCI, Bethesda, MD 20892 USA. RP Etzioni, R (reprint author), Fred Hutchinson Canc Res Ctr, Publ Hlth Sci Div, 1100 Fairview Ave N,Box 19024,MP 665, Seattle, WA USA. NR 26 TC 17 Z9 17 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7079 J9 MED CARE JI Med. Care PD JUN PY 2002 VL 40 IS 6 SU S BP 63 EP 72 PG 10 WC Health Care Sciences & Services; Health Policy & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 556YG UT WOS:000175876700010 ER PT J AU Deye, J AF Deye, J TI The NCI and NIBIB impact on medical physics SO MEDICAL PHYSICS LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC PHYSICISTS MEDICINE AMER INST PHYSICS PI MELVILLE PA STE 1 NO 1, 2 HUNTINGTON QUADRANGLE, MELVILLE, NY 11747-4502 USA SN 0094-2405 J9 MED PHYS JI Med. Phys. PD JUN PY 2002 VL 29 IS 6 BP 1331 EP 1331 PG 1 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 565ME UT WOS:000176373400656 ER PT J AU Ahvazi, B Boeshans, K Jang, SI Kalinin, AE Steinert, PM AF Ahvazi, B Boeshans, K Jang, SI Kalinin, AE Steinert, PM TI Transglutaminases in skin biology SO MINERVA BIOTECNOLOGICA LA English DT Article; Proceedings Paper CT 7th International Conference on Transglutaminases and Protein Crosslinking Recactions CY SEP 14-17, 2002 CL FERRRA, ITALY DE protein glutamine gamma glutamyltransferase; genetics; skin; protein glutamine gamma glutamyltransferase; physiopathology ID CELL-ENVELOPE PROTEINS; SUBSTRATE PROPERTIES; LAMELLAR ICHTHYOSIS; CROSS-LINKING; GENE; FAMILY; MUTATIONS; LORICRIN AB During their terminal differentiation programs, all stratified squamous epithelia assemble an elaborate insoluble barrier structure, generically termed the cornified cell envelope (CE), on the inside of their plasma membrane at the cell periphery. This structure provides a vital physical and/or biomechanical barrier for the mature tissue and organism. The CE is composed of several defined structural proteins that become cross-linked together by the action of one or more transglutaminase (TGase) enzymes. Recent studies on the complexity of the family of TGase enzymes and the initial draft of the human genome have revealed the presence of nine TGase-like genes. Of these, six active genes/enzymes are now known to he expressed in epithelial cells. This bewildering complexity strongly implies that the proper assembly of the CE is indeed vital for survival. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. NR 25 TC 3 Z9 3 U1 0 U2 0 PU EDIZIONI MINERVA MEDICA PI TURIN PA CORSO BRAMANTE 83-85 INT JOURNALS DEPT., 10126 TURIN, ITALY SN 1120-4826 J9 MINERVA BIOTECNOL JI Minerva Biotechnol. PD JUN PY 2002 VL 14 IS 2 BP 165 EP 169 PG 5 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA 605PB UT WOS:000178685300010 ER PT J AU Davis-Hayman, SR Nash, TE AF Davis-Hayman, SR Nash, TE TI Review: Genetic transformation of parasitic organisms. 1 - Genetic manipulation of Giardia lamblia SO MOLECULAR AND BIOCHEMICAL PARASITOLOGY LA English DT Review DE Giardia lamblia; protozoa; transfection; genetic manipulation; gene regulation ID FIREFLY LUCIFERASE; RIBOSOMAL-RNA; EXPRESSION; VIRUS; PROMOTER; PROTEIN; IDENTIFICATION; TRANSFECTION; GENOME; TRANSLATION AB Giardia lamblia is a flagellated protozoan that infects several species including humans and is a major agent of waterborne outbreaks of diarrhea. G. lamblia is also important in the study of basic eukaryotic molecular biology and evolution, however, it has been difficult to employ standard genetic methods in the study of Giardia. Over the past 6 years. two transfection systems were developed and used for the genetic manipulation of G. lamblia. Both systems allow transient or stable transfection of Giardia and/or foreign genes. The DNA-based transfection system allows electroporation of circular or linear plasmid DNA into trophozoites. The RNA virus-based transfection system requires electroporation of in vitro transcribed RNA into GLV-infected trophozoites. Because G. lamblia is one of the most rudimentary eukaryotes, its processes of transcription. translation and protein transport. as well as its metabolic and biochemical pathways, are of interest. Study of these areas will continue to be advanced using transfection in combination With Cellular and molecular tools. Several groups have combined these technologies with other techniques to study protein transport and the transcriptional and post-transcriptional regulation of Giardia genes, including encystation-specific and variant Surface protein genes. In addition, coupling antisense techniques with transfection has permitted functional knockout of Giardia metabolic genes, allowing Giardia metabolic pathways to be Studied. In the near future, both transfection systems will be potent tools in our investigations of the perplexing questions in Giardia biology. (C) 2002 Published by Elsevier Science B.V. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Davis-Hayman, SR (reprint author), Room B1-06,Bldg 4,4 Ctr Dr,MSC 0425, Bethesda, MD 20892 USA. NR 31 TC 34 Z9 41 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-6851 J9 MOL BIOCHEM PARASIT JI Mol. Biochem. Parasitol. PD JUN PY 2002 VL 122 IS 1 BP 1 EP 7 AR PII S0166-6851(02)00063-4 DI 10.1016/S0166-6851(02)00063-4 PG 7 WC Biochemistry & Molecular Biology; Parasitology SC Biochemistry & Molecular Biology; Parasitology GA 581KG UT WOS:000177290900001 PM 12076765 ER PT J AU Hatfield, DL Gladyshev, VN AF Hatfield, DL Gladyshev, VN TI How selenium has altered our understanding of the genetic code SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Review ID SELENOCYSTEINE TRANSFER-RNA; SELENOPROTEIN MESSENGER-RNAS; HYDROPEROXIDE GLUTATHIONE-PEROXIDASE; MAMMALIAN THIOREDOXIN REDUCTASE; NONSENSE-MEDIATED DECAY; SERINE TRANSFER-RNA; DIETARY SELENIUM; ACTIVE-SITE; RAT-LIVER; FUNCTIONAL-CHARACTERIZATION C1 NCI, Mol Biol Selenium Sect, Basic Res Lab, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. Univ Nebraska, Dept Biochem, Lincoln, NE 68588 USA. RP Hatfield, DL (reprint author), NCI, Mol Biol Selenium Sect, Basic Res Lab, Ctr Canc Res,NIH, Bldg 37,Room 2D09, Bethesda, MD 20892 USA. RI Gladyshev, Vadim/A-9894-2013 FU NCI NIH HHS [CA061603, R01 CA080946]; NIGMS NIH HHS [GM080946, R01 GM061603] NR 103 TC 396 Z9 411 U1 5 U2 35 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 JUN PY 2002 VL 22 IS 11 BP 3565 EP 3576 DI 10.1128/MCB.22.11.3565-3576.2002 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 551JP UT WOS:000175557600001 PM 11997494 ER PT J AU Schwimmer, C Masison, DC AF Schwimmer, C Masison, DC TI Antagonistic interactions between yeast [PSI+] and [URE3] prions and curing of [URE3] by Hsp70 protein chaperone Ssa1p but not by Ssa2p SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID SACCHAROMYCES-CEREVISIAE; SUP35 PROTEIN; IN-VITRO; PROPAGATION; GENE; DETERMINANT; REPLICATION; HSP104; PSI(+); APPEARANCE AB The yeast [PSI+], [URE3], and [PIN+] genetic elements are prion forms of Sup35p, Ure2p, and Rnq1p, respectively. Overexpression of Sup35p, Ure2p, or Rnq1p leads to increased de novo appearance of [PSI+], [URE3], and [PIN+], respectively. This inducible appearance of [PSI+] was shown to be dependent on the presence of [PIN+] or [URE3] or overexpression of other yeast proteins that have stretches of polar residues similar to the prion-determining domains of the known prion proteins. In a similar manner, [PSI+] and [URE3] facilitate the appearance of [PIN+]. In contrast to these positive interactions, here we find that in the presence of [PIN+], IPSI+] and [URE3] repressed each other's propagation and de novo appearance. Elevated expression of Hsp104 and Hsp70 (Ssa2p) had little effect on these interactions, ruling out competition between the two prions for limiting amounts of these protein chaperones. In contrast, we find that constitutive overexpression of SSA1 but not SSA2 cured cells of [URE3], uncovering a specific interaction between Ssa1p and [URE3] and a functional distinction between these nearly identical Hsp70 isoforms. We also find that Hsp104 abundance, which critically affects [PSI+] propagation, is elevated when [URE3] is present. Our results are consistent with the notion that proteins that have a propensity to form prions may interact with heterologous prions but, as we now show, in a negative manner. Our data also suggest that differences in how [PSI+] and [URE3] interact with Hsp104 and Hsp70 may contribute to their antagonistic interactions. C1 NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Masison, DC (reprint author), NIDDKD, Lab Biochem & Genet, NIH, Bldg 8,Room 407, Bethesda, MD 20892 USA. NR 39 TC 122 Z9 124 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 JUN PY 2002 VL 22 IS 11 BP 3590 EP 3598 DI 10.1128/MCB.22.11.3590-3598.2002 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 551JP UT WOS:000175557600003 PM 11997496 ER PT J AU Fernandez-Salas, E Suh, KS Speransky, VV Bowers, WL Levy, JM Adams, T Pathak, KR Edwards, LE Hayes, DD Cheng, C Steven, AC Weinberg, WC Yuspa, SH AF Fernandez-Salas, E Suh, KS Speransky, VV Bowers, WL Levy, JM Adams, T Pathak, KR Edwards, LE Hayes, DD Cheng, C Steven, AC Weinberg, WC Yuspa, SH TI mtCLIC/CLIC4, an organellular chloride channel protein, is increased by DNA damage and participates in the apoptotic response to p53 SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID PROGRAMMED CELL-DEATH; MOLECULAR-CLONING; SUBCELLULAR-DISTRIBUTION; P53-INDUCED APOPTOSIS; POTENTIAL MEDIATOR; FAMILY; GENE; EXPRESSION; MEMBER; P64 AB mtCLIC/CLIC4 (referred to here as mtCLIC) is a p53- and tumor necrosis factor alpha-regulated cytoplasmic and mitochondrial protein that belongs to the CLIC family of intracellular chloride channels. mtCLIC associates with the inner mitochondrial membrane. Dual regulation of mtCLIC by two stress response pathways suggested that this chloride channel protein might contribute to the cellular response to cytotoxic stimuli. DNA damage or overexpression of p53 upregulates mtCLIC and induces apoptosis. Overexpression of mtCLIC by transient transfection reduces mitochondrial membrane potential, releases cytochrome c into the cytoplasm, activates caspases, and induces apoptosis. mtCLIC is additive with Bax in inducing apoptosis without a physical association of the two proteins. Antisense mtCLIC prevents the increase in mtCLIC levels and reduces apoptosis induced by p53 but not apoptosis induced by Bax, suggesting that the two proapoptotic proteins function through independent pathways. Our studies indicate that mtCLIC, like Bax, Noxa, p53AIP1, and PUMA, participates in a stress-induced death pathway converging on mitochondria and should be considered a target for cancer therapy through genetic or pharmacologic approaches. C1 NCI, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Immunobiol Lab, Bethesda, MD 20892 USA. RP Yuspa, SH (reprint author), NCI, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. RI Weinberg, Wendy/A-8920-2009 NR 58 TC 103 Z9 108 U1 0 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 JUN PY 2002 VL 22 IS 11 BP 3610 EP 3620 DI 10.1128/MCB.22.11.3610-3620.2002 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 551JP UT WOS:000175557600005 PM 11997498 ER PT J AU Farrell, CM West, AG Felsenfeld, G AF Farrell, CM West, AG Felsenfeld, G TI Conserved CTCF insulator elements flank the mouse and human beta-globin loci SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID OPEN CHROMATIN STRUCTURE; HUMAN ERYTHROID-CELLS; DNASE-I SENSITIVITY; CONTROL REGION; HYPERSENSITIVE SITES; GENE-EXPRESSION; RECEPTOR GENES; INTERGENIC TRANSCRIPTION; NUCLEAR-LOCALIZATION; HISTONE ACETYLATION AB A binding site for the transcription factor CTCF is responsible for enhancer-blocking activity in a variety of vertebrate insulators, including the insulators at the 5' and 3' chromatin boundaries of the chicken beta-globin locus. To date, no functional domain boundaries have been defined at mammalian beta-globin loci, which are embedded within arrays of functional olfactory receptor genes. In an attempt to define boundary elements that could separate these gene clusters, CTCF-binding sites were searched for at the most distal DNase 1-hypersensitive sites (HSs) of the mouse and human beta-globin loci. Conserved CTCF sites were found at 5'HS5 and 3'HS1 of both loci. All of these sites could bind to CTCF in vitro. The sites also functioned as insulators in enhancer-blocking assays at levels correlating with CTCF-binding affinity, although enhancer-blocking activity was weak with the mouse 5'HS5 site. These results show that with respect to enhancer-blocking elements, the architecture of the mouse and human beta-globin loci is similar to that found previously for the chicken beta-globin locus. Unlike the chicken locus, the mouse and human beta-globin loci do not have nearby transitions in chromatin structure but the data suggest that 3'HS1 and 5'HS5 may function as insulators that prevent inappropriate interactions between beta-globin regulatory elements and those of neighboring domains or subdomains, many of which possess strong enhancers. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Felsenfeld, G (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. OI West, Adam/0000-0003-3502-7804 NR 43 TC 125 Z9 131 U1 1 U2 5 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 JUN PY 2002 VL 22 IS 11 BP 3820 EP 3831 DI 10.1128/MCB.22.11.3820-3831.2002 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 551JP UT WOS:000175557600023 PM 11997516 ER PT J AU Hsia, SCV Shi, YB AF Hsia, SCV Shi, YB TI Chromatin disruption and histone acetylation in regulation of the human immunodeficiency virus type 1 long terminal repeat by thyroid hormone receptor SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID NUCLEOSOME REMODELING FACTOR; NUCLEAR RECEPTOR; TRANSCRIPTIONAL REPRESSION; N-COR; GLUCOCORTICOID RECEPTOR; MOLECULAR MECHANISMS; RESPONSE ELEMENT; SWI/SNF COMPLEX; BINDING PROTEIN; GENE-REGULATION AB The human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) controls the expression of HIV-1 viral genes and thus viral propagation and pathology. Numerous host factors participate in the regulation of the LTR promoter, including thyroid hormone (T-3) receptor (TR). In vitro, TR can bind to the promoter region containing the NF-kappaB and Sp1 binding sites. Using the frog oocyte as a model system for chromatin assembly mimicking that in somatic cells, we demonstrated that TR alone and TR/RXR (9-cis retinoic acid receptor) can bind to the LTR in vivo independently of T-3. Consistent with their ability to bind the LTR, both TR and TR/RXR can regulate LTR activity in vivo. In addition, our analysis of the plasmid minichromosome shows that T-3-bound TR disrupts the normal nucleosomal array structure. Chromatin immunoprecipitation assays with anti-acetylated-histone antibodies revealed that unliganded TR and TR/RXR reduce the local histone acetylation levels at the HIV-1 LTR while T-3 treatment reverses this reduction. We further demonstrated that unliganded TR recruits corepressors and at least one histone deacetylase. These results suggest that chromatin remodeling, including histone acetylation and chromatin disruption, is important for T-3 regulation of the HIV-1 LTR in vivo. C1 NICHHD, Unit Mol Morphogenesis, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA. RP Shi, YB (reprint author), NICHHD, Unit Mol Morphogenesis, Lab Gene Regulat & Dev, NIH, Bldg 18T,Rm 106, Bethesda, MD 20892 USA. OI Hsia, Victor/0000-0001-7623-2842 NR 73 TC 43 Z9 44 U1 0 U2 0 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 JUN PY 2002 VL 22 IS 12 BP 4043 EP 4052 DI 10.1128/MCB.22.12.4043-4052.2002 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 556UB UT WOS:000175866400009 PM 12024018 ER PT J AU Jang, IK Hu, RJ Lacana, E D'Adamio, L Gu, H AF Jang, IK Hu, RJ Lacana, E D'Adamio, L Gu, H TI Apoptosis-linked gene 2-deficient mice exhibit normal T-cell development and function SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID ANTI-FAS ANTIBODY; ALG-2 PROTEIN; CALPAIN; LYMPHOCYTES; MEMBER; GLUCOCORTICOIDS; RESISTANT; ADHESION; CLONING; DISEASE AB The apoptosis-linked gene product, ALG-2, is a member of the family of intracellular Cat(2+)-binding proteins and a part of the apoptotic machinery controlled by T-cell receptor (TCR), Fas, and glucocorticoid signals. To explore the physiologic function of ALG-2 in T-cell development and function, we generated mice harboring a null mutation in the alg-2 gene. The alg-2 null mutant mice were viable and fertile and showed neither gross developmental abnormality nor immune dysfunction. Analyses of apoptotic responses of ALG-2-deficient T cells demonstrated that ALG-2 deficiency failed to block apoptosis induced by TCR, Fas, or dexamethasone signals. These findings indicate that ALG-2 is physiologically dispensable for apoptotic responses induced by the above signaling pathways and suggest that other functionally redundant proteins might exist in mammalian cells. C1 NIAID, Immunol Lab, NIH, Rockville, MD 20852 USA. US FDA, Div Hematol Prod, Ctr Biol Evaluat & Res, Bethesda, MD 20892 USA. Yeshiva Univ Albert Einstein Coll Med, Dept Microbiol & Immunol, Bronx, NY 10461 USA. RP Gu, H (reprint author), NIAID, Immunol Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. OI D'Adamio, Luciano/0000-0002-9820-4882 NR 29 TC 38 Z9 38 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 JUN PY 2002 VL 22 IS 12 BP 4094 EP 4100 DI 10.1128/MCB.22.12.4094-4100.2002 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 556UB UT WOS:000175866400014 PM 12024023 ER PT J AU Tomasevic, N Peculis, BA AF Tomasevic, N Peculis, BA TI Xenopus LSm proteins bind U8 snoRNA via an internal evolutionarily conserved octamer sequence SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID SM-LIKE PROTEINS; SMALL NUCLEOLAR RNAS; RIBOSOMAL-RNA; MESSENGER-RNA; CRYSTAL-STRUCTURE; U6 SNRNA; SACCHAROMYCES-CEREVISIAE; PSEUDOURIDINE SYNTHASE; SPLICEOSOMAL SNRNPS; FUNCTIONAL-ANALYSIS AB U8 snoRNA plays a unique role in ribosome biogenesis: it is the only snoRNA essential for maturation of the large ribosomal subunit RNAs, 5.8S and 28S. To learn the mechanisms behind the in vivo role of U8 snoRNA, we have purified to near homogeneity and characterized a set of proteins responsible for the formation of a specific U8 RNA-binding complex. This 75-kDa complex is stable in the absence of added RNA and binds U8 with high specificity, requiring the conserved octamer sequence present in all U8 homologues. At least two proteins in this complex can be cross-linked directly to U8 RNA. We have identified the proteins as Xenopus homologues of the LSm (like Sm) proteins, which were previously reported to be involved in cytoplasmic degradation of mRNA and nuclear stabilization of U6 snRNA. We have identified LSm2, -3, -4, -6, -7, and -8 in our purified complex and found that this complex associates with U8 RNA in vivo. This purified complex can bind U6 snRNA in vitro but does not bind U3 or U14 snoRNA in vitro, demonstrating that the LSm complex specifically recognizes U8 RNA. C1 NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. RP Peculis, BA (reprint author), NIDDKD, Genet & Biochem Branch, NIH, 10 Ctr Dr,MSC 1766, Bethesda, MD 20892 USA. NR 60 TC 37 Z9 38 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 JUN PY 2002 VL 22 IS 12 BP 4101 EP 4112 DI 10.1128/MCB.22.12.410-4112.2002 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 556UB UT WOS:000175866400015 PM 12024024 ER PT J AU Deroo, BJ Rentsch, C Sampath, S Young, J DeFranco, DB Archer, TK AF Deroo, BJ Rentsch, C Sampath, S Young, J DeFranco, DB Archer, TK TI Proteasomal inhibition enhances glucocorticoid receptor transactivation and alters its subnuclear trafficking SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID TUMOR VIRUS PROMOTER; BREAST-CANCER CELLS; NUCLEAR MATRIX; IN-VIVO; DEPENDENT DEGRADATION; ESTROGEN-RECEPTOR; 26S PROTEASOME; TRANSCRIPTIONAL ACTIVATION; MEDIATED DEGRADATION; CHROMATIN STRUCTURE AB The ubiquitin-proteasome pathway regulates the turnover of many transcription factors, including steroid hormone receptors such as the estrogen receptor and progesterone receptor. For these receptors, proteasome inhibition interferes with steroid-mediated transcription. We show here that proteasome inhibition with MG132 results in increased accumulation of the glucocorticoid receptor (GR), confirming that it is likewise a substrate for the ubiquitin-proteasome degradative pathway. Using the mouse mammary tumor virus (MMTV) promoter integrated into tissue culture cells, we found that proteasome inhibition synergistically increases GR-mediated transactivation. This increased activation was observed in a number of cell lines and on various MMTV templates, either as transiently transfected reporters or stably integrated into chromatin. These observations suggest that the increase in GR-mediated transcription due to proteasome inhibition may occur downstream of the initial chromatin remodeling step. In support of this concept, the increase in transcription did not correlate with an increase in chromatin remodeling, as measured by restriction enzyme hypersensitivity, or transcription factor loading, as exemplified by nuclear factor 1. To investigate the relationship between GR turnover, transcription, and subnuclear trafficking, we examined the effect of proteasome inhibition on the mobility of the GR within the nucleus and association of the GR with the nuclear matrix. Blocking GR turnover reduced the mobility of the GR within the nucleus, and this correlated with increased association of the receptor with the nuclear matrix. As a result of proteasome inhibition, GR mobility within the nucleus was reduced while its association with the nuclear matrix was increased. Thus, while altered nuclear mobility of steroid receptors may be a common feature of proteasome inhibition, GR is unique in its enhanced transactivation activity that results when proteasome function is compromised. Proteasomes may therefore impact steroid receptor action at multiple levels and exert distinct effects on individual receptor types. C1 NIEHS, Chromatin & Gene Express Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. Univ Pittsburgh, Sch Med, Dept Pharmacol, Pittsburgh, PA 15261 USA. RP Archer, TK (reprint author), NIEHS, Chromatin & Gene Express Sect, Reprod & Dev Toxicol Lab, NIH, MD E4-06, Res Triangle Pk, NC 27709 USA. OI Rentsch, Claudia/0000-0002-3610-2020 NR 48 TC 108 Z9 112 U1 0 U2 0 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 JUN PY 2002 VL 22 IS 12 BP 4113 EP 4123 DI 10.1128/MCB.22.12.4113-4123.2002 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 556UB UT WOS:000175866400016 PM 12024025 ER PT J AU Safieddine, S Ly, CD Wang, YX Wang, CY Kachar, B Petralia, RS Wenthold, RJ AF Safieddine, S Ly, CD Wang, YX Wang, CY Kachar, B Petralia, RS Wenthold, RJ TI Ocsyn, a novel syntaxin-interacting protein enriched in the subapical region of inner hair cells SO MOLECULAR AND CELLULAR NEUROSCIENCE LA English DT Article ID UNCONVENTIONAL MYOSINS; ENDOSOMAL TRAFFICKING; ENDOPLASMIC-RETICULUM; MEMBRANE-PROTEINS; AFFERENT SYNAPSE; CUTICULAR PLATE; ENDOCYTOSIS; EXOCYTOSIS; DOMAINS; SNARES AB Sensory (hair) cells of the inner ear contain two specialized areas of membrane delivery. The first, located at the cell base, is the afferent synapse where rapid delivery of synaptic vesicles is required to convey information about auditory signals with exceedingly high temporal precision. The second area is at the apex. To accommodate the continuous movement of stereocilia and facilitate their repair, recycling of membrane components is required. Intense vesicular traffic is restricted to a narrow band of cytoplasm around the cuticular plate, which anchors stereocilia. Our previous analyses showed that SNARE proteins (syntaxin 1A/SNAP25/VAMP1) are concentrated at both poles of hair cells, consistent with their involvement in membrane delivery at both locations. To investigate further the molecules involved in membrane delivery at these two sites, we constructed a two-hybrid library of the organ of Corti and probed it with syntaxin 1A. Here we report the cloning of a novel syntaxin-binding protein that is concentrated in a previously uncharacterized organelle at the apex of inner hair cells. C1 Natl Inst Deafness & Other Commun Disorders, NIH, Lab Neurochem, Bethesda, MD 20892 USA. Natl Inst Deafness & Other Commun Disorders, NIH, Lab Cell Biol, Bethesda, MD 20892 USA. RP Wenthold, RJ (reprint author), Natl Inst Deafness & Other Commun Disorders, NIH, Lab Neurochem, Bethesda, MD 20892 USA. NR 52 TC 11 Z9 11 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 1044-7431 J9 MOL CELL NEUROSCI JI Mol. Cell. Neurosci. PD JUN PY 2002 VL 20 IS 2 BP 343 EP 353 DI 10.1006/mcne.2002.1120 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 569YQ UT WOS:000176631200014 PM 12093165 ER PT J AU Wang, YJ Zhou, YX Szabo, K Haft, CR Trejo, J AF Wang, YJ Zhou, YX Szabo, K Haft, CR Trejo, J TI Down-regulation of protease-activated receptor-1 is regulated by sorting nexin 1 SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID THROMBIN-RECEPTOR; BETA(2)-ADRENERGIC RECEPTOR; INTRACELLULAR TRAFFICKING; ENDOTHELIAL-CELLS; MEMBRANE-PROTEINS; COUPLED RECEPTOR; YEAST; MECHANISM; GOLGI; RESENSITIZATION AB Degradation or "down-regulation" of protease-activated receptor-1 (PAR1), a G protein-coupled receptor for thrombin, is critical for termination of receptor signaling. Toward understanding the molecular mechanisms by which activated PAR1 is internalized, sorted to lysosomes, and degraded, we investigated whether PARI interacted with sorting nexin 1 (SNX1). SNX1 is a membrane-associated protein that functions in lysosomal sorting of the epidermal growth factor receptor. In vitro biochemical binding assays revealed a specific interaction between a glutathione S-transferase fusion of SNX1 and PAR1. In HeLa cells, activated PAR1 colocalized with endogenous SNX1 and coimmunoprecipitated SNX1. SNX1 contains a phox homoloo- domain predicted to bind phosphatidylinositol-3-phosphate and a C-terminal coiled-coil region. To assess SNX1 function, we examined the effects of SNX1 deletion mutants on PAR1 trafficking. Neither the N terminus nor phox homology domain of SNX1 affected PARI trafficking. By contrast, overexpression of SNX1 C-terminal domain markedly inhibited agonist-induced degradation of PARI, whereas internalization remained virtually intact. Immunofluorescence microscopy studies revealed substantial PARI accumulation in an early endosome antigen-1-positive compartment in agonist-treated cells expressing SNX1 C terminus. By contrast, lysosome-associated membrane protein-1 distribution was unperturbed. Together, these findings strongly suggest a role for SNX1 in sorting of PAR1 from early endosomes to lysosomes. Moreover, this study provides the first example of a protein involved in lysosomal sorting of a G protein-coupled receptor in mammalian cells. C1 Univ N Carolina, Sch Med, Dept Pharmacol, Chapel Hill, NC 27599 USA. Univ N Carolina, Sch Med, Dept Cell & Mol Physiol, Chapel Hill, NC 27599 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. RP Trejo, J (reprint author), Univ N Carolina, Sch Med, Dept Pharmacol, Chapel Hill, NC 27599 USA. FU NHLBI NIH HHS [HL67697, K01 HL067697] NR 38 TC 94 Z9 99 U1 0 U2 2 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD JUN PY 2002 VL 13 IS 6 BP 1965 EP 1976 DI 10.1091/mbc.E01-11-0131 PG 12 WC Cell Biology SC Cell Biology GA 566GP UT WOS:000176418800015 PM 12058063 ER PT J AU Elbi, C Misteli, T Hager, GL AF Elbi, C Misteli, T Hager, GL TI Recruitment of dioxin receptor to active transcription sites SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID ARYL-HYDROCARBON RECEPTOR; NUCLEAR TRANSLOCATOR PROTEIN; RNA-POLYMERASE-II; DROSOPHILA PERIOD GENE; AH-RECEPTOR; MICE LACKING; MINERALOCORTICOID RECEPTORS; ACTIVATION FUNCTION; SPLICING FACTORS; LIVING CELLS AB The aryl hydrocarbon receptor (AhR or dioxin receptor) is a ligand-activated transcription factor that heterodimerizes with the AhR nuclear translocator (ARNT/HIF-1beta) to form an AhR/ARNT transcription factor complex. This complex binds to specific DNA sites in the regulatory domains of numerous target genes and mediates the biological effects of exogenous ligands. Herein, we have investigated the subcellular distribution of the AhR/ARNT complex in response to ligand stimulation, by using live-cell confocal and high-resolution deconvolution microscopy. We found that unliganded AhR shows a predominantly cytoplasmic diffuse distribution in mouse hepatoma cells. On addition of ligand, AhR rapidly translocates to the nucleus and accumulates in multiple bright foci. Inhibition of transcription prevented the formation of AhR foci. Dual- and triple-immunolabeling experiments, combined with labeling of nascent RNA, showed that the foci are transcription sites, indicating that upon ligand stimulation, AhR is recruited to active transcription sites. The interaction of AhR with ARNT was both necessary and sufficient for the recruitment of AhR to transcription sites. These results indicate that AhR/ARNT complexes are recruited to specific subnuclear compartments in a ligand-dependent manner and that these foci represent the sites of AhR target genes. C1 NCI, Lab Receptor Biol & Gene Express, NIH, Bethesda, MD 20892 USA. RP Hager, GL (reprint author), NCI, Lab Receptor Biol & Gene Express, NIH, Bethesda, MD 20892 USA. NR 54 TC 55 Z9 56 U1 1 U2 3 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD JUN PY 2002 VL 13 IS 6 BP 2001 EP 2015 DI 10.1091/mbc.02-01-0009 PG 15 WC Cell Biology SC Cell Biology GA 566GP UT WOS:000176418800017 PM 12058065 ER PT J AU Noy, R Ben-Zvi, Z Manor, E Candotti, F Morris, JC Ford, H Marquez, VE Johns, DG Agbaria, R AF Noy, R Ben-Zvi, Z Manor, E Candotti, F Morris, JC Ford, H Marquez, VE Johns, DG Agbaria, R TI Antitumor activity and metabolic activation of N-methanocarbathymidine, a novel thymidine analogue with a pseudosugar rigidly fixed in the northern conformation, in murine colon cancer cells expressing herpes simplex thymidine kinase SO MOLECULAR CANCER THERAPEUTICS LA English DT Article ID PURINE NUCLEOSIDE ANALOGS; VIRUS TYPE-1; GENE-THERAPY; BRAIN-TUMORS; (E)-5-(2-BROMOVINYL)-2'-DEOXYURIDINE; 9-(1,3-DIHYDROXY-2-PROPOXYMETHYL)GUANINE; MECHANISM; LEUKEMIA; SUPERIOR; PARADIGM AB N-Methanocarbathymidine [(N)-MCT], a thymidine analogue incorporating a pseudosugar with a fixed Northern conformation, exhibits antiherpetic activity against both herpes simplex virus (HSV) HSV-1 and HSV-2, with a potency greater than that of the reference standard, ganciclovir (GCV). In the present study, we have assessed the cytotoxic activity in vitro of (N)-MCT in wild-type murine colon cancer cells (MC38) and in cells expressing the herpes simplex thymidine kinase gene (MC38/HSV-tk), and the antitumor activity of (N)-MCT in vivo against HSV-tk transduced and nontransduced MC38 murine tumors. In vitro, when assessed over a 48-h period, the growth-inhibitory activity (IC50) of (N)-MCT toward MC38/HSV-tk cells was 2.9 muM. In parallel studies, the cytostatic activity of the reference compound GCV in these tumor lines was 3.0 muM. In studies in vivo, both (N)-MCT and GCV (100 mg/kg) given twice daily for 7 days completely inhibited the growth of HSV-tk-transduced MC38 tumors while exhibiting no effect on nontransduced MC38 tumors in mice. In nontransduced cells both in vitro and in vivo, only low levels of (N)-MCT and its monophosphate could be detected after administration of the parent drug, whereas in HSV-tk-transduced cells (N)-MCT was phosphorylated to its respective mono-, di-, and triphosphates. Furthermore, data showed that (N)-MCT incorporated in high levels into cellular DNA whereas trace levels were measured into RNA. These observations indicate that (N)-MCT may be a useful candidate prodrug for HSV-tk suicide gene therapy of cancer. C1 Ben Gurion Univ Negev, Fac Hlth Sci, Dept Clin Pharmacol, IL-84105 Beer Sheva, Israel. NHGRI, NCI, NIH, Frederick, MD 21702 USA. NCI, Med Chem Lab, NIH, Frederick, MD 21702 USA. NCI, Metab Branch, Ctr Canc Res, Bethesda, MD 20892 USA. RP Agbaria, R (reprint author), Ben Gurion Univ Negev, Fac Hlth Sci, Dept Clin Pharmacol, IL-84105 Beer Sheva, Israel. NR 33 TC 15 Z9 15 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1535-7163 J9 MOL CANCER THER JI Mol. Cancer Ther. PD JUN PY 2002 VL 1 IS 8 BP 585 EP 593 PG 9 WC Oncology SC Oncology GA 607BL UT WOS:000178770500003 PM 12479218 ER PT J AU Fan, D Yano, S Shinohara, H Solorzano, C Van Arsdall, M Bucana, CD Pathak, S Kruzel, E Herbst, RS Onn, A Roach, JS Onda, M Wang, QC Pastan, I Fidler, IJ AF Fan, D Yano, S Shinohara, H Solorzano, C Van Arsdall, M Bucana, CD Pathak, S Kruzel, E Herbst, RS Onn, A Roach, JS Onda, M Wang, QC Pastan, I Fidler, IJ TI Targeted therapy against human lung cancer in nude mice by high-affinity recombinant antimesothelin single-chain Fv immunotoxin SO MOLECULAR CANCER THERAPEUTICS LA English DT Article ID PSEUDOMONAS EXOTOXIN; ANTITUMOR-ACTIVITY; MOLECULAR-CLONING; TYROSINE KINASE; OVARIAN CANCERS; PHAGE DISPLAY; CELL LYMPHOMA; ANTIBODY; MESOTHELIN; EXPRESSION AB Several tumors, including mesothelioma and ovarian cancer, can overexpress mesothelin, a glycosylphosphatidylinositol-linked differentiation glycoprotein. The membrane-bound type of mesothelin is found in the blood of cancer patients at a very low level, which makes mesothelin a good candidate for targeted therapy of certain cancers. An antimesothelin disulfide-linked Fv (SS1 Fv) was fused to a truncated mutant of Pseudomonas exotoxin A to produce the recombinant immunotoxin SS1(dsFv)-PE38, which has a high binding affinity to mesothelin (K-d = 0.7 nM). Our studies in vitro showed that SS1(dsFv)-PE38 is significantly more cytotoxic to the high-mesothelin-producing NCI-H226 human non-small cell lung cancer cells than to human lung adenocarcinoma PC14PE6 cells, which do not express mesothelin. When administered at a nontoxic dose of 500 mug/kg on days 7, 9, and 11 to nude mice injected i.v. with the two human lung cancer cell lines, SS1(dsFv)-PE38 selectively inhibited experimental lung metastases produced by the mesothelin-producing NCI-H226 cells. Our data indicate that mesothelin-producing squamous cell carcinoma of the lung may be a good target for this immunotoxin. C1 Univ Texas, MD Anderson Canc Ctr, Dept Canc Biol, Houston, TX 77030 USA. Univ Texas, MD Anderson Canc Ctr, Dept Thorac Head & Neck Med Oncol, Houston, TX 77030 USA. NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Fan, D (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Canc Biol 173, 1515 Holcombe Blvd, Houston, TX 77030 USA. FU NCI NIH HHS [CA16672, R35-CA42107] NR 34 TC 39 Z9 41 U1 2 U2 4 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1535-7163 J9 MOL CANCER THER JI Mol. Cancer Ther. PD JUN PY 2002 VL 1 IS 8 BP 595 EP 600 PG 6 WC Oncology SC Oncology GA 607BL UT WOS:000178770500004 PM 12479219 ER PT J AU Young, MR Farrell, L Lambert, P Awasthi, P Colburn, NH AF Young, MR Farrell, L Lambert, P Awasthi, P Colburn, NH TI Protection against human papillomavirus type 16-E7 oncogene-induced tumorigenesis by in vivo expression of dominant-negative c-jun SO MOLECULAR CARCINOGENESIS LA English DT Article DE activator protein-1; human papillomavirus type 16; tumor promotion; TAM67; cancer prevention ID SKIN TUMOR-DEVELOPMENT; NF-KAPPA-B; TRANSGENIC MICE; IN-VIVO; HUMAN KERATINOCYTES; CELL-PROLIFERATION; E7 ONCOGENE; CARCINOGENESIS; AP-1; TRANSACTIVATION AB Expression of the human papillomavirus (HPV) type 16 E6 and E7 gene products is a risk factor for human cervical carcinogenesis as well as skin and oral carcinogenesis. Expression of the HPV-16 E7 gene in mouse skin induces hyperplasia and enhances tumor promotion. Expression of dominant-negative c-jun (TAM67) in the mouse skin protects mice from 7,12-dimethylbenz[a]anthracene (DMBA)/12-O-tetradecanoylphorbol-13-acetate (TPA)-induced papillomagenesis without blocking mitogen-induced hyperproliferation. To determine the role of activator protein-1 (AP-1) in HPV-induced cancer, we crossed HPV-16 E7 mice with TAM67 mice and analyzed the effects of DMBA/TPA on tumor promotion. We showed that expression of TAM67 protected mice from HPV-16 E7-enhanced tumorigenesis, suggesting AP-1 as a target for prevention of HPV-induced cancer. Published 2002 Wiley-Liss, Inc.(dagger) C1 NCI, Basic Res Lab, Frederick, MD 21702 USA. Univ Wisconsin, Sch Med, Dept Pathol, McArdle Lab Canc Res, Madison, WI 53706 USA. SAIC Frederick, Lab Anim Sci Program, Frederick, MD USA. RP Young, MR (reprint author), NCI, Basic Res Lab, POB B, Frederick, MD 21702 USA. NR 22 TC 29 Z9 30 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0899-1987 J9 MOL CARCINOGEN JI Mol. Carcinog. PD JUN PY 2002 VL 34 IS 2 BP 72 EP 77 DI 10.1002/mc.10050 PG 6 WC Biochemistry & Molecular Biology; Oncology SC Biochemistry & Molecular Biology; Oncology GA 559NK UT WOS:000176032100003 PM 12112313 ER PT J AU Lobenhofer, EK Bennett, L Cable, PL Li, LP Bushel, PR Afshari, CA AF Lobenhofer, EK Bennett, L Cable, PL Li, LP Bushel, PR Afshari, CA TI Regulation of DNA replication fork genes by 17 beta-estradiol SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID BREAST-CANCER-CELLS; ACTIVATED PROTEIN-KINASE; MYC ONCOGENE EXPRESSION; ESTROGEN-RECEPTOR-BETA; MCF-7 CELLS; TRANSCRIPTIONAL ACTIVATION; PROGESTERONE-RECEPTOR; CYCLIN E-CDK2; RESPONSE ELEMENT; TISSUE-CULTURE AB The steroid hormone estrogen can stimulate mitogenesis in hormone-responsive breast cancer epithelial cells. This action is attributed to the transcriptional activity of the ER, a ligand-dependent transcription factor. However, the exact molecular mechanism underlying estrogen-induced proliferation has yet to be completely elucidated. Using custom cDNA microarrays containing many genes implicated in cell cycle progression and DNA replication, we examined the gene expression of a hormone-responsive breast cancer cell line (MCF-7) treated with a mitogenic dose of estrogen in the absence of confounding growth factors found in serum. Gene expression changes were monitored 1, 4, 12, 24, 36, and 48 h after estrogen stimulation so that RNA levels at critical times throughout cell cycle progression could be monitored. Significant changes include the altered transcript levels of genes implicated in transcription, cellular signaling, and cell cycle checkpoints. At time points during which increased numbers of cells were progressing through S phase, a majority of the genes associated with the DNA replication fork were also found to be induced. The coexpression of DNA replication fork genes by estrogen without the support of serum growth factors indicates an important estrogen regulatory component of the molecular mechanism driving estrogen-induced mitogenesis. C1 NIEHS, Gene Regulat Grp, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Carcinogenesis Lab, Microarray Ctr, Res Triangle Pk, NC 27709 USA. NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. RP Lobenhofer, EK (reprint author), NIEHS, Gene Regulat Grp, POB 12233 MD2-04, Res Triangle Pk, NC 27709 USA. NR 64 TC 61 Z9 64 U1 0 U2 0 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD JUN PY 2002 VL 16 IS 6 BP 1215 EP 1229 DI 10.1210/me.16.6.1215 PG 15 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 557FK UT WOS:000175897500009 PM 12040010 ER PT J AU Shillingford, JM Miyoshi, K Flagella, M Shull, GE Hennighausen, L AF Shillingford, JM Miyoshi, K Flagella, M Shull, GE Hennighausen, L TI Mouse mammary epithelial cells express the Na-K-Cl cotransporter, NKCC1: Characterization, localization, and involvement in ductal development and morphogenesis SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID NA+-K+-2CL(-) COTRANSPORTER; GLUCOSE TRANSPORTERS; GLAND DEVELOPMENT; PROLIFERATION; CHLORIDE; VOLUME; MICE; GENE; FIBROBLASTS; MECHANISMS AB Despite the fact that physiological evidence points to the existence of a functional Na-K-Cl cotransporter in the mammary gland, the molecular identity of this transport process remains unknown. We now show that the Na-K-Cl cotransporter isoform, NKCC1, is expressed in mammary tissue. Developmental profiling revealed that the level of NKCC1 protein was significantly influenced by the stage of mammary gland development, and immunolocalization studies demonstrated that NKCC1 was present on the basolateral membrane of mammary epithelial cells. To examine whether functional NKCC1 is required for mammary epithelial cell development, we used NKCC1 -/- mice. We demonstrate that NKCC1 -/- mammary epithelium exhibited a significant delay in ductal outgrowth and an increase in branching morphogenesis during virgin development. These effects were autonomous to the epithelium as assessed by mammary gland transplantation. Although the absence of NKCC1 had no apparent effect on gross mammary epithelial cell morphology during lactation, pups born to NKCC1 -/- mice failed to thrive. Finally, analysis of NKCC1 protein in mouse models that exhibit defects in mammary gland development demonstrate that high levels of NKCC1 protein are indicative of ductal epithelial cells, and the presence of NKCC1 protein is characteristic of mammary epithelial cell identity. C1 NIH, Lab Genet & Physiol, Bethesda, MD 20892 USA. Univ Tokushima, Sch Dent, Dept Biochem, Tokushima 7708504, Japan. Univ Cincinnati, Dept Mol Genet Biochem & Microbiol, Cincinnati, OH 45221 USA. RP Shillingford, JM (reprint author), NIH, Lab Genet & Physiol, Bldg 8,Room 105,9000 Rockville Pike, Bethesda, MD 20892 USA. FU NIDDK NIH HHS [DK-50594] NR 40 TC 35 Z9 35 U1 1 U2 3 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD JUN PY 2002 VL 16 IS 6 BP 1309 EP 1321 DI 10.1210/me.16.6.1309 PG 13 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 557FK UT WOS:000175897500016 PM 12040017 ER PT J AU Rocher, C Letellier, T Copeland, C Lestienne, P AF Rocher, C Letellier, T Copeland, C Lestienne, P TI Base composition at mtDNA boundaries suggests a DNA triple helix model for human mitochondrial DNA large-scale rearrangements SO MOLECULAR GENETICS AND METABOLISM LA English DT Article DE slipped mispairing; deletion; duplication; mitochondrial DNA ID TRANSFER-RNA SYNTHETASES; P55 ACCESSORY SUBUNIT; KEARNS-SAYRE SYNDROME; POLYMERASE-GAMMA; CATALYTIC SUBUNIT; CRYSTAL-STRUCTURE; HUMAN-CELLS; DELETIONS; RECOMBINATION; REPLICATION AB Different mechanisms have been proposed to account for mitochondrial DNA (mtDNA) instability based on the presence of short homologous sequences (direct repeats, DR) at the potential boundaries of mtDNA rearrangements. Among them, slippage-mispairing of the replication complex during the asymmetric replication cycle of the mammalian mitochondrial DNA has been proposed to account for the preferential localization of deletions. This mechanism involves a transfer of the replication complex from the first neo-synthesized heavy (H) strand of the DR1, to the DR2, thus bypassing the intervening sequence and producing a deleted molecule. Nevertheless. the nature of the bonds between the DNA strands remains unknown as the forward sequence of DR2, beyond the replication complex, stays double-stranded. Here. we have analyzed the base composition of the DR at the boundaries of mtDNA deletions and duplications and found a skewed pyrimidine content of about 75% in the light-strand DNA template. This suggests the possible building of a DNA triple helix between the G-rich neo-synthesized DR1 and the base-paired homologous G.C-rich DR-2. In vitro experiments with the purified human DNA polymerase gamma subunits enabled us to show that the third DNA strand may be used as a primer for DNA replication, using a template with the direct repeat forming a hairpin, with which the primer could initiate DNA replication. These data suggest a novel molecular basis for mitochondrial DNA rearrangements through the distributive nature of the DNA polymerase gamma, at the level of the direct repeats. A general model accounting for large-scale mitochondrial DNA deletion and duplication is proposed. These experiments extend to a DNA polymerase from an eucaryote source the use of a DNA triple helix strand as a primer, like other DNA polymerases from phage and bacterial origins. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Univ Victor Segalen Bordeaux 2, INSERM, EMI 99 29, F-33076 Bordeaux, France. NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Lestienne, P (reprint author), Univ Victor Segalen Bordeaux 2, INSERM, EMI 99 29, 146 Rue Leo Saignat, F-33076 Bordeaux, France. NR 49 TC 21 Z9 21 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 1096-7192 J9 MOL GENET METAB JI Mol. Genet. Metab. PD JUN PY 2002 VL 76 IS 2 BP 123 EP 132 AR PII S1096-7192(02)00024-0 DI 10.1016/S1096-7192(02)00024-0 PG 10 WC Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental SC Endocrinology & Metabolism; Genetics & Heredity; Research & Experimental Medicine GA 571YW UT WOS:000176747600005 PM 12083809 ER PT J AU Girschick, HJ Lipsky, PE AF Girschick, HJ Lipsky, PE TI The kappa gene repertoire of human neonatal B cells SO MOLECULAR IMMUNOLOGY LA English DT Article DE cord blood; CD5; B lymphocytes; V kappa repertoire; secondary gene rearrangement ID JUVENILE RHEUMATOID-ARTHRITIS; SYSTEMIC LUPUS-ERYTHEMATOSUS; BLOOD ANTIBODY REPERTOIRE; VARIABLE REGION GENES; J-LAMBDA REPERTOIRE; SOMATIC HYPERMUTATION; IMMUNOGLOBULIN GENES; PREFERENTIAL UTILIZATION; MOLECULAR MECHANISMS; V(D)J RECOMBINATION AB The kappa chain repertoire of individual IgD(+) human neonatal B cells was analyzed using a single cell PCR technique. A total of 104 productive and 90 non-productive VkappaJkappa rearrangements from three cord blood B cell samples were sequenced and compared to the adult IgM(+) peripheral B cell VkappaJkappa repertoire. All six Vkappa families were present in neonatal B cells, but the distribution was not random, In the non-productive repertoire Vkappa2 and Vkappa6 families were less frequent, Vkappa1 and Vkappa3 families were as frequent, and Vkappa4 and Vkappa5 families were more frequent than expected from random chance. Notably, the Vkappa2 family was negatively selected into the productive repertoire. In contrast. the Vkappa1 family was positively selected because of positive selection of three specific genes, O12/O2, L12a and L9. B3 (Vkappa4) and B2 (Vkappa5) were over-represented in the non-productive repertoire and then were expressed less frequently in the productive repertoire, In contrast. the Vkappa3 family gene. A27, was also over-represented in the non-productive repertoire but not further selected into the productive repertoire. Compared to the adult repertoire, junctional diversity was less marked because of a diminished influence of TdT activity, whereas the mean CDR3 length was comparable to that of normal adult B cells. Comparison of the distribution of Vkappa and Jkappa genes with those found in normal adult subjects suggested that there was less receptor editing in neonatal B cells, When neonatal CD5(+) B cells were compared with CD5(-) IgD(+) B cells, it was noted that the Vkappa gene A30 was used only in CD5(+) B cells in both the productive and non-productive repertoires. The results indicate that the Usage of Vkappa genes by neonatal B cells is biased by both intrinsic molecular processes and selection. The evidence of selection indicates that the Vkappa repertoire is shaped by self antigens, since exposure to exogenous antigens is limited at the time of birth. (C) 2002 Published by Elsevier Science Ltd. C1 Univ Texas, SW Med Ctr, Harold C Simmons Arthritis Res Ctr, Dept Internal Med, Dallas, TX 75235 USA. Univ Wurzburg, Childrens Hosp, D-97070 Wurzburg, Germany. RP Girschick, HJ (reprint author), NIAMS, NIH, 9000 Rockville Pike,Room 9N228, Bethesda, MD 20892 USA. FU NIAID NIH HHS [AI 31229] NR 48 TC 9 Z9 9 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 JUN PY 2002 VL 38 IS 15 BP 1113 EP 1127 AR PII S0161-5890(02)00039-1 DI 10.1016/S0161-5890(02)00039-1 PG 15 WC Biochemistry & Molecular Biology; Immunology SC Biochemistry & Molecular Biology; Immunology GA 573UV UT WOS:000176850800003 PM 12044778 ER PT J AU Roh, DH Bowers, B Riezman, H Cabib, E AF Roh, DH Bowers, B Riezman, H Cabib, E TI Rho1p mutations specific for regulation of beta(1 -> 3)glucan synthesis and the order of assembly of the yeast cell wall SO MOLECULAR MICROBIOLOGY LA English DT Article ID GTP-BINDING PROTEIN; SACCHAROMYCES-CEREVISIAE; CRYSTAL-STRUCTURE; ALPHA-AGGLUTININ; CROSS-LINKING; KINASE-C; MUTANTS; CHITIN; BETA-1,6-GLUCAN; IDENTIFICATION AB In the yeast Saccharomyces cerevisiae , the GTP-binding protein Rho1 is required for beta(1-->3)glucan synthase activity, for activation of protein kinase C and the cell integrity pathway and for progression in G1, cell polarization and exocytosis. A genetic screen for cells that become permeabilized at non-permissive temperature was used to isolate in vitro -generated mutants of Rho1p. After undergoing a battery of tests, several of them appeared to be specifically defective in the beta(1-->3)glucan synthesis function of Rho1p. At the non-permissive temperature (37degreesC), the mutants developed defects in the cell wall, especially at the tip of new buds. In the yeast cell wall, beta(1-->6)glucan is linked to both beta(1-->3)glucan and mannoprotein, as well as occasionally to chitin. We have used the rho1 mutants to study the order of assembly of the cell wall components. The incorporation of [(14) C]-glucose into beta(1-->3)glucan at 37degreesC was decreased or abolished in the mutants. Concomitantly, a partial defect in the incorporation of label into cell wall mannoproteins and beta(1-->6)glucan was observed. In contrast, YW3458, an inhibitor of glycosylphosphatidylinositol anchor formation, prevented mannoprotein incorporation, whereas the beta(1-->3)-beta(1-->6)glucan complex was synthesized at almost normal levels. As beta(1-->3)glucan can be synthesized in vitro or in vivo independently, we conclude that the order of addition in vivo is beta(1-->3)glucan, beta(1-->6)glucan, mannoprotein. Previous observations indicate that chitin is the last component to be incorporated into the complex. C1 NIDDKD, Lab Biochem & Genet, Bethesda, MD 20892 USA. NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Basel, Biozentrum, CH-4056 Basel, Switzerland. RP Cabib, E (reprint author), NIDDKD, Lab Biochem & Genet, Bldg 8,Room 403, Bethesda, MD 20892 USA. NR 54 TC 26 Z9 30 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD JUN PY 2002 VL 44 IS 5 BP 1167 EP 1183 DI 10.1046/j.1365-2958.2002.02955.x PG 17 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 556GQ UT WOS:000175841400005 PM 12068804 ER PT J AU Martin, RG Rosner, JL AF Martin, RG Rosner, JL TI Genomics of the marA/soxS/rob regulon of Escherichia coli: identification of directly activated promoters by application of molecular genetics and informatics to microarray data SO MOLECULAR MICROBIOLOGY LA English DT Article ID MULTIPLE ANTIBIOTIC-RESISTANCE; TRANSCRIPTIONAL ACTIVATION; DNA-BINDING; SUPEROXIDE STRESS; SOXS; MARA; GENES; EXPRESSION; SEQUENCE; ROB AB Microarray analyses are providing a plethora of data concerning transcriptional responses to specific gene regulators and their inducers but do not distinguish between direct and indirect responses. Here, we identify directly activated promoters of the overlapping marA , soxS and rob regulon(s) of Escherichia coli by applying informatics, genomics and molecular genetics to microarray data obtained by others. Those studies found that overexpression of marA , or the treatment of cells with salicylate to derepress marA , or treatment with paraquat to induce soxS , resulted in elevated transcription of 153 genes. However, only 27 out of the promoters showed increased transcription under at least two of the aforementioned conditions and eight of those were previously known to be directly activated. A computer algorithm was used to identify potential activator binding sites located upstream of the remaining 19 promoters of this subset, and conventional genetic and biochemical approaches were applied to test whether these sites are critical for activation by the homologous MarA, SoxS and Rob transcriptional activators. Only seven out of the 19 promoters were found to be activated when fused to lacZ and tested as single lysogens. All seven contained an essential activator binding site. The remaining promoters were insensitive to stimulation by the inducers suggesting that the great majority of elevated microarray transcripts either were misidentified or resulted from indirect effects requiring sequences outside of the promoter region. We estimate that the total number of directly activated promoters in the regulon is less than 40. C1 NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Martin, RG (reprint author), NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NR 35 TC 89 Z9 95 U1 0 U2 2 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD JUN PY 2002 VL 44 IS 6 BP 1611 EP 1624 DI 10.1046/j.1365-2958.2002.02985.x PG 14 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 562UB UT WOS:000176216400016 PM 12067348 ER PT J AU Li, MD Konu, O Kane, JK Becker, KG AF Li, MD Konu, O Kane, JK Becker, KG TI Microarray technology and its application on nicotine research SO MOLECULAR NEUROBIOLOGY LA English DT Review DE nicotine; drug abuse; microarray; expression; pathways; review ID GROWTH-FACTOR-BETA; POLYMERASE CHAIN-REACTION; ADRENAL CHROMAFFIN CELLS; FACTOR-KAPPA-B; GENE-EXPRESSION; RAT-BRAIN; ACETYLCHOLINE-RELEASE; CIGARETTE-SMOKING; CDNA MICROARRAYS; CORTICAL-NEURONS AB Since its development, microarray technique has revolutionized almost all fields of biomedical research by enabling high-throughput gene expression profiling. Using cDNA microarrays, thousands of genes from various organisms have been examined with respect to differentiation/development, disease diagnosis, and drug discovery. Nevertheless, research on nicotine using cDNA microarrays has been rather limited. Therefore, it is our intention in this article to report the findings of our cDNA microarray study on nicotine. We first present an overview of the microarray technology, particularly focusing on the factors related to microarray design and analysis. Second, we provide a detailed description of several newly identified biological pathways in our laboratory, such as phosphatidylinositol signaling and calcium homeostasis, which are involved in response to chronic nicotine administration. Additionally, we illustrate how comparisons between microarray studies help identify candidate genes that potentially may explain the observed inverse association between smoking and schizophrenia. Lastly, given the early stage of microarray research on nicotine, we elaborate on the need for an efficient analysis of genetic networks to further enhance our understanding of the mechanisms involved in nicotine abuse and addiction. C1 Univ Tennessee, Coll Med, Dept Pharmacol, Memphis, TN 38163 USA. NIA, NIH, Baltimore, MD 21224 USA. RP Li, MD (reprint author), 874 Union Ave,Suite 115, Memphis, TN 38163 USA. OI Becker, Kevin/0000-0002-6794-6656 FU NIDA NIH HHS [DA-12844, R01-DA13783] NR 101 TC 35 Z9 35 U1 0 U2 1 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0893-7648 J9 MOL NEUROBIOL JI Mol. Neurobiol. PD JUN PY 2002 VL 25 IS 3 BP 265 EP 285 DI 10.1385/MN:25:3:265 PG 21 WC Neurosciences SC Neurosciences & Neurology GA 565LD UT WOS:000176371000004 PM 12109875 ER PT J AU Ueda, A Kakizaki, S Negishi, M Sueyoshi, T AF Ueda, A Kakizaki, S Negishi, M Sueyoshi, T TI Residue threonine 350 confers steroid hormone responsiveness to the mouse nuclear orphan receptor CAR SO MOLECULAR PHARMACOLOGY LA English DT Article ID RAT CYP2B2 GENE; CYTOCHROME-P450 GENES; XENOBIOTIC INDUCTION; 5'-FLANKING REGION; ENHANCER MODULE; TRANSLOCATION; ACTIVATION; LIVER AB Steroid hormones modulate activity of the nuclear receptor constitutive active receptor (CAR, or constitutive androstane receptor) in mouse liver. Progesterone and testosterone repress the constitutive activity of mouse CAR (mCAR) in cell-mediated transfection assays, whereas estrogens activate the repressed receptor. This repression and activation is not observed with human CAR. To define the structural basis that confers the hormone responsiveness to mCAR, we constructed various chimeric and mutated receptors and examined their response to steroid hormones. The hormone responsiveness resided near or within AF-2 domain of mCAR. Moreover, a single mutation of threonine at position 350 to the corresponding methionine in the human counterpart abolished the repression of mCAR by steroid hormones. Coactivation by steroid receptor coactivator 1 (SRC-1) of mCAR did not depend on the threonine 350. However, overexpression of SRC-1 counteracted progesterone to repress mCAR activity. Thus, threonine 350 seems to regulate hormone responsiveness of mCAR by interfering indirectly an interaction of the receptor with a coactivator. C1 NIEHS, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Negishi, M (reprint author), NIEHS, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 20 TC 22 Z9 23 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD JUN PY 2002 VL 61 IS 6 BP 1284 EP 1288 DI 10.1124/mol.61.6.1284 PG 5 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LH UT WOS:000175736600003 PM 12021388 ER PT J AU Tomic, M Van Goor, F He, ML Zivadinovic, D Stojilkovic, SS AF Tomic, M Van Goor, F He, ML Zivadinovic, D Stojilkovic, SS TI Ca2+-mobilizing endothelin-A receptors inhibit voltage- gated Ca2+ influx through G(i/o) signaling pathway in pituitary lactotrophs SO MOLECULAR PHARMACOLOGY LA English DT Article ID THYROTROPIN-RELEASING-HORMONE; LONG-TERM EXPOSURE; RAT LACTOTROPHS; CELL-LINE; POTASSIUM CURRENT; CALCIUM; SECRETION; CHANNELS; SOMATOSTATIN; EXPRESSION AB In excitable cells, receptor-induced Ca2+ release from intracellular stores is usually accompanied by sustained depolarization of cells and facilitated voltage-gated Ca2+ influx (VGCI). In quiescent pituitary lactotrophs, however, endothelin-1 (ET-1) induced rapid Ca2+ release without triggering Ca2+ influx. Furthermore, in spontaneously firing and depolarized lactotrophs, the Ca2+-mobilizing action of ET-1 was followed by inhibition of spontaneous VGCI caused by prolonged cell hyperpolarization and abolition of action potential-driven Ca2+ influx. Agonist-induced depolarization of cells and enhancement of VGCI upon Ca2+ mobilization was established in both quiescent and firing lactotrophs treated overnight with pertussis toxin (PTX). Activation of adenylyl cyclase by forskolin and addition of cell-permeable 8-bromo-cAMP did not affect ET-1-induced sustained inhibition of VGCI, suggesting that the cAMP-protein kinase A signaling pathway does not mediate the inhibitory action of ET-1 on VGCI. Consistent with the role of PTX-sensitive K+ channels in ET-1-induced hyperpolarization of control cells, but not PTX-treated cells, ET-1 decreased the cell input resistance and activated a 5 mM Cs+-sensitive K+ current. In the presence of Cs+, ET-1 stimulated VGCI in a manner comparable with that observed in PTX-treated cells, whereas E-4031, a specific blocker of ether-a-go-go-related gene-like K+ channels, was ineffective. Similar effects of PTX and Cs+ were also observed in GH(3) immortalized cells transiently expressing ETA receptors. These results indicate that signaling of ETA receptors through the G(i/o) pathway in lactotrophs and the subsequent activation of inward rectifier K+ channels provide an effective and adenylyl cyclase-independent mechanism for a prolonged uncoupling of Ca2+ mobilization and influx pathways. C1 NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Stojilkovic, SS (reprint author), NICHHD, Sect Cellular Signaling, ERRB, Bldg 49,Room 6A-36,49 Convent Dr, Bethesda, MD 20892 USA. RI Tomic, Melanija/C-3371-2016 NR 44 TC 15 Z9 15 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD JUN PY 2002 VL 61 IS 6 BP 1329 EP 1339 DI 10.1124/mol.61.6.1329 PG 11 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LH UT WOS:000175736600009 PM 12021394 ER PT J AU Hoe, KL Saavedra, JM AF Hoe, KL Saavedra, JM TI Site-directed mutagenesis of the gerbil and human angiotensin II AT(1) receptors identifies amino acid residues attributable to the binding affinity for the nonpeptidic antagonist losartan SO MOLECULAR PHARMACOLOGY LA English DT Article ID 3RD TRANSMEMBRANE DOMAIN; MOLECULAR-CLONING; TYPE-1 RECEPTOR; EXPRESSION; RAT; PEPTIDE; BRAIN; ACTIVATION; MUTATION; SUBTYPE AB Gerbil angiotensin II AT(1) receptors have more than 90% amino acid sequence homology with human AT(1) receptors and similar affinity for the natural peptide agonist angiotensin II. However, their binding affinity for the biphenylimidazole AT(1) receptor antagonist losartan is greatly reduced compared with the hAT(1) receptor (400 times lower for the gAT(1A) receptor and 40 times lower for the gAT(1B) receptor cloned here). Gain- and loss-of-function site-directed mutagenesis revealed that in gerbil and human AT(1) receptors, the amino acid most important for losartan binding is located in position 108, followed by 107, both in transmembrane (TM) III. In both gerbil and human AT(1) receptors, the effect of G107S and I108V mutants is cumulative. Mutation L195M in TM V is very important, when combined with mutations G107S and I108V, for both gerbil and human AT1 receptors. In the gerbil, less important amino acids are located in positions 150/151 (TM IV) and 177 in the extracellular loop 2. The study of gerbil natural mutants allowed us to advance our understanding of amino acids selectively involved in the determination of antagonist affinity for gerbil and, most importantly, for human angiotensin II AT(1) receptors. C1 NIMH, Pharmacol Sect, Intramural Res Program, DIRP, Bethesda, MD 20892 USA. RP Saavedra, JM (reprint author), NIMH, Pharmacol Sect, Intramural Res Program, DIRP, 10 Ctr Dr,MSC 1514,Bldg 10,Room 2D-57, Bethesda, MD 20892 USA. NR 36 TC 10 Z9 10 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD JUN PY 2002 VL 61 IS 6 BP 1404 EP 1415 AR UNSP 1533/987785 DI 10.1124/mol.61.6.1404 PG 12 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LH UT WOS:000175736600017 PM 12021402 ER PT J AU Tokita, K Hocart, SJ Coy, DH Jensen, RT AF Tokita, K Hocart, SJ Coy, DH Jensen, RT TI Molecular basis of the selectivity of gastrin-releasing peptide receptor for gastrin-releasing peptide SO MOLECULAR PHARMACOLOGY LA English DT Article ID PROTEIN-COUPLED RECEPTOR; CATION-PI INTERACTIONS; MAMMALIAN BOMBESIN RECEPTORS; AFFINITY AGONIST BINDING; CENTRAL-NERVOUS-SYSTEM; NEUROMEDIN-B RECEPTOR; NEUROKININ-1 RECEPTOR; EXTRACELLULAR DOMAINS; TRANSMEMBRANE DOMAINS; AMINO-ACIDS AB The mammalian bombesin peptides [gastrin-releasing peptide (GRP) and neuromedin B (NMB)] are important in numerous biological and pathological processes. These effects are mediated by the heptahelical GRP receptor (GRPR) and NMB receptor (NMBR). GRP has high affinity for GRPR and lower affinity for NMBR. Almost nothing is known about the molecular basis for the selectivity of GRP. To address this question, we first studied four loss-of-affinity GRPR chimeric receptors formed by exchanging the four extracellular (EC) domains of GRPR with the corresponding NMBR EC domains. Receptors were transiently expressed, and affinities were determined by binding studies. Only substitution of the third EC domain (EC3) of GRPR markedly decreased GRP affinity. In the reverse study using gain-of-affinity NMBR chimeras, only replacement of EC3 of NMBR markedly increased GRP affinity. Replacing each of the 20 comparable EC3 amino acids that differed in the NMBR in GRPR showed that two separate NMBR substitutions in the GRPR, Ile for Phe(185) or Ile for Ala(198), markedly decreased GRP affinity. Additional point mutants demonstrated that an amino acid with an aromatic ring in position 185 of GRPR and the size of the backbone substitution in position 198 of GRPR were important for GRP selectivity. These results demonstrate that selectivity of GRP for GRPR over NMBR is primarily determined by two amino acid differences in the EC3 domains of the receptor. Our results suggest that an interaction between the aromatic ring of Phe(185) of the GRPR with GRP is the most important for GRP selectivity. C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. Tulane Univ, Hlth Sci Ctr, Dept Med, Peptide Res Labs, New Orleans, LA 70118 USA. RP Jensen, RT (reprint author), NIDDK, NIH, DDB, Bldg 10,Rm 9C-103,10 Ctr Dr,MSC 1804, Bethesda, MD 20892 USA. NR 43 TC 26 Z9 28 U1 0 U2 1 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD JUN PY 2002 VL 61 IS 6 BP 1435 EP 1443 AR UNSP 1472/985354 DI 10.1124/mol.61.6.1435 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 554LH UT WOS:000175736600020 PM 12021405 ER PT J AU Yordanov, AT Lodder, AL Woller, EK Cloninger, MJ Patronas, N Milenic, D Brechbiel, MW AF Yordanov, AT Lodder, AL Woller, EK Cloninger, MJ Patronas, N Milenic, D Brechbiel, MW TI Novel iodinated dendritic nanoparticles for computed tomography (CT) imaging SO NANO LETTERS LA English DT Article ID CONTRAST AGENTS; STARBURST(TM) DENDRIMERS; POLYETHYLENE-GLYCOL; CHEMISTRY; DELIVERY AB The design, synthesis, and characterization of novel water-soluble iodinated dendritic nanoparticles, G-4-(DMAA-IPA)(37), is reported. They consist of a generation 4 Starburst PAMAM dendrimer core and 3-N-[(N',N'-dimethylaminoacetyl)amino]-alpha-ethyl-2,4,6-triiodobenzenepropanoic acid (DMAA-IPA) molecules covalently attached to the surface. These nanoparticles have an average molecular weight of 37,000 g/mol, a hydrodynamic radius of 2.4 nm and high iodine content (33.06%), while retaining their overall charge and water solubility. Thus, G-4-(DMAA-IPA)(37) is the first representative of a novel class of dendritic nanoparticles for computed tomography (CT) imaging. C1 NIH, Radioimmune & Inorgan Chem Sect, Radiat Oncol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NIH, Dept Diagnost Radiol, Ctr Clin, Bethesda, MD 20892 USA. Wyatt Technol Corp, Santa Barbara, CA 93117 USA. Montana State Univ, Dept Chem & Biochem, Bozeman, MT 59717 USA. RP Yordanov, AT (reprint author), NIH, Radioimmune & Inorgan Chem Sect, Radiat Oncol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NR 30 TC 52 Z9 57 U1 1 U2 16 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 1530-6984 J9 NANO LETT JI Nano Lett. PD JUN PY 2002 VL 2 IS 6 BP 595 EP 599 DI 10.1021/nl020246x PG 5 WC Chemistry, Multidisciplinary; Chemistry, Physical; Nanoscience & Nanotechnology; Materials Science, Multidisciplinary; Physics, Applied; Physics, Condensed Matter SC Chemistry; Science & Technology - Other Topics; Materials Science; Physics GA 562KC UT WOS:000176195100008 ER PT J AU Tuan, R AF Tuan, R TI Boning up on telomerase SO NATURE BIOTECHNOLOGY LA English DT Editorial Material ID MESENCHYMAL STEM-CELLS C1 NIAMSD, Cartilage Biol & Orthopaed Branch, NIH, Bethesda, MD 20892 USA. RP Tuan, R (reprint author), NIAMSD, Cartilage Biol & Orthopaed Branch, NIH, Bethesda, MD 20892 USA. NR 10 TC 4 Z9 6 U1 0 U2 0 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1087-0156 J9 NAT BIOTECHNOL JI Nat. Biotechnol. PD JUN PY 2002 VL 20 IS 6 BP 560 EP 561 DI 10.1038/nbt0602-560 PG 2 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA 558NM UT WOS:000175973500019 PM 12042856 ER PT J AU Shi, ST Gronthos, S Chen, SQ Reddi, A Counter, CM Robey, PG Wang, CY AF Shi, ST Gronthos, S Chen, SQ Reddi, A Counter, CM Robey, PG Wang, CY TI Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression SO NATURE BIOTECHNOLOGY LA English DT Article ID IN-VIVO; CATALYTIC SUBUNIT; HUMAN FIBROBLASTS; POROUS HYDROXYAPATITE; REVERSE-TRANSCRIPTASE; IMMORTAL CELLS; LIFE-SPAN; CANCER; TISSUE; MICE AB Human postnatal bone marrow stromal stem cells (BMSSCs) have a limited life-span and progressively lose their stem cell properties during ex vivo expansion. Here we report that ectopic expression of human telomerase reverse transcriptase (hTERT) in BMSSCs extended their life-span and maintained their osteogenic potential. In xenogenic transplants, hTERT-expressing BMSSCs (BMSSC-Ts) generated more bone tissue, with a mineralized lamellar bone structure and associated marrow, than did control BMSSCs. The enhanced bone-forming ability of BMSSC-Ts was correlated with a higher and sustained expression of the early preosteogenic stem cell marker STRO-1, indicating that telomerase expression helped to maintain the osteogenic stem cell pool during ex vivo expansion. These results show that telomerase expression can overcome critical technical barriers to the ex vivo expansion of BMSSCs, and suggest that telomerase therapy may be a useful strategy for bone regeneration and repair. C1 Univ Michigan, Sch Dent, Dept Biol & Mat Sci, Lab Mol Signaling & Apoptosis, Ann Arbor, MI 48109 USA. Natl Inst Dent & Craniofacial Res, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. Duke Univ, Med Ctr, Dept Med, Durham, NC 27710 USA. RP Wang, CY (reprint author), Univ Michigan, Sch Dent, Dept Biol & Mat Sci, Lab Mol Signaling & Apoptosis, Ann Arbor, MI 48109 USA. RI Robey, Pamela/H-1429-2011; OI Robey, Pamela/0000-0002-5316-5576; Reddi, Anand/0000-0002-8360-8737 NR 47 TC 250 Z9 266 U1 0 U2 9 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1087-0156 J9 NAT BIOTECHNOL JI Nat. Biotechnol. PD JUN PY 2002 VL 20 IS 6 BP 587 EP 591 DI 10.1038/nbt0602-587 PG 5 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA 558NM UT WOS:000175973500024 PM 12042862 ER PT J AU Maynard, JA Maassen, CBM Leppla, SH Brasky, K Patterson, JL Iverson, BL Georgiou, G AF Maynard, JA Maassen, CBM Leppla, SH Brasky, K Patterson, JL Iverson, BL Georgiou, G TI Protection against anthrax toxin by recombinant antibody fragments correlates with antigen affinity SO NATURE BIOTECHNOLOGY LA English DT Article ID SINGLE-CHAIN FV; HUMANIZED MONOCLONAL-ANTIBODY; BACILLUS-ANTHRACIS; IN-VITRO; INHALATIONAL ANTHRAX; PHAGE DISPLAY; RECEPTOR; IMMUNITY; BINDING; VIVO AB The tripartite toxin produced by Bacillus anthracis is the key determinant in the etiology of anthrax. We have engineered a panel of toxin-neutralizing antibodies, including single-chain variable fragments (scFvs) and scFvs fused to a human constant domain (scAbs), that bind to the protective antigen subunit of the toxin with equilibrium dissociation constants (K-d) between 63 nM and 0.25 nM. The entire antibody panel showed high serum, thermal, and denaturant stability. In vitro, post-challenge protection of macrophages from the action of the holotoxin correlated with the K-d of the scFv variants. Strong correlations among antibody construct affinity, serum half-life, and protection were also observed in a rat model of toxin challenge. High-affinity toxin-neutralizing antibodies may be of therapeutic value for alleviating the symptoms of anthrax toxin in infected individuals and for medium-term prophylaxis to infection. C1 Univ Texas, Dept Chem Engn, Austin, TX 78712 USA. Univ Texas, Inst Cell & Mol Biol, Austin, TX 78712 USA. Univ Texas, Dept Chem & Biochem, Austin, TX 78712 USA. Univ Texas, Dept Biomed Engn, Austin, TX 78712 USA. Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD 20892 USA. SW Fdn Biol Res, San Antonio, TX 78227 USA. RP Georgiou, G (reprint author), Univ Texas, Dept Chem Engn, Austin, TX 78712 USA. NR 43 TC 196 Z9 202 U1 2 U2 18 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1087-0156 J9 NAT BIOTECHNOL JI Nat. Biotechnol. PD JUN PY 2002 VL 20 IS 6 BP 597 EP 601 DI 10.1038/nbt0602-597 PG 5 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA 558NM UT WOS:000175973500026 PM 12042864 ER PT J AU Bulavin, DV Demidov, ON Saito, S Kauraniemi, P Phillips, C Amundson, SA Ambrosino, C Sauter, G Nebreda, AR Anderson, CW Kallioniemi, A Fornace, AJ Appella, E AF Bulavin, DV Demidov, ON Saito, S Kauraniemi, P Phillips, C Amundson, SA Ambrosino, C Sauter, G Nebreda, AR Anderson, CW Kallioniemi, A Fornace, AJ Appella, E TI Amplification of PPM1D in human tumors abrogates p53 tumor-suppressor activity SO NATURE GENETICS LA English DT Article ID POSTTRANSLATIONAL MODIFICATIONS; PREMATURE SENESCENCE; GENETIC ALTERATIONS; HUMAN FIBROBLASTS; BREAST-CANCER; UV-RADIATION; P38 KINASE; PATHWAY; PROTEIN; PHOSPHATASE AB Expression of oncogenic Ras in primary human cells activates p53, thereby protecting cells from transformation. We show that in Ras-expressing IMR-90 cells, p53 is phosphorylated at Ser33 and Ser46 by the p38 mitogen-activated protein kinase (MAPK). Activity of p38 MAPK is regulated by the p53-inducible phosphatase PPM1D, creating a potential feedback loop. Expression of oncogenic Ras suppresses PPM1D mRNA induction, leaving p53 phosphorylated at Ser33 and Ser46 and in an active state. Retrovirus-mediated overexpression of PPM1D reduced p53 phosphorylation at these sites, abrogated Ras-induced apoptosis and partially rescued cells from cell-cycle arrest. Inactivation of p38 MAPK (the product of Mapk14) in vivo by gene targeting or by PPM1D overexpression expedited tumor formation after injection of mouse embryo fibroblasts (MEFs) expressing E1A+Ras into nude mice. The gene encoding PPM1D (PPM1D, at 17q22/q23) is amplified in human breast-tumor cell lines and in approximately 11% of primary breast tumors, most of which harbor wildtype p53. These findings suggest that inactivation of the p38 MAPK through PPM1D overexpression resulting from PPM1D amplification contributes to the development of human cancers by suppressing p53 activation. C1 NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. NIH, Gene Response Sect, Bethesda, MD 20892 USA. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. Tampere Univ, Inst Med Technol, Canc Genet Lab, Tampere, Finland. Tampere Univ Hosp, Tampere, Finland. European Mol Biol Lab, Heidelberg, Germany. Univ Basel, Inst Pathol, Basel, Switzerland. Brookhaven Natl Lab, Dept Biol, Upton, NY 11973 USA. RP Appella, E (reprint author), NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RI Fornace, Albert/A-7407-2008; OI Fornace, Albert/0000-0001-9695-085X; Demidov, Oleg/0000-0003-4323-7174; Kallioniemi, Anne/0000-0003-3552-8158 NR 31 TC 272 Z9 290 U1 2 U2 17 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD JUN PY 2002 VL 31 IS 2 BP 210 EP 215 DI 10.1038/ng894 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 557HR UT WOS:000175903500021 PM 12021785 ER PT J AU O'Shea, JJ Paul, WE AF O'Shea, JJ Paul, WE TI Regulation of THI differentiation - controlling the controllers SO NATURE IMMUNOLOGY LA English DT Editorial Material ID T-BET; COMMITMENT; STAT6 AB Although cytokines and STATs are accepted as being integral to T(H)1 differentiation, T-bet had emerged as a possible master control switch. Fresh data forces a reexamination of the factors that control T-H cell development. C1 NIAMSD, Mol Immunol & Inflammat Branch, NIH, Bethesda, MD 20892 USA. NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP O'Shea, JJ (reprint author), NIAMSD, Mol Immunol & Inflammat Branch, NIH, Bethesda, MD 20892 USA. NR 12 TC 58 Z9 59 U1 1 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 JUN PY 2002 VL 3 IS 6 BP 506 EP 508 DI 10.1038/ni0602-506 PG 4 WC Immunology SC Immunology GA 556XR UT WOS:000175874700006 PM 12032561 ER PT J AU Erman, B Feigenbaum, L Coligan, JE Singer, A AF Erman, B Feigenbaum, L Coligan, JE Singer, A TI Early TCR alpha expression generates TCR complexes that signal the DN-to-DP transition and impair development SO NATURE IMMUNOLOGY LA English DT Article ID DELTA-GENE REARRANGEMENT; CELL RECEPTOR-ALPHA; GAMMA-DELTA; TRANSGENIC MICE; THYMOCYTE DEVELOPMENT; MONOCLONAL-ANTIBODY; BETA EXPRESSION; CHAIN; SELECTION; DIFFERENTIATION AB Clonotypic T cell receptor (TCR) genes undergo ordered rearrangement and expression in the thymus with the result that TCRalpha and TCRgamma proteins are not expressed in the same cell at the same time. Such "TCRalpha/gamma exclusion" is a feature of normal thymocyte differentiation, but it is abrogated in TCR-transgenic mice, which prematurely express transgenic TCRalpha proteins in early double-negative (DN) thymocytes. We report here that early expression of TCRalpha proteins results in the formation of TCRalpha/gamma complexes that efficiently signal the differentiation of DN into double-positive thymocytes independently of pre-TCR and TCRbeta expression. Thus, abrogation of TCRalpha/gamma exclusion by early TCRalpha expression results in the formation of isotypically mixed TCRalphagamma complexes whose in vivo signals circumvent TCRbeta selection and redirect thymocyte development along an aberrant developmental pathway. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NCI, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NIAID, Lab Allerg Dis, Bethesda, MD 20892 USA. RP Singer, A (reprint author), NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NR 42 TC 24 Z9 24 U1 1 U2 1 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 JUN PY 2002 VL 3 IS 6 BP 564 EP 569 DI 10.1038/ni800 PG 6 WC Immunology SC Immunology GA 556XR UT WOS:000175874700016 PM 12021779 ER PT J AU Shih, TAY Meffre, E Roederer, M Nussenzweig, MC AF Shih, TAY Meffre, E Roederer, M Nussenzweig, MC TI Role of BCR affinity in T cell-dependent antibody responses in vivo SO NATURE IMMUNOLOGY LA English DT Article ID PRIMARY IMMUNE-RESPONSE; INFLUENZA-VIRUS HEMAGGLUTININ; CENTER B-CELLS; GERMINAL-CENTERS; SOMATIC HYPERMUTATION; PRIMARY IMMUNIZATION; SECONDARY RESPONSES; CLONAL SELECTION; MATURATION; MEMORY AB Antibody affinity for antigen is believed to govern B lymphocyte selection during T-dependent immune responses. To examine antibody affinity in T cell-dependent immune responses, we compared mice that carry targeted V(H)B1-8 antibody genes with high or low antigen-binding affinity. We found that high- and low-affinity B cells had the same intrinsic capacity to respond to antigen, but in experiments where limiting numbers of high- and low-affinity B cells were mixed in wild-type recipient mice, only the high- affinity B cells accumulated in germinal centers (GCs). In GCs, high-affinity B cells accumulated fewer V-H somatic mutations than low affinity B cells. This effect was due to selections as the frequency of mutation in noncoding immunoglobulin gene DNA is the same in high- and low-affinity B cells. Thus, B cells recruited to the GC appeared to undergo a fixed mutation program, regardless of initial B cell receptor affinity. We conclude that in addition to the selection that occurs in GCs, stringent selection for high- affinity clones is also imposed in the early stages of the T cell-dependent immune response in vivo. C1 Rockefeller Univ, Lab Mol Immunol, New York, NY 10021 USA. Rockefeller Univ, Howard Hughes Med Inst, New York, NY 10021 USA. NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. RP Nussenzweig, MC (reprint author), Rockefeller Univ, Lab Mol Immunol, 1230 York Ave, New York, NY 10021 USA. RI Roederer, Mario/G-1887-2011 FU NIGMS NIH HHS [GM07739] NR 50 TC 147 Z9 151 U1 0 U2 7 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 JUN PY 2002 VL 3 IS 6 BP 570 EP 575 DI 10.1038/ni803 PG 6 WC Immunology SC Immunology GA 556XR UT WOS:000175874700017 PM 12021782 ER PT J AU Kirk, AD AF Kirk, AD TI Location, location, location: Regional immune mechanisms critically influence rejection SO NATURE MEDICINE LA English DT Editorial Material C1 NIDDKD, Transplantat Sect, Transplantat & Autoimmun Branch, NIH, Bethesda, MD 20892 USA. RP Kirk, AD (reprint author), NIDDKD, Transplantat Sect, Transplantat & Autoimmun Branch, NIH, Bethesda, MD 20892 USA. RI Kirk, Allan/B-6905-2012 NR 10 TC 13 Z9 14 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 JUN PY 2002 VL 8 IS 6 BP 553 EP 555 DI 10.1038/nm0602-553 PG 3 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 557KG UT WOS:000175907300018 PM 12042797 ER PT J AU Coyle, JT Manji, HK AF Coyle, JT Manji, HK TI Getting balance: Drugs for bipolar disorder share target SO NATURE MEDICINE LA English DT Editorial Material ID LITHIUM; DEPRESSION; MECHANISM C1 Harvard Univ, Sch Med, McLean Dept Psychiat, Belmont, MA 02178 USA. NIMH, Bethesda, MD 20892 USA. RP Coyle, JT (reprint author), Harvard Univ, Sch Med, McLean Dept Psychiat, Belmont, MA 02178 USA. NR 11 TC 23 Z9 23 U1 0 U2 1 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 JUN PY 2002 VL 8 IS 6 BP 557 EP 558 DI 10.1038/nm0602-557 PG 2 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 557KG UT WOS:000175907300020 PM 12042799 ER PT J AU Landis, SC AF Landis, SC TI Quick-change artist: from excitatory to inhibitory synapse in minutes SO NATURE NEUROSCIENCE LA English DT Editorial Material ID RAT SYMPATHETIC NEURONS; CARDIAC MYOCYTES; NERVOUS-SYSTEM; DIFFERENTIATION; MICROCULTURES; ACETYLCHOLINE; TRANSMISSION; RELEASE AB Cultured sympathetic neurons contain an excitatory transmitter, norepinephrine, and one that is inhibitory, acetylcholine. A new paper shows that BDNF increases the ratio of acetylcholine to norepinephrine release, reversing the effect of neural stimulation from excitation to inhibition. C1 NINCDS, NIH, Bethesda, MD 20892 USA. RP Landis, SC (reprint author), NINCDS, NIH, Bldg 36,Rm 5A05,36 Convent Dr, Bethesda, MD 20892 USA. NR 15 TC 4 Z9 4 U1 0 U2 1 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 JUN PY 2002 VL 5 IS 6 BP 503 EP 504 DI 10.1038/nn0602-503 PG 2 WC Neurosciences SC Neurosciences & Neurology GA 556XQ UT WOS:000175874600002 PM 12037514 ER PT J AU Shevach, EM AF Shevach, EM TI CD4(+)CD25(+) suppressor T cells: More questions than answers SO NATURE REVIEWS IMMUNOLOGY LA English DT Review ID IMMUNOLOGICAL SELF-TOLERANCE; GROWTH-FACTOR-BETA; SPONTANEOUS AUTOIMMUNE ENCEPHALOMYELITIS; LYMPHOCYTE-ASSOCIATED ANTIGEN-4; ORGAN-SPECIFIC AUTOIMMUNITY; HUMAN PERIPHERAL-BLOOD; REGULATORY CELLS; DENDRITIC CELLS; IN-VITRO; TGF-BETA AB Several mechanisms control discrimination between self and non-self, including the thymic deletion of autoreactive T cells and the induction of anergy in the periphery. In addition to these passive mechanisms, evidence has accumulated for the active suppression of autoreactivity by a population of regulatory or suppressor T cells that co-express CD4 and CD25 (the interleukin-2 receptor alpha-chain). CD4(+)CD25(+) T cells are powerful inhibitors of T-cell activation both in vivo and in vitro. The enhancement of suppressor-cell function might prove useful for the treatment of immune-mediated diseases, whereas the downregulation of these cells might be beneficial for the enhancement of the immunogenicity of vaccines that are specific for tumour antigens. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Shevach, EM (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Room 11N315, Bethesda, MD 20892 USA. RI Yang, Chen/G-1379-2010; Liu, Xia/L-9425-2013 NR 85 TC 1606 Z9 1709 U1 1 U2 38 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1474-1733 J9 NAT REV IMMUNOL JI Nat. Rev. Immunol. PD JUN PY 2002 VL 2 IS 6 BP 389 EP 400 DI 10.1038/nri821 PG 12 WC Immunology SC Immunology GA 636DE UT WOS:000180438600018 PM 12093005 ER PT J AU Maurizi, MR AF Maurizi, MR TI Love it or cleave it: Tough choices in protein quality control SO NATURE STRUCTURAL BIOLOGY LA English DT Editorial Material ID SERINE-PROTEASE; MITOCHONDRIA; HTRA2 AB Housekeeping is not a humdrum process when it comes to protein quality control. Inside chaperone-protease machines, proteins are subject to dramatic life or death trials and must shape up rapidly to survive. Accumulation of aberrantly folded proteins is the basis of many diseases, so the outcome is vitally important to the cell. C1 NCI, Cell Biol Lab, Bethesda, MD 20892 USA. RP Maurizi, MR (reprint author), NCI, Cell Biol Lab, Bldg 37, Bethesda, MD 20892 USA. NR 12 TC 12 Z9 13 U1 0 U2 0 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1072-8368 J9 NAT STRUCT BIOL JI Nat. Struct. Biol. PD JUN PY 2002 VL 9 IS 6 BP 410 EP 412 DI 10.1038/nsb0602-410 PG 4 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 556XP UT WOS:000175874500004 PM 12032552 ER PT J AU Abbott, KC Agodoa, LY AF Abbott, KC Agodoa, LY TI Hospitalizations for bacterial endocarditis after initiation of chronic dialysis in the United States SO NEPHRON LA English DT Article DE bacterial endocarditis; hospitalization; complications; dialysis; United States Renal Data System; age; creatinine; African-American; diabetes; nonvalvular heart disease ID STAGE RENAL-DISEASE; INFECTIVE ENDOCARDITIS; RISK-FACTORS; STAPHYLOCOCCUS-AUREUS; CHRONIC-HEMODIALYSIS; BACTEREMIA; TRANSPLANT; PROGNOSIS; MORTALITY; EPISODES AB Aims: Bacterial endocarditis is a significant cause of morbidity and mortality but has not been studied in a national population of end-stage renal disease patients. Methods: 327,993 dialysis patients in the United States Renal Data System initiated from 1 January 1992 to 30 June 1997 were analyzed in a historical cohort study of hospitalized bacterial endocarditis (ENDO, ICD9 Code 421.x). Renal transplant recipients were excluded. Results: Hemodialysis patients had an age-adjusted incidence ratio for ENDO of 17.86 (95% confidence interval, 6.62-48.90) and peritoneal dialysis patients 10.54 (95% Cl, 0.71- 158.13, not statistically significant) compared to the general population in 1996 (the National Hospital Discharge Survey). 6.1% of patients with ENDO underwent valve replacement surgery. In multivariate analysis, hemodialysis (vs. peritoneal dialysis), earlier year of dialysis, cardiac disease, and lower serum creatinine and albumin were associated with increased risk of ENDO. In Cox regression analysis, patients with ENDO had increased mortality, relative risk 1.48 (95% Cl 1.45-1.73). Conclusions: Patients on chronic dialysis were at increased risk for ENDO compared to the general population. The risk for peritoneal dialysis patients was not statistically significant, possibly due to the smaller numbers of patients on this modality. Hemodialysis (vs. peritoneal dialysis) and comorbidities were the strongest risk factors for ENDO identified. Copyright (C) 2002 S. Karger AG, Basel. C1 Walter Reed Army Med Ctr, Serv Nephrol, Washington, DC 20307 USA. Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA. NIDDK, NIH, Bethesda, MD USA. RP Abbott, KC (reprint author), Walter Reed Army Med Ctr, Serv Nephrol, Washington, DC 20307 USA. OI Abbott, Kevin/0000-0003-2111-7112 NR 30 TC 49 Z9 50 U1 0 U2 2 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 0028-2766 J9 NEPHRON JI Nephron PD JUN PY 2002 VL 91 IS 2 BP 203 EP 209 DI 10.1159/000058393 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA 566EV UT WOS:000176414800005 PM 12053054 ER PT J AU Dreher, JC Burnod, Y AF Dreher, JC Burnod, Y TI An integrative theory of the phasic and tonic modes of dopamine modulation in the prefrontal cortex SO NEURAL NETWORKS LA English DT Review DE dopamine; inverted U-curve; D1 receptors; working memory; prefrontal cortex ID VENTRAL TEGMENTAL AREA; VISUOSPATIAL WORKING-MEMORY; DELAYED-RESPONSE TASK; NUCLEUS-ACCUMBENS; PYRAMIDAL NEURONS; EXTRACELLULAR DOPAMINE; COGNITIVE PERFORMANCE; INHIBITORY INFLUENCE; PRELIMBIC CORTEX; CORTICAL-NEURONS AB This paper presents a model of both tonic and phasic dopamine (DA) effects on maintenance of working memory representations in the prefrontal cortex (PFC). The central hypothesis is that DA modulates the efficacy of inputs to prefrontal pyramidal neurons to prevent interferences for active maintenance. Phasic DA release, due to DA neurons discharges, acts at a short time-scale (a few seconds), while the tonic mode of DA release, independent of DA neurons firing, acts at a long time-scale (a few minutes). The overall effect of DA modulation is modeled as a threshold restricting incoming inputs arriving on PFC neurons. Phasic DA release temporary increases this threshold while tonic DA release progressively increases the basal level of this threshold. Thus, unlike the previous gating theory of phasic DA release, proposing that it facilitates incoming inputs at the time of their arrival, the effect of phasic DA release is supposed to restrict incoming inputs during a period of time after DA neuron discharges. The model links the cellular and behavioral levels during performance of a working memory task. It allows us to understand why a critical range of DA D1 receptors stimulation is required for optimal working memory performance and how D1 receptor agonists (respectively antagonists) increase perseverations (respectively distractability). Finally, the model leads to several testable predictions, including that the PFC regulates DA neurons firing rate to adapt to the delay of the task and that increase in tonic DA release may either improve or decrease performance, depending on the level of DA receptors stimulation at the beginning of the task. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Univ Paris 06, INSERM, U 483, F-75005 Paris, France. RP Dreher, JC (reprint author), NIMH, Clin Brain Disorder Branch, Room 4C-108,MSC 1440, Bethesda, MD 20892 USA. NR 100 TC 39 Z9 39 U1 0 U2 5 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0893-6080 J9 NEURAL NETWORKS JI Neural Netw. PD JUN-JUL PY 2002 VL 15 IS 4-6 BP 583 EP 602 AR PII S0893-6080(02)00051-5 DI 10.1016/S0893-6080(02)00051-5 PG 20 WC Computer Science, Artificial Intelligence SC Computer Science GA 594ZZ UT WOS:000178083500010 PM 12371514 ER PT J AU Mead, AN Katz, JL Rocha, BA AF Mead, AN Katz, JL Rocha, BA TI Intravenous cocaine-induced activity in A/J and C57BL/6J mice: behavioral sensitization and conditioned activity SO NEUROPHARMACOLOGY LA English DT Article DE habituation hypothesis; conditioning; mouse; locomotor activity ID LOCOMOTOR-ACTIVITY; INCENTIVE-SENSITIZATION; PLACE PREFERENCE; AMPHETAMINE; RATS; DOPAMINE; RECEPTORS; ACTIVATION; ADDICTION; INTRAPERITONEAL AB The stimulant properties of cocaine have been extensively investigated in the mouse,C using either intraperitoneal (i.p.) or subcutaneous (s.c.) administration of drug. However. cocaine use in humans often invokes intravenous,11, (i.v.) administration of drug. The purpose of this study was to develop a methodology for studying cocaine-induced activity in the mouse. which allows within session determination of the dose-response function. and assessment of the development of behavioral sensitization and conditioned activity. The stimulant effects of i.v. cocaine (3-25 mg/kg) were investigated in C57BL/6J and A/J mice both acutely and following repeated treatments (four treatments at 48 hour intervals). in addition to the conditioned activating properties of I the cocaine-paired context. Cocaine produced a dose-dependent increase in measures of motor activity in both strains of mice. Repeated cocaine treatments resulted in the development of behavioral sensitization to the stimulant properties of the drug at 111 doses tested, and exposure to the cocaine-paired context in the absence of drug revealed the development of conditioned activity, While both C57BL/6J and A/J strains displayed these phenomena, differences were observed between ambulation and total beam break,,, highlighting differences between multiple behavioral end-points. Both strains of mice displayed conditioned activity Of a higher magnitude than their response to novelty. in addition to a positive relationship between the number of drug-environment pairings and the magnitude of the conditioned response. In summary. these data extend to the in route of administration previous, observations cri cocaine-induced activity and conditioned activity. Published by Elsevier Science Ltd. C1 Natl Inst Drug Abuse, Intramural Res Program, Behav Neurosci Branch, Baltimore, MD 21222 USA. Natl Inst Drug Abuse, Intramural Res Program, Medicat Discovery Res Branch, Baltimore, MD 21222 USA. Univ Maryland, Maryland Psychiat Res Ctr, Catonsville, MD 21228 USA. RP Mead, AN (reprint author), Univ Sussex, Lab Expt Psychol, Sch Biol Sci, Brighton BN1 9QG, E Sussex, England. OI Katz, Jonathan/0000-0002-1068-1159 NR 38 TC 11 Z9 11 U1 0 U2 7 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0028-3908 J9 NEUROPHARMACOLOGY JI Neuropharmacology PD JUN PY 2002 VL 42 IS 7 BP 976 EP 986 AR PII S0028-3908(02)00048-5 DI 10.1016/S0028-3908(02)00048-5 PG 11 WC Neurosciences; Pharmacology & Pharmacy SC Neurosciences & Neurology; Pharmacology & Pharmacy GA 577WX UT WOS:000177086600011 PM 12069908 ER PT J AU Matsumoto, RR McCracken, KA Pouw, B Zhang, Y Bowen, WD AF Matsumoto, RR McCracken, KA Pouw, B Zhang, Y Bowen, WD TI Involvement of sigma receptors in the behavioral effects of cocaine: evidence from novel ligands and antisense oligodeoxynucleotides SO NEUROPHARMACOLOGY LA English DT Article DE antisense; cocaine; convulsions; lethality; locomotor; sigma receptor ID CONDITIONED PLACE PREFERENCE; TRANSPORTER KNOCKOUT MICE; DOPAMINE-TRANSPORTER; INDUCED CONVULSIONS; RAT-BRAIN; MOLECULAR-CLONING; BINDING; STIMULATION; ANTAGONISM; SEROTONIN AB Pharmacological and molecular biological tools were used to validate the involvement of sigma receptors in the actions of cocaine. Radioligand binding studies demonstrated significant levels of sigma receptors in the brain and heart, where cocaine interacts preferentially with the sigma(1) subtype. In behavioral pharmacological studies using mice, nine novel sigma receptor antagonists significantly attenuated cocaine-induced convulsions, while structural analogs with weak interactions with sigma receptors were ineffective. In contrast to the protection provided by the antagonists, a classical sigma receptor agonist exacerbated the convulsive effects of cocaine. The antagonists also attenuated cocaine-induced lethality, with the best compound protecting against death even when administered as a post-treatment. At doses where the antagonists had no effect on baseline locomotor activity, they significantly attenuated the locomotor stimulatory effects of cocaine, suggesting their ability to block the psychomotor as well as the toxic effects of cocaine. To further validate that the anti-cocaine effects were achieved by interfering with cocaine's access to sigma receptors, antisense oligodeoxynucleotides against sigma(1) receptors were shown to attenuate the convulsive and locomotor stimulatory effects of cocaine. Together, the studies support the involvement of sigma receptors, particularly the sigma(1), subtype, in the behavioral effects of cocaine. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Univ Oklahoma, Hlth Sci Ctr, Dept Pharmaceut Sci, Oklahoma City, OK 73190 USA. NIDDK, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Matsumoto, RR (reprint author), Univ Oklahoma, Hlth Sci Ctr, Dept Pharmaceut Sci, POB 26901, Oklahoma City, OK 73190 USA. FU NIDA NIH HHS [DA 11979]; NIMH NIH HHS [MH 50564] NR 48 TC 67 Z9 68 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0028-3908 J9 NEUROPHARMACOLOGY JI Neuropharmacology PD JUN PY 2002 VL 42 IS 8 BP 1043 EP 1055 AR PII S0028-3908(02)00056-4 DI 10.1016/S0028-3908(02)00056-4 PG 13 WC Neurosciences; Pharmacology & Pharmacy SC Neurosciences & Neurology; Pharmacology & Pharmacy GA 584TW UT WOS:000177485700007 PM 12128006 ER PT J AU Gimenez-Llort, L Wang, FH Ogren, SO Ferre, S AF Gimenez-Llort, L Wang, FH Ogren, SO Ferre, S TI Local dopaminergic modulation of the motor activity induced by N-methyl-D-aspartate receptor stimulation in the ventral hippocampus SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE glutamate; dopamine D1 receptor; dopamine D2 receptor; general motor activity; locomotor activity; rearing ID NUCLEUS-ACCUMBENS; LOCOMOTOR-ACTIVITY; TEGMENTAL AREA; BASAL GANGLIA; MOUSE-BRAIN; RAT; MICE; D-1; POTENTIATION; EXPRESSION AB Dopaminergic neurotransmission has been implicated in the motor activating effects induced by the local infusion of NMDA in the ventral hippocampus (VH). The nucleus accumbens and the ventral tegmental area (VTA) have been proposed to be the main loci where dopamine is acting as a positive modulator of the VH NMDA receptor-mediated motor activating effects. However, the existence of a relatively high dopamine innervation and dopamine receptor density in the VH suggests the possibility of local dopamine/NMDA receptor interactions. this hypothesis was tested by studying the effects of the bilateral local VH infusion of NMDA (0.05, 0.1, 0.5 and 1.0 mug/side), the dopamine D1/D5 receptor antagonist SCH 23390 (1 mug/side) and the dopamine D2 receptor antagonist raclopride (1 and 5 mug/side). Neither SCH 23390 nor raclopride induced any significant change in motor activity compared with the vehicle control group, but both compounds significantly antagonized the motor activation induced by NMDA. SCH 23390 (1 mug/side) was more potent that raclopride (minimal effective dose: 5 mug/side). These results demonstrate the existence of a strong tonic facilitating effect of dopamine, acting preferentially at dopamine D1/D5 receptors, on NMDA receptor-mediated effects in the VH. (C) 2002 American College of Neuropsychopharmacology. Published by Elsevier Science Inc. C1 NIDA, Behav Neurosci Branch, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Karolinska Inst, Dept Neurosci, S-17177 Stockholm, Sweden. Autonomous Univ Barcelona, Sch Med, Dept Psychiat & Forens Med, E-08193 Barcelona, Spain. RP Ferre, S (reprint author), NIDA, Behav Neurosci Branch, Intramural Res Program, NIH, 5500 Nathan Shock Dr,POB 5180, Baltimore, MD 21224 USA. RI Ferre, Sergi/K-6115-2014; OI Ferre, Sergi/0000-0002-1747-1779; Ogren, Sven Ove/0000-0003-2573-5276 NR 25 TC 16 Z9 16 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD JUN PY 2002 VL 26 IS 6 BP 737 EP 743 AR PII S0893-133X(01)00411-0 DI 10.1016/S0893-133X(01)00411-0 PG 7 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 552RD UT WOS:000175632600004 PM 12007744 ER PT J AU Broom, DC Jutkiewicz, EM Folk, JE Traynor, JR Rice, KC Woods, JH AF Broom, DC Jutkiewicz, EM Folk, JE Traynor, JR Rice, KC Woods, JH TI Nonpeptidic delta-opioid receptor agonists reduce immobility in the forced swim assay in rats SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE delta opioid receptor; depression; forced swim assay; SNC80; (+)BW373U86 ID ANTIDEPRESSANT ACTIVITY; ENDOGENOUS ENKEPHALINS; MICE; BW373U86; SNC-80; MODEL; IMMUNOREACTIVITY; STIMULATION; PLASMA AB The present study examined the effect of opioid receptor agonists in the rat forced swim assay. The delta-opioid receptor agonists SNC80 ((+)-4-[(aR)-alpha-((2S,5R)-4-Allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N- diethylbenzamide) and (+)BW373U86 ((+)-[1(S*),2alpha,5beta]-4-[[2,5-dimethyl-4-(2-propenyl)-1-piperazinyl] (3-hydroxyphenyl)methyl]-N,N-diethyl-benzamide dihydrochloride) produced a decrease in immobility indicating an antidepressant-like effect. At antinociceptive doses, neither the kappa-opioid selective agonist C1977 (5R-(5alpha,7alpha,8beta)-N-methyl-N-[7-(1-pyrrolidinyl-1- oxaspiro[4,5]dec-8-yl]-4-benzofuranacetamide) showed a change in immobility that was identifiable by dose, nor were changes in immobility seen with morphine. A delta-opioid mechanism of action in the forced swim assail was likely since naltrindole prevented the effects of both delta-agonists. When compared to desipramine and fluoxetine, SNC80 was more active with a single dose whereas both desipramine and fluoxetine produced greater effects with subchronic dosing (3 doses). All three compounds were active when administered before the initial swim exposure. SNC80 was, however, more effective following a single dose than by subchronic administration demonstrating both a fast onset of activity and potential tolerance, Thus, delta-agonists differ from typical antidepressants in the forced swim assay. (C) 2002 American College of Neuropsychopharmacology. Published by Elsevier Science Inc. C1 Univ Michigan, Sch Med, Dept Pharmacol, Ann Arbor, MI 48109 USA. Univ Michigan, Sch Med, Dept Psychol, Ann Arbor, MI 48109 USA. NIDDK, Bethesda, MD 20892 USA. RP Woods, JH (reprint author), Univ Michigan, Sch Med, Dept Pharmacol, 1301 MSRBIII, Ann Arbor, MI 48109 USA. FU NIDA NIH HHS [DA00254, DA07267]; NIGMS NIH HHS [GM07767] NR 27 TC 106 Z9 107 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD JUN PY 2002 VL 26 IS 6 BP 744 EP 755 AR PII S0893-133X(01)00413-4 DI 10.1016/S0893-133X(01)00413-4 PG 12 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 552RD UT WOS:000175632600005 PM 12007745 ER PT J AU Hallett, M AF Hallett, M TI Recent advances in stroke rehabilitation SO NEUROREHABILITATION AND NEURAL REPAIR LA English DT Article ID INDUCED MOVEMENT THERAPY; POSITRON-EMISSION-TOMOGRAPHY; PERIPHERAL-NERVE STIMULATION; HUMAN MOTOR CORTEX; PHYSICAL-THERAPY; DOUBLE-BLIND; NORADRENERGIC PHARMACOTHERAPY; ELECTRICAL-STIMULATION; MUSCLE RESPONSES; COMMON-DRUGS 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. EM hallettm@ninds.nih.gov NR 66 TC 18 Z9 20 U1 0 U2 1 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1545-9683 J9 NEUROREHAB NEURAL RE JI Neurorehabil. Neural Repair PD JUN PY 2002 VL 16 IS 2 BP 211 EP 217 DI 10.1177/0888439002016002004 PG 7 WC Clinical Neurology; Rehabilitation SC Neurosciences & Neurology; Rehabilitation GA 845IH UT WOS:000223235200004 PM 15151118 ER PT J AU Nikolskaya, AN Galperin, MY AF Nikolskaya, AN Galperin, MY TI A novel type of conserved DNA-binding domain in the transcriptional regulators of the AlgR/AgrA/LytR family SO NUCLEIC ACIDS RESEARCH LA English DT Article ID 2-COMPONENT SIGNAL-TRANSDUCTION; LACTOBACILLUS-PLANTARUM C-11; RESPONSE REGULATOR; STREPTOCOCCUS-PNEUMONIAE; MOLECULAR CHARACTERIZATION; CLOSTRIDIUM-PERFRINGENS; PSEUDOMONAS-AERUGINOSA; BACTERIOCIN PRODUCTION; STAPHYLOCOCCUS-AUREUS; ALGD PROMOTER AB Sequence analysis of bacterial genomes revealed a novel DNA-binding domain. This domain is found in several response regulators of the two-component signal transduction system, such as Pseudomonas aeruginosa AlgR, involved in the regulation of alginate biosynthesis and in the pathogenesis of cystic fibrosis; Clostridium perfringens VirR, a regulator of virulence factors, and in several regulators of bacteriocin biosynthesis, previously unified in the AgrA/ComE family. Most of the transcriptional regulators that contain this DNA-binding domain are involved in biosynthesis of extracellular polysaccharides, fimbriation, expression of exoproteins, including toxins, and quorum sensing. We refer to it as the LytTR ('litter') domain, after Bacillus subtilis LytT and Staphylococcus aureus LytR response regulators, involved in regulation of cell autolysis. In addition to response regulators, the LytTR domain is found in combination with MHYT, PAS and other sensor domains. C1 Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RP Galperin, MY (reprint author), Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RI Galperin, Michael/B-5859-2013 OI Galperin, Michael/0000-0002-2265-5572 NR 36 TC 102 Z9 110 U1 1 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 JUN 1 PY 2002 VL 30 IS 11 BP 2453 EP 2459 DI 10.1093/nar/30.11.2453 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 563LE UT WOS:000176256500022 PM 12034833 ER PT J AU Shevchenko, Y Bouffard, GG Butterfield, YSN Blakesley, RW Hartley, JL Young, AC Marra, MA Jones, SJM Touchman, JW Green, ED AF Shevchenko, Y Bouffard, GG Butterfield, YSN Blakesley, RW Hartley, JL Young, AC Marra, MA Jones, SJM Touchman, JW Green, ED TI Systematic sequencing of cDNA clones using the transposon Tn5 SO NUCLEIC ACIDS RESEARCH LA English DT Article ID IN-VITRO; HUMAN GENOME; DNA; STRATEGY; TAGS; COLLECTION; DROSOPHILA; CLONING; GENES; TOOL AB In parallel with the production of genomic sequence data, attention is being focused on the generation of comprehensive cDNA-sequence resources. Such efforts are increasingly emphasizing the production of high-accuracy sequence corresponding to the entire insert of cDNA clones, especially those presumed to reflect the full-length mRNA. The complete sequencing of cDNA clones on a large scale presents unique challenges because of the generally small, yet heterogeneous, sizes of the cloned inserts. We have developed a strategy for high-throughput sequencing of cDNA clones using the transposon Tn5. This approach has been tailored for implementation within an existing large-scale 'shotgun-style' sequencing program, although it could be readily adapted for use in virtually any sequencing environment. In addition, we have developed a modified version of our strategy that can be applied to cDNA clones with large cloning vectors, thereby overcoming a potential limitation of transposon-based approaches. Here we describe the details of our cDNA-sequencing pipeline, including a summary of the experience in sequencing more than 4200 cDNA clones to produce more than 8 million base pairs of high-accuracy cDNA sequence. These data provide both convincing evidence that the insertion of Tn5 into cDNA clones is sufficiently random for its effective use in large-scale cDNA sequencing as well as interesting insight about the sequence context preferred for insertion by Tn5. C1 NIH, Intramural Sequencing Ctr, Gaithersburg, MD 20877 USA. NHGRI, Genome Technol Branch, Bethesda, MD 20892 USA. British Columbia Canc Agcy, Genome Sci Ctr, Vancouver, BC V5Z 4E6, Canada. Invitrogen Corp, Rockville, MD 20850 USA. RP NIH, Intramural Sequencing Ctr, 50 South Dr,Bldg 50,Room 5523, Gaithersburg, MD 20877 USA. EM egreen@nhgri.nih.gov RI Tang, Macy/B-9798-2014; Jones, Steven/C-3621-2009; Marra, Marco/B-5987-2008 NR 34 TC 41 Z9 43 U1 1 U2 5 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 EI 1362-4962 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD JUN 1 PY 2002 VL 30 IS 11 BP 2469 EP 2477 DI 10.1093/nar/30.11.2469 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 563LE UT WOS:000176256500024 PM 12034835 ER PT J AU Rapic-Otrin, V McLenigan, MP Bisi, DC Gonzalez, M Levine, AS AF Rapic-Otrin, V McLenigan, MP Bisi, DC Gonzalez, M Levine, AS TI Sequential binding of UV DNA damage binding factor and degradation of the p48 subunit as early events after UV irradiation SO NUCLEIC ACIDS RESEARCH LA English DT Article ID NUCLEOTIDE EXCISION-REPAIR; PIGMENTOSUM GROUP-E; RNA-POLYMERASE-II; XERODERMA-PIGMENTOSUM; PROTEIN DDB; ULTRAVIOLET PHOTOPRODUCTS; HUMAN FIBROBLASTS; PYRIMIDINE DIMER; MAMMALIAN-CELLS; PRIMATE CELLS AB The UV-damaged DNA binding protein complex (UV-DDB) is implicated in global genomic nucleotide excision repair (NER) in mammalian cells. The complex consists of a heterodimer of p127 and p48. UV-DDB is defective in one complementation group (XP-E) of the heritable, skin cancer-prone disorder xeroderma pigmentosum. Upon UV irradiation of primate cells, UV-DDB associates tightly with chromatin, concomitant with the loss of extractable binding activity. We report here that an early event after UV, but not ionizing, radiation is the transient dose-dependent degradation of the small subunit, p48. Treatment of human cells with the proteasomal inhibitor NIP-L3VS blocks this UV-induced degradation of p48. In XP-E cell lines with impaired UV-DDB binding, p48 is resistant to degradation. UV-mediated degradation of p48 occurs independently of the expression of p53 and the cell's proficiency for NER, but recovery of p48 levels at later times (12 h and thereafter) is dependent upon the capacity of the cell to repair non-transcribed DNA. In addition, we find that the p127 subunit of UV-DDB binds in vivo to p300, a histone acetyltransferase. The data support a functional connection between UV-DDB binding activity, proteasomal degradation of p48 and chromatin remodeling during early steps of NER. C1 Univ Pittsburgh, Sch Med, Dept Mol Genet & Biochem, Pittsburgh, PA 15261 USA. NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bethesda, MD 20892 USA. Univ Colorado, Dept Biol, Denver, CO 80217 USA. RP Levine, AS (reprint author), Univ Pittsburgh, Sch Med, Dept Mol Genet & Biochem, E1240 BST, Pittsburgh, PA 15261 USA. NR 67 TC 116 Z9 119 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 JUN 1 PY 2002 VL 30 IS 11 BP 2588 EP 2598 DI 10.1093/nar/30.11.2588 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 563LE UT WOS:000176256500037 PM 12034848 ER PT J AU Jamieson, DJ Kaufman, SC Costello, C Hillis, SD Marchbanks, PA Peterson, HB Hughes, JM Xia, ZS Wilcox, LS Tylor, LR Trussell, J Courey, NG Darney, PD Friedrich, ER Hale, RW Nakayama, RT Hulka, JF Poindexter, AN Ryan, GM Thorpe, EM Stewart, GK Zacur, HA Blanco, L AF Jamieson, DJ Kaufman, SC Costello, C Hillis, SD Marchbanks, PA Peterson, HB Hughes, JM Xia, ZS Wilcox, LS Tylor, LR Trussell, J Courey, NG Darney, PD Friedrich, ER Hale, RW Nakayama, RT Hulka, JF Poindexter, AN Ryan, GM Thorpe, EM Stewart, GK Zacur, HA Blanco, L CA US Collaborative Review Sterilizat TI A comparison of women's regret after vasectomy versus tubal sterilization SO OBSTETRICS AND GYNECOLOGY LA English DT Article ID POSTSTERILIZATION REGRET; UNITED-STATES; MEN AB OBJECTIVE: To compare the 5-year cumulative probability of regret and risk factors for regret among women whose husbands underwent vasectomy with women after tubal sterilization. METHODS. A total of 525 women whose husbands underwent vasectomy were compared with 3672 women who underwent tubal sterilization in a prospective, multicenter, cohort study. RESULTS: The cumulative probability of a woman expressing regret within 5 years after her husband's vasectomy was 6.1% (95% confidence interval [CI] 3.6, 8.6), which was similar to the 5-year cumulative probability of regret among women after tubal sterilization (7.0%,95% CI 5.8, 8.1). Women who reported substantial conflict with their husbands before vasectomy were more than 25 times more likely to request that their husband have a reversal than women who did not report such conflict (rate ratio 25.3, 95% CI 2.9, 217.2). Similarly, women who reported substantial conflict with their husbands or partners before tubal sterilization were more then three times as likely to regret their decision and more than five times as likely to request a reversal than women who did not report such conflict (rate ratio 3.1, 95% CI 1.4, 7.0, and rate ratio 5.4, 95% CI 1.6, 17.6, respectively). CONCLUSION: Most women did not express regret after their husband's vasectomy and the probability of regret was similar to sterilized women. However, when there was substantial conflict between a woman and her husband before vasectomy or tubal sterilization, the probability of subsequent request for reversal was increased. (Obstet Gynecol 2002;99:1073-9. (C) 2002 by the American College of Obstetricians and Gynecologists). C1 CDCP, Div Reprod Hlth, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA 30333 USA. NICHHD, Contracept & Reprod Hlth Branch, Populat Res Ctr, NIH, Bethesda, MD 20892 USA. RP Jamieson, DJ (reprint author), CDCP, Div Reprod Hlth, Natl Ctr Chron Dis Prevent & Hlth Promot, Mailstop E-45,1600 Clifton Rd, Atlanta, GA 30333 USA. FU NICHD NIH HHS [3-Y02-HD41075-10] NR 10 TC 32 Z9 35 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD JUN PY 2002 VL 99 IS 6 BP 1073 EP 1079 AR PII S0029-7844(02)01981-6 DI 10.1016/S0029-7844(02)01981-6 PG 7 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 558QF UT WOS:000175977500022 PM 12052602 ER PT J AU Anderson, J Rappoport, CA Hanson, C Maupin, R Minkoff, H O'Sullivan, MJ Perryman, S Scott, G Spector, SA Tuomala, R Wade, N Whitley-Williams, P Wilfert, C Zorrilla, C McNamara, J Mofenson, L Watts, DH Fowler, MG Jamieson, DJ Barini-Garcia, M Hench, K Baylor, M Cargill, VA AF Anderson, J Rappoport, CA Hanson, C Maupin, R Minkoff, H O'Sullivan, MJ Perryman, S Scott, G Spector, SA Tuomala, R Wade, N Whitley-Williams, P Wilfert, C Zorrilla, C McNamara, J Mofenson, L Watts, DH Fowler, MG Jamieson, DJ Barini-Garcia, M Hench, K Baylor, M Cargill, VA CA Public Hlth Serv Task Force Perina TI Summary of the updated recommendations from the public health service task force to reduce perinatal human immunodeficiency virus-1 transmission in the United States SO OBSTETRICS AND GYNECOLOGY LA English DT Editorial Material ID INFECTED WOMEN; ZIDOVUDINE; TYPE-1; INFANTS; THERAPY; EXPOSURE; TOXICITY; DELIVERY; MODE AB Within the last decade, substantial advances have been made in the treatment of human immunodeficiency virus (HIV)-infected pregnant women and in the prevention of perinatal HIV-1 transmission, and recommendations for care continually change. Within this rapidly evolving field, the Public Health Service Task Force Perinatal HIV Guidelines Working Group, which is composed of pediatric and obstetric experts in HIV infection, community representatives, and federal agency representatives, currently meets by monthly conference calls to review new data related to prevention of mother-to-child HIV transmission and management of women with HIV infection. This group periodically issues updates to their guidelines, "Public Health Service Task Force Recommendations for Use of Antiretroviral Drugs in Pregnant HIV-1-Infected Women for Maternal Health and Interventions to Reduce Perinatal HIV-1 Transmission in the United States," which are available on the HIV/AIDS Treatment Information Service Web site (http://www.hivatis.org). (Obstet Gynecol 2002;99: 1117-26. (C) 2002 by the American College of Obstetricians and Gynecologists). C1 CDCP, Div HIV AIDS, Natl Ctr HIV STD & TB Prevent, Atlanta, GA 30333 USA. Johns Hopkins Univ, Sch Med, Dept Obstet & Gynecol, Baltimore, MD 21205 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Texas Dept Publ Hlth, Bur Communicable Dis Control, Austin, TX USA. Louisiana State Univ, Hlth Sci Ctr, Dept Obstet & Gynecol, New Orleans, LA USA. Maimonides Hosp, Dept Obstet & Gynecol, Brooklyn, NY 11219 USA. Univ Miami, Sch Med, Dept Obstet & Gynecol, Miami, FL 33101 USA. New York State Dept Hlth, AIDS Inst, New York, NY USA. Univ Miami, Sch Med, Dept Pediat, Miami, FL USA. Univ Calif San Diego, Dept Pediat, Div Infect Dis, La Jolla, CA 92093 USA. Brigham & Womens Hosp, Dept Obstet & Gynecol, Boston, MA 02115 USA. Albany Med Ctr, Childrens Hosp, Dept Pediat, Albany, NY USA. Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Pediat, New Brunswick, NJ 08903 USA. Univ Puerto Rico, Sch Med, Dept Obstet & Gynecol, Rio Piedras, PR 00931 USA. NIAID, Pediat Branch, Div AIDS, NIH, Bethesda, MD 20892 USA. NICHHD, Pediat Adolescent & Maternal AIDS Branch, NIH, Rockville, MD USA. HIV AIDS Bur, Div Training & Tech Assistance, Hlth Resources & Serv Adm, Rockville, MD USA. Maternal & Child Hlth Bur, Div Perinatal Syst & Womens Hlth, Hlth Resources & Serv Adm, Rockville, MD USA. US FDA, Div Antiviral Drug Prod, Rockville, MD 20857 USA. Off HIV AIDS Policy, Washington, DC USA. RP Jamieson, DJ (reprint author), CDCP, Div HIV AIDS, Natl Ctr HIV STD & TB Prevent, Mailstop E-45,1600 Clifton Rd, Atlanta, GA 30333 USA. NR 19 TC 12 Z9 13 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD JUN PY 2002 VL 99 IS 6 BP 1117 EP 1126 AR PII S0029-7844(02)01985-3 PG 10 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 558QF UT WOS:000175977500029 ER PT J AU Thompson, DJS Barron, KS Whitcup, SM Robinson, MR AF Thompson, DJS Barron, KS Whitcup, SM Robinson, MR TI The safety and efficacy of chicken type II collagen on uveitis associated with juvenile rheumatoid arthritis SO OCULAR IMMUNOLOGY AND INFLAMMATION LA English DT Article DE uveitis; juvenile rheumatoid arthritis (JRA); oral tolerance; collagen; autoimmune ID CLINICAL RESEARCH; DOUBLE-BLIND; TRIAL; SUPPRESSION; CHILDREN; RATS AB Purpose: To investigate the safety and efficacy of chicken type II collagen in treating uveitis associated with juvenile rheumatoid arthritis (JRA). Methods: A prospective dose-ranging (60 and 540 fig) pilot study of orally administered chicken type II collagen in 13 participants, aged 2-18 years, with JRA and uveitis and without prior exposure to collagens. Anterior chamber cells, flare, vitreous haze, visual acuity, and concomitant anti-inflammatory medications were the ophthalmic outcomes. Arthritis outcomes included the American College of Rheumatology (ACR) core set. Results: No serious or related adverse events were reported. Four participants (2 low dose, 2 high dose) experienced improvement in ophthalmic outcome, while two participants (1 in each group) worsened (p > 0.5). According to ACR criteria, six participants showed improvement in JRA. Conclusions: Although appearing safe, clearly demonstrating the efficacy of this treatment for JRA or uveitis remains a challenge. Based on the results from this. pilot study, a large positive treatment effect on uveitis is unlikely. C1 EMMES Corp, Rockville, MD 20850 USA. NIAID, Div Intramural Res, Bethesda, MD 20892 USA. NEI, Clin Branch, NIH, Bethesda, MD 20892 USA. RP Thompson, DJS (reprint author), EMMES Corp, 401 N Washington St,Suite 700, Rockville, MD 20850 USA. NR 23 TC 7 Z9 9 U1 1 U2 2 PU SWETS ZEITLINGER PUBLISHERS PI LISSE PA P O BOX 825, 2160 SZ LISSE, NETHERLANDS SN 0927-3948 J9 OCUL IMMUNOL INFLAMM JI Ocul. Immunol. Inflamm. PD JUN PY 2002 VL 10 IS 2 BP 83 EP 91 DI 10.1076/ocii.10.2.83.13977 PG 9 WC Ophthalmology SC Ophthalmology GA 680NN UT WOS:000182984400001 PM 12778344 ER PT J AU Cheson, BD West, PJ AF Cheson, BD West, PJ TI Clinical trials - Referral resource: Current clinical trials in non-Hodgkin's lymphoma SO ONCOLOGY-NEW YORK LA English DT Article C1 NCI, Bethesda, MD 20892 USA. RP Cheson, BD (reprint author), NCI, Bethesda, MD 20892 USA. NR 7 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 JUN PY 2002 VL 16 IS 6 BP 762 EP + PG 3 WC Oncology SC Oncology GA 565GR UT WOS:000176361900012 PM 12088298 ER PT J AU Agrawal, M Emanuel, EJ AF Agrawal, M Emanuel, EJ TI The health economics of palliative care - The Payne/Coyne/Smith article reviewed SO ONCOLOGY-NEW YORK LA English DT Editorial Material ID RANDOMIZED CONTROLLED TRIAL; CANCER-PATIENTS; TERMINALLY ILL; COST; LIFE; END C1 NIH, Dept Clin Bioeth, Bethesda, MD 20892 USA. NR 15 TC 1 Z9 1 U1 3 U2 3 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 JUN PY 2002 VL 16 IS 6 BP 808 EP + PG 2 WC Oncology SC Oncology GA 565GR UT WOS:000176361900017 ER PT J AU Canto, MT Horowitz, AM Child, WL AF Canto, MT Horowitz, AM Child, WL TI Views of oral cancer prevention and early detection: Maryland physicians SO ORAL ONCOLOGY LA English DT Article ID STATISTICS AB The purpose of this study was to obtain in-depth information on Maryland physicians' knowledge, opinions and practices about oral cancer examinations. The qualitative descriptive study used one focus group conducted in a conference facility and nine one-on-one interviews at private medical offices. A criterion-purposeful sampling was used for selection of participants. Generally, we found low awareness of, and surprise about, Maryland's high oral cancer mortality rates. Physicians were not surprised that they detect more lesions than dentists, although most physicians did not provide oral cancer examinations on a routine basis. Physicians were interested in attending continuing medical education (CME) courses on oral cancer prevention and early detection but only if worked into other CME programs on cancer. They were very interested in having hands-on training on performing an oral cancer examination. These findings will be used to implement educational interventions for Maryland physicians to help increase early detection of oral cancers. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Natl Inst Dent & Craniofacial Res, Off Sci Policy & Anal, NIH, Bethesda, MD 20892 USA. RP Canto, MT (reprint author), Natl Inst Dent & Craniofacial Res, Off Sci Policy & Anal, NIH, 45 Ctr Dr,Bldg 45,Room 3AN44, Bethesda, MD 20892 USA. NR 17 TC 12 Z9 12 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0964-1955 J9 ORAL ONCOL JI Oral Oncol. PD JUN PY 2002 VL 38 IS 4 BP 373 EP 377 AR PII S1368-8375(01)00074-4 DI 10.1016/S1368-8375(01)00074-4 PG 5 WC Oncology; Dentistry, Oral Surgery & Medicine SC Oncology; Dentistry, Oral Surgery & Medicine GA 561JA UT WOS:000176135500009 PM 12076702 ER PT J AU Abernethy, DR Ruffolo, RR AF Abernethy, DR Ruffolo, RR TI International Union of Pharmacology Nomenclature and Pharmacological Reviews SO PHARMACOLOGICAL REVIEWS LA English DT Editorial Material C1 NIA, Intramural Res Program, Baltimore, MD 21224 USA. Wyeth Ayerst Res, Collegeville, PA USA. RP Abernethy, DR (reprint author), NIA, Intramural Res Program, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0031-6997 J9 PHARMACOL REV JI Pharmacol. Rev. PD JUN PY 2002 VL 54 IS 2 BP 159 EP 159 DI 10.1124/pr.54.2.159 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 556ZE UT WOS:000175879600001 ER PT J AU Howlett, AC Barth, F Bonner, TI Cabral, G Casellas, P Devane, WA Felder, CC Herkenham, M Mackie, K Martin, BR Mechoulam, R Pertwee, RG AF Howlett, AC Barth, F Bonner, TI Cabral, G Casellas, P Devane, WA Felder, CC Herkenham, M Mackie, K Martin, BR Mechoulam, R Pertwee, RG TI International Union of Pharmacology. XXVII. Classification of cannabinoid receptors SO PHARMACOLOGICAL REVIEWS LA English DT Review ID FATTY-ACID AMIDE; ACTIVATED PROTEIN-KINASE; CENTRAL-NERVOUS-SYSTEM; HIPPOCAMPAL ACETYLCHOLINE-RELEASE; LONG-TERM POTENTIATION; NIGRA PARS RETICULATA; NITRIC-OXIDE RELEASE; EXCITATORY SYNAPTIC-TRANSMISSION; MEDIATED SIGNAL-TRANSDUCTION; RAT CEREBELLAR MEMBRANES AB Two types of cannabinoid receptor have been discovered so far, CB1 (2.1:CBD:1:CB1:), cloned in 1990, and CB2 (2.1:CBD:2:CB2:), cloned in 1993. Distinction between these receptors is based on differences in their predicted amino acid sequence, signaling mechanisms, tissue distribution, and sensitivity to certain potent agonists and antagonists that show marked selectivity for one or the other receptor type. Cannabinoid receptors CB1 and CB2 exhibit 48% amino acid sequence identity. Both receptor types are coupled through G proteins to adenylyl cyclase and mitogen-activated protein kinase. CB1 receptors are also coupled through G proteins to several types of calcium and potassium channels. These receptors exist primarily on central and peripheral neurons, one of their functions being to inhibit neurotransmitter release. Indeed, endogenous CB1 agonists probably serve as retrograde synaptic messengers. CB2 receptors are present mainly on immune cells. Such cells also express CB1 receptors, albeit to a lesser extent, with both receptor types exerting a broad spectrum of immune effects that includes modulation of cytokine release. Of several endogenous agonists for cannabinoid receptors identified thus far, the most notable are arachidonoylethanolamide, 2-arachidonoylglycerol, and 2-arachidonylglyceryl ether. It is unclear whether these eicosanoid molecules are the only, or primary, endogenous agonists. Hence, we consider it premature to rename cannabinoid receptors after an endogenous agonist as is recommended by the International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification. Although pharmacological evidence for the existence of additional types of cannabinoid receptor is emerging, other kinds of supporting evidence are still lacking. C1 Univ Aberdeen, Inst Med Sci, Dept Biomed Sci, NC IUPHAR Subcomm Cannabinoid Receptors, Aberdeen AB25 2ZD, Scotland. Sanofi Synthelabo Rech, Montpellier, France. NIMH, Funct Neuroanat Sect, Bethesda, MD 20892 USA. NIMH, Genet Lab, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Dept Microbiol & Immunol, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. Eli Lilly & Co Ltd, Lilly Res Ctr, Surrey, England. N Carolina Cent Univ, Julius L Chambers Biomed Biotechnol Res Inst, Neurosci Drug Abuse Program, Durham, NC USA. Univ Washington, Dept Anesthesiol, Seattle, WA 98195 USA. Hebrew Univ Jerusalem, Fac Med, Dept Nat Prod, Jerusalem, Israel. RP Pertwee, RG (reprint author), Univ Aberdeen, Inst Med Sci, Dept Biomed Sci, NC IUPHAR Subcomm Cannabinoid Receptors, Aberdeen AB25 2ZD, Scotland. EM rgp@aberdeen.ac.uk RI Mackie, Kenneth/B-7358-2011; Pertwee, Roger/E-1312-2011; Mackie, Ken/E-3715-2013; OI Mackie, Ken/0000-0001-8501-6199; Herkenham, Miles/0000-0003-2228-4238; Howlett, Allyn/0000-0002-2810-0164; Pertwee, Roger/0000-0003-3227-2783 NR 441 TC 1483 Z9 1550 U1 16 U2 99 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0031-6997 EI 1521-0081 J9 PHARMACOL REV JI Pharmacol. Rev. PD JUN PY 2002 VL 54 IS 2 BP 161 EP 202 DI 10.1124/pr.54.2.161 PG 42 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 556ZE UT WOS:000175879600002 PM 12037135 ER PT J AU Murphy, PM AF Murphy, PM TI International Union of Pharmacology. XXX. Update on chemokine receptor nomenclature SO PHARMACOLOGICAL REVIEWS LA English DT Review ID PROTEIN-COUPLED RECEPTOR-2; CUTTING EDGE; EXPRESSION CLONING; IDENTIFICATION; CORECEPTOR; STRL33; LIGAND; BINDS; CCR10; CXC AB An update of the International Union of Pharmacology nomenclature for chemokines is outlined, defining one new receptor type, CXCR6, and disqualifying the putative receptor, CCR11. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Murphy, PM (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Room 11N113, Bethesda, MD 20892 USA. NR 16 TC 241 Z9 256 U1 0 U2 2 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0031-6997 J9 PHARMACOL REV JI Pharmacol. Rev. PD JUN PY 2002 VL 54 IS 2 BP 227 EP 229 DI 10.1124/pr.54.2.227 PG 3 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 556ZE UT WOS:000175879600005 PM 12037138 ER PT J AU Spedding, M Bonner, TI Watson, SP AF Spedding, M Bonner, TI Watson, SP TI International Union of Pharmacology. XXXI. Recommendations for the nomenclature of multimeric G protein-coupled receptors SO PHARMACOLOGICAL REVIEWS LA English DT Review AB A receptor is defined by the International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC-IUPHAR) as a protein, or a complex of proteins, which recognizes physiologically relevant ligands that can regulate the protein to mediate cellular events (Ruffolo et al., 2000). This definition does not include associated proteins, which are not required for agonist recognition and/or receptor assembly. Thus, G proteins are not included in the nomenclature of G protein-coupled receptors (GPCRs). Similarly, proteins which modify receptor disposition, such as proteins with a PDZ domain (Sheng and Sala, 2001), and which associate with the cytosolic portion of the receptor are not included. The question arises, however, as to the way to name multimeric receptors where subunits influence receptor assembly and agonist recognition. The essential issue is whether to name the individual proteins or the association of proteins? NC-IUPHAR recommends that, where possible, the functional receptor complex be given a different name from that of the subunits. C1 IRIS, F-92578 Neuilly Sur Seine, France. NIMH, Genet Lab, Bethesda, MD 20892 USA. Univ Oxford, Dept Pharmacol, Oxford OX1 3QT, England. RP Spedding, M (reprint author), IRIS, 192 Ave Charles de Gaulle, F-92578 Neuilly Sur Seine, France. RI Watson, Stephen/Q-6292-2016 OI Watson, Stephen/0000-0002-7846-7423 NR 4 TC 5 Z9 6 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0031-6997 J9 PHARMACOL REV JI Pharmacol. Rev. PD JUN PY 2002 VL 54 IS 2 BP 231 EP 232 DI 10.1124/pr.54.2.231 PG 2 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 556ZE UT WOS:000175879600006 PM 12037139 ER PT J AU Bowery, NG Bettler, B Froestl, W Gallagher, JP Marshall, F Raiteri, M Bonner, TI Enna, SJ AF Bowery, NG Bettler, B Froestl, W Gallagher, JP Marshall, F Raiteri, M Bonner, TI Enna, SJ TI International Union of Pharmacology. XXXIII. Mammalian gamma-aminobutyric acid(B) receptors: Structure and function SO PHARMACOLOGICAL REVIEWS LA English DT Review ID LOWER ESOPHAGEAL SPHINCTER; GABA-B-RECEPTORS; RAT SPINAL-CORD; CONTINUOUS INTRATHECAL BACLOFEN; CENTRAL-NERVOUS-SYSTEM; SUBSTANTIA-GELATINOSA NEURONS; COGNITION-ENHANCING ACTIVITY; CEREBRAL CORTICAL MEMBRANES; DISCRETE TRIALS PROCEDURE; LONG-TERM POTENTIATION AB The gamma-aminobutyric acid(B) (GABA(B)) receptor was first demonstrated on presynaptic terminals where it serves as an autoreceptor and also as a heteroreceptor to influence transmitter release by suppressing neuronal Ca2+ conductance. Subsequent studies showed the presence of the receptor on postsynaptic neurones where activation produces an increase in membrane K+ conductance and associated neuronal hyperpolarization. (-)-Baclofen is a highly selective agonist for GABA(B) receptors, whereas the established GABA(A) receptor antagonists, bicuculline and picrotoxin, do not block GABA(B) receptors. The receptor is G(i)/G(o) protein-coupled with mixed effects on adenylate cyclase activity. The receptor comprises a heterodimer with similar subunits currently designated 1 and 2. These subunits are coupled via coiled-coil domains at their C termini. The evidence for splice variants is critically reviewed. Thus far, no unique pharmacological or functional properties have been assigned to either subunit or the variants. The emergence of high-affinity antagonists for GABA(B) receptors has enabled a synaptic role to be established. However, the antagonists have generally failed to establish the existence of pharmacologically distinct receptor types within the GABA(B) receptor class. The advent of GABA(B1) knockout mice has also failed to provide support for multiple receptor types. C1 Univ Birmingham, Sch Med, Dept Pharmacol, Birmingham B15 2TT, W Midlands, England. Univ Basel, Pharmactr, Basel, Switzerland. Novartis Pharmaceut, Nervous Syst Res, Basel, Switzerland. Univ Texas, Med Branch, Dept Pharmacol & Toxicol, Galveston, TX 77550 USA. Millennium Pharmaceut Inc, Granta Pk, Great Abington, England. Univ Genoa, Dept Expt Med Pharmacol & Toxicol, Genoa, Italy. NIMH, Genet Lab, Bethesda, MD 20892 USA. Univ Kansas, Sch Med, Dept Pharmacol Toxicol & Therapeut, Kansas City, KS USA. RP Bowery, NG (reprint author), Univ Birmingham, Sch Med, Dept Pharmacol, Birmingham B15 2TT, W Midlands, England. EM n.g.bowery@bham.ac.uk NR 318 TC 370 Z9 391 U1 3 U2 13 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0031-6997 EI 1521-0081 J9 PHARMACOL REV JI Pharmacol. Rev. PD JUN PY 2002 VL 54 IS 2 BP 247 EP 264 DI 10.1124/pr.54.2.247 PG 18 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 556ZE UT WOS:000175879600008 PM 12037141 ER PT J AU Shi, YB Ritchie, JWA Taylor, PM AF Shi, YB Ritchie, JWA Taylor, PM TI Complex regulation of thyroid hormone action: multiple opportunities for pharmacological intervention SO PHARMACOLOGY & THERAPEUTICS LA English DT Review DE histone acetylation; nuclear receptor; thyroid hormone transporter; nuclear hormone action; gene regulation ID AMINO-ACID-TRANSPORT; LIGAND-BINDING DOMAIN; PROTEIN DISULFIDE ISOMERASE; NUCLEAR RECEPTOR; TRANSCRIPTIONAL REPRESSION; HISTONE ACETYLATION; DNA-BINDING; XENOPUS-LAEVIS; N-COR; AMPHIBIAN METAMORPHOSIS AB The thyroid hormone (TH; 3,3',5,5'-tetra-iodothyronine and 3,3',5-triiodothyronine) regulates growth, development, and critical metabolic functions. Thyroid diseases are among the most prevalent group of metabolic disorders in the Western world. TH exert's effects through complex biological pathways, which offer a wealth of opportunities to pharmacologically intervene in TH signalling at numerous steps. These include biosynthesis, cell-specific uptake or export (involving L-type amino acid transporter, organic anion transporter, organic cation transporter, or multidrug resistance transporter), as well as nuclear targeting and actions (the latter including TH receptor binding and histone acetylation/deacetylation). Such processes represent potentially important pharmacological targets for the design of novel or improved therapies for TH disorders, obesity, and cardiovascular diseases. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NICHHD, Unit Mol Morphogenesis, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA. Prolifix Ltd, Abingdon OX14 4RY, Oxon, England. Univ Dundee, Sch Life Sci, Div Mol Physiol, Dundee DD1 5EH, Scotland. RP Shi, YB (reprint author), NICHHD, Unit Mol Morphogenesis, Lab Gene Regulat & Dev, NIH, Bldg 18T,Room 106, Bethesda, MD 20892 USA. RI Taylor, Peter/A-4667-2010 NR 166 TC 24 Z9 27 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0163-7258 J9 PHARMACOL THERAPEUT JI Pharmacol. Ther. PD JUN PY 2002 VL 94 IS 3 BP 235 EP 251 AR PII S0163-7258(02)00219-X DI 10.1016/S0163-7258(02)00219-X PG 17 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 582CV UT WOS:000177332900003 PM 12113800 ER PT J AU Rudek, MA Venitz, J Figg, WD AF Rudek, MA Venitz, J Figg, WD TI Matrix metalloproteinase inhibitors: Do they have a place in anticancer therapy? SO PHARMACOTHERAPY LA English DT Review ID CHEMICALLY-MODIFIED TETRACYCLINES; RHEUMATOID SYNOVIAL FIBROBLASTS; SQUAMOUS-CELL CARCINOMA; IN-SITU HYBRIDIZATION; NECROSIS-FACTOR-ALPHA; GELATINASE-A MMP-2; PHASE-I; MESSENGER-RNA; TISSUE INHIBITOR; BAY 12-9566 AB Matrix metalloproteinases (MMPs) are a family of enzymes involved in degradation of extracellular matrix. An imbalance between MMPs and naturally occurring MMP inhibitors may cause excess extracellular matrix destruction, allowing cancer cells to invade surrounding tissues and metastasize, and permitting angiogenesis to occur. Inhibition of certain key MMPs may prevent angiogenesis, tumor growth, invasion, and metastasis. Gelatinases MMP-2 and MMP-9 are expressed during carcinogenesis and angiogenesis. Synthetic MMP inhibitors were designed to target these enzymes and potentially prevent the tumor growth and metastases associated with cancer. C1 NCI, Mol Pharmacol Sect, Canc Therapeut Branch, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. NCI, Med Oncol Clin Res Unit, Ctr Canc Res, NIH, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmaceut, Sch Pharm, Richmond, VA 23298 USA. RP Figg, WD (reprint author), NCI, Mol Pharmacol Sect, Canc Therapeut Branch, Ctr Canc Res,NIH, Bldg 10,Room 5A01,MSC1910,9000 Rockville Pike, Bethesda, MD 20892 USA. EM wdfigg@helix.nih.gov RI Figg Sr, William/M-2411-2016 NR 161 TC 38 Z9 41 U1 0 U2 2 PU PHARMACOTHERAPY PUBLICATIONS INC PI BOSTON PA NEW ENGLAND MEDICAL CENTER, 806, 750 WASHINGTON ST, BOSTON, MA 02111 USA SN 0277-0008 J9 PHARMACOTHERAPY JI Pharmacotherapy PD JUN PY 2002 VL 22 IS 6 BP 705 EP 720 DI 10.1592/phco.22.9.705.34062 PG 16 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 560FM UT WOS:000176072100005 PM 12066962 ER PT J AU Berezhkovskii, AM Sutmann, G AF Berezhkovskii, AM Sutmann, G TI Time and length scales for diffusion in liquids SO PHYSICAL REVIEW E LA English DT Article AB The first six even moments of the displacement of a molecule in water and an atom in liquid argon are found by molecular dynamics simulations and compared with the moments predicted by diffusion theory. We find a noticeable difference between the moments higher than the second. The ratio between predicted and calculated moments approaches unity as 1/t for times larger than 10 ps. Continuous time random walk is used to explain this slow approach of the moments to their diffusion limit. C1 NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. Res Ctr Julich, Cent Inst Appl Math, D-52425 Julich, Germany. Res Ctr Julich, John von Neumann Inst Comp, D-52425 Julich, Germany. Karpov Inst Phys Chem, Moscow 103064, Russia. RP NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. EM berezh@speck.niddk.nih.gov; g.sutmann@fz-juelich.de NR 15 TC 10 Z9 10 U1 0 U2 3 PU AMER PHYSICAL SOC PI COLLEGE PK PA ONE PHYSICS ELLIPSE, COLLEGE PK, MD 20740-3844 USA SN 2470-0045 EI 2470-0053 J9 PHYS REV E JI Phys. Rev. E PD JUN PY 2002 VL 65 IS 6 AR 060201 DI 10.1103/PhysRevE.65.060201 PN 1 PG 4 WC Physics, Fluids & Plasmas; Physics, Mathematical SC Physics GA 572FH UT WOS:000176762500003 PM 12188692 ER PT J AU Toyofuku, K Valencia, JC Kushimoto, T Costin, GE Virador, VM Vieira, WD Ferrans, VJ Hearing, VJ AF Toyofuku, K Valencia, JC Kushimoto, T Costin, GE Virador, VM Vieira, WD Ferrans, VJ Hearing, VJ TI The etiology of oculocutaneous albinism (OCA) type II: The pink protein modulates the processing and transport of tyrosinase SO PIGMENT CELL RESEARCH LA English DT Article DE tyrosinase; oculocutaneous; albinism; Tyrp1; Pmel17; Dct ID EYED DILUTION GENE; ULTRASTRUCTURAL-LOCALIZATION; ENDOPLASMIC-RETICULUM; MAMMALIAN TYROSINASE; MELANOSOMAL PROTEINS; MOLECULAR-BASIS; B-16 MELANOMA; MOUSE; MELANOCYTES; EXPRESSION AB Oculocutaneous albinism (OCA) is caused by reduced or deficient melanin pigmentation in the skin, hair, and eyes. OCA has different phenotypes resulting from mutations in distinct pigmentation genes involved in melanogenesis. OCA type 2 (OCA2), the most common form of OCA, is an autosomal recessive disorder caused by mutations in the P gene, the function(s) of which is controversial. In order to elucidate the mechanism(s) involved in OCA2, our group used several antibodies specific for various melanosomal proteins (tyrosinase, Tyrp1, Dct, Pmel17 and HMB45), including a specific set of polyclonal antibodies against the p protein. We used confocal immunohistochemistry to compare the processing and distribution of those melanosomal proteins in wild type (melan-a) and in p mutant (melan-p1) melanocytes. Our results indicate that the melanin content of melan-p1 melanocytes was less than 50% that of wild type melan-a melanocytes. In contrast, the tyrosinase activities were similar in extracts of wild type and p mutant melanocytes. Confocal microscopy studies and pulse-chase analyses showed altered processing and sorting of tyrosinase, which is released from melan-p1 cells to the medium. Processing and sorting of Tyrp1 was also altered to some extent. However, Dct and Pmel17 expression and subcellular localization were similar in melan-a and in melan-p1 melanocytes. In melan-a cells, the p protein showed mainly a perinuclear pattern with some staining in the cytoplasm where some co-localization with HMB45 antibody was observed. These findings suggest that the p protein plays a major role in modulating the intracellular transport of tyrosinase and a minor role for Tyrp1, but is not critically involved in the transport of DO and Pmel17. This study provides a basis to understand the relationship of the p protein with tyrosinase function and melanin synthesis, and also provides a rational approach to unveil the consequences of P gene mutations in the pathogenesis of OCA2. C1 NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Hearing, VJ (reprint author), NCI, Cell Biol Lab, NIH, Bldg 37,Room 1B25, Bethesda, MD 20892 USA. NR 43 TC 78 Z9 79 U1 0 U2 9 PU BLACKWELL MUNKSGAARD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0893-5785 J9 PIGM CELL RES JI Pigm. Cell. Res. PD JUN PY 2002 VL 15 IS 3 BP 217 EP 224 DI 10.1034/j.1600-0749.2002.02007.x PG 8 WC Cell Biology; Dermatology SC Cell Biology; Dermatology GA 559CD UT WOS:000176006500007 PM 12028586 ER PT J AU Brandizzi, F Snapp, EL Roberts, AG Lippincott-Schwartz, J Hawes, C AF Brandizzi, F Snapp, EL Roberts, AG Lippincott-Schwartz, J Hawes, C TI Membrane protein transport between the endoplasmic reticulum and the golgi in tobacco leaves is energy dependent but cytoskeleton independent: Evidence from selective photobleaching SO PLANT CELL LA English DT Article ID GREEN FLUORESCENT PROTEIN; BREFELDIN-A; LIVING CELLS; ARABIDOPSIS-THALIANA; VESICLE TRAFFICKING; SECRETORY PATHWAY; STORAGE VACUOLES; MATRIX PROTEINS; FUSION PROTEIN; APPARATUS AB The mechanisms that control protein transport between the endoplasmic reticulum (ER) and the Golgi apparatus are poorly characterized in plants. Here, we examine in tobacco leaves the structural relationship between Golgi and ER membranes using electron microscopy and demonstrate that Golgi membranes contain elements that are in close association and/or in direct contact with the ER. We further visualized protein trafficking between the ER and the Golgi using Golgi marker proteins tagged with green fluorescent protein. Using photobleaching techniques, we showed that Golgi membrane markers constitutively cycle to and from the Golgi in an energy-dependent and N-ethylmaleimide-sensitive manner. We found that membrane protein transport toward the Golgi occurs independently of the cytoskeleton and does not require the Golgi to be motile along the surface of the ER. Brefeldin A treatment blocked forward trafficking of Golgi proteins before their redistribution into the ER. Our results indicate that in plant cells, the Golgi apparatus is a dynamic membrane system whose components continuously traffic via membrane trafficking pathways regulated by brefeldin A- and N-ethylmaleimide-sensitive machinery. C1 Oxford Brookes Univ, Res Sch Biol & Mol Sci, Oxford OX3 0BP, England. NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. Scottish Crop Res Inst, Dundee DD2 5DA, Scotland. RP Brandizzi, F (reprint author), Oxford Brookes Univ, Res Sch Biol & Mol Sci, Gipsy Lane, Oxford OX3 0BP, England. RI Roberts, Alison/G-6305-2012 NR 58 TC 219 Z9 224 U1 1 U2 13 PU AMER SOC PLANT BIOLOGISTS PI ROCKVILLE PA 15501 MONONA DRIVE, ROCKVILLE, MD 20855 USA SN 1040-4651 J9 PLANT CELL JI Plant Cell PD JUN PY 2002 VL 14 IS 6 BP 1293 EP 1309 DI 10.1105/tpc.001586 PG 17 WC Biochemistry & Molecular Biology; Plant Sciences; Cell Biology SC Biochemistry & Molecular Biology; Plant Sciences; Cell Biology GA 572BL UT WOS:000176753600009 PM 12084828 ER PT J AU Voorhees, CC Schreiber, GB Schumann, BC Biro, F Crawford, PB AF Voorhees, CC Schreiber, GB Schumann, BC Biro, F Crawford, PB TI Early predictors of daily smoking in young women: The National Heart, Lung, and Blood Institute Growth and Health Study SO PREVENTIVE MEDICINE LA English DT Article DE daily smoking; predictors; young adults; women; girls ID CIGARETTE-SMOKING; SUBSTANCE USE; ADOLESCENT SMOKING; WEIGHT CONCERNS; UNITED-STATES; STRESS; VALIDITY; SAMPLE; ONSET; BLACK AB Background. Smoking is highly prevalent in young women and little is known about early multilevel independent risk or protective factors that are predictive of daily smoking in young women. Methods. Multiple logistic regression was conducted on data from NGHS, a 10-year cohort study of Black (1,213) and White (1,166) girls recruited from three clinical centers in the United States, ages 9-10 years on entry to ages 18-19. Results. Compared with never smokers, White girls were at higher risk than Black girls of being daily smokers at ages 18-19. Early predictors of daily smoking at ages 18-19 years included lower parental education, one parent in the household, drinking alcohol at ages 11-12, higher drive for thinness at ages 11-12, lower behavioral conduct at ages 11-12, and lower stress at ages 10-11 and higher stress at ages 12-13. For both Black and White girls weight-related variables were significant. Stress, behavioral conduct, and one-parent household were also important predictors for White girls. Conclusions. There is evidence that childhood and adolescent factors are related to young adult smoking behavior. Body weight concerns as well as family, social environment, and behavioral factors are important issues in determining which girls will become daily smokers. (C) 2002 American Health Foundation and Elsevier Science (USA). C1 NHLBI, Div Epidemiol & Clin Applicat, Rockville, MD USA. Westat Corp, Rockville, MD 20850 USA. Maryland Med Res Inst, Baltimore, MD USA. Childrens Hosp, Med Ctr, Div Adolescent Med, Cincinnati, OH 45229 USA. Univ Calif Berkeley, Dept Nutr Sci, Berkeley, CA 94720 USA. RP Voorhees, CC (reprint author), Johns Hopkins Univ, Sch Med, 1830 E Monument St,Room 8036, Baltimore, MD 21205 USA. FU NHLBI NIH HHS [N0-HC-55023-26, U01-HL48941-44] NR 39 TC 47 Z9 48 U1 3 U2 5 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0091-7435 J9 PREV MED JI Prev. Med. PD JUN PY 2002 VL 34 IS 6 BP 616 EP 624 DI 10.1006/pmed.2002.1026 PG 9 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 559MH UT WOS:000176029600008 PM 12052022 ER PT J AU White, JD AF White, JD TI Cancer: current research in alternative therapies SO PRIMARY CARE LA English DT Article ID PROSTATE-CANCER; PC-SPES; SHARK CARTILAGE; VISCUM-ALBUM; DIETARY-SUPPLEMENT; CLINICAL-TRIAL; BREAST-CANCER; UNITED-STATES; PHASE I/II; FAT INTAKE AB Recent surveys demonstrate that between 9% and 91% of US patients with cancer use complementary and alternative medicine (CAM) therapies at some time after their diagnosis; however, there is a paucity of data available to indicate whether these CAM practices are efficacious and safe. A recent growth of interest in CAM also has resulted in an increase in research resources focused on this subject. The National Cancer Institute and the National Center for Complementary and Alternative Medicine are establishing programs to increase the amount and quality of CAM cancer research and to support the production of high-quality CAM cancer information. C1 NCI, Off Canc Complementary & Alternat Med, Off Deputy Director Extramural Sci, NIH, Bethesda, MD 20892 USA. RP White, JD (reprint author), NCI, Off Canc Complementary & Alternat Med, Off Deputy Director Extramural Sci, NIH, Execut Plaza N,Suite 102, Bethesda, MD 20892 USA. NR 49 TC 4 Z9 4 U1 1 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0095-4543 J9 PRIMARY CARE JI Primary Care PD JUN PY 2002 VL 29 IS 2 BP 379 EP + AR PII S0095-4543(01)00002-1 DI 10.1016/S0095-4543(01)00002-1 PG 15 WC Primary Health Care; Medicine, General & Internal SC General & Internal Medicine GA 572BM UT WOS:000176753700009 PM 12391717 ER PT J AU Miles, AP Zhang, YL Saul, A Stowers, AW AF Miles, AP Zhang, YL Saul, A Stowers, AW TI Large-scale purification and characterization of malaria vaccine candidate antigen Pvs25H for use in clinical trials SO PROTEIN EXPRESSION AND PURIFICATION LA English DT Article ID PLASMODIUM-VIVAX; ANTIBODIES; RESISTANCE; YEAST AB The budding yeast Saccharomyces cerevisiae has been used to express the recombinant protein Pvs25H, currently the only candidate transmission-blocking vaccine against Plasmodium vivax malaria. This molecule contains four epidermal growth factor-like domains and is expressed as at least two stable monomeric forms with different physicochemical properties. Pvs25H-A is apparently homogeneous and seems to have a correct disulfide bond structure. By contrast, Pvs25H-B is produced as a heterogeneous population of molecules, some of which are associated with an as yet unidentified chromophore, and it contains both internal and N-terminal cleavages. We report here a procedure for successfully separating these two forms with a process suitable for clinical production of this antigen. C1 NIAID, Malaria Vaccine Dev Unit, NIH, Rockville, MD 20852 USA. RP Miles, AP (reprint author), NIAID, Malaria Vaccine Dev Unit, NIH, Rockville, MD 20852 USA. RI Saul, Allan/I-6968-2013 OI Saul, Allan/0000-0003-0665-4091 NR 14 TC 23 Z9 25 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 1046-5928 J9 PROTEIN EXPRES PURIF JI Protein Expr. Purif. PD JUN PY 2002 VL 25 IS 1 BP 87 EP 96 DI 10.1006/prep.2001.1613 PG 10 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology GA 569GG UT WOS:000176592400012 PM 12071703 ER PT J AU Halperin, I Ma, BY Wolfson, H Nussinov, R AF Halperin, I Ma, BY Wolfson, H Nussinov, R TI Principles of docking: An overview of search algorithms and a guide to scoring functions SO PROTEINS-STRUCTURE FUNCTION AND GENETICS LA English DT Review DE docking; matching; scoring; protein-ligand recognition; flexible docking ID PROTEIN-PROTEIN RECOGNITION; MOLECULAR-DYNAMICS SIMULATION; SIDE-CHAIN FLEXIBILITY; FLEXIBLE LIGAND DOCKING; MONTE-CARLO DOCKING; SURFACE SHAPE COMPLEMENTARITY; MULTIPLE STRUCTURE ALIGNMENT; HIGH-THROUGHPUT DOCKING; GENETIC ALGORITHM; HIV-1 PROTEASE AB The docking field has come of age. The time is ripe to present the principles of docking, reviewing the current state of the field. Two reasons are largely responsible for the maturity of the computational docking area. First, the early optimism that the very presence of the "correct" native conformation within the list of predicted docked conformations signals a near solution to the docking problem, has been replaced by the stark realization of the extreme difficulty of the next scoring/ranking step. Second, in the last couple of years more realistic approaches to handling molecular flexibility in docking schemes have emerged. As in folding, these derive from concepts abstracted from statistical mechanics, namely, populations. Docking and folding are interrelated. From the purely physical standpoint, binding and folding are analogous processes, with similar underlying principles. Computationally, the tools developed for docking will be tremendously useful for folding. For large, multidomain proteins, domain docking is probably the only rational way, mimicking the hierarchical nature of protein folding. The complexity of the problem is huge. Here we divide the computational docking problem into its two separate components. As in folding, solving the docking problem involves efficient search (and matching) algorithms, which cover the relevant conformational space, and selective scoring functions, which are both efficient and effectively discriminate between native and non-native solutions. It is universally recognized that docking of drugs is immensely important. However, protein-protein docking is equally so, relating to recognition, cellular pathways, and macromolecular assemblies. Proteins function when they are bound to other molecules. Consequently, we present the review from both the computational and the biological points of view. Although large, it covers only partially the extensive body of literature, relating to small (drug) and to large protein-protein molecule docking, to rigid and to flexible. Unfortunately, when reviewing these, a major difficulty in assessing the results is the non-uniformity in the formats in which they are presented in the literature. Consequently, we further propose a way to rectify it here. C1 NCI, Lab Expt & Computat Biol, Ft Detrick, MD 21702 USA. Tel Aviv Univ, Sackler Sch Med, Sackler Inst Mol Med, Dept Human Genet & Mol Med, IL-69978 Tel Aviv, Israel. Tel Aviv Univ, Sch Comp Sci, Fac Exact Sci, IL-69978 Tel Aviv, Israel. NCI, Lab Exptl & Computat Biol, Intramural Res Support Program, SAIC, Frederick, MD 21701 USA. RP Nussinov, R (reprint author), NCI, Lab Expt & Computat Biol, Bldg 469,Rm 151, Ft Detrick, MD 21702 USA. RI Wolfson, Haim/A-1837-2011; Ma, Buyong/F-9491-2011 OI Ma, Buyong/0000-0002-7383-719X FU NCI NIH HHS [N01-CO-12400] NR 280 TC 740 Z9 754 U1 19 U2 174 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-3585 J9 PROTEINS JI Proteins PD JUN 1 PY 2002 VL 47 IS 4 BP 409 EP 443 DI 10.1002/prot.10115 PG 35 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 557MB UT WOS:000175911400001 PM 12001221 ER PT J AU Markham, MR Dougher, MJ Augustson, EM AF Markham, MR Dougher, MJ Augustson, EM TI Transfer of operant discrimination and respondent elicitation via emergent relations of compound stimuli SO PSYCHOLOGICAL RECORD LA English DT Article ID ARBITRARY MATCHING TASKS; COMPLEX STIMULI; CONDITIONAL DISCRIMINATION; EQUIVALENCE-RELATIONS; IDENTITY; SAMPLES; CHILDREN AB Two experiments investigated transfer of stimulus functions via emergent relations of compound stimuli. In Experiment 1, 4 college students were taught nine conditional relations of compound stimuli and unitary stimuli (A1B1-C1, A1B2-C3, A1B3-C2, A2B1-C3, A2B2-C2, A2B3-C1, A3B1-C2, A3132-Cl, and A3B3-C3), then tested for 18 untrained stimulus relations derived from the trained relations (e.g., A1C1-B1; B1C1-A1). Participants were then taught to sequence the A stimuli (A1 -->A2-->A3), and tested for transfer of this sequencing response to BC compounds. Two participants demonstrated transfer of the sequencing response. Two participants demonstrated transfer of the sequencing response after additional experimental phases. In Experiment 2, 5 college students were taught nine AB-C relations and then tested for 18 AC-B and BC-A relations as in Experiment 1. A skin conductance response was conditioned to A1 by pairing this stimulus with mild electric shock. Participants were then tested for transfer of this skin conductance response to B1C1 and B3C2. Three participants showed the transfer of conditioning. One participant did not demonstrate conditioning of the skin conductance response. One participant showed transfer of the skin conductance response after a supplemental conditioning phase. Initial failures to show transfer for some participants suggest that transfer of function sometimes depends upon a history of differential responding to compound stimuli. These results suggest that emergent relations involving compound stimuli and stimulus equivalence are related phenomena. C1 Florida Int Univ, Miami, FL 33199 USA. Univ New Mexico, Albuquerque, NM 87131 USA. NCI, Bethesda, MD 20892 USA. RP Markham, MR (reprint author), Dept Biol Sci, Univ Pk, Miami, FL 33199 USA. OI Markham, Michael/0000-0002-2900-0077 NR 31 TC 11 Z9 13 U1 0 U2 1 PU PSYCHOLOGICAL RECORD PI GAMBIER PA KENYON COLLEGE, GAMBIER, OH 43022 USA SN 0033-2933 J9 PSYCHOL REC JI Psychol. Rec. PD SUM PY 2002 VL 52 IS 3 BP 325 EP 350 PG 26 WC Psychology, Multidisciplinary SC Psychology GA 568FV UT WOS:000176532500006 ER PT J AU Abi-Saab, W Seibyl, JP D'Souza, DC Karper, LP Gueorgueva, R Abi-Dargham, A Wong, ML Rajhans, S Erdos, JP Heninger, GR Charney, DS Krystal, JH AF Abi-Saab, W Seibyl, JP D'Souza, DC Karper, LP Gueorgueva, R Abi-Dargham, A Wong, ML Rajhans, S Erdos, JP Heninger, GR Charney, DS Krystal, JH TI Ritanserin antagonism of m-chlorophenylpiperazine effects in neuroleptic-free schizophrenics patients: support for serotonin-2 receptor modulation of schizophrenia symptoms SO PSYCHOPHARMACOLOGY LA English DT Article DE serotonin; schizophrenia; 5-HT2 receptor (5-HT2A, 5-HT2C); m-chlorophenylpiperazine; ritanserin; psychosis ID ATYPICAL ANTIPSYCHOTIC-DRUGS; DISCRIMINATIVE STIMULUS PROPERTIES; INVERSE AGONIST ACTIVITY; META-CHLOROPHENYLPIPERAZINE; 5-HT2C RECEPTORS; BEHAVIORAL-RESPONSES; CLOZAPINE TREATMENT; MEDIATED RESPONSES; ACUTE EXACERBATION; NEGATIVE SYMPTOMS AB Rationale: Serotonin-2 (5-HT2) receptor antagonism has been hypothesized to have antipsychotic activity. However, there has been limited evidence directly linking 5-HT2 receptor antagonism to symptom control in schizophrenic patients. Objectives: In order to test this hypothesis this study evaluated the capacity of pretreatment with the 5-HT2 receptor antagonist ritanserin to attenuate the effects of the 5-HT agonist, m-chlorophenylpiperazine (mCPP). Methods: Twenty-two male inpatients who met DSM-III-R criteria for schizophrenia or schizoaffective disorder completed 4 test days during which 10 mg ritanserin or matched placebo was administered orally 50 min prior to the intravenous infusion of 0.1 mg/kg mCPP or saline. The test days were conducted in a randomized order under double-blind conditions. Results: mCPP mildly and transiently increased positive symptoms and behavioral activation but not negative symptoms, as assessed by the Brief Psychiatric Rating Scale. mCPP also raised plasma prolactin and cortisol levels. All of these effects were attenuated by ritanserin pretreatment. Conclusions: These data support a contribution of 5-HT2 receptor stimulation to symptom exacerbation in schizophrenic patients and a role for 5-HT2 receptor antagonism in the prevention of this effect. C1 Yale Univ, Sch Med, Dept Psychiat, New Haven, CT 06520 USA. VA Connecticut Healthcare Syst, Schizophrenia Biol Res Ctr, Psychiat Serv 116 A, West Haven, CT 06516 USA. Yale Univ, Sch Epidemiol & Publ Hlth, Dept Stat, New Haven, CT 06519 USA. Columbia Univ Coll Phys & Surg, Dept Psychiat, New York State Psychiat Inst, New York, NY 10032 USA. Univ Calif Los Angeles, Dept Psychiat, Los Angeles, CA USA. NIMH, Intramural Res Program, Anxiety & Mood Disorders Program, Bethesda, MD 20892 USA. RP Krystal, JH (reprint author), Yale Univ, Sch Med, Dept Psychiat, 333 Cedar St, New Haven, CT 06520 USA. RI Wong, Ma-Li/D-7903-2011 NR 80 TC 15 Z9 15 U1 1 U2 3 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0033-3158 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD JUN PY 2002 VL 162 IS 1 BP 55 EP 62 DI 10.1007/s00213-002-1057-7 PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 572NN UT WOS:000176781700010 PM 12107618 ER PT J AU Munzar, P Justinova, Z Kutkat, SW Ferre, S Goldberg, SR AF Munzar, P Justinova, Z Kutkat, SW Ferre, S Goldberg, SR TI Adenosinergic modulation of the discriminative-stimulus effects of methamphetamine in rats SO PSYCHOPHARMACOLOGY LA English DT Article DE methamphetamine; adenosine; DPCPX; DMPX; caffeine; SCH 23390; spiperone; drug discrimination; rat ID A(1) RECEPTORS; DOPAMINE D-1; C-FOS; BEHAVIORAL-RESPONSES; COCAINE WITHDRAWAL; TURNING BEHAVIOR; A(2A) RECEPTORS; CAFFEINE; AMPHETAMINE; AGONISTS AB Rationale: A(1) and A(2A) adenosine receptors are co-localized with dopamine D-1 and D-2 receptors, respectively, and their stimulation attenuates dopaminergic functioning. Objective: To test whether adenosine antagonists with different selectivities for A(1) and A(2A) receptors mimic the discriminative-stimulus effects of dopamine releaser methamphetamine. Methods: Effects of the A(1) antagonist DPCPX, the preferential A(2A) antagonist DMPX and the non-selective adenosine antagonist caffeine were evaluated in Sprague-Dawley rats trained to discriminate 1.0 mg/kg, IP, meth amphetamine from saline under a fixed-ratio 10 schedule of food presentation. Results: The A(1) antagonist DPCPX (1.0-10.0 mg/kg) failed to substitute for methamphetamine. However, 5.6 mg/kg DPCPX shifted the methamphetamine dose-response curve to the left. The A(2A) antagonist DMPX (1.8-18.0 mg/kg) produced about 70% methamphetamine-appropriate responding and the non-selective antagonist caffeine (3.0-56.0 mg/kg) about 50% methamphetamine-appropriate responding at the highest tested doses. Both DMPX (5.6 mg/kg) and caffeine (30.0 mg/kg) shifted the methamphetamine dose-response curve to the left. Methamphetamine-like effects of DMPX were blocked fully by the D-2 antagonist spiperone (0.18 mg/kg) and partially by the D-1 antagonist SCH-23390 (0.018 mg/kg). Conclusions: Antagonism at A(2A) adenosine receptors directly mimics the discriminative-stimulus effects of methamphetamine through the interaction with dopamine receptors. Antagonism at A(1) adenosine receptors potentiates effects of lower methamphetamine doses and thus plays a rather indirect, modulatory role. C1 Natl Inst Drug Abuse, Intramural Res Program, Behav Neurosci Branch, Preclin Pharmacol Sect, Baltimore, MD 21224 USA. RP Munzar, P (reprint author), ALEXZA Mol Delivery Corp, 1001 E Meadow Circle, Palo Alto, CA 94303 USA. RI Justinova, Zuzana/A-9109-2011; Ferre, Sergi/K-6115-2014 OI Justinova, Zuzana/0000-0001-5793-7484; Ferre, Sergi/0000-0002-1747-1779 NR 40 TC 21 Z9 21 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0033-3158 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD JUN PY 2002 VL 161 IS 4 BP 348 EP 355 DI 10.1007/s00213-002-1075-5 PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 568YV UT WOS:000176573300003 PM 12073161 ER PT J AU Fantegrossi, WE Ullrich, T Rice, KC Woods, JH Winger, G AF Fantegrossi, WE Ullrich, T Rice, KC Woods, JH Winger, G TI 3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") and its stereoisomers as reinforcers in rhesus monkeys: serotonergic involvement SO PSYCHOPHARMACOLOGY LA English DT Article DE MDMA; self-administration; serotonin antagonist; rhesus monkey; amphetamine ID NONHUMAN-PRIMATES; OPTICAL ISOMERS; SELF-INJECTION; COCAINE; BRAIN; RATS; DISCRIMINATION; MICRODIALYSIS; STIMULATION; ANTAGONISTS AB Rationale: The reinforcing effects of MDMA and its enantiomers have not been extensively characterized in laboratory animals. Objectives: To investigate whether MDMA and its stereoisomers would be self-administered intravenously by rhesus monkeys (Macaca mulatta), and to assess the effects of serotonin, receptor antagonists on MDMA-maintained responding. Methods: Four adult male rhesus monkeys were maintained on a fixed ratio 10, time-out 60-s schedule for 0.01 mg/kg cocaine or saline injections. Racemic MDMA and its stereoisomers, and racemic methamphetamine were periodically substituted for cocaine or saline. In subsequent antagonist experiments, five adult rhesus monkeys (three male, two female) were maintained on a multiple dose fixed ratio 30, time-out 45-s schedule for cocaine or saline injections. Racemic MDMA and its enantiomers were periodically substituted for cocaine or saline, with or without a pre-session injection of the serotonin(2) receptor antagonists ketanserin or MDL100907. Results: In the initial self-administration experiments, MDMA and its stereoisomers generated "inverted U"-shaped self-administration curves across the dose range tested. Racemic MDMA doses between 0.01 and 0.1 mg/kg per injection, S(+)-MDMA doses between 0.003 and 0.1 mg/kg per injection, and R(-)-MDMA doses between 0.01 and 0.1 mg/kg per injection engendered more responding than saline; however, no dose of any form of MDMA maintained as much behavior as cocaine or methamphetamine. In subsequent antagonist experiments, pretreatments with 0.1 or 0.3 mg/kg ketanserin or MDL100907 attenuated responding for S(+)-MDMA, and completely abolished responding for R(-)-MDMA, but did not affect cocaine-maintained behavior. Coticlusions: MDMA and its stereoisomers serve as reinforcers in rhesus monkeys. We suggest that stimulation of 5-HT2A receptors is integral to the reinforcing effects of MDMA. C1 Univ Michigan, Dept Psychol, Biopsychol Program, Ann Arbor, MI 48109 USA. Univ Michigan, Dept Pharmacol, Med Sch, Ann Arbor, MI 48109 USA. NIDDK, Lab Med Chem, NIH, Bethesda, MD 20892 USA. RP Woods, JH (reprint author), Univ Michigan, Dept Psychol, Biopsychol Program, Ann Arbor, MI 48109 USA. FU NIDA NIH HHS [DA05923, DA09161] NR 42 TC 95 Z9 95 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0033-3158 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD JUN PY 2002 VL 161 IS 4 BP 356 EP 364 DI 10.1007/s00213-002-1021-6 PG 9 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 568YV UT WOS:000176573300004 PM 12073162 ER PT J AU Highfield, DA Mead, AN Grimm, JW Rocha, BA Shaham, Y AF Highfield, DA Mead, AN Grimm, JW Rocha, BA Shaham, Y TI Reinstatement of cocaine seeking in 129X1/SvJ mice: effects of cocaine priming, cocaine cues and food deprivation SO PSYCHOPHARMACOLOGY LA English DT Article DE conditioned drug cues; drug self-administration; extinction; reinstatement; relapse; stress ID CONDITIONED PLACE PREFERENCE; EXTRACELLULAR DOPAMINE LEVELS; TRANSPORTER KNOCKOUT MICE; STRESS-INDUCED RELAPSE; NUCLEUS-ACCUMBENS; LOCOMOTOR-ACTIVITY; RECEPTOR AGONISTS; DRUG-SEEKING; 129/J MICE; BEHAVIOR AB Rationale and objectives: The mechanisms underlying relapse to cocaine seeking induced by exposure to priming cocaine injections, cues associated with cocaine self-administration and environmental stressors have been studied in rats. Here we describe a reinstatement method for studying relapse to cocaine seeking in mice, a suitable species for studying the effect of genetic manipulations such as gene knockout or gene overexpression on compulsive drug use. Methods: Male mice of the 129X1/SvJ strain were trained for 14-16 days to self-administer cocaine (0.75 mg/kg/infusion; 4 h/day; fixed-ratio-1 schedule of reinforcement; infusions were paired with a light-tone compound cue). Next, the lever-pressing behavior was extinguished by removing the cocaine syringes in the presence (Exps. 1 and 3) or absence (Exp. 2) of the cocaine cue. Subsequently, tests for reinstatement were conducted after exposure to priming injections of cocaine (0, 1.5, 3.0 and 6.0 mg/kg, IV; Exp. 1), response-contingent presentations of the cocaine-associated cue (Exp. 2), or food deprivation stress (I and 22 h; Exp. 3). Results: The effect of cocaine priming on reinstatement was modest and was only observed at the highest dose tested. On the other hand, reinstatement of cocaine seeking was observed following exposure to the cocaine-associated cue and food deprivation stress. Conclusions: The present data suggest that factors contributing to relapse to drugs can be studied in the reinstatement model using the common 129X1/SvJ mouse inbred strain. C1 IRP NIDA NIH, Behav Neurosci Branch, Baltimore, MD 21224 USA. RP Shaham, Y (reprint author), IRP NIDA NIH, Behav Neurosci Branch, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. EM yshaham@intra.nida.nih.gov RI shaham, yavin/G-1306-2014 NR 63 TC 44 Z9 45 U1 3 U2 5 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0033-3158 EI 1432-2072 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD JUN PY 2002 VL 161 IS 4 BP 417 EP 424 DI 10.1007/s00213-002-1047-9 PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 568YV UT WOS:000176573300012 PM 12073170 ER PT J AU Kennel, SJ Mirzadeh, S Eckelman, WC Waldmann, TA Garmestani, K Yordanov, AT Stabin, MG Brechbiel, MW AF Kennel, SJ Mirzadeh, S Eckelman, WC Waldmann, TA Garmestani, K Yordanov, AT Stabin, MG Brechbiel, MW TI Vascular-targeted radioimmunotherapy with the alpha-particle emitter At-211 SO RADIATION RESEARCH LA English DT Article ID LONG-TERM SURVIVAL; HUMANIZED ANTI-TAC; MONOCLONAL-ANTIBODY; THERAPEUTIC EFFICACY; HISTOLOGIC ANALYSIS; TUMOR VASCULATURE; BLOOD-VESSELS; SOLID TUMORS; MICE; LUNG AB Astatine-211, an alpha-particle emitter, was employed in a model system for vascular-targeted radioimmunotherapy of small tumours in mouse lung to compare its performance relative to other radioisotopes in the same system. Astatine-211 was coupled to the lung blood vessel-targeting monoclonal antibody 201B with N-succinimidyl N-(4-[(211) At]astatophenethyl) succinamate linker. Biodistribution data showed that the conjugate delivered At-211 to the lung (260-418% ID/g), where it remained with a biological half-time of about 30 h. BALB/c mice bearing about 100 lung tumor colonies of EMT-6 cells, each about 2000 cells in size, were treated with At-211-labeled monoclonal antibody 201B. The administered activity of 185 kBq per animal extended the life span of treated mice over untreated controls. Injections of 370 kBq, corresponding to an absorbed dose of 25-40 Gy, were necessary to eradicate all of the lung tumors. Mice receiving 740 kBq of At-211-labeled monoclonal antibody 201B developed pulmonary fibrosis 3-4 months after treatment, as did mice treated with 3700 kBq of the alpha-particle emitter Bi-213-labeled monoclonal antibody 201B in previous work. Animals that were injected with At-211 bound to untargeted IgG or to glycine, as control agents, also demonstrated therapeutic effects relative to untreated controls. Control groups that received untargeted At-211 required about twice as much administered activity for effective therapy as did groups with lung-targeted radioisotope. These results were not consistent with radioisotope biodistribution and dosimetry calculations that indicated that lung-targeted At-211 should be at least 10-fold more efficient for lung colony therapy than At-211 bound to nontargeting controls. The data showed that At-211 is useful for vascular-targeted radioimmunotherapy because lung tumor colonies were eradicated in the mice. Work in this model system demonstrates that vascular targeting of alpha-particle emitters is an efficient therapy for small perivascular tumors and may be applicable to human disease when specific targeting agents are identified. (C) 2002 by Radiation Research Society. C1 Oak Ridge Natl Lab, Div Life Sci, Oak Ridge, TN 37831 USA. NCI, Metab Branch, Bethesda, MD USA. NCI, Radiat Oncol Branch, Ctr Canc Res, Bethesda, MD USA. Vanderbilt Univ, Nashville, TN USA. RP Kennel, SJ (reprint author), Oak Ridge Natl Lab, Div Life Sci, Bldg 4500S,Rm F150,MS 6101, Oak Ridge, TN 37831 USA. NR 43 TC 34 Z9 36 U1 3 U2 5 PU RADIATION RESEARCH SOC PI OAK BROOK PA 820 JORIE BOULEVARD, OAK BROOK, IL 60523 USA SN 0033-7587 J9 RADIAT RES JI Radiat. Res. PD JUN PY 2002 VL 157 IS 6 BP 633 EP 641 DI 10.1667/0033-7587(2002)157[0633:VTRWTA]2.0.CO;2 PG 9 WC Biology; Biophysics; Radiology, Nuclear Medicine & Medical Imaging SC Life Sciences & Biomedicine - Other Topics; Biophysics; Radiology, Nuclear Medicine & Medical Imaging GA 558PH UT WOS:000175975400004 PM 12005541 ER PT J AU Carr, ZA Kleinerman, RA Stovall, M Weinstock, RM Griem, ML Land, CE AF Carr, ZA Kleinerman, RA Stovall, M Weinstock, RM Griem, ML Land, CE TI Malignant neoplasms after radiation therapy for peptic ulcer SO RADIATION RESEARCH LA English DT Article ID GASTRIC-CANCER; PARTIAL GASTRECTOMY; DUODENAL-ULCER; HELICOBACTER-PYLORI; STOMACH-CANCER; RISK; MORTALITY; SMOKING; RADIOTHERAPY; ALCOHOL AB Most information on radiation-related cancer risk comes from the Life Span Study (LSS) of the Japanese atomic bomb survivors. Stomach cancer mortality rates are much higher in Japan than in the U.S., making the applicability of LSS findings to the U.S. population uncertain. A unique cohort of U.S. patients who were irradiated for peptic ulcer to control gastric secretion provides a different perspective on risk. Cancer mortality data were analyzed and relative risks estimated for 3719 subjects treated by radiotherapy (mean stomach dose 14.8 Gy) and/or by surgery and medication during the period 1936-1965 and followed through 1997 (average 25 years). Compared to the U.S. rates, stomach cancer mortality was significantly increased for irradiated and nonirradiated patients (observed/expected = 3.20 and 1.52, respectively). We observed strong evidence of exposure-related excess mortality from cancer of the stomach (RR 2.6, 95% Cl 1.3, 5.1), pancreas (RR 2.7, 95% Cl 1.5, 5.1), and lung (RR 1.5, 95% Cl 1.1, 2.1), with commensurate radiation dose responses in analyses that included nonexposed patients. However, the dose responses for these cancers were not significant when restricted to exposed patients. Our excess relative risk per gray estimate of 0.20 at doses less than or equal to 10 Gy (95% Cl 0, 0.73) is consistent with the estimate of 0.24 (95% Cl 0.10, 0.40) obtained from the LSS study with the linear model. (C) 2002 by Radiation Research Society. C1 NCI, REB, DCEG, NIH, Rockville, MD 20852 USA. Univ Texas, MD Anderson Canc Ctr, Dept Radiat Phys, Houston, TX 77030 USA. Univ Chicago, Med Ctr, Dept Radiat & Cellular Oncol, Chicago, IL 60637 USA. RP Carr, ZA (reprint author), NCI, REB, DCEG, NIH, 6120 Execut Blvd,Execut Plaza S,Suite 7091, Rockville, MD 20852 USA. EM abylkasz@mail.nih.gov OI Kleinerman, Ruth/0000-0001-7415-2478 NR 42 TC 46 Z9 50 U1 0 U2 2 PU RADIATION RESEARCH SOC PI LAWRENCE PA 810 E TENTH STREET, LAWRENCE, KS 66044 USA SN 0033-7587 J9 RADIAT RES JI Radiat. Res. PD JUN PY 2002 VL 157 IS 6 BP 668 EP 677 DI 10.1667/0033-7587(2002)157[0668:MNARTF]2.0.CO;2 PG 10 WC Biology; Biophysics; Radiology, Nuclear Medicine & Medical Imaging SC Life Sciences & Biomedicine - Other Topics; Biophysics; Radiology, Nuclear Medicine & Medical Imaging GA 558PH UT WOS:000175975400009 PM 12005546 ER PT J AU Grigoryan, AM Hostetter, G Kallioniemi, O Dougherty, ER AF Grigoryan, AM Hostetter, G Kallioniemi, O Dougherty, ER TI Simulation toolbox for 3D-FISH spot-counting algorithms SO REAL-TIME IMAGING LA English DT Article ID FLUORESCENCE INSITU HYBRIDIZATION; INTERPHASE CELL-NUCLEI; FISH AB Fluorescent in situ hybridization (FISH) is a molecular diagnostic technique in which a fluorescent labeled probe hybridizes to a target nucleotide sequence of DNA. Upon excitation, each chromosome containing the target sequence produces a fluorescent signal (spot). New technology provides a stack of images on multiple focal planes throughout a tissue sample. Multiple-focal-plane imaging helps to overcome the biases and imprecision inherent in single-focal-plane methods. New algorithms are being proposed to take advantage of the multiple focal planes. This paper presents a simulation toolbox that generates synthetic spots, both signal and noise, with which to evaluate algorithm performance. By using the toolbox, preset algorithm parameters can be tuned or optimized. Two critical issues for any algorithm are the sampling rate, which is the number of focal planes used, and the parameters for the filter that determines which detected spots are signal and which are noise. One would like to use a minimal number of focal planes for reasons of both time and cost, and one would like to have a noise filter that optimally balances the passing and non-passing of signal and noise spots. As demonstrated, it is possible to go further and calibrate the noise filter so that the expected number of counted spots, both signal and noise, is approximately equal to the actual number of signal spots. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Univ Texas, Dept Elect Engn, San Antonio, TX 78285 USA. NHGRI, NIH, Bethesda, MD 20892 USA. Texas A&M Univ, Dept Elect Engn, College Stn, TX 77843 USA. RP Grigoryan, AM (reprint author), Univ Texas, Dept Elect Engn, San Antonio, TX 78285 USA. RI Kallioniemi, Olli/H-5111-2011; Kallioniemi, Olli/H-4738-2012 OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332 NR 11 TC 11 Z9 11 U1 0 U2 2 PU ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1077-2014 J9 REAL-TIME IMAGING JI Real-Time Imaging PD JUN PY 2002 VL 8 IS 3 BP 203 EP 212 DI 10.1006/rtim.2001.0280 PG 10 WC Computer Science, Artificial Intelligence; Computer Science, Software Engineering; Computer Science, Theory & Methods SC Computer Science GA 591KJ UT WOS:000177878600004 ER PT J AU Yamamoto, H Teramoto, H Uetani, K Igawa, K Shimizu, E AF Yamamoto, H Teramoto, H Uetani, K Igawa, K Shimizu, E TI Cyclic stretch upregulates interleukin-8 and transforming growth factor-beta 1 production through a protein kinase C-dependent pathway in alveolar epithelial cells SO RESPIROLOGY LA English DT Article DE alveolar epithelial cells; cyclic stretch; interleukin-8; protein kinase C; transforming; growth factor-beta 1; ventilator-induced lung injury ID FACTOR-BETA; EXTRACELLULAR-MATRIX; EXPRESSION; LUNG; FIBROBLASTS; CYTOKINES; INCREASE; RELEASE; FIBRONECTIN; MACROPHAGES AB Objective: Positive-pressure mechanical ventilation can injure the lung, causing oedema and alveolar inflammation, which is termed 'ventilator-induced lung injury' (VILI). We postulated that cyclic stretch upregulates the release of cytokines, which may cause lung damage, and explored which cytokines were released after cyclic stretch in type II alveolar epithelial cells (A549). Methodology: To test this hypothesis, A549 cells were cultured on a silicoelastic membrane and interleukin (IL)-1beta, IL-8, granulocyte-macrophage colony stimulating factor, activin, transforming growth factor (TGF)-beta1, insulin-like growth factor-2 and tumour necrosis factor-alpha mRNA and protein were assessed after stimulation of the cells by cyclic stretch. Results: Cyclic stretch induced activation of protein kinase C and resulted in the release of IL-8 and TGF-beta1 from A549 cells. Conclusions: The release of IL-8 and TGF-beta1 from alveolar epithelial cells may be a contributing factor in alveolitis associated with VILI. C1 Tottori Univ, Fac Med, Dept Internal Med 3, Yonago, Tottori 683, Japan. RP Teramoto, H (reprint author), NIDR, Oral & Pharyngeal Canc Branch, NIH, 9000 Rockville Pike,Bldg 30,Room 211, Bethesda, MD 20892 USA. NR 22 TC 42 Z9 46 U1 0 U2 0 PU BLACKWELL PUBLISHING ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 1323-7799 J9 RESPIROLOGY JI Respirology PD JUN PY 2002 VL 7 IS 2 BP 103 EP 109 DI 10.1046/j.1440-1843.2002.00377.x PG 7 WC Respiratory System SC Respiratory System GA 592HJ UT WOS:000177931100003 PM 11985731 ER PT J AU Kohn, MC AF Kohn, MC TI Use of sensitivity analysis to assess reliability of metabolic and physiological models SO RISK ANALYSIS LA English DT Article ID PHARMACOKINETIC MODEL; CONTROL COEFFICIENTS; BIOCHEMICAL SYSTEMS; FLUX; INHALATION AB Because ethical considerations often preclude directly determining the human health effects of treatments or interventions by experimentation. such effects are estimated by extrapolating reactions predicted from animal experiments. Under such conditions, it must be demonstrated that the reliability of the extrapolated predictions is not excessively affected by inherent data limitations and other components of model specification. This is especially true of high-level models composed of ad hoe algebraic equations whose parameters do not correspond to specific physical properties or processes. Models based on independent experimental data restricting the numerical space of parameters that do represent actual physical properties can be represented at a more detailed level. Sensitivities of the computed trajectories to parameter variations permit more detailed attribution of uncertainties in the predictions to these low-level properties, S-systems, in which parameters are estimated empirically, and physiological models, whose parameters can be estimated accurately from independent data, are used to illustrate the applicability of trajectory sensitivity analysis to lower-level models. C1 NIEHS, Lab Computat Biol & Risk Anal, Res Triangle Pk, NC 27709 USA. RP Kohn, MC (reprint author), NIEHS, Lab Computat Biol & Risk Anal, POB 12233, Res Triangle Pk, NC 27709 USA. NR 29 TC 5 Z9 6 U1 0 U2 1 PU BLACKWELL PUBLISHERS PI MALDEN PA 350 MAIN STREET, STE 6, MALDEN, MA 02148 USA SN 0272-4332 J9 RISK ANAL JI Risk Anal. PD JUN PY 2002 VL 22 IS 3 BP 623 EP 631 DI 10.1111/0272-4332.00042 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 568NE UT WOS:000176547800022 PM 12088237 ER PT J AU Cote, CA Greer, CL Peculis, BA AF Cote, CA Greer, CL Peculis, BA TI Dynamic conformational model for the role of ITS2 in pre-rRNA processing in yeast SO RNA-A PUBLICATION OF THE RNA SOCIETY LA English DT Article DE internal transcribed spacer; ITS2; mutagenesis; pre-rRNA; ribosome; RNA structure; rRNA processing; yeast ID INTERNAL TRANSCRIBED SPACER-2; PRECURSOR RIBOSOMAL-RNA; SACCHAROMYCES-CEREVISIAE; SECONDARY STRUCTURES; FUNCTIONAL-ANALYSIS; ITS2-PROXIMAL STEM; SEQUENCE-ANALYSIS; POLYMERASE-I; 5.8S; DNA AB Maturation of the large subunit rRNAs includes a series of cleavages that result in removal of the internal transcribed spacer (ITS2) that separates mature 5.8S and 25/28S rRNAs. Previous work demonstrated that formation of higher order secondary structure within the assembling pre-ribosomal particle is a prerequisite for accurate and efficient pre-rRNA processing. To date, it is not clear which specific sequences or secondary structures are required for processing. Two alternative secondary structure models exist for Saccharomyces cerevisiae ITS2. Chemical and enzymatic structure probing and phylogenetic comparisons resulted in one structure (Yeh & Lee, J Mol Biol, 1990, 211:699-712) referred to here as the "hairpin model." More recently, an alternate folded structure was proposed (Joseph et al., Nucleic Acids Res, 1999, 27:4533-4540), called here the "ring model." We have used a functional genetic assay to examine the potential significance of these predicted structures in processing. Our data indicate that elements of both structural models are important in efficient processing. Mutations that prevent formation of ring-specific structures completely blocked production of mature 25S rRNA, whereas those that primarily disrupt hairpin elements resulted in reduced levels of mature product. Based on these results, we propose a dynamic conformational model for the role of ITS2 in processing: Initial formation of the ring structure may be required for essential, early events in processing complex assembly and may be followed by an induced transition to the hairpin structure that facilitates subsequent processing events. In this model, yeast ITS2 elements may provide in cis certain of the functions proposed for vertebrate U8 snoRNA acting In trans. C1 NIDDK, NIH, Genet & Biochem Branch, Bethesda, MD 20892 USA. Univ Calif Irvine, Dept Biol Chem, Irvine, CA 92697 USA. RP Peculis, BA (reprint author), NIDDK, NIH, Genet & Biochem Branch, 10 Ctr Dr,Room 8N322, Bethesda, MD 20892 USA. FU NIGMS NIH HHS [GM35955] NR 28 TC 71 Z9 72 U1 0 U2 3 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH ST, NEW YORK, NY 10011-4221 USA SN 1355-8382 J9 RNA JI RNA-Publ. RNA Soc. PD JUN PY 2002 VL 8 IS 6 BP 786 EP 797 DI 10.1017/S1355838202023063 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 563VF UT WOS:000176277100008 PM 12088151 ER PT J AU Prinz, W AF Prinz, W TI Cholesterol trafficking in the secretory and endocytic systems SO SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY LA English DT Review DE sterol; transport; lipid sorting; NPC1; NPC2 ID NIEMANN-PICK C1; STEROL-SENSING DOMAIN; BINDING CASSETTE TRANSPORTER-1; PLASMA-MEMBRANE; ENDOPLASMIC-RETICULUM; INTRACELLULAR-TRANSPORT; TANGIER-DISEASE; GOLGI-APPARATUS; BREFELDIN-A; VESICULAR TRAFFICKING AB Cells maintain a cholesterol gradient across the secretory system, with the lowest concentrations in the endoplasmic reticulum. (ER) and the highest in the plasma membrane (PM). Cholesterol is also heterogeneously distributed in the endocytic pathway. Little is known about how this heterogeneous distribution of cholesterol is maintained despite continuous vesicular traffic between organelles. Both the modulation of the cholesterol content of transport vesicles and the non-vesicular transport of cholesterol between organelles are likely to contribute. This review summarizes what is known about the pathways and mechanisms of intracellular sterol trafficking. C1 NIDDKD, Lab Cell Biochem & Biol, NIH, Bethesda, MD 20892 USA. RP Prinz, W (reprint author), NIDDKD, Lab Cell Biochem & Biol, NIH, Bethesda, MD 20892 USA. NR 86 TC 39 Z9 39 U1 0 U2 1 PU ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1084-9521 J9 SEMIN CELL DEV BIOL JI Semin. Cell Dev. Biol. PD JUN PY 2002 VL 13 IS 3 BP 197 EP 203 DI 10.1016/S1084-9521(02)00048-4 PG 7 WC Cell Biology; Developmental Biology SC Cell Biology; Developmental Biology GA 593DM UT WOS:000177976000006 PM 12137740 ER PT J AU Tan, AR Swain, SM AF Tan, AR Swain, SM TI Review of flavopiridol, a cyclin-dependent kinase inhibitor, as breast cancer therapy SO SEMINARS IN ONCOLOGY LA English DT Review ID CHRONIC LYMPHOCYTIC-LEUKEMIA; CELL LUNG-CANCER; ACTIVITY IN-VIVO; CARCINOMA-CELLS; DRUG FLAVOPIRIDOL; APOPTOSIS; ARREST; EXPRESSION; INDUCTION; GROWTH C1 NCI, Canc Therapeut Branch, Canc Res Ctr, Bethesda, MD 20899 USA. RP Swain, SM (reprint author), NCI, Canc Therapeut Branch, Canc Res Ctr, 8901 Wisconsin Ave,Bldg 8,Room 5101, Bethesda, MD 20899 USA. OI Swain, Sandra/0000-0002-1320-3830 NR 61 TC 20 Z9 21 U1 0 U2 4 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 JUN PY 2002 VL 29 IS 3 SU 11 BP 77 EP 85 DI 10.1053/sonc.2002.34059 PG 9 WC Oncology SC Oncology GA 578AT UT WOS:000177095800011 PM 12138401 ER PT J AU Schilsky, RL Taube, SE AF Schilsky, RL Taube, SE TI Introduction: Tumor markers as clinical cancer tests - Are we there yet? SO SEMINARS IN ONCOLOGY LA English DT Editorial Material C1 Univ Chicago, Chicago, IL 60637 USA. NCI, Bethesda, MD 20892 USA. RP Schilsky, RL (reprint author), Univ Chicago, Chicago, IL 60637 USA. NR 0 TC 27 Z9 28 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 0093-7754 J9 SEMIN ONCOL JI Semin. Oncol. PD JUN PY 2002 VL 29 IS 3 BP 211 EP 212 DI 10.1053/sonc.2002.32890 PG 2 WC Oncology SC Oncology GA 561RM UT WOS:000176156500001 PM 12063673 ER PT J AU Hammond, MEH Taube, SE AF Hammond, MEH Taube, SE TI Issues and barriers to development of clinically useful tumor markers: A development pathway proposal SO SEMINARS IN ONCOLOGY LA English DT Review ID BREAST-CANCER PATIENTS; ACTIVATOR INHIBITOR TYPE-1; MICROSATELLITE INSTABILITY; PLASMINOGEN-ACTIVATOR; PROGNOSTIC FACTORS; ADJUVANT CHEMOTHERAPY; COLORECTAL-CANCER; MONOCLONAL-ANTIBODIES; ESTROGEN-RECEPTORS; DIAGNOSTIC-TESTS C1 LDS Hosp, Dept Pathol, Salt Lake City, UT 84143 USA. Natl Canc Inst, Div Canc Treatment & Diagnosis, Canc Diagnosis Program, PACCT, Bethesda, MD USA. RP Hammond, MEH (reprint author), LDS Hosp, Dept Pathol, 8th Ave & C St, Salt Lake City, UT 84143 USA. NR 54 TC 45 Z9 49 U1 1 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0093-7754 J9 SEMIN ONCOL JI Semin. Oncol. PD JUN PY 2002 VL 29 IS 3 BP 213 EP 221 DI 10.1053/sonc.2002.32896 PG 9 WC Oncology SC Oncology GA 561RM UT WOS:000176156500002 PM 12063674 ER PT J AU Rosenwald, A Staudt, LM AF Rosenwald, A Staudt, LM TI Clinical translation of gene expression profiling in lymphomas and Leukemias SO SEMINARS IN ONCOLOGY LA English DT Review ID CHRONIC LYMPHOCYTIC-LEUKEMIA; LARGE-CELL LYMPHOMA; CLASSIFICATION; MUTATION; CANCER; NEOPLASMS C1 NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. Metab Branch, Ctr Canc Res, NCI, Bethesda, MD USA. RP Staudt, LM (reprint author), NCI, Metab Branch, NIH, Bldg 10 Rm 4N114 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 22 TC 36 Z9 37 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 0093-7754 J9 SEMIN ONCOL JI Semin. Oncol. PD JUN PY 2002 VL 29 IS 3 BP 258 EP 263 DI 10.1053/sonc.2002.32901 PG 6 WC Oncology SC Oncology GA 561RM UT WOS:000176156500006 PM 12063678 ER PT J AU Shill, H Stacy, M AF Shill, H Stacy, M TI Respiratory complications of Parkinson's disease SO SEMINARS IN RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Review DE Parkinson's disease; dopaminergic; levodopa; chest wall rigidity; pulmonary complications ID PULMONARY-FUNCTION TESTS; PLEUROPULMONARY DISEASE; DYSFUNCTION; INVOLVEMENT; THERAPY; BROMOCRIPTINE; HYPOPHONIA; LEVODOPA; FAILURE AB Patients with Parkinson's disease are at risk for pulmonary complications as a consequence of both the underlying disease pathology and the side effects of medication. Degeneration of the substantia nigra and subsequent loss of dopaminergic neurons may produce changes in ventilatory parameters. Upper airway obstruction and chest wall restriction are both common, and both may respond to levodopa. However, therapy for Parkinson's may also contribute to pulmonary morbidity. Overtreatment with levodopa causes respiratory dyskinesia that may be difficult to differentiate from complications of the disease itself. Therapy with ergot derivatives may cause pleuropulmonary fibrosis. Pneumonia resulting from the respiratory complications remains a significant cause of morbidity and mortality in Parkinson's disease. C1 Barrow Neurol Inst, Muhammad Ali Parkinson Res Ctr, Phoenix, AZ 85013 USA. NINDS, Human Motor Control Sect, NIH, Bethesda, MD 20892 USA. RP Stacy, M (reprint author), Barrow Neurol Inst, Muhammad Ali Parkinson Res Ctr, 500 W Thomas,Suite 700, Phoenix, AZ 85013 USA. NR 43 TC 10 Z9 11 U1 0 U2 0 PU THIEME MEDICAL PUBL INC PI NEW YORK PA 333 SEVENTH AVE, NEW YORK, NY 10001 USA SN 1069-3424 J9 SEM RESP CRIT CARE M JI Semin. Respir. Crit. Care Med. PD JUN PY 2002 VL 23 IS 3 BP 261 EP 265 DI 10.1055/s-2002-33034 PG 5 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 574KL UT WOS:000176888400009 PM 16088618 ER PT J AU Schaaf, MJM Cidlowski, JA AF Schaaf, MJM Cidlowski, JA TI AUUUA motifs in the 3 ' UTR of human glucocorticoid receptor alpha and beta mRNA destabilize mRNA and decrease receptor protein expression SO STEROIDS LA English DT Article DE AUUUA motif; beta-isoform; glucocorticoid receptor; RNA stability; steroid ID AU-RICH ELEMENTS; MESSENGER-RNA DEGRADATION; 3'-UNTRANSLATED REGION; GENE-EXPRESSION; IMMUNE FUNCTION; C-FOS; ISOFORM; DNA; MECHANISMS; RESISTANCE AB An association between a gene polymorphism of the human glucocorticoid receptor (hGR) gene and rheumatoid arthritis has recently been suggested. This polymorphism contains an A to G mutation in the 3'UTR of exon 9beta, which encodes the 3'UTR of the mRNA of the hGRbeta isoform. The hGRbeta isoform can act as a dominant negative inhibitor of hGRalpha, and therefore may contribute to glucocorticoid resistance. The A to G mutation is located in an AUUUA motif, which is known to destabilize mRNA. In the present study, the importance of the mutation in this AUUUA motif was further characterized and mutations in other AUUUA motifs in the 3'UTR of hGRbeta and hGRalpha mRNA were studied. hGRbeta and hGRalpha expression vectors, carrying mutations in one AUUUA motif or all AUUUA motifs were transiently transfected into COS-1 cells. Each transfected vector was analyzed for the mRNA expression level, the mRNA turnover rate and the protein expression level. The naturally occurring mutation in the 3'UTR of hGRbeta mRNA increased mRNA stability and protein expression. Mutation of two other AUUUA motifs in the 3'UTR of hGRbeta, or mutation of all four AUUUA motifs resulted in a similar effect. Mutation of the most 5' AUUUA motif did not alter hGRbeta mRNA expression or mRNA stability. Mutation of all 10 AUUUA motifs in the 3'UTR of hGRalpha mRNA increased hGRalpha mRNA expression and mRNA stability as well as expression of the receptor protein level. Thus, the naturally occurring mutation in an AUUUA motif in the 3'UTR of hGRbeta mRNA results not only in increased mRNA stability, but also in increased receptor protein expression, which may contribute to glucocorticoid resistance. A similar role is suggested for two other AUUUA motifs in the 3'UTR of hGRbeta mRNA and for the 10 AUUUA motifs that are present in the 3'UTR of hGRalpha. Published by Elsevier Science Inc. C1 NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Schaaf, MJM (reprint author), NIEHS, Lab Signal Transduct, NIH, 111 Alexander Dr,POB 12233, Res Triangle Pk, NC 27709 USA. NR 44 TC 80 Z9 83 U1 0 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0039-128X J9 STEROIDS JI Steroids PD JUN PY 2002 VL 67 IS 7 BP 627 EP 636 AR PII S0039-128X(02)00015-6 DI 10.1016/S0039-128X(02)00015-6 PG 10 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 564UE UT WOS:000176331300012 PM 11996936 ER PT J AU Wong, TY Klein, R Sharrett, AR Nieto, FJ Boland, LL Couper, DJ Mosley, TH Klein, BEK Hubbard, LD Szklo, M AF Wong, TY Klein, R Sharrett, AR Nieto, FJ Boland, LL Couper, DJ Mosley, TH Klein, BEK Hubbard, LD Szklo, M TI Retinal microvascular abnormalities and cognitive impairment in middle-aged persons - The atherosclerosis risk in communities study SO STROKE LA English DT Article DE cognitive disorders; dementia, vascular; hypertension; retina; retinal diseases ID JAPANESE PROVINCIAL CITY; POPULATION-BASED COHORT; CEREBRAL BLOOD-FLOW; ALZHEIMERS-DISEASE; BRAIN INFARCTION; DEMENTIA; STROKE; PRESSURE; DECLINE; MICROCIRCULATION AB Background and Purpose-Cerebral microvascular disease has been hypothesized to contribute to cognitive impairment, but few clinical data are available. Here, we examine the relation of retinal microvascular abnormalities with cognitive function in middle-aged persons free of stroke. Methods-The Atherosclerosis Risk in Communities Study is a population-based study with examinations every 3 years from 1987 through 1998. At visit 3, when participants were 51 to 70 years of age, retinal photographs were obtained and evaluated for retinal microvascular abnormalities according to standardized protocols. Cognitive function was assessed with standardized tests (Delayed Word Recall Test, Digit Symbol Subtest, and Word Fluency Test) at visits 2 and 4 and averaged for analysis. Persons with stroke or taking central nervous system-relevant medications were -excluded, leaving 8734 with data for this study. Results-After education, diabetes mellitus, blood pressure, carotid intima-media thickness, and other risk factors were controlled for, retinopathy was associated with lower cognitive test scores. The. adjusted odds ratios for persons with Delayed Word Recall scores 2 SD or lower than the mean were 2.60 [95% confidence interval (CI), 1.30 to 2.91] for any retinopathy, 3.00 (95% CI, 1.81 to 4.98) for microaneurysms, 3.39 (95% CI, 1.99 to 5.78) for retinal hemorrhage, and 3.07 (95% CI, 1.53 to 6.17) for soft exudates. Results were similar for the other 2 cognitive tests and in people with and without diabetes and hypertension. Conclusions-Retinopathy is independently associated with poorer cognitive function in middle-aged persons without stroke, suggesting that cerebral microvascular disease may contribute to the development of cognitive impairment. C1 Natl Univ Singapore, Dept Ophthalmol, Singapore 119260, Singapore. Univ Wisconsin, Dept Ophthalmol, Madison, WI USA. Johns Hopkins Univ, Sch Publ Hlth, Dept Epidemiol, Baltimore, MD USA. Natl Univ Singapore, Singapore Natl Eye Ctr, Singapore, Singapore. Natl Univ Singapore, Dept Ophthalmol, Singapore, Singapore. Natl Heart Lung & Blood Inst, NIH, Bethesda, MD USA. Univ Minnesota, Div Epidemiol, Minneapolis, MN USA. Univ N Carolina, Dept Biostat, Chapel Hill, NC USA. Univ Mississippi, Med Ctr, Dept Med, Jackson, MS USA. RP Wong, TY (reprint author), Natl Univ Singapore, Dept Ophthalmol, 10 Kent Ridge Crescent, Singapore 119260, Singapore. OI Klein, Ronald/0000-0002-4428-6237 FU NHLBI NIH HHS [N01-HC-55019, N01-HC-35125, N01-HC-35126, N01-HC-55015, N01-HC-55016, N01-HC-55018, N01-HC-55020, N01-HC-55021, N01-HC-55022] NR 40 TC 129 Z9 130 U1 0 U2 7 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0039-2499 J9 STROKE JI Stroke PD JUN PY 2002 VL 33 IS 6 BP 1487 EP 1492 DI 10.1161/01.STR.0000016789.56668.43 PG 6 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA 561VT UT WOS:000176164300021 PM 12052979 ER PT J AU Backman, C Morales, M AF Backman, C Morales, M TI Acute methamphetamine administration upregulates NGFI-B mRNA expression in the striatum: Co-localization with C-FOS immunoreactivity SO SYNAPSE LA English DT Article DE Nur77; distribution; transcription factors; brain; basal ganglia ID MESSENGER-RNA EXPRESSION; FACTOR-INDUCIBLE-B; RAT FOREBRAIN; TRANSCRIPTION FACTOR; DOPAMINE-RECEPTORS; BASAL GANGLIA; INDUCTION; COCAINE; AMPHETAMINE; PATTERNS AB In this study, the effects of acute methamphetamine administration on expression of the nuclear transcription factor NGFI-B mRNA and its co-localization with c-Fos immunoreactivity in the striatum were evaluated in animals receiving a single dose of methamphetamine (4 mg/kg) given at 2 or 6 h prior to perfusion. All animals received a daily saline injection for 6 days prior to methamphetamine treatment. We have found that, similar to c-fos activation, NGFI-B mRNA levels were significantly increased 2 h after methamphetamine treatment and returned to basal levels 6 h later. Induction of NGFI-B mRNA levels by methamphetamine was highest in central striatum as compared to the dorsomedial distribution pattern observed in control animals. After acute methamphetamine treatment, the distribution pattern of NGFI-B mRNA upregulation. was very similar to that of methamphetamine induced c-Fos immunoreactivity. However, co-localization studies with c-Fos immunoreactivity showed that not all NGFI-B-positive cells contained c-Fos after methamphetamine treatment. Forty-five percent of all NGFI-B mRNA expressing neurons contained c-Fos immunoreactivity in the dorsomedial striatum as compared to 60% in central and 35% in ventrolateral striatum. This study provides a detailed description of the differential spatial and temporal modulation of NGFI-B and c-Fos expression in the striatum by acute methamphetamine treatment over time. Published 2002 Wiley-Liss, Inc. C1 NIDA, NIH, Cellular Neurobiol Dept, Baltimore, MD 21224 USA. RP Backman, C (reprint author), NIDA, NIH, Cellular Neurobiol Dept, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI backman, cristina/C-1276-2013 NR 25 TC 13 Z9 14 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-4476 J9 SYNAPSE JI Synapse PD JUN 1 PY 2002 VL 44 IS 3 BP 158 EP 165 DI 10.1002/syn.10065 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 540VZ UT WOS:000174952400006 PM 11954047 ER PT J AU Jordan, IK Rogozin, IB Wolf, YI Koonin, EV AF Jordan, IK Rogozin, IB Wolf, YI Koonin, EV TI Microevolutionary genomics of bacteria SO THEORETICAL POPULATION BIOLOGY LA English DT Article ID PATHOGEN HELICOBACTER-PYLORI; CHLAMYDIA-PNEUMONIAE; ESCHERICHIA-COLI; SUBSTITUTION RATES; MAMMALIAN GENES; DUPLICATE GENES; COG DATABASE; EVOLUTION; SEQUENCE; PROTEINS AB The availability of multiple complete genome sequences from the same species can facilitate attempts to systematically address basic questions in genome evolution. We refer to such efforts as "microevolutionary genomics". We report the results of comparative analyses of complete intraspecific genome (and proteome) sequences from four bacterial species-Chlamydophila pneumoniae, Escherichia coli, Helicobacter pylori and Neisseria, meningitidis. Comparisons of average synonymous (K-s) and nonsynonymous (K.) substitution rates were used to assess the influence of various biological factors on the rate of protein evolution. For example, E. coli experiences the most intense purifying selection of the species analyzed, and this may be due to the relatively larger population size of this species. In addition, essential genes were shown to be more evolutionarily conserved than nonessential genes in E. cofi and duplicated genes have higher rates of evolution than unique genes for all species studied except C. pneumoniae. Different functional categories of genes were shown to evolve at significantly different rates emphasizing the role of category-specific functional constraints in determining evolutionary rates. Finally, functionally characterized genes tend to be conserved between strains, while uncharacterized genes are over-represented among the unique, strain-specific genes. This suggests the possibility that nonessential genes are responsible for driving the evolutionary diversification between strains. (C) 2002 Elsevier Science (USA). C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. RP Jordan, IK (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bldg 38A,8600 Rockville Pike, Bethesda, MD 20894 USA. NR 63 TC 65 Z9 68 U1 0 U2 4 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0040-5809 J9 THEOR POPUL BIOL JI Theor. Popul. Biol. PD JUN PY 2002 VL 61 IS 4 BP 435 EP 447 DI 10.1006/tpbi.2002.1588 PG 13 WC Ecology; Evolutionary Biology; Genetics & Heredity SC Environmental Sciences & Ecology; Evolutionary Biology; Genetics & Heredity GA 589CB UT WOS:000177739500006 PM 12167363 ER PT J AU Hashimoto, K Peebles, RS Sheller, JR Jarzecka, K Furlong, J Mitchell, DB Hartert, TV Graham, BS AF Hashimoto, K Peebles, RS Sheller, JR Jarzecka, K Furlong, J Mitchell, DB Hartert, TV Graham, BS TI Suppression of airway hyperresponsiveness induced by ovalbumin sensitisation and RSV infection with Y-27632, a Rho kinase inhibitor SO THORAX LA English DT Article ID RESPIRATORY SYNCYTIAL VIRUS; SMOOTH-MUSCLE CONTRACTION; CALCIUM SENSITIZATION; CA2+ SENSITIZATION; MYOSIN; ASTHMA; PHOSPHORYLATION; PHOSPHATASE; MEDIATORS; TRACHEAL AB Background: Smooth muscle contraction is one of the hallmarks of asthma. A recently developed pyridine derivative, Y-27632, a selective Rho kinase inhibitor, has been reported to inhibit the smooth muscle contraction of human and animal trachea in ex vivo systems but its effect in animal models of airway hyperresponsiveness (AHR) has not been examined. The purpose of this study was to evaluate the effect of Y-27632 in a murine model of allergic and virally induced AHR. Methods: Baseline lung resistance and methacholine induced AHR were measured in mice sensitised to ovalbumin (OVA) and also in mice infected with respiratory syncytial virus (RSV) following ovalbumin sensitisation (OVA/RSV). Results: Time course and dose ranging experiments indicated that 30 mg/kg Y-27632 given by gavage 2 hours before methacholine challenge significantly reduced baseline lung resistance and prevented AHR in OVA sensitised mice. Y-27632 also suppressed AHR induced by the bronchospastic agent serotonin in OVA sensitised mice and prevented methacholine induced AHR in OVA/RSV mice. Conclusions: These results suggest that the signalling pathway mediated through Rho kinase may have an important role in bronchial smooth muscle tone in allergen induced and virus induced AHR and should be considered as a novel target for asthma treatment. C1 NIAID, VRC, Viral Pathogenesis Lab & Clin Trials Core, NIH, Bethesda, MD 20892 USA. Vanderbilt Univ, Sch Med, Dept Med, Nashville, TN 37232 USA. Fukushima Med Univ, Dept Microbiol, Sch Med, Fukushima 9601295, Japan. RP Graham, BS (reprint author), NIAID, VRC, Viral Pathogenesis Lab & Clin Trials Core, NIH, Bldg 40 Rm 2502 40 Convent Dr MSC 3017, Bethesda, MD 20892 USA. FU NHLBI NIH HHS [K08-HL-03730]; NIAID NIH HHS [R01-AI-45512]; NIGMS NIH HHS [GM 15431] NR 29 TC 39 Z9 39 U1 0 U2 2 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0040-6376 J9 THORAX JI Thorax PD JUN PY 2002 VL 57 IS 6 BP 524 EP 527 DI 10.1136/thorax.57.6.524 PG 4 WC Respiratory System SC Respiratory System GA 559LW UT WOS:000176028500010 PM 12037228 ER PT J AU Chriqui, JF Frosh, MM Fues, LA el Arculli, R Stillman, FA AF Chriqui, JF Frosh, MM Fues, LA el Arculli, R Stillman, FA TI State laws on youth access to tobacco: an update, 1993-1999 SO TOBACCO CONTROL LA English DT Letter C1 MayaTech Corp, Silver Spring, MD 20910 USA. NCI, Bethesda, MD 20892 USA. Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Inst Global Tobacco Control, Baltimore, MD USA. RP Chriqui, JF (reprint author), MayaTech Corp, 8737 Colesville Rd,7th Floor, Silver Spring, MD 20910 USA. FU NCI NIH HHS [N2-PC-75007] NR 9 TC 4 Z9 4 U1 4 U2 4 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0964-4563 J9 TOB CONTROL JI Tob. Control PD JUN PY 2002 VL 11 IS 2 BP 163 EP 164 DI 10.1136/tc.11.2.163 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 562XV UT WOS:000176225000024 PM 12035016 ER PT J AU Hamadeh, HK Bushel, PR Jayadev, S Martin, K DiSorbo, O Sieber, S Bennett, L Tennant, R Stoll, R Barrett, JC Blanchard, K Paules, RS Afshari, CA AF Hamadeh, HK Bushel, PR Jayadev, S Martin, K DiSorbo, O Sieber, S Bennett, L Tennant, R Stoll, R Barrett, JC Blanchard, K Paules, RS Afshari, CA TI Gene expression analysis reveals chemical-specific profiles SO TOXICOLOGICAL SCIENCES LA English DT Article DE gene expression; toxicogenomics; DNA arrays; classification; rat liver; pattern recognition ID PEROXISOME PROLIFERATOR; CELL-PROLIFERATION; ENZYME-INDUCTION; CDNA MICROARRAYS; RAT-LIVER; TOXICOLOGY; TOXICITY; TOXICOGENOMICS; MECHANISMS; PROMOTION AB The application of gene expression profiling technology to examine multiple genes and signaling pathways simultaneously promises a significant advance in understanding toxic mechanisms to ultimately aid in protection of public health. Public and private efforts in the new field of toxicogenomics are focused on populating databases with gene expression profiles of compounds where toxicological and pathological endpoints are well characterized. The validity and utility of a toxicogenomics is dependent on whether gene expression profiles that correspond to different chemicals can be distinguished. The principal hypothesis underlying a toxicogenomic or pharmacogenomic strategy is that chemical-specific patterns of altered gene expression will be revealed using high-density microarray analysis of tissues from exposed organisms. Analyses of these patterns should allow classification of toxicants and provide important mechanistic insights. This report provides a verification of this hypothesis. Patterns of gene expression corresponding to liver tissue derived from chemically exposed rats revealed similarity in gene expression profiles between animals treated with different agents from a common class of compounds, peroxisome proliferators [clofibrate (ethyl-p-chlorophenoxyisobutyrate), Wyeth 14,643 ([4-chloro-6(2,3-xylidino)-2-pyrimidinylthio]acetic acid), and gemfibrozil (5-2[2,5-dimethyl-phenoxy] 2-2-dimethylpentanoic acid)], but a very distinct gene expression profile was produced using a compound from another class, enzyme inducers (phenobarbital). C1 NIEHS, Res Triangle Pk, NC 27709 USA. Boehringer Ingelheim Pharmaceut Inc, Ridgefield, CT 06877 USA. RP Afshari, CA (reprint author), NIEHS, POB 12233,MD2-04, Res Triangle Pk, NC 27709 USA. NR 32 TC 298 Z9 318 U1 0 U2 19 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 J9 TOXICOL SCI JI Toxicol. Sci. PD JUN PY 2002 VL 67 IS 2 BP 219 EP 231 DI 10.1093/toxsci/67.2.219 PG 13 WC Toxicology SC Toxicology GA 556MD UT WOS:000175852800009 PM 12011481 ER PT J AU Hamadeh, HK Bushel, PR Jayadev, S DiSorbo, O Bennett, L Li, LP Tennant, R Stoll, R Barrett, JC Paules, RS Blanchard, K Afshari, CA AF Hamadeh, HK Bushel, PR Jayadev, S DiSorbo, O Bennett, L Li, LP Tennant, R Stoll, R Barrett, JC Paules, RS Blanchard, K Afshari, CA TI Prediction of compound signature using high density gene expression profiling SO TOXICOLOGICAL SCIENCES LA English DT Article DE toxicogenomics; gene expression database; discriminant genes; prediction; algorithms; DNA arrays ID PEROXISOME PROLIFERATORS; ANTIEPILEPTIC DRUGS; RODENT CARCINOGENS; MICROARRAY; PATTERNS; SAR; CYTOCHROME-P-450; CLASSIFICATION; IDENTIFICATION; TOXICOGENOMICS AB DNA microarrays, used to measure the gene expression of thousands of genes simultaneously, hold promise for future application in efficient screening of therapeutic drugs. This will be aided by the development and population of a database with gene expression profiles corresponding to biological responses to exposures to known compounds whose toxicological and pathological endpoints are well characterized. Such databases could then be interrogated, using profiles corresponding to biological responses to drugs after developmental or environmental exposures. A positive correlation with an archived profile could lead to some knowledge regarding the potential effects of the tested compound or exposure. We have previously shown that cDNA microarrays can be used to generate chemical-specific gene expression profiles that can be distinguished across and within compound classes, using clustering, simple correlation, or principal component analyses. In this report, we test the hypothesis that knowledge can be gained regarding the nature of blinded samples, using an initial training set comprised of gene expression profiles derived from rat liver exposed to clofibrate, Wyeth 14,643, gemfibrozil, or phenobarbital for 24 h or 2 weeks of exposure. Highly discriminant genes were derived from our database training set using approaches including linear discriminant analysis (LDA) and genetic algorithm/K-nearest neighbors (GA/KNN). Using these genes in the analysis of coded liver RNA samples derived from 24-h, 3-day, or 2-week exposures to phenytoin, diethylhexylpthalate, or hexobarbital led to successful prediction of whether these samples were derived from livers of rats exposed to enzyme inducers or to peroxisome proliferators. This validates our initial hypothesis and lends credibility to the concept that the further development of a gene expression database for chemical effects will greatly enhance the hazard identification processes. C1 NIEHS, Res Triangle Pk, NC 27709 USA. Boehringer Ingelheim Pharmaceut Inc, Ridgefield, CT 06877 USA. RP Afshari, CA (reprint author), NIEHS, POB 12233,MD2-04, Res Triangle Pk, NC 27709 USA. NR 37 TC 190 Z9 205 U1 0 U2 8 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 J9 TOXICOL SCI JI Toxicol. Sci. PD JUN PY 2002 VL 67 IS 2 BP 232 EP 240 DI 10.1093/toxsci/67.2.232 PG 9 WC Toxicology SC Toxicology GA 556MD UT WOS:000175852800010 PM 12011482 ER PT J AU Greco, VA Byrne, KM Procter, JL Stroncek, DF AF Greco, VA Byrne, KM Procter, JL Stroncek, DF TI Detection of antibodies in acid eluates with the gel microcolumn assay SO TRANSFUSION LA English DT Article ID ANTIGLOBULIN-TEST; TUBE TECHNIQUE; AGGLUTINATION; COLUMN AB BACKGROUND: Gel microcolumns can be used to detect unexpected serum antibodies and to determine ABO blood group and Rh phenotype. DATs can also be performed with this system. The purpose of this study was to compare the gel microcolumn to the tub IAT using anti-IgG for the detection of antibodies eluted from RBCs. STUDY DESIGN AND METHODS: Acid eluates were prepared from 30 peripheral blood and 41 umbilical cord blood samples. Twelve of the 71 eluates made were from control samples (known DAT negative). Specificities of eluted antibodies were determined by both tube and gel assays with a three-cell screen plus A, and B cells, as determined by blood type. RESULTS: Ten of 30 peripheral blood eluates were reactive in both assays. Eighteen were nonreactive in both assays, and two from patients with autoimmune hemolytic anemia were reactive by gel assays and nonreactive by tube assays. Thirty three of the 41 cord blood eluates were reactive in both assays. Eluates from 2 of the 35 DAT-positive samples reacted with A, and B cells by the tube method but were nonreactive by the gel method. Of the 33 cord blood eluates that were reactive by both assays, antibody specificity differed for two samples. When tested by tube assay, these eluates reacted with both A, and B cells, whereas the same eluates tested by gel assay showed one reacting with only A, cells and the other with only B cells. CONCLUSIONS: Results of testing eluates in gel assays were similar to those obtained in tube assays. The gel assays may be better at detecting antibodies eluted from RBCs from patients with autoimmune hemolytic anemia, and tube assays may be better at detecting isohemagglutinins eluted from umbilical cord blood. C1 NIH, Warren G Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. RP Byrne, KM (reprint author), NIH, Warren G Magnuson Clin Ctr, Dept Transfus Med, Bldg 10,Room 1C711,10 Ctr Dr MSC 1184, Bethesda, MD 20892 USA. NR 14 TC 6 Z9 8 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 JUN PY 2002 VL 42 IS 6 BP 698 EP 701 DI 10.1046/j.1537-2995.2002.00108.x PG 4 WC Hematology SC Hematology GA 568TJ UT WOS:000176559400007 PM 12147021 ER PT J AU Chambers, DR Procter, J Muratova, O Byrne, K Keister, D Shanks, D Magill, A Stroncek, D AF Chambers, DR Procter, J Muratova, O Byrne, K Keister, D Shanks, D Magill, A Stroncek, D TI In vitro RBC exposure to Plasmodium falciparum has no effect on RBC antigen expression SO TRANSFUSION MEDICINE LA English DT Article DE erythrocyte antigens; malarial anaemia; Plasmodium falciparum ID MALARIA; CELL; CHILDREN; ANEMIA AB Severe malarial anaemia is a leading cause of death in African children younger than 3 years of age who are infected with Plasmodium falciparum . The pathogenesis of this anaemia is not understood. The purpose of this study was to determine if P. falciparum induces changes in RBC membranes that contribute to the immune destruction of RBCs. RBCs were collected from healthy subjects and tested using standard haemagglutination assays for 45 antigens representing 21 blood group systems/collections before and after exposure to P. falciparum , strain FVO. Lectins were used to determine whether crypt or neoantigens were expressed on the RBC membrane. Polybrene was used to detect changes in sialic acid. RBCs were cultured in vitro with and without the parasite, and blinded serologic studies were completed. CD35 (complement receptor 1), CD55 (decay-accelerating factor), CD59 (membrane inhibitor of reactive lysis) and CD47 (integrin-associated protein) flow cytometric assays were compared for infected and uninfected RBCs. The percentage of parasitaemia was determined using Giemsa-stained thin blood films. Two (Ch, Lu-b ) of the 45 antigens had differing strengths of agglutination between infected and uninfected RBCs, but these differences were resolved with a second source of antisera. Forty-three antigens showed no significant differences in the strength of agglutination between the infected and uninfected RBCs. Lectin and polybrene testing showed no differences. CD35, CD55, CD59 and CD47 levels showed no significant differences. P. falciparum does not appear to alter the expression of classified immunogenic antigens on the RBC membrane in this in vitro system. The pathogenesis of the haemolytic episode that occurs in these children remains unclear. C1 NIH, Warren G Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. NIAID, NIH, Bethesda, MD 20892 USA. USAMC, AFRMIS, Bangkok, Thailand. RP Stroncek, D (reprint author), NIH, Warren G Magnuson Clin Ctr, Dept Transfus Med, Bldg 10,Room 1C711,10 Ctr Dr,MSC-1184, Bethesda, MD 20892 USA. NR 16 TC 3 Z9 3 U1 0 U2 1 PU BLACKWELL PUBLISHING LTD PI OXFORD PA 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND SN 0958-7578 J9 TRANSFUSION MED JI Transfus. Med. PD JUN PY 2002 VL 12 IS 3 BP 213 EP 219 DI 10.1046/j.1365-3148.2002.00375.x PG 7 WC Hematology SC Hematology GA 564CH UT WOS:000176297100007 PM 12071878 ER PT J AU Yang, D Biragyn, A Kwak, LW Oppenheim, JJ AF Yang, D Biragyn, A Kwak, LW Oppenheim, JJ TI Mammalian defensins in immunity: more than just microbicidal SO TRENDS IN IMMUNOLOGY LA English DT Article ID IMMATURE DENDRITIC CELLS; HUMAN NEUTROPHIL DEFENSINS; ANTIMICROBIAL PEPTIDES; HOST-DEFENSE; MAST-CELLS; HUMAN BETA-DEFENSIN-3; ANTITUMOR IMMUNITY; ADAPTIVE IMMUNITY; ALPHA-DEFENSINS; BETA-DEFENSINS AB Mammalian defensins are small, cationic, antimicrobial peptides encoded by the host that are considered to be important antibiotic-like effectors of innate immunity. By using chemokine receptors on dendritic cells and T cells, defensins might also contribute to the regulation of host adaptive immunity against microbial invasion. Defensins have considerable immunological adjuvant activity and linkage of beta-defensins or selected chemokines to an idiotypic lymphoma antigen has yielded potent antitumor vaccines. The functional overlap between defensins and chemokines is reinforced by reports that some chemokines have antimicrobial activities. Although showing similarity in activity and overall tertiary structure, the evolutionary relationship between defensins and chemokines remains to be determined. C1 NCI, Canc Res Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NCI, Expt Transplantat & Immunol Branch, Bethesda, MD 20892 USA. RP Oppenheim, JJ (reprint author), NCI, Canc Res Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NR 47 TC 449 Z9 503 U1 7 U2 30 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4906 J9 TRENDS IMMUNOL JI Trends Immunol. PD JUN PY 2002 VL 23 IS 6 BP 291 EP 296 AR PII S1471-4906(02)02246-9 DI 10.1016/S1471-4906(02)02246-9 PG 6 WC Immunology SC Immunology GA 559GY UT WOS:000176019500007 PM 12072367 ER PT J AU Chanock, S Wacholder, S AF Chanock, S Wacholder, S TI One gene and one outcome? No way SO TRENDS IN MOLECULAR MEDICINE LA English DT Article ID SINGLE-NUCLEOTIDE POLYMORPHISMS; COMPLEX DISEASES; HUMAN-GENOME; RISK; SUSCEPTIBILITY; EPIDEMIOLOGY; CANCER; FUTURE; BRCA1 AB The world of molecular diagnostics is undergoing major change because of both technical advances and the availability of rapidly expanding genetic databases generated by the study of human genomics. These resources comprise an extraordinary opportunity to decipher the biological importance of genetic aberrations, and link our understanding with clinical utility. The challenge lies in sorting through the information and developing effective strategies to advance molecular diagnostics. C1 NCI, Sect Genom Variat, Pediat Oncol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Chanock, S (reprint author), NCI, Sect Genom Variat, Pediat Oncol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. NR 23 TC 24 Z9 25 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4914 J9 TRENDS MOL MED JI Trends Mol. Med PD JUN PY 2002 VL 8 IS 6 BP 266 EP 269 AR PII S1471-4914(02)02356-0 DI 10.1016/S1471-4914(02)02356-0 PG 4 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 557VB UT WOS:000175928000007 PM 12067611 ER PT J AU Negm, RS Verma, M Srivastava, S AF Negm, RS Verma, M Srivastava, S TI The promise of biomarkers in cancer screening and detection SO TRENDS IN MOLECULAR MEDICINE LA English DT Review ID SINGLE NUCLEOTIDE POLYMORPHISMS; GENE-EXPRESSION; LUNG-CANCER; ESOPHAGEAL ADENOCARCINOMA; ABERRANT METHYLATION; COLORECTAL-CANCER; BLADDER CANCERS; TYROSINE KINASE; ACUTE-LEUKEMIA; EARLY EVENT AB Despite the recent decline in the incidence of cancer, long-term mortality rates remain unchanged. One of the most important factors in the survival of cancer is detection at an early stage. Clinical assays that detect the early events of cancer offer an opportunity to intervene and prevent cancer progression. Biomarkers are important molecular signatures of the phenotype of a cell that aid in early cancer detection and risk assessment. Although new information and technologies are clearly important for new biomarker discovery, we face major hurdles in translating new findings into clinical application. Here, we discuss examples of recent advances and limitations in cancer biomarker identification and validation, and the implications for cancer prevention. C1 NCI, Canc Biomarkers Res Grp, Div Canc Prevent, NIH, Rockville, MD 20852 USA. RP Srivastava, S (reprint author), NCI, Canc Biomarkers Res Grp, Div Canc Prevent, NIH, Rockville, MD 20852 USA. NR 55 TC 62 Z9 69 U1 0 U2 7 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4914 J9 TRENDS MOL MED JI Trends Mol. Med PD JUN PY 2002 VL 8 IS 6 BP 288 EP 293 AR PII S1471-4914(02)02353-5 DI 10.1016/S1471-4914(02)02353-5 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 557VB UT WOS:000175928000011 PM 12067615 ER PT J AU Nutman, TB AF Nutman, TB TI Future directions for vaccine-related onchocerciasis research SO TRENDS IN PARASITOLOGY LA English DT Editorial Material ID FILARIAL NEMATODES; VOLVULUS; IMMUNITY; DISEASE AB Although the general perception has been that Onchocerca volvulus infection is well on its way towards extinction, it is becoming increasingly clear that without an additional therapeutic or preventative modality (separate from ivermectin distribution or vector control), elimination of onchocercal infection will remain an unfulfilled goal. Vaccination aimed at preventing infection (directed toward infective-stage or adult worms), transmission and/or pathology (directed toward skin microfilariae) could provide the necessary' final punch' to knockout human infection with O. volvulus altogether. Thus, efforts toward the development of a useful vaccine need to be redoubled rather than shelved, and, to this end, major gaps in our knowledge need to be filled. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Nutman, TB (reprint author), NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. NR 21 TC 15 Z9 15 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1471-4922 J9 TRENDS PARASITOL JI Trends Parasitol. PD JUN PY 2002 VL 18 IS 6 BP 237 EP 239 AR PII S1471-4922(02)02310-3 DI 10.1016/S1471-4922(02)02310-3 PG 3 WC Parasitology SC Parasitology GA 555FW UT WOS:000175784600001 PM 12036727 ER PT J AU Sapone, A Paolini, M Biagi, GL Cantelli-Forti, G Gonzalez, FJ AF Sapone, A Paolini, M Biagi, GL Cantelli-Forti, G Gonzalez, FJ TI The pressing need for combined genotype-phenotype analysis in clinical practice SO TRENDS IN PHARMACOLOGICAL SCIENCES LA English DT Letter ID DRUG DEVELOPMENT; GENOME C1 Univ Bologna, Dept Pharmacol, Pharmacogenom Unit, I-40126 Bologna, Italy. Univ Texas, Dept Prevent Med & Commun Hlth, Med Branch, Galveston, TX 77555 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. RP Sapone, A (reprint author), Univ Bologna, Dept Pharmacol, Pharmacogenom Unit, Via Irnerio 48, I-40126 Bologna, Italy. RI Sapone, Andrea/E-6704-2013; OI Sapone, Andrea/0000-0001-8496-6977 NR 11 TC 4 Z9 4 U1 0 U2 0 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0165-6147 J9 TRENDS PHARMACOL SCI JI Trends Pharmacol. Sci. PD JUN PY 2002 VL 23 IS 6 BP 260 EP 261 AR PII S0165-6147(02)02047-3 DI 10.1016/S0165-6147(02)02047-3 PG 2 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 559GU UT WOS:000176019000006 PM 12084629 ER PT J AU Propert, KJ Alexander, RB Nickel, JC Kusek, JW Litwin, MS Landis, JR Nyberg, LM Schaeffer, AJ AF Propert, KJ Alexander, RB Nickel, JC Kusek, JW Litwin, MS Landis, JR Nyberg, LM Schaeffer, AJ CA Chronic Prostatitis Collaborative TI Design of a multicenter randomized clinical trial for chronic prostatitis/chronic pelvic pain syndrome SO UROLOGY LA English DT Article ID CHRONIC BACTERIAL PROSTATITIS; ANTIBIOTIC-THERAPY; CIPROFLOXACIN; INFLAMMATION; ALPHA; MEN AB Objectives. To describe the study design and rationale for the first placebo-controlled, randomized clinical trial to be conducted by the Chronic Prostatitis Collaborative Research Network sponsored by the National Institutes of Health. Methods. The clinical trial will use a 2 by 2 factorial design to evaluate four treatments: placebo, tamsulosin hydrochloride alone, ciprofloxacin alone, and tamsulosin hydrochloride and ciprofloxacin combined. The primary objectives are to compare the efficacy and safety endpoints in patients with chronic prostatitis/chronic pelvic pain syndrome. The study population consists of men with symptoms of discomfort or pain in the pelvic region for at least a 3-month period. One hundred eighty-four participants will be treated for 6 weeks and followed up for an additional 6 weeks. The primary endpoint is the change in the overall National Institutes of Health Chronic Prostatitis Symptom Index from baseline to 6 weeks. Secondary endpoints include a patient-reported global assessment of response and white blood cell counts and cultures in urine, expressed prostatic secretions, and semen. The evaluation of changes from 6 to 12 weeks will be used to assess the durability of the response, Results. This trial was opened to enrollment in July 2001. The study is expected to require 14 months of accrual and follow-up. Conclusions. The issues considered in the design of this clinical trial may provide guidelines for future clinical trials in chronic prostatitis/chronic pelvic pain syndrome. (C) 2002, Elsevier Science Inc. C1 Univ Penn, Sch Med, Dept Biostat & Epidemiol, Philadelphia, PA 19104 USA. Univ Maryland, Baltimore, MD 21201 USA. Queens Univ, Kingston, ON, Canada. NIDDKD, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Sch Med, Los Angeles, CA USA. Northwestern Univ, Chicago, IL 60611 USA. RP Propert, KJ (reprint author), Univ Penn, Sch Med, Dept Biostat & Epidemiol, Blockley Hall,Room 614,423 Guardian Dr, Philadelphia, PA 19104 USA. RI Landis, J. Richard/A-9330-2010 FU NIDDK NIH HHS [R01 DK53730, R01 DK53732, R01 DK53734, R01 DK53736, R01 DK53746, R01 DK53752] NR 30 TC 49 Z9 57 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0090-4295 J9 UROLOGY JI UROLOGY PD JUN PY 2002 VL 59 IS 6 BP 870 EP 876 AR PII S0090-4295(02)01601-1 DI 10.1016/S0090-4295(02)01601-1 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA 554UG UT WOS:000175755100017 PM 12031372 ER PT J AU Platz, EA Smit, E Curhan, GC Nyberg, LM Giovannucci, E AF Platz, EA Smit, E Curhan, GC Nyberg, LM Giovannucci, E TI Prevalence of and racial/ethnic variation in lower urinary tract symptoms and noncancer prostate surgery in US men SO UROLOGY LA English DT Article ID HIGH-RISK GROUP; HYPERPLASIA; HYPERTROPHY; ETHNICITY; LIFE AB Objectives. To estimate the proportion of U.S. men affected by specific lower urinary tract symptoms (LUTS) and to assess whether the prevalence of LUTS varies by race/ethnicity. Methods. Included were 30+-year-old men who took part in the Third National Health and Nutrition Examination Survey. Men were asked whether they experienced nocturia, incomplete emptying, or hesitancy. Men 60+ years old were also asked whether they had a decreased urinary stream or had ever undergone noncancer prostate surgery. To obtain the estimated prevalences for the U.S. population, we applied sampling fraction weights, We calculated age-adjusted odds ratios (ORs) of 3+ symptoms or surgery by race/ethnicity using logistic regression analysis. Results. Only nocturia was common in 30 to 59-year-old men. Among men who had not had prostate surgery, 59.9% of men 60 to 69 years old and 75.1% of men 70+ years old had at least one symptom. All four symptoms were reported by 3.0% of men 60 to 69 years and 5.6% of men 70+ years old. Of the men 60 to 69 years old and men 70+ years old, 8.0% and 22.4%, respectively, reported having undergone surgery. In men 60+ years old, the age-adjusted OR for either having 3+ symptoms or surgery was 0.8 for non-Hispanic black men compared with non-Hispanic white men. The odds of having 3+ symptoms (OR = 1.6), but not surgery (OR = 1.1), appeared greater for Mexican-American men than for non-Hispanic white men. Conclusions. Specific LUTS are common in older U.S. men, Older black men were not more likely to have LUTS than were older white men. The apparent modestly higher prevalence of LUTS in older Mexican-American men requires additional study, (C) 2002, Elsevier Science Inc. C1 Johns Hopkins Bloomberg Sch Publ Hlth, Dept Epidemiol, Baltimore, MD 21205 USA. SUNY Buffalo, Sch Med, Dept Social & Prevent Med, Buffalo, NY 14260 USA. Massachusetts Gen Hosp, Dept Med, Boston, MA 02114 USA. NIDDKD, Div Kidney Urol & Hematol Dis, Bethesda, MD 20892 USA. Harvard Univ, Sch Publ Hlth, Dept Nutr, Boston, MA 02115 USA. Harvard Univ, Sch Publ Hlth, Dept Epidemiol, Boston, MA 02115 USA. Harvard Univ, Sch Med, Channing Lab, Dept Med, Boston, MA USA. Brigham & Womens Hosp, Boston, MA USA. RP Platz, EA (reprint author), Johns Hopkins Bloomberg Sch Publ Hlth, Dept Epidemiol, 615 N Wolfe St,Room E6143, Baltimore, MD 21205 USA. NR 25 TC 86 Z9 88 U1 1 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0090-4295 J9 UROLOGY JI UROLOGY PD JUN PY 2002 VL 59 IS 6 BP 877 EP 883 DI 10.1016/S0090-4295(01)01673-9 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA 554UG UT WOS:000175755100018 PM 12031373 ER PT J AU Fang, J Metter, EJ Landis, P Carter, HB AF Fang, J Metter, EJ Landis, P Carter, HB TI PSA velocity for assessing prostate cancer risk in men with PSA levels between 2.0 and 4.0 ng/ml SO UROLOGY LA English DT Article ID ANTIGEN LEVELS AB Objectives. To evaluate the use of prostate-specific antigen (PSA) velocity (PSAV) in assessing prostate cancer risk among men with PSA levels less than 4,0 ng/mL. Methods. The relative risk of, and cumulative probability of freedom from, prostate cancer by PSAV was evaluated in 89 male participants (21 with cancer and 68 controls) of a longitudinal aging study-the Baltimore Longitudinal Study of Aging (National Institute on Aging)-who had serial PSA levels between 2.0 and 4.0 ng/mL for at least 18 months. The relative risk was estimated from a Cox proportional hazards regression model, and the disease-free probability was determined by Kaplan-Meier survival analysis. The sensitivity and specificity of PSAV were calculated as a measure of test validity. Results. The sensitivity and specificity of a PSAV of 0.1 ng/mL per year was 81% and 50%, respectively, The relative risk of prostate cancer was 6.53 (range 1.90 to 22.51) when the PSAV was 0.1 ng/mL per year or more compared with a PSAV of less than 0.1 ng/mL per year (P = 0.0029). At 10 years, the cumulative probability of freedom from prostate cancer was 97.1% (range 91.4% to 100%) and 35.2% (range 14.0% to 56.4%) when the PSAV was less than and greater than 0.1 ng/mL per year, respectively. Survival curves began to differ at less than 5 years after the baseline PSAV determination. Conclusions. The association between PSAV and the subsequent risk of prostate cancer suggests that the PSAV may be useful in the risk assessment of men with lower PSA levels. (C) 2002, Elsevier Science Inc. C1 Johns Hopkins Univ, Johns Hopkins Hosp, Dept Urol, Sch Med,James Buchanan Brady Urol Inst, Baltimore, MD 21287 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Carter, HB (reprint author), Johns Hopkins Univ, Johns Hopkins Hosp, Dept Urol, Sch Med,James Buchanan Brady Urol Inst, 403 Marburg,600 N Wolfe St, Baltimore, MD 21287 USA. FU NCI NIH HHS [CA58236] NR 15 TC 50 Z9 52 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0090-4295 J9 UROLOGY JI UROLOGY PD JUN PY 2002 VL 59 IS 6 BP 889 EP 893 AR PII S0090-4295(02)01646-1 DI 10.1016/S0090-4295(02)01646-1 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 554UG UT WOS:000175755100020 PM 12031375 ER PT J AU Botos, I Mori, T Cartner, LK Boyd, MR Wlodawer, A AF Botos, I Mori, T Cartner, LK Boyd, MR Wlodawer, A TI Domain-swapped structure of a mutant of cyanovirin-N SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID VIRUS-INACTIVATING PROTEIN; STRUCTURE REFINEMENT; NATURAL-PRODUCTS; GP120; CYANOBACTERIUM; MECHANISM; BINDING; MODEL; BOND; CD4 AB Cyanovirin-N (CV-N) is a potent 11 kDa HIV-inactivating protein that binds with high affinity to the HIV surface envelope protein gp120. A double mutant P51S/S52P of CV-N was engineered by swapping two critical hinge-region residues Pro51 and Ser52. This mutant has biochemical and biophysical characteristics equivalent to the wild-type CV-N and its structure resembles that of wild-type CV-N. However, the mutant shows a different orientation in the hinge region that connects two domains of the protein. The observation that this double mutant crystallizes under a wide variety of conditions challenges some of the current hypotheses on domain swapping and on the role of hinge-region proline residues in domain orientation. The current structure contributes to the understanding of domain swapping in cyanovirins, permitting rational design of domain-swapped CV-N mutants. (C) 2002 Published by Elsevier Science (USA). C1 NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. NCI, Ctr Canc Res, Mol Targets Drug Discovery Program, Frederick, MD 21702 USA. NCI, Sci Applicat Int Corp, Frederick, MD 21702 USA. RP Wlodawer, A (reprint author), NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. NR 34 TC 22 Z9 22 U1 1 U2 4 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 31 PY 2002 VL 294 IS 1 BP 184 EP 190 AR PII S0006-291X(02)00455-2 DI 10.1016/S0006-291X(02)00455-2 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 563BU UT WOS:000176234500031 PM 12054761 ER PT J AU Parent, CA AF Parent, CA TI PI 3-kinases and PTEN: How opposites chemoattract SO CELL LA English DT Review ID TUMOR-SUPPRESSOR PTEN; NEUTROPHIL CHEMOTAXIS; CELLS; LOCALIZATION; ACTIVATION; MIGRATION AB Phosphatidylinositol lipids, such as PI(4,5)P-2 and PI(3,4,5)P-3, are key mediators in diverse intracellular signaling pathways. Two recent reports examine how the metabolism of these lipids by phosphatidylinositol 3-kinases and the PTEN 3-phosphoinositide phosphatase may coordinate G protein coupled signaling pathways during eukaryotic chemotaxis. C1 NCI, Cellular & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Natl Inst Gen Med Sci, PRAT Res Fellowship Program, NIH, Bethesda, MD 20892 USA. RP Parent, CA (reprint author), NCI, Cellular & Mol Biol Lab, NIH, 37 Convent Dr,MSC 4255,Bldg 37,Room 1E24, Bethesda, MD 20892 USA. NR 20 TC 0 Z9 0 U1 0 U2 0 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE,, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD MAY 31 PY 2002 VL 109 IS 5 BP 541 EP 544 PG 4 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 558GL UT WOS:000175957900002 ER PT J AU Yang, YL Dong, B Mittelstadt, PR Xiao, H Ashwell, JD AF Yang, YL Dong, B Mittelstadt, PR Xiao, H Ashwell, JD TI HIV tat binds Egr proteins and enhances Egr-dependent transactivation of the Fas ligand promoter SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; CD4(+) T-CELLS; NF-KAPPA-B; UP-REGULATION; TRANSCRIPTION FACTOR; CYCLOSPORINE-A; ANTIRETROVIRAL THERAPY; LYMPHOCYTE-ACTIVATION; MEDIATED APOPTOSIS; DOWN-REGULATION AB HIV Tat can enhance activation-induced up-regulation of Fas ligand (Fast,), which may contribute to T cell apoptosis in human immune deficiency virus (HIV)-infected individuals. We have assessed functional and physical interactions between Tat and the Egr family of transcription factors (Egr-1, -2, and -3), the latter two of which are major participants in activation-induced Fast, up-regulation. Here we report that whereas Tat itself has no effect on the Fast, promoter, it binds to Egr-2 and -3 and synergizes with them to superinduce expression of a Fast, promoter-driven reporter. A Tat molecule containing a single amino acid substitution that results in the loss of transactivation activity for the HIV long terminal repeat still binds Egr-3 but can no longer enhance Egr-mediated transactivation of the Fast, promoter. Furthermore, the mutated Tat acts as a dominant negative inhibitor, blocking the superinduction of Fast, caused by wild type Tat. Because Tat is present in virus-infected cells and in the serum of HIV-infected individuals, these results suggest that increased expression of Fast, in these circumstances may result from the cooperative activities of activation-induced Egrs and Tat. C1 NCI, Lab Immune Cell Biol, NIH, Bethesda, MD 20892 USA. Rockefeller Univ, Biochem & Mol Biol Lab, New York, NY 10021 USA. RP Ashwell, JD (reprint author), NCI, Lab Immune Cell Biol, NIH, Bldg 10,Rm 1B-40, Bethesda, MD 20892 USA. NR 68 TC 32 Z9 33 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 MAY 31 PY 2002 VL 277 IS 22 BP 19482 EP 19487 DI 10.1074/jbc.M201687200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800034 PM 11909874 ER PT J AU Wu, JF Li, JF Huang, KP Huang, FL AF Wu, JF Li, JF Huang, KP Huang, FL TI Attenuation of protein kinase C and cAMP-dependent protein kinase signal transduction in the neurogranin knockout mouse SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID LONG-TERM POTENTIATION; NITRIC-OXIDE MODIFICATION; RAT-BRAIN NEUROGRANIN; MAP KINASE; ADENYLYL-CYCLASE; SYNAPTIC PLASTICITY; SUBSTRATE RC3; LATE-PHASE; AREA CA1; PHOSPHORYLATION AB Neurogranin (Ng) is a brain-specific, postsynaptically located protein kinase C (PKC) substrate, highly expressed in the cortex, hippocampus, striatum, and amygdala. This protein is a Ca2+-sensitive calmodulin (CaM)-binding protein whose CaM-binding affinity is modulated by phosphorylation and oxidation. To investigate the role of Ng in neural function, a strain of Ng knockout mouse (KO) was generated. Previously we reported (Pak, J. H., Huang, F. L., Li, J., Balschun, D., Reymann, K. G., Chiang, C., Westphal, H., and Huang, K.-P. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 1123211237) that these KO mice displayed no obvious neuro-anatomical abnormality, but exhibited deficits in learning and memory and activation of Ca2+/CaM-dependent protein kinase II. In this report, we analyzed several downstream phosphorylation targets in phorbol 12-myristate 13-acetate- and forskolin-treated hippocampal slices from wild type (WT) and KO mice. Phorbol 12-myristate 13-acetate caused phosphorylation of Ng in WT mice and promoted the translocation of PKC from the cytosolic to the particulate fractions of both the WT and KO mice, albeit to a lesser extent in the latter. Phosphorylation of downstream targets, including mitogen-activated protein kinases, 90-kDa ribosomal S6 kinase, and the cAMP response element binding protein (CREB) was significantly attenuated in KO mice. Stimulation of hippocampal slices with forskolin also caused greater stimulation of protein kinase A (PKA) in the WT as compared with those of the KO mice. Again, phosphorylation of the downstream targets of PKA was attenuated in the KO mice. These results suggest that Ng plays a pivotal role in regulating both PKC- and PKA-mediated signaling pathways, and that the deficits in learning and memory of spatial tasks detected in the KO mice may be the result of defects in the signaling pathways leading to the phosphorylation of CREB. C1 NICHHD, Metab Regulat Sect, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Huang, FL (reprint author), NICHHD, Metab Regulat Sect, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. OI Li, Junfa/0000-0002-1930-9724 NR 48 TC 44 Z9 63 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 MAY 31 PY 2002 VL 277 IS 22 BP 19498 EP 19505 DI 10.1074/jbc.M109082200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800036 PM 11912190 ER PT J AU Kokoska, RJ Bebenek, K Boudsocq, F Woodgate, R Kunkel, TA AF Kokoska, RJ Bebenek, K Boudsocq, F Woodgate, R Kunkel, TA TI Low fidelity DNA synthesis by a Y family DNA polymerase due to misalignment in the active site SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN DINB1 GENE; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; POL-ETA; SULFOLOBUS-SOLFATARICUS; XERODERMA-PIGMENTOSUM; REVERSE-TRANSCRIPTASE; INDUCED MUTAGENESIS; ADAPTIVE MUTATION; ERROR-PRONE AB Sulfolobus solfataricus DNA polymerase IV (Dpo4) is a member of the Y family of DNA polymerases whose crystal structure has recently been solved. As a model for other evolutionarily conserved Y family members that perform translesion DNA synthesis and have low fidelity, we describe here the base substitution and frameshift fidelity of DNA synthesis by Dpo4. Dpo4 generates all 12 base-base mismatches at high rates, 11 of which are similar to those of its human homolog, DNA polymerase kappa This result is consistent with the Dpo4 structure, implying lower geometric selection for correct base pairs. Surprisingly, Dpo4 generates C-dCMP mismatches at an unusually high average rate and preferentially at cytosine Ranked by 5'-template guanine. Dpo4 also has very low frameshift fidelity and frequently generates deletions of even noniterated nucleotides, especially cytosine flanked by a W-template guanine. Both unusual features of error specificity suggest that Dpo4 can incorporate dNTP precursors when two template nucleotides are present in the active site binding pocket. These results have implications for mutagenesis resulting from DNA synthesis by Y family polymerases. C1 NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bethesda, MD 20892 USA. RP Kunkel, TA (reprint author), NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. NR 43 TC 83 Z9 84 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 31 PY 2002 VL 277 IS 22 BP 19633 EP 19638 DI 10.1074/jbc.M202021200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800053 PM 11919199 ER PT J AU Jaffe, EK Kervinen, J Martins, J Stauffer, F Neier, R Wlodawer, A Zdanov, A AF Jaffe, EK Kervinen, J Martins, J Stauffer, F Neier, R Wlodawer, A Zdanov, A TI Species-specific inhibition of porphobilinogen synthase by 4-oxosebacic acid SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID X-RAY STRUCTURE; ESCHERICHIA-COLI; PSEUDOMONAS-AERUGINOSA; ARTIFICIAL GENE; DEHYDRATASE; RESOLUTION; MECHANISM; MG(II); IONS AB Porphobilinogen synthase (PBGS) catalyzes the condensation of two molecules of 5-aminolevulinic acid (ALA), an essential step in tetrapyrrole biosynthesis. 4-Oxosebacic acid (4-OSA) and 4,7-dioxosebacic acid (4,7-DOSA) are bisubstrate reaction intermediate analogs for PBGS. We show that 4-OSA is an active site-directed irreversible inhibitor for Escherichia coli PBGS, whereas human, pea, Pseudomonas aeruginosa, and Bradyrhizobium japonicum PBGS are insensitive to inhibition by 4-OSA. Some variants of human PBGS (engineered to resemble E. coli PBGS) have increased sensitivity to inactivation by 4-OSA, suggesting a structural basis for the specificity. The specificity of 4-OSA as a PBGS inhibitor is significantly narrower than that of 4,7-DOSA. Comparison of the crystal structures for E. coli PBGS inactivated by 4-OSA versus 4,7-DOSA shows significant variation in the half of the inhibitor that mimics the second substrate molecule (A-side ALA). Compensatory changes occur in the structure of the active site lid, which suggests that similar changes normally occur to accommodate numerous hybridization changes that must occur at C3 of A-side ALA during the PBGS-catalyzed reaction. A comparison of these with other PBGS structures identifies highly conserved active site water molecules, which are isolated from bulk solvent and implicated as proton acceptors in the PBGS-catalyzed reaction. C1 Fox Chase Canc Ctr, Inst Canc Res, Philadelphia, PA 19111 USA. Univ Neuchatel, Dept Chem, CH-2007 Neuchatel, Switzerland. NCI, Macromol Crystallog Lab, NIH, Frederick, MD 21702 USA. RP Jaffe, EK (reprint author), Fox Chase Canc Ctr, Inst Canc Res, 7701 Burholme Ave, Philadelphia, PA 19111 USA. FU NCI NIH HHS [CA06927]; NIEHS NIH HHS [ES03654] NR 30 TC 22 Z9 24 U1 2 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 31 PY 2002 VL 277 IS 22 BP 19792 EP 19799 DI 10.1074/jbc.M201486200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800072 PM 11909869 ER PT J AU Soeth, E Thurber, DB Smith, CL AF Soeth, E Thurber, DB Smith, CL TI The viral transactivator E1A regulates the mouse mammary tumor virus promoter in an isoform- and chromatin-specific manner SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID LONG TERMINAL REPEAT; ADENOVIRUS E1A; GLUCOCORTICOID RECEPTOR; TRANSCRIPTION FACTOR; SACCHAROMYCES-CEREVISIAE; CELLULAR PROTEINS; BINDING PROTEIN; MMTV PROMOTER; ZINC-FINGER; ACTIVATION AB Proteins encoded by the adenovirus E1A gene regulate both cellular and viral genes to mediate effects on cell cycle, differentiation, and cell growth control. We have identified the mouse mammary tumor virus (MMTV) promoter as a target of E1A action and investigated the role nucleoprotein structure plays in its response to E1A. Both 12 and 13 S forms target the MMTV promoter when it has a disorganized and accessible chromatin configuration. However, whereas the 13 S form is stimulatory, the 12 S form is repressive. When the MMTV promoter adopts an organized and repressed chromatin structure, it is targeted only by the 13 S form, which stimulates it. Although evidence indicates that E1A interacts with the SWI/SNF remodeling complex, E1A had no effect on chromatin remodeling at the MMTV promoter in organized chromatin. Analysis of E1A mutants showed that stimulation of the MMTV promoter is mediated solely through conserved region 3 and does not require interaction with Rb, p300/CBP-associated factor, or CBP/p300. Imaging analysis showed that E1A colocalizes with MMTV sequences in vivo, suggesting that it functions directly at the promoter. These results indicate that E1A stimulates the MMTV promoter in a fashion independent of chromatin conformation and through a direct mechanism involving interaction with the basal transcription machinery. C1 NCI, Signal Transduct Grp, Lab Receptor Biol & Gene Express, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. RP Smith, CL (reprint author), NCI, Signal Transduct Grp, Lab Receptor Biol & Gene Express, Ctr Canc Res,NIH, Bldg 41,Rm B608,41 Lib Dr MSC 5055, Bethesda, MD 20892 USA. NR 52 TC 3 Z9 3 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 MAY 31 PY 2002 VL 277 IS 22 BP 19847 EP 19854 DI 10.1074/jbc.M200629200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800080 PM 11909860 ER PT J AU Gu, JG Fujibayashi, A Yamada, KM Sekiguchi, K AF Gu, JG Fujibayashi, A Yamada, KM Sekiguchi, K TI Laminin-10/11 and fibronectin differentially prevent apoptosis induced by serum removal via phosphatidylinositol 3-kinase/Akt- and MEK1/ERK-dependent pathways SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FOCAL ADHESION KINASE; ACTIVATED PROTEIN-KINASE; CELL-CYCLE PROGRESSION; EPITHELIAL-CELLS; INTEGRIN ALPHA-3-BETA-1; SIGNAL-TRANSDUCTION; MAMMARY EPITHELIUM; DEPENDENT PATHWAY; SURVIVAL PATHWAY; TYROSINE KINASE AB Cell adhesion to the extracellular matrix inhibits apoptosis, but the molecular mechanisms underlying the signals transduced by different matrix components are not well understood. Here, we examined integrin-mediated antiapoptotic signals from laminin-10/11 in comparison with those from fibronectin, the best characterized extracellular adhesive ligand. We found that the activation of protein kinase B/Akt in cells adhering to laminin-10/11 can rescue cell apoptosis induced by serum removal. Consistent with this, wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase, or ectopic expression of a dominant-negative mutant of Akt selectively accelerated cell death upon serum removal. In contrast to laminin-10/11, fibronectin rescued cells from serum depletion-induced apoptosis mainly through the extracellular signal-regulated kinase pathway. Cell survival on fibronectin but not laminin was significantly reduced by treatment with PD98059, a specific inhibitor of mitogen- or extracellular signal-regulated kinase kinase-1 (MEK1) and by expression of a dominant-negative mutant of MEK1. Laminin-10/11 was more potent than fibronectin in preventing apoptosis induced by serum depletion. These results, taken together, demonstrate laminin-10/11 potency as a survival factor and demonstrate that different extracellular matrix components can transduce distinct survival signals through preferential activation of subsets of multiple integrin-mediated signaling pathways. C1 Osaka Univ, Inst Prot Res, Div Prot Chem, Suita, Osaka 5650871, Japan. NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. RP Sekiguchi, K (reprint author), Osaka Univ, Inst Prot Res, Div Prot Chem, 3-2 Yamadaoka, Suita, Osaka 5650871, Japan. OI Yamada, Kenneth/0000-0003-1512-6805 NR 58 TC 88 Z9 92 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 MAY 31 PY 2002 VL 277 IS 22 BP 19922 EP 19928 DI 10.1074/jbc.M200383200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800090 PM 11891225 ER PT J AU Hang, J Dasso, M AF Hang, J Dasso, M TI Association of the human SUMO-1 protease SENP2 with the nuclear pore SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GTPASE-ACTIVATING PROTEIN; AXIN-BINDING PROTEIN; NUCLEOPORIN NUP153; COMPLEX PROTEIN; UBIQUITIN; IMPORT; RANGAP1; EXPORT; YEAST; LOCALIZATION AB SUMO-1 is a small ubiquitin-like protein that can be covalently conjugated to other proteins. A family of proteases catalyzes deconjugation of SUMO-1-containing species. Members of this family also process newly synthesized SUMO-1 into its conjugatable form. To understand these enzymes better, we have examined the localization and behavior of the human SUMO-1 protease SENP2. Here we have shown that SENP2 associates with the nuclear face of nuclear pores and that this association requires protein sequences near the N terminus of SENP2. We have also shown that SENP2 binds to Nup153, a nucleoporin that is localized to the nucleoplasmic face of the pore. Nup153 binding requires the same domain of SENP2 that mediates its targeting in vivo. Removal of the Nup153-interacting region of SENP2 results in a significant change in the spectrum of SUMO-1 conjugates within the cell. Our results suggest that association with the pore plays an important negative role in the regulation of SENP2, perhaps by restricting its activity to a subset of the conjugated proteins within the nucleus. C1 NICHHD, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA. RP Dasso, M (reprint author), NICHHD, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA. OI Dasso, Mary/0000-0002-5410-1371 FU NICHD NIH HHS [Z01 HD 01902-05] NR 28 TC 151 Z9 161 U1 1 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 31 PY 2002 VL 277 IS 22 BP 19961 EP 19966 DI 10.1074/jbc.M201799200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800095 PM 11896061 ER PT J AU Balla, A Tuymetova, G Barshishat, M Geiszt, M Balla, T AF Balla, A Tuymetova, G Barshishat, M Geiszt, M Balla, T TI Characterization of type II phosphatidylinositol 4-kinase isoforms reveals association of the enzymes with endosomal vesicular compartments SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PHOSPHOINOSITIDE KINASES; SACCHAROMYCES-CEREVISIAE; MEMBRANE TRAFFICKING; PROTEIN-KINASE; LIPID KINASE; YEAST; WORTMANNIN; CLATHRIN; GENE; REQUIREMENT AB Phosphorylation of phosphatidylinositol (PI) to PI 4-phosphate is one of the key reactions in the production of phosphoinositides, lipid regulators of several cellular functions. This reaction is catalyzed by multiple enzymes that belong either to the type II or the type III family of PI 4-kinases. Type III enzymes are structurally similar to PI 3-kinases and are sensitive to PI 3-kinase inhibitors. In contrast, the recent cloning of the first type II PI 4-kinase enzyme defined a novel enzyme family. Here we characterize a new member of this family, the type IIbeta enzyme that has been identified in the NCBI data base based on its homology to the first-cloned type IIalpha enzyme. The type IIbeta enzyme has a primary transcript size of similar to3.8 kb and shows wide tissue distribution. It contains an open reading frame of 1.4 kb, encoding a protein of similar to54 kDa. Sequence comparison reveals a high degree of similarity to the type IIalpha enzyme within the C-terminal catalytic domain but significantly lower homology within the N-terminal region. Expression of both enzyme yields increased PI 4-kinase activity that is associated with the microsomal membrane fractions and is significantly lower for the type IIbeta than the type IIalpha form. Both enzymes use PI as their primary substrate and have no detectable activity on PI monophosphates. Epitope-tagged as well as green fluorescent protein-tagged forms of both enzymes localize primarily to intracellular membranes and show prominent co-localization with early endosomes and recycling endosomes but not with the Golgi. These compartments participate in the processing of both the transferrin receptor and the G protein-coupled AT(1A) angiotensin receptor. Our data indicate the existence of multiple forms of type II PI 4-kinase in mammalian cells and suggest that their functions are related to the endocytic pathway. C1 Natl Inst Hlth, NIAID, Lab Host Def, Bethesda, MD 20892 USA. NICHHD, Endocrinol & Reprod Res Branch, Bethesda, MD 20892 USA. RP Balla, T (reprint author), Natl Inst Hlth, NIAID, Lab Host Def, Bldg 49 Rm 6A35 49 Convent Dr, Bethesda, MD 20892 USA. OI Balla, Tamas/0000-0002-9077-3335; Balla, Andras/0000-0002-6450-2793 NR 34 TC 111 Z9 114 U1 0 U2 8 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 MAY 31 PY 2002 VL 277 IS 22 BP 20041 EP 20050 DI 10.1074/jbc.M111807200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 557EH UT WOS:000175894800105 PM 11923287 ER PT J AU Tittor, J Paula, S Subramaniam, S Heberle, J Henderson, R Oesterhelt, D AF Tittor, J Paula, S Subramaniam, S Heberle, J Henderson, R Oesterhelt, D TI Proton translocation by bacteriorhodopsin in the absence of substantial conformational changes SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE retinal protein; photocycle; spectroscopy; vectorial catalysis ID DIFFERENCE SPECTROSCOPY; INFRARED-SPECTRA; RETINAL PROTEINS; SCHIFF-BASE; PHOTOCYCLE; INTERMEDIATE; MEMBRANE; FTIR; HALORHODOPSIN; RESOLUTION AB Unlike wild-type bacteriorhodopsin (BR), the BR triple mutant D96G/F171C/F219L has been shown to undergo only minor structural rearrangements during its photocycle. Nonetheless, the mutant is capable of transporting protons at a rate of 125(+/-40) H(+)/BR per minute under light-saturating conditions. Light adaptation of the triple mutant's retinal proceeds in a pH-dependent manner LIP to a maximum of 63%, all-trans. These two findings imply that the transport activity of the triple mutant comprises 66% of the wild-type activity. Time-resolved spectroscopy reveals that the identity and sequence of intermediates in the photocycle of the triple mutant in the all-trans configuration correspond to that of wild-type BR. The only differences relate to a slower rise and decay of the M and O intermediates, and a significant spectral contribution from a 13-cis component. No indication for accumulation of the N intermediate is found under a variety of conditions that normally favor the formation of this species in wild-type BR. The Fourer transform infrared (FTIR) spectrum of the M intermediate in the triple mutant resembles that of wild type. Minor changes in the amide I region during the photocycle suggest that only small movements of the protein backbone occur. Electron microscopy reveals large differences in conformation between the unilluminated state of the mutant protein and wild-type but no light-induced changes in time-resolved measurements. Evidently, proton transport by the triple mutant does not require the major conformational rearrangements that occur on the same time-scale with wild-type. Thus, we conclude that large conformational changes observed in the photocycle of the wild-type and many BR mutants are not a prerequisite for the change in accessibility of the Schiff base nitrogen atom that must occur during vectorial catalysis to allow, proton transport. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Max Planck Inst Biochem, D-82152 Martinsried, Germany. NCI, Bethesda, MD 20892 USA. Forschungszentrum Julich, IBI 2, D-52425 Julich, Germany. MRC, Mol Biol Lab, Cambridge CB2 2QH, England. RP Tittor, J (reprint author), Max Planck Inst Biochem, D-82152 Martinsried, Germany. EM tittor@biochem.mpg.de RI Heberle, Joachim/D-8605-2016 OI Heberle, Joachim/0000-0001-6321-2615 NR 41 TC 29 Z9 30 U1 1 U2 6 PU ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD MAY 31 PY 2002 VL 319 IS 2 BP 555 EP 565 DI 10.1016/S0022-2836(02)00307-8 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 562ZP UT WOS:000176229100025 PM 12051928 ER PT J AU Copeland, NG Jenkins, NA O'Brien, SJ AF Copeland, NG Jenkins, NA O'Brien, SJ TI Genomics: Mmu 16 - Comparative genomic highlights SO SCIENCE LA English DT Editorial Material ID PLACENTAL MAMMAL RADIATION; MOUSE C1 NCI, Mouse Canc Genet Program, Frederick, MD 21702 USA. NCI, Lab Genome Divers, Frederick, MD 21702 USA. RP Copeland, NG (reprint author), NCI, Mouse Canc Genet Program, Frederick, MD 21702 USA. NR 10 TC 10 Z9 10 U1 0 U2 0 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD MAY 31 PY 2002 VL 296 IS 5573 BP 1617 EP 1618 DI 10.1126/science.1073127 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 558PR UT WOS:000175976200028 PM 12040165 ER PT J AU Vaudry, D Stork, PJS Lazarovici, P Eiden, LE AF Vaudry, D Stork, PJS Lazarovici, P Eiden, LE TI Signaling pathways for PC12 cell differentiation: Making the right connections SO SCIENCE LA English DT Editorial Material ID NERVE GROWTH-FACTOR; NEURITE OUTGROWTH; REGULATED KINASE; RAS; TRANSCRIPTION; ACTIVATION; PROTEIN; FORM; CBP AB A key issue in signal transduction is how signaling pathways common to many systems-so-called canonical signaling cassettes-integrate signals from molecules having a wide spectrum of activities, such as hormones and neurotrophins, to deliver distinct biological outcomes. The neuroendocrine cell line PC12, derived from rat pheochromocytoma, provides an example of how one canonical signaling cassette-the Raf --> mitogen-activated protein kinase kinase (MEK) --> extracellular signal-regulated kinase (ERK) pathway-can promote distinct outcomes, which in this case include neuritogenesis, gene induction, and proliferation. Two growth hormones, epidermal growth factor (EGF) and nerve growth factor (NGF), use the same pathway to cause PC12 proliferation and differentiation, respectively. In addition, pituitary adenylate cyclase-activating polypeptide (PACAP), a neurotransmitter that, also causes differentiation, uses the same canonical cassette as NGF but in a different way. The Connections Map for PC12 Cell Differentiation brings into focus the complex array of specific cellular responses that rely on canonical signal transduction systems. C1 NIMH, Mol Neurosci Sect, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. Oregon Hlth Sci Univ, Vollum Inst, Portland, OR 97201 USA. Hebrew Univ Jerusalem, Sch Pharm, Dept Pharmacol, IL-91120 Jerusalem, Israel. RP Eiden, LE (reprint author), NIMH, Mol Neurosci Sect, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. OI Eiden, Lee/0000-0001-7524-944X NR 21 TC 540 Z9 550 U1 7 U2 64 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 MAY 31 PY 2002 VL 296 IS 5573 BP 1648 EP 1649 DI 10.1126/science.1071552 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 558PR UT WOS:000175976200044 PM 12040181 ER PT J AU Urizar, NL Liverman, AB Dodds, DT Silva, FV Ordentlich, P Yan, YZ Gonzalez, FJ Heyman, RA Mangelsdorf, DJ Moore, DD AF Urizar, NL Liverman, AB Dodds, DT Silva, FV Ordentlich, P Yan, YZ Gonzalez, FJ Heyman, RA Mangelsdorf, DJ Moore, DD TI A natural product that lowers cholesterol as an antagonist ligand for FXR SO SCIENCE LA English DT Article ID 7-ALPHA-HYDROXYLASE GENE CYP7A; ORPHAN NUCLEAR RECEPTOR; PREGNANE-X RECEPTOR; BILE-ACID; TARGETED DISRUPTION; COMMIPHORA MUKUL; PROTEIN GENE; IDENTIFICATION; ACTIVATION; MICE AB Extracts of the resin of the guggul tree (Commiphora mukul) tower LDL (low-density lipoprotein) cholesterol levels in humans. The plant sterol guggul-sterone [4,17(20)-pregnadiene-3,16-dione] is the active agent in this extract. We show that guggulsterone is a highly efficacious antagonist of the farnesoid X receptor (FXR), a nuclear hormone receptor that is activated by bile acids. Guggulsterone treatment decreases hepatic cholesterol in wild-type mice fed a high-cholesterol diet but is not effective in FXR-null mice. Thus, we propose that inhibition of FXR activation is the basis for the cholesterol-lowering activity of guggulsterone. Other natural products with specific biologic effects may modulate the activity of FXR or other relatively promiscuous nuclear hormone receptors. C1 Baylor Coll Med, Dept Mol & Cellular Biol, Houston, TX 77030 USA. Univ Texas, SW Med Ctr, Howard Hughes Med Inst, Dallas, TX 75390 USA. X Ceptor Therapeut Inc, San Diego, CA 92121 USA. NCI, Lab Metab, Ctr Canc Res, Bethesda, MD 20892 USA. RP Moore, DD (reprint author), Baylor Coll Med, Dept Mol & Cellular Biol, 1 Baylor Plaza, Houston, TX 77030 USA. NR 39 TC 296 Z9 323 U1 4 U2 28 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 MAY 31 PY 2002 VL 296 IS 5573 BP 1703 EP 1706 DI 10.1126/science.1072891 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 558PR UT WOS:000175976200064 PM 11988537 ER PT J AU Shidara, M Richmond, BJ AF Shidara, M Richmond, BJ TI Anterior cingulate: Single neuronal signals related to degree of reward expectancy SO SCIENCE LA English DT Article ID OBSESSIVE-COMPULSIVE DISORDER; RHESUS-MONKEY; SYMPTOM PROVOCATION; MOTOR AREAS; HUMAN BRAIN; CORTEX; ACTIVATION; BEHAVIOR; SELECTION; TOMOGRAPHY AB As monkeys perform schedules containing several trials with a visual cue indicating reward proximity, their error rates decrease as the number of remaining trials decreases, suggesting that their motivation and/or reward expectancy increases as the reward approaches. About one-third of single neurons recorded in the anterior cingulate cortex of monkeys during these reward schedules had responses that progressively changed strength with reward expectancy, an effect that disappeared when the cue was random. Alterations of this progression could be the basis for the changes from normal that are reported in anterior cingulate population activity for obsessive-compulsive disorder and drug abuse, conditions characterized by disturbances in reward expectancy. C1 Natl Inst Adv Ind Sci & Technol, Neurosci Res Inst, Tsukuba, Ibaraki 3058568, Japan. NIMH, Neuropsychol Lab, Bethesda, MD 20892 USA. RP Shidara, M (reprint author), Natl Inst Adv Ind Sci & Technol, Neurosci Res Inst, 1-1-1 Umezono, Tsukuba, Ibaraki 3058568, Japan. RI Marion-Poll, Frederic/D-8882-2011 OI Marion-Poll, Frederic/0000-0001-6824-0180 NR 39 TC 314 Z9 319 U1 2 U2 13 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 MAY 31 PY 2002 VL 296 IS 5573 BP 1709 EP 1711 DI 10.1126/science.1069504 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 558PR UT WOS:000175976200066 PM 12040201 ER PT J AU Rennels, MB Deloria, MA Pichichero, ME Englund, JA Anderson, EL Steinhoff, MC Decker, MD Edwards, KM AF Rennels, MB Deloria, MA Pichichero, ME Englund, JA Anderson, EL Steinhoff, MC Decker, MD Edwards, KM TI Lack of consistent relationship between quantity of aluminum in diphtheria-tetanus-acellular pertussis vaccines and rates of extensive swelling reactions SO VACCINE LA English DT Article; Proceedings Paper CT Workshop on Aluminum Adjuvants in Vaccines CY MAY 11-12, 2000 CL SAN JUAN, PUERTO RICO DE aluminum; vaccines; adsorbed ID WHOLE-CELL; ADVERSE REACTIONS; IMMUNOGENICITY; IMMUNIZATION; VACCINATION; CHILDREN AB The precise role that aluminum plays in local reactogenicity is not clear. We explored the relationship between rates of severe local reactions following the fourth and fifth booster doses of several diphtheria-tetanus-acellular pertussis vaccines (DTaP) and the quantity of aluminum contained in the different vaccines. Although there was a significant relationship between higher aluminum contents and swelling reactions >50 mm after dose 5, no relationship was seen with entire thigh swelling or with swelling >50 mm after dose 4. Because of the inconsistency of the data, a dose response between local reactogenicity and aluminum is questionable. (C) 2002 Published by Elsevier Science Ltd. C1 Univ Maryland, Sch Med, Baltimore, MD 21201 USA. NIH, Baltimore, MD 21201 USA. Univ Rochester, Sch Med, Rochester, NY 14642 USA. Univ Chicago, Chicago, IL 60637 USA. St Louis Univ, Sch Med, St Louis, MO 63110 USA. Johns Hopkins Sch Publ Hlth, Baltimore, MD 21201 USA. Vanderbilt Univ, Sch Med, Nashville, TN 37212 USA. RP Rennels, MB (reprint author), Univ Maryland, Sch Med, 22 S Greene St, Baltimore, MD 21201 USA. FU NIAID NIH HHS [N01-AI02645, N01-AI-62515, N01-AI05049, N01-AI05051, N01-AI15096, N01-AI72629] NR 15 TC 17 Z9 17 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 MAY 31 PY 2002 VL 20 SU 3 BP S44 EP S47 AR PII S0264-410X(02)00171-8 DI 10.1016/S0264-410X(02)00171-8 PG 4 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 568JY UT WOS:000176539700009 PM 12184364 ER PT J AU Boudsocq, F Ling, H Yang, W Woodgate, R AF Boudsocq, F Ling, H Yang, W Woodgate, R TI Structure-based interpretation of missense mutations in Y-family DNA polymerases and their implications for polymerase function and lesion bypass SO DNA REPAIR LA English DT Review DE DNA replication; pol IV; pol V; pol eta; pol iota; pol kappa; Rev 1 ID SULFOLOBUS-SOLFATARICUS P2; UMUD'C PROTEIN COMPLEX; ESCHERICHIA-COLI DINB; CRYSTAL-STRUCTURE; ERROR-PRONE; SACCHAROMYCES-CEREVISIAE; XERODERMA-PIGMENTOSUM; SALMONELLA-TYPHIMURIUM; INDUCED MUTAGENESIS; ULTRAVIOLET-LIGHT AB Our understanding of the molecular mechanisms of error-prone lesion bypass has changed dramatically in the past few years. The concept that the key participants in the mutagenic process were accessory proteins that somehow modified the ability of the cell's main replicase to facilitate bypass of normally blocking lesions has been replaced with one in which the replicase is displaced by a polymerase specialized in lesion bypass. The participants in this process remain the same, only their function has been reassigned. What was once known as the UmuC/DinB/Rev1/Rad30 superfamily of mutagenesis proteins, is now known as the Y-family of DNA polymerases. Quite remarkably, within the space of 3 years, the field has advanced from the initial discovery of intrinsic polymerase function, to the determination of the tertiary structures of several Y-family DNA polymerases. A key to determining the biochemical properties of each DNA polymerase is through structure-function studies that result in the site-specific substitution of particular amino acids at critical sites within each DNA polymerase. However, we should not forget the power of genetic selection that allows us to identify residues within each polymerase that are generated by "random mutagenesis" and which are important for both a gain or loss of function in vivo. In this review, we discuss the structural ramifications of several missense mutations previously identified in various Y-family DNA polymerase and speculate on how each amino acid substitution might modify the enzymatic activity of the respective polymerase or possibly perturb protein-protein interactions necessary for efficient translesion replication in vivo. Published by Elsevier Science B.V. C1 NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bethesda, MD 20892 USA. NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Woodgate, R (reprint author), NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bldg 6,Room 1A13,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Ling, Hong/E-3729-2010; Yang, Wei/D-4926-2011 OI Yang, Wei/0000-0002-3591-2195 NR 102 TC 66 Z9 67 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1568-7864 J9 DNA REPAIR JI DNA Repair PD MAY 30 PY 2002 VL 1 IS 5 BP 343 EP 358 AR PII S1568-7864(02)00019-8 DI 10.1016/S1568-7864(02)00019-8 PG 16 WC Genetics & Heredity; Toxicology SC Genetics & Heredity; Toxicology GA 597YM UT WOS:000178248400002 PM 12509239 ER PT J AU Okazawa, H Rich, T Chang, A Lin, X Waragai, M Kajikawa, M Enokido, Y Komuro, A Kato, S Shibata, M Hatanaka, H Mouradian, MM Sudol, M Kanazawa, I AF Okazawa, H Rich, T Chang, A Lin, X Waragai, M Kajikawa, M Enokido, Y Komuro, A Kato, S Shibata, M Hatanaka, H Mouradian, MM Sudol, M Kanazawa, I TI Interaction between mutant ataxin-1 and POBP-1 affects transcription and cell death SO NEURON LA English DT Article ID RNA-POLYMERASE-II; NEURONAL INTRANUCLEAR INCLUSIONS; EXPANDED POLYGLUTAMINE PROTEIN; CARBOXYL-TERMINAL DOMAIN; CREB-BINDING PROTEIN; SPINOCEREBELLAR ATAXIA; HUNTINGTONS-DISEASE; NUCLEAR INCLUSIONS; TRACT-BINDING; WW DOMAIN AB PQBP-1 was isolated on the basis of its interaction with polyglutamine tracts. In this study, using in vitro and in vivo assays, we show that the association between ataxin-1 and PQBP-1 is positively influenced by expanded polyglutamine sequences. In cell lines, interaction between the two molecules induces apoptotic cell death. As a possible mechanism underlying this phenomenon, we found that mutant ataxin-1 enhances binding of PQBP-1 to the C-terminal domain of RNA polymerase II large subunit (Pol II). This reduces the level of phosphorylated Pol II and transcription. Our results suggest the involvement of PQBP-1 in the pathology of spinocerebellar ataxia type 1 (SCA1) and support the idea that modified transcription underlies polyglutamine-mediated pathology. C1 Univ Tokyo, Grad Sch med, Dept Neurol, Bunkyo Ku, Tokyo 1138655, Japan. MBL Co Ltd, Dept Pharmaceut Dev, Nagano 3960002, Japan. Osaka Univ, Inst Prot Res, Div Prot Biosynth, Suita, Osaka 5650871, Japan. JST, ERATO, Sagami Chem Res Ctr, Kanagawa 2290012, Japan. Univ Cambridge, Dept Pathol, Cambridge CB2 1QP, England. Mt Sinai Med Ctr, Dept Med, New York, NY 10029 USA. Baylor Coll Med, Howard Hughes Med Inst, Dept Pediat Neurol Neurosci & Mol Genet, Houston, TX 77030 USA. NINCDS, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Okazawa, H (reprint author), Tokyo Metropolitan Inst Neurosci, Dept Mol Therapeut, 2-6 Musashidai, Tokyo 1838526, Japan. NR 56 TC 124 Z9 128 U1 0 U2 2 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTS AVE,, CAMBRIDGE, MA 02138 USA SN 0896-6273 J9 NEURON JI Neuron PD MAY 30 PY 2002 VL 34 IS 5 BP 701 EP 713 DI 10.1016/S0896-6273(02)00697-9 PG 13 WC Neurosciences SC Neurosciences & Neurology GA 558DV UT WOS:000175951800006 PM 12062018 ER PT J AU Skyler, JS Brown, D Chase, HP Collier, E Cowie, C Eisenbarth, GS Fradkin, J Grave, G Greenbaum, C Jackson, RA Kaufman, FR Krischer, JP Marks, JB Palmer, JP Ricker, A Schatz, DA Wilson, D Winter, WE Wolfsdorf, J Zeidler, A Dickler, H Eastman, RC Maclaren, NK Malone, JI Robertson, PR Skyler, JS Krischer, JP Wolfsdorf, J Cowie, C Palmer, JP Greenbaum, C Cuthbertson, D Rafkin-Mervis, LM Kaufman, FR Chase, HP Palmer, JP Chase, HP Cowie, C Fradkin, J Eisenbarth, GS Greenbaum, C Herold, K Kaufman, FR Krischer, JP Marks, JB Rafkin-Mervis, L Schatz, DA Skyler, JS Aneju, B Conboy, D Cook, R Dennis, MA Finney, L Harris, S Matheson, D McCulloch-Olsen, M Smith, T Valenzuela, J Vega, N Crofford, OB DeMets, D Lachin, JM Nerup, J Rossini, A Schiffrin, A Steffes, M Tsiatis, A Zinman, B AF Skyler, JS Brown, D Chase, HP Collier, E Cowie, C Eisenbarth, GS Fradkin, J Grave, G Greenbaum, C Jackson, RA Kaufman, FR Krischer, JP Marks, JB Palmer, JP Ricker, A Schatz, DA Wilson, D Winter, WE Wolfsdorf, J Zeidler, A Dickler, H Eastman, RC Maclaren, NK Malone, JI Robertson, PR Skyler, JS Krischer, JP Wolfsdorf, J Cowie, C Palmer, JP Greenbaum, C Cuthbertson, D Rafkin-Mervis, LM Kaufman, FR Chase, HP Palmer, JP Chase, HP Cowie, C Fradkin, J Eisenbarth, GS Greenbaum, C Herold, K Kaufman, FR Krischer, JP Marks, JB Rafkin-Mervis, L Schatz, DA Skyler, JS Aneju, B Conboy, D Cook, R Dennis, MA Finney, L Harris, S Matheson, D McCulloch-Olsen, M Smith, T Valenzuela, J Vega, N Crofford, OB DeMets, D Lachin, JM Nerup, J Rossini, A Schiffrin, A Steffes, M Tsiatis, A Zinman, B CA Diabetes Prevention Trial-Type 1 D TI Effects of insulin in relatives of patients with type 1 diabetes mellitus SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article; Proceedings Paper CT 61st Annual Meeting of the American-Diabetes-Association CY JUN 22-26, 2001 CL PHILADELPHIA, PA SP Amer Diabet Assoc ID BB WOR RATS; ISLET-CELL; HIGH-RISK; POSITIVE RELATIVES; ADOPTIVE TRANSFER; NOD MICE; IDDM; AUTOANTIBODY; PREVENTION; PROPHYLAXIS AB Background: It is unknown whether insulin therapy can delay or prevent diabetes in nondiabetic relatives of patients with diabetes. Methods: In a randomized, controlled, nonblinded clinical trial, we screened 84,228 first-degree and second-degree relatives of patients with diabetes for islet-cell antibodies; 3152 tested positive; 2103 of the 3152 underwent genetic, immunologic, and metabolic staging to quantify their risk; 372 of the 2103 had a projected five-year risk of more than 50 percent; 339 of the 372 (median age, 11.2 years) were randomly assigned to undergo either close observation or an intervention that consisted of low-dose subcutaneous ultralente insulin, administered twice daily for a total dose of 0.25 unit per kilogram of body weight per day, plus annual four-day continuous intravenous infusions of insulin. Oral glucose-tolerance tests were performed every six months. Median follow-up was 3.7 years. The primary end point was a diagnosis of diabetes. Results: Diabetes was diagnosed in 69 subjects in the intervention group and 70 subjects in the observation group. The annualized rate of progression to diabetes was 15.1 percent in the intervention group and 14.6 percent in the observation group. The cumulative incidence of diabetes was similar in the two groups (relative risk in the intervention group as compared with the observation group, 0.96). Most subjects in whom diabetes developed were asymptomatic. Progression to diabetes occurred at a faster rate among subjects with abnormal base-line glucose tolerance (22 percent per year) than among those with normal base-line glucose tolerance (10 percent per year, P<0.001). There were no episodes of severe hypoglycemia. The incidence of chemical hypoglycemia, assessed without ascertainment bias, was similar in the two groups. Conclusions: In persons at high risk for diabetes, insulin at the dosage used in this study does not delay or prevent type 1 diabetes. C1 Univ Miami, Miami, FL 33101 USA. Univ Minnesota, Minneapolis, MN 55455 USA. Univ Colorado, Boulder, CO 80309 USA. NIDDK, Bethesda, MD 20892 USA. Childrens Hosp Los Angeles, Los Angeles, CA USA. Univ S Florida, Tampa, FL 33620 USA. Univ Miami, Coral Gables, FL 33124 USA. Univ Washington, Seattle, WA 98195 USA. Childrens Hosp, Boston, MA 02115 USA. Univ Florida, Gainesville, FL 32611 USA. Stanford Univ, Stanford, CA 94305 USA. Univ So Calif, Los Angeles, CA 90089 USA. Columbia Univ, New York, NY 10027 USA. Univ Wisconsin, Madison, WI 53706 USA. George Washington Univ, Washington, DC 20052 USA. Univ Copenhagen, DK-1168 Copenhagen, Denmark. Univ Massachusetts, Amherst, MA 01003 USA. McGill Univ, Montreal, PQ H3A 2T5, Canada. N Carolina State Univ, Raleigh, NC 27695 USA. Univ Toronto, Toronto, ON, Canada. RP Skyler, JS (reprint author), Univ Miami, POB 016960, Miami, FL 33101 USA. EM jskyler@miami.edu RI Skyler, Jay/F-4211-2016; OI Lachin, John/0000-0001-9838-2841 NR 31 TC 422 Z9 434 U1 4 U2 15 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 MAY 30 PY 2002 VL 346 IS 22 BP 1685 EP 1691B PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 556DM UT WOS:000175834100002 ER PT J AU Herold, KC Hagopian, W Auger, JA Poumian-Ruiz, E Taylor, L Donaldson, D Gitelman, SE Harlan, DM Xu, DL Zivin, RA Bluestone, JA AF Herold, KC Hagopian, W Auger, JA Poumian-Ruiz, E Taylor, L Donaldson, D Gitelman, SE Harlan, DM Xu, DL Zivin, RA Bluestone, JA TI Anti-CD3 monoclonal antibody in new-onset type 1 diabetes mellitus SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID T-CELLS; OKT3 ANTIBODY; RECEPTOR; CYCLOSPORINE; REMISSION; MICE; IMMUNOSUPPRESSION; INDUCTION; SUBSETS; DISEASE AB Background: Type 1 diabetes mellitus is a chronic autoimmune disease caused by the pathogenic action of T lymphocytes on insulin-producing beta cells. Previous clinical studies have shown that continuous immune suppression temporarily slows the loss of insulin production. Preclinical studies suggested that a monoclonal antibody against CD3 could reverse hyperglycemia at presentation and induce tolerance to recurrent disease. Methods: We studied the effects of a nonactivating humanized monoclonal antibody against CD3 - hOKT3gamma1(Ala-Ala) - on the loss of insulin production in patients with type 1 diabetes mellitus. Within 6 weeks after diagnosis, 24 patients were randomly assigned to receive either a single 14-day course of treatment with the monoclonal antibody or no antibody and were studied during the first year of disease. Results: Treatment with the monoclonal antibody maintained or improved insulin production after one year in 9 of the 12 patients in the treatment group, whereas only 2 of the 12 controls had a sustained response (P=0.01). The treatment effect on insulin responses lasted for at least 12 months after diagnosis. Glycosylated hemoglobin levels and insulin doses were also reduced in the monoclonal-antibody group. No severe side effects occurred, and the most common side effects were fever, rash, and anemia. Clinical responses were associated with a change in the ratio of CD4+ T cells to CD8+ T cells 30 and 90 days after treatment. Conclusions: Treatment with hOKT3gamma1(Ala-Ala) mitigates the deterioration in insulin production and improves metabolic control during the first year of type 1 diabetes mellitus in the majority of patients. The mechanism of action of the anti-CD3 monoclonal antibody may involve direct effects on pathogenic T cells, the induction of populations of regulatory cells, or both. C1 Columbia Univ, Coll Phys & Surg, Naomi Berrie Diabet Ctr, New York, NY 10032 USA. Columbia Univ, Coll Phys & Surg, Dept Med, Div Endocrinol, New York, NY USA. Pacific NW Res Inst, Seattle, WA USA. Univ Chicago, Chicago, IL 60637 USA. Univ Utah, Salt Lake City, UT USA. Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Med, San Francisco, CA USA. NIDDKD, Bethesda, MD 20892 USA. RW Johnson Pharmaceut Res Inst, Raritan, NJ 08869 USA. RP Herold, KC (reprint author), Columbia Univ, Coll Phys & Surg, Naomi Berrie Diabet Ctr, 1150 St Nicholas Ave, New York, NY 10032 USA. EM kh318@columbia.edu FU NCRR NIH HHS [M01 RR00645, M01 RR01271]; NIDDK NIH HHS [P60 DK20595, R01DK57846]; PHS HHS [U19A146132] NR 40 TC 711 Z9 736 U1 4 U2 19 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 EI 1533-4406 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 30 PY 2002 VL 346 IS 22 BP 1692 EP 1698 DI 10.1056/NEJMoa012864 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 556DM UT WOS:000175834100003 PM 12037148 ER PT J AU Walsh, TJ Pappas, PG Winston, DJ AF Walsh, TJ Pappas, PG Winston, DJ TI Voriconazole versus liposomal amphotericin B for empirical antifungal therapy - Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter ID NEUTROPENIC PATIENTS; PERSISTENT FEVER; CANCER; TRIAL C1 NCI, Bethesda, MD 20892 USA. Univ Alabama, Birmingham, AL 35294 USA. Univ Calif Los Angeles, Los Angeles, CA USA. RP Walsh, TJ (reprint author), NCI, Bethesda, MD 20892 USA. NR 4 TC 3 Z9 3 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 30 PY 2002 VL 346 IS 22 BP 1746 EP 1747 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 556DM UT WOS:000175834100015 ER PT J AU Dalakas, MC AF Dalakas, MC TI High-dose intravenous immune globulin for Stiff-Person syndrome - Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter ID IMMUNOGLOBULIN; AUTOIMMUNE; DISEASES C1 NIH, Bethesda, MD 20892 USA. RP Dalakas, MC (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 4 TC 0 Z9 0 U1 0 U2 1 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 30 PY 2002 VL 346 IS 22 BP 1747 EP 1748 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 556DM UT WOS:000175834100018 ER PT J AU Nutman, TB AF Nutman, TB TI Case 1-2002: Loa loa - Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Nutman, TB (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 4 TC 0 Z9 0 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 30 PY 2002 VL 346 IS 22 BP 1751 EP 1751 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 556DM UT WOS:000175834100025 ER PT J AU Becker, LB Weisfeldt, ML Weil, MH Budinger, T Carrico, J Kern, K Nichol, G Shechter, I Traystman, R Webb, C Wiedemann, H Wise, R Sopko, G AF Becker, LB Weisfeldt, ML Weil, MH Budinger, T Carrico, J Kern, K Nichol, G Shechter, I Traystman, R Webb, C Wiedemann, H Wise, R Sopko, G TI The PULSE initiative - Scientific priorities and strategic planning for resuscitation research and life saving therapies SO CIRCULATION LA English DT Article ID MYOCARDIAL REPERFUSION INJURY; TRANSIENT CEREBRAL-ISCHEMIA; CARDIAC-ARREST; CARDIOPULMONARY-RESUSCITATION; MILD HYPOTHERMIA; BODY-TEMPERATURE; BLOOD-FLOW; DELAYED RESUSCITATION; MODERATE HYPOTHERMIA; SUSPENDED ANIMATION C1 Univ Chicago, Dept Med, Chicago, IL 60637 USA. Inst Crit Care Med, Palm Springs, CA USA. Univ Calif Berkeley, Berkeley, CA 94720 USA. Univ Texas SW, Dallas, TX USA. Univ Arizona, Tucson, AZ USA. Univ Ottawa, Ottawa, ON, Canada. Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA. Johns Hopkins Univ, Baltimore, MD USA. Cleveland Clin Fdn, Cleveland, OH 44195 USA. NHLBI, Bethesda, MD 20892 USA. RP Becker, LB (reprint author), Univ Chicago, Dept Med, MC5068,5841 S Maryland Ave, Chicago, IL 60637 USA. RI Crouser, Elliott/E-2933-2011; OI Wiedemann, Herbert/0000-0002-4587-4401; Wise, Robert/0000-0002-8353-2349 NR 86 TC 40 Z9 45 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 MAY 28 PY 2002 VL 105 IS 21 BP 2562 EP 2570 DI 10.1161/01.CIR.0000017142.3991.C3 PG 9 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 557UX UT WOS:000175927500027 PM 12034666 ER PT J AU Borkowf, CB AF Borkowf, CB TI Computing the nonnull asymptotic variance and the asymptotic relative efficiency of Spearman's rank correlation SO COMPUTATIONAL STATISTICS & DATA ANALYSIS LA English DT Article DE asymptotic relative efficiency; asymptotic variance; empirical distribution function; Fisher's z-transformation; nonparametric simulation; Spearman's rank correlation ID DISTRIBUTIONS AB Over the past century, Spearman's rank correlation, rho(s), has become one of the most commonly used nonparametric statistics, yet much remains unknown about its finite and asymptotic behavior. This paper presents a method for computing the asymptotic variance of the point estimate, (ρ) over cap (s), in terms of expectations of the joint and marginal distribution functions, for any underlying bivariate distribution that satisfies minimal regularity conditions. Also presented are numerical results for certain bivariate distributions of interest in order to demonstrate that distributions with similar values of Pearson's or Spearman's correlations can yield surprisingly different values for the asymptotic variance of (ρ) over cap (s). In particular, these results emphasize that one should not use certain standard procedures for hypothesis testing and confidence interval construction that assume bivariate normality without first checking this distributional assumption. Finally, these numerical results are used to compute the asymptotic relative efficiency of Spearman's rank correlation compared to Pearson's correlation. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NCI, Ctr Canc Res, Canc Prevent Studies Branch, Bethesda, MD 20892 USA. NR 16 TC 16 Z9 16 U1 3 U2 7 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-9473 J9 COMPUT STAT DATA AN JI Comput. Stat. Data Anal. PD MAY 28 PY 2002 VL 39 IS 3 BP 271 EP 286 AR PII S 0167-9473(01)00081-0 DI 10.1016/S0167-9473(01)00081-0 PG 16 WC Computer Science, Interdisciplinary Applications; Statistics & Probability SC Computer Science; Mathematics GA 561HG UT WOS:000176133700002 ER PT J AU Konkel, JT Fan, JF Jayachandran, B Kirk, KL AF Konkel, JT Fan, JF Jayachandran, B Kirk, KL TI Syntheses of 6-fluoro-meta-tyrosine and of its metabolites SO JOURNAL OF FLUORINE CHEMISTRY LA English DT Article DE fluorinated metabolites; PET scanning agents; HPLC standards ID TYROSINE AB 6-Fluoro-meta-tyrosine (1) was prepared from 2-fluoro-5-hydroxybenzaldehyde (6) based on an Erlenmeyer-Plochl azlactone strategy. Products of expected metabolism of the amino acid, including 6-fluoro-meta-tyramine (2) and its O-sulfate conjugate (3), (2-fluoro-5-hydrxoyphenyl)acetic acid (4), and 6-fluoro-meta-octopamine (5) also were prepared from 1. The use of a recently reported ultrasound-catalyzed Henry reaction facilitated the preparation of the tyramine derivative 2. The compounds synthesized are available for high performance liquid chromatography (HPLC) standards in positron emission tomography (PET) studies employing 6-[F-18]fluoro-meta-tyrosine and as reference samples for metabolic studies of the amino acid. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Kirk, KL (reprint author), NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NR 9 TC 21 Z9 21 U1 1 U2 3 PU ELSEVIER SCIENCE SA PI LAUSANNE PA PO BOX 564, 1001 LAUSANNE, SWITZERLAND SN 0022-1139 J9 J FLUORINE CHEM JI J. Fluor. Chem. PD MAY 28 PY 2002 VL 115 IS 1 BP 27 EP 32 AR PII S0022-1139(02)00013-1 DI 10.1016/S0022-1139(02)00013-1 PG 6 WC Chemistry, Inorganic & Nuclear; Chemistry, Organic SC Chemistry GA 564LR UT WOS:000176316300005 ER PT J AU Corboy, JR Boudreau, E Morgenlander, JC Rudnicki, S Coyle, PK AF Corboy, JR Boudreau, E Morgenlander, JC Rudnicki, S Coyle, PK TI Neurology residency training at the millennium SO NEUROLOGY LA English DT Article ID WORKFORCE C1 SUNY Stony Brook, Hlth Sci Ctr, Dept Neurol, Stony Brook, NY USA. Univ Colorado, Hlth Sci Ctr, Dept Neurol, Denver, CO 80202 USA. NINCDS, Epilepsy Res Branch, NIH, Bethesda, MD USA. Duke Univ, Med Ctr, Dept Med, Div Neurol, Durham, NC 27710 USA. Univ Arkansas, Dept Neurol, Little Rock, AR 72204 USA. RP Corboy, JR (reprint author), Amer Acad Neurol, 1080 Montreal Ave, St Paul, MN 55116 USA. NR 8 TC 16 Z9 17 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 MAY 28 PY 2002 VL 58 IS 10 BP 1454 EP 1460 PG 7 WC Clinical Neurology SC Neurosciences & Neurology GA 555LC UT WOS:000175794700003 PM 12034779 ER PT J AU Pasalar, P Najmabadi, H Noorian, AR Moghimi, B Jannati, A Soltanzadeh, A Krefft, T Crook, R Hardy, J AF Pasalar, P Najmabadi, H Noorian, AR Moghimi, B Jannati, A Soltanzadeh, A Krefft, T Crook, R Hardy, J TI An Iranian family with Alzheimer's disease caused by a novel APP mutation (Thr714Ala) SO NEUROLOGY LA English DT Editorial Material ID PRECURSOR PROTEIN GENE C1 NIA, Neurogenet Lab, NIH, Bethesda, MD 20817 USA. Univ Tehran Med Sci, Dept Biochem, Tehran, Iran. Univ Tehran Med Sci, Sch Med, Tehran, Iran. Univ Social Welfare & Rehabil, Dept Genet, Tehran, Iran. Univ Tehran Med Sci, Shariati Hosp, Dept Neurol, Tehran, Iran. Mayo Clin, Dept Neurol, Jacksonville, FL 32224 USA. Mayo Clin, Dept Neurosci, Jacksonville, FL 32224 USA. RP Hardy, J (reprint author), NIA, Neurogenet Lab, NIH, Bldg 10,6C, Bethesda, MD 20817 USA. EM hardyj@mail.nih.gov RI Hardy, John/C-2451-2009; Jannati, Ali/J-6453-2012; OI Najmabadi, Hossein/0000-0002-6084-7778; pasalar, parvin/0000-0002-8556-6729 NR 6 TC 28 Z9 30 U1 0 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD MAY 28 PY 2002 VL 58 IS 10 BP 1574 EP 1575 PG 2 WC Clinical Neurology SC Neurosciences & Neurology GA 555LC UT WOS:000175794700033 PM 12034808 ER PT J AU Alesci, S Ramsey, WJ Bornstein, SR Chrousos, GP Hornsby, PJ Benvenga, S Trimarchi, F Ehrhart-Bornstein, M AF Alesci, S Ramsey, WJ Bornstein, SR Chrousos, GP Hornsby, PJ Benvenga, S Trimarchi, F Ehrhart-Bornstein, M TI Adenoviral vectors can impair adrenocortical steroidogenesis: Clinical implications for natural infections and gene therapy SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID RECOMBINANT ADENOVIRUS; MEDIATED TRANSFER; CYSTIC-FIBROSIS; EXPRESSION; CANCER; CELLS; APOPTOSIS; TOXICITY; STRESS; SYSTEM AB Recombinant adenoviral vectors are effective in transferring foreign genes to a variety of cells and tissue types, both in vitro and in vivo. However, during the gene transfer, they may alter the principal function and local environment of transfected cells. Increasing evidence exists for a selective adrenotropism of adenovirus during infections and gene transfer. Therefore, using bovine adrenocortical cells in primary culture, we analyzed the influence of different adenoviral deletion mutants on cell morphology and physiology. Transfection of cells with an E1/E3-deleted adenoviral vector, engineered to express a modified form of the Aequorea victoria green fluorescent protein, was highly efficient, as documented by fluorescent microscopy. Ultrastructural analysis, however, demonstrated nuclear fragmentation and mitochondrial alterations in addition to intranuclear viral particles. Basal secretion of 17-OH-progesterone, 11-deoxycortisol, and cortisol was significantly increased by E1/E3-deleted vectors; yet, the corticotropin-stimulated release of these steroids was decreased. Interestingly, neither purified viral capsids nor E3/E4-deleted adenoviral mutants altered basal and stimulated steroidogenesis of adrenocortical cells. An intact adrenal response is crucial for adaptation to stress and survival. Therefore, the implications of our findings need to be considered in patients with adenoviral infections and those undergoing clinical studies using adenoviral gene transfer. At the same time, the high level of transfection in adrenocortical cells might make appropriately modified adenoviral vectors suitable for gene therapy of adrenocortical carcinomas with poor prognosis. C1 Natl Inst Child Hlth & Human Dev, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. Univ Messina, Sch Med, Dipartimento Clin Sperimentale Med & Farma, I-98125 Messina, Italy. Natl Human Genome Res Inst, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA. Univ Dusseldorf, Dept Endocrinol, D-40225 Dusseldorf, Germany. Univ Dusseldorf, German Diabetes Res Inst, D-40225 Dusseldorf, Germany. Univ Texas, Hlth Sci Ctr, Sam & Ann Barshop Ctr Longev & Aging Studies, San Antonio, TX 78245 USA. RP Alesci, S (reprint author), Natl Inst Child Hlth & Human Dev, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NR 41 TC 28 Z9 32 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 MAY 28 PY 2002 VL 99 IS 11 BP 7484 EP 7489 DI 10.1073/pnas.062170099 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 557KW UT WOS:000175908600031 PM 12032309 ER PT J AU Hoe, NP Ireland, RM DeLeo, FR Gowen, BB Dorward, DW Voyich, JM Liu, M Burns, EH Culnan, DM Bretscher, A Musser, JM AF Hoe, NP Ireland, RM DeLeo, FR Gowen, BB Dorward, DW Voyich, JM Liu, M Burns, EH Culnan, DM Bretscher, A Musser, JM TI Insight into the molecular basis of pathogen abundance: Group A Streptococcus inhibitor of complement inhibits bacterial adherence and internalization into human cells SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE microbiology; serotype M1; epidemic waves; ezrin ID GROUP-A STREPTOCOCCUS; HYALURONIC-ACID CAPSULE; ACTIN-BINDING-SITE; EPITHELIAL-CELLS; PROTEIN FAMILY; ERM PROTEINS; IN-VIVO; EZRIN; MOESIN; PYOGENES AB Streptococcal inhibitor of complement (Sic) is a secreted protein made predominantly by serotype M1 Group A Streptococcus (GAS), which contributes to persistence in the mammalian upper respiratory tract and epidemics of human disease. Unexpectedly, an isogenic sic-negative mutant adhered to human epithelial cells significantly better than the wild-type parental strain. Purified Sic inhibited the adherence of a sic negative serotype M1 mutant and of non-Sic-producing GAS strains to human epithelial cells. Sic was rapidly internalized by human epithelial cells, inducing cell flattening and loss of microvilli. Ezrin and moesin, human proteins that functionally link the cytoskeleton to the plasma membrane, were identified as Sic-binding proteins by affinity chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis. Sic colocalized with ezrin inside epithelial cells and bound to the F-actin-binding site region located in the carboxyl terminus of ezrin and moesin. Synthetic peptides corresponding to two regions of Sic had GAS adherence-inhibitory activity equivalent to mature Sic and inhibited binding of Sic to ezrin. In addition, the sic mutant was phagocytosed and killed by human polymorphonuclear leukocytes significantly better than the wild-type strain, and Sic colocalized with ezrin in discrete regions of polymorphonuclear leukocytes. The data suggest that binding of Sic to ezrin alters cellular processes critical for efficient GAS contact, internalization, and killing. Sic enhances bacterial survival by enabling the pathogen to avoid the intracellular environment. This process contributes to the abundance of M1 GAS in human infections and their ability to cause epidemics. C1 NIAID, Rocky Mt Lab, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. NIAID, Rocky Mt Lab, Microscopy Branch, NIH, Hamilton, MT 59840 USA. Drew Univ, Dept Biol, Madison, NJ 07940 USA. Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY 14853 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Lab, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. OI DeLeo, Frank/0000-0003-3150-2516 NR 30 TC 54 Z9 55 U1 1 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 MAY 28 PY 2002 VL 99 IS 11 BP 7646 EP 7651 DI 10.1097/pnas.112039899 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 557KW UT WOS:000175908600059 PM 12032337 ER PT J AU Iezzi, S Cossu, G Nervi, C Sartorelli, V Puri, PL AF Iezzi, S Cossu, G Nervi, C Sartorelli, V Puri, PL TI Stage-specific modulation of skeletal myogenesis by inhibitors of nuclear deacetylases SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE deacetylase inhibitors; gene expression; MyoD; skeletal muscle differentiation; embryos ID MUSCLE GENE-EXPRESSION; HISTONE DEACETYLASE; MOUSE EMBRYOGENESIS; CHROMATIN STRUCTURE; REGULATORY FACTORS; SODIUM-BUTYRATE; CELL-CYCLE; MYOD; DIFFERENTIATION; ACETYLATION AB Nuclear acetyltransferases promote and deacetylases inhibit skeletal muscle-gene expression, suggesting the potential effectiveness of deacetylase inhibitors (DIs) in modulating skeletal myogenesis. Surprisingly, previous studies have indicated that DIs suppress myogenesis. The recent observations that histone deacetylases associate with the muscle-regulatory proteins MyoD and MEF2C only in undifferentiated myoblasts prompted us to evaluate the effect of DIs at distinct stages of the myogenic program. We found that exposure of established rodent and human muscle cells to distinct DIs has stage-specific effects. Exposure of undifferentiated skeletal myoblasts to DIs, followed by incubation in differentiation medium, enhanced the expression of muscle-specific reporters and increased the levels of endogenous muscle proteins, leading to a dramatic increase in the formation of multinucleated myotubes. By contrast, simultaneous exposure of muscle cells to differentiation medium and DIs inhibited the myogenic program. Likewise, embryos exposed in utero to nonteratogenic doses of DI at the early stages of somitic myogenesis (embryonic day 8.5) exhibited an increased number of somites and augmented expression of a muscle-specific transgene as well as endogenous muscle genes. The functional effects induced by DIs were mirrored by changes in the state of acetylation of histones present at a muscle-gene enhancer and of MyoD itself. These results represent the first evidence that DIs can enhance muscle differentiation and suggest the rationale for their use in manipulating adult and embryonic skeletal myogenesis. C1 NIAMSD, Muscle Biol Lab, Muscle Gene Express Grp, NIH, Bethesda, MD 20892 USA. H San Raffaele, Inst Stem Cell Res, I-20133 Milan, Italy. Univ Roma La Sapienza, Dept Histol & Med Embryol, I-00161 Rome, Italy. Univ Roma La Sapienza, Lab Gene Express Fdn Andrea Cesalpino, I-00161 Rome, Italy. Salk Inst Biol Studies, Clayton Fdn Labs Peptide Biol, La Jolla, CA 92093 USA. RP Sartorelli, V (reprint author), NIAMSD, Muscle Biol Lab, Muscle Gene Express Grp, NIH, Bethesda, MD 20892 USA. OI NERVI, Clara/0000-0001-9341-0188; Puri, Pier Lorenzo/0000-0003-4964-0095 FU Telethon [TCP00081] NR 24 TC 58 Z9 58 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 MAY 28 PY 2002 VL 99 IS 11 BP 7757 EP 7762 DI 10.1073/pnas.112218599 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 557KW UT WOS:000175908600078 PM 12032356 ER PT J AU Weetall, M Digan, ME Hugo, R Mathew, S Hopf, C Tart-Risher, N Zhang, J Shi, V Fu, FM Hammond-McKibben, D West, S Brack, R Brinkmann, V Bergman, R Neville, D Lake, P AF Weetall, M Digan, ME Hugo, R Mathew, S Hopf, C Tart-Risher, N Zhang, J Shi, V Fu, FM Hammond-McKibben, D West, S Brack, R Brinkmann, V Bergman, R Neville, D Lake, P TI T-cell depletion and graft survival induced by anti-human CD3 immunotoxins in human CD3 epsilon transgenic mice SO TRANSPLANTATION LA English DT Article ID SINGLE-CHAIN IMMUNOTOXIN; IN-VIVO EFFICACY; MONOCLONAL-ANTIBODIES; DIPHTHERIA-TOXIN; RHESUS-MONKEYS; TOLERANCE; LYMPHOCYTES; ALLOGRAFTS; INDUCTION; TOXICITY AB Background. Amti-CD3 immunotoxins are broad-spectrum immunosuppressive agents in a wide range of organ transplantation animal models with potential use in eliciting antigen-specific tolerance. However, the anti-CD3 immunotoxins used in animal studies do not cross-react with human T cells, limiting extrapolation to humans and hindering clinical development. Methods. Three anti-human CD3-directed immunotoxins, DT389-scFv(UCHT1), scFv(UCHT1)-PE38, and UCHT1-CRM9, were compared in vitro and in transgenic mice, tgepsilon600+/-, that have T cells expressing both human and murine CD34epsilon antigens. Results. These immunotoxins were extraordinarily potent in vitro against human or transgenic mouse T cells, with IC50 values in cellular assays ranging from pM to fM. Systemic administration of these immunotoxins dose-dependently depleted >99% of tgepsilon600+/- lymph node and spleen T cells in vivo. Depletion was specific for T cells. The loss of the concanavalin A-induced, but not the lipopolysaccharide-induced, splenic proliferative response from immunotoxin-treated animals further demonstrated specific loss of T-cell function. Immunotoxin treatment prolonged fully allogeneic skin graft survival in tgepsilon600+/- recipients to 25 days from 10 days in untreated animals. T-cells recovered to similar to50% of normal levels after approximately 22 days in animals with or without skin grafts; T-cell recovery correlated with skin graft rejection. All three immunotoxins elicited >100 day median survival of fully allogeneic heterotopic heart grafts. By 100 days, T cells recovered to normal numbers in these animals, but the grafts showed chronic rejection. Conclusion. These immunotoxins profoundly deplete T cells in vivo and effectively prolong allogeneic graft survival. C1 Novartis Pharmaceut Corp, Novartis Inst Biomed Res, Summit, NJ 07901 USA. Regeneron Pharmaceut Inc, Tarrytown, NY 10591 USA. Novartis Pharmaceut, Basel, Switzerland. NIMH, NIH, Bethesda, MD 20892 USA. RP Weetall, M (reprint author), Novartis Pharmaceut Corp, Novartis Inst Biomed Res, Summit, NJ 07901 USA. FU PHS HHS [L-113-9910] NR 42 TC 12 Z9 16 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0041-1337 J9 TRANSPLANTATION JI Transplantation PD MAY 27 PY 2002 VL 73 IS 10 BP 1658 EP 1666 AR UNSP 0041-1337/02/7310-1658/0 DI 10.1097/00007890-200205270-00023 PG 9 WC Immunology; Surgery; Transplantation SC Immunology; Surgery; Transplantation GA 560WN UT WOS:000176104500023 PM 12042656 ER PT J AU Ganesh, G AF Ganesh, G TI Regret e-mails may appeal to employers SO CURRENT SCIENCE LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Ganesh, G (reprint author), NIH, Bldg 37,Room 6138, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU CURRENT SCIENCE ASSN PI BANGALORE PA C V RAMAN AVENUE, PO BOX 8005, BANGALORE 560 080, INDIA SN 0011-3891 J9 CURR SCI INDIA JI Curr. Sci. PD MAY 25 PY 2002 VL 82 IS 10 BP 1195 EP 1195 PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 558FW UT WOS:000175956500009 ER PT J AU Baynham, MT Patel, S Moaddel, R Wainer, IW AF Baynham, MT Patel, S Moaddel, R Wainer, IW TI Multidimensional on-line screening for ligands to the alpha 3 beta 4 neuronal nicotinic acetylcholine receptor using an immobilized nicotinic receptor liquid chromatographic stationary phase SO JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES LA English DT Article DE high-throughout screening; immobilized receptors; nicotinic receptor ligands ID KETAMINE AB The alpha3beta4 subtype of the neuronal nicotinic acetylcholine receptor (nAChR) subtype was immobilized on a liquid chromatographic support and the resulting column used for the rapid and direct on-line screening for nAChR ligands. A multidimensional chromatographic system was developed consisting of the immobilized receptor column (NR column) connected via a switching valve to a C-18 column that was, in turn, connected to a single quadrupole mass spectrometer. A mixture of 18 compounds, containing alpha3beta4 nAChR (7) and compounds that are not alpha3beta4 nAChR ligands (11), was injected onto the NR column. The mobile phase consisted of ammonium acetate (10 mM, pH 7.4)-methanol (95:5, v/v) and the flow-rate was 0.2 ml/min. For the first 8 min the eluent was directed to waste. At t = 8 min, the switching valve was rotated and the NR column connected to the C-18 column. The eluent from the NR column was directed to the C-18 column for 12 min. At t = 20 min, the switching valve was rotated and the NR column was disconnected from the C-18 column. The compounds trapped on the C-18 column were separated and eluted onto the mass spectrometer using a mobile phase of ammonium acetate (10 mM, pH 7.4)-methanol (40:60, v/v) at a flow-rate of 1.0 ml/min. Detection was accomplished using total ion monitoring. The multidimensional system correctly isolated six of the seven alpha3beta4 nAChR ligands and only one of the 11 non-ligands was found with the alpha3beta4 nAChR ligands. The results indicate that the multidimensional liquid chromatographic system can be used for the on-line screening of chemical mixtures for alpha3beta4 nAChR ligands. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIA, Bioanalyt & Drug Unit, NIH, Baltimore, MD 21224 USA. RP Wainer, IW (reprint author), NIA, Bioanalyt & Drug Unit, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 9 TC 23 Z9 24 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1570-0232 J9 J CHROMATOGR B JI J. Chromatogr. B PD MAY 25 PY 2002 VL 772 IS 1 BP 155 EP 161 AR PII S1570-0232(02)00070-3 DI 10.1016/S1570-0232(02)00070-3 PG 7 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 558AK UT WOS:000175941700018 PM 12016027 ER PT J AU Graham, BS Rutigliano, JA Johnson, TR AF Graham, BS Rutigliano, JA Johnson, TR TI Respiratory syncytial virus immunobiology and pathogenesis SO VIROLOGY LA English DT Review ID ATTACHMENT GLYCOPROTEIN-G; CELL-MEDIATED-IMMUNITY; T-LYMPHOCYTE ACTIVITY; PULMONARY EOSINOPHILIA; AIRWAY HYPERRESPONSIVENESS; RSV CHALLENGE; G-PROTEIN; IN-VIVO; INFECTION; MICE C1 NIAID, Viral Pathogenesis Lab, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. RP Graham, BS (reprint author), Bldg 40,Room 2502,40 Convent Dr,MSC-3017, Bethesda, MD 20892 USA. NR 51 TC 53 Z9 53 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD MAY 25 PY 2002 VL 297 IS 1 BP 1 EP 7 DI 10.1006/viro.2002.1431 PG 7 WC Virology SC Virology GA 570FT UT WOS:000176648800001 PM 12083830 ER PT J AU Lafrenie, RM Lee, SF Hewlett, IK Yamada, KM Dhawan, S AF Lafrenie, RM Lee, SF Hewlett, IK Yamada, KM Dhawan, S TI Involvement of integrin alpha v beta 3 in the pathogenesis of human immunodeficiency virus type 1 infection in monocytes SO VIROLOGY LA English DT Article DE human immunodeficiency virus type 1; integrin; vitronectin receptor; monocyte/macrophage; pathogenesis ID CULTURED HUMAN MACROPHAGES; CELL-ADHESION; HUMAN MONOCYTES/MACROPHAGES; ENDOTHELIAL-CELLS; HIV-1 INFECTION; AIDS VIRUS; RECEPTOR; EXPRESSION; ENTRY; CD4 AB Attachment of HIV to macrophages is a critical early event in the establishment of infection. In the present study, we demonstrate the involvement of integrin alphavbeta3 (vitronectin receptor) in HIV infection of peripheral blood monocyte-derived macrophages. Culturing monocytes in the presence of M-CSF for 3 days upregulated expression of the alphav-containing integrins, alphavbeta3 and alphavbeta5. The increase in alphavbeta3 expression was accompanied by increased HIV-1 replication by monocytes, Immunoblot analysis showed that purified HIV-gp120 protein interacted with CD4 and alphavbeta3 in immunoprecipitation experiments. Neutralizing antibodies against the alphavbeta3 integrin interfered with the coprecipitation of alphavbeta3 with an anti-gp120 antibody and substantially inhibited HIV infection of monocytes. Neutralizing antibodies against alphavbeta5 or 01 integrins did not significantly affect HIV infection. These results indicate that HIV infection of primary monocytes requires differentiation of these cells and may involve alphavbeta3 interaction with the HIV-1 envelope protein gp120 for productive infection, (C) 2002 Elsevier Science (USA). C1 NE Ontario Reg Canc Ctr, Sudbury, ON P3E 5J1, Canada. US FDA, Ctr Biol Evaluat & Res, Mol Virol Lab, Immunopathogenesis Sect, Rockville, MD 20852 USA. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. RP Lafrenie, RM (reprint author), NE Ontario Reg Canc Ctr, 41 Ramsey Lake Rd, Sudbury, ON P3E 5J1, Canada. NR 49 TC 17 Z9 19 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD MAY 25 PY 2002 VL 297 IS 1 BP 31 EP 38 DI 10.1006/viro.2002.1399 PG 8 WC Virology SC Virology GA 570FT UT WOS:000176648800004 PM 12083833 ER PT J AU Skiadopoulos, MH Surman, SR Riggs, JM Orvell, C Collins, PL Murphy, BR AF Skiadopoulos, MH Surman, SR Riggs, JM Orvell, C Collins, PL Murphy, BR TI Evaluation of the replication and immunogenicity of recombinant human parainfluenza virus type 3 vectors expressing up to three foreign glycoproteins SO VIROLOGY LA English DT Article ID HEMAGGLUTININ-NEURAMINIDASE GLYCOPROTEIN; VESICULAR STOMATITIS VIRUSES; HEXAMER GENOME LENGTH; FUSION GLYCOPROTEINS; MEASLES-VIRUS; ATTENUATION PHENOTYPES; MONOCLONAL-ANTIBODIES; PIV TYPE-1; STABLE EXPRESSION; VACCINE CANDIDATE AB The level of replication and immunogenicity of recombinant parainfluenza virus type 3 (rHPIV3) bearing one, two, or three,gene insertions expressing foreign protective antigens was examined. cDNA-derived recombinant HPIV3s bearing genes encoding the open reading frames (ORFs) of the hemagglutinin-neuraminidase (HN) of HPIV1, the HN of HPIV2, or the hemagglutinin (HA) of measles virus replicated efficiently in vitro, including the largest recombinant, which had three gene unit insertions and which was almost 23 kb in length, 50% longer than unmodified HPIV3. Several viruses were recovered from cDNAs whose genome length was not a multiple of six nucleotides and these contained nucleotide insertions that corrected the length to be a multiple of 6, confirming that the "rule of six" applies to HPIV3, Using a hemagglutination inhibition assay, we determined that the HPIV1 HN expressed by recombinant HPIV3 was incorporated into HPIV3 virions, whereas using this assay incorporation of the HPIV2 HN could not be detected, HPIV3 virions bearing HPIV1 HN were not neutralized by HPIV1 antiserum but were readily neutralized by antibodies to the HPIV3 HN or fusion protein (F). Viruses with inserts were restricted for replication in the respiratory tract of hamsters, and the level of restriction was a function of the total number of genes inserted, the nature of the insert, and the position of the inserted gene in the gene order. A single insert of HPIV2 HN or measles virus HA reduced the in vivo replication of rHPIV3 up to 25-fold, whereas the HPIV1 HN insert decreased replication almost 1000-fold. This indicates that the HPIV1 HN insert has an attenuating effect in addition to that of the extra gene insert itself, presumably because it is incorporated into the virus particle, Viruses containing two inserts were generally more attenuated than those with a single insert, and viruses with three inserts were over-attenuated for replication in hamsters. inserts between the N and P genes were slightly more attenuating than those between the P and the M genes. A recombinant HPIV3 bearing both the HPIV1 and the HPIV2 HN genes (r1 HN 2HN) was attenuated, immunogenic, 3nd protected immunized hamsters from challenge with HPIV1, HPIV2, and HPIV3. Thus, it is possible to use a single HPIV vector expressing two foreign gene inserts to protect infants and young children from the severe lower respiratory tract disease caused by the three major human PIV pathogens, (C) 2002 Elsevier Science (USA). C1 NIAID, Resp Viruses Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. Huddinge Univ Hosp, Dept Clin Virol, S-14186 Stockholm, Sweden. RP Skiadopoulos, MH (reprint author), NIAID, Resp Viruses Sect, Infect Dis Lab, NIH, Bldg 50,Room 6511,50 South Dr,MSC 8007, Bethesda, MD 20892 USA. NR 51 TC 34 Z9 36 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 0042-6822 J9 VIROLOGY JI Virology PD MAY 25 PY 2002 VL 297 IS 1 BP 136 EP 152 DI 10.1006/viro.2002.1415 PG 17 WC Virology SC Virology GA 570FT UT WOS:000176648800015 PM 12083844 ER PT J AU Skiadopoulos, MH Surman, SR Riggs, JM Elkins, WR St Claire, M Nishio, M Garcin, D Kolakofsky, D Collins, PL Murphy, BR AF Skiadopoulos, MH Surman, SR Riggs, JM Elkins, WR St Claire, M Nishio, M Garcin, D Kolakofsky, D Collins, PL Murphy, BR TI Sendai virus, a murine parainfluenza virus type 1, replicates to a level similar to human PIV1 in the upper and lower respiratory tract of African green monkeys and chimpanzees SO VIROLOGY LA English DT Article ID AMINO-ACID-SEQUENCES; HEMAGGLUTININ-NEURAMINIDASE; PARA-INFLUENZA; NIPAH-VIRUS; INFECTION; MICE; PROTEIN; VACCINE; GLYCOPROTEINS; GENOME AB Human parainfluenza virus type 1 (HPIV1), a major cause of croup in infants and young children, accounts for 6% of hospitalizations for pediatric respiratory tract disease. The antigenically related Sendai virus, referred to here as murine PIV1 (MPIV1) is being considered for use as a live-attenuated vaccine to protect against HPIVI1 (J. L. Hurwitz, K. F Soike, M. Y, Sangster, A. Portner, R. E. Sealy, D. H, Dawson, and C Coleclough, 1997, Vaccine 15(5), 533-540) and also as a recombinant vaccine vector expressing antigens to protect against viral disease in humans, However, in the 1960s MPIV1 was reported to have been isolated from humans, suggesting that zoonotic transmission might have occurred. It is therefore important to examine the ability of MPIV1 to replicate in nonhuman primates, i.e., surrogate hosts for humans. In the present study the level of replication of MPIV1 and HPIV1 was compared in African green monkeys and chimpanzees. Surprisingly, vi replicated as efficiently as HPIV1 in the upper and lower respiratory tract of African green monkeys at doses of 10(4) and 10(6) and replicated only slightly less efficiently at both sites in chimpanzees, African green monkeys immunized with MPIV1 were highly resistant to subsequent challenge with HPIV1 even though MPIV1 did not induce a detectable HPIV1-neutralizing antibody response. The high level of replication of MPIV1 observed in the upper and lower respiratory tract of these primates suggests that MPIV1 likely would require significant attenuation before it could be given to humans as a vaccine against HPIV1 or as a vaccine vector. Its ability to efficiently replicate in nonhuman primates suggests that MPIV1 lacks a significant host range restriction in primates and could theoretically cause zoonotic disease in humans. (C) 2002 Elsevier Science (USA). C1 NIAID, Resp Viruses Sect, NIH, Bethesda, MD 20892 USA. NIAID, Expt Primate Virol Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. Bioqual Inc, Rockville, MD 20850 USA. Univ Geneva, Sch Med, CMU, Dept Genet & Microbiol, CH-1211 Geneva, Switzerland. RP Skiadopoulos, MH (reprint author), NIAID, Resp Viruses Sect, NIH, Bldg 50,Room 6511,60 South Dr,MSC 8007, Bethesda, MD 20892 USA. NR 47 TC 37 Z9 38 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD MAY 25 PY 2002 VL 297 IS 1 BP 153 EP 160 DI 10.1006/viro.2002.1416 PG 8 WC Virology SC Virology GA 570FT UT WOS:000176648800016 PM 12083845 ER PT J AU Jacobson, MA Spritzler, J Landay, A Chan, E Katzenstein, D Schock, B Fox, L Roe, J Kundu, S Pollard, R AF Jacobson, MA Spritzler, J Landay, A Chan, E Katzenstein, D Schock, B Fox, L Roe, J Kundu, S Pollard, R CA AACTG 325 Protocol Team TI A phase I, placebo-controlled trial of multi-dose recombinant human interleukin-12 in patients with HIV infection SO AIDS LA English DT Article DE interleukin-12; HIV; AIDS; cytokines; T-helper 1 ID IMMUNODEFICIENCY-VIRUS INFECTION; EXPERIMENTAL VIRAL-INFECTIONS; INTERFERON-GAMMA PRODUCTION; BLOOD MONONUCLEAR-CELLS; IMMUNE-RESPONSES; MYCOBACTERIUM-AVIUM; VITRO; MICE; PROLIFERATION; RESTORATION AB Objectives: Interleukin (IL)-12 is a cytokine that stimulates T lymphocytes and natural killer cells to generate a Type I T-helper lymphocyte immune response. The primary objective of this study was to determine the safety and immunologic activity of repeated recombinant human IL-12 (rhIL-12) dosing in HIV-infected patients over a broad range of the HIV disease spectrum. Design: A randomized, placebo-controlled, Phase I trial design was chosen to control for the effects of HIV disease alone on safety and immunologic measurements. Methods: HIV-infected patients on antiretroviral therapy received rhIL-12 or placebo twice weekly for 4 weeks. Subjects were monitored for safety and changes in absolute lymphocyte subset number, serum interferon (IFN)gamma and neopterin levels, plasma HIV RNA level, peripheral blood mononuclear cell (PBMC)-inducible IFNgamma responses to mitogen, and PBMC proliferative responses to phytohemagglutinin, tetanus, Candida, Mycobacterium avium complex, streptokinase, and HIV p24 and gpl 60 antigens. Results: rhIL-12 was well tolerated at doses up to 100 ng/kg in subjects enrolled with CD4 cell counts < 50 X 10(6) cells/l and at all doses in subjects with CD4 cell counts of 300 x 10(6)-500 X 10(6) cells/l. rhIL-12 resulted in dose-related increases in serum neopterin (particularly in subjects with baseline CD4 cell counts of 300-500 X 106 cells/l) but in no significant changes in other immunologic measurements or plasma HIV RNA levels. Conclusions: rhIL-12 dosed twice weekly at less than or equal to 100 ng/kg was well tolerated in HIV-infected patients and resulted in dose-related increases in serum neopterin (possibly reflecting the effect of some degree of IFNgamma induction). However, there was no evidence of improvement in antigen-specific immune response. (C) 2002 Lippincott Williams Wilkins. C1 Univ Calif San Francisco, Dept Med, Posit Hlth Program, San Francisco, CA USA. San Francisco Gen Hosp, Med Serv, San Francisco, CA 94110 USA. Harvard Univ, Sch Publ Hlth, Boston, MA 02115 USA. Rush Med Coll, Dept Immunol Microbiol, Chicago, IL 60612 USA. Stanford Univ, Dept Med, Stanford, CA 94305 USA. Frontier Sci & Technol Res Fdn Inc, Amherst, NY USA. NIAID, Bethesda, MD 20892 USA. Adult AIDS Clin Trials Grp Operat Ctr, Rockville, MD USA. Univ Texas, Dept Internal Med, Galveston, TX 77555 USA. RP Jacobson, MA (reprint author), Ward 84,Potrero, San Francisco, CA 94110 USA. FU NCRR NIH HHS [RR00044, 5-M01-RR00083]; NIAID NIH HHS [U01 AI25915-10, U01 AI27663, AI-27658]; PHS HHS [UO1 A127659] NR 35 TC 20 Z9 21 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 MAY 24 PY 2002 VL 16 IS 8 BP 1147 EP 1154 DI 10.1097/00002030-200205240-00008 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 554WR UT WOS:000175760600008 PM 12004273 ER PT J AU Kim, JH Nam, KH Kwon, OS Kim, IG Bustin, M Choy, HE Park, SC AF Kim, JH Nam, KH Kwon, OS Kim, IG Bustin, M Choy, HE Park, SC TI Histone cross-linking by transglutaminase SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE transglutaminase; core historic modification; erythrocyte; cross-linking ID TISSUE TRANSGLUTAMINASE; CHROMATIN STRUCTURE; PROTEIN; MODULATION; STAGE; CELLS; MENT AB Transglutaminases irreversibly catalyze covalent cross-linking of proteins by forming isopeptide bonds between peptide-bound glutamine and lysine residues. Among several transglutaminases, tissue-type transglutaminase (tTGase) is most ubiquitously found in every type of cells and tissues in animals, but its natural substrate has yet to be identified. In an attempt to identify the natural substrate for tTGase, we examined in vitro if core histones were subject to cross-linking by tTGase. We found core histone subunits, H2A and H2B, were specifically cross-linked by tTGase. The cross-linking was between either one or both glutalmnes at C-terminal end of H2A (-VTIAQ(104)GGVLPNTQ(112) SVLLPKKTESSKSK-C' end) and the first and/or third lysine from C-terminal end of H2B (-AVESEGK(116)AVTKYTSSK(125)-C' end). The cross-linking occurred only when these subunits were released from nucleosome but not when these were organized in nucleosome. Most interestingly, in chicken erythrocyte the cross-linked H2A-H2B was present in a significant amount. From these results, it can be proposed that tTGase-mediated cross-linking is an another form of core histone modification and it may play a role of chromatin condensation during erythrocyte differentiation. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Seoul Natl Univ, Coll Med, Dept Biochem & Mol Biol, Chongno Gu, Seoul 110799, South Korea. NCI, Lab Mol Carcinogenesis, NIH, Bethesda, MD 20892 USA. Chonnam Natl Univ, Coll Med, Dept Microbiol, Kwangju 501191, South Korea. RP Park, SC (reprint author), Seoul Natl Univ, Coll Med, Dept Biochem & Mol Biol, Chongno Gu, 28 Yongon Dong, Seoul 110799, South Korea. RI Kim, In-Gyu/J-2747-2012; Bustin, Michael/G-6155-2015 NR 30 TC 20 Z9 20 U1 2 U2 3 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 24 PY 2002 VL 293 IS 5 BP 1453 EP 1457 AR PII S0006-291X(02)00393-5 DI 10.1016/S0006-291X(02)00393-5 PG 5 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 560YA UT WOS:000176107900024 PM 12054678 ER PT J AU de Caestecker, MP Bottomley, M Bhattacharyya, S Payne, TL Roberts, AB Yelick, PC AF de Caestecker, MP Bottomley, M Bhattacharyya, S Payne, TL Roberts, AB Yelick, PC TI The novel type I serine-threonine kinase receptor Alk8 binds TGF-beta in the presence of TGF-beta RII SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE activin-like kinase receptor (Alk); serine-threonine kinases; transforming growth factor-beta (TGF-beta); cell signaling pathways ID GROWTH-FACTOR-BETA; FUNCTIONAL-CHARACTERIZATION; ACTIVIN; ZEBRAFISH; ACTIVATION; IDENTIFICATION; SPECIFICITY; SIGNALS AB The TGF-beta superfamily consists of an array of ligands including BMP, TGF-beta, activin, and nodal subfamilies. The extensive range of biological effects elicited by TGF-beta family signaling is due in part to the large numbers and promiscuity of types I and II TGF-beta family member receptors. Alk8 is a novel type I TGF-beta family member receptor first identified in zebrafish [Dev. Dyn. 211 (4) (1998) 352], which participates in BMP signaling pathways [Development 128 (6) (2001) 849; Development 128 (6) (2001) 859 Mech. Dev. 100 (2) (2001) 275; J. Dent. Res. 80 (11) (2001) 1968]. Here we report that Alk8 also forms active signaling complexes with TGF-beta in the presence of TGF-betaRII These results expand the signaling repertoire of zAlk8 by demonstrating an ability to participate in two distinct TGF-beta subfamily signaling pathways. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Forsyth Inst, Dept Oral Biol, Dept Cytokine Biol & Harvard Forsyth, Boston, MA 02115 USA. Vanderbilt Univ, Sch Med, Div Nephrol, Nashville, TN 37232 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Yelick, PC (reprint author), Forsyth Inst, Dept Oral Biol, Dept Cytokine Biol & Harvard Forsyth, 140 The Fenway, Boston, MA 02115 USA. NR 29 TC 7 Z9 7 U1 1 U2 2 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 24 PY 2002 VL 293 IS 5 BP 1556 EP 1565 AR PII S0006-291X(02)00424-2 DI 10.1016/S0006-291X(02)00424-2 PG 10 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 560YA UT WOS:000176107900040 PM 12054694 ER PT J AU Karmakar, P Piotrowski, J Brosh, RM Sommers, JA Miller, SPL Cheng, WH Snowden, CM Ramsden, DA Bohr, VA AF Karmakar, P Piotrowski, J Brosh, RM Sommers, JA Miller, SPL Cheng, WH Snowden, CM Ramsden, DA Bohr, VA TI Werner protein is a target of DNA-dependent protein kinase in vivo and in vitro, and its catalytic activities are regulated by phosphorylation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SYNDROME CELLS; HELICASE ACTIVITY; KU AUTOANTIGEN; WRN PROTEIN; BINDING; COMPLEX; SUBUNIT; REPAIR; MICE; TRANSCRIPTION AB Human Werner Syndrome is characterized by early onset of aging, elevated chromosomal instability, and a high incidence of cancer. Werner protein (WRN) is a member of the recQ gene family, but unlike other members of the recQ family, it contains a unique 3'-->5' exonuclease activity. We have reported previously that human Ku heterodimer interacts physically with WRN and functionally stimulates WRN exonuclease activity. Because Ku and DNA-PKcs, the catalytic subunit of DNA-dependent protein kinase (DNA-PK), form a complex at DNA ends, we have now explored the possibility of functional modulation of WRN exonuclease activity by DNA-PK. We find that although DNA-PKcs alone does not affect the WRN exonuclease activity, the additional presence of Ku mediates a marked inhibition of it. The inhibition of WRN exonuclease by DNA-PKcs requires the kinase activity of DNA-PKcs. WRN is a target for DNA-PKcs phosphorylation, and this phosphorylation requires the presence of Ku. We also find that treatment of recombinant WRN with a Ser/Thr phosphatase enhances WRN exonuclease and helicase activities and that WRN catalytic activity can be inhibited by rephosphorylation of WRN with DNA-PK. Thus, the level of phosphorylation of WRN appears to regulate its catalytic activities. WRN forms a complex, both in vitro and in vivo, with DNA-PKC. WRN is phosphorylated in vivo after treatment of cells with DNA-damaging agents in a pathway that requires DNA-PKcs. Thus, WRN protein is a target for DNA-PK phosphorylation in vitro and in vivo, and this phosphorylation may be a way of regulating its different catalytic activities, possibly in the repair of DNA dsb. C1 NIA, Lab Mol Gerontol, NIH, Baltimore, MD 21224 USA. Univ Calgary, Dept Biochem & Mol Biol, Calgary, AB T2N 1N4, Canada. Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA. Univ N Carolina, Curriculum Genet & Mol Biol, Chapel Hill, NC 27599 USA. RP Bohr, VA (reprint author), NIA, Lab Mol Gerontol, NIH, Box 1,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. FU NCI NIH HHS [CA84442-01, R01 CA084442] NR 52 TC 119 Z9 124 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 MAY 24 PY 2002 VL 277 IS 21 BP 18291 EP 18302 DI 10.1074/jbc.M111523200 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 558PM UT WOS:000175975800008 PM 11889123 ER PT J AU Lavenu-Bombled, C Trainor, CD Makeh, I Romeo, PH Max-Audit, I AF Lavenu-Bombled, C Trainor, CD Makeh, I Romeo, PH Max-Audit, I TI Interleukin-13 gene expression is regulated by GATA-3 in T cells - Role of a critical association of a GATA and two GATG motifs SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TRANSCRIPTION FACTOR GATA-3; TRANSGENIC MICE; DNA-BINDING; EMBRYONIC LETHALITY; ENFORCED EXPRESSION; GAMMA PRODUCTION; NERVOUS-SYSTEM; CUTTING EDGE; TH1 CELLS; ELEMENTS AB Using a transgenic approach, we studied the role of GATA-3 in T cells. As previously shown, enforced GATA-3 expression in transgenic mice inhibits Th1 differentiation of CD4 T cells, but unexpectedly, both type 1 (interferon gamma) and type 2 (interleukin (IL)-4 and IL-13) cytokine genes were activated in the transgenic CD8 T cells. Because IL-13 gene expression was highly enhanced in vivo by GATA-3 expression, we studied the human and the mouse IL-13 gene promoters and found an evolutionary-conserved association of a consensus GATA binding site and two GATG motifs. We showed that efficient GATA-3 binding to this regulatory sequence required these three motifs and that the affinity of the GATA zinc fingers for this association was five times higher than for the consensus GATA binding site alone. Transfections in a T cell line or transactivation by GATA-3 showed that the combination of the three sites was required for full transcriptional activity of the IL-13 gene promoter. Finally we showed that this association of binding sites causes a very high sensitivity of the IL-13 gene promoter to small variations in the level of GATA-3 protein. Altogether, these results indicate an important role of GATA-3 in CD8 cytokine gene expression and demonstrate that a critical network of GATA binding sites highly modulates GATA-3 activity. C1 Univ Paris 05, Matern Port Royal, Dept Hematol, Inst Cochen,CNRS,INSERM, F-75014 Paris, France. NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Max-Audit, I (reprint author), Univ Paris 05, Matern Port Royal, Dept Hematol, Inst Cochen,CNRS,INSERM, 123 Bd Port Royal, F-75014 Paris, France. NR 47 TC 56 Z9 58 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 24 PY 2002 VL 277 IS 21 BP 18313 EP 18321 DI 10.1074/jbc.M110013200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 558PM UT WOS:000175975800010 PM 11893731 ER PT J AU Ray, K Northup, J AF Ray, K Northup, J TI Evidence for distinct cation and calcimimetic compound (NPS 568) recognition domains in the transmembrane regions of the human Ca2+ receptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID METABOTROPIC GLUTAMATE-RECEPTOR; PROTEIN-COUPLED RECEPTORS; CALCIUM-SENSING RECEPTOR; CELL-SURFACE EXPRESSION; EXTRACELLULAR DOMAIN; SIGNAL-TRANSDUCTION; CHIMERIC RECEPTORS; LIGAND-BINDING; IDENTIFICATION; SPECIFICITY AB The Ca2+ receptor, a member of the family 3 of G protein-coupled receptors (GPCR), responds not only to its primary physiological ligand Ca2+ but also to other di- and trivalent metals (Mg2+, Gd3+) and the organic polycations spermine and poly-L-Arginine. As has been found for other family 3 GPCRs, the large amino-terminal extracellular domain (ECD) of the Ca2+ receptor is the primary Ca2+ binding domain. To examine how the signal is propagated from the ECD to the seven-transmembrane core domain (7TM) we constructed a Ca2+ receptor mutant (T903-Rhoc) lacking the entire ECD but containing the 7TM. We have found that this structure initiates signaling in human embryonic kidney (HEK) 293 cells stably expressing the construct. One or more cation recognition sites are also located within the 7TM. Not only Ca2+, but also several other Ca2+ receptor-specific agonists, Mg2+, Gd3+, spermine, and poly-L-Arginine, can activate T903-Rhoc truncated receptor-initiated phosphoinositide hydrolysis in HEK 293 cells. The phenylalkylamine compound, N-PS 568, identified as a positive allosteric modulator of the Ca2+ receptor can selectively potentiate the actions of Ca2+ and other polycationic agonists on the T903-Rhoc receptor. Similarly, organic polycations synergistically activate T903-Rhoc with di- and trivalent metals. Alanine substitution of all the acidic residues in the second extracellular loop of the T903-Rhoc receptor significantly impairs activation by metal ions and organic polycations in the presence of NPS 568 but not the synergistic activation of Ca2+ with poly-L-Arginine. These data indicate that although the ECD has been thought to be the main determinant for Ca2+ recognition, the 7TM core of the Ca2+ receptor contains activating site(s) recognizing Ca2+ and Gd3+ as well as the allosteric modulators NPS 568 and organic polycations that may play important roles in the regulation of receptor activation. C1 NIDCD, Lab Cellular Biol, NIH, Bethesda, MD 20892 USA. RP Ray, K (reprint author), NIDCD, Lab Cellular Biol, 5 Res Ct,Rm 2A-11, Rockville, MD 20850 USA. NR 30 TC 72 Z9 72 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 MAY 24 PY 2002 VL 277 IS 21 BP 18908 EP 18913 DI 10.1074/jbc.M202113200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 558PM UT WOS:000175975800086 PM 11880385 ER PT J AU Gao, ZG Chen, A Barak, D Kim, SK Muller, CE Jacobson, KA AF Gao, ZG Chen, A Barak, D Kim, SK Muller, CE Jacobson, KA TI Identification by site-directed mutagenesis of residues involved in ligand recognition and activation of the human A(3) adenosine receptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROTEIN-COUPLED RECEPTORS; BINDING-SITE; TRANSMEMBRANE DOMAIN; MOLECULAR-CLONING; CEREBRAL-ISCHEMIA; PHOSPHOLIPASE-C; RHODOPSIN; EXPRESSION; NUCLEOSIDES; ANTAGONISTS AB Ligand recognition has been extensively explored in G protein-coupled A(1), A(2A), and A(2B) adenosine receptors but not in the As receptor, which is cerebroprotective and cardioprotective. We mutated several residues of the human A(3) adenosine receptor within transmembrane domains 3 and 6 and the second extracellular loop, which have been predicted by previous molecular modeling to be involved in the ligand recognition, including His(95), Trp(243), Leu(244), Ser(247), Asn(250), and Lys(152). The N250A mutant receptor lost the ability to bind both radiolabeled agonist and antagonist. The H95A mutation significantly reduced affinity of both agonists and antagonists. In contrast, the K152A (EL2), W243A (6.48), and W243F (6.48) mutations did not significantly affect the agonist binding but decreased antagonist affinity by similar to3-38-fold, suggesting that these residues were critical for the high affinity of A(3) adenosine receptor antagonists. Activation of phospholipase C by wild type (WT) and mutant receptors was measured. The As agonist 2-chloro-N-6-(3-iodobenzyl)-5'-N-methylcarbamoyladenosine stimulated phosphoinositide turnover in the WT but failed to evoke a response in cells expressing W243A and W243F mutant receptors, in which agonist binding was less sensitive to guanosine 5'-gamma-thiotriphosphate than in WT. Thus, although not important for agonist binding, Trp(243) was critical for receptor activation. The results were interpreted using a rhodopsin-based model of ligand-A(3) receptor interactions. C1 NIDDK, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Univ Bonn, Inst Pharmaceut, D-53115 Bonn, Germany. RP Jacobson, KA (reprint author), NIDDK, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bldg 8A,Rm B1A-19, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009; Muller, Christa/C-7748-2014 OI Jacobson, Kenneth/0000-0001-8104-1493; Muller, Christa/0000-0002-0013-6624 NR 49 TC 120 Z9 121 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 MAY 24 PY 2002 VL 277 IS 21 BP 19056 EP 19063 DI 10.1074/jbc.M110960200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 558PM UT WOS:000175975800106 PM 11891221 ER PT J AU Lapidus, LJ Eaton, WA Hofrichter, J AF Lapidus, LJ Eaton, WA Hofrichter, J TI Measuring dynamic flexibility of the coil state of a helix-forming peptide SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE alpha-helix; alanine peptide; polyalanine; molecular dynamics; contact rate ID ALANINE-BASED PEPTIDES; UNFOLDED CYTOCHROME-C; ALPHA-HELIX; FAST EVENTS; CONTACT FORMATION; ENERGY-TRANSFER; FOLDING PROBLEM; KINETICS; DIFFUSION; MECHANISMS AB To investigate the dynamic flexibility of the coil state of a helix-forming peptide the end-to-end contact rate was determined. Nanosecond optical excitation of tryptophan at one end of a 22 residue, alanine peptide populates a long-lived triplet state which is quenched upon close contact with a cyclic disulfide attached to the opposite end. Analysis of the decay of the triplet population using a two-state model for helix formation yields the diffusion-limited end-to-end contact rate of the coil state of the peptide as well as the helix - coil and coil - helix rates. The helix-coil rates are very similar to those previously measured in laser temperature-jump experiments. The end-to-end contact rate of 1.1 X 10(7) s(-1) in the coil state is tenfold faster than the rate for a disordered peptide with threonine substituted for alanine and, somewhat surprisingly, is about twice the rate for a disordered glycine-containing peptide. These differences are discussed in terms of the theory of Szabo, Schulten and Schulten. The rates should provide important new benchmarks for testing the accuracy of atomistic molecular dynamics simulations. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 NIDDKD, Dept Chem Phys, NIH, Bethesda, MD 20892 USA. RP Eaton, WA (reprint author), NIDDKD, Dept Chem Phys, NIH, Bldg 5,5 Ctr Dr, Bethesda, MD 20892 USA. NR 48 TC 40 Z9 40 U1 2 U2 8 PU ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD MAY 24 PY 2002 VL 319 IS 1 BP 19 EP 25 DI 10.1016/S0022-2836(02)00193-6 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 559UA UT WOS:000176042700003 PM 12051933 ER PT J AU Ghribi, O Herman, MM Savory, J AF Ghribi, O Herman, MM Savory, J TI The endoplasmic reticulum is the main site for caspase-3 activation following aluminum-induced neurotoxicity in rabbit hippocampus SO NEUROSCIENCE LETTERS LA English DT Article DE apoptosis; caspase-3; endoplasmic reticulum; cytosol; calnexin; aluminum; hippocampus ID CYTOCHROME-C; CELL-DEATH; APOPTOSIS; BCL-2; BAX; NEURODEGENERATION; TRANSLOCATION; MITOCHONDRIA; PROTEASE; DISEASE AB We have assessed the distribution of caspase-3 in subcellular fractions from rabbit brain hippocampus and find that in controls the pro-caspase-3 form is distributed mainly in the cytoplasm. In animals treated intracisternally with the neurotoxin aluminum-maltolate, although pro-caspase-3 levels are higher in the cytosolic fractions, p17, the active caspase-3, is localized mainly in the endoplasmic reticulum. This distribution is confirmed by immunohistochemistry which demonstrates the co-localization of p17 with calnexin, a specific marker of the endoplasmic reticulum. Based on the apparent translocation into the endoplasmic reticulum of active caspase-3, an executioner of cell death, the results suggest that this organelle is an important site in the caspase-3 mediated apoptosis cascade. Inhibition of the latter enzyme by directly targeting its main site of activation could represent a strategy to prevent this adverse event. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Virginia, Hlth Sci Ctr, Dept Pathol, Charlottesville, VA 22908 USA. Univ Virginia, Dept Biochem & Mol Genet, Charlottesville, VA 22908 USA. NIMH, IRP, NIH, Bethesda, MD 20892 USA. RP Savory, J (reprint author), Univ Virginia, Hlth Sci Ctr, Dept Pathol, Box 168, Charlottesville, VA 22908 USA. NR 20 TC 30 Z9 32 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 MAY 24 PY 2002 VL 324 IS 3 BP 217 EP 221 AR PII S0304-3940(02)00147-7 DI 10.1016/S0304-3940(02)00147-7 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 566ZN UT WOS:000176458300013 PM 12009527 ER PT J AU Sommer, MA Wurtz, RH AF Sommer, MA Wurtz, RH TI A pathway in primate brain for internal monitoring of movements SO SCIENCE LA English DT Article ID FRONTAL EYE FIELD; COROLLARY DISCHARGE; SUPERIOR COLLICULUS; EFFERENCE COPY; POSITION; RESPONSES; SACCADES; MUSCLES; NEURONS; SYSTEM AB It is essential to keep track of the movements we make, and one way to do that is to monitor correlates, or corollary discharges, of neuronal movement commands. We hypothesized that a previously identified pathway from brainstem to frontal cortex might carry corollary discharge signals. We found that neuronal activity in this pathway encodes upcoming eye movements and that inactivating the pathway impairs sequential eye movements consistent with loss of corollary discharge without affecting single eye movements. These results identify a pathway in the brain of the primate Macaca mulatta that conveys corollary discharge signals. C1 NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. RP Sommer, MA (reprint author), NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. NR 29 TC 311 Z9 313 U1 1 U2 10 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 MAY 24 PY 2002 VL 296 IS 5572 BP 1480 EP 1482 DI 10.1126/science.1069590 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 557LP UT WOS:000175910300052 PM 12029137 ER PT J AU Le Stunff, H Peterson, C Liu, H Milstien, S Spiegel, S AF Le Stunff, H Peterson, C Liu, H Milstien, S Spiegel, S TI Sphingosine-l-phosphate and lipid phosphohydrolases SO BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS LA English DT Review DE sphingolipid; sphingosine-1-phosphate; sphingosine; ceramide; lipid phosphate phosphohydrolase; S1P phosphohydrolase; sphingosine kinase ID LONG-CHAIN BASES; MIGRATING GERM-CELLS; SACCHAROMYCES-CEREVISIAE; MOLECULAR-CLONING; SPHINGOLIPID METABOLISM; SPHINGOSINE-1-PHOSPHATE LYASE; FUNCTIONAL-CHARACTERIZATION; SIGNAL-TRANSDUCTION; MAMMALIAN-CELLS; PHOSPHOLIPASE-D AB Sphingosine-1-phosphate (SIP) is a bioactive sphingolipid that acts as both an extracellular ligand for the endothelial differentiation gene-1 (EDG-1) G-protein coupled receptor (GPCR) family and as an intracellular messenger. Cellular levels of S1P are low and tightly regulated in a spatial-temporal manner not only by sphingosine kinase (SPHK) but also by degradation catalyzed by SIP lyase, specific SIP phosphohydrolases, and by general lipid phosphate phosphohydrolases (LPPs). LPPs are characterized as magnesium-independent, insensitive to inhibition by N-ethylmaleimide (NEW and possessing broad substrate specificity with a variety of phosphorylated lipids, including SIP, phosphatidic acid (PA), and lysophosphatidic acid (LPA). LPPs contain three highly conserved domains that define a phosphohydrolase superfamily. Recently, several specific SIP phosphohydrolases have been identified in yeast and mammalian cells. Phylogenetic and biochemical analyses indicate that these enzymes constitute a new subset of the LPP family. As further evidence, SIP phosphohydrolases exhibit high specificity for phosphorylated sphingoid bases. Enforced expression of SIP phosphohydrolase alters the cellular levels of sphingolipid metabolites in yeast and mammalian cells, increasing sphingosine and ceramide, bioactive sphingolipids that often have opposing biological actions to SIP. By regulating the cellular ratio between ceramide/sphingosine and SIP, SIP phosphohydrolase is poised to be a critical factor in cell survival/cell death decisions. Indeed, expression of SIP phosphohydrolase in mammalian cells increases apoptosis, whereas deletion of SIP phosphohydrolases in yeast correlates with resistance to heat stress. In this review, we discuss the role of phosphohydrolases in the metabolism of SIP and how turnover of SIP can regulate sphingolipid metabolites signaling. (C) 2002 Elsevier Science B.V All rights reserved. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Biochem, Richmond, VA 23298 USA. Georgetown Univ, Med Ctr, Dept Biochem & Mol Biol, Washington, DC 20007 USA. NIMH, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. RP Spiegel, S (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Biochem, 1101 E Marshall St, Richmond, VA 23298 USA. FU NIGMS NIH HHS [GM43880] NR 65 TC 65 Z9 66 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1388-1981 J9 BBA-MOL CELL BIOL L JI Biochim. Biophys. Acta Mol. Cell Biol. Lipids PD MAY 23 PY 2002 VL 1582 IS 1-3 SI SI BP 8 EP 17 AR PII S1388-1981(02)00132-4 DI 10.1016/S1388-1981(02)00132-4 PG 10 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 568BZ UT WOS:000176523700003 PM 12069805 ER PT J AU Allende, ML Proia, RL AF Allende, ML Proia, RL TI Sphingosine-1-phosphate receptors and the development of the vascular system SO BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS LA English DT Review DE angiogenesis; sphingosine-l-phosphate; G protein-coupled receptor; sphingolipid; vascular smooth muscle; endothelial cell ID SPHINGOSINE 1-PHOSPHATE RECEPTOR; PROTEIN-COUPLED RECEPTOR; BLOOD-VESSEL FORMATION; SMOOTH-MUSCLE CELLS; MULTIPLE SIGNALING PATHWAYS; INDUCE ANGIOGENESIS INVITRO; HUMAN ENDOTHELIAL-CELLS; LYSOPHOSPHATIDIC ACID; SPHINGOLIPID METABOLISM; TYROSINE KINASE AB Extracellular sphingolipid signaling has been implicated as an essential event in vascular development. Sphingosine-1-phosphate (S1P), through interactions with G protein-coupled receptors, regulates functions of endothelial and smooth muscle cells (SMCs)-the major cell types of the vasculature. The knockout of the gene encoding the S1P(1) receptor (formally known as Edg-1) in mice blocks vascular maturation, the process where SMCs and pericytes envelop nascent endothelial tubes. The question. that remains is how stimulation of S1P receptors controls this critical event in the developmental sequence leading to the formation of functional blood vessels. Published by Elsevier Science B.V. C1 NIDDK, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. RP Proia, RL (reprint author), NIDDK, Genet Dev & Dis Branch, NIH, Room 9N-314,Bldg 10,10 Ctr DR MSC 1821, Bethesda, MD 20892 USA. RI Proia, Richard/A-7908-2012 NR 85 TC 98 Z9 100 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1388-1981 J9 BBA-MOL CELL BIOL L JI Biochim. Biophys. Acta Mol. Cell Biol. Lipids PD MAY 23 PY 2002 VL 1582 IS 1-3 SI SI BP 222 EP 227 AR PII S1388-1981(02)00175-0 DI 10.1016/S1388-1981(02)00175-0 PG 6 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 568BZ UT WOS:000176523700030 PM 12069832 ER PT J AU Kim, SA Marshall, MA Melman, N Kim, HS Muller, CE Linden, J Jacobson, KA AF Kim, SA Marshall, MA Melman, N Kim, HS Muller, CE Linden, J Jacobson, KA TI Structure-activity relationships at human and rat A(2B) adenosine receptors of xanthine derivatives substituted at the 1-, 3-, 7-, and 8-positions SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID A2-ADENOSINE RECEPTORS; A1-ADENOSINE RECEPTORS; SELECTIVE ANTAGONISTS; RADIOLIGAND BINDING; HIGH-AFFINITY; MAST-CELLS; CAFFEINE; ANALOGS; POTENT; THEOPHYLLINE AB In the search for improved selective antagonist ligands of the A(2B) adenosine receptor, which have the potential as antiasthmatic or antidiabetic drugs, we have synthesized and screened a variety of alkylxanthine derivatives substituted at the 1-, 3-, 7-, and 8-positions. Competition for I-125-ABOPX(I-125-3-(4-amino-3-iodobenzyl)-8-(phenyl-4-oxyacetate)-1-propylxanthine) binding in membranes of stably transfected HEK-293 cells revealed uniformly higher affinity (<10-fold) of these xanthines for human than for rat A(2B) adenosine receptors. Binding to rat brain membranes expressing A(1) and A(2A) adenosine receptors revealed greater A(2B) selectivity over A(2A) than A(1) receptors. Substitution at the 1-position with 2-phenylethyl (or alkyl/olefinic groups) and at N-3 with hydrogen or methyl favored A(2B) selectivity. Relative. to enprofylline 2b, pentoxifylline 35 was equipotent and 1-propylxanthine 3 was >13-fold more potent at human A(2B) receptors. Most N-7 substituents did not enhance affinity over hydrogen, except for 7-(2-chloroethyl), which enhanced the affinity of theophylline by 6.5-fold to 800 nM. The A2B receptor affinity-enhancing effects of 7-(2-chloroethyl) vs 7-methyl were comparable to the known enhancement produced by an 8-aryl substitution. Among 8-phenyl analogues, a larger alkyl group at the 1-position than at the 3-position favored affinity at the human A2B receptor, as indicated by 1-allyl-3-methyl-8-phenylxanthine, with a K-i value of 37 nM. Substitution on the 8-phenyl ring indicated that an electron-rich ring was preferred for A2B receptor binding. In conclusion, new leads for the design of xanthines substituted in the 1-, 3-, 7-, and 8-positions as A(2B) receptor-selective antagonists have been identified. C1 NIDDK, Mol Recognit Sect, Lab Bioorgan Chem, NIH, Bethesda, MD 20892 USA. Univ Virginia, Dept Internal Med, Hlth Sci Ctr, Charlottesville, VA 22908 USA. Univ Virginia, Dept Mol Physiol & Biol Phys, Hlth Sci Ctr, Charlottesville, VA 22908 USA. Univ Bonn, Pharmazeut Inst Pharmazeut Chem, D-5300 Bonn, Germany. RP Jacobson, KA (reprint author), NIDDK, Mol Recognit Sect, Lab Bioorgan Chem, NIH, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009; Muller, Christa/C-7748-2014 OI Jacobson, Kenneth/0000-0001-8104-1493; Muller, Christa/0000-0002-0013-6624 NR 37 TC 62 Z9 64 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD MAY 23 PY 2002 VL 45 IS 11 BP 2131 EP 2138 DI 10.1021/jm0104318 PG 8 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 554AY UT WOS:000175711700003 PM 12014951 ER PT J AU Yordanov, AT Yamada, K Krishna, MC Russo, A Yoo, J English, S Mitchell, JB Brechbiel, MW AF Yordanov, AT Yamada, K Krishna, MC Russo, A Yoo, J English, S Mitchell, JB Brechbiel, MW TI Acyl-protected hydroxylamines as spin label generators for EPR brain imaging SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID ELECTRON-PARAMAGNETIC-RESONANCE; FREE-RADICALS; NITROXIDE RADICALS; POLYNITROXYL-ALBUMIN; EPILEPTIC SEIZURES; OXIDATIVE STRESS; MURINE TUMOR; RAT-HEART; VIVO; METABOLISM AB In a search for novel electron paramagnetic resonance (EPR) brain imaging agents, we have designed and synthesized the acyl-protected hydroxylamines 1-acetoxy-4-methoxycarbonyl-2,2,6,6-tetramethylpiperidine (AMCPe), 1-acetoxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (AMCPy), and 1-acetoxy-3-(acetoxymethoxy)carbonyl-2,2,5,5-tetramethylpyrrolidine (DACPy), in which both the ring size and the number of ester functions were varied. In all of them, the nitroxide was first reduced and the resultant hydroxylamine was then protected with an acetyl group. These compounds are lipophilic, which is a major prerequisite for blood-brain barrier penetration. Once in the brain, esterases and oxidants quickly convert these derivatives into ionic, water-soluble radicals and thus EPR detectable species that then reside in the central nervous system for periods of time sufficient for detection and imaging. The biological relevancy of these new compounds in mice has been assessed, and their biodistribution patterns have been compared. The five-membered ring derivative AMCPy emerged as a potent EPR brain imaging agent while the other two derivatives, AMCPe and DACPy, were quite ineffective. C1 NCI, Radioimmune & Inorgan Chem Sect, NIH, Bethesda, MD 20892 USA. NCI, Radiat Biol Branch, Canc Res Ctr, NIH, Bethesda, MD 20892 USA. RP Yordanov, AT (reprint author), NCI, Radioimmune & Inorgan Chem Sect, NIH, Bethesda, MD 20892 USA. RI Yamada, Ken-ichi/E-6318-2012 NR 34 TC 12 Z9 12 U1 0 U2 0 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 MAY 23 PY 2002 VL 45 IS 11 BP 2283 EP 2288 DI 10.1021/jm0105169 PG 6 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 554AY UT WOS:000175711700018 PM 12014966 ER PT J AU Stubbs, MC Hall, DJ AF Stubbs, MC Hall, DJ TI The amino-terminus of the E2F-1 transcription factor inhibits DNA replication of autonomously replicating plasmids in mammalian cells SO ONCOGENE LA English DT Article DE S phase checkpoint; E2F-1; replication inhibition; elongation phase ID VIRUS-40 CORE ORIGIN; ZINC-FINGER PROTEIN; S-PHASE CHECKPOINT; DAMAGE RESPONSE; APOPTOSIS; INDUCTION; EXPRESSION; INITIATION; BINDING; BRCA1 AB The E2F1 transcription factor plays a pivotal role in driving cells out of a quiescent state and into the S phase of the cell cycle, in part by transactivating genes needed for DNA replication including DHFR, thymidine kinase, and DNA Polymerase x. E2F1 has also been implicated in regulating an S phase checkpoint, however its role in this checkpoint is not well defined. To determine how E2F1 affects such a checkpoint, we utilized an in vivo replication assay employing a plasmid based SV40 origin of replication, transfected into cells expressing SV40 large T antigen. Here we show that expression of full length E2F1, or only its N terminus, represses replication from plasmids containing the SV40 origin, while N terminal deletions of E2F1 do not. E2F1 appears to inhibit the elongation phase of replication and not the initiation phase since it does not affect the replication of other cotransfected plasmids containing only the SV40 origin. Further, inhibition of replication is dependent on both the amino-terminus of the E2F1 protein and on a DNA sequence that is contained within the 3' end of the E2F1 cDNA. Additionally, both full-length E2F1, or just its N-terminus, form protein complexes with two portions of the 3' end of the E2F1 cDNA. These data provide a clue to the mechanism by which E2F1 regulates transit through the S phase checkpoint, by acting on a specific DNA sequence via its amino-terminal region, to inhibit elongation of DNA replication. C1 NIAMSD, Cartilage & Orthopaed Branch, NIH, Bethesda, MD 20892 USA. RP Hall, DJ (reprint author), NIAMSD, Cartilage & Orthopaed Branch, NIH, MSC 5755,9000 Rockville Pk, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA67032] NR 44 TC 3 Z9 3 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 23 PY 2002 VL 21 IS 23 BP 3715 EP 3726 DI 10.1038/sj/onc/1205473 PG 12 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 553LE UT WOS:000175676000007 PM 12032840 ER PT J AU List, K Haudenschild, CC Szabo, R Chen, WJ Wahl, SM Swaim, W Engelholm, LH Behrendt, N Bugge, TH AF List, K Haudenschild, CC Szabo, R Chen, WJ Wahl, SM Swaim, W Engelholm, LH Behrendt, N Bugge, TH TI Matriptase/MT-SP1 is required for postnatal survival, epidermal barrier function, hair follicle development, and thymic homeostasis SO ONCOGENE LA English DT Article DE apoptosis; cell-surface proteolysis; skin; thymus; barrier function ID HEPATOCYTE GROWTH-FACTOR; TRANSMEMBRANE SERINE-PROTEASE; FACTOR HGF ACTIVATOR; T-CELL SELECTION; PLASMINOGEN-ACTIVATOR; KERATINOCYTE TRANSGLUTAMINASE; LAMELLAR ICHTHYOSIS; LINKED ICHTHYOSIS; STRATUM-CORNEUM; IN-VITRO AB Matriptase/MT-SP1 is a novel tumor-associated type II transmembrane serine protease that is highly expressed in the epidermis, thymic stroma, and other epithelia. A null mutation was introduced into the Matriptase/MT-SP1 gene of mice to determine the role of Matriptase/MT-SP1 in epidermal development and neoplasia. Matriptase/MT-SP1-deficient mice developed to term but uniformly died within 48 h of birth. All epidermal surfaces of newborn mice were grossly abnormal with a dry, red, shiny, and wrinkled appearance. Matriptase/MT-SP1-deficiency caused striking malformations of the stratum corneum, characterized by dysmorphic and pleomorphic corneocytes and the absence of vesicular bodies in transitional layer cells. This aberrant skin development seriously compromised both inward and outward epidermal barrier function, leading to the rapid and fatal dehydration of Matriptase/MT-SP1-deficient pups. Loss of Matriptase/MT-SP1 also seriously affected hair follicle development resulting in generalized follicular hypoplasia, absence of erupted vibrissae, lack of vibrissal hair canal formation, ingrown vibrissae, and wholesale abortion of vibrissal follicles. Furthermore, Matriptase/MT-SP1-deficiency resulted in dramatically increased thymocyte apoptosis, and depletion of thymocytes. This study demonstrates that Matriptase/MT-SP1 has pleiotropic functions in the development of the epidermis, hair follicles, and cellular immune system. C1 Natl Inst Dent & Craniofacial Res, Proteases & Tissue Remodeling Unit, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Cellular Imaging Core, NIH, Bethesda, MD 20892 USA. Finsen Lab, DK-2100 Copenhagen, Denmark. Amer Red Cross, Dept Expt Pathol, Rockville, MD 20855 USA. RP Bugge, TH (reprint author), Natl Inst Dent & Craniofacial Res, Proteases & Tissue Remodeling Unit, Oral & Pharyngeal Canc Branch, NIH, 30 Convent Dr,Room 211, Bethesda, MD 20892 USA. NR 60 TC 215 Z9 218 U1 2 U2 11 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 23 PY 2002 VL 21 IS 23 BP 3765 EP 3779 DI 10.1038/sj/onc/1205502 PG 15 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 553LE UT WOS:000175676000011 PM 12032844 ER PT J AU Nelson, DE Bland, S Powell-Griner, E Klein, R Wells, HE Hogelin, G Marks, JS AF Nelson, DE Bland, S Powell-Griner, E Klein, R Wells, HE Hogelin, G Marks, JS TI State trends in health risk factors and receipt of clinical preventive services among US adults during the 1990s SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID UNITED-STATES; SELF-REPORT; BELT USE; MAMMOGRAPHY; PREVALENCE; OBESITY; CANCER; MORTALITY; POPULATION; OVERWEIGHT AB Context Monitoring trends is essential for evaluating past activities and guiding current preventive health program and policy efforts. Although tracking progress toward national health goals is helpful, use of national estimates is limited because most preventive health care activities, policies, and other efforts occur at the state or community level. There may be important state trends that are obscured by national data. Objective To estimate state-specific trends for 5 health risk factors and 6 clinical preventive services. Design Telephone surveys were conducted from 1991 through 2000 as part of the Behavioral Risk Factor Surveillance System. Setting and Participants Randomly selected adults aged 18 years or older from 49 US states. Annual state sample sizes ranged from 1188 to 7543. Main Outcome Measures Statistically significant changes (P<.01) instate prevalences of cigarette smoking, binge alcohol use, physical inactivity, obesity, safety belt use, and mammography; screening for cervical cancer, colorectal cancer, and cholesterol levels; and receipt of influenza and pneumococcal disease vaccination. Results There were statistically significant increases in safety belt use for 39 of 47 states and receipt of mammography in the past 2 years for women aged 40 years or older for 43 of 47 states. For persons aged 65 years or older, there were increases in receipt of influenza vaccination for 44 of 49 states and ever receiving pneumococcal vaccination for 48 of 49 states. State trends were mixed for binge alcohol use (increasing in 19 of 47 states and declining in 3), physical inactivity (increasing in 3 of 48 states and declining in 11), and cholesterol screening (increasing in 13 of 47 states and decreasing in 5). Obesity increased in all states and smoking increased in 14 of 47 states (declining only in Minnesota). Cervical cancer screening increased in 8 of 48 states and colorectal cancer screening increased in 13 of 49 states. New York experienced improvements for 8 of 11 measures, while 7 of 11 measures improved in Delaware, Kentucky, and Maryland; in contrast, Alaska experienced improvements for no measures and at least 4 of 11 measures worsened in Iowa, North Dakota, and South Dakota. Conclusions Most states experienced increases in safety belt use, mammography, and adult vaccinations. Trends for smoking and binge alcohol use are disturbing, and obesity data support previous findings. Trend data are useful for targeting state preventive health efforts. C1 Ctr Dis Control & Prevent, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA USA. Natl Ctr Hlth Stat, Hyattsville, MD 20782 USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. RP Nelson, DE (reprint author), NCI, Div Canc Control & Populat Sci, 6130 Execut Blvd,MSC 7365,EPN 4068, Bethesda, MD 20892 USA. EM nelsond@mail.nih.gov NR 70 TC 101 Z9 106 U1 4 U2 6 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60654-0946 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD MAY 22 PY 2002 VL 287 IS 20 BP 2659 EP 2667 DI 10.1001/jama.287.20.2659 PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 554JQ UT WOS:000175732400025 PM 12020301 ER PT J AU Domanski, M Mitchell, G Pfeffer, M Neaton, JD Norman, J Svendsen, K Grimm, R Cohen, J Stamler, J AF Domanski, M Mitchell, G Pfeffer, M Neaton, JD Norman, J Svendsen, K Grimm, R Cohen, J Stamler, J CA MRFIT Res Grp TI Pulse pressure and cardiovascular disease-related mortality - Follow-up study of the Multiple Risk Factor Intervention Trial (MRFIT) SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID ISOLATED SYSTOLIC HYPERTENSION; CORONARY HEART-DISEASE; MIDDLE-AGED MEN; BLOOD-PRESSURE; MYOCARDIAL-INFARCTION; POPULATION; PREDICTOR; WOMEN; DEATH AB Context The sixth Joint National Committee (JNC-VI) classification system of blood pressure emphasizes both systolic blood pressure (SBP) and diastolic blood pressure (DBP) for cardiovascular disease risk assessment. Pulse pressure may also be a valuable risk assessment tool. Objective To compare relationships of SBP, DBP, and pulse pressure, separately and jointly, with cardiovascular disease-related mortality in men. Design and Setting Data from the Multiple Risk Factor Intervention Trial (MRFIT), which screened men aged 35 to 57 years from 1973 through 1975 at 22 US centers, was used to assess cardiovascular disease-related mortality through 1996. Participants A total of 342815 men without diabetes or a history of myocardial infarction were divided into 2 groups based on their age at MRFIT screening (35- to 44-year-olds and 45- to 57-year olds). Participant blood pressure levels were classified into a JNC-VI blood pressure category based on SBP and DBP (optimal, normal but not optimal, high normal, stage 1 hypertension, stage 2-3 hypertension), and pulse pressure was calculated. Main Outcome Measure Cardiovascular disease-related mortality. Results There were 25721 cardiovascular disease-related deaths. Levels of SBP and DBP were more strongly related to cardiovascular disease than pulse pressure. Relationships of SBP, DBP, and pulse pressure to cardiovascular disease-related mortality varied within JNC-VI category. Concordant elevations of SBP and DBP were associated with a greater risk of cardiovascular disease-related mortality for both age groups of men, Among men aged 45 to 57 years, higher SBP and lower DBP (discordant elevations) also yielded a greater risk of cardiovascular disease-related mortality. Conclusion In both age groups, cardiovascular disease risk assessment was improved by considering both SBP and DBP, not just SBP, DBP, or pulse pressure separately. C1 Univ Minnesota, Sch Publ Hlth, Div Biostat, Minneapolis, MN 55414 USA. NHLBI, Clin Trials Grp, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, Bethesda, MD 20892 USA. Cardiovasc Engn Inc, Holliston, MA USA. Brigham & Womens Hosp, Div Cardiol, Boston, MA 02115 USA. Univ Minnesota, Hennepin Cty Med Ctr, Berman Ctr Outcomes & Clin Res, Minneapolis, MN 55415 USA. St Louis Univ, Sch Med, Dept Internal Med, St Louis, MO USA. Northwestern Univ, Sch Med, Dept Prevent Med, Chicago, IL USA. RP Neaton, JD (reprint author), Univ Minnesota, Sch Publ Hlth, Div Biostat, 2221 Univ Ave SE,Room 200, Minneapolis, MN 55414 USA. FU NHLBI NIH HHS [R01-HL68140-02] NR 34 TC 149 Z9 154 U1 0 U2 5 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 MAY 22 PY 2002 VL 287 IS 20 BP 2677 EP 2683 DI 10.1001/jama.287.20.2677 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 554JQ UT WOS:000175732400027 PM 12020303 ER PT J AU Otsuka, C Kobayashi, K Kawaguchi, N Kunitomi, N Moriyama, K Hata, Y Iwai, S Loakes, D Noskov, VN Pavlov, Y Negishi, K AF Otsuka, C Kobayashi, K Kawaguchi, N Kunitomi, N Moriyama, K Hata, Y Iwai, S Loakes, D Noskov, VN Pavlov, Y Negishi, K TI Use of yeast transformation by oligonucleotides to study DNA lesion bypass in vivo SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE oligonucleotide transformation; Saccharomyces cerevisiae; nucleoside analog; (6-4)photoproduct; REV1 ID ESCHERICHIA-COLI; SYNTHETIC OLIGONUCLEOTIDES; SACCHAROMYCES-CEREVISIAE; MAMMALIAN-CELLS; POLYMERASE-ETA; PHOTOPRODUCT; MUTAGENESIS; PYRIMIDINE; OLIGODEOXYNUCLEOTIDES; MECHANISMS AB We have studied mutagenic specificities of DNA lesions in vivo in yeast CYC1 oligonuclecitide transformation assay. We introduced two lesions into oligonucleotides. One was a nucleoside analog, 3,4-dihydro-6H,8H-pyrimido[4,5-c][1,2]oxazin-7-one 2'-deoxyriboside (dP), which is highly mutagenic to bacteria. It is supposed to be a miscoding, but otherwise good template for DNA polymerases. The other lesion was the TT pyrimidine(6-4)pyrimidone photoproduct, one of the typical UV lesions, which blocks DNA replication. These oligonucleotides were used to transform yeast cycl mutants with ochre nonsense mutation to Cyc1(+). As expected from its templating proper-Lies in vitro, the transforming activity of dP-containing oligonucleotides was similar to those of unmodified oligonucteotides. Results indicated that dP may direct incorporation of guanine and adenine at a ratio of 1:20 or more in vivo. An oligonucleotide containing the photoproduct showed the transforming activity of as low as 3-5% of that of the corresponding unmodified oligonucleotide. This bypass absolutely required REV1 gene. The sequence analysis of the transformants has shown that the lesion was read as TT and TC at a ratio of 3:7, indicating its high mutagenic potential. (C) 2002 Elsevier Science B.V. All rights reserved. C1 Okayama Univ, Ctr Gene Res, Okayama 7008530, Japan. Biomol Engn Res Inst, Suita, Osaka 5650874, Japan. MRC, Mol Biol Lab, Cambridge CB2 2QH, England. NCI, Bethesda, MD 20892 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Negishi, K (reprint author), Okayama Univ, Ctr Gene Res, Okayama 7008530, Japan. NR 25 TC 17 Z9 17 U1 1 U2 3 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 MAY 22 PY 2002 VL 502 IS 1-2 BP 53 EP 60 AR PII S0027-5107(02)00023-4 DI 10.1016/S0027-5107(02)00023-4 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 557YN UT WOS:000175936800007 PM 11996972 ER PT J AU Lodmell, DL Parnell, MJ Bailey, JR Ewalt, LC Hanlon, CA AF Lodmell, DL Parnell, MJ Bailey, JR Ewalt, LC Hanlon, CA TI Rabies DNA vaccination of non-human primates: post-exposure studies using gene gun methodology that accelerates induction of neutralizing antibody and enhances neutralizing antibody titers SO VACCINE LA English DT Article DE rabies virus; DNA vaccine; booster; neutralizing antibody; post-exposure protection ID PROTECTIVE IMMUNITY; VIRUS-INFECTION; EAR PINNA; MICE; IMMUNIZATION; PROPHYLAXIS; CELLS; DOGS; SUPERIORITY; CHLOROQUINE AB Pre-exposure DNA vaccination protects non-human primates against rabies virus. Post-exposure protection of monkeys against rabies virus by DNA vaccination has not been attempted. Presumably, post-exposure experiments have not been undertaken because neutralizing antibody is usually slow to be induced after DNA vaccination. In this study, we initially attempted to accelerate the induction of neutralizing antibody by varying the route and site of DNA vaccination and booster frequency, Gene gun (GG) vaccinations above axillary and inguinal lymph nodes or in ear pinnae generated higher levels of neutralizing antibody than intradermal (ID) needle vaccinations in the pinnae. Concurrent GG booster vaccinations above axillary and inguinal lymph nodes and in ear pinnae, 3 days after primary vaccination, accelerated detectable neutralizing antibody. GG booster vaccinations also resulted in higher neutralizing antibody levels and increased the durability of this response. Post-exposure vaccination with DNA or the human diploid cell vaccine (HDCV), in combination with an one-time treatment with human rabies immune 'globulin (HRIG), protected 50 and 75% of the monkeys, respectively, as compared to 75% mortality of the controls. These data will be useful for the refinement, development, and implementation of future pre- and post-exposure rabies DNA vaccination studies. Published by Elsevier Science Ltd. C1 NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, Hamilton, MT 59840 USA. Ctr Dis Control & Prevent, Natl Ctr Infect Dis, Div Viral & Rickettsial Dis, Viral & Rickettsial Zoonoses Branch,Rabies Sect, Atlanta, GA 30333 USA. RP Lodmell, DL (reprint author), NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, 903 S 4th St, Hamilton, MT 59840 USA. NR 44 TC 34 Z9 34 U1 0 U2 5 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 MAY 22 PY 2002 VL 20 IS 17-18 BP 2221 EP 2228 AR PII S0264-410X902)00143-3 DI 10.1016/S0264-410X(02)00143-3 PG 8 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 560QK UT WOS:000176092600010 PM 12009276 ER PT J AU Hou, YC Hu, WG Hirano, T Gu, XX AF Hou, YC Hu, WG Hirano, T Gu, XX TI A new intra-NALT route elicits mucosal and systemic immunity against Moraxella catarrhalis in a mouse challenge model SO VACCINE LA English DT Article DE intra-NALT; mucosal immunization; Moraxella catarrhalis ID OUTER-MEMBRANE PROTEIN; BRANHAMELLA-CATARRHALIS; LYMPHOID-TISSUE; STREPTOCOCCUS-PNEUMONIAE; PULMONARY CLEARANCE; IMMUNOGLOBULIN-A; OTITIS-MEDIA; IMMUNIZATION; RESPONSES; ANTIBODIES AB Mucosally administered antigens are often poorly immunogenic due to the difficulty of transporting antigens through the mucosal epithelium. We investigated a new route of intranasal-associated lymphoid tissue (intra-NALT) administration of antigens to circumvent the antigen transportation barrier. A comparative study was carried out on mice administered with killed whole cells of Moraxella catarrhalis strain-25238 plus cholera toxin (CT) by intra-NALT injection and nasal inoculation. Both routes induced significant elevations of several isotype antibodies against strain 25238 in saliva, lung lavage, and serum as measured by an enzyme-linked immunosorbent assay (ELISA). Most of these antibodies were paralleled by the numbers of their corresponding antibody forming cells in mucosal or systemic lymphoid tissues. However, intra-NALT injection elicited higher levels of immunoglobulin (Ig) A and IgG in saliva, IgA and IgG in lung lavage, and IgG and IgM in sera than nasal inoculation (P less than or equal to 0.05). In addition, both routes generated significant reductions of bacteria in lungs following an aerosol challenge with strain 25238 in a mouse model of pulmonary clearance. Once again, intra-NALT route showed better bacterial clearance in mouse lungs than nasal inoculation (P < 0.01). These results demonstrate that intra-NALT administration of antigens is a convenient and effective route for mucosal immunization that elicits improved mucosal and systemic immunity. This new route can be used as a model to study mucosal antigens or vaccine candidates for antigen activation and interaction with the NALT that is one of major inductive sites for common mucosal immune system. Published by Elsevier Science Ltd. C1 Natl Inst Deafness & Other Commun Disorders, NIH, Rockville, MD 20850 USA. RP Gu, XX (reprint author), Natl Inst Deafness & Other Commun Disorders, NIH, 5 Res Court, Rockville, MD 20850 USA. NR 36 TC 26 Z9 27 U1 0 U2 0 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 MAY 22 PY 2002 VL 20 IS 17-18 BP 2375 EP 2381 AR PII S0264-410X(02)00097-X DI 10.1016/S0264-410X(02)00097-X PG 7 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 560QK UT WOS:000176092600028 PM 12009294 ER PT J AU Lawson, VA Oelrichs, R Guillon, C Imrie, AA Cooper, DA Deacon, NJ McPhee, DA AF Lawson, VA Oelrichs, R Guillon, C Imrie, AA Cooper, DA Deacon, NJ McPhee, DA TI Adaptive changes after human immunodeficiency virus type 1 transmission SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID ANTIBODY-DEPENDENT ENHANCEMENT; BLOOD MONONUCLEAR-CELLS; EVOLUTION IN-VIVO; HIV-1 INFECTION; MONOCLONAL-ANTIBODIES; BIOLOGICAL PHENOTYPE; POSITIVE INDIVIDUALS; VIRAL PHENOTYPE; AIDS PATIENTS; HIGH TITERS AB Primary HIV-1 infection (PHI) is associated with a period of viremia, the resolution of which generally coincides with the development of both humoral and cellular immune responses. In this study replication-competent quasispecies were derived from virus isolated from an individual before and after seroconversion. Virus was also isolated from the presumed donor. Phenotypic and genotypic analysis of biological clones identified transmission of an R5/M-tropic phenotype. However, the ability of clones derived from the recipient to replicate in primary macrophages and PBMCs was restricted after transmission. This apparent selection process was supported by analysis of molecular clones derived from the isolated virus. Analysis of the ratio of synonymous and nonsynonymous substitutions predicted the existence of selective pressure soon after transmission, coincident with the development of HIV-1-specific antibodies. An Env trans-complementation assay demonstrated that the infectivity of a clone derived from the recipient after seroconversion was enhanced in the presence of a selected neutralizing antibody, indicating that the developing humoral immune response may have at least in part contributed to the selective pressure identified. C1 Macfarlane Burnet Inst Med Res & Publ Hlth, AIDS Cellular Biol Unit, Fairfield, Vic 3078, Australia. Macfarlane Burnet Inst Med Res & Publ Hlth, Natl Ctr HIV Virol Res, Fairfield, Vic 3078, Australia. Univ Melbourne, Dept Microbiol & Immunol, Parkville, Vic 3052, Australia. Macfarlane Burnet Inst Med Res & Publ Hlth, AIDS Mol Biol Unit, Fairfield, Vic 3078, Australia. Thai Red Cross Soc Program AIDS, Bangkok, Thailand. Erasmus Univ, Dept Virol, NL-3000 DR Rotterdam, Netherlands. Dept Immunol, Sydney, NSW, Australia. Natl Ctr HIV Epidemiol & Clin Res, Sydney, NSW, Australia. RP Lawson, VA (reprint author), NIAID, Persistent Viral Dis Lab, Rocky Mt Labs, NIH, 903 S 4th St, Hamilton, MT 59840 USA. EM vlawson@niad.nih.gov RI guillon, christophe/A-4084-2015; OI guillon, christophe/0000-0001-6378-3303; Lawson, Victoria/0000-0002-7362-7176 NR 64 TC 5 Z9 5 U1 0 U2 1 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD MAY 20 PY 2002 VL 18 IS 8 BP 545 EP 556 DI 10.1089/088922202753747897 PG 12 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 554JG UT WOS:000175731600003 PM 12036484 ER PT J AU Krasnoselskaya-Riz, I Spruill, A Chen, YW Schuster, D Teslovich, T Baker, C Kumar, A Stephan, DA AF Krasnoselskaya-Riz, I Spruill, A Chen, YW Schuster, D Teslovich, T Baker, C Kumar, A Stephan, DA TI Nuclear factor 90 mediates activation of the cellular antiviral expression cascade SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID DOUBLE-STRANDED-RNA; INTERFERON REGULATORY FACTOR-3; ENHANCER-BINDING FACTOR-3; TRANSCRIPTIONAL INDUCTION; OLIGONUCLEOTIDE ARRAYS; CELLS; GENE; ALPHA; SYNTHETASE; RESPONSES AB Viral infection triggers a cascade of interferon response genes, but the mechanisms that prime such innate antiviral defenses are poorly understood. Among candidate cellular mediators of the antiviral response are the double-stranded RNA (dsRNA)-binding proteins. Here we show that a C-terminal variant of the ubiquitous dsRNA-binding protein, nuclear factor 90 (NF90ctv), can activate the interferon response genes in the absence of viral infection. NF90ctv-expressing cells were infected with the syncytium-inducing HIV-1 strain NL4-3 and were shown to inhibit viral replication. To gain insight into this mechanism of protection, we analyzed the expression profiles of NF90ctv-positive cells as compared with parental cells transduced with the empty vector. Of the 5600 genes represented on the expression arrays, 90 displayed significant (4-fold or more) changes in mRNA levels in NF90-expressing cells. About 50% are known interferon alpha/beta-stimulated genes. The microarray expression data were confirmed by quantitative reverse transcriptase-polymerase chain reaction analysis of six representative interferon-inducible genes. Electrophoretic mobility shift assays showed that the biological response is mediated by the activation of transcription factors in NF90ctv-expressing cells. Functional significance of the activated transcription complex was evaluated by transfection assays with luciferase reporter constructs driven by the interferon-inducible promoter from the 2'-5'-oligoadenylate synthetase (p69) gene. Resistance to HIV-1, caused by the expression of NF90ctv in the cell culture system, appears to be mediated in part by the induction of interferon response genes. This leads to a hypothesis as to the mechanism of action of NF90 in mediating endogenous antiviral responses. C1 George Washington Univ, Sch Med & Hlth Sci, Dept Biochem & Mol Biol, Washington, DC 20037 USA. Howard Univ, Coll Med, Dept Microbiol, Washington, DC 20059 USA. Childrens Natl Med Ctr, Med Genet Res Ctr, Washington, DC 20010 USA. George Washington Univ, Sch Med, Genet Program, Washington, DC 20037 USA. Quanta Biosci Inc, Rockville, MD 20850 USA. NCI, Cellular Regulat & Transformat Sect, Basic Res Lab, NIH, Bethesda, MD 20892 USA. RP Kumar, A (reprint author), George Washington Univ, Sch Med & Hlth Sci, Dept Biochem & Mol Biol, Ross Hall,2300 Eye St NW, Washington, DC 20037 USA. FU NHLBI NIH HHS [U01 HL66614-01] NR 43 TC 33 Z9 33 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD MAY 20 PY 2002 VL 18 IS 8 BP 591 EP 604 DI 10.1089/088922202753747941 PG 14 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 554JG UT WOS:000175731600008 PM 12036489 ER PT J AU Kitazono, M Chuman, Y Aikou, T Fojo, T AF Kitazono, M Chuman, Y Aikou, T Fojo, T TI Adenovirus HSV-TK construct with thyroid-specific promoter: Enhancement of activity and specificity with histone deacetylase inhibitors and agents modulating the cAMP pathway SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE thyroid cancer; thyroglobulin; adenoviries; suicide gene therapy; histone deacetylase inhibitor ID CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE PATHWAY; DOXORUBICIN PLUS CISPLATIN; MEDIATED GENE-THERAPY; WILD-TYPE P53; SUICIDE GENE; IN-VIVO; ANAPLASTIC CARCINOMA; CANCER CELLS; HEPATOCELLULAR-CARCINOMA; THYROGLOBULIN PROMOTER AB The successful use of tissue- or tumor-selective promoters in targeted gene therapy for cancer depends on high and selective activity. Tg is a thyroid-specific protein that is expressed in the normal thyroid and a majority of thyroid tumors. In the present study, we show, using a luciferase reporter assay, that a construct containing the putative Tg promoter and enhancer is active in 4 thyroid carcinoma cell lines (including 2 anaplastic thyroid carcinoma cell lines) and not in 5 cancer cell lines arising from nonthyroid tissues. Furthermore, both the activity and the specificity of this construct were increased by pretreatment with 8-Br-cAMP and the histone deacetylase inhibitor depsipeptide (FR901228). Expression of thymidine kinase in thyroid cancer cells infected with a recombinant adenovirus (Ad) carrying a Tg enhancer/promoter-thymidine kinase expression cassette (AdTg enhancer/promoter-TK) correlated with the level of Tg enhancer/promoter activity in these cells. Under similar conditions, TK expression was not observed in cancer cell lines arising from nonthyroid tissues. Cells infected with AdTg enhancer/promoter-TK demonstrated preferential GCV sensitivity, with up to a 100,000-fold increase in GCV sensitivity in thyroid cancer cell lines compared to cancer cell lines of nonthyroid origin. The construct described herein can be used to selectively target thyroid cancer cells, and its expression can be modulated to further increase its specificity and selectivity, especially in anaplastic thyroid carcinoma cells, using B-Br-cAMP and depsipeptide. Published 2002 Wiley-Liss, Inc. C1 NCI, NIH, Med Branch, Bethesda, MD 20892 USA. Kagoshima Univ, Fac Med, Dept Surg 1, Kagoshima 890, Japan. RP Fojo, T (reprint author), NCI, NIH, Med Branch, Bldg 10,Room 12N226,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Ain, Kenneth/A-5179-2012 OI Ain, Kenneth/0000-0002-2668-934X NR 49 TC 16 Z9 16 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD MAY 20 PY 2002 VL 99 IS 3 BP 453 EP 459 DI 10.1002/ijc.10307 PG 7 WC Oncology SC Oncology GA 548TB UT WOS:000175404900020 PM 11992417 ER PT J AU Fernandez, T Amoroso, S Sharpe, S Jones, GM Bliskovski, V Kovalchuk, A Wakefield, LM Kim, SJ Potter, M Letterio, JJ AF Fernandez, T Amoroso, S Sharpe, S Jones, GM Bliskovski, V Kovalchuk, A Wakefield, LM Kim, SJ Potter, M Letterio, JJ TI Disruption of transforming growth factor beta signaling by a novel ligand-dependent mechanism SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE receptor; trafficking; intracellular; signal-transduction; plasmacytoma ID BREAST-CANCER CELLS; RECEPTOR-TYPE-II; TGF-BETA; CARCINOMA-CELLS; DOWN-REGULATION; HOST-DEFENSE; EXPRESSION; MICE; GENE; ACTIVATION AB Transforming growth factor (TGF)-beta is the prototype in a family of secreted proteins that act in autocrine and paracrine pathways to regulate cell development and function. Normal cells typically coexpress TGF-beta receptors and one or more isoforms of TGF-beta, thus the synthesis and secretion of TGF-beta as an inactive latent complex is considered an essential step in regulating the activity of this pathway. To determine whether intracellular activation of TGF-beta results in TGF-beta ligand-receptor interactions within the cell, we studied pristane-induced plasma cell tumors (PCTs). We now demonstrate that active TGF-beta1 in the PCT binds to intracellular TGF-beta type II receptor (TbetaRII). Disruption of the expression of TGF-beta1 by antisense TGF-beta1 mRNA restores localization of TbetaRII at the PCT cell surface, indicating a ligand-induced impediment in receptor trafficking. We also show that retroviral expression of a truncated, dominant-negative TbetaRII (dnTbetaRII) effectively competes for intracellular binding of active ligand in the PCT and restores cell surface expression of the endogenous TbetaRII Analysis of TGF-beta receptor-activated Smad2 suggests the intracellular ligand-receptor complex is not capable of signaling. These data are the first to demonstrate the formation of an intracellular TGF-beta-receptor complex, and define a novel mechanism for modulating the TGF-beta signaling pathway. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. RP Letterio, JJ (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bldg 41,Room C629,41 Lib Dr, Bethesda, MD 20892 USA. NR 50 TC 28 Z9 32 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 MAY 20 PY 2002 VL 195 IS 10 BP 1247 EP 1255 DI 10.1084/jem.20011521 PG 9 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 560ZH UT WOS:000176110900002 PM 12021305 ER PT J AU Xu, AM Choi, KL Wang, Y Parmana, PA Xu, LY Bogardus, C Cooper, GJS AF Xu, AM Choi, KL Wang, Y Parmana, PA Xu, LY Bogardus, C Cooper, GJS TI Identification of novel putative membrane proteins selectively expressed during adipose conversion of 3T3-L1 cells SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE adipogenesis; proteomics; insulin; metabolism; membrane protein ID GLUCOSE-TRANSPORTER GLUT4; INSULIN-RESISTANCE; PPAR-GAMMA; ADIPOCYTE; OBESITY; ADIPOGENESIS; PATHWAYS; ADIPSIN; SURFACE AB Fat tissue plays a critical role in the regulation of energy metabolism. Here we report the proteomic identification of a novel fat tissue-specific low molecular weight protein (Falp) which responds to insulin. Falp is preferentially expressed in adipocytes but not in preadipocytes, as shown by two-dimensional gel electrophoresis. Northern blot analysis shows that the Falp gene is predominantly expressed in brown and white fat tissues, but not in any other tissues examined. Human homologs of mouse Falp are found to exist as two alternatively spliced isoforms, which share the same N-terminus but have different C-termini. Both human and mouse Falp contain a conserved putative transmembrane domain. Immunofluorescent analyses of 3T3-L1 adipocytes show that Falp protein strictly localizes at a compact perinuclear membrane compartment. Treatment of cells with insulin induces the redistribution of Falp into numerous discrete spotty structures spreading throughout the cytoplasm. Whereas the function of Falp is currently unclear, its tissue specific expression and the responsiveness to insulin suggest that Falp might be involved in a process specifically restricted to adipose tissue function, such as vesicular transport and protein secretion. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Univ Auckland, Sch Biol Sci, Auckland 1, New Zealand. NIDDKD, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ 85016 USA. RP Xu, AM (reprint author), Univ Auckland, Sch Biol Sci, Private Bag 92019, Auckland 1, New Zealand. RI Xu, Aimin/D-3291-2013; Cooper, Garth /I-3201-2015; OI Xu, Aimin/0000-0002-0668-033X; Cooper, Garth /0000-0001-5241-3163; wang, yu/0000-0001-8697-2940 NR 32 TC 32 Z9 35 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 17 PY 2002 VL 293 IS 4 BP 1161 EP 1167 AR PII S0006-291X(02)00354-6 DI 10.1016/S0006-291X(02)00354-6 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 557MG UT WOS:000175911900003 PM 12054497 ER PT J AU Sun, JX Sui, XX Bradbury, JA Zeldin, DC Conte, MS Liao, JK AF Sun, JX Sui, XX Bradbury, JA Zeldin, DC Conte, MS Liao, JK TI Inhibition of vascular smooth muscle cell migration by cytochrome P450 epoxygenase-derived eicosanoids SO CIRCULATION RESEARCH LA English DT Article DE cytochrome P450; epoxyeicosatrienoic acids; cAMP; protein kinase A; smooth muscle cells ID DEPENDENT PROTEIN-KINASE; EPOXYEICOSATRIENOIC ACIDS; CORONARY-ARTERIES; ARACHIDONIC-ACID; 11,12-EPOXYEICOSATRIENOIC ACID; HYPERPOLARIZING FACTORS; ADP-RIBOSYLATION; ENDOTHELIUM; SYNTHASE; CAMP AB Vascular smooth muscle cell (SMC) migration and proliferation contribute to neointimal hyperplasia and restenosis after vascular injury. The epoxyeicosatrienoic acids (EETs). which are products of cytochrome P450 (CYP) epoxygenases, possess vasodilatory, antiinflammatory, and fibrinolytic properties. To determine whether these compounds also possess antimigratory and antiproliferative properties, we stimulated rat aortic SMCs with either 20% serum or platelet-derived growth factor (PDGF-BB, 20 ng/mL). In a concentration-dependent manner, treatment with EETs, particularly 11,12-EET, inhibited SMC migration through a modified transwell filter by 53% to 60%. EETS, however, have no inhibitory effects on PDGF-stimulated SMC proliferation. Adenoviral-niediated overexpression of the CYP isoform, CYP2J2. in SMCs also inhibited serum- and PDGF-induced SMC migration by 32% and 26%. respectively; both effects of which were reversed by the CYP inhibitors SKF525A or clotrimazole, but not by the K-Ca channel blocker, charybdotoxin. or the cyclooxygenase inhibitor. diclofenac. The effect of EETs correlated with increases in intracellular cAMP levels. Indeed, forskolin and 8-bromo-cAMP exert similar inhibitory effects on SMC migration as 11,12-EET and the effects of 11,12-EET were blocked by cAMP and protein kinase A (PKA) inhibitors. These findings indicate that EETs possess antimigratory effects on SMCs through the cAMP-PKA pathway and suggest that CYP epoxygenase-derived eicosanoids may play important roles in vascular disease and remodeling. C1 Brigham & Womens Hosp, Surg Unit, Cambridge, MA 02139 USA. Harvard Univ, Sch Med, Boston, MA USA. NIEHS, Div Intramural Res, NIH, Res Triangle Pk, NC USA. RP Liao, JK (reprint author), Brigham & Womens Hosp, Surg Unit, 65 Landsdowne St,Room 275, Cambridge, MA 02139 USA. FU NHLBI NIH HHS [HL-52233, HL-04189, HL-48743, K08 HL004189] NR 42 TC 122 Z9 131 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7330 J9 CIRC RES JI Circ.Res. PD MAY 17 PY 2002 VL 90 IS 9 BP 1020 EP 1027 DI 10.1161/01.RES.0000017727.35930.33 PG 8 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 554XF UT WOS:000175761900013 PM 12016269 ER PT J AU Gao, ZG Jacobson, KA AF Gao, ZG Jacobson, KA TI 2-chloro-N-6-cyclopentyladeno sine, adenosine A(1) receptor agonist, antagonizes the adenosine A(3) receptor SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE adenosine derivative; purine; cAMP; phospholipase C; guanine nucleotide ID SITE-DIRECTED MUTAGENESIS; RAT-BRAIN; DERIVATIVES; MODULATION; A(2A); ACTIVATION; BINDING; LIGANDS AB The potent adenosine A(1) receptor agonists, N-6-cyclopentyladenosine (CPA) and 2-chloro-N-6-cyclopentyladenosine (CCPA), were studied in Chinese hamster ovary (CHO) cells expressing the human adenosine A(3) receptor. CPA, but not CCPA, induced phosphoinositide turnover. CPA inhibited forskolin-stimulated cyclic AMP production (EC50 value of 242+/-47 nM). CCPA competitively antagonized the effects of agonist Cl-IB-MECA (2-chloro-N-6-(3-iodobenzyl)-5'-N-methylcarbamoyladenosine) with K-B value of 5.0 nM. CPA competition curves versus the A(3) antagonist radioligand [H-3]PSB-11 (8-ethyl-4-methyl-2-phenyl-(8R)-4,5,7,8-tetrahydro-1H-imidazo[2.1-i]purin-5-one) were right-shifted four-fold by 100 muM GTP, which had no effect on binding of CCPA or the antagonist MRS 1220 (N-[9-chloro-2-(2-furanyl)[1,2,4]triazolo[1,5-c]quinazolin-5-yl]benzene-acetamide). Thus, CCPA is a moderately potent antagonist (K-1=38 nM) of the human A(3) adenosine receptor. Published by Elsevier Science B.V. C1 NIDDKD, Bioorgan Chem Lab, Mol Recognit Sect, NIH, Bethesda, MD 20892 USA. RP Jacobson, KA (reprint author), NIDDKD, Bioorgan Chem Lab, Mol Recognit Sect, NIH, Bldg 8A,Room B1A-19, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 26 TC 17 Z9 17 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD MAY 17 PY 2002 VL 443 IS 1-3 BP 39 EP 42 AR PII S0014-2999(02)01552-2 DI 10.1016/S0014-2999(02)01552-2 PG 4 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 564BE UT WOS:000176293500006 PM 12044789 ER PT J AU Crawford, KW Coop, A Bowen, WD AF Crawford, KW Coop, A Bowen, WD TI sigma(2) receptors regulate changes in sphingolipid levels in breast tumor cells SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE sigma receptor; ceramide; apoptosis ID LIGANDS AB sigma(2) Receptors induce apoptosis in various cell types. The sphingolipid, ceramide as well as the sphingoid bases are involved in cell proliferation. Sphingolipids of MCF-7/Adr- and T47D breast tumor cells were metabolically radiolabeled. The sigma(2) receptor agonists (+)-1R,5R-E-8-(3,4-dichlorobenzylidene)-5-(3-hydroxyphenyl)-2-methylmorphan-7-one (CB-184) and 1S,2R-(-)-cis-N-[2-(3,4-dichlorophenyl) ethyl]-N-methyl-2-(1-pyrrolidinyl)-cyclohexylamine (BD737) caused dose-dependent increases in [H-3]ceramide, with concomitant decreases in [H-3]sphingomyelin. Both effects were attenuated by the novel sigma(2) receptor antagonist, N-phenethylpiperidine oxalate (AC927). sigma(2) Receptors may produce effects on cell growth and apoptosis by regulating the sphingolipid pathway. (C) 2002 Elsevier Science B.V. All rights reserved. C1 NIDDK, Unit Receptor Biochem & Pharmacol, Med Chem Lab, NIH, Bethesda, MD 20892 USA. Howard Univ, Coll Med, Dept Pharmacol, Washington, DC 20059 USA. Univ Maryland, Sch Pharm, Dept Pharmaceut Sci, Baltimore, MD 21201 USA. RP Bowen, WD (reprint author), NIDDK, Unit Receptor Biochem & Pharmacol, Med Chem Lab, NIH, Bldg 8,Room B1-23,8 Ctr Dr MSC 0815, Bethesda, MD 20892 USA. NR 8 TC 61 Z9 64 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD MAY 17 PY 2002 VL 443 IS 1-3 BP 207 EP 209 AR PII S0014-2999(02)01581-9 DI 10.1016/S0014-2999(02)01581-9 PG 3 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 564BE UT WOS:000176293500029 PM 12044812 ER PT J AU Wei, WL Wang, XT Kusiak, JW AF Wei, WL Wang, XT Kusiak, JW TI Signaling events in amyloid beta-peptide-induced neuronal death and insulin-like growth factor I protection SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ACTIVATED PROTEIN-KINASE; N-TERMINAL KINASE; MATURE HIPPOCAMPAL-NEURONS; C-JUN; ALZHEIMERS-DISEASE; PRECURSOR PROTEIN; INDUCED APOPTOSIS; CORTICAL-NEURONS; KAPPA-B; PHOSPHATIDYLINOSITOL 3-KINASE AB Amyloid beta-peptide (Abeta) is implicated as the toxic agent in Alzheimer's disease and is the major component of brain amyloid plaques. In vitro, Abeta causes cell death, but the molecular mechanisms are unclear. We analyzed the early signaling mechanisms involved in Abeta toxicity using the SH-SY5Y neuroblastoma cell line. Abeta caused cell death and induced a 2- to 3-fold activation of JNK JNK activation and cell death were inhibited by overexpression of a dominant-negative SEK1 (SEK1-AL) construct. Butyrolactone I, a cdk5 inhibitor, had an additional protective effect against Abeta toxicity in these SEK1-AL-expressing cells suggesting that cdk5 and JNK activation independently contributed to this toxicity. Abeta also weakly activated ERK and Akt but had no effect on p38 kinase. Inhibitors of ERK and phosphoinositide 3-kinase (PI3K) pathways did not affect Abeta-induced cell death, suggesting that these pathways were not important in Abeta toxicity. Insulin-like growth factor I protected against Abeta toxicity by strongly activating ERK and Akt and blocking JNK activation in a PI3K-dependent manner. Pertussis toxin also blocked Abeta-induced cell death and JNK activation suggesting that G(i/o) proteins were upstream activators of JNK. The results suggest that activation of the JNK pathway and cdk5 may be initial signaling cascades in Abeta-induced cell death. C1 NIA, Mol Neurobiol Unit, Cellular & Mol Biol Lab, Intramural Res Program,Gerontol Res Ctr,NIH, Baltimore, MD 21224 USA. RP Wei, WL (reprint author), NIA, Mol Neurobiol Unit, Cellular & Mol Biol Lab, Intramural Res Program,Gerontol Res Ctr,NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 69 TC 145 Z9 154 U1 0 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 17 PY 2002 VL 277 IS 20 BP 17649 EP 17656 DI 10.1074/jbc.M111704200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100034 PM 11882652 ER PT J AU Cui, Y Miyoshi, K Claudio, E Siebenlist, UK Gonzalez, FJ Flaws, J Wagner, KU Hennighausen, L AF Cui, Y Miyoshi, K Claudio, E Siebenlist, UK Gonzalez, FJ Flaws, J Wagner, KU Hennighausen, L TI Loss of the peroxisome proliferation-activated receptor gamma (PPAR gamma) does not affect mammary development and propensity for tumor formation but leads to reduced fertility SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN BREAST-CANCER; IN-VITRO; TERMINAL DIFFERENTIATION; INDUCE APOPTOSIS; RETINOIC ACID; LUTEIN CELLS; EXPRESSION; LIGAND; MICE; GENE AB The peroxisome proliferation-activated receptor gamma (PPARgamma) is expressed in many cell types including mammary epithelium, ovary, macrophages, and Band T-cells. PPARgamma has an anti-proliferative effect in pre-adipocytes and mammary epithelial cells, and treatment with its ligands reduced the progression of carcinogen-induced mammary tumors in mice. Because PPARgamma-null mice die in utero it has not been possible to study its role in development and tumorigenesis in vivo. To investigate whether PPARgamma is required for the establishment and physiology of different cell types, a cell-specific deletion of the gene was carried out in mice using the Cre-loxP recombination system. We deleted the PPARgamma gene in mammary epithelium using WAP-Cre transgenic mice and in epithelial cells, B- and T-cells, and ovary cells using MMTV-Cre mice. The presence of PPARgamma was not required for functional development of the mammary gland during pregnancy and for the establishment of B- and T-cells. In addition, no increase in mammary tumors was observed. However, loss of the PPARgamma gene in oocytes and granulosa cells resulted in impaired fertility. These mice have normal populations of follicles, they ovulate and develop corpora lutea. Although progesterone levels are decreased and implantation rates are reduced, the exact cause of the impaired fertility remains to be determined. C1 NIDDK, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, Immune Activat Sect, NIH, Bethesda, MD 20892 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Dept Epidemiol & Prevent Med, Baltimore, MD 21201 USA. Univ Nebraska, Med Ctr, Eppley Inst Res Canc & Allied Dis, Omaha, NE 68198 USA. Univ Tokushima, Sch Dent, Dept Biochem, Tokushima 7708504, Japan. RP Hennighausen, L (reprint author), NIDDK, Lab Genet & Physiol, NIH, Bldg 8,Rm 107,8 Ctr Dr, Bethesda, MD 20892 USA. RI Wagner, Kay-Uwe/B-6044-2009 NR 43 TC 117 Z9 123 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 MAY 17 PY 2002 VL 277 IS 20 BP 17830 EP 17835 DI 10.1074/jbc.M200186200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100057 PM 11884400 ER PT J AU Seo, YW Sanyal, S Kim, HJ Won, DH An, JY Amano, T Zavacki, AM Kwon, HB Shi, YB Kim, WS Kang, H Moore, DD Choi, HS AF Seo, YW Sanyal, S Kim, HJ Won, DH An, JY Amano, T Zavacki, AM Kwon, HB Shi, YB Kim, WS Kang, H Moore, DD Choi, HS TI FOR, a novel orphan nuclear receptor related to farnesoid X receptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HORMONE-RECEPTOR; RETINOIC ACID; BILE-ACIDS; GENE; IDENTIFICATION; BINDING; SUPERFAMILY; TRANSCRIPTION; MEMBER; SPECIFICITY AB We have identified and characterized a new amphibian orphan member of the nuclear receptor superfamily and termed it FOR1 (farnesoid X receptor (EXR)-like Orphan Receptor) because it shares the highest amino acid identity with the mammalian FXR. We also identified a variant of FOR1, called FOR2, which has 15 additional C-terminal amino acids. Both variants include an unusual insertion of 33 amino acids in the helix 7 region of the canonical ligand binding domain sequence, suggesting a unique structure for FOR. Northern blot analysis demonstrates that the FOR gene is highly expressed in adult and tadpole liver, kidney, and tail bud stage of the embryo. Detailed expression analysis using in situ hybridization indicates that FOR expression is first detectable at stage 30/31 in the presumptive liver region lasting until stage 41 with a peak level evident at stage 35/36. FOR forms heterodimeric complexes with retinoid X receptor (RXR) as demonstrated by biochemical and mammalian two-hybrid approaches. Gel mobility shift assays demonstrate that FORs form specific DNA-protein complexes on an FXR binding element consisting of an inverted repeat DNA element with 1 nucleotide spacing (IR1) from the phospholipid transfer protein gene promoter. Finally, although FORs do not exhibit constitutive transcriptional activity, frog gallbladder extract significantly augments the transcriptional activities of FORs. C1 Chonnam Natl Univ, Hormone Res Ctr, Kwangju 500757, South Korea. Chonnam Natl Univ, Dept Biol, Kwangju 500757, South Korea. Sogang Univ, Dept Life Sci, Seoul 121742, South Korea. Seoul Natl Univ, Sch Earth & Environm Sci, Marine Biotechnol Lab, Seoul 151, South Korea. NICHHD, Mol Embryol Lab, NIH, Bethesda, MD 20892 USA. Harvard Univ, Brigham & Womens Hosp, Sch Med, Div Thyroid, Boston, MA 02115 USA. Baylor Coll Med, Dept Cellular & Mol Biol, Houston, TX 77030 USA. RP Choi, HS (reprint author), Chonnam Natl Univ, Hormone Res Ctr, Kwangju 500757, South Korea. RI Sanyal, Sabyasachi/E-8034-2010 OI Sanyal, Sabyasachi/0000-0002-8603-3828 NR 35 TC 25 Z9 25 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 17 PY 2002 VL 277 IS 20 BP 17836 EP 17844 DI 10.1074/jbc.M111795200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100058 PM 11867625 ER PT J AU Mukherjee, S Gurevich, VV Preninger, A Hamm, HE Bader, MF Fazleabas, AT Birnbaumer, L Hunzicker-Dunn, M AF Mukherjee, S Gurevich, VV Preninger, A Hamm, HE Bader, MF Fazleabas, AT Birnbaumer, L Hunzicker-Dunn, M TI Aspartic acid 564 in the third cytoplasmic loop of the luteinizing hormone/choriogonadotropin receptor is crucial for phosphorylation-independent interaction with arrestin2 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID OVARIAN FOLLICULAR MEMBRANES; 3RD INTRACELLULAR LOOP; CHORIONIC-GONADOTROPIN RECEPTOR; PROTEIN-COUPLED RECEPTORS; LUTROPIN-CHORIOGONADOTROPIN RECEPTOR; AGONIST-INDUCED PHOSPHORYLATION; BETA-ADRENERGIC-RECEPTOR; RIBOSYLATION FACTOR ARF; ADP-RIBOSYLATION; LUTROPIN/CHORIOGONADOTROPIN RECEPTOR AB Arrestin2 binding to the active but unphosphorylated luteinizing hormone/choriogonadotropin receptor (LH/CG R) in ovarian follicles is triggered by activation of ADP-ribosylation factor 6 (ARF6) and leads to uncoupling of this receptor from cAMP signaling. We sought to determine how arrestin2 binds to LH/CG It, if binding is of high affinity, and if the receptor also binds arrestin3. Desensitization of intact LH/CG R was equally sensitive to ectopic constructs of arrestin2 that bind other G protein-coupled receptors (GPCRs) either in a phosphorylation-independent or -dependent manner. Intact LH/CG R was not desensitized by ectopic arrestin3 constructs. Surface plasmon resonance studies showed that arrestin2 bound a synthetic third intracellular (3i) LH/CG R loop peptide with picomolar affinity; arrestin3 bound with millimolar affinity. To determine whether Asp-564 in the 3i loop mimicked the phosphorylated residue of other GPCRs, human embryonic kidney (HEK) cells were transfected with wild-type (WT) and D564G LH/CG R, An agonist-stimulated ARF6-dependent arrestin2 undocking pathway to drive desensitization of WT receptor was recapitulated in HEK cell membranes, and ectopic arrestin2 promoted desensitization of WT LH/CG R. However, D564G LH/CG R in HEK cells was not desensitized, and synthetic 3i D564G peptide did not bind arrestin2. Synthetic 3i loop peptides containing D564E, D564V, or D564N also did not bind arrestin2. We conclude that the ARF6-mediated mechanism to release a pool of membrane-delimited arrestin to bind GPCRs may be a widespread mechanism to deliver arrestin to GPCRs for receptor desensitization. Unlike other GPCRs that additionally require receptor phosphorylation, LH/CG R activation is sufficient to expose a conformation in which Asp-564 in the 3i loop confers high affinity binding selectively to arrestin2. C1 Northwestern Univ, Sch Med, Dept Cell & Mol Biol, Chicago, IL 60611 USA. Northwestern Univ, Sch Med, Dept Mol Pharmacol & Biol Chem, Chicago, IL 60611 USA. Northwestern Univ, Sch Med, Inst Neurosci, Chicago, IL 60611 USA. Vanderbilt Univ, Sch Med, Dept Pharmacol, Nashville, TN 37232 USA. CNRS, UPR 2356, F-67084 Strasbourg, France. Univ Illinois, Coll Med, Dept Obstet & Gynecol, Chicago, IL 60612 USA. NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Hunzicker-Dunn, M (reprint author), Northwestern Univ, Sch Med, Dept Cell & Mol Biol, 303 E Chicago Ave, Chicago, IL 60611 USA. RI Hamm, Heidi/G-2374-2014; Gurevich, Vsevolod/A-3236-2008; Dietz-Bader, Marie-France/O-2098-2016 OI Hamm, Heidi/0000-0001-5437-5287; Gurevich, Vsevolod/0000-0002-3950-5351; FU NEI NIH HHS [EY 06062, R01 EY 10291, R01 EY 11500]; NICHD NIH HHS [R01 HD 38060]; NIGMS NIH HHS [GM 63097] NR 73 TC 38 Z9 39 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 MAY 17 PY 2002 VL 277 IS 20 BP 17916 EP 17927 DI 10.1074/jbc.M110479200 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100068 PM 11867621 ER PT J AU Pedersen, LC Petrotchenko, E Shevtsov, S Negishi, M AF Pedersen, LC Petrotchenko, E Shevtsov, S Negishi, M TI Crystal structure of the human estrogen sulfotransferase-PAPS complex - Evidence for catalytic role of Ser(137) in the sulfuryl transfer reaction SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SITE-DIRECTED MUTAGENESIS; CYTOSOLIC SULFOTRANSFERASES; FLAVONOL 3-SULFOTRANSFERASE; SUBSTRATE-SPECIFICITY; MUTATIONAL ANALYSIS; IDENTIFICATION; BINDING; DOMAIN AB Estrogen sulfotransferase (EST) transfers the sulfate group from T-phosphoadenosine 5'-phosphosulfate (PAPS) to estrogenic steroids. Here we report the crystal structure of human EST (hEST) in the context of the V269E mutant-PAPS complex, which is the first structure containing the active sulfate donor for any sulfotransferase. Superimposing this structure with the crystal structure of hEST in complex with the donor product 3'-phosphoadenosine 5'-phosphate (PAP) and the acceptor substrate 17beta-estradiol, the ternary structure with the PAPS and estradiol molecule, is modeled. These structures have now provided a more complete view of the S(N)2-like in-line displacement reaction catalyzed by sulfotransferases. In the PAPS-bound structure, the side chain nitrogen of the catalytic Lys(47) interacts with the side chain hydroxyl of Ser(137) and not with the bridging oxygen between the 5'-phosphate and sulfate groups of the PAPS molecule as is seen in the PAP-bound structures. This conformational change of the side chain nitrogen indicates that the interaction of Lys(47) with Ser(137) may regulate PAPS hydrolysis in the absences of an acceptor substrate. Supporting the structural data, the mutations of Ser(137) to cysteine and alanine decrease gradually k(cat) for PAPS hydrolysis and transfer activity. Thus, Ser(137) appears to play an important role in regulating the side chain interaction of Lys(47) with the bridging oxygen between the 5'-phosphate and the sulfate of PAPS. C1 NIEHS, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Negishi, M (reprint author), NIEHS, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. NR 24 TC 69 Z9 73 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 MAY 17 PY 2002 VL 277 IS 20 BP 17928 EP 17932 DI 10.1074/jbc.M111651200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100069 PM 11884392 ER PT J AU Shakarian, AM Joshi, MB Ghedin, E Dwyer, DM AF Shakarian, AM Joshi, MB Ghedin, E Dwyer, DM TI Molecular dissection of the functional domains of a unique, tartrate-resistant, surface membrane acid phosphatase in the primitive human pathogen Leishmania donovani SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROMASTIGOTES; AMASTIGOTES; MEXICANA; CLONING; PROTEIN; GENES AB The primitive trypanosomatid pathogen of humans, Leishmania donovani, constitutively expresses a unique externally oriented, tartrate-resistant, acid phosphatase on its surface membrane. This is of interest because these organisms are obligate intracellular protozoan parasites that reside and multiply within the hydrolytic milieu of mammalian macrophage phago-lysosomes. Here we report the identification of the gene encoding this novel L. donovani enzyme. In addition, we characterized its structure, demonstrated its constitutive expression in both parasite developmental forms, and determined the cell surface membrane localization of its translated protein product. Further, we used a variety of green fluorescent protein chimeric constructs as reporters in a homologous leishmanial expression system to dissect the functional domains of this unique, tartrate-resistant, surface membrane enzyme. C1 NIAID, Cell Biol Sect, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. Salva Regina Univ, Dept Biol & Biomed Sci, Newport, RI 02840 USA. Inst Genom Res, Rockville, MD 20850 USA. RP Dwyer, DM (reprint author), NIAID, Cell Biol Sect, Parasit Dis Lab, NIH, Bldg 4,Rm 126,4 Ctr Dr,MSC-0425, Bethesda, MD 20892 USA. NR 28 TC 22 Z9 22 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 17 PY 2002 VL 277 IS 20 BP 17994 EP 18001 DI 10.1074/jbc.M200114200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100077 PM 11882658 ER PT J AU Hillion, J Canals, M Torvinen, M Casado, V Scott, R Terasmaa, A Hansson, A Watson, S Olah, ME Mallol, J Canela, EI Zoli, M Agnati, LF Ibanez, CF Lluis, C Franco, R Ferre, S Fuxe, K AF Hillion, J Canals, M Torvinen, M Casado, V Scott, R Terasmaa, A Hansson, A Watson, S Olah, ME Mallol, J Canela, EI Zoli, M Agnati, LF Ibanez, CF Lluis, C Franco, R Ferre, S Fuxe, K TI Coaggregation, cointernalization, and codesensitization of adenosine A(2A) receptors and dopamine D-2 receptors SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID COTRANSFECTED FIBROBLAST CELLS; VOLUME TRANSMISSION; BASAL GANGLIA; A(1); RAT; DESENSITIZATION; DYSKINESIA; STRIATUM; IDENTIFICATION; SCHIZOPHRENIA AB Antagonistic and reciprocal interactions are known to exist between adenosine and dopamine receptors in the striatum. In the present study, double immunofluorescence experiments with confocal laser microscopy showed a high degree of colocalization of adenosine A A receptors (A(2A)R) and dopamine D-2 receptors (D2R) in cell membranes of SH-SY5Y human neuroblastoma cells stably transfected with human D2R and in cultured striatal cells. A(2A)R/D2R heteromeric complexes were demonstrated in coimmunoprecipitation experiments in membrane preparations from D2R-transfected SH-SY5Y cells and from mouse fibroblast Ltk(-) cells stably transfected with human D2R (long form) and transiently cotransfected with the A(2A)R double-tagged with hemagglutinin. Long term exposure to A(2A)R and D2R agonists in D2R-cotransfected SH-SY5Y cells resulted in coaggregation, cointernalization and codesensitization of A(2A)R and D2R. These results give a molecular basis for adenosine-dopamine antagonism at the membrane level and have implications for treatment of Parkinson's disease and schizophrenia, in which D2R are involved. C1 Karolinska Inst, Dept Neurosci, S-17177 Stockholm, Sweden. Univ Barcelona, Dept Biochem & Mol Biol, E-08028 Barcelona, Spain. Univ Michigan, Mental Hlth Inst, Ann Arbor, MI 48109 USA. Duke Univ, Med Ctr, Dept Med, Durham, NC 27710 USA. Univ Modena, Dept Biomed Sci, I-41100 Modena, Italy. NIDA, Baltimore, MD 21224 USA. RP Hillion, J (reprint author), NINCDS, Stroke Branch, NIH, Bethesda, MD 20892 USA. RI Canals, Meritxell/A-6422-2013; Ferre, Sergi/K-6115-2014; Franco, Rafael/C-3694-2015; Terasmaa, Anton/I-3312-2015; Zoli, Michele/C-8899-2016; Casado, Vicent/K-1660-2014; Canela, Enric I./M-8726-2013; OI Ferre, Sergi/0000-0002-1747-1779; Franco, Rafael/0000-0003-2549-4919; Terasmaa, Anton/0000-0002-5139-1764; Zoli, Michele/0000-0002-8034-2849; Canela, Enric I./0000-0003-4992-7440; Canals, Meritxell/0000-0002-7942-5006; Casado, Vicent/0000-0002-1764-3825 NR 35 TC 314 Z9 319 U1 0 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 17 PY 2002 VL 277 IS 20 BP 18091 EP 18097 DI 10.1074/jbc.M107731200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100089 PM 11872740 ER PT J AU Kanduri, C Fitzpatrick, G Mukhopadhyay, R Kanduri, M Lobanenkov, V Higgins, M Ohlsson, R AF Kanduri, C Fitzpatrick, G Mukhopadhyay, R Kanduri, M Lobanenkov, V Higgins, M Ohlsson, R TI A differentially methylated imprinting control region within the Kcnq1 locus harbors a methylation-sensitive chromatin insulator SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BECKWITH-WIEDEMANN-SYNDROME; ENHANCER-BLOCKING ACTIVITY; GENE; CTCF; EXPRESSION; DISRUPTION; TRANSCRIPT; ELEMENTS; KVLQT1; H19 AB The mechanisms underlying the phenomenon of genomic imprinting remain poorly understood. In one instance, a differentially methylated imprinting control region (ICR) at the H19 locus has been shown to involve a methylation-sensitive chromatin insulator function that apparently partitions the neighboring Igf2 and H19 genes in different expression domains in a parent of origin-dependent manner. It is not known, however, if this mechanism is unique to the Igf2/H19 locus or if insulator function is a common feature in the regulation of imprinted genes. To address this question, we have studied an ICR in the Kcnq1 locus that regulates long range repression on the paternally derived p57Kip2 and Kcnq1 alleles in an imprinting domain that includes Igf2 and H19. We show that this ICR appears to possess a unidirectional chromatin insulator function in somatic cells of both mesodermal and endodermal origins. Moreover, we document that CpG methylation regulates this insulator function suggesting that a methylation-sensitive chromatin insulator is a common theme in the phenomenon of genomic imprinting. C1 Uppsala Univ, Evolut Biol Ctr, Dept Genet & Dev, S-75236 Uppsala, Sweden. Roswell Pk Canc Inst, Dept Canc Genet, Buffalo, NY 14263 USA. NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. RP Kanduri, C (reprint author), Uppsala Univ, Evolut Biol Ctr, Dept Genet & Dev, Norbyvagen 18A, S-75236 Uppsala, Sweden. OI Lobanenkov, Victor/0000-0001-6665-3635 NR 17 TC 68 Z9 72 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 MAY 17 PY 2002 VL 277 IS 20 BP 18106 EP 18110 DI 10.1074/jbc.M200031200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 553PR UT WOS:000175685100091 PM 11877438 ER PT J AU Dolensky, B Kirk, KL AF Dolensky, B Kirk, KL TI New building blocks for fluorinated bioimidazole derivatives II: Preparation of beta-fluorourocanic acids SO JOURNAL OF ORGANIC CHEMISTRY LA English DT Article ID BROMOFLUORINATION; IMIDAZOLE; INHIBITORS; UROCANASE; ALCOHOLS; REAGENT; ESTERS AB Replacement of vinyl hydrogen with fluorine is based on addition of an FBr equivalent to a double bond followed by HBr elimination. This sequence has been adapted to prepare 3-fluoro-3-imidazolyl-propenoic acids (beta-fluorourocanic acids), and the related fluorinated imidazolyl propenals and prop-2-en-1-ols, from urocanic acid. Tritylation of the imidazole nitrogen was necessary for successful addition of "FBr" to the double bond, and prior reduction of the carboxyl group to the alcohol was required to provide the desired chemoselectivity in the elimination of HX. Reoxidation and deprotection produced the fluorinated urocanic acids. C1 NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Kirk, KL (reprint author), NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NR 40 TC 14 Z9 14 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-3263 J9 J ORG CHEM JI J. Org. Chem. PD MAY 17 PY 2002 VL 67 IS 10 BP 3468 EP 3473 DI 10.1021/jo0200419 PG 6 WC Chemistry, Organic SC Chemistry GA 552DB UT WOS:000175602300040 PM 12003561 ER PT J AU Diaz-Cabiale, Z Vivo, M Del Arco, A O'Connor, WT Harte, MK Muller, CE Martinez, E Popoli, P Fuxe, K Ferre, S AF Diaz-Cabiale, Z Vivo, M Del Arco, A O'Connor, WT Harte, MK Muller, CE Martinez, E Popoli, P Fuxe, K Ferre, S TI Metabotropic glutamate mGlu5 receptor-mediated modulation of the ventral striopallidal GABA pathway in rats. Interactions with adenosine A(2A) and dopamine D-2 receptors SO NEUROSCIENCE LETTERS LA English DT Article DE adenosine A(2A) receptor; metabotropic mGlu5 receptor; dopamine D-2 receptor; nucleus accumbens; ventral pallidum; gamma-aminobutyric acid; microdialysis; phencyclidine ID 6-HYDROXYDOPAMINE-LESIONED RATS; BINDING CHARACTERISTICS; BASAL GANGLIA; MESSENGER-RNA; STRIATUM; AGONIST; SCHIZOPHRENIA; EXPRESSION; MECHANISM; SYSTEMS AB Interactions between subtypes of dopamine, glutamate and adenosine receptors seem to play an important integrative role in the function of striatal gamma-aminobutyric acid (GABA)ergic efferent neurons. Recent behavioral and biochemical studies suggest the existence of specific interactions between adenosine A(2A) receptors (A2AR), dopamine D-2 receptors (D2R) and the group I metabotropic mGlu5 receptors (mGlu5R) in the dorsal striatum. The dual-probe approach in vivo microdialysis technique in freely moving rats was used to study the role of mGlu5R/A2AR/D2R interactions in the modulation of the ventral striopallidal GABA pathway. Perfusion of a selective mGlu5R agonist (CHPG) in the nucleus accumbens facilitated GABA release in the ipsilateral ventral pallidum. This effect was strongly potentiated by coperfusion with the A2AR agonist CGS 21680. Co-perfusion with the D2R agonist quinpirole counteracted the increase in pallidal GABA levels induced by CGS 21680 and by CGS 21680 plus CHPG. These results demonstrate that mGlu5R/A2AR/D2R interactions play an important modulatory role in the function of the ventral striopallidal GABA pathway, which might have implications for the treatment of schizophrenia and drug addiction. (C) 2002 Published by Elsevier Science Ireland Ltd. C1 NIDA, Behav Neurosci Branch, NIH, Intramural Res Program, Baltimore, MD 21224 USA. Ist Super Sanita, Dept Pharmacol, I-00161 Rome, Italy. Univ Bonn, Inst Pharmaceut, D-53115 Bonn, Germany. Univ Coll Dublin, Dept Human Anat & Physiol, Conway Inst Biomol & Biomed Res, Dublin 2, Ireland. CSIC, Dept Neurochem, IIBB, IDIBAPS, ES-08034 Barcelona, Spain. Karolinska Inst, Dept Neurosci, S-17177 Stockholm, Sweden. RP Ferre, S (reprint author), NIDA, Behav Neurosci Branch, NIH, Intramural Res Program, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Popoli, Patrizia/B-5397-2008; Ferre, Sergi/K-6115-2014; MARTINEZ, EMILI/K-8009-2014; Muller, Christa/C-7748-2014; OI Ferre, Sergi/0000-0002-1747-1779; MARTINEZ, EMILI/0000-0002-3624-6489; Muller, Christa/0000-0002-0013-6624; Diaz-Cabiale, Zaida/0000-0002-1582-2088; O'Connor, William/0000-0003-0082-1119; Harte, Michael/0000-0002-4702-4785; Diaz Cabiale, Maria Zaida/0000-0003-4613-9430 NR 20 TC 86 Z9 88 U1 0 U2 6 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 MAY 17 PY 2002 VL 324 IS 2 BP 154 EP 158 AR PII S0304-3940(02)00179-9 DI 10.1016/S0304-3940(02)00179-9 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 555KK UT WOS:000175793100017 PM 11988350 ER PT J AU Storz, G AF Storz, G TI An expanding universe of noncoding RNAs SO SCIENCE LA English DT Editorial Material ID SIGNAL RECOGNITION PARTICLE; SMALL NUCLEOLAR RNAS; ESCHERICHIA-COLI; CAENORHABDITIS-ELEGANS; COMPARATIVE GENOMICS; IDENTIFICATION; GENES; DATABASE; TRANSCRIPTION; WORLD AB Noncoding RNAs (ncRNAs) have been found to have roles in a great variety of processes, including transcriptional regulation, chromosome replication, RNA processing and modification, messenger RNA stability and translation, and even protein degradation and translocation. Recent studies indicate that ncRNAs are far more abundant and important than initially imagined. These findings raise several fundamental questions: How many ncRNAs are encoded by a genome? Given the absence of a diagnostic open reading frame, how can these genes be identified? How can all the functions of ncRNAs be elucidated? C1 NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Storz, G (reprint author), NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. OI Storz, Gisela/0000-0001-6698-1241 NR 48 TC 347 Z9 379 U1 3 U2 22 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 MAY 17 PY 2002 VL 296 IS 5571 BP 1260 EP 1263 DI 10.1126/science.1072249 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 554BL UT WOS:000175713000038 PM 12016301 ER PT J AU Sun, Y Olson, R Horning, M Armstrong, N Mayer, M Gouaux, E AF Sun, Y Olson, R Horning, M Armstrong, N Mayer, M Gouaux, E TI Mechanism of glutamate receptor desensitization SO NATURE LA English DT Article ID LIGAND-BINDING CORE; AMPA RECEPTOR; ION CHANNELS; TETRAMERIC STRUCTURE; HIPPOCAMPAL-NEURONS; CRYSTAL-STRUCTURES; CYCLOTHIAZIDE; KAINATE; CELLS; SELECTIVITY AB Ligand-gated ion channels transduce chemical signals into electrical impulses by opening a transmembrane pore in response to binding one or more neurotransmitter molecules. After activation, many ligand-gated ion channels enter a desensitized state in which the neurotransmitter remains bound but the ion channel is closed. Although receptor desensitization is crucial to the functioning of many ligand-gated ion channels in vivo, the molecular basis of this important process has until now defied analysis. Using the GluR2 AMPA-sensitive glutamate receptor, we show here that the ligand-binding cores form dimers and that stabilization of the intradimer interface by either mutations or allosteric modulators reduces desensitization. Perturbations that destabilize the interface enhance desensitization. Receptor activation involves conformational changes within each subunit that result in an increase in the separation of portions of the receptor that are linked to the ion channel. Our analysis defines the dimer interface in the resting and activated state, indicates how ligand binding is coupled to gating, and suggests modes of dimer-dimer interaction in the assembled tetramer. Desensitization occurs through rearrangement of the dimer interface, which disengages the agonist-induced conformational change in the ligand-binding core from the ion channel gate. C1 Columbia Univ, Dept Biochem & Mol Biophys, New York, NY 10032 USA. Columbia Univ, Howard Hughes Med Inst, New York, NY 10032 USA. NICHHD, Lab Cellular & Mol Neurophysiol, NIH, Bethesda, MD 20892 USA. RP Gouaux, E (reprint author), Columbia Univ, Dept Biochem & Mol Biophys, 650 W 168th St, New York, NY 10032 USA. RI Mayer, Mark/H-5500-2013 NR 46 TC 483 Z9 492 U1 4 U2 36 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD MAY 16 PY 2002 VL 417 IS 6886 BP 245 EP 253 DI 10.1038/417245a PG 9 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 551YT UT WOS:000175592100037 PM 12015593 ER PT J AU Jayaram, H Taraporewala, Z Patton, JT Prasad, BVV AF Jayaram, H Taraporewala, Z Patton, JT Prasad, BVV TI Rotavirus protein involved in genome replication and packaging exhibits a HIT-like fold SO NATURE LA English DT Article ID TEMPERATURE-SENSITIVE MUTANTS; SINGLE-STRANDED RNA; NONSTRUCTURAL PROTEIN; BLUETONGUE VIRUS; BINDING-PROTEINS; NSP2; MULTIMERS; LOCALIZATION; CORE; NS2 AB Rotavirus, the major cause of life-threatening infantile gastroenteritis, is a member of the Reoviridae(1). Although the structures of rotavirus 2 and other members of the Reoviridae(3,4) have been extensively studied, little is known about the structures of virus-encoded non-structural proteins that are essential for genome replication and packaging. The non-structural protein NSP2 of rotavirus, which exhibits nucleoside triphosphatase, single-stranded RNA binding(5), and nucleic-acid helix-destabilizing(6) activities, is a major component of viral replicase complexes(7,8). We present here the X-ray structure of the functional octamer(9) of NSP2 determined to a resolution of 2.6 Angstrom. The NSP2 monomer has two distinct domains. The amino-terminal domain has a new fold. The carboxy-terminal domain resembles the ubiquitous cellular histidine triad (HIT) group of nucleotidyl hydrolases(10). This structural similarity suggests that the nucleotide-binding site is located inside the cleft between the two domains. Prominent grooves that run diagonally across the doughnut-shaped octamer are probable locations for RNA binding. Several RNA binding sites, resulting from the quaternary organization of NSP2 monomers, may be required for the helix destabilizing activity of NSP2 and its function during genome replication and packaging. C1 Baylor Coll Med, Verna & Marrs McLean Dept Biochem & Mol Biol, Program Struct & Computat Biol & Mol Biophys, Houston, TX 77030 USA. NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Prasad, BVV (reprint author), Baylor Coll Med, Verna & Marrs McLean Dept Biochem & Mol Biol, Program Struct & Computat Biol & Mol Biophys, Houston, TX 77030 USA. RI Patton, John/P-1390-2014 NR 30 TC 62 Z9 70 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD MAY 16 PY 2002 VL 417 IS 6886 BP 311 EP 315 DI 10.1038/417311a PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 551YT UT WOS:000175592100053 PM 12015608 ER PT J AU Sunesen, M Stevnsner, T Brosh, RM Dianov, GL Bohr, VA AF Sunesen, M Stevnsner, T Brosh, RM Dianov, GL Bohr, VA TI Global genome repair of 8-oxoG in hamster cells requires a functional CSB gene product SO ONCOGENE LA English DT Article DE oxidative DNA damage; repair; DNA instability; Cockayne syndrome ID BASE EXCISION-REPAIR; RNA-POLYMERASE-II; OXIDATIVE DNA-DAMAGE; TRANSCRIPTION-COUPLED REPAIR; GROUP-B PROTEIN; COCKAYNE-SYNDROME PATIENTS; XERODERMA-PIGMENTOSUM; MITOCHONDRIAL REPAIR; ELONGATION COMPLEXES; PUTATIVE HELICASES AB Cockayne syndrome (CS) is an autosomal recessive human disease characterized by UV-sensitivity as well as neurological and developmental abnormalities. Two complementation groups have been established, designated CS-A and CS-B. Traditionally, CSA and CSB have been ascribed a function in the transcription-coupled repair (TCR) pathway of nucleotide excision repair (NER) that efficiently removes bulky lesions from the transcribed strand of RNA polymerase 11 transcribed genes. To assess the role of the CSB protein in the repair of the highly mutagenic base lesion 7,8-dihydro-8-oxoguanine (8-oxoG), we have investigated the removal of this lesion using an in vitro incision approach with cell extracts as well as an in vivo approach with a modified protocol of the gene-specific repair assay, which allows the measurement of base lesion repair in intragenomic sequences. Our results demonstrate that the integrity of the CSB protein is pivotal for processes leading to incision at the site of 8-oxoG and that the global genome repair (GGR) of this lesion requires a functional CSB gene product in vivo. C1 NIA, Lab Mol Gerontol, NIH, Baltimore, MD 21224 USA. Aarhus Univ, Dept Biol Mol & Struct, DK-8000 Aarhus C, Denmark. RP Bohr, VA (reprint author), NIA, Lab Mol Gerontol, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 57 TC 39 Z9 40 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 16 PY 2002 VL 21 IS 22 BP 3571 EP 3578 DI 10.1038/sj/onc/1205443 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 555KR UT WOS:000175793700008 PM 12032859 ER PT J AU Talbot, LA Morrell, CH Metter, EJ Fleg, JL AF Talbot, LA Morrell, CH Metter, EJ Fleg, JL TI Comparison of cardiorespiratory fitness versus leisure time physical activity as predictors of coronary events in men aged <= 65 years and > 65 years SO AMERICAN JOURNAL OF CARDIOLOGY LA English DT Article ID HONOLULU-HEART-PROGRAM; CARDIOVASCULAR-DISEASE MORTALITY; HEALTHY-MEN; ELDERLY MEN; FOLLOW-UP; RISK; WALKING; ATTACK; WOMEN; DEATH AB Both high peak oxygen consumption (VO2) and high levels of leisure time physical activity (LTPA) are associated with a reduced risk of cardiovascular morbidity and mortality. We examined the contributions of LTPA and peak VO2 to the risk of coronary events (CEs) in healthy younger (less than or equal to65 years, In = 522) and older (>65 years, n = 167) men from the Baltimore Longitudinal Study of Aging. LTPA derived from self reports of time spent in 97 activities were converted into METs-minutes/24 hours and grouped into high (;2:6 METs), moderate- (4 to 5.9 METs), and low-intensity LTPA (<4 METs). Cardiorespiratory fitness was determined by measuring peak VO2 during a maximal treadmill exercise test. Over a mean follow-up of 13.4 +/- 6.3 years, CEs occurred in 63 men. After accounting for coronary risk factors, proportional-hazards analyses showed a relative CE risk of 0.53 (p <0.0001) for a SD increase in peak VO2 in younger men and 0.61 (p = 0.024) in older men, whereas total LTPA was unrelated to coronary risk in either age group. When the 3 LTPA intensity levels were substituted for total LTPA in the model, peak VO2 remained the only predictor of events in younger men, whereas high-intensity LTPA (RR = 0.39 for tertile 3 vs tertiles I and 2, p 0.016) and peak VO2 (RR = 0.61/SD increase, p 0.024) were of similar importance in older men. Thus, in healthy younger men, higher cardio respiratory fitness but not LTPA predicts a A reduced risk of coronary heart disease, independent of conventional risk factors. For older men, high-intensity LTPA and fitness appear to be of similar importance in reducing coronary risk. (C) 2002 by Excerpta Medica, Inc. C1 Johns Hopkins Univ, Sch Nursing, Baltimore, MD 21205 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. Loyola Coll, Baltimore, MD 21210 USA. RP Talbot, LA (reprint author), Johns Hopkins Univ, Sch Nursing, 525 N Wolfe St, Baltimore, MD 21205 USA. NR 25 TC 56 Z9 58 U1 0 U2 6 PU EXCERPTA MEDICA INC PI NEW YORK PA 650 AVENUE OF THE AMERICAS, NEW YORK, NY 10011 USA SN 0002-9149 J9 AM J CARDIOL JI Am. J. Cardiol. PD MAY 15 PY 2002 VL 89 IS 10 BP 1187 EP 1192 AR PII S0002-9149(02)02302-0 DI 10.1016/S0002-9149(02)02302-0 PG 6 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 552QK UT WOS:000175630900008 PM 12008173 ER PT J AU Philbin, EF Hunsberger, S Garg, R Lader, E Thadani, U McSherry, F Silver, MA AF Philbin, EF Hunsberger, S Garg, R Lader, E Thadani, U McSherry, F Silver, MA CA Digitalis Investigation Grp TI Usefulness of clinical information to distinguish patients with normal from those with low election fractions in heart failure SO AMERICAN JOURNAL OF CARDIOLOGY LA English DT Article ID MANAGEMENT; FEATURES AB The Digitalis Investigation Group (DIG) Trial database was used to determine whether clinical variables used in combination could accurately predict left ejection fraction (EF) in patients with heart failure. Although categorical analysis showed that nearly all of the 38% of patients who had the lowest predicted EF actually had EFs less than or equal to 0.45, this method was unreliable for the remaining 62% of patients. C1 Albany Med Coll, Div Cardiol, Albany, NY 12208 USA. NCI, Bethesda, MD 20892 USA. Eli Lilly & Co, Indianapolis, IN 46285 USA. NYU, Sch Med, New York, NY USA. Univ Oklahoma, Hlth Sci Ctr, Cardiovasc Sect, Oklahoma City, OK USA. VA Med Ctr, Cooperat Studies Program Coordinating Ctr, Perry Point, MD USA. Christ Hosp, Heart Failure Inst, Oak Lawn, IL USA. RP Philbin, EF (reprint author), Albany Med Coll, Div Cardiol, Mail Code 44,47 New Scotland Ave, Albany, NY 12208 USA. FU NHLBI NIH HHS [1Y01-HC00110] NR 12 TC 3 Z9 3 U1 0 U2 0 PU EXCERPTA MEDICA INC PI NEW YORK PA 650 AVENUE OF THE AMERICAS, NEW YORK, NY 10011 USA SN 0002-9149 J9 AM J CARDIOL JI Am. J. Cardiol. PD MAY 15 PY 2002 VL 89 IS 10 BP 1218 EP + AR PII S0002-9149(02)02311-1 DI 10.1016/S0002-9149(02)02311-1 PG 5 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 552QK UT WOS:000175630900017 PM 12008182 ER PT J AU King, TE Toozf, J AF King, TE Toozf, J TI Histopathology and prediction of survival in usual interstitial pneumonia - Reply SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Letter ID IDIOPATHIC PULMONARY FIBROSIS C1 Univ Calif San Francisco, San Francisco, CA 94143 USA. NCI, Rockville, MD USA. RP King, TE (reprint author), Univ Calif San Francisco, San Francisco, CA 94143 USA. NR 6 TC 0 Z9 0 U1 0 U2 1 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1073-449X J9 AM J RESP CRIT CARE JI Am. J. Respir. Crit. Care Med. PD MAY 15 PY 2002 VL 165 IS 10 BP 1451 EP 1451 PG 1 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 553DZ UT WOS:000175661600021 ER PT J AU Borisov, OV Goshe, MB Conrads, TP Rakov, VS Veenstra, TD Smith, RD AF Borisov, OV Goshe, MB Conrads, TP Rakov, VS Veenstra, TD Smith, RD TI Low-energy collision-induced dissociation fragmentation analysis of cysteinyl-modified peptides SO ANALYTICAL CHEMISTRY LA English DT Article ID FLIGHT MASS-SPECTROMETRY; GAS-PHASE CLEAVAGE; QUANTITATIVE-ANALYSIS; PROTEOME ANALYSIS; 2-DIMENSIONAL ELECTROPHORESIS; LIQUID-CHROMATOGRAPHY; GEL-ELECTROPHORESIS; PROTONATED PEPTIDES; SEQUENCE-ANALYSIS; GENE-EXPRESSION AB The development of methods to chemically modify and isolate cysteinyl-residue-containing peptides (Cys-peptides) for LC-MS/MS analysis has generated considerable interest in the field of proteomics. Methods using isotope-coded affinity tags (ICAT) and (+)-biotinyl-iodoacetamidyl-3,6-dioxaoctanediamine (iodoacetyl-PEO-biotin) employ similar Cys-modifying reagents that contain a thiolate-specific biotin group to modify and isolate Cys-containing peptides in conjunction with immobilized avidin. For these strategies to be effective on a proteome-wide level, the presence of the ICAT or acetyl-PEO-biotin tag should not interfere with the efficiency of induced dissociation in MS/MS experiments or with the identification of the modified Cys-peptides by automated database searching algorithms. We have compared the collision-induced dissociation (CID) fragmentation patterns of peptides labeled with iodoacetyl-PEO-biotin and the ICAT reagent to those of the unmodified peptides. CID of Cys-peptides modified with either reagent resulted in the formation of ions attributed to the modified Cys-peptides as well as those unique to the labeling reagent. As demonstrated by analyzing acetyl-PEO-biotin labeled peptides from ribonuclease A and the ICAT-labeled proteome of Deinococcus radiodurans, the presence of these label-specific product ions provides a useful identifier to discern whether a peptide has been modified with the Cys-specific reagent, especially when a number of peptides analyzed using these methods do not contain a modified Cys residue, and to differentiate identical Cys-peptides labeled with either ICAT-d(0) or ICAT-d(s). C1 Pacific NW Natl Lab, Richland, WA 99352 USA. RP Veenstra, TD (reprint author), NCI, Biomed Proteom Program, Analyt Chem Lab, POB B, Frederick, MD 21702 USA. RI Smith, Richard/J-3664-2012 OI Smith, Richard/0000-0002-2381-2349 FU NCI NIH HHS [CA86340, CA93306] NR 26 TC 53 Z9 56 U1 1 U2 8 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 MAY 15 PY 2002 VL 74 IS 10 BP 2284 EP 2292 DI 10.1021/ac010974p PG 9 WC Chemistry, Analytical SC Chemistry GA 553EE UT WOS:000175662100008 PM 12038753 ER PT J AU Miranda, KM Yamada, K Espey, MG Thomas, DD DeGraff, W Mitchell, JB Krishna, MC Colton, CA Wink, DA AF Miranda, KM Yamada, K Espey, MG Thomas, DD DeGraff, W Mitchell, JB Krishna, MC Colton, CA Wink, DA TI Further evidence for distinct reactive intermediates from nitroxyl and peroxynitrite: effects of buffer composition on the chemistry of Angeli's salt and synthetic peroxynitrite SO ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS LA English DT Article DE Angeli's salt; nitroxyl; peroxynitrite; HEPES; bicarbonate; hydrogen peroxide ID NITRIC-OXIDE SYNTHASE; HYDROXYL RADICAL PRODUCTION; HYDROGEN-PEROXIDE; SUPEROXIDE-DISMUTASE; CARBON-DIOXIDE; NMDA RECEPTOR; L-ARGININE; NO; ANION; OXIDATION AB The nitroxyl (HNO) donor Angeli's salt (Na2N2O3; AS) is cytotoxic in vitro, inducing double strand DNA breaks and base oxidation, yet may have pharmacological application in the treatment of cardiovascular disease, The chemical profiles of AS and synthetic peroxynitrite (ONOO-) in aerobic solution were recently compared, and AS was found to form a distinct reactive intermediate. However, similarities in the chemical behavior of the reactive nitrogen oxide species (RNOS) were apparent under certain conditions. Buffer composition was found to have a significant and unexpected impact on the observed chemistry of RNOS, and varied buffer conditions were utilized to further distinguish the chemical profiles elicited by the RNOS donors AS and synthetic ONOO-. Addition of HEPES to the assay buffer significantly quenched oxidation of dihydrorhodamine (DHR), hydroxylation of benzoic acid (BA), and DNA damage by both AS and ONOO-, and oxidation and nitration of hydroxyphenylacetic acid by ONOO-. Additionally, H2O2, was produced in a concentration-dependent manner from the interaction of HEPES with both the donor intermediates. Interestingly, clonogenic survival was not affected by HEPES, indicating that H2O2 is not a contributing factor to in vitro cytotoxicity of AS. Variation in RNOS reactivity was dramatic with significantly higher relative affinity for the AS intermediate toward DHR, BA, DNA, and HEPES and increased production of H2O2. Further, AS reacted to a significantly greater extent with the unprotonated amine form of HEPES while the interaction of ONOO- with HEPES was pH-independent. Addition of bicarbonate only altered ONOO- chemistry. This study emphasizes the importance of buffer composition on chemical outcome and thus on interpretation and provides further evidence that ONOO- is not an intermediate formed between the reaction of O-2 and HNO produced by AS. (C) 2002 Elsevier Science (USA). All rights reserved. C1 NCI, Radiat Biol Branch, NIH, Bethesda, MD 20892 USA. Duke Univ, Med Ctr, Div Neurol, Durham, NC 27710 USA. RP Miranda, KM (reprint author), NCI, Radiat Biol Branch, NIH, Bldg 10,Room B3-B69, Bethesda, MD 20892 USA. RI Miranda, Katrina/B-7823-2009; Yamada, Ken-ichi/E-6318-2012 NR 38 TC 56 Z9 56 U1 0 U2 6 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-9861 J9 ARCH BIOCHEM BIOPHYS JI Arch. Biochem. Biophys. PD MAY 15 PY 2002 VL 401 IS 2 BP 134 EP 144 AR PII S0003-9861(02)00031-0 DI 10.1016/S0003-9861(02)00031-0 PG 11 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 557LU UT WOS:000175910700002 PM 12054463 ER PT J AU Hamid, EH Hyde, TM Egan, MF Wolf, SS Herman, MM Nemeroff, CB Kleinman, JE AF Hamid, EH Hyde, TM Egan, MF Wolf, SS Herman, MM Nemeroff, CB Kleinman, JE TI Neurotensin receptor binding abnormalities in the entorhinal cortex in schizophrenia and affective disorders SO BIOLOGICAL PSYCHIATRY LA English DT Article DE neurotensin; schizophrenia; human brain; entorhinal cortex; hippocampus ID AUTORADIOGRAPHIC CHARACTERIZATION; SUBSTANTIA-NIGRA; REGION; BRAIN; GENE; ORGANIZATION; ASSOCIATION; HYPOTHESIS; SYSTEM; MONKEY AB Background: Convergent evidence from in vivo and in vitro studies of schizophrenia have implicated the mesial temporal lobe as a primary site of pathological change in this disorder. We have previously reported decreased neurotensin receptor density in layer II of the intermediate entorhinal cortex (ERC) in schizophrenia, a finding seen elsewhere but not seen in more caudal ERC. Methods: To study neuroanatomic and diagnostic specificity, we measured the density of neurotensin receptors in the intermediate and caudal ERC and hippocampal formation of schizophrenic, affective disorder control subjects, and normal control subjects. Slide-based radioligand binding was used to perform these studies. Results: Not only schizophrenic but also affective disorder subjects had decreased neurotensin receptor density in layer II of the intermediate ERC Affective disorder subjects had significantly decreased neurotensin receptor density in layers V/VI of the intermediate ERC, and schizophrenic subjects trended in the same direction. Conclusions: These findings demonstrate region-specific changes in neurotensin receptor binding levels in the mesial temporal lobe; however, there is no clear diagnostic specificity for these changes, because they were seen to varying degrees in both schizophrenia and affective disorders. (C) 2002 Society of Biological Psychiatry. C1 NIMH, IRP, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Dept Psychiat & Behav Sci, Atlanta, GA 30322 USA. RP Kleinman, JE (reprint author), NIMH, IRP, Clin Brain Disorders Branch, NIH, 9000 Rockville Pike,MSC1379,Bldg 10,Room 4S237A, Bethesda, MD 20892 USA. NR 24 TC 16 Z9 16 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD MAY 15 PY 2002 VL 51 IS 10 BP 795 EP 800 AR PII S0006-3223(01)01325-7 DI 10.1016/S0006-3223(01)01325-7 PG 6 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 550LF UT WOS:000175504000003 PM 12007453 ER PT J AU Francischetti, IMB Valenzuela, JG Andersen, JF Mather, TN Ribeiro, JMC AF Francischetti, IMB Valenzuela, JG Andersen, JF Mather, TN Ribeiro, JMC TI Ixolaris, a novel recombinant tissue factor pathway inhibitor (TFPI) from the salivary gland of the tick, Ixodes scapularis: Identification of factor X and factor Xa as scaffolds for the inhibition of factor VIIa/tissue factor complex SO BLOOD LA English DT Article ID COAGULATION FACTOR-XA; ANTICOAGULANT PROTEIN C2; FACTOR-VIIA; BLOOD-COAGULATION; EXTRINSIC PATHWAY; FULL-LENGTH; MECHANISM; RESIDUES; SURFACE; DOMAINS AB Saliva of the hard tick and Lyme disease vector, Ixodes scapularis, has a repertoire of compounds that counteract host defenses. Following sequencing of an I scapularis salivary gland complementary DNA (cDNA) library, a clone with sequence homology to tissue factor pathway inhibitor (TFPI) was identified. This cDNA codes for a mature protein, herein called Ixolarls, with 140 amino acids containing 10 cysteines and 2 Kunitz-like domains. Recombinant Ixolarls was expressed in insect cells and shown to inhibit factor VIIa (FVIIa)/ tissue factor (TF)-induced factor X (FX) activation with an inhibitory concentration of 50% (IC50) in the picomolar range. In nondenaturing gel, Ixolaris Interacted stoichiometrically with FX and FXa but not FVIIa. Ixolaris behaves as a fast-and tight ligand of the exosites of FXa and T-carboxyglutamic acid domainless FXa (des-Gla-FXa), increasing its amidolytic activity. At high concentration, Ixolaris attenuates the amiclolytic activity of FVIIa/TF; however, in the presence of DEGR-FX or DEGR-FXa (but not des-Gla-DEGR-FXa), Ixolaris becomes a tight inhibitor of FVIIa/TF as assessed by recombinant factor IX (BeneFIX) activation assays. This indicates that FX and FXa are scaffolds for Ixolaris in the Inhibition of FVIIa/TF and Implies that the Gila domain Is necessary for FVIIa/TF/Ixolaris/ FX(a) complex formation. Additionally, we show that Ixolaris blocks FXa generation by endothelial cells expressing TF. Ixolaris may be a useful tool to study the structural features of FVIIa, FX, and FXa, and an alternative anticoagulant In cardiovascular diseases. (C) 2002 by The American Society of Hematology. C1 NIAID, Parasit Dis Lab, Sect Med Ematol, NIH, Bethesda, MD 20892 USA. Univ Rhode Isl, Ctr Vector Borne Dis, Kingston, RI 02881 USA. RP Ribeiro, JMC (reprint author), NIAID, Parasit Dis Lab, Sect Med Ematol, NIH, 4 Ctr Dr,Rm 4-126,MSC 0425, Bethesda, MD 20892 USA. OI Ribeiro, Jose/0000-0002-9107-0818 NR 62 TC 166 Z9 175 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 MAY 15 PY 2002 VL 99 IS 10 BP 3602 EP 3612 DI 10.1182/blood-2001-12-0237 PG 11 WC Hematology SC Hematology GA 550GY UT WOS:000175496300020 PM 11986214 ER PT J AU Kook, H Risitano, AM Zeng, WH Wlodarski, M Lottemann, C Nakamura, R Barrett, J Young, NS Maciejewski, JP AF Kook, H Risitano, AM Zeng, WH Wlodarski, M Lottemann, C Nakamura, R Barrett, J Young, NS Maciejewski, JP TI Changes in T-cell receptor VB repertoire in aplastic anemia: effects of different immunosuppressive regimens SO BLOOD LA English DT Article ID PAROXYSMAL-NOCTURNAL HEMOGLOBINURIA; DONOR LYMPHOCYTE INFUSION; VERSUS-HOST DISEASE; BONE-MARROW; PERIPHERAL-BLOOD; IN-VIVO; CLONAL EXPANSIONS; CDR3 MOTIFS; ANTIGEN; TCR AB We studied the degree and the pattern of skewing of the variable region of beta-chain (VB) T-cell receptor (TCR) repertoire in aplastic anemia (AA) at initial presentation and after immunosuppression using a high-resolution analysis of the TCR VB complementarity-determining region 3 (CDR3). Age-matched healthy individuals and multitransfused patients with nonimmune-mediated hematologic diseases were used as controls. In newly diagnosed AA, the average frequency of CDR3 size distribution deviation indicative of oligoclonal T-cell proliferation was increased (44% 33% vs 9% 9%; P =.0001); AA patients with human leukocyte antigen (HLA)-DR2 and those with expanded paroxysmal nocturnal hemoglobinuria clones showed more skewed VB repertoires. Nonrandom oligoclonal patterns were found for VB6, VB14-16, VB21, VB23, and VB24 subfamilies in more than 50%, and for VB15, VB21, and VB24 in more than 70% of AA patients with HLA-DR2. Patients received immunosuppression with antithymocyte globulin (ATG)/ cyclosporine (CsA) or cyclophosphamide (CTX) with CsA in combination, and their VB repertoire was reanalyzed after treatment. Whereas no significant change in the degree of VB skewing In patients who had received ATG was seen, patients treated with CTX showed a much higher extent of oligoclonality within all VB families, consistent with a profound and long-lasting contraction of the T-cell repertoire. VB analysis did not correlate with the lymphocyte count prior to lymphocytotoxic therapy; however, after therapy the degree of VB skewing was highly reflective of the decrease In lymphocyte numbers, suggesting iatrogenic gaps in the VB repertoire rather than the emergence of clonal dominance. Our data Indicate that multiple specific clones mediate the Immune process In AA. (C) 2002 by The American Society of Hematology. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Maciejewski, JP (reprint author), Taussig Canc Ctr, R40,9500 Euclid Ave, Cleveland, OH 44195 USA. NR 51 TC 43 Z9 46 U1 0 U2 1 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 MAY 15 PY 2002 VL 99 IS 10 BP 3668 EP 3675 DI 10.1182/blood.V99.10.3668 PG 8 WC Hematology SC Hematology GA 550GY UT WOS:000175496300028 PM 11986222 ER PT J AU van der Kolk, DM Vellenga, E Scheffer, GL Muller, M Bates, SE Scheper, RJ de Vries, EGE AF van der Kolk, DM Vellenga, E Scheffer, GL Muller, M Bates, SE Scheper, RJ de Vries, EGE TI Expression and activity of breast cancer resistance protein (BCRP) in de novo and relapsed acute myeloid leukemia SO BLOOD LA English DT Article ID BONE-MARROW TRANSPLANTATION; MULTIDRUG-RESISTANCE; P-GLYCOPROTEIN; CELL-LINES; MITOXANTRONE-RESISTANT; HALF-TRANSPORTER; OVEREXPRESSION; GENE; MRP1; PHENOTYPE AB Overexpression of the breast cancer resistance protein (BCRP) efflux pump In human cancer cell lines results in resistance to a variety of cytostatic agents. The aim of this study was to analyze BCRP protein expression and activity In acute myeloid leukemia (AML) samples and to determine whether it is up-regulated due to clonal selection at relapse/ refractory disease. BCRP protein expression was measured flow cytometrically with the monoclonal antibodies BXP-34 and BXP-21 in 20 paired samples of de novo and relapsed/refractory AML. BXP-34/immunoglobulin G1 ratios were observed of 1.6 +/- 0.5 (mean +/- SD, range 0.8-2.7) and BXP-21/immunoglobulin G2a ratios of 4.9 +/- 3.0 (range 1.1-14.5) in the patient samples versus 9.8 +/- 6.8 and 6.5 +/- 2.4, respectively, in the MCF-7 cell line. BCRP activity was determined flow cytometrically by measuring mitoxantrone accumulation in absence and presence of the inhibitor fumitremorgin C. Mitoxantrone accumulation, expressed as mean fluorescence intensity (MFI), varied between 44 and 761 MFI (227 +/- 146 MFI) and correlated inversely with BCRP expression (r = -0.58, P <.001). Addition of fumitremorgin C showed a small increase in mitoxantrone accumulation (11 +/- 29 MFI, n = 40) apart from the effect of PSC833 and MK-571. No consistent upregulation of BCRP expression or activity was observed at relapse/refractory disease; some cases showed an increase and other cases a decrease at relapse. Relatively high BCRP expression correlated with immature immunophenotype, as determined by expression of the surface marker CD34 (r = 0.54, P =.001). In conclusion, this study shows that BCRP protein is expressed at low but variable levels in AML, especially in immature CD34(+) cells. BCRP was not consistently up-regulated in relapsed/refractory AML. (C) 2002 by The American Society of Hematology. C1 Univ Groningen Hosp, Dept Internal Med, Div Med Oncol, NL-9713 GZ Groningen, Netherlands. Univ Groningen Hosp, Div Hematol, NL-9713 GZ Groningen, Netherlands. Free Univ Amsterdam, Med Ctr, Dept Pathol, Amsterdam, Netherlands. Univ Wageningen & Res Ctr, Div Nutr Metab & Genom, Wageningen, Netherlands. NCI, Med Branch, NIH, Bethesda, MD 20892 USA. RP de Vries, EGE (reprint author), Univ Groningen Hosp, Dept Internal Med, Div Med Oncol, Hanzeplein 1, NL-9713 GZ Groningen, Netherlands. RI Muller, Michael/B-5795-2008 OI Muller, Michael/0000-0002-5930-9905 NR 41 TC 98 Z9 98 U1 0 U2 1 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 MAY 15 PY 2002 VL 99 IS 10 BP 3763 EP 3770 DI 10.1182/blood.V99.10.3763 PG 8 WC Hematology SC Hematology GA 550GY UT WOS:000175496300040 PM 11986234 ER PT J AU Egan, AF Fabucci, ME Saul, A Kaslow, DC Miller, LH AF Egan, AF Fabucci, ME Saul, A Kaslow, DC Miller, LH TI Aotus New World monkeys: model for studying malaria-induced anemia SO BLOOD LA English DT Article ID MEROZOITE SURFACE PROTEIN-1; P-FALCIPARUM MALARIA; IN-VIVO; CHILDREN; AREA; ERYTHROPHAGOCYTOSIS; DYSERYTHROPOIESIS; CHEMOPROPHYLAXIS; INFECTIONS; MORBIDITY AB Falciparum malaria is a major cause of disease and death in African children and pregnant women, primarily due to severe anemia. We studied anemia In vaccinated Aotus monkeys during a second Infection where the animals were considered to be semi-immune. Most animals had extremely low or undetectable levels of parasitemia; In some, anemia did not develop and reticulocytemia remained unchanged; in others, moderate to severe anemia developed with inappropriately low reticulocytemia indicating bone marrow dysfunction. Bone marrow rapidly responded after parasite clearance. The rapid drop in hematocrit despite extremely low to undetectable parasitemia indicated massive removal of uninfected red blood cells from the circulation that, in the presence of bone marrow dysfunction, led to severe anemia-the problem that occurs in African children. We demonstrate that Aotus monkeys are a nonhuman primate model to gain insight into the pathogenesis of severe anemia in African children. (C) 2002 by The American Society of Hematology. C1 NIAID, Lab Parasit Dis, Malaria Vaccines Sect, NIH, Bethesda, MD USA. NIAID, Lab Parasit Dis, Malaria Vaccine Dev Unit, NIH, Bethesda, MD USA. RP Egan, AF (reprint author), NIH, Fogarty Int Ctr, Rm 31-B2C39 MSC 2220 31 Ctr Dr, Bethesda, MD 20892 USA. RI Saul, Allan/I-6968-2013 OI Saul, Allan/0000-0003-0665-4091 NR 20 TC 40 Z9 40 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 MAY 15 PY 2002 VL 99 IS 10 BP 3863 EP 3866 DI 10.1182/blood.V99.10.3863 PG 4 WC Hematology SC Hematology GA 550GY UT WOS:000175496300057 PM 11986251 ER PT J AU Yu, XB Noguchi, CT AF Yu, XB Noguchi, CT TI Response: Equivalence of mouse and human erythropoietin/erythropoietin receptor signaling SO BLOOD LA English DT Letter ID HUMAN ERYTHROPOIETIN RECEPTOR; GENE; EXPRESSION; TRANSCRIPTION; POLYCYTHEMIA; GATA-1; REGION C1 NIDDK, Lab Chem Biol, Natl Inst Hlth, Bethesda, MD 20892 USA. RP Noguchi, CT (reprint author), NIDDK, Lab Chem Biol, Natl Inst Hlth, 10 Ctr Dr MSC 1822, Bethesda, MD 20892 USA. NR 13 TC 1 Z9 1 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 MAY 15 PY 2002 VL 99 IS 10 BP 3874 EP 3875 PG 2 WC Hematology SC Hematology GA 550GY UT WOS:000175496300066 ER PT J AU Weissfeld, JL Ling, BS Schoen, RE Bresalier, RS Riley, T Prorok, PC AF Weissfeld, JL Ling, BS Schoen, RE Bresalier, RS Riley, T Prorok, PC CA PLCO Project Team TI Adherence to repeat screening flexible sigmoidoscopy in the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial SO CANCER LA English DT Article DE flexible sigmoidoscopy; acceptability; adherence; repeat testing ID PRIMARY-CARE PHYSICIANS; RISK AB BACKGROUND. Acceptance of screening flexible sigmoidoscopy has been poor, in part because of providers' concerns regarding the acceptability of the procedure. In the current prospective study, the authors used adherence to repeat testing to assess the acceptability of screening flexible sigmoidoscopy. METHODS. The current study was a prospective study of a randomized clinical trial drawing volunteers from the community. Subjects included 10,164 Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial participants who were available for follow-up 3 years after undergoing a baseline screening flexible sigmoidoscopy examination. The authors measured adherence and identified those factors that appeared to affect adherence to repeat sigmoidoscopy. RESULTS. Overall, 18.3% of women and 10.0% of men did not undergo a repeat sigmoidoscopy. Among individuals who attended the Year-3 clinic, 10.4% of women and 5.1% of men specifically refused repeat sigmoidoscopy when it was offered (risk of refusal in women compared with men, 2.04; 95% confidence interval, 1.76-2.36). Another factor found to be associated with refusal included a technically inadequate baseline sigmoidoscopy. CONCLUSIONS. Gender and past experiences with sigmoidoscopy may impact adherence to repeat screening. Nonetheless, among research volunteers in a randomized clinical trial of screening, excellent adherence to repeat screening flexible sigmoidoscopy could be achieved. (C) 2002 American Cancer Society. C1 Univ Pittsburgh, Inst Canc, Pittsburgh, PA 15260 USA. Henry Ford Hlth Sci Ctr, Detroit, MI USA. Informat Management Syst Inc, Silver Spring, MD USA. NCI, Bethesda, MD 20892 USA. RP Weissfeld, JL (reprint author), Keystone Bldg,Suite 510,3520 5th Ave, Pittsburgh, PA 15213 USA. NR 22 TC 19 Z9 19 U1 0 U2 0 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0008-543X J9 CANCER JI Cancer PD MAY 15 PY 2002 VL 94 IS 10 BP 2569 EP 2576 DI 10.1002/cncr.10538 PG 8 WC Oncology SC Oncology GA 548JA UT WOS:000175383800007 PM 12173322 ER PT J AU Edwards, BK Howe, HL Ries, LAG Thun, MJ Rosenberg, HM Yancik, R Wingo, PA Jemal, A Feigal, EG AF Edwards, BK Howe, HL Ries, LAG Thun, MJ Rosenberg, HM Yancik, R Wingo, PA Jemal, A Feigal, EG TI Annual report to the nation on the status of cancer, 1973-1999, featuring implications of age and aging on US cancer burden SO CANCER LA English DT Article DE neoplasm; incidence; mortality; aging; race/ethnicity; surveillance; joinpoint ID TREATMENT TRIALS; SPECIAL SECTION; OLDER PATIENTS; TRENDS; PERSPECTIVES; MORTALITY; PROGRESS; ISSUES; RATES AB BACKGROUND. The American Cancer Society, the National Cancer Institute, the North American Association of Central Cancer Registries (NAACCR), the National Institute on Aging (NIA), and the Centers for Disease Control and Prevention, including the National Center for Health Statistics (NCHS) and the National Center for Chronic Disease Prevention and Health Promotion, collaborated to provide an annual update on cancer occurrence and trends in the United States. This year's report contained a special feature focusing on implications of age and aging on the U.S. cancer burden. METHODS, For 1995 through 1999, age-specific rates and age-adjusted rates were calculated for the major cancers using incidence data from the Surveillance, Epidemiology, and End Results Program, the National Program of Cancer Registries, and the NAACCR, and mortality data from NCHS. Joinpoint analysis, a model of joined line segments, was used to examine 1973-1999 trends in incidence and death rates by age for the four most common cancers. Deaths were classified using the eighth, ninth, and tenth revisions of the International Classification of Diseases. Age-adjusted incidence and death rates were standardized to the year 2000 population, which places more emphasis on older persons, in whom cancer rates are higher. RESULTS. Across all ages, overall cancer death rates decreased in men and women from 1993 through 1999, while cancer incidence rates stabilized from 1995 through 1999. Age-specific trends varied by site, sex, and race. For example, breast cancer incidence rates increased in women aged 50-64 years, whereas breast cancer death rates decreased in each age group. However, a major determinant of the future cancer burden is the demographic phenomenon of the aging and increasing size of the U.S. population. The total number of cancer cases can be expected to double by 2050 if current incidence rates remain stable. CONCLUSIONS. Despite the continuing decrease in cancer death rates and stabilization of cancer incidence rates, the overall growth and aging of the U.S. population can be expected to increase the burden of cancer in our nation. (C) 2002 American Cancer Society. C1 NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. N Amer Assoc Cent Canc Registries, Springfield, IL USA. Amer Canc Soc, Epidemiol & Surveillance Res Dept, Atlanta, GA 30329 USA. Ctr Dis Control & Prevent, Natl Ctr Hlth Stat, Div Vital Stat, Hyattsville, MD 20782 USA. NIA, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Natl Ctr Chron Dis Prevent & Hlth Promot, Div Canc Prevent & Control, Atlanta, GA USA. NCI, Div Canc Treatment & Diag, Bethesda, MD 20892 USA. RP Edwards, BK (reprint author), NCI, Div Canc Control & Populat Sci, 6116 Execut Blvd,Suite 504 MSC 8315, Bethesda, MD 20892 USA. NR 65 TC 571 Z9 593 U1 2 U2 13 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0008-543X J9 CANCER JI Cancer PD MAY 15 PY 2002 VL 94 IS 10 BP 2766 EP 2792 DI 10.1002/cncr.10593 PG 27 WC Oncology SC Oncology GA 548JA UT WOS:000175383800033 PM 12173348 ER PT J AU Kim, JS Crooks, H Dracheva, T Nishanian, TG Singh, B Jen, J Waldman, T AF Kim, JS Crooks, H Dracheva, T Nishanian, TG Singh, B Jen, J Waldman, T TI Oncogenic beta-catenin is required for bone morphogenetic protein 4 expression in human cancer cells SO CANCER RESEARCH LA English DT Article ID GENE; BMP; MUTATIONS; PATHWAY AB Somatic cell gene targeting was used to create an isogenic set of human colon cancer cells that differs only in the presence or absence of their endogenous activated beta-catenin oncogene. Affymetrix Genechip expression profiling of parental cells and gene-targeted derivatives identified numerous novel genes whose expression was dependent on the presence of oncogenic beta-catenin. The transforming growth factor-beta family member bone morphogenetic protein 4 (BMP4), whose receptor is mutated in a rare inherited gastrointestinal cancer predisposition syndrome, was the most highly differentially expressed gene. Additional experiments revealed that the oncogenic allele of beta-catenin specifically is absolutely required for BMP4 expression and secretion by human cancer cells and that BMP4 is overexpressed and secreted by human colon cancer cells with mutant adenomatous polyposis coli genes. These data identify the presence of regulatory interactions between the Writ and BMP signaling pathways in cancer pathogenesis, providing an intriguing connection between the sporadic and inherited forms of a common human malignancy. C1 Georgetown Univ, Sch Med, Vincent T Lombardi Canc Res Ctr, Dept Oncol, Washington, DC 20007 USA. Georgetown Univ, Sch Med, Vincent T Lombardi Canc Res Ctr, Dept Pathol, Washington, DC 20007 USA. Georgetown Univ, Sch Med, Vincent T Lombardi Canc Res Ctr, Tumor Biol Training Program, Washington, DC 20007 USA. NCI, Lab Populat Genet, Ctr Canc Res, Bethesda, MD 20892 USA. RP Waldman, T (reprint author), Georgetown Univ, Sch Med, Vincent T Lombardi Canc Res Ctr, Dept Oncol, Res Bldg,Room E304,3970 Reservoir Rd NW, Washington, DC 20007 USA. FU NCI NIH HHS [K01 CA 87828, P30 CA 51008] NR 18 TC 117 Z9 120 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 MAY 15 PY 2002 VL 62 IS 10 BP 2744 EP 2748 PG 5 WC Oncology SC Oncology GA 552ZR UT WOS:000175650900004 PM 12019147 ER PT J AU Hewitt, SC Bocchinfuso, WP Zhai, J Harrell, C Koonce, L Clark, J Myers, P Korach, KS AF Hewitt, SC Bocchinfuso, WP Zhai, J Harrell, C Koonce, L Clark, J Myers, P Korach, KS TI Lack of ductal development in the absence of functional estrogen receptor alpha delays mammary tumor formation induced by transgenic expression of ErbB2/neu SO CANCER RESEARCH LA English DT Article ID HUMAN BREAST-CANCER; GROWTH-FACTOR RECEPTOR; CYCLIN D1; NEU ONCOGENE; MOUSE MODELS; ACTIVATING MUTATIONS; NEU/ERBB-2 ONCOGENE; SIGNAL-TRANSDUCTION; PROTO-ONCOGENE; MICE BEARING AB Expression of the mouse mammary tumor virus (MMTV) neu/erbB2 transgene in mice induces mammary tumors. To examine the effect of removing estrogen receptor alpha (ERalpha) signaling on the ability of an MMTV-neu/erbB2 transgene to induce mammary tumors, the neu transgene was expressed in the ERalpha knockout (alphaERKO) mouse, which lacks functional ERalpha. MMTV-neu females that lacked ERalpha still developed mammary tumors; however, tumor onset was significantly delayed. This study indicates that ERalpha is not required for mammary tumor induction by overexpression of neu/erbB2, but plays a role in the rate of tumor onset. The removal of ovarian steroid by ovariectomy in adults did not alter the onset rate. In contrast, prepubertal ovariectomy, which arrested mammary epithelial development, significantly delayed onset. In addition, manipulations that increase progesterone also accelerate the tumor onset, indicating the slower onset in the alphaERKO is primarily attributable to the anovulatory phenotype resulting in lack of progesterone stimulation and a decreased abundance of target cells in the alphaERKO mammary gland. C1 NIEHS, Receptor Biol Sect, LRDT, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Comparat Med Branch, NIH, Res Triangle Pk, NC 27709 USA. Analyt Sci Inc, Durham, NC 27713 USA. RP Korach, KS (reprint author), NIEHS, Receptor Biol Sect, LRDT, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 58 TC 32 Z9 33 U1 1 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 MAY 15 PY 2002 VL 62 IS 10 BP 2798 EP 2805 PG 8 WC Oncology SC Oncology GA 552ZR UT WOS:000175650900013 PM 12019156 ER PT J AU Sawiris, GP Sherman-Baust, CA Becker, KG Cheadle, C Teichberg, D Morin, PJ AF Sawiris, GP Sherman-Baust, CA Becker, KG Cheadle, C Teichberg, D Morin, PJ TI Development of a highly specialized cDNA array for the study and diagnosis of epithelial ovarian cancer SO CANCER RESEARCH LA English DT Article ID GENE-EXPRESSION PROFILES; SURFACE EPITHELIUM; BREAST-CANCER; FREQUENT LOSS; IGF-II; TUMORS; CELLS; MICROARRAY; CARCINOMAS; PATTERNS AB Ovarian cancer is a major cause of cancer death in women. Unfortunately, the molecular pathways underlying ovarian cancer progression are poorly understood, making the development of novel diagnostic and therapeutic strategies difficult. On the basis of our previous observations obtained from serial analysis of gene expression, we have constructed a specialized cDNA array for the study of ovarian cancer. Small, specialized arrays have several practical advantages and can reveal information that is lost in the "noise" generated by irrelevant genes present in larger arrays. The array, which we named Ovachip, contains 516 cDNAs chosen from our serial analysis of gene expression and cDNA array studies for their relevance to ovarian cancer. The gene expression patterns revealed with the Ovachip are highly reproducible and extremely consistent among the different ovarian specimens tested. This array was extremely sensitive at differentiating ovarian cancer from colon cancer based on expression profiles. The Ovachip revealed clusters of coordinately expressed genes in ovarian cancer. One such cluster, the IGF2 cluster, is particularly striking and includes the insulin-like growth factor II, the cisplatin resistance-associated protein, the checkpoint suppressor I, the cyclin-dependent kinase 6, and a protein tyrosine phosphatase receptor. We also identified a cluster of down-regulated genes that included the cyclin-dependent kinase 7 and cyclin H. Thus, the Ovachip allowed us to identify previously unidentified clusters of differentially expressed genes that may provide new paradigms for molecular pathways important in ovarian malignancies. Because of the relevance of the arrayed genes, the Ovachip may become a powerful tool for investigators in the field of ovarian cancer and may facilitate progress in understanding the etiology of this disease and in its clinical management. C1 NIA, Gerontol Res Ctr, Cellular & Mol Biol Lab, NIH, Baltimore, MD 21224 USA. NIA, Gerontol Res Ctr, DNA Array Unit, NIH, Baltimore, MD 21224 USA. RP Morin, PJ (reprint author), NIA, Gerontol Res Ctr, Cellular & Mol Biol Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Becker, Kevin/0000-0002-6794-6656 NR 29 TC 84 Z9 93 U1 2 U2 4 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 MAY 15 PY 2002 VL 62 IS 10 BP 2923 EP 2928 PG 6 WC Oncology SC Oncology GA 552ZR UT WOS:000175650900030 PM 12019173 ER PT J AU Yu, YL Merlino, G AF Yu, YL Merlino, G TI Constitutive c-Met signaling through a nonautocrine mechanism promotes metastasis in a transgenic transplantation model SO CANCER RESEARCH LA English DT Article ID HEPATOCYTE GROWTH-FACTOR; FACTOR SCATTER FACTOR; RECEPTOR TYROSINE KINASE; FACTOR/SCATTER FACTOR; COLORECTAL-CARCINOMA; LIVER COLONIZATION; MALIGNANT-MELANOMA; TUMOR PROGRESSION; MET/HGF RECEPTOR; INVASIVE GROWTH AB Normal cells are dependent on exogenous, receptor-mediated growth stimulation for cell cycle entry and progression, providing a critical homeostatic mechanism regulating cellular proliferation. In contrast, tumor cells acquire some degree of growth signal autonomy, often through their ability to produce growth factors as well as their receptors (autocrine signaling). Recently, data have begun to emerge implicating heterotypic signaling between diverse cell types within a tumor in the genesis and progression of cancer; however, current experimental approaches in vivo have not adequately addressed this critical relationship. Here we used transgenic mice overexpressing hepatocyte growth factor/scatter factor (HGF/SF), or its growth antagonist NK2, as genetically modified hosts for transplantation of tumor cells expressing their receptor, c-Met, to directly assess the contribution of heterotypic signaling to metastatic colonization. We demonstrate that metastatic potential under nonautocrine signaling conditions (i.e., where tumor cells expressing c-Met are transplanted into transgenic hosts producing HGF/SF) rivaled that observed under conditions of autocrine signaling (i.e., where tumor cells expressing both HGF/SF and c-Met are transplanted into wild-type hosts). HGF/SF and NK2 were not functionally equivalent in vivo. Attenuation of NK2-associated growth inhibition by the presence of an HGF/SF-Met autocrine loop uncovered a shift in metastatic site preference from lung to liver only in NK2-transgenic hosts, a qualitative behavioral alteration likely detectable only through the genetic approach used here. Our data demonstrate that growth factors not intrinsic to malignant cells can have profound effects on metastatic efficiency in vivo and provide experimental support of a role for heterotypic signaling in tumor progression. C1 NCI, Mol Genet Sect, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Merlino, G (reprint author), NCI, Mol Genet Sect, Mol Biol Lab, NIH, Bldg 37,Room 5002, Bethesda, MD 20892 USA. NR 59 TC 41 Z9 43 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 MAY 15 PY 2002 VL 62 IS 10 BP 2951 EP 2956 PG 6 WC Oncology SC Oncology GA 552ZR UT WOS:000175650900034 PM 12019177 ER PT J AU Citri, A Alroy, I Lavi, S Rubin, C Xu, WP Grammatikakis, N Patterson, C Neckers, L Fry, DW Yarden, Y AF Citri, A Alroy, I Lavi, S Rubin, C Xu, WP Grammatikakis, N Patterson, C Neckers, L Fry, DW Yarden, Y TI Drug-induced ubiquitylation and degradation of ErbB receptor tyrosine kinases: implications for cancer therapy SO EMBO JOURNAL LA English DT Article DE geldanamycin; growth factor; protein kinase inhibitors; stress response; ubiquitin ID GROWTH-FACTOR RECEPTOR; HUMAN-BREAST; IRREVERSIBLE INHIBITORS; CHEMOTHERAPEUTIC-AGENTS; MONOCLONAL-ANTIBODIES; HERBIMYCIN-A; C-CBL; GELDANAMYCIN; ONCOGENE; ASSOCIATION AB Overexpression of ErbB-2/HER2 is associated with aggressive human malignancies, and therapeutic strategies targeting the oncoprotein are currently in different stages of clinical application. Tyrosine kinase inhibitors (TKIs) that block the nucleotide-binding site of the kinase are especially effective against tumors. Here we report an unexpected activity of TKIs: along with inhibition of tyrosine phosphorylation, they enhance ubiquitylation and accelerate endocytosis and subsequent intracellular destruction of ErbB-2 molecules. Especially potent is an irreversible TKI (CI-1033) that alkylates a cysteine specific to ErbB receptors. The degradative pathway stimulated by TKIs appears to be chaperone mediated, and is common to the heat shock protein 90 (Hsp90) antagonist geldanamycin and a stress-induced mechanism. In agreement with this conclusion, CI-1033 and geldanamycin additively inhibit tumor cell growth. Based upon a model for drug-induced degradation of ErbB-2, we propose a general strategy for selective destruction of oncoproteins by targeting their interaction with molecular chaperones. C1 Weizmann Inst Sci, Dept Regulat Biol, IL-76100 Rehovot, Israel. NCI, Med Branch, Rockville, MD 20850 USA. Univ N Carolina, Program Mol Cardiol, Chapel Hill, NC USA. Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC USA. Pfizer Global Res & Dev, Dept Canc Res, Ann Arbor, MI 48106 USA. Queens Univ, Dept Microbiol & Immunol, Kingston, ON K7L 3N6, Canada. RP Yarden, Y (reprint author), Weizmann Inst Sci, Dept Regulat Biol, IL-76100 Rehovot, Israel. RI YARDEN, YOSEF/K-1467-2012 FU NCI NIH HHS [CA72981, R01 CA072981, R37 CA072981]; NHLBI NIH HHS [HL03658] NR 32 TC 166 Z9 170 U1 3 U2 14 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD MAY 15 PY 2002 VL 21 IS 10 BP 2407 EP 2417 DI 10.1093/emboj/21.10.2407 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 554TQ UT WOS:000175753600011 PM 12006493 ER PT J AU Sironi, L Mapelli, M Knapp, S De Antoni, A Jeang, KT Musacchio, A AF Sironi, L Mapelli, M Knapp, S De Antoni, A Jeang, KT Musacchio, A TI Crystal structure of the tetrameric Mad1-Mad2 core complex: implications of 'safety belt' binding mechanism for the spindle checkpoint SO EMBO JOURNAL LA English DT Article DE anaphase; Cdc20; Mad1; Mad2; mitotic spindle checkpoint ID ANAPHASE-PROMOTING COMPLEX; ASSEMBLY CHECKPOINT; BUDDING YEAST; UNATTACHED KINETOCHORES; PROTEIN MAD2; CDC20; ASSOCIATION; CELLS; IDENTIFICATION; INHIBITION AB The spindle checkpoint protein Mad1 recruits Mad2 to unattached kinetochores and is essential for Mad2-Cdc20 complex formation in vivo but not in vitro. The crystal structure of the Mad1-Mad2 complex reveals an asymmetric tetramer, with elongated Mad1 monomers parting from a coiled-coil to form two connected sub-complexes with Mad2 The Mad2 C-terminal tails are hinged mobile elements wrapping around the elongated ligands like molecular 'safety belts'. We show that Mad1 is a competitive inhibitor of the Mad2-Cdc20 complex, and propose that the Mad1-Mad2 complex acts as a regulated gate to control Mad2 release for Cdc20 binding. Mad1-Mad2 is strongly stabilized in the tetramer, but a 1:1 Mad1-Mad2 complex slowly releases Mad2 for Cdc20 binding, driven by favourable binding energies. Thus, the rate of Mad2 binding to Cdc20 during checkpoint activation may be regulated by conformational changes that destabilize the tetrameric Mad1-Mad2 assembly to promote Mad2 release. We also show that unlocking the Mad2 C-terminal tail is required for ligand release from Mad2, and that the 'safety belt' mechanism may prolong the lifetime of Mad2-ligand complexes. C1 European Inst Oncol, Dept Expt Oncol, Struct Biol Unit, I-20141 Milan, Italy. Pharmacia Corp, Dept Chem, Discovery Res Oncol, I-20014 Nerviano, Italy. NIH, Mol Virol Sect, Bethesda, MD 20892 USA. RP Musacchio, A (reprint author), European Inst Oncol, Dept Expt Oncol, Struct Biol Unit, Via Ripamonti 435, I-20141 Milan, Italy. RI Jeang, Kuan-Teh/A-2424-2008 NR 46 TC 182 Z9 188 U1 1 U2 12 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD MAY 15 PY 2002 VL 21 IS 10 BP 2496 EP 2506 DI 10.1093/emboj/21.10.2496 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 554TQ UT WOS:000175753600019 PM 12006501 ER PT J AU Praetorius, J Spring, KR AF Praetorius, J Spring, KR TI Specific lectins map the distribution of fibronectin and beta 1-integrin on living MDCK cells SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE confocal microscopy; laser scanning cytometry; immunocytochemistry; lectins ID CANINE KIDNEY-CELLS; BRANCHING MORPHOGENESIS; SIALIDASE TREATMENT; TUBULE FORMATION; RAT-KIDNEY; INTEGRIN; BINDING; DEPOSITION; EXPRESSION AB The expression and dynamics of bound fibronectin and the sialylated integral membrane protein, beta1-integrin, were analyzed on the apical membrane of living MDCK cells. Fibronectin was identified by its specific binding of fluorescent peanut agglutinin and sialylated,beta1-integrin by its binding of Sambucus nigra agglutinin. Confocal epifluoreseence microscopy and laser scanning cytometry determined the distribution and abundance of binding sites of the two fluorescently labeled lectins. Both fibronectin and beta1-integrin were restricted to specific regions uniformly distributed over the entire apical surface. Apical-surface fibronectin binding varied much more between cells than did the expression of beta1-integrin. Sialylated beta1-integrin colocalized >92% with membrane microplicae while fibronectin was unrelated to these surface structures. This lack of colocalization of the proteins was confirmed by double-labeling experiments. From the maturation dependence of the fibronectin-binding capacity and the differences in protein turnover times, it was evident that fibronectin did not bind to sialylated,beta1-integrin. Furthermore, desialylation of beta1-integrin uncovered additional fibronectin receptors on the apical membrane. We conclude that these lectins permit tracking of two membrane-associated glycoproteins in living cells and that fibronectin binds only to desialylated beta1-integrin on MDCK cells. C1 NHLBI, Sect Transport Physiol, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. RP Spring, KR (reprint author), NHLBI, Sect Transport Physiol, Kidney & Electrolyte Metab Lab, NIH, 10 Ctr Dr,Bldg 10,Room 6N260, Bethesda, MD 20892 USA. NR 25 TC 10 Z9 10 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD MAY 15 PY 2002 VL 276 IS 1 BP 52 EP 62 DI 10.1006/excr.2002.5516 PG 11 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 550MD UT WOS:000175506100006 PM 11978008 ER PT J AU Kasprzak, KS AF Kasprzak, KS TI Oxidative DNA and protein damage in metal-induced toxicity and carcinogenesis SO FREE RADICAL BIOLOGY AND MEDICINE LA English DT Review DE carcinogenesis; DNA; histories; metals; oxidative damage; oxygen radicals; proteins; radicals; toxicity; free radicals ID N-TERMINAL SEQUENCE; HUMAN PROTAMINE HP2; HYDROGEN-PEROXIDE; HISTONE H2A; CATALYZED OXIDATION; BINDING; NI(II); TAIL; NICKEL(II); CU(II) AB This review discusses the relevance of oxidative damage to metal-induced toxicity and carcinogenesis. Presented are important facts and mechanistic concepts on the capacity of selected transition metals, mainly Ni, but also Cu, Co, Cr, and briefly several others, to generate active oxygen species, and other reactive intermediates under physiological conditions. These metals are known to be toxic and/or carcinogenic contaminants of the occupational and general environments. Their redox activity may underlay the mechanism of mediation of oxidative damage to cell constituents. The presentation is focused on selected issues relative to genetic and epigenetic toxicity and illustrated with examples of metal-mediated oxidative damage to the principal components of chromatin, i.e., DNA, histones, and protamines. Published by Elsevier Science Inc. C1 NCI, Comparat Carcinogenesis Lab, Frederick, MD 21701 USA. RP Kasprzak, KS (reprint author), NCI, Comparat Carcinogenesis Lab, Bldg 538,Room 205E, Frederick, MD 21701 USA. NR 42 TC 142 Z9 159 U1 1 U2 17 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0891-5849 J9 FREE RADICAL BIO MED JI Free Radic. Biol. Med. PD MAY 15 PY 2002 VL 32 IS 10 BP 958 EP 967 DI 10.1016/S0891-5849(02)00809-2 PG 10 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 552GP UT WOS:000175611600002 PM 12008111 ER PT J AU Thameem, F Wolford, JK Wang, JH German, MS Bogardus, C Prochazka, M AF Thameem, F Wolford, JK Wang, JH German, MS Bogardus, C Prochazka, M TI Cloning, expression and genomic structure of human LMX1A, and variant screening in Pima Indians SO GENE LA English DT Article DE LMX1A; cloning; type 2 diabetes mellitus; single nucleotide polymorphism; candidate gene; association analysis; Pima Indians ID II DIABETES-MELLITUS; GENE-EXPRESSION; INSULIN GENE; INDEPENDENT REPLICATION; SUSCEPTIBILITY GENES; MUTATIONS; SEARCH; LOCUS; SCAN AB LIM-homeodomain containing protein LMX1A activates transcription of the insulin gene. The human LMX1A gene maps to lq22-q23, a region identified as a putative type 2 diabetes mellitus (T2DM) locus in several different populations. We analyzed LMX1A as a positional and biological candidate gene for T2DM in the Pima Indians, in whom a linkage of T2DM to 1q21-q23 has been previously reported. In the present study, we describe the cloning, expression and genomic organization of the LMX1A gene, which is composed of 11 exons spanning approximately 151 kb. In addition to a transcript encoding the predicted full-length protein of 382 amino acids, we identified two truncated cDNA forms produced via additional transcription start sites and alternative splicing. We identified seven single nucleotide polymorphisms (SNPs) throughout the LMX1A locus and determined allele frequency distributions in 150 diabetic and 150 unaffected Pimas. We did not find evidence for association of any LMX1A SNPs with T2DM and conclude that LMX1A does not contribute significantly to T2DM etiology in Pima Indians. Published by Elsevier Science B.V. C1 NIDDKD, Clin Diabet & Nutr Sect, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ 85016 USA. Univ Calif San Francisco, Hormone Res Inst, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Med, San Francisco, CA 94143 USA. RP Thameem, F (reprint author), NIDDKD, Clin Diabet & Nutr Sect, Phoenix Epidemiol & Clin Res Branch, NIH, 4212 N 16Th St, Phoenix, AZ 85016 USA. FU NIDDK NIH HHS [DK21344, R01 DK021344] NR 21 TC 10 Z9 11 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 MAY 15 PY 2002 VL 290 IS 1-2 BP 217 EP 225 AR PII S0378-1119(02)00582-6 DI 10.1016/S0378-1119(02)00582-6 PG 9 WC Genetics & Heredity SC Genetics & Heredity GA 567HN UT WOS:000176479500020 PM 12062816 ER PT J AU Qiu, HF Hu, CH Dong, JS Hinnebusch, AG AF Qiu, HF Hu, CH Dong, JS Hinnebusch, AG TI Mutations that bypass tRNA binding activate the intrinsically defective kinase domain in GCN2 SO GENES & DEVELOPMENT LA English DT Article DE eIF2 alpha kinase; GCN2; regulation; translation; tRNA ID TRANSFER-RNA-SYNTHETASES; PROTEIN-KINASE; EXPRESSION; DIMERIZATION; MECHANISM; RIBOSOMES; INTERACTS; COMPLEX; MOIETY; CELLS AB The protein kinase GCN2 is activated in amino acid-starved cells on binding of uncharged tRNA to a histidyl-tRNA synthetase (HisRS)-related domain. We isolated two point mutations in the protein kinase (PK) domain, R794G and F842L, that permit strong kinase activity in the absence of tRNA binding. These mutations also bypass the requirement for ribosome binding, dimerization, and association with the GCN1/GCN20 regulatory complex, suggesting that all of these functions facilitate tRNA binding to wild-type GCN2. While the isolated wild-type PK domain was completely inert, the mutant PK was highly active in vivo and in vitro. These results identify an inhibitory structure intrinsic to the PK domain that must be overcome on tRNA binding by interactions with a regulatory region, most likely the N terminus of the HisRS segment. As Arg 794 and Phe 842 are predicted to lie close to one another and to the active site, they may participate directly in misaligning active site residues. Autophosphorylation of the activation loop was stimulated by R794G and F842L, and the autophosphorylation sites remained critical for GCN2 function in the presence of these mutations. Our results imply a two-step activation mechanism involving distinct conformational changes in the PK domain. C1 NICHHD, Lab Gene Regulat & Dev, Bethesda, MD 20892 USA. RP Hinnebusch, AG (reprint author), NICHHD, Lab Gene Regulat & Dev, Bethesda, MD 20892 USA. NR 21 TC 29 Z9 29 U1 0 U2 2 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 MAY 15 PY 2002 VL 16 IS 10 BP 1271 EP 1280 DI 10.1101/gad.979402 PG 10 WC Cell Biology; Developmental Biology; Genetics & Heredity SC Cell Biology; Developmental Biology; Genetics & Heredity GA 555UJ UT WOS:000175811400010 PM 12023305 ER PT J AU Torrado, M Trivedi, R Zinovieva, R Karavanova, I Tomarev, SI AF Torrado, M Trivedi, R Zinovieva, R Karavanova, I Tomarev, SI TI Optimedin: a novel olfactomedin-related protein that interacts with myocilin SO HUMAN MOLECULAR GENETICS LA English DT Article ID OPEN-ANGLE GLAUCOMA; TRABECULAR MESHWORK CELLS; IN-SITU HYBRIDIZATION; MOLECULAR-CLONING; GOLGI PROTEINS; BREFELDIN-A; GENE; TIGR; GLYCOPROTEIN; LOCALIZATION AB Mutations in the MYOC gene may lead to juvenile open-angle glaucoma with high intraocular pressure, and are detected in about 4% of people with adult onset glaucoma. Most of these mutations are found in the third exon of the gene encoding the olfactomedin-like domain located at the C terminus of the protein. Another olfactomedin-related protein, known as noelin or pancortin, is involved in the generation of neural crest cells. Here we describe the identification of a novel olfactomedin-related gene, named optimedin, located on chromosome 1p21 in humans. Optimedin and noelin are both expressed in brain and retina. However, unlike noelin, rat optimedin is also highly expressed in the epithelial cells of the iris and the ciliary body in close proximity to the sites of Myoc expression. In the human eye, optimedin is expressed in the retina and the trabecular meshwork. Both optimedin and myocilin are localized in Golgi and are secreted proteins. The presence of mutant myocilin interferes with secretion of optimedin in transfected cells. Optimedin and myocilin interact with each other in vitro as judged by the GST pulldown, co-immunoprecipitation and far-western binding assays. The C-terminal olfactomedin domains are essential for interaction between optimedin and myocilin, while the N-terminal domains of both proteins are involved in the formation of protein homodimers. We suggest that optimedin may be a candidate gene for disorders involving the anterior segment of the eye and the retina. C1 NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NICHHD, Mol Neurobiol Sect, NIH, Bethesda, MD 20892 USA. RP Tomarev, SI (reprint author), NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RI Torrado, Mario/A-4889-2010 OI Torrado, Mario/0000-0001-9762-7018 NR 38 TC 69 Z9 78 U1 0 U2 0 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 MAY 15 PY 2002 VL 11 IS 11 BP 1291 EP 1301 DI 10.1093/hmg/11.11.1291 PG 11 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 556NQ UT WOS:000175856200005 PM 12019210 ER PT J AU Myerowitz, R Lawson, D Mizukami, H Mi, YD Tifft, CJ Proia, RL AF Myerowitz, R Lawson, D Mizukami, H Mi, YD Tifft, CJ Proia, RL TI Molecular pathophysiology in Tay-Sachs and Sandhoff diseases as revealed by gene expression profiling SO HUMAN MOLECULAR GENETICS LA English DT Article ID AMYOTROPHIC-LATERAL-SCLEROSIS; BONE-MARROW TRANSPLANTATION; SERIAL ANALYSIS; REACTIVE ASTROCYTES; ALZHEIMERS-DISEASE; UP-REGULATION; BRAIN; CELLS; PROTEIN; MICROGLIA AB Tay-Sachs and Sandhoff diseases are lysosomal storage disorders characterized by the absence of beta-hexosaminidase activity and the accumulation of GM2 ganglioside in neurons. In each disorder, a virtually identical course of neurodegeneration begins in infancy and leads to demise generally by 4-6 years of age. Through serial analysis of gene expression (SAGE), we determined gene expression profiles in cerebral cortex from a Tay-Sachs patient, a Sandhoff disease patient and a pediatric control. Examination of genes that showed altered expression in both patients revealed molecular details of the pathophysiology of the disorders relating to neuronal dysfunction and loss. A large fraction of the elevated genes in the patients could be attributed to activated macrophages/microglia and astrocytes, and included class II histocompatability antigens, the pro-inflammatory cytokine osteopontin, complement components, proteinases and inhibitors, galectins, osteonectin/SPARC, and prostaglandin D2 synthase. The results are consistent with a model of neurodegeneration that includes inflammation as a factor leading to the precipitous loss of neurons in individuals with these disorders. C1 NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. St Marys Coll Maryland, Dept Biol, St Marys City, MD 20686 USA. Childrens Natl Med Ctr, Dept Med Genet, Washington, DC 20010 USA. RP Proia, RL (reprint author), NIDDKD, Genet Dev & Dis Branch, NIH, Bldg 10,Room 9N-314,10 Ctr DR MSC 1821, Bethesda, MD 20892 USA. RI Proia, Richard/A-7908-2012 NR 51 TC 89 Z9 95 U1 2 U2 9 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 MAY 15 PY 2002 VL 11 IS 11 BP 1343 EP 1350 DI 10.1093/hmg/11.11.1343 PG 8 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 556NQ UT WOS:000175856200011 PM 12019216 ER PT J AU Karthikeyan, G Lewis, LK Resnick, MA AF Karthikeyan, G Lewis, LK Resnick, MA TI The mitochondrial protein frataxin prevents nuclear damage SO HUMAN MOLECULAR GENETICS LA English DT Article ID GAA TRIPLET REPEAT; SACCHAROMYCES-CEREVISIAE; FRIEDREICHS-ATAXIA; DNA-DAMAGE; IRON ACCUMULATION; YEAST FRATAXIN; OXIDATIVE STRESS; HOMOLOG; GENE; RECOMBINATION AB The mitochondrial protein frataxin helps maintain appropriate iron levels in the mitochondria of yeast and humans. A deficiency of this protein in humans causes Friedreich's ataxia, while its complete absence in yeast (Deltayfh1 mutant) results in loss of mitochondrial DNA, apparently due to radicals generated by excess iron. We found that the absence of frataxin in yeast also leads to nuclear damage, as evidenced by inducibility of a nuclear DNA damage reporter, increased chromosomal instability including recombination and mutation, and greater sensitivity to DNA-damaging agents, as well as slow growth. Addition of a human frataxin mutant did not prevent nuclear damage, although it partially complemented the Deltayfh1 mutant in preventing mitochondrial DNA loss. The effects in Deltayfh1 mutants result from reactive oxygen species (ROS), since (i) Deltayfh1 cells produce more hydrogen peroxide, (ii) the effects are alleviated by a radical scavenger and (iii) the glutathione peroxidase gene prevents an increase in mutation rates. Thus, the frataxin protein is concluded to have a protective role for the nucleus as well as the mitochondria. C1 NIEHS, Genet Mol Lab, NIH, Res Triangle Pk, NC 27709 USA. SW Texas State Univ, Dept Chem & Biochem, San Marcos, TX 78666 USA. RP NIEHS, Genet Mol Lab, NIH, POB 12233,Mail Drop D3-01,111 Alexander Dr, Res Triangle Pk, NC 27709 USA. EM Resnick@NIEHS.NIH.GOV NR 59 TC 54 Z9 55 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 EI 1460-2083 J9 HUM MOL GENET JI Hum. Mol. Genet. PD MAY 15 PY 2002 VL 11 IS 11 BP 1351 EP 1362 DI 10.1093/hmg/11.11.1351 PG 12 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 556NQ UT WOS:000175856200012 PM 12019217 ER PT J AU Wong, TY Klein, R Sharrett, AR Schmidt, MI Pankow, JS Couper, DJ Klein, BEK Hubbard, LD Duncan, BB AF Wong, TY Klein, R Sharrett, AR Schmidt, MI Pankow, JS Couper, DJ Klein, BEK Hubbard, LD Duncan, BB CA ARIC Investigators TI Retinal arteriolar narrowing and risk of diabetes mellitus in middle-aged persons SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID IMPAIRED GLUCOSE-TOLERANCE; MUSCLE BLOOD-FLOW; ATHEROSCLEROSIS RISK; MICROVASCULAR ABNORMALITIES; INSULIN-RESISTANCE; CIGARETTE-SMOKING; COMMUNITIES; HYPERTENSION; ADULTS; NIDDM AB Context Microvascular processes have been hypothesized to play a role in the pathogenesis of type 2 diabetes mellitus, but prospective clinical data regarding this hypothesis are unavailable. Objective To examine the relation of retinal arteriolar narrowing, a marker of microvascular damage from aging, hypertension, and inflammation, to incident diabetes in healthy middle-aged persons. Design, Setting, and Participants The Atherosclerosis Risk in Communities Study, an ongoing population-based, prospective cohort study in 4 US communities that began in 1987-1989. Included in this analysis were 7993 persons aged 49 to 73 years without diabetes, of whom retinal photographs were taken during the third examination (1993-1995). Main Outcome Measures Incident diabetes (defined as fasting glucose levels of greater than or equal to126 mg/dL [7.0 mmol/L], casual levels of greater than or equal to200 mg/dL [11.1 mmol/L], diabetic medications use, or physician diagnosis of diabetes at the fourth examination) by quartile of retinal arteriole-to-venule ratio (AVR). Results After a median follow-up of 3.5 years, 291 persons (3.6%) had incident diabetes. The incidence of diabetes was higher in persons with lower AVR at baseline (2.4%, 3.1%,4.0%, and 5.2%, from highest to lowest AVR quartile; P for trend<.001). After controlling for fasting glucose and insulin levels, family history of diabetes, adiposity, physical activity, blood pressure, and other factors, persons in the lowest quartile of AVR were 71 % more likely to develop diabetes than those in the highest quartile (odds ratio [OR], 1.71; 95% confidence interval [CI], 1.13-2.57; P for trend=.002). This association persisted with different diagnostic criteria (OR, 1.92; 95% CI, 1.10-3.36; P for trend=.01, using a fasting glucose level of &GE;141 mg/dL [7,8 mmol/L] as a cutoff), and was seen even in people at lower risk of diabetes, including those without a family history of diabetes, without impaired fasting glucose, and with lower measures of adiposity. Conclusions Retinal arteriolar narrowing is independently associated with risk of diabetes, supporting a microvascular role in the development of clinical diabetes. C1 Univ Wisconsin, Dept Ophthalmol, Madison, WI USA. Singapore Natl Eye Ctr, Singapore, Singapore. NHLBI, Bethesda, MD 20892 USA. Univ Fed Rio Grande Sul, Sch Med, Dept Social Med, Porto Alegre, RS, Brazil. Univ Minnesota, Div Epidemiol, Minneapolis, MN USA. Univ N Carolina, Dept Biostat, Chapel Hill, NC USA. RP Wong, TY (reprint author), Natl Univ Singapore, Dept Ophthalmol, 10 Kent Ridge Crescent, Singapore 119260, Singapore. OI Pankow, James/0000-0001-7076-483X FU NHLBI NIH HHS [N01-HC-55022, N01-HC-35125, N01-HC-55015, N01-HC-55016, N01-HC-55018, N01-HC-55019, N01-HC-55020, N1-HC-55021, NN01-HC-35126] NR 34 TC 173 Z9 178 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 MAY 15 PY 2002 VL 287 IS 19 BP 2528 EP 2533 DI 10.1001/jama.287.19.2528 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 551MV UT WOS:000175566200021 PM 12020333 ER PT J AU Tran, H Pankov, R Tran, SD Hampton, B Burgess, WH Yamada, KM AF Tran, H Pankov, R Tran, SD Hampton, B Burgess, WH Yamada, KM TI Integrin clustering induces kinectin accumulation SO JOURNAL OF CELL SCIENCE LA English DT Article DE adhesion; integrin; kinectin; kinesin-binding proteins; endoplasmic reticulum; fibronectin ID ANCHORAGE-DEPENDENT FIBROBLASTS; RECEPTOR-ASSOCIATED PROTEIN; KINESIN-BINDING PROTEIN; AMINO-ACID-SEQUENCE; CELL-ADHESION; FOCAL ADHESIONS; FIBRONECTIN RECEPTOR; EXTRACELLULAR-MATRIX; SIGNAL-TRANSDUCTION; MESSENGER-RNA AB Integrin receptors mediate the formation of adhesion complexes and play important roles in signal transduction from the extracellular matrix. Integrin-based adhesion complexes (IAC) contain proteins that link integrins to the cytoskeleton and recruit signaling molecules, including vinculin, paxillin, focal adhesion kinase, talin and alpha-actinin. In this study, we describe a similar to160 kDa protein that is markedly enriched at IAC induced by clustering integrins with fibronectin-coated beads. Protein sequence analysis reveals that this similar to160 kDa protein is kinectin. Kinectin is an integral membrane protein found in endoplasmic reticulum, and it serves as a receptor for the motor protein kinesin. Fibronectin-induced IAC sequestered over half of the total cellular content of kinectin within 20 minutes. In addition, two other ER-resident proteins, RAP [low-density lipoprotein receptor-related protein (LRP) receptor-associated protein] and calreticulin, were found to be clustered at IAC, whereas kinesin was not. Our results identify a novel class of constituents of IAC. C1 Natl Inst Dental & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. Amer Red Cross, Jerome H Holland Lab, Dept Tissue Biol, Rockville, MD 20855 USA. RP Yamada, KM (reprint author), Natl Inst Dental & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. RI Pankov, Roumen/B-3284-2014; OI Pankov, Roumen/0000-0002-3157-3659; Yamada, Kenneth/0000-0003-1512-6805 NR 55 TC 48 Z9 51 U1 0 U2 2 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD MAY 15 PY 2002 VL 115 IS 10 BP 2031 EP 2040 PG 10 WC Cell Biology SC Cell Biology GA 560YQ UT WOS:000176109300004 PM 11973345 ER PT J AU Negash, S Wang, HS Gao, C Ledee, D Zelenka, P AF Negash, S Wang, HS Gao, C Ledee, D Zelenka, P TI Cdk5 regulates cell-matrix and cell-cell adhesion in lens epithelial cells SO JOURNAL OF CELL SCIENCE LA English DT Article DE Cdk5; p35; adhesion; lens epithelial cells; N/N1003A cells ID CYCLIN-DEPENDENT KINASE-5; PROTEIN-KINASE; NEURONAL DIFFERENTIATION; NEURITE OUTGROWTH; A-CAM; ACTIVATOR; BRAIN; EXPRESSION; COMPLEX; SUBUNIT AB Cdk5 is a member of the cyclin-dependent kinase family, which is expressed predominantly in terminally differentiated neurons. Lower levels of Cdk5 are also found in a wide variety of cell types, including the lens. Although Cdk5 has been shown to play an important role in neuronal migration and neurite outgrowth, its function in non-neuronal cells is not known. Therefore, this study was undertaken to explore the role of Cdk5 in the lens. Results showed that, within the adult mouse lens, Cdk5 was localized to the cytoplasm, especially along the lateral membranes of differentiating primary fiber cells, which suggests a role in cell-cell adhesion. Staining at the tips of elongating fiber cells was also particularly strong, suggesting a role in cell-matrix adhesion. To examine the possible role of Cdk5 in lens epithelial cell adhesion, we stably transfected N/N1003A rabbit lens epithelial cells with cDNAs for Cdk5 or a dominant-negative mutation, Cdk5-T33. Attachment to a fibronectin matrix, as measured with substrate-coated cell adhesion strips, was increased by Cdk5 overexpression, while an equivalent overexpression of Cdk5-T33 had no effect. Cdk5 also increased the rate of cell attachment and spreading as measured by electric cell-substrate impedance sensing (ECIS). In addition, Cdk5 overexpression decreased cell-cell adhesion as measured by a cell aggregation assay. These findings suggest that Cdk5 plays a role in regulating both cell-matrix and cell-cell interactions in the lens. C1 NEI, NIH, Bethesda, MD 20892 USA. Yang Ming Univ, Taipei, Taiwan. RP Zelenka, P (reprint author), NEI, NIH, Bethesda, MD 20892 USA. NR 37 TC 36 Z9 37 U1 1 U2 1 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD MAY 15 PY 2002 VL 115 IS 10 BP 2109 EP 2117 PG 9 WC Cell Biology SC Cell Biology GA 560YQ UT WOS:000176109300011 PM 11973352 ER PT J AU Halet, G Tunwell, R Balla, T Swann, K Carroll, J AF Halet, G Tunwell, R Balla, T Swann, K Carroll, J TI The dynamics of plasma membrane PtdIns(4,5)P(2) at fertilization of mouse eggs SO JOURNAL OF CELL SCIENCE LA English DT Article DE phosphatidylinositol 4,5-bisphosphate; oocyte; GFP ID CORTICAL GRANULE EXOCYTOSIS; SEA-URCHIN EGGS; INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR; PLECKSTRIN HOMOLOGY DOMAINS; PHOSPHOLIPASE-C-GAMMA; CALCIUM-RELEASE; CA2+ OSCILLATIONS; SPATIOTEMPORAL DYNAMICS; INTRACELLULAR CALCIUM; MEIOTIC MATURATION AB A series of intracellular Ca(2+) oscillations are responsible for triggering egg activation and cortical granule exocytosis at fertilization in mammals. These Ca(2+) oscillations are generated by an increase in inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)], which results from the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)]. Using confocal imaging to simultaneously monitor Ca(2+) and plasma membrane Ptdlns(4,5)P(2) in single living mouse eggs we have sought to establish the relationship between the kinetics of Ptdlns(4,5)P(2) metabolism and the Ca(2+) oscillations at fertilization. We report that there is no detectable net loss of plasma membrane Ptdlns(4,5)P(2) either during the latent period or during the subsequent Ca(2+) oscillations. When phosphatidylinositol 4-kinase is inhibited with micromolar wortmannin a limited decrease in plasma membrane Ptdlns(4,5)P(2) is detected in half the eggs studied. Although we were unable to detect a widespread loss of PtdIns(4,5)P(2), we found that fertilization triggers a net increase in plasma membrane PtdIns(4,5)P(2) that is localized to the vegetal cortex. The fertilization-induced increase in PtdIns(4,5)P(2) follows the increase in Ca(2+), is blocked by Ca(2+) buffers and can be mimicked, albeit with slower kinetics, by photoreleasing Ins(1,4,5)P(3). Inhibition of Ca(2+)-dependent exocytosis of cortical granules, without interfering with Ca(2+) transients, inhibits the PtdIns(4,5)P(2) increase. The increase appears to be due to de novo synthesis since it is inhibited by micromolar wortmannin. Finally, there is no increase in Ptdlns(4,5)P(2) in immature oocytes that are not competent to extrude cortical granules. These studies suggest that fertilization does not deplete plasma membrane Ptdlns(4,5)P(2) and that one of the pathways for increasing Ptdlns(4,5)P(2) at fertilization is invoked by exocytosis of cortical granules. C1 UCL, Dept Physiol, London WC1E 6BT, England. UCL, Dept Anat & Dev Biol, London WC1E 6BT, England. NIH, Endocrinol & Reprod Res Branch, Bethesda, MD 20892 USA. RP Carroll, J (reprint author), UCL, Dept Physiol, Gower St, London WC1E 6BT, England. EM j.carroll@ucl.ac.uk RI halet, guillaume/C-7502-2009 NR 62 TC 47 Z9 47 U1 0 U2 1 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD MAY 15 PY 2002 VL 115 IS 10 BP 2139 EP 2149 PG 11 WC Cell Biology SC Cell Biology GA 560YQ UT WOS:000176109300014 PM 11973355 ER PT J AU Liao, F Shirakawa, AK Foley, JF Rabin, RL Farber, JM AF Liao, F Shirakawa, AK Foley, JF Rabin, RL Farber, JM TI Human B cells become highly responsive to macrophage-inflammatory protein-3 alpha/CC chemokine ligand-20 after cellular activation without changes in CCR6 expression or ligand binding SO JOURNAL OF IMMUNOLOGY LA English DT Article ID RECEPTOR EXPRESSION; DENDRITIC CELLS; MOLECULAR-CLONING; T-CELLS; LYMPHOID ORGANS; PROTEIN 3-ALPHA; CUTTING EDGE; LYMPHOCYTES; MIGRATION; GENE AB CCR6 is the only known receptor for the chemokine macrophage-inflammatory protein (MIP)-3alpha/CC chemokine ligand (CCL)20. We have shown previously that CCR6 is expressed on peripheral blood B cells, but CCR6 activity on these cells is low in in vitro assays. We report that MIP-3alpha/CCL20-induced calcium flux and chemotaxis can be enhanced significantly on peripheral blood and tonsillar B cells after activation by cross-linking surface Ag receptors. Of particular interest is the fact that the enhanced activity on B cells was not associated with an increase in CCR6 expression as assessed by levels of receptor mRNA, surface staining, or MIP-3alpha/CCL20 binding sites, or by a change in the affinity of the receptor for ligand. These data convincingly demonstrate that responses to a chemokine can be regulated solely by changes in the downstream pathways for signal transduction resulting from Ag receptor activation, and establish CCR6 as an efficacious receptor on human B cells. C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Farber, JM (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10,Room 11C-104,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 62 TC 60 Z9 62 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 MAY 15 PY 2002 VL 168 IS 10 BP 4871 EP 4880 PG 10 WC Immunology SC Immunology GA 550AF UT WOS:000175480100007 PM 11994436 ER PT J AU Wu, MT Hwang, ST AF Wu, MT Hwang, ST TI CXCR5-transduced bone marrow-derived dendritic cells traffic to B cell zones of lymph nodes and modify antigen-specific immune responses SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CD34(+) HEMATOPOIETIC PROGENITORS; CORD-BLOOD DIFFERENTIATE; IN-VIVO; RETROVIRAL VECTORS; T-CELLS; CHEMOKINE; FOLLICLES; LYMPHOCYTES; MOUSE; LOCALIZATION AB Skin-derived migratory dendritic cells (DC), in contrast to bone marrow-derived DC (BMDC), express CXCR5, respond to the chemokine CXC ligand 13 (CXCL13) in vitro, and are capable of migrating to B cell zones (BCZ) in lymph nodes (LN) in vivo. Herein, we analyzed the surface phenotype of skin-derived migratory DC and found that 15-35% of MHC class IIhigh cells showed high levels of expression of CXCR5 but expressed low levels of DEC205, a suggested characteristic of dermal-type DC in mice. To study the effects of CXCR5 on the trafficking dynamics of DC, we stably expressed CXCR5 in BMDC by retroviral gene transduction. CXCR5 was detected by flow cytometry on transduced cells, which responded to CXCL13 in vitro in chemotaxis assays (3-fold over nontransduced BMDC, p < 0.01). When injected into the footpads of mice, similar to40% of injected CXCR5-BMDC were observed in BCZ of draining LN. Mice were vaccinated with CXCR5- and vector-BMDC that were pulsed with keyhole limpet hemocyanin (KLH) to induce Ag-specific cellular and humoral immune responses. Mice injected with CXCR5-BMDC (vs vector-BMDC) demonstrated marginally less footpad swelling in response to intradermal injection of KLH Interestingly, significantly higher levels of KLH-specific IgG (p < 0.05) and IgM (p < 0.01) were found in the serum of mice injected with CXCR5-BMDC compared with mice immunized with vector-transduced BMDC. Thus, CXCR5 is predominantly expressed by dermal-type DC. Moreover, CXCR5 directs BMDC to BCZ of LN in vivo and modifies Ag-specific immune responses induced by BMDC vaccination. C1 NCI, Dermatol Branch, Ctr Canc Res, Bethesda, MD 20892 USA. RP Hwang, ST (reprint author), NCI, Dermatol Branch, Ctr Canc Res, Bldg 10,Room 12N246,10 Ctr Dr,MSC 1908, Bethesda, MD 20892 USA. NR 46 TC 28 Z9 30 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 MAY 15 PY 2002 VL 168 IS 10 BP 5096 EP 5102 PG 7 WC Immunology SC Immunology GA 550AF UT WOS:000175480100033 PM 11994462 ER PT J AU Saleh, A Makrigiannis, AP Hodge, DL Anderson, SK AF Saleh, A Makrigiannis, AP Hodge, DL Anderson, SK TI Identification of a novel Ly49 promoter that is active in bone marrow and fetal thymus SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NK CELL-RECEPTOR; GENE-EXPRESSION; REPERTOIRE; LY-49; ACQUISITION; ANTIGEN; H-2D(D); ADULT; MICE AB The analysis of several Ly49 genes has identified a tissue-specific promoter adjacent to the previously defined first exon. The current study reveals the presence of an additional Ly49 promoter (Pro-1) and two noncoding exons upstream of the previously defined promoter (Pro-2). DNA sequences homologous to Pro-1 are present 4-10 kb upstream of Pro-2 in all Ly49 genes examined, and Pro-1 transcripts were detected from the Ly49a, e, g, o, and v genes. Pro-1 activity can be detected in bone marrow, embryonic thymus, freshly isolated liver NK cells, and the murine LNK cell line, but it does not function in adult thymus, sorted NK-T cells, spleen NK cells, or the EL-4 T cell line, even though these cells express Ly49 proteins. Luciferase reporter assays identified a Pro-1 core promoter region that functions in the LNK cell line but not EL-4 cells. The novel promoter is not active in mature NK cells, suggesting that Pro-1 represents an early Ly49 promoter. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Ctr Canc Res, Frederick, MD 21702 USA. RP Anderson, SK (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Intramural Res Support Program, Bldg 560,Room 31-93, Frederick, MD 21702 USA. RI Anderson, Stephen/B-1727-2012 OI Anderson, Stephen/0000-0002-7856-4266 FU NCI NIH HHS [N01-CO-12400] NR 27 TC 42 Z9 42 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 MAY 15 PY 2002 VL 168 IS 10 BP 5163 EP 5169 PG 7 WC Immunology SC Immunology GA 550AF UT WOS:000175480100042 PM 11994471 ER PT J AU Ma, LA Kovacs, JA Cargnel, A Valerio, A Fantoni, G Atzori, C AF Ma, LA Kovacs, JA Cargnel, A Valerio, A Fantoni, G Atzori, C TI Mutations in the dihydropteroate synthase gene of human-derived Pneumocystis carinii isolates from Italy are infrequent but correlate with prior sulfa prophylaxis SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 7th International Workshop on Opportunistic Protists CY JUN 13-16, 2001 CL CINCINNATI, OHIO SP Soc Protozoologists, Univ Cincinnati ID RESISTANCE; PNEUMONIA AB Mutations in the human-derived Pneumocystis carinii dihydropteroate synthase (DHPS) gene have been reported with increasing frequency and have been linked to prior sulfa prophylaxis and possible emergence of sulfa resistance. This study was done to examine the prevalence and clinical significance of P. carinii DHPS mutations in Italian patients. A previously described single-strand conformation polymorphism technique was used to identify P. carinii DHPS mutations in 107 patients with acquired immunodeficiency syndrome. Overall prevalence (8%) was low compared with that in other reports. Mutations were observed in 19% (6/31) of patients exposed to sulfa prophylaxis, compared with 4% (3/76) of patients not exposed to sulfa prophylaxis (P = 0.17). No significant association was observed between the presence of DHPS mutations and mortality, CD4 cell count, or demographic factors. The study confirms the association between DHPS mutations and prior sulfa prophylaxis and shows that the prevalence of DHPS mutations in an Italian patient population is lower than that in other populations. C1 NIH, Dept Crit Care Med, Warren Grant Magnuson Clin Ctr, Bethesda, MD 20892 USA. Luigi Sacco Hosp, Dept Infect Dis, Milan, Italy. RP Ma, LA (reprint author), NIH, Dept Crit Care Med, Warren Grant Magnuson Clin Ctr, Bldg 10,Rm 7D43,10 Ctr Dr,MSC 1662, Bethesda, MD 20892 USA. NR 14 TC 46 Z9 48 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD MAY 15 PY 2002 VL 185 IS 10 BP 1530 EP 1532 DI 10.1086/340220 PG 3 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 547CE UT WOS:000175314700023 PM 11992293 ER PT J AU Rostovtseva, TK Komarov, A Bezrukov, SM Colombini, M AF Rostovtseva, TK Komarov, A Bezrukov, SM Colombini, M TI VDAC channels differentiate between natural metabolites and synthetic molecules SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE mitochondria; planar membranes; permeation; large channel; current noise; ATP ID MITOCHONDRIAL OUTER-MEMBRANE; NEUROSPORA-CRASSA; ION-CHANNEL; ADENINE-NUCLEOTIDES; INTERMEMBRANE SPACE; PERMEABILITY; PORE; TRANSPORT; ATP; COMPARTMENTATION AB VDAC provides the major permeability pathway through the mitochondrial outer membrane by forming voltage-gated channels with pore radius of 1.2-1.5 nm. We find that VDAC can select among comparably-charged molecules with a much smaller effective radius, 0.4-0.5 nm. The molecules studied were the nucleotides, ATP, UTP, NADH and synthetic anions, tetraglutamate (T-Glu) and 1-hydroxypyrene-3,6,-trisulfonate (HPTS). VDAC channels were reconstituted into planar phospholipid membranes bathed in 1.0 M NaCl (buffered to pH 8.0). The nucleotides decreased the conductance of VDAC for NaCl demonstrating that they could permeate into the channel. In contrast. T-Glu and HPTS did not change the single-channel conductance, indicating exclusion from the channel. Reversal potential measurements report near ideal selectivity of Na+ over T-Glu. The nucleotides increased single-channel noise as they penetrated into the channel, while T-Glu had no effect. HPTS increased noise, but unlike NADH, this was not voltage-dependent when HPTS was added asymmetrically, indicating no penetration into the channel. The differences in effective size and charge cannot explain the difference in permeation characteristics. Thus VDAC must select among these based on shape and charge distribution. We propose that the electrostatic environment within the channel has been evolutionarily selected to favor the passage of adenine nucleotides. 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 40 TC 47 Z9 50 U1 0 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 MAY 15 PY 2002 VL 187 IS 2 BP 147 EP 156 DI 10.1007/s00232-001-0159-1 PG 10 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 556GH UT WOS:000175840700005 PM 12029371 ER PT J AU Ohshima, T Ogawa, M Takeuchi, K Takahashi, S Kulkarni, AB Mikoshiba, K AF Ohshima, T Ogawa, M Takeuchi, K Takahashi, S Kulkarni, AB Mikoshiba, K TI Cyclin-dependent kinase 5/p35 contributes synergistically with reelin/Dab1 to the positioning of facial branchiomotor and inferior olive neurons in the developing mouse hindbrain SO JOURNAL OF NEUROSCIENCE LA English DT Article DE Cdk5; p35; reelin; disabled-1; facial branchiomotor neuron; inferior olive ID REELER-LIKE PHENOTYPE; CORTICAL-NEURONS; CEREBRAL-CORTEX; LAMINAR ORGANIZATION; MAMMALIAN BRAIN; NERVOUS-SYSTEM; OPTIC TECTUM; MUTANT MOUSE; MIGRATION; MICE AB Cyclin-dependent kinase 5 (Cdk5)/p35 is a serine/threonine kinase, and its activity is detected primarily in postmitotic neurons. Mice lacking Cdk5/p35 display migration defects of the cortical neurons in the cerebrum and cerebellum. In this study, we demonstrate that although most brainstem nuclei are found in their proper positions, the motor nucleus of the facial nerve is ectopically located and neurons of the inferior olive fail to position correctly, resulting in the lack of their characteristic structures in the hindbrain of Cdk5-/- mice. Despite the defective migration of these neurons, axonal exits of the facial nerve from brainstem and projections of the inferior cerebellar axons appear unchanged in Cdk5-/- mice. Defective neuronal migration in Cdk5-/- hindbrain was rescued by the neuron-specific expression of Cdk5 transgene. Because developmental defects of these structures have been reported in reeler and Dab1 mutant mice, we analyzed the double-null mutants of p35 and Dab1 and found more extensive ectopia of VII motor nuclei in these mice. These results indicate that Cdk5/p35 and Reelin signaling regulates the selective mode of neuronal migration in the developing mouse hindbrain. C1 RIKEN, Inst Phys & Chem Res, Dev Neurobiol Lab, Brain Sci Inst, Wako, Saitama 3510198, Japan. RIKEN, Inst Phys & Chem Res, Cell Culture Dev, Brain Sci Inst, Wako, Saitama 3510198, Japan. Natl Inst Dent & Craniofacial Res, Funct Genom Unit, NIH, Bethesda, MD 20892 USA. RP Ohshima, T (reprint author), RIKEN, Inst Phys & Chem Res, Dev Neurobiol Lab, Brain Sci Inst, 2-1 Hirosawa, Wako, Saitama 3510198, Japan. RI Mikoshiba, Katsuhiko/N-7943-2015 FU NIDCR NIH HHS [Z01DE00694-010DIR] NR 52 TC 43 Z9 48 U1 0 U2 0 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD MAY 15 PY 2002 VL 22 IS 10 BP 4036 EP 4044 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 552RY UT WOS:000175634900025 PM 12019323 ER PT J AU Akbarian, S Rios, M Liu, RJ Gold, SJ Fong, HF Zeiler, S Coppola, V Tessarollo, L Jones, KR Nestler, EJ Aghajanian, GK Jaenisch, R AF Akbarian, S Rios, M Liu, RJ Gold, SJ Fong, HF Zeiler, S Coppola, V Tessarollo, L Jones, KR Nestler, EJ Aghajanian, GK Jaenisch, R TI Brain-derived neurotrophic factor is essential for opiate-induced plasticity of noradrenergic neurons SO JOURNAL OF NEUROSCIENCE LA English DT Article DE norepinephrine; locus coeruleus; Cre recombinase; loxP; BDNF; opiates; plasticity; cAMP ID LOCUS-COERULEUS NEURONS; ELEMENT-BINDING PROTEIN; AMP RESPONSE ELEMENT; MESSENGER-RNA; TYROSINE-HYDROXYLASE; MORPHINE-WITHDRAWAL; NERVOUS-SYSTEM; IN-VITRO; TRANSCRIPTION FACTOR; TRKB RECEPTORS AB Chronic opiate exposure induces numerous neurochemical adaptations in the noradrenergic system, including upregulation of the cAMP-signaling pathway and increased expression of tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. These adaptations are thought to compensate for opiate-mediated neuronal inhibition but also contribute to physical dependence, including withdrawal after abrupt cessation of drug exposure. Little is known about molecules that regulate the noradrenergic response to opiates. Here we report that noradrenergic locus ceruleus (LC) neurons of mice with a conditional deletion of BDNF in postnatal brain respond to chronic morphine treatment with a paradoxical downregulation of cAMP-mediated excitation and lack of dynamic regulation of TH expression. This was accompanied by a threefold reduction in opiate withdrawal symptoms despite normal antinociceptive tolerance in the BDNF-deficient mice. Although expression of TrkB, the receptor for BDNF, was high in the LC, endogenous BDNF expression was absent there and in the large majority of other noradrenergic neurons. Therefore, a BDNF-signaling pathway originating from non-noradrenergic sources is essential for opiate-induced molecular adaptations of the noradrenergic system. C1 Univ Massachusetts, Sch Med, Dept Psychiat, Brudnick Neuropsychiat Res Inst, Worcester, MA 01613 USA. Whitehead Inst Biomed Res, Cambridge, MA 02142 USA. Yale Univ, Dept Psychiat, New Haven, CT 06520 USA. Univ Texas SW, Dept Psychiat, Dallas, TX 75390 USA. Univ Colorado, Dept Mol Cellular & Dev Biol, Boulder, CO 80309 USA. NCI, Frederick, MD 21702 USA. RP Akbarian, S (reprint author), Univ Massachusetts, Sch Med, Dept Psychiat, Brudnick Neuropsychiat Res Inst, 303 Belmont St, Worcester, MA 01613 USA. RI Coppola, Vincenzo/E-2917-2011 OI Coppola, Vincenzo/0000-0001-6163-1779 FU NCI NIH HHS [5-R35-CA44339]; NIDA NIH HHS [1-K08-DA00479] NR 69 TC 84 Z9 89 U1 0 U2 1 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 MAY 15 PY 2002 VL 22 IS 10 BP 4153 EP 4162 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 552RY UT WOS:000175634900035 PM 12019333 ER PT J AU Elmer, GI Pieper, JO Rubinstein, M Low, MJ Grandy, DK Wise, RA AF Elmer, GI Pieper, JO Rubinstein, M Low, MJ Grandy, DK Wise, RA TI Failure of intravenous morphine to serve as an effective instrumental reinforcer in dopamine D2 receptor knock-out mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE self-administration; morphine; dopamine; D2 receptor; mice; knock-out ID VENTRAL TEGMENTAL AREA; MU-OPIOID RECEPTORS; DEFICIENT MICE; NUCLEUS-ACCUMBENS; D-2 RECEPTOR; OPIATE REWARD; RATS; HEROIN; STRIATUM; OPERANT AB The rewarding effects of opiates are thought to be mediated through dopaminergic mechanisms in the ventral tegmental area, dopamine-independent mechanisms in the nucleus accumbens, or both. The purpose of the present study was to explore the contribution of dopamine to opiate-reinforced behavior using D2 receptor knock-out mice. Wild-type, heterozygous, and D2 knock-out mice were first trained to lever press for water reinforcement and then implanted with intravenous catheters. The ability of intravenously delivered morphine to maintain lever pressing in these mice was studied under two schedules of reinforcement: a fixed ratio 4 (FR4) schedule (saline, 0.1, 0.3, or 1.0 mg/kg, per injection) and a progressive ratio (PR) schedule (1.0 mg/kg, per injection). In the wild-type and heterozygous mice, FR4 behavior maintained by morphine injections was significantly greater than behavior maintained by vehicle injections. Response rate was inversely related to injection dose and increased significantly in the wild-type and heterozygous mice when the animals were placed on the PR schedule. In contrast, the knock-out mice did not respond more for morphine than for saline and did not respond more when increased ratios were required by the PR schedule. Thus, morphine served as a positive reinforcer in the wild-type and heterozygous mice but failed to do so in the knock-out mice. Under this range of doses and response requirements, the rewarding effects of morphine appear to depend critically on an intact D2 receptor system. C1 Univ Maryland, Sch Med, Dept Psychiat, Maryland Psychiat Res Ctr,Neurosci Program, Baltimore, MD 21228 USA. NIDA, Behav Neurosci Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Consejo Nacl Invest Cient & Tecn, RA-1428 Buenos Aires, DF, Argentina. Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Ciencias Biol, RA-1428 Buenos Aires, DF, Argentina. Oregon Hlth & Sci Univ, Vollum Inst, Portland, OR 97201 USA. Oregon Hlth & Sci Univ, Dept Physiol & Pharmacol, Portland, OR 97201 USA. RP Elmer, GI (reprint author), Univ Maryland, Sch Med, Dept Psychiat, Maryland Psychiat Res Ctr,Neurosci Program, POB 21247,Maple & Locust St, Baltimore, MD 21228 USA. RI Wise, Roy/A-6465-2012 FU NIDA NIH HHS [DA11888] NR 39 TC 55 Z9 55 U1 0 U2 1 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 MAY 15 PY 2002 VL 22 IS 10 AR RC224 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 552RY UT WOS:000175634900004 PM 12006606 ER PT J AU Feng, WY Chan, KK Covey, JM AF Feng, WY Chan, KK Covey, JM TI Electrospray LC-MS/MS quantitation, stability, and preliminary pharmacokinetics of bradykinin antagonist polypeptide B201 (NSC 710295) in the mouse SO JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS LA English DT Article DE LC-MS/MS assay; ion-trap mass spectrometry; B201 polypeptide; bradykinin antagonist ID IONIZATION MASS-SPECTROMETRY; ION-TRAP; TRIPLE QUADRUPOLE; PLASMA; URINE; MS; PEPTIDE AB B201 (NSC 710295), [SUIM-(Darg-Arg-Pro-Hyp-Gly-Igl-Ser-Digl-Oic-Arg)(2)], a third generation of bradykinin (BK) antagonist, has been found to possess high potency. We report the development of a highly sensitive electrospray LC-MS/MS assay method for the analysis of B201 in plasma for the first time, using an ion-trap mass spectrometer. Human or mouse plasma (0.2 ml) was spiked with B201 and the internal standard, substance P. The compounds were extracted with a preconditioned C-18 reversed-phase column and analyzed by LC-MSJMS. The analytes were separated on a 50 x 2 mm (i.d.) BetaBasic C8 column, using a gradient elution. The positive ion selected reaction monitor mode was used monitoring the transitions of ions at m/z 938.9(3+) --> 816.0(2+) for B201 and 674.3(2+) --> 665.7(2+) for substance P. Assay validation was performed, and the limit of quantitation (LOQ) for B201 was found to be 1 ng/ml for human plasma and 2.5 ng/ml for mouse plasma. The recovery was 78% for B201 and 88% for substance P. The assay was linear from 2.5 to 1500 ng/ml for mouse plasma monitored. Using a 0.2 ml plasma, the within-day CVs were 9.3% at 2.5 ng/ml, 6.5% at 100 ng/ml, and 3.8% at 1000 ng/ml for human plasma (n = 6). For mouse plasma, the respective within-day CVs were 17.6, 9.6, and 6.2% (n = 6). The between-day CVs for human plasma were 8.2, 10.9, and 2.4%, respectively, (n = 3) and the respective values for mouse plasma were 11.9, 8.6 and 6.5% (n = 6). Pharmacokinetics of B201 in the mouse was studied following i.v. administration at 5 ma/kg and found to conform to a two-compartment model with an initial half-life of 14 min and a terminal half-life of 44 h. Plasma B201 peak level was detected at mug/ml range and the levels were detectable for a least 24 h. Preliminary oral bioavailability was found to be about 1%. This method demonstrates that an ion trap mass spectrometer can be a powerful tool to quantify large peptides at low nanogram per mililiter with a non-isotopically labeled internal standard, (C) 2002 Elsevier Science B.V. All rights reserved. C1 Ohio State Univ, Coll Pharm, Columbus, OH 43210 USA. Ohio State Univ, Coll Med, Columbus, OH 43210 USA. Natl Canc Inst, Rockville, MD 20852 USA. RP Ohio State Univ, Coll Pharm, Columbus, OH 43210 USA. EM wyfeng2k@yahoo.com; chan.56@osu.edu; coveyj@mail.nih.gov FU NCI NIH HHS [N01-CM-07019] NR 23 TC 19 Z9 23 U1 0 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0731-7085 EI 1873-264X J9 J PHARMACEUT BIOMED JI J. Pharm. Biomed. Anal. PD MAY 15 PY 2002 VL 28 IS 3-4 BP 601 EP 612 AR PII S0731-7085(01)00697-5 DI 10.1016/S0731-7085(01)00697-5 PG 12 WC Chemistry, Analytical; Pharmacology & Pharmacy SC Chemistry; Pharmacology & Pharmacy GA 561TZ UT WOS:000176160000021 PM 12008139 ER PT J AU Wu, HW Wang, SM Brooks, BR AF Wu, HW Wang, SM Brooks, BR TI Direct observation of the folding and unfolding of a beta-hairpin in explicit water through computer simulation SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID MOLECULAR-DYNAMICS SIMULATION; AQUEOUS-SOLUTION; PEPTIDE; PROTEINS; SOLVENT; MODEL C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Wu, HW (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 50,Room 3308, Bethesda, MD 20892 USA. RI Wang, Shaomeng/E-9686-2010 NR 21 TC 16 Z9 16 U1 0 U2 0 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 MAY 15 PY 2002 VL 124 IS 19 BP 5282 EP 5283 DI 10.1021/ja0257321 PG 2 WC Chemistry, Multidisciplinary SC Chemistry GA 550UD UT WOS:000175521200015 ER PT J AU London, SJ Yuan, JM Travlos, GS Gao, YT Wilson, RE Ross, RK Yu, MC AF London, SJ Yuan, JM Travlos, GS Gao, YT Wilson, RE Ross, RK Yu, MC TI Insulin-like growth factor I, IGF-binding protein 3, and lung cancer risk in a prospective study of men in China SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID FACTOR (IGF)-I; BREAST-CANCER; CARCINOMA CELLS; PLASMA-LEVELS; EXPRESSION; MORTALITY; WOMEN; INTERVENTION; INHIBITION; PATHWAY AB Background: Insulin-like growth factor I (IGF-I) stimulates cell proliferation and inhibits apoptosis in the lung and other tissues by interacting with the IGF-I receptor. The major binding protein for IGF-I, insulin-like growth factor-binding protein 3 (IGFBP-3), modulates the effects of IGF-I but also inhibits cell growth and induces apoptosis independent of IGF-I and its receptor. In a prospective study of men in Shanghai, China, we examined the association between serum levels of IGF-I and IGFBP-3 and the subsequent risk of lung cancer. Methods: From 1986 to 1989, serum was collected from 18 244 men aged 45-64 years living in Shanghai without a history of cancer. We analyzed IGF-I and IGFBP-3 levels in serum from 230 case patients who developed incident lung cancer during follow-up and from 740 control subjects. Results: Among 230 case patients and 659 matched control subjects, increased IGF-I levels were not associated with increased risk of lung cancer. However, for subjects in the highest quartile relative to the lowest quartile of IGFBP-3, the odds ratio (OR) for lung cancer, adjusted for smoking and IGF-I, was 0.50 (95% confidence interval [CI] = 0.25 to 1.02). When the analysis was restricted to ever smokers (184 case patients and 344 matched control subjects), the OR for lung cancer in men in the highest quartile of IGFBP-3 relative to those in the lowest quartile, adjusted for smoking and IGF-I, was 0.41 (95% CI = 0.18 to 0.92). Conclusions: In this prospective study of Chinese men, higher serum levels of IGF-I did not increase the risk of lung cancer. However, subjects with higher serum levels of IGFBP-3 were at reduced risk of lung cancer. This finding is consistent with experimental data that indicate that IGFBP-3 can inhibit cellular proliferation and induce apoptosis independent of IGF-I and the IGF-I receptor. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. NIEHS, Pulm Pathobiol Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. Univ So Calif, Keck Sch Med, Norris Cotton Canc Ctr, Los Angeles, CA USA. Shanghai Canc Inst, Shanghai, Peoples R China. RP London, SJ (reprint author), NIEHS, Epidemiol Branch, POB 12233,MD A3-05, Res Triangle Pk, NC 27709 USA. OI Yuan, Jian-Min/0000-0002-4620-3108; London, Stephanie/0000-0003-4911-5290 FU NCI NIH HHS [R35 CA53890, R01 CA043092, R01 CA043092-20, R01 CA43092]; NIEHS NIH HHS [5P30ES07048, Z01 ES49017] NR 37 TC 75 Z9 82 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD MAY 15 PY 2002 VL 94 IS 10 BP 749 EP 754 PG 6 WC Oncology SC Oncology GA 552MD UT WOS:000175623400011 PM 12011225 ER PT J AU Pedeux, R Boniol, M Autier, P Dore, JF AF Pedeux, R Boniol, M Autier, P Dore, JF TI Re: DNA repair, dysplastic nevi, and sunlight sensitivity in the development of cutaneous malignant melanoma SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Letter ID CANCER C1 INSERM Unite 453, Lyon, France. Ctr Rech Publ Sante, Luxembourg, Luxembourg. RP Pedeux, R (reprint author), NCI, Human Carcinogenesis Lab, NIH, 37 Convent Dr,Bldg 37,Rm 2C26, Bethesda, MD 20892 USA. RI Boniol, Mathieu/F-9623-2011; Autier, Philippe/A-4402-2014 OI Boniol, Mathieu/0000-0003-1052-5604; Autier, Philippe/0000-0003-1538-5321 NR 6 TC 1 Z9 1 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD MAY 15 PY 2002 VL 94 IS 10 BP 772 EP 773 PG 2 WC Oncology SC Oncology GA 552MD UT WOS:000175623400016 PM 12011229 ER PT J AU Landi, MT Baccarelli, A Pesatori, AC Hedayati, M Grossman, L Tucker, MA Tarone, RE AF Landi, MT Baccarelli, A Pesatori, AC Hedayati, M Grossman, L Tucker, MA Tarone, RE TI Re: DNA repair, dysplastic nevi, and sunlight sensitivity in the development of cutaneous malignant melanoma - Response SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Letter ID SOS RESPONSE C1 NCI, Genet Epidemiol Branch, Div Canc Epidemiol & Genet, NIH, Bethesda, MD 20892 USA. Univ Milan, Epidemiol Res Ctr, Milan, Italy. Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Biochem, Baltimore, MD USA. NIH, Biostat Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Landi, MT (reprint author), NCI, Genet Epidemiol Branch, Div Canc Epidemiol & Genet, NIH, 6120 Execut Blvd,EPS 7114, Bethesda, MD 20892 USA. RI Tucker, Margaret/B-4297-2015 NR 6 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD MAY 15 PY 2002 VL 94 IS 10 BP 773 EP 774 PG 2 WC Oncology SC Oncology GA 552MD UT WOS:000175623400017 ER PT J AU Barisoni, L Kopp, JB AF Barisoni, L Kopp, JB TI Modulation of podocyte phenotype in collapsing glomerulopathies SO MICROSCOPY RESEARCH AND TECHNIQUE LA English DT Article DE HIV; detachment; FSGS ID FOCAL SEGMENTAL GLOMERULOSCLEROSIS; HIV-ASSOCIATED NEPHROPATHY; BASEMENT-MEMBRANE COMPONENTS; GLOMERULAR SLIT DIAPHRAGM; NEPHROTIC SYNDROME; TRANSGENIC MICE; NEPHRIN LOCALIZES; EPITHELIAL-CELLS; IN-VITRO; EXPRESSION AB Podocytes are well-differentiated postmitotic cells whose function is largely based on their complex cytoskeletal architecture. In diseases with proteinuria, podocytes undergo morphologic changes. Podocytes react to an injurious stimulus by a reorganization of their foot process architecture that is independent of the primary injury and the cause of the proteinuria. Collapsing glomerulopathies, including the idiopathic and secondary forms due to HIV infection, have been previously considered a part of the focal sclerosing glomerulosclerosis (FSGS) spectrum. However, in contrast to FSGS, both forms of collapsing glomerulopathy are characterized by segmental and global collapse of the glomerular basement membrane (GBM) and by characteristic ultrastructural alterations in podocytes. These alterations include loss of the actin-based cytoskeleton, a dysregulated/dedifferentiated phenotype, cellular hypertrophy, and cell proliferation. These observations raise the following questions: 1) What mechanism causes glomerular collapse and do podocytes have a role? We recently proposed that in collapsing glomerulopathies the composition of the GBM is altered and contains more immature forms of collagen IV. These observations suggest that dedifferentiated/dysregulated podocytes may participate in remodeling the GBM composition, producing fetal collagen isoforms, 2) What is the pathomechanism underlying podocyte dysregulation? Although it is still unclear which etiologic factors are responsible for the idiopathic forms of collapsing glomerulopathy, in situ hybridization studies in a transgenic mouse model of HIV-associated collapsing glomerulopathy and on renal biopsies of patients with HIV-associated collapsing glomerulopathy demonstrated the presence of the HIV-1 RNA in podocytes and tubular epithelial cells. These findings suggest a direct link between viral gene expression and the dysregulation of the podocyte phenotype. 3) Another open question is how podocytes become infected in HIV-associated collapsing glomerulopathy. HIV-1 typically uses CD4 and a co-receptor such as CCR5 or CXCR4 to enter cells. So far, there is no demonstration of the expression of these receptors in podocytes. These negative findings, however, do not exclude the possibility that in the kidney another, CD4 independent, co-receptor may be used for viral cell entry. Finally, is it important to mention that collapsing glomerulopathies have a high prevalence in black patients, suggesting a link between racial background and the virus-related podocyte injury. Published 2002 Wiley-Liss, Inc.(+) C1 NIDDK, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Dept Pathol, Baltimore, MD USA. RP Barisoni, L (reprint author), NIDDK, Kidney Dis Sect, NIH, Bld10 3N116, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 59 TC 38 Z9 44 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1059-910X J9 MICROSC RES TECHNIQ JI Microsc. Res. Tech. PD MAY 15 PY 2002 VL 57 IS 4 BP 254 EP 262 DI 10.1002/jemt.10084 PG 9 WC Anatomy & Morphology; Biology; Microscopy SC Anatomy & Morphology; Life Sciences & Biomedicine - Other Topics; Microscopy GA 555UW UT WOS:000175812500014 PM 12012394 ER PT J AU Rogozin, IB Makarova, KS Murvai, J Czabarka, E Wolf, YI Tatusov, RL Szekely, LA Koonin, EV AF Rogozin, IB Makarova, KS Murvai, J Czabarka, E Wolf, YI Tatusov, RL Szekely, LA Koonin, EV TI Connected gene neighborhoods in prokaryotic genomes SO NUCLEIC ACIDS RESEARCH LA English DT Article ID COLI RNA DEGRADOSOME; ESCHERICHIA-COLI; SELFISH OPERONS; CONTEXT; EVOLUTION; PREDICTION; CLUSTERS; PROTEIN; ORDER; CONSERVATION AB A computational method was developed for delineating connected gene neighborhoods in bacterial and archaeal genomes. These gene neighborhoods are not typically present, in their entirety, in any single genome, but are held together by overlapping, partially conserved gene arrays. The procedure was applied to comparing the orders of orthologous genes, which were extracted from the database of Clusters of Orthologous Groups of proteins (COGs), in 31 prokaryotic genomes and resulted in the identification of 188 clusters of gene arrays, which included 1001 of 2890 COGs. These clusters were projected onto actual genomes to produce extended neighborhoods including additional genes, which are adjacent to the genes from the clusters and are transcribed in the same direction, which resulted in a total of 2387 COGs being included in the neighborhoods. Most of the neighborhoods consist predominantly of genes united by a coherent functional theme, but also include a minority of genes without an obvious functional connection to the main theme. We hypothesize that although some of the latter genes might have unsuspected roles, others are maintained within gene arrays because of the advantage of expression at a level that is typical of the given neighborhood. We designate this phenomenon 'genomic hitchhiking'. The largest neighborhood includes 79 genes (COGs) and consists of overlapping, rearranged ribosomal protein superoperons; apparent genome hitchhiking is particularly typical of this neighborhood and other neighborhoods that consist of genes coding for translation machinery components. Several neighborhoods involve previously undetected connections between genes, allowing new functional predictions. Gene neighborhoods appear to evolve via complex rearrangement, with different combinations of genes from a neighborhood fixed in different lineages. C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. Univ S Carolina, Dept Math, Columbia, SC 29208 USA. RP Koonin, EV (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. NR 37 TC 106 Z9 111 U1 2 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD MAY 15 PY 2002 VL 30 IS 10 BP 2212 EP 2223 DI 10.1093/nar/30.10.2212 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551YM UT WOS:000175591600013 PM 12000841 ER PT J AU Hoover, DM Lubkowski, J AF Hoover, DM Lubkowski, J TI DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis SO NUCLEIC ACIDS RESEARCH LA English DT Article ID MOLECULAR-CLONING; CC-CHEMOKINE; ESCHERICHIA-COLI; VIRUS; DNA; POLYMERASE; EXPRESSION; INFLAMMATION; SEQUENCE; PROTEIN AB The availability of sequences of entire genomes has dramatically increased the number of protein targets, many of which will need to be overexpressed in cells other than the original source of DNA. Gene synthesis often provides a fast and economically efficient approach. The synthetic gene can be optimized for expression and constructed for easy mutational manipulation without regard to the parent genome. Yet design and construction of synthetic genes, especially those coding for large proteins, can be a slow, difficult and confusing process. We have written a computer program that automates the design of oligonucleotides for gene synthesis. Our program requires simple input information, i.e. amino acid sequence of the target protein and melting temperature (needed for the gene assembly) of synthetic oligonucleotides. The program outputs a series of oligonucleotide sequences with codons optimized for expression in an organism of choice. Those oligonucleotides are characterized by highly homogeneous melting temperatures and a minimized tendency for hairpin formation. With the help of this program and a two-step PCR method, we have successfully constructed numerous synthetic genes, ranging from 139 to 1042 bp. The approach presented here simplifies the production of proteins from a wide variety of organisms for genomics-based studies. C1 NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. RP Lubkowski, J (reprint author), NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. NR 31 TC 269 Z9 289 U1 4 U2 31 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 MAY 15 PY 2002 VL 30 IS 10 AR e43 DI 10.1093/nar/30.10.e43 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551YM UT WOS:000175591600020 PM 12000848 ER PT J AU Subramaniam, S Hirai, T Henderson, R AF Subramaniam, S Hirai, T Henderson, R TI From structure to mechanism: electron crystallographic studies of bacteriorhodopsin SO PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY OF LONDON SERIES A-MATHEMATICAL PHYSICAL AND ENGINEERING SCIENCES LA English DT Article DE electron microscopy; proton pump; image analysis; conformational change; two-dimensional crystal ID TRANSFORM INFRARED-SPECTROSCOPY; X-RAY-DIFFRACTION; ANGSTROM RESOLUTION; PROTON TRANSLOCATION; CONFORMATIONAL-CHANGES; MOLECULAR MECHANISM; MEMBRANE-PROTEIN; PURPLE MEMBRANE; PHOTOCYCLE; INTERMEDIATE AB Bacteriorhodopsin is a protein found in cell membranes of the organism H. salinarum, where it functions as an efficient light-driven proton pump. Because bacteriorhodopsin is one of the simplest ion pumps known in biology, it has been the subject of intensive investigations over the last three decades, using methods spanning the range from femtosecond spectroscopy and crystallography to biochemistry and molecular biology. Here, we focus on the structural basis for the function of this protein, with primary emphasis on the contributions of electron microscopy and crystallography towards unravelling the mechanism of vectorial proton pumping. C1 NCI, Biochem Lab, Bethesda, MD 20817 USA. MRC, Mol Biol Lab, Cambridge CB2 2QH, England. RP Subramaniam, S (reprint author), NCI, Biochem Lab, Bethesda, MD 20817 USA. RI Hirai, Teruhisa/G-2105-2015 OI Hirai, Teruhisa/0000-0002-2114-8149 NR 43 TC 32 Z9 34 U1 2 U2 10 PU ROYAL SOC LONDON PI LONDON PA 6 CARLTON HOUSE TERRACE, LONDON SW1Y 5AG, ENGLAND SN 1364-503X J9 PHILOS T ROY SOC A JI Philos. Trans. R. Soc. Lond. Ser. A-Math. Phys. Eng. Sci. PD MAY 15 PY 2002 VL 360 IS 1794 BP 859 EP 874 DI 10.1098/rsta.2001.0971 PG 16 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 551BL UT WOS:000175539600005 PM 12804283 ER PT J AU Luo, J Dunn, T Ewing, C Sauvageot, J Chen, YD Trent, J Isaacs, W AF Luo, J Dunn, T Ewing, C Sauvageot, J Chen, YD Trent, J Isaacs, W TI Gene expression signature of benign prostatic hyperplasia revealed by cDNA microarray analysis SO PROSTATE LA English DT Article DE benign prostatic hyperplasia; cDNA microarray; gene expression profiling ID FIBROBLAST-GROWTH-FACTOR; BINDING-PROTEINS; IGF-I; EPITHELIAL-CELLS; FACTOR-BETA; CANCER; TISSUE; ALPHA(2)-MACROGLOBULIN; CARCINOMA; ANDROGENS AB BACKGROUND. Despite the high prevalence of benign prostatic hyperplasia (BPH) in the aging male, little is known regarding the etiology of this disease. A better understanding of the molecular etiology of BPH would be facilitated by a comprehensive analysis of gene expression patterns that are characteristic of benign growth in the prostate gland. Since genes differentially expressed between BPH and normal prostate tissues are likely to reflect underlying pathogenic mechanisms involved in the development of BPH, we performed comparative gene expression analysis using cDNA microarray technology to identify candidate genes associated with BPH. METHODS. Total RNA was extracted from a set of 9 BPH specimens from men with extensive hyperplasia and a set of 12 histologically normal prostate tissues excised from radical prostatectomy specimens. Each of these 21 RNA samples was labeled with Cy3 in a reverse transcription reaction and cohybridized with a Cy5 labeled common reference sample to a cDNA microarray containing 6,560 human genes. Normalized fluorescent intensity ratios from each hybridization experiment were extracted to represent the relative mRNA abundance for each gene in each sample. Weighted gene and random permutation analyses were performed to generate a subset of genes with statistically significant differences in expression between BPH and normal prostate tissues. Semi-quantitative PCR analysis was performed to validate differential expression. RESULTS. A subset of 76 genes involved in a wide range of cellular functions was identified to be differentially expressed between BPH and normal prostate tissues. Semi-quantitative PCR,was performed on 10 genes and 8 were validated. Genes consistently upregulated in BPH when compared to normal prostate tissues included: a restricted set of growth factors and their binding proteins (e.g. IGF-1 and -2, TGF-beta3, BMP5, latent TGF-beta binding protein 1 and -2); hydrolases, proteases, and protease inhibitors (e.g. neuropathy target esterase, MMP2, alpha-2-macroglobulin); stress response enzymes (e.g. COX2, GSTM5); and extracellular matrix molecules (e.g. laminin alpha 4 and beta 1, chondroitin sulfate proteoglycan 2, lumican). Genes consistently expressing less mRNA in BPH than in normal prostate tissues were less commonly observed and included the transcription factor KLF4, thrombospondin 4, nitric oxide synthase 2A, transglutaminase 3, and gastrin releasing peptide. CONCLUSIONS. We identified a diverse set of genes that are potentially related to benign prostatic hyperplasia, including genes both previously implicated in BPH pathogenesis as well as others not previously linked to this disease. Further targeted validation and investigations of these genes at the DNA, mRNA, and protein levels are warranted to determine the clinical relevance and possible therapeutic utility of these genes. (C) 2002 Wiley-Liss, Inc. C1 Johns Hopkins Med Inst, James Buchanan Brady Urol Inst, Dept Urol, Baltimore, MD 21287 USA. NHGRI, Canc Genet Branch, Bethesda, MD 20892 USA. RP Isaacs, W (reprint author), Johns Hopkins Med Inst, James Buchanan Brady Urol Inst, Dept Urol, 115 Marburg,600 N Wolfe St, Baltimore, MD 21287 USA. FU NIDDK NIH HHS [DK 52675] NR 51 TC 83 Z9 89 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0270-4137 J9 PROSTATE JI Prostate PD MAY 15 PY 2002 VL 51 IS 3 BP 189 EP 200 DI 10.1002/pros.10087 PG 12 WC Endocrinology & Metabolism; Urology & Nephrology SC Endocrinology & Metabolism; Urology & Nephrology GA 547XC UT WOS:000175357200004 PM 11967953 ER PT J AU London, RE Gabel, SA AF London, RE Gabel, SA TI Formation of a trypsin-borate-4-aminobutanol ternary complex SO BIOCHEMISTRY LA English DT Article ID GAMMA-GLUTAMYL-TRANSPEPTIDASE; NUCLEAR-MAGNETIC-RESONANCE; CONTAINING THROMBIN INHIBITORS; ACTIVE-CENTER HISTIDINE; SERINE PROTEASES; ALPHA-CHYMOTRYPSIN; PEPTIDEBORONIC ACIDS; H-1-NMR SPECTROSCOPY; NMR-SPECTROSCOPY; DIETARY BORON AB The formation of ternary complexes involving serine proteases, borate, and an alcohol has important implications for understanding the physiological actions of borate and for the development of tight binding inhibitors for this class of enzymes. Recent studies of a related enzyme, gamma-glutamyl transpeptidase, which is subject to inhibition by a labile serine/borate mixture, have demonstrated that construction of a nonlabile boronate analogue results in an inhibitor with nearly 10(5)-fold greater potency. To evaluate the generalization of this biochemistry to serine proteases, we have observed the ternary conplex formed from 4-aminobutanol, borate, and trypsin. A combination of B-11 and H-1 NMR and spectrophotometric assays using acetylarginine p-nitroanilide (Ac-Arg-pNA) as the chromogenic substrate all indicate a cooperative binding interaction in which the borate is esterified by the oxygen atoms of the 4-aminobutanol and trypsin residue Ser(195). Two downfield-shifted proton resonances at 15.5 and 16.6 ppm are proposed to arise from the labile imidazolium protons on His(57), indicating a salt bridge interaction with the negatively charged borate. A cooperativity parameter alpha of 0.2 is derived from the assays. These results provide the first direct evidence for formation of a ternary complex involving a serine protease, borate, and an alcohol, and suggest that this represents a general approach for the development of tight binding ligands. C1 NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. RP London, RE (reprint author), NIEHS, Struct Biol Lab, MR-01,111 Alexander Dr,POB 12233, Res Triangle Pk, NC 27709 USA. NR 43 TC 10 Z9 10 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD MAY 14 PY 2002 VL 41 IS 19 BP 5963 EP 5967 DI 10.1021/bi025583z PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551ER UT WOS:000175547000004 PM 11993990 ER PT J AU Barrientos, LG Gawrisch, K Cheng, N Steven, AC Gronenborn, AM AF Barrientos, LG Gawrisch, K Cheng, N Steven, AC Gronenborn, AM TI Structural characterization of the dilute aqueous surfactant solution of cetylpyridinium bromide/hexanol/sodium bromide SO LANGMUIR LA English DT Article ID RESIDUAL DIPOLAR COUPLINGS; LIQUID-CRYSTALLINE MEDIUM; BIOLOGICAL MACROMOLECULES; MAGNETIC-RESONANCE; INDUCED ALIGNMENT; TERNARY-SYSTEMS; NMR-SPECTRA; PROTEINS; PHASE; RESOLUTION AB The liquid crystalline alignment medium based on a quasi-ternary surfactant system comprising cetylpyridinium bromide (CPBr)/hexanol/sodium bromide [6.5% CPBr/hexanol (1/1.33, w/w), 30 mM NaBr in 10 mM glycine/HBr buffer, pH 3.0] was characterized. We assessed the morphology of the amphiphilic aggregates by electron microscopy. The images show a network of elongated wormlike structures that are 5.5 nm wide ranging up to 1 mum in length. In the bulk medium, they are closely packed with an average center-to-center spacing of similar to5 nm as measured by negative staining and similar to10 nm as determined directly by cryo-electron microscopy. Solid-state H-2 and P-31 NMR spectroscopies were used to determine the orientational order parameters of the aromatic ring and the aliphatic chain of the surfactant as well as the alcohol. Order parameter analysis revealed that the pyridinium ring is tilted with respect to the CPBr molecular symmetry axis. The cetyl chain has the typical order profile of hydrocarbon chains in liquid-crystalline phases: an order plateau for methylene segments near the pyridinium ring and rapidly decreasing order for the second half of the chain toward the terminal methyl group. Hexanol shows a similar order profile. However, its order is much lower than the order of the CPBr chains. To demonstrate the efficacy of this alignment medium for structure determination by NMR, we measured a large set of residual dipolar couplings for the potent HIV-1 inactivating protein, cyanovirin-N, determined the orientation of the alignment tensor, and verified the fold of this molecule at low pH. C1 NIH, NIDDK, Chem Phys Lab, Bethesda, MD 20892 USA. NIH, NIAAA, Lab Membrane Biochem & Biophys, Rockville, MD 20852 USA. NIH, NIAMS, Struct Biol Res Lab, Bethesda, MD 20892 USA. RP Gronenborn, AM (reprint author), NIH, NIDDK, Chem Phys Lab, Bethesda, MD 20892 USA. NR 39 TC 10 Z9 10 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0743-7463 J9 LANGMUIR JI Langmuir PD MAY 14 PY 2002 VL 18 IS 10 BP 3773 EP 3779 AR UNSP LA011448W DI 10.1021/la011448w PG 7 WC Chemistry, Multidisciplinary; Chemistry, Physical; Materials Science, Multidisciplinary SC Chemistry; Materials Science GA 550JR UT WOS:000175500300004 ER PT J AU Senkevich, TG White, CL Koonin, EV Moss, B AF Senkevich, TG White, CL Koonin, EV Moss, B TI Complete pathway for protein disulfide bond formation encoded by poxviruses SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID VACCINIA VIRUS L1R; LINKED SULFHYDRYL OXIDASE; ENDOPLASMIC-RETICULUM; G4L GENE; FAMILY; MORPHOGENESIS; GLUTAREDOXIN; CYSTEINES; DEFINES; MEMBER AB We show that three cytoplasmic thiol oxidoreductases encoded by vaccinia virus comprise a complete pathway for formation of disulfide bonds in intracellular virion membrane proteins. The pathway was defined by analyzing conditional lethal mutants and effects of cysteine to serine substitutions and by trapping disulfide-bonded heterodimer intermediates for each consecutive step. The upstream component, E10R, belongs to the ERV1/ALR family of FAD-containing sulfhydryl oxidases that use oxygen as the electron acceptor. The second component, A2.5L, is a small alpha-helical protein with a CxxxC motif that forms a stable disulfide-linked heterodimer with E10R and a transient disulfide-linked complex with the third component, G4L. The latter is a thioredoxin-like protein that directly oxidizes thiols of L1R, a structural component of the virion membrane with three stable disulfide bonds, and of the related protein F9L. These five proteins are conserved in all pox-viruses, suggesting that-the pathway is an ancestral mechanism for direct thiol-disulfide interchanges between proteins even in an unfavorable reducing environment. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. NIH, Natl Ctr Biotechnol Informat, Bethesda, MD 20892 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 25 TC 133 Z9 141 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 MAY 14 PY 2002 VL 99 IS 10 BP 6667 EP 6672 DI 10.1073/pnas.062163799 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300029 PM 11983854 ER PT J AU Mihara, H Kato, S Lacourciere, GM Stadtman, TC Kennedy, RAJD Kurihara, T Tokumoto, U Takahashi, Y Esaki, N AF Mihara, H Kato, S Lacourciere, GM Stadtman, TC Kennedy, RAJD Kurihara, T Tokumoto, U Takahashi, Y Esaki, N TI The iscS gene is essential for the biosynthesis of 2-selenouridine in tRNA and the selenocysteine-containing formate dehydrogenase H SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IN-VITRO BIOSYNTHESIS; COLI TRANSFER-RNA; NIFS-LIKE PROTEIN; ESCHERICHIA-COLI; CYSTEINE DESULFURASE; SELENIUM METABOLISM; LYASE; 4-THIOURIDINE; PURIFICATION; ENZYME AB Three NifS-like proteins, IscS, CSD, and CsdB, from Escherichia coli catalyze the removal of sulfur and selenium from L-Cysteine and L-selenocysteine, respectively, to form L-alanine. These enzymes are proposed to function as sulfur-delivery proteins for iron-sulfur cluster, thiamin, 4-thiouridine, biotin, and molybdopterin. Recently, it was reported that selenium mobilized from free selenocysteine is incorporated specifically into a selenoprotein and tRNA in vivo, supporting the involvement of the NifS-like proteins in selenium metabolism. We here report evidence that a strain lacking IscS is incapable of synthesizing 5-methylaminomethyl-2-selenouridine and its precursor 5-methylaminomethyl-2-thiouridine (mnm(5)s(2)U) in tRNA, suggesting that the sulfur atom released from L-cysteine by the action of IscS is incorporated into mnm(5)s(2)U. in contrast, neither CSD nor CsdB was essential for production of mnm(5)s(2)U and 5-methylaminomethyl-2-selenouridine. The lack of IscS also caused a significant loss of the selenium-containing polypeptide of formate dehydrogenase H. Together, these results suggest a dual function of IscS in sulfur and selenium metabolism. C1 Kyoto Univ, Inst Chem Res, Kyoto 6110011, Japan. NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. Osaka Univ, Grad Sch Sci, Dept Biol, Toyonaka, Osaka 5600043, Japan. RP Esaki, N (reprint author), Kyoto Univ, Inst Chem Res, Kyoto 6110011, Japan. RI Takahashi, Yasuhiro /F-1119-2011; Mihara, Hisaaki/F-2172-2010 OI Mihara, Hisaaki/0000-0002-5153-4403 NR 40 TC 43 Z9 48 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 MAY 14 PY 2002 VL 99 IS 10 BP 6679 EP 6683 DI 10.1073/pnas.102176099 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300031 PM 11997471 ER PT J AU Loukinov, DI Pugacheva, E Vatolin, S Pack, SD Moon, H Chernukhin, I Mannan, P Larsson, E Kanduri, C Vostrov, AA Cui, H Niemitz, EL Rasko, JEJ Docquier, FM Kistler, M Breen, JJ Zhuang, ZP Quitschke, WW Renkawitz, R Klenova, EM Feinberg, AP Ohlsson, R Morse, HC Lobanenkov, VV AF Loukinov, DI Pugacheva, E Vatolin, S Pack, SD Moon, H Chernukhin, I Mannan, P Larsson, E Kanduri, C Vostrov, AA Cui, H Niemitz, EL Rasko, JEJ Docquier, FM Kistler, M Breen, JJ Zhuang, ZP Quitschke, WW Renkawitz, R Klenova, EM Feinberg, AP Ohlsson, R Morse, HC Lobanenkov, VV TI BORIS, a novel male germ-line-specific protein associated with epigenetic reprogramming events, shares the same 11-zinc-finger domain with CTCF, the insulator protein involved in reading imprinting marks in the soma SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID C-MYC GENE; TRANSCRIPTION FACTOR; DNA-SEQUENCE; BINDING; METHYLATION; DELETIONS; PROMOTER; REGION; LOCUS; EXPRESSION AB CTCF, a conserved, ubiquitous, and highly versatile 11-zinc-finger factor involved in various aspects of gene regulation, forms methylation-sensitive insulators that regulate X chromosome inactivation and expression of imprinted genes. We document here the existence of a paralogous gene with the same exons encoding the 11-zinc-finger domain as mammalian CTCF genes and thus the same DNA-binding potential, but with distinct amino and carboxy termini. We named this gene BORIS for Brother of the Regulator of Imprinted Sites. BORIS is present only in the testis, and expressed in a mutually exclusive manner with CTCF during male germ cell development. We show here that erasure of methylation marks during male germ-line development is associated with dramatic up-regulation of BORIS and down-regulation of CTCF expression. Because BORIS bears the same DNA-binding domain that CTCF employs for recognition of methylation marks in soma, BORIS is a candidate protein for the elusive epigenetic reprogramming factor acting in the male germ line. C1 NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. NINCDS, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. Univ Essex, Dept Biol Sci, Colchester CQ4 3SQ, Essex, England. Uppsala Univ, Dept Genet & Pathol, S-75236 Uppsala, Sweden. Uppsala Univ, Dept Genet & Dev, S-75236 Uppsala, Sweden. SUNY Stony Brook, Dept Psychiat & Behav Sci, Stony Brook, NY 11794 USA. Johns Hopkins Univ, Sch Med, Inst Med Genet, Baltimore, MD 21205 USA. Centenary Inst, Gene Therapy Res Unit, Newtown, NSW 2042, Australia. Sydney Canc Ctr, Newtown, NSW 2042, Australia. Univ S Carolina, Dept Chem & Biochem, Columbia, SC 29208 USA. Univ S Carolina, Sch Med, Columbia, SC 29208 USA. Univ Giessen, Genet Inst, D-35392 Giessen, Germany. RP Lobanenkov, VV (reprint author), NIAID, Immunopathol Lab, NIH, Bldg 7,Room 303,7 Ctr Dr,MSC 0760, Bethesda, MD 20892 USA. RI Cui, Hengmi/A-2598-2008; Rasko, John/F-5754-2013; Pack, Svetlana/C-2020-2014; OI Lobanenkov, Victor/0000-0001-6665-3635; Morse, Herbert/0000-0002-9331-3705 FU NCI NIH HHS [R01 CA65145, R37 CA054358, R01 CA065145, R37 CA054358-12, R37 CA54358]; NICHD NIH HHS [HD-10793, R01 HD010793]; NINDS NIH HHS [NS30994, R01 NS030994]; Worldwide Cancer Research [99-0514] NR 31 TC 213 Z9 224 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 MAY 14 PY 2002 VL 99 IS 10 BP 6806 EP 6811 DI 10.1073/pnas.092123699 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300053 PM 12011441 ER PT J AU Sobol, RW Watson, DE Nakamura, J Yakes, FM Hou, E Horton, JK Ladapo, J Van Houten, B Swenberg, JA Tindall, KR Samson, LD Wilson, SH AF Sobol, RW Watson, DE Nakamura, J Yakes, FM Hou, E Horton, JK Ladapo, J Van Houten, B Swenberg, JA Tindall, KR Samson, LD Wilson, SH TI Mutations associated with base excision repair deficiency and methylation-induced genotoxic stress SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DNA-POLYMERASE-BETA; CHAIN-REACTION ASSAY; CANCER CELL-LINES; INDUCED CYTOTOXICITY; ALKYLATING-AGENTS; OXIDATIVE STRESS; PHOSPHATE LYASE; GENOMIC DNA; BONE-MARROW; DAMAGE AB The long-term effect of exposure to DNA alkylating agents is entwined with the cell's genetic capacity for DNA repair and appropriate DNA damage responses. A unique combination of environmental exposure and deficiency in these responses can lead to genomic instability; this "gene-environment interaction" paradigm is a theme for research on chronic disease etiology. In the present study, we used mouse embryonic fibroblasts with a gene deletion in the base excision repair (BER) enzymes DNA P-polymerase (beta-pol) and alkyladenine DNA glycosylase (AAG), along with exposure to methyl methanesulfonate (MMS) to study mutagenesis as a function of a particular gene-environment interaction. The beta-pol null cells, defective in BER, exhibit a modest increase in spontaneous mutagenesis compared with wild-type cells. MMS exposure increases mutant frequency in beta-pol null cells, but not in isogenic wild-type cells; UV light exposure or N-methyl-N'-nitro-N-nitrosoguanidine exposure increases mutant frequency similarly In both cell lines. The MMS-induced increase in mutant frequency in beta-pol null cells appears to be caused by DNA lesions that are AAG substrates, because overexpression of AAG in beta-pol null cells eliminates the effect. In contrast, beta-pol/AAG double null cells are slightly more mutable than the beta-pol null cells after MMS exposure. These results illustrate that BER plays a role in protecting mouse embryonic fibroblast cells against methylation-incluced mutations and characterize the effect of a particular combination of BER gene defect and environmental exposure. C1 NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Environm Carcinogenesis & Mutagenesis, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Environm Sci & Engn, Chapel Hill, NC 27599 USA. Univ Texas, Sealy Ctr Mol Sci, Med Branch, Galveston, TX 77555 USA. MIT, Biol Engn Div, Cambridge, MA 02139 USA. RP Wilson, SH (reprint author), NIEHS, Struct Biol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. RI Sobol, Robert/E-4125-2013; OI Sobol, Robert/0000-0001-7385-3563; Ladapo, Joseph/0000-0002-8518-4800 FU NCI NIH HHS [R01 CA055042, R01 CA075576, R01 CA55042, R01 CA75576] NR 47 TC 55 Z9 57 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 MAY 14 PY 2002 VL 99 IS 10 BP 6860 EP 6865 DI 10.1073/pnas.092662499 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300063 PM 11983862 ER PT J AU Recillas-Targa, F Pikaart, MJ Burgess-Beusse, B Bell, AC Litt, MD West, AG Gaszner, M Felsenfeld, G AF Recillas-Targa, F Pikaart, MJ Burgess-Beusse, B Bell, AC Litt, MD West, AG Gaszner, M Felsenfeld, G TI Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE CTCF; chromatin domain; boundaries ID IMPRINTED EXPRESSION; CHROMATIN INSULATOR; GENE-EXPRESSION; PROTEIN CTCF; METHYLATION; LOCUS; ELEMENT; CELLS; PROBABILITY; ASSOCIATION AB The 1.2-kb DNA sequence element (5'HS4) at the 5' end of the chicken beta-globin locus has the two defining properties of an insulator: it prevents an "external" enhancer from acting on a promoter when placed between them ("enhancer blocking") and acts as a barrier to chromosomal position effect (CPE) when it surrounds a stably integrated reporter. We previously reported that a single CTCF-bincling site in 5'HS4 is necessary and sufficient for enhancer blocking. We show here that a 250-bp "core" element from within 5'HS4 is sufficient to confer protection against silencing of transgenes caused by CPE. Further dissection of the core reveals that 5'HS4 is a compound element in which it is possible to separate enhancer blocking and barrier activities. We demonstrate that full protection against CPE is conferred by mutant 5'HS4 sequences from which the CTCF-binding site has been deleted. In contrast, mutations of four other protein binding sites within 5'HS4 result in varying reductions in the ability to protect against CPE. We find that binding sites for CTCF are neither necessary nor sufficient for protection against CPE. Comparison of the properties of 5'HS4 with those of other CTCF-bincling enhancer-blocking elements suggests that CPE protection is associated with maintenance of a high level of histone acetylation near the insulator, conferred by insulator binding-proteins other than CTCF. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Felsenfeld, G (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. OI West, Adam/0000-0003-3502-7804 NR 31 TC 269 Z9 282 U1 0 U2 6 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD MAY 14 PY 2002 VL 99 IS 10 BP 6883 EP 6888 DI 10.1073/pnas.102179399 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300067 PM 12011446 ER PT J AU Kobayashi, SD Voyich, JM Buhl, CL Stahl, RM DeLeo, FR AF Kobayashi, SD Voyich, JM Buhl, CL Stahl, RM DeLeo, FR TI Global changes in gene expression by human polymorphonuclear leukocytes during receptor-mediated phagocytosis: Cell fate is regulated at the level of gene expression SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID CHRONIC GRANULOMATOUS-DISEASE; TYROSINE PHOSPHORYLATION; NEUTROPHIL APOPTOSIS; NADPH OXIDASE; ACTIVATION; RELEASE; GLYCOSYLPHOSPHATIDYLINOSITOL; STREPTOCOCCUS; REQUIREMENT; MACROPHAGES AB Phagocytes are a critical component of the innate immune response in humans and eliminate invading microorganisms through a process known as phagocytosis. Two distinct receptor-linked phagocytic pathways, one with Ab receptors (FcRs; FcR, Fc receptor) and the other complement receptors (CRs), mediate binding and ingestion of pathogens by human polymorphonuclear leukocytes (PMNs). Although progress has been made toward defining complex signal transduction processes that underlie phagocytosis in each pathway, very little is known about gene regulation during or after phagocytosis. Therefore, we used human oligonucleotide microarrays to identify changes in expression of 12,561 genes accompanying FcR- and CR-mediated phagocytosis. Eighty-four percent of 279 differentially expressed genes were induced or repressed 90 min after ingestion of Ab- and/or complement-opsonized particles. Unexpectedly, more than 30 of these genes encoded proteins involved in at least three distinct apoptotic pathways. Ninety-four differentiaily expressed cell fate-related genes were identified between 180 and 360 min after phagocytosis and most were induced or repressed by PMNs activated through both receptors simultaneously. By using flow cytometry, we found that FcR- and CR-mediated phagocytosis each promoted programmed cell death in human PMNs; however, phagocytosis mediated by the combination of FcRs and CRs induced apoptosis earlier than that by either receptor alone. Our results reveal distinct patterns of receptor-mediated gene expression that define complex inducible apoptotic pathways in activated PMNs. Most significantly, we discovered that programmed cell death is regulated at the level of gene expression. Thus, we hypothesize that gene regulation in PMNs facilitates resolution of inflammatory responses. C1 NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, Hamilton, MT 59840 USA. RP DeLeo, FR (reprint author), NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, Hamilton, MT 59840 USA. OI DeLeo, Frank/0000-0003-3150-2516 NR 30 TC 148 Z9 155 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 MAY 14 PY 2002 VL 99 IS 10 BP 6901 EP 6906 DI 10.1073/pnas.092148299 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300070 PM 11983860 ER PT J AU Moulard, M Phogat, SK Shu, Y Labrijn, AF Xiao, XD Binley, JM Zhang, MY Sidorov, IA Broder, CC Robinson, J Parren, PWHI Burton, DR Dimitrov, DS AF Moulard, M Phogat, SK Shu, Y Labrijn, AF Xiao, XD Binley, JM Zhang, MY Sidorov, IA Broder, CC Robinson, J Parren, PWHI Burton, DR Dimitrov, DS TI Broadly cross-reactive HIV-1-neutralizing human monoclonal Fab selected for binding to gp120-CD4-CCR5 complexes SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE AIDS; antibody; vaccines; inhibitors ID IMMUNODEFICIENCY-VIRUS TYPE-1; INTERMOLECULAR DISULFIDE BOND; GP120 ENVELOPE GLYCOPROTEIN; VIRAL MEMBRANE-FUSION; CONFORMATIONAL-CHANGES; SOLUBLE CD4; INFLUENZA HEMAGGLUTININ; GP41 SUBUNITS; CELL FUSION; HIV-1 AB HIV-1 entry into cells involves formation of a complex between gp120 of the viral envelope glycoprotein (Env), a receptor (CD4), and a coreceptor, typically CCR5. Here we provide evidence that purified gp120(JR-FL)-CD4-CCR5 complexes exhibit an epitope recognized by a Fab (X5) obtained by selection of a phage display library from a seropositive donor with a relatively high broadly neutralizing serum antibody titer against an immobilized form of the trimolecular complex. X5 bound with high (W) affinity to a variety of Envs, including primary isolates from different clades and Envs with deleted variable loops (V1, -2, -3), Its binding was significantly increased by CD4 and slightly enhanced by CCR5. X5 inhibited infection of peripheral blood mononuclear cells by a selection of representative HIV-1 primary isolates from clades A, B, C, D, E, F, and G with an efficiency comparable to that of the broadly neutralizing antibody IgG1 b12. Furthermore, X5 inhibited cell fusion mediated by Envs from R5, X4, and R5X4 viruses. Of the five broadly cross-reactive HIV-1-neutralizing human monoclonal antibodies known to date, X5 is the only one that exhibits increased binding to gp120 complexed with receptors. These findings suggest that X5 could possibly be used as entry inhibitor alone or in combination with other antiretroviral drugs for the treatment of HIV-1-infected individuals, provide evidence for the existence of conserved receptor-inducible gp120 epitopes that can serve as targets for potent broadly cross-reactive neutralizing antibodies in HIV-1-infected patients, and have important conceptual and practical implications for the development of vaccines and inhibitors. C1 NCI, Lab Expt & Computat Biol, NIH, Frederick, MD 21702 USA. Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA. Uniformed Serv Univ Hlth Sci, Dept Microbiol & Immunol, Bethesda, MD 20814 USA. Louisiana State Univ, Med Ctr, Dept Pediat, New Orleans, LA 70112 USA. Scripps Res Inst, Dept Biol Mol, La Jolla, CA 92037 USA. RP Dimitrov, DS (reprint author), NCI, Lab Expt & Computat Biol, NIH, Bldg 469,Room 246,POB B,Miller Dr, Frederick, MD 21702 USA. OI Sidorov, Igor/0000-0001-6519-4983; Parren, Paul/0000-0002-4365-3859 FU NIAID NIH HHS [R37 AI033292, AI24030, AI33292, AI42599, R01 AI024030, R01 AI033292] NR 48 TC 177 Z9 187 U1 1 U2 12 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 MAY 14 PY 2002 VL 99 IS 10 BP 6913 EP 6918 DI 10.1073/pnas.102562599 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300072 PM 11997472 ER PT J AU Desai, KV Xiao, N Wang, W Gangi, L Greene, J Powell, JI Dickson, R Furth, P Hunter, K Kucherlapati, R Simon, R Liu, ET Green, JE AF Desai, KV Xiao, N Wang, W Gangi, L Greene, J Powell, JI Dickson, R Furth, P Hunter, K Kucherlapati, R Simon, R Liu, ET Green, JE TI Initiating oncogenic event determines gene-expression patterns of human breast cancer models SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE cDNA microarray; mammary cancer; oncogenes; gene-expression profiles ID LARGE T-ANTIGEN; TRANSGENIC MICE; ESTROGEN-RECEPTOR; CDNA MICROARRAY; MAMMARY-TUMORS; NEU ONCOGENE; FUSION GENE; IN-VITRO; PATHWAY; GROWTH AB Molecular expression profiling of tumors initiated by transgenic overexpression of c-myc, c-neu, c-ha-ras, polyoma middle T antigen (PyMT) or simian virus 40 T/t antigen (T-ag) targeted to the mouse mammary gland have identified both common and oncogene-specific events associated with tumor formation and progression. The tumors shared great similarities in their gene-expression profiles as compared with the normal mammary gland with an induction of cell-cycle regulators, metabolic regulators, zinc finger proteins, and protein tyrosine phosphatases, along with the suppression of some protein tyrosine kinases. Selection and hierarchical clustering of the most variant genes, however, resulted in separating the mouse models into three groups with distinct oncogene-specific patterns of gene expression. Such an identification of targets specified by particular oncogenes may facilitate development of lesion-specific therapeutics and preclinical testing. Moreover, similarities in gene expression between human breast cancers and the mouse models have been identified, thus providing an important component for the validation of transgenic mammary cancer models. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. NCI, Ctr Adv Technol, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Microarray Core Facil, Frederick, MD 21702 USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Washington, DC 20007 USA. NIH, Lab Populat Genet, Bethesda, MD 20892 USA. Harvard Partners Ctr Genet & Genomics, Boston, MA 02115 USA. RP Green, JE (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, 41 Lib Dr,Room C619, Bethesda, MD 20892 USA. RI Liu, Edison/C-4141-2008 FU NCI NIH HHS [R01 CA089041, R01 CA089041-05] NR 41 TC 150 Z9 153 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 MAY 14 PY 2002 VL 99 IS 10 BP 6967 EP 6972 DI 10.1073/pnas.102172399 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300081 PM 12011455 ER PT J AU Bera, TK Iavarone, C Kumar, V Lee, S Lee, B Pastan, I AF Bera, TK Iavarone, C Kumar, V Lee, S Lee, B Pastan, I TI MRP9, an unusual truncated member of the ABC transporter superfamily, is highly expressed in breast cancer SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID PROSTATE; GENES; DATABASE; CELLS; PROTEINS; DISCOVERY; HOMOLOGY; COMPLEX; FAMILY AB Combining a computer-based screening strategy and functional genomics, we previously identified MRP9 (ABCC12), a member of the ATP-binding cassette (ABC) superfamily. We now show that the gene has two major transcripts of 4.5 and 1.3 kb. In breast cancer, normal breast, and testis, the MRP9 gene transcript is 4.5 kb in size and encodes a 100-kDa protein. The protein is predicted to have 8 instead of 12 membrane-spanning regions. When compared with closely related ABC family members, it lacks transmembrane domains 3, 4, 11, and 12 and the second nucleotide-binding domain. In other tissues including brain, skeletal muscle, and ovary, the transcript size is 1.3 kb. This smaller transcript encodes a nucleotide-binding protein of approximate to25 kDa in size. An in situ hybridization study shows that the 4.5-kb transcript is expressed in the epithelial cells of breast cancer. An antipeptide antibody designed to react with the amino terminus of the protein detects a 100-kDa protein in testis and the membrane fraction of a breast cancer cell line. Because the 4.5-kb RNA is highly expressed in breast cancer and not expressed at detectable levels in essential normal tissues, MRP9 could be a useful target for the immunotherapy of breast cancer. Because of the unusual topology of the two variants of MRP9, we speculate that they may have a different function from other family members. C1 NCI, Mol Biol Lab, Ctr Canc Res, Bethesda, MD 20892 USA. RP Pastan, I (reprint author), NCI, Mol Biol Lab, Ctr Canc Res, Bldg 37,Room 5106,37 Convent Dr,MSC 4264, Bethesda, MD 20892 USA. NR 23 TC 81 Z9 87 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 MAY 14 PY 2002 VL 99 IS 10 BP 6997 EP 7002 DI 10.1073/pnas.102187299 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300086 PM 12011458 ER PT J AU Valentin, A Rosati, M Patenaude, DJ Hatzakis, A Kostrikis, LG Lazanas, M Wyvill, KM Yarchoan, R Pavlakis, GN AF Valentin, A Rosati, M Patenaude, DJ Hatzakis, A Kostrikis, LG Lazanas, M Wyvill, KM Yarchoan, R Pavlakis, GN TI Persistent HIV-1 infection of natural killer cells in patients receiving highly active antiretroviral therapy SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; BLOOD MONONUCLEAR-CELLS; CD4(+) T-CELLS; MULTIDRUG TRANSPORTER; COMBINATION THERAPY; PROTEASE INHIBITORS; DISEASE PROGRESSION; HUMAN HERPESVIRUS-6; PLASMA VIREMIA; LYMPHOCYTES AB We have identified a subset of CD56(+)CD3(-) human natural killer (NK) cells that express CD4 and the HIV coreceptors CCR5 and CXCR4. These cells can be productively infected in vitro by both CCR5- and CXCR4-using molecular clones of HIV-1 in a CD4-dependent manner. Analysis of HIV-infected persons showed that viral DNA is present in purified NK cells, and virus could be rescued from these cells after in vitro cultivation. Longitudinal analysis of the HIV-1 DNA levels in NK cells from patients after 1-2 years of highly active antiretroviral therapy indicated that NK cells remain persistently infected and account for a substantial amount of the viral DNA in peripheral blood mononuclear cells. These results demonstrate that a subset of non-T cells with NK markers are persistently infected and suggest that HIV infection of NK cells is important for virus persistence. The properties of the virus reservoir in these cells should be considered in attempts to further optimize antiretroviral therapies. C1 NCI, Human Retrovirus Sect, Basic Res Lab, Frederick, MD 21702 USA. Univ Athens, Sch Med, GR-11527 Athens, Greece. Red Cross Gen Hosp, Athens 11527, Greece. NCI, HIV & AIDS Malignancy Branch, Bethesda, MD 20892 USA. RP Pavlakis, GN (reprint author), NCI, Human Retrovirus Sect, Basic Res Lab, Bldg 535,Room 210, Frederick, MD 21702 USA. RI Kostrikis, Leondios/A-5330-2016 OI Kostrikis, Leondios/0000-0002-5340-7109 NR 36 TC 103 Z9 107 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 MAY 14 PY 2002 VL 99 IS 10 BP 7015 EP 7020 DI 10.1073/pnas.102672999 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300089 PM 12011460 ER PT J AU Zouboulis, CC Seltmann, H Hiroi, N Chen, WC Young, M Oeff, M Scherbaum, WA Orfanos, CE McCann, SM Bornstein, SR AF Zouboulis, CC Seltmann, H Hiroi, N Chen, WC Young, M Oeff, M Scherbaum, WA Orfanos, CE McCann, SM Bornstein, SR TI Corticotropin-releasing hormone: An autocrine hormone that promotes lipogenesis in human sebocytes SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID BINDING PROTEIN; FACTOR CRF; CUTANEOUS EXPRESSION; STRESS-RESPONSE; HUMAN SKIN; RECEPTORS; CELLS; DIFFERENTIATION; HOMEOSTASIS; UROCORTIN AB Sebaceous glands may be involved in a pathway conceptually similar to that of the hypothalamic-pituitary-adrenal (HIPA) axis. Such a pathway has been described and may occur in human skin and lately in the sebaceous glands because they express neuropeptide receptors. Corticotropin-releasing hormone (CRH) is the most proximal element of the HPA axis, and it acts as central coordinator for neuroendocrine and behavioral responses to stress. To further examine the probability of an HPA equivalent pathway, we investigated the expression of CRH, CRH-binding protein (CRH-BP), and CRH receptors (CRH-R) in SZ95 sebocytes in vitro and their regulation by CRH and several other hormones. CRH, CRH-BP, CRH-R1, and CRH-R2 were detectable in SZ95 sebocytes at the mRNA and protein levels: CRH-R1 was the predominant type (CRH-R1/CRH-R2 = 2). CRH was biologically active on human sebocytes: it induced biphasic increase in synthesis of sebaceous lipids with a maximum stimulation at 10(-7) M and up-regulated mRNA levels of 3beta-hydroxysteroid dehydrogenase/Delta(5-4) isomerase, although it did not affect cell viability, cell proliferation, or IL-1beta-induced IL-8 release. CRH, dehydroepiandrosterone, and 17beta-estradiol did not modulate CRH-R expression, whereas testosterone at 10(-7) M down-regulated CRH-R1 and CRH-R2 mRNA expression at 6 to 24 h, and growth hormone (GH) switched CRH-R1 mRNA expression to CRH-R2 at 24 h. Based on these findings, CRH may be an autocrine hormone for human sebocytes that exerts homeostatic lipogenic activity, whereas testosterone and growth hormone induce CRH negative feedback. The findings implicate CRH in the clinical development of acne, seborrhea, androgenetic alopecia, skin aging, xerosis, and other skin disorders associated with alterations in lipid formation of sebaceous origin. C1 Free Univ Berlin, Univ Med Ctr Benjamin Franklin, Dept Dermatol, D-14195 Berlin, Germany. Univ Dusseldorf, Univ Med Ctr, Dept Endocrinol, D-40225 Dusseldorf, Germany. Natl Inst Child Hlth & Dev, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. Natl Cheng Kung Univ, Coll Med, Dept Dermatol, Tainan 704, Taiwan. Pennington Biomed Res Ctr, Baton Rouge, LA 70808 USA. RP Zouboulis, CC (reprint author), Free Univ Berlin, Univ Med Ctr Benjamin Franklin, Dept Dermatol, Fabeckstr 60-62, D-14195 Berlin, Germany. RI Dr. Zouboulis, Christos/I-4493-2013 OI Dr. Zouboulis, Christos/0000-0003-1646-2608 NR 36 TC 141 Z9 147 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 MAY 14 PY 2002 VL 99 IS 10 BP 7148 EP 7153 DI 10.1073/pnas.102180999 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 552TZ UT WOS:000175637300112 PM 12011471 ER PT J AU Walsh, CR Larson, MG Leip, ER Vasan, RS Levy, D AF Walsh, CR Larson, MG Leip, ER Vasan, RS Levy, D TI Serum potassium and risk of cardiovascular disease - The Framingham Heart Study SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID CREATININE ASSAYS; DIETARY POTASSIUM; STROKE MORTALITY; SODIUM; MAGNESIUM; MEN AB Background: Published studies of the association between serum potassium concentration and risk for cardiovascular disease in community-based populations have reported conflicting results. We sought to determine the association between serum potassium concentration and cardiovascular disease risk in the Framingham Heart Study. Methods: A total of 3151 participants (mean age, 43 years; 48% men) in the Framingham Heart Study who were free of cardiovascular disease and not taking medications affecting potassium homeostasis had serum potassium levels measured (1979-1983). Proportional hazards models were used to determine the association of serum potassium concentration at baseline with the incidence of cardiovascular disease at follow-up. Results: During mean follow-up of 16 years, 313 cardiovascular disease events occurred, including 46 cardiovascular disease-related deaths. After adjustment for age, serum potassium level was marginally associated with risk of cardiovascular disease (hazard ratio [HR] per I mg/dL increment, 1.03; 95% confidence interval [CI], 1.00-1.05; P=.02). However, after further adjustment for multiple confounders, serum potassium level was not significantly associated with cardiovascular disease risk (HR, 1.00; 95% Cl, 0.98-1.03). There were no significant associations between serum potassium level and cardiovascular disease-related death in either age- and sex-adjusted models (HR, 1.06; 95% Cl, 0.99-1.12) or multivariable-adjusted models (HR, 1.04; 95% CI, 0.97-1.11). Conclusion. In our community-based sample of individuals free of cardiovascular disease and not taking medications that affect potassium homeostasis, serum potassium level was not associated with risk of cardiovascular disease. C1 NHLBI, Framingham Heart Study, NIH, Framingham, MA 01702 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Med,Div Cardiol, Boston, MA 02115 USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med, Boston, MA 02115 USA. Boston Univ, Sch Med, Evans Dept Med, Sect Epidemiol & Prevent Med, Boston, MA 02118 USA. RP Levy, D (reprint author), NHLBI, Framingham Heart Study, NIH, 5 Thurber St, Framingham, MA 01702 USA. OI Ramachandran, Vasan/0000-0001-7357-5970 FU NHLBI NIH HHS [1K24 HL04334, N01-HC-38038] NR 21 TC 12 Z9 12 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD MAY 13 PY 2002 VL 162 IS 9 BP 1007 EP 1012 DI 10.1001/archinte.162.9.1007 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 550PW UT WOS:000175512300007 PM 11996610 ER PT J AU He, J Muntner, P Chen, J Roccella, EJ Streiffer, RH Whelton, PK AF He, J Muntner, P Chen, J Roccella, EJ Streiffer, RH Whelton, PK TI Factors associated with hypertension control in the general population of the United States SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID BLOOD-PRESSURE; INNER-CITY; HEALTH; CARE; PREVALENCE; MEN; AWARENESS; DISEASE; RISK AB Background: Uncontrolled hypertension is the most and important risk factor for cardiovascular and renal disease. We studied factors associated with hypertension control in the Third National Health and Nutrition Examination Survey. Methods: A total of 3077 non-Hispanic whites, 1742 non-Hispanic blacks, and 1067 Mexican Americans 18 years or older with hypertension were included in the current analysis. Blood pressure was measured by trained observers by means of a standard mercury sphygmomanometer, and controlled hypertension was defined as a mean systolic/diastolic blood pressure less than 140/90 mm Hg. Results: Percentages of persons with controlled hypertension differed significantly by ethnicity and sex: 1.9.2% and 28.7% for white men and women, 17.5% and 28.6% for black men and women, and 12.7% and 18.0% for Mexican American men and women, respectively. After adjustment for important covariables, percentages of persons with controlled hypertension were significantly higher among persons who were currently (odds ratio [OR] 2.39; 95% confidence interval 101, 1.52-3.74) or formerly (OR, 1.81; 95% CL 1.12-2.93) married, had private health insurance (OR, 1.59; 95% CI, 1.02-2.49), visited the same facility for their health care (OR, 2.77; 95% C1, 1.88-4.09) or saw the same provider for their health care (OR, 2.29; 95% C1, 1.74-3.02), had their blood pressure checked during the preceding 6 months (OR, 8.00; 95% C1, 3.75-17.1) or 6 to 11 months (OR, 5.31; 2.51-11.2), and reported using lifestyle modification to control their hypertension (OR, 6.02; 95% C1, 4.20-8.63). Conclusion: These data strongly suggest that access to a regular source of health care and modification of lifestyle are important factors in the control of hypertension in the community. C1 Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, New Orleans, LA 70112 USA. Tulane Univ, Sch Med, Dept Med, New Orleans, LA 70112 USA. Tulane Univ, Sch Med, Dept Family & Community Med, New Orleans, LA 70112 USA. NHLBI, Bethesda, MD 20892 USA. RP He, J (reprint author), Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, 1430 Tulane Ave SL18, New Orleans, LA 70112 USA. FU NHLBI NIH HHS [R01HL60300] NR 23 TC 146 Z9 156 U1 0 U2 3 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 MAY 13 PY 2002 VL 162 IS 9 BP 1051 EP 1058 DI 10.1001/archinte.162.9.1051 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 550PW UT WOS:000175512300014 PM 11996617 ER PT J AU Shi, PA Hematti, P von Kalle, C Dunbar, CE AF Shi, PA Hematti, P von Kalle, C Dunbar, CE TI Genetic marking as an approach to studying in vivo hematopoiesis: progress in the non-human primate model SO ONCOGENE LA English DT Article DE hematopoietic stem cells; retroviral vectors; in vivo gene tracking; clonality ID STEM-CELL FACTOR; COLONY-STIMULATING FACTOR; MARROW REPOPULATING CELLS; X-CHROMOSOME INACTIVATION; RHESUS PERIPHERAL-BLOOD; LONG-TERM ENGRAFTMENT; IMMUNE-DEFICIENT MICE; BONE-MARROW; NONHUMAN-PRIMATES; IN-VIVO AB Retroviral insertion site analysis following transplantation of marked hematopoietic stem cells (HSCs) is a powerful method for studying hematopoiesis in vivo. High-level gene transfer efficiency was achieved in murine models in the late 1980s, but early human gene transfer protocols into hematopoietic stem and progenitor cells using the murine methodology showed consistently poor results. The utility of non-human primates as pre-clinical models has since become apparent. Modifications in retroviral transduction conditions have resulted in stable long-term gene transfer efficiency as high as 15-20% to primitive repopulating cells in non-human primate models. This has permitted, for the first time in a large animal model, tracking of individual stem and progenitor cell clones via insertion site analysis, an advantage over competitive transplantation studies, which cannot firmly evaluate the number or life span of individual clones contributing to hematopoiesis. Retroviral tracking studies in mice suggest that stable hematopoiesis may be dominated by a small number of clones, but these studies have been limited by insensitive detection methods, low numbers of transplanted stem cells, and limited life span of immunodeficient mice. Autologous transplantation studies in non-human primates have just begun and have the potential to shed light on controversial issues such as the number of clones contributing to stable hematopoiesis, clonal succession, and lineage commitment, as well as the effect of clinically relevant manipulations such as cytokines, chemotherapy, and radiation on hematopoiesis. These approaches will have significant impact in studying various aspects of stem cell biology including the phenomenon of stem cell plasticity. C1 NHLBI, Mol Hematopoiesis Sect, Hematol Branch, NIH, Bethesda, MD 20892 USA. Univ Freiburg, Dept Internal Med, D-7800 Freiburg, Germany. RP Dunbar, CE (reprint author), NHLBI, Mol Hematopoiesis Sect, Hematol Branch, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 86 TC 20 Z9 20 U1 1 U2 2 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 13 PY 2002 VL 21 IS 21 BP 3274 EP 3283 DI 10.1038/sj/onc/1205320 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 552RL UT WOS:000175633300004 PM 12032769 ER PT J AU Rane, SG Reddy, EP AF Rane, SG Reddy, EP TI JAKs, STATs and Src kinases in hematopoiesis SO ONCOGENE LA English DT Review DE hematopoiesis; Src kinases; JAK kinases; STATs; ras; PI3K ID COLONY-STIMULATING FACTOR; PROTEIN-TYROSINE KINASE; GROWTH-HORMONE RECEPTOR; DNA-BINDING ACTIVITY; COMMON BETA-SUBUNIT; SEVERE COMBINED IMMUNODEFICIENCY; SIGNAL-TRANSDUCTION PATHWAYS; LEUKEMIA INHIBITORY FACTOR; PROGENITOR-CELL LINE; MICE LACKING JAK3 AB Hematopoiesis is the cumulative result of intricately regulated signal transduction cascades that are mediated by cytokines and their cognate receptors. Proper culmination of these diverse signaling pathways forms the basis for an orderly generation of different cell types and aberrations in these pathways is an underlying cause for diseases such as leukemias and other myeloproliferative and lympho proliferative disorders. Over the past decade, downstream signal transduction events initiated upon cytokine/growth factor stimulation have been a major focus of basic and applied biomedical research. As a result, several key concepts have emerged allowing a better understanding of the complex signaling processes. A group of transcription factors, termed signal transducers and activators of transcription (STATs) appear to orchestrate the downstream events propagated by cytokine/growth factor interactions with their cognate receptors. Similarly, cytoplasmic Janus protein tyrosine kinases (JAKs) and Src family of kinases seem to play a critical role in diverse signal transduction pathways that govern cellular survival, proliferation, differentiation and apoptosis. Accumulating evidence suggests that STAT protein activation may be mediated by members of both JAK and Src family members following cytokine/growth factor stimulation. In addition, JAK kinases appear to be essential for the phosphorylation of the cytokine receptors which results in the creation of docking sites on the receptors for binding of SH2-containing proteins such as STATs, Src-kinases and other signaling intermediates. Cell and tissue-speciticity of cytokine action appears to be determined by the nature of signal transduction pathways activated by cytokine/receptor interactions. The integration of these diverse signaling cues from active JAK kinases, members of the Src-family kinases and STAT proteins, leads to cell proliferation, cell survival and differentiation, the end-point of the cytokine/growth factor stimulus. C1 Temple Univ, Sch Med, Fels Inst Canc Res & Mol Biol, Philadelphia, PA 19140 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Reddy, EP (reprint author), Temple Univ, Sch Med, Fels Inst Canc Res & Mol Biol, 3307 N Broad St, Philadelphia, PA 19140 USA. EM reddy@unix.temple.edu RI Reddy, E. Premkumar/F-6233-2011 FU NIEHS NIH HHS [ES09225] NR 253 TC 155 Z9 168 U1 5 U2 13 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 13 PY 2002 VL 21 IS 21 BP 3334 EP 3358 DI 10.1038/sj/onc/1205398 PG 25 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 552RL UT WOS:000175633300008 PM 12032773 ER PT J AU Polis, MA Sidorov, IA Dimitrov, DS AF Polis, MA Sidorov, IA Dimitrov, DS TI Early assessment of anti-HIV drug efficacy - Reply SO LANCET LA English DT Letter C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NCI, Lab Expt & Computat Biol, NIH, Frederick, MD 21701 USA. RP Polis, MA (reprint author), NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NR 2 TC 0 Z9 0 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 MAY 11 PY 2002 VL 359 IS 9318 BP 1697 EP 1697 DI 10.1016/S0140-6736(02)08572-0 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 551ZF UT WOS:000175593300041 ER PT J AU Flugy, AM D'Amato, M Russo, D Di Bella, MA Alaimo, G Kohn, EC De Leo, G Alessandro, R AF Flugy, AM D'Amato, M Russo, D Di Bella, MA Alaimo, G Kohn, EC De Leo, G Alessandro, R TI E-selectin modulates the malignant properties of T84 colon carcinoma cells SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE E-selectin signaling; metastasis; colon carcinoma; tumor cells; endothelial cells ID TUMOR-CELLS; ENDOTHELIAL-CELLS; MATRIX METALLOPROTEINASES; TYROSINE KINASES; GROWTH-FACTOR; SIALYL-LEWIS; CANCER-CELLS; METASTASIS; EXPRESSION; ADHESION AB The extravasation of metastatic cells is regulated by molecular events involving the initial adhesion of tumor cells to the endothelium and subsequently the migration of cells in the host connective tissue. E-selectin on endothelial cells and sialyl Lewis X carbohydrate component on tumor cells are mainly involved in the adhesion of colon carcinoma cells to the endothelium of target organ. Interaction of T84 colon cancer cells to purified E-selectin in vitro caused an increase in the tyrosine phosphorylation of a number of proteins as well as the modulation of cellular properties correlated to the metastatic phenotype. Specifically, E-selectin-stimulated actin reorganization, increased collagenase secretion, and induced cell migration. Treatment of T84 cells with herbimycin A inhibited cell adhesion as well as selectin-induced increase of cell migration, and cytoskeleton assembly. Our data demonstrate that binding of cancer cells to E-selectin starts signal transduction pathways which may affect the tumor metastatic abilities. (C) 2002 Elsevier Science (USA). All rights reserved. C1 Univ Palermo, Dept Biopathol & Biomed Methodol, Sect Biol & Genet, Palermo, Italy. NCI, Mol Signalling Sect, Pathol Lab, Bethesda, MD 20892 USA. RP Alessandro, R (reprint author), Univ Palermo, Dept Biopathol & Biomed Methodol, Sect Biol & Genet, Palermo, Italy. NR 30 TC 18 Z9 19 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 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 10 PY 2002 VL 293 IS 3 BP 1099 EP 1106 AR PII S0006-291X(02)00337-6 DI 10.1016/S0006-291X(02)00337-6 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 552VM UT WOS:000175640800033 PM 12051773 ER PT J AU Leyton, J Garcia-Marin, L Jensen, RT Moody, TW AF Leyton, J Garcia-Marin, L Jensen, RT Moody, TW TI Neurotensin causes tyrosine phosphorylation of focal adhesion kinase in lung cancer cells SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE lung cancer; focal adhesion kinase; neurotensin; SR48692; proliferation ID SWISS 3T3 CELLS; RECEPTOR ANTAGONIST; PROSTATE-CANCER; BETA-ENDORPHIN; GROWTH; PAXILLIN; BOMBESIN; PROTEIN; EXPRESSION; PEPTIDES AB The effects of neurotensin on focal adhesion kinase were investigated using lung cancer cells. Neurotensin bound with high affinity to large cell carcinoma cell line NCI-H1299 as did neurotensin-(8-13), but not neurotensin-(1-7) or levocabastine. Addition of 100 nM neurotensin to NCI-H1299 cells caused transient tyrosine phosphorylation of focal adhesion kinase which was maximal after 1-2.5 min. Also, neurotensin-(8-13), but not neurotensin-(1-8) or levocabastine, caused tyrosine phosphorylation of focal adhesion kinase after addition to NCI-H1299 cells. Focal adhesion kinase tyrosine phosphorylation caused by neurotensin was inhibited by the nonpeptide neurotensin receptor antagonist (2-(1-(7-chloroquinolin-4-yl)-5-(2,6-dimethoxyphenyl)-1H-pyrazole-3-carbonyl)amino)-adamantane-2-carboxylic acid) (SR48692). SR48692 inhibited the clonal growth of NCI-H1299 cells, whereas neurotensin-stimulated proliferation and levocabastine had no effect. These results indicate that lung cancer cells have functional neurotensin receptors which regulate focal adhesion kinase tyrosine phosphorylation, It remains to be determined if neurotensin receptors and focal adhesion kinase plays a role in lung cancer cellular adhesion and migration. (C) 2002 Published by Elsevier Science B.V. C1 Cell Canc Biol Dept, Med Branch, Rockville, MD 20850 USA. NIDDKD, Digest Dis Branch, Bethesda, MD 20892 USA. RP Moody, TW (reprint author), Cell Canc Biol Dept, Med Branch, NCI Bldg,KWC,Rm 300,9610 Med Ctr Dr, Rockville, MD 20850 USA. RI Garcia-Marin, Luis /L-4680-2014 OI Garcia-Marin, Luis /0000-0002-1795-7381 NR 52 TC 26 Z9 28 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD MAY 10 PY 2002 VL 442 IS 3 BP 179 EP 186 AR PII S0014-2999(02)01539-X DI 10.1016/S0014-2999(02)01539-X PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 567KV UT WOS:000176484700002 PM 12065070 ER PT J AU Schindler, CW Bross, JG Thorndike, EB AF Schindler, CW Bross, JG Thorndike, EB TI Gender differences in the behavioral effects of methamphetamine SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE methamphetamine; locomotor activity; place preference; sex; (rat) ID SEX-DIFFERENCES; GONADAL-HORMONES; RATS; COCAINE; DOPAMINE; FEMALE; SENSITIZATION; AMPHETAMINE; SENSITIVITY; RECEPTOR AB The effects of methamphetamine were tested in male and female rats on two different behavioral tasks. Following habituation to a locomotor activity chamber, female rats were more sensitive to the locomotor activating effect of i.p. methamphetamine (0.1-3.0 mg/kg) than were male rats. A similar effect has been observed for other psychomotor stimulants, including cocaine and amphetamine. However, males and females did not differ on methamphetamine-induced place preference following eight conditioning trials with a wide range of doses (0.1-5.6 mg/kg). These results suggest that males and females differ in their response to methamphetamine for only some behavioral tasks. Published by Elsevier Science B.V. C1 NIDA, Preclin Pharmacol Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. RP Schindler, CW (reprint author), NIDA, Preclin Pharmacol Sect, Intramural Res Program, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 25 TC 39 Z9 40 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD MAY 10 PY 2002 VL 442 IS 3 BP 231 EP 235 AR PII S0014-2999(02)01550-9 DI 10.1016/S0014-2999(02)01550-9 PG 5 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 567KV UT WOS:000176484700008 PM 12065076 ER PT J AU Brenner, AV Linet, MS Fine, HA Shapiro, WR Selker, RG Black, PM Inskip, PD AF Brenner, AV Linet, MS Fine, HA Shapiro, WR Selker, RG Black, PM Inskip, PD TI History of allergies and autoimmune diseases and risk of brain tumors in adults SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE case-control studies; brain neoplasms; glioma; meningioma; neuroma, acoustic; adult; risk factors; hypersensitivity; autoimmune diseases ID CENTRAL-NERVOUS-SYSTEM; RHEUMATOID-ARTHRITIS; CANCER INCIDENCE; UNITED-STATES; MULTIPLE-SCLEROSIS; DIABETES-MELLITUS; B-CELLS; T-CELLS; 2 PARTS; POPULATION AB To explore a possible influence of the immune system in the development of brain tumors, we evaluated the relationship between history of allergies and autoimmune diseases and risk of brain tumors within a large, hospital-based case-control study. Cases (n 782) were patients recently diagnosed with glioma (n 489), meningioma (n = 197) or acoustic neuroma (n = 96) at hospitals in Boston, Phoenix and Pittsburgh (USA). Controls (n = 799) were patients hospitalized for a variety of nonmalignant conditions and frequency-matched to cases by hospital, age, sex, race/ethnicity and distance of residence from hospital. Research nurses collected data by personal interview of patients. Odds ratios (OR) and 95% confidence intervals (Cl) were calculated using unconditional logistic regression. There was a significant inverse association between glioma and history of any allergies (OR = 0.67, 95% Cl = 0.52-0.86) or autoimmune diseases (OR = 0.49, 95% Cl = 0.35-0.69). No significant associations were evident for meningioma or acoustic neuroma with history of any allergies. An inverse association was observed between meningioma and history of autoimmune diseases (OR = 0.59, 95% CI = 0.38-0.92). There was a suggestion of interaction between allergies and autoimmune diseases on risk of glioma (p = 0.06), with subjects having both conditions being at lowest risk (OR = 0.24, 95% Cl = 0.14-0.42). Among the specific conditions, asthma and diabetes showed the most consistent associations (OR = 0.63, 95% Cl = 0.43-0.92 and OR = 0.44, 95% Cl = 0.27-0.70, respectively). Our results add to evidence that persons with allergies or autoimmune diseases are at reduced risk of glioma. The basis of the associations is not clear, but they might imply a role of immunologic factors in the development of brain tumors. Published 2002 Wiley-Liss, Inc.(dagger) C1 NCI, Div Canc Epidemiol & Genet, Radiat Epidemiol Branch, Bethesda, MD 20892 USA. NCI, Neurooncol Branch, Bethesda, MD 20892 USA. St Josephs Hosp, Barrow Neurol Inst, Phoenix, AZ 85013 USA. Med Ctr, Phoenix, AZ USA. Western Penn Hosp, Div Neurosurg, Pittsburgh, PA 15224 USA. Brigham & Womens Hosp, Boston, MA 02115 USA. RP Brenner, AV (reprint author), NCI, Div Canc Epidemiol & Genet, Radiat Epidemiol Branch, 6120 Execut Blvd,MSC 7362, Bethesda, MD 20892 USA. NR 53 TC 128 Z9 134 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 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD MAY 10 PY 2002 VL 99 IS 2 BP 252 EP 259 DI 10.1002/ijc.10320 PG 8 WC Oncology SC Oncology GA 546XE UT WOS:000175301000014 PM 11979441 ER PT J AU Kostic, TS Tomic, M Andric, SA Stojilkovic, SS AF Kostic, TS Tomic, M Andric, SA Stojilkovic, SS TI Calcium-independent and cAMP-dependent modulation of soluble guanylyl cyclase activity by G protein-coupled receptors in pituitary cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NITRIC-OXIDE SYNTHASE; INTRACELLULAR CA2+ CONCENTRATION; CORTICOTROPIN-RELEASING FACTOR; PURIFIED RAT SOMATOTROPHS; GROWTH-HORMONE SECRETION; KINASE-C; PHOSPHORYLATION; EXPRESSION; GLAND; GONADOTROPHS AB It is well established that G protein-coupled receptors stimulate nitric oxide-sensitive soluble guanylyl cyclase by increasing intracellular Ca2+ and activating Ca2+-dependent nitric-oxide synthases. In pituitary cells receptors that stimulated adenylyl cyclase, growth hormone-releasing hormone, corticotropin-releasing factor, and thyrotropin-releasing hormone also stimulated calcium signaling and increased cGMP levels, whereas receptors that inhibited adenylyl cyclase, endothelin-A, and dopamine-2 also inhibited spontaneous calcium transients and decreased cGMP levels. However, receptor-controlled up- and down-regulation of cyclic nucleotide accumulation was not blocked by abolition of Ca2+ signaling, suggesting that cAMP production affects cGMP accumulation. Agonist-induced cGMP accumulation was observed in cells incubated in the presence of various phosphodiesterase and soluble guanylyl cyclase inhibitors, confirming that G(s)-coupled receptors stimulated de novo cGMP production. Furthermore, cholera toxin (an activator of G(s)), forskolin (an activator of adenylyl cyclase), and 8-Br-cAMP (a permeable cAMP analog) mimicked the stimulatory action of G(s)-coupled receptors on cGMP production. Basal, agonist-, cholera toxin-, and forskolin-stimulated cGMP production, but not cAMP production, was significantly reduced in cells treated with H89, a protein kinase A inhibitor. These results indicate that coupling seven plasma membrane-domain receptors to an adenylyl cyclase signaling pathway provides an additional calcium-independent and cAMP-dependent mechanism for modulating soluble guanylyl cyclase activity in pituitary cells. C1 NICHHD, Sect Cellular Signaling, ERRB, NIH, Bethesda, MD 20892 USA. RP Stojilkovic, SS (reprint author), NICHHD, Sect Cellular Signaling, ERRB, NIH, Bldg 49,Room 6A-36,49 convent Dr, Bethesda, MD 20892 USA. RI Tomic, Melanija/C-3371-2016 NR 54 TC 18 Z9 19 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 MAY 10 PY 2002 VL 277 IS 19 BP 16412 EP 16418 DI 10.1074/jbc.M112439200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551MB UT WOS:000175564500011 PM 11867632 ER PT J AU Wick, MJ Wick, KR Chen, H He, HL Dong, LQ Quon, MJ Liu, F AF Wick, MJ Wick, KR Chen, H He, HL Dong, LQ Quon, MJ Liu, F TI Substitution of the autophosphorylation site Thr(516) with a negatively charged residue confers constitutive activity to mouse 3-phosphoinositide-dependent protein kinase-1 in cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PHOSPHOINOSITIDE-DEPENDENT KINASE-1; C-ZETA; ACTIVATION; PHOSPHORYLATION; INSULIN; MECHANISM; PDK-1; TRANSLOCATION; SERUM; LOCALIZATION AB 3-Phosphoinositide-dependent protein kinase-1 (PDK-1) is a serine/threonine kinase that has been found to phosphorylate and activate several members of the AGC protein kinase family including protein kinase B (Akt), p70 S6 kinase, and protein kinase Czeta. However, the mechanism(s) by which PDK-1 is regulated remains unclear. Here we show that mouse PDK-1 (mPDK-1) undergoes autophosphorylation in vitro on both serine and threonine residues. In addition, we have identified Ser(399) and Thr(516) as the major mPDK-1 autophosphorylation sites in vitro. Furthermore, we have found that these two residues, as well as Ser(244) in the activation loop, are phosphorylated in cells and demonstrated that Ser(244) is a major in vivo phosphorylation site. Abolishment of phosphorylation at Ser(244), but not at Ser(399) or Thr(516), led to a significant decrease of mPDK-1 autophosphorylation and kinase activity in vitro, indicating that autophosphorylation at Ser(399) or Thr(516) is not essential for mPDK-1 autokinase activity. However, overexpression of mPDK-1(T516E), but not of mPDK-1(S244E) or mPDK-1(S399D), in Chinese hamster ovary and HEK293 cells was sufficient to induce Akt phosphorylation at Thr(308) to a level similar to that of insulin stimulation. Furthermore, this increase in phosphorylation was independent of the Pleckstrin homology domain of Akt. Taken together, our results suggest that mPDK-1 undergoes autophosphorylation at multiple sites and that this phosphorylation may be essential for PDK-1 to interact with and phosphorylate its downstream substrates in vivo. C1 Univ Texas, Hlth Sci Ctr, Dept Pharmacol, San Antonio, TX 78229 USA. Univ Texas, Hlth Sci Ctr, Dept Biochem, San Antonio, TX 78229 USA. NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Liu, F (reprint author), Univ Texas, Hlth Sci Ctr, Dept Pharmacol, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA. RI Quon, Michael/B-1970-2008; OI Quon, Michael/0000-0002-9601-9915; Quon , Michael /0000-0002-5289-3707 FU NIA NIH HHS [2T32AG00205-11]; NIDDK NIH HHS [DK56166] NR 31 TC 33 Z9 34 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 10 PY 2002 VL 277 IS 19 BP 16632 EP 16638 DI 10.1074/jbc.M112402200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551MB UT WOS:000175564500040 PM 11877406 ER PT J AU Kvaratskhelia, M Budihas, SR Le Grice, SFJ AF Kvaratskhelia, M Budihas, SR Le Grice, SFJ TI Pre-existing distortions in nucleic acid structure aid polypurine tract selection by HIV-1 reverse transcriptase SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; SEQUENCE FEATURES IMPORTANT; MURINE LEUKEMIA-VIRUS; STRAND DNA-SYNTHESIS; RNA-POLYMERASE; CRYSTAL-STRUCTURE; RESOLVING ENZYME; H DOMAIN; COMPLEX; INITIATION AB Precise cleavage at the polypurine tract (PPT)/U3 junction by human immunodeficiency virus type 1 (HIV-1) reverse transcriptase RNase H is critical for generating a correct viral DNA end for subsequent integration. Using potassium permanganate (KMnO4) modification, we have identified a significant distortion in the nucleic acid structure at the HIV-1 PPT/U3 junction in the absence of trans-acting factors. Unusually high reactivity of template thymine +1 is detected when the PPT primer is extended by DNA or RNA at its 3' terminus. Chemical footprinting suggests that the extent of base unstacking in the wild-type species is comparable when the +1 A:T base pair is replaced by a C:T mismatch. However, reactivity of this template base is diminished after alterations to upstream (rA)(4):(dT)(4) or (rG)(6):(dC)(6) tracts. Importantly, there is a correlation between the structural deformation at base pair +1 and precise cleavage at the PPT/U3 junction by HIV-1 reverse transcriptase/RNase H. KMnO4 modification also revealed unusually high reactivity for one of two (dT)(4): (rA)(4) duplexes upstream of the PPT/U3 junction, suggesting a significant structural distortion within the PPT itself in the absence of the retroviral polymerase. Structural abnormalities in this region are not only essential for resistance of the PPT to hydrolysis but also significantly impact the conformation of the PPT/U3 junction. Our data collectively suggest that the entire PPT sequence contributes to the structural distortion at the PPT/U3 junction, potentially providing a mechanism for its selective processing. C1 NCI Frederick, Reverse Transcriptase Biochem Sect, Resistance Mech Lab, HIV Drug Resistance Program,NIH, Frederick, MD 21702 USA. RP Le Grice, SFJ (reprint author), NCI Frederick, Reverse Transcriptase Biochem Sect, Resistance Mech Lab, HIV Drug Resistance Program,NIH, 535 Sultan St, Frederick, MD 21702 USA. NR 37 TC 45 Z9 46 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 MAY 10 PY 2002 VL 277 IS 19 BP 16689 EP 16696 DI 10.1074/jbc.M109914200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551MB UT WOS:000175564500047 PM 11875059 ER PT J AU Johnson, EN Druey, KM AF Johnson, EN Druey, KM TI Functional characterization of the G protein regulator RGS13 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GTPASE-ACTIVATING PROTEINS; SIGNALING RGS; PHOSPHOLIPASE-C; ALPHA-SUBUNITS; HUMAN GENE; RAT-BRAIN; EXPRESSION; GAIP; LOCALIZATION; RECEPTOR AB The signaling cascades evoked by G protein-coupled receptors are a predominant mechanism of cellular communication. The regulators of G protein signaling (RGS) comprise a family of proteins that attenuate G protein-mediated signal transduction. Here we report the characterization of RGS13, the smallest member of the RGS family, which has been cloned from human lung. RGS13 has been found most abundantly in human tonsil, followed by thymus, lung, lymph node, and spleen. RGS13 is a GTPase-activating protein for Galpha(i) and Galpha(o) but not Galpha(s). RGS13 binds Galpha(q) in the presence of aluminum magnesium fluoride, suggesting that it bears GTPase-activating protein activity toward Galpha(q). RGS13 blocks MAPK activity induced by Ga-i- or Galpha(q)-coupled receptors. RGS13 also attenuates GTPase-deficient Galpha(q) (Galpha(q)QL) mediated cAMP response element activation but not transcription evoked by constitutively active Galpha(12) or Galpha(13). Surprisingly, RGS13 inhibits cAMP generation elicited by stimulation of the beta(2)-adrenergic receptor. These data suggest that RGS13 may regulate Galpha(i)-, Galpha(q)-, and Galpha(s)-coupled signaling cascades. C1 NIAID, Mol Signal Transduct Sect, Lab Allerg Dis, NIH, Rockville, MD 20852 USA. RP Druey, KM (reprint author), 12441 Parklawn Dr,Rm 200E, Rockville, MD 20852 USA. NR 37 TC 34 Z9 34 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 MAY 10 PY 2002 VL 277 IS 19 BP 16768 EP 16774 DI 10.1074/jbc.M200751200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551MB UT WOS:000175564500057 PM 11875076 ER PT J AU Taniura, S Kamitani, H Watanabe, T Eling, TE AF Taniura, S Kamitani, H Watanabe, T Eling, TE TI Deacetylase inhibitors in normal human astrocyte cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROSTAGLANDIN ENDOPEROXIDE SYNTHASE; SIGNAL-TRANSDUCTION PATHWAY; HUMAN COLORECTAL-CARCINOMA; HISTONE DEACETYLASE; GROWTH-FACTOR; SP1 SITES; TRANSCRIPTIONAL REPRESSION; SODIUM-BUTYRATE; G/H SYNTHASE-2; GENE PROMOTER AB While cyclooxygenase (COX)-2 is a highly inducible gene, COX-1 is widely known as a noninducible gene and is constitutively expressed in a variety of cell lines and human tissues. Recently, several reports have indicated that COX-1 is also regulated at the transcriptional level by various stimuli. We present evidence that histone deacetylase (HDAC) inhibitors induce COX-1 transcription and translation in normal human astrocyte (NHA) cells and glioma cell lines. HDAC inhibitors increased acetylated histone H4 protein expression in NHA cells. The levels of COX-1 mRNA and protein were maximal at 24 and 48 h, respectively, after treatment with the specific HDAC inhibitor, trichostatin A (TSA). In addition, TSA-treated NHA cells produced prostaglandin E-2 as determined by enzyme-linked immunosorbent assay after incubation with 10 muM exogenous arachidonic acid, indicating that the induced COX-1 is functionally active. In addition to NHA cells, this up-regulation of COX-1 after treatment with HDAC inhibitors was observed in 5 different glioma cell lines. The nucleotide sequence of the inducible COX-1 cDNA was confirmed identical to human COX-1 that was previously reported. HDAC inhibitors stimulated COX-1 promoter activity as measured by luciferase reporter assays, suggesting that the induction of COX-1 is regulated at the transcriptional level. Furthermore, mutation analysis of the COX-1 promoter suggests that TSA-responsive element exists in the proximal Sp1-binding site at +25 to +31. In conclusion, COX-1 is an inducible gene in glial-derived cells including immortalized cells, and appears to be transcriptionally regulated by a unique mechanism associated with histone acetylation. C1 NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. Tottori Univ, Fac Med, Inst Neurol Sci, Div Neurosurg, Yonago, Tottori 6838504, Japan. RP Eling, TE (reprint author), NIEHS, Mol Carcinogenesis Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 47 TC 57 Z9 58 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 MAY 10 PY 2002 VL 277 IS 19 BP 16823 EP 16830 DI 10.1074/jbc.M200527200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 551MB UT WOS:000175564500064 PM 11877441 ER PT J AU Collins, PL Murphy, BR AF Collins, PL Murphy, BR TI Respiratory syncytial virus: Reverse genetics and vaccine strategies SO VIROLOGY LA English DT Review ID FORMALIN-INACTIVATED RSV; ENHANCED PULMONARY HISTOPATHOLOGY; CELL-MEDIATED-IMMUNITY; CD8(+) T-LYMPHOCYTES; REPLICATION IN-VITRO; BALB/C MICE; VIRAL REPLICATION; SUBUNIT VACCINE; G GLYCOPROTEIN; MESSENGER-RNA C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Collins, PL (reprint author), NIAID, Infect Dis Lab, NIH, Bldg 50,Room 6503,50 South Dr,MSC 8007, Bethesda, MD 20892 USA. NR 58 TC 51 Z9 52 U1 0 U2 1 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD MAY 10 PY 2002 VL 296 IS 2 BP 204 EP 211 DI 10.1006/viro.2002.1437 PG 8 WC Virology SC Virology GA 568HY UT WOS:000176537400002 PM 12069519 ER PT J AU Mohan, KVK Ghebrehiwet, B Atreya, CD AF Mohan, KVK Ghebrehiwet, B Atreya, CD TI The N-terminal conserved domain of rubella virus capsid interacts with the C-terminal region of cellular p32 and overexpression of p32 enhances the viral infectivity SO VIRUS RESEARCH LA English DT Article DE immunofluorescence; infectivity; overexpression; p32; rubella virus capsid; yeast two-hybrid ID GLOBULAR HEADS; BINDING-PROTEIN; C1Q; RECEPTOR; GC1Q-R; P-32; IDENTIFICATION; MITOCHONDRIA; LOCALIZATION; EXPRESSION AB Cellular 'defense collagens' are produced to launch virus-specific responses to clear the invading viruses, Cellular p32. the C1q binding protein is one such protein. In this report. we identified the interaction of p32 derived from a human lung diploid cell line (WI-38) with rubella virus capsid (RVCP from Therien strain) N-terminal 28-amino acid domain, which is conserved among several RV strains including the vaccine strains. We further identified that the C-terminal 69 aa of the mature p32 is sufficient to interact with the CP. In addition, we observed that in three independent Vero 76-derived cell lines constitutively overexpressing p32. the RV infectivity was enhanced. Our results suggest that RV has evolved a strategy whereby one of its proteins is recruited to interact with, and exploit the cellular defense machinery to its advantage. (C) 2002 Elsevier Science B.V. All rights reserved. C1 US FDA, Ctr Biol Evaluat & Res, Lab Pediat & Resp Viral Dis, Div Viral Prod,Sect viral Pathogenesis & Adverse, Bethesda, MD 20892 USA. SUNY Stony Brook, Dept Med, Stony Brook, NY 11794 USA. SUNY Stony Brook, Dept Pathol, Stony Brook, NY 11794 USA. RP Atreya, CD (reprint author), US FDA, Ctr Biol Evaluat & Res, Lab Pediat & Resp Viral Dis, Div Viral Prod,Sect viral Pathogenesis & Adverse, Bldg 29A,Room 2C-11,HFM-460,NIH Campus,8800 Rockv, Bethesda, MD 20892 USA. NR 33 TC 30 Z9 31 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0168-1702 J9 VIRUS RES JI Virus Res. PD MAY 10 PY 2002 VL 85 IS 2 BP 151 EP 161 AR PII S0168-1702(02)00030-8 DI 10.1016/S0168-1702(02)00030-8 PG 11 WC Virology SC Virology GA 570FV UT WOS:000176649000004 PM 12034482 ER PT J AU Ravi, RG Kim, HS Servos, J Zimmermann, H Lee, K Maddileti, S Boyer, JL Harden, TK Jacobson, KA AF Ravi, RG Kim, HS Servos, J Zimmermann, H Lee, K Maddileti, S Boyer, JL Harden, TK Jacobson, KA TI Adenine nucleotide analogues locked in a northern methanocarba conformation: Enhanced stability and potency as P2Y(1) receptor agonists SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID ECTO-ATP DIPHOSPHOHYDROLASE; PROTEIN-COUPLED RECEPTOR; GATED ION-CHANNEL; PURINERGIC RECEPTOR; BIOLOGICAL-ACTIVITY; P-2Y-PURINOCEPTOR AGONISTS; HETEROLOGOUS EXPRESSION; PLATELET-AGGREGATION; ADENOSINE RECEPTOR; PHOSPHOLIPASE-C AB Preference for the Northern (N) ring conformation of the ribose moiety of nucleotide 5'-triphosphate agonists at P2Y(1), P2Y(2), P2Y(4), and P2Y(11) receptors, but not P2Y(6) receptors, was established using a ring-constrained methanocarba (a 3.1.0-bicyclohexane) ring as a ribose substitute (Kim et al. J. Med. Chem. 2002, 45, 208-218.). We have now combined the ring-constrained (N)-methanocarba modification of adenine nucleotides with other functionalities known to enhance potency at P2 receptors. The potency of the newly synthesized analogues was determined in the stimulation of phospholipase C through activation of turkey erythrocyte P2Y, or human P2Y, and P2Y2 receptors stably expressed in astrocytoma cells. An (N)methanocarba-2-methylthio-ADP analogue displayed an EC50 at the hP2Y(1) receptor of 0.40 nM and was 55-fold more potent than the corresponding triphosphate and 16-fold more potent than the riboside 5'-diphosphate. 2-Cl-(N)-methanocarba-ATP and its N-6-Me analogue were also highly selective, full agonists at P2Y, receptors. The (N)-methanocarba-2-methylthio and 2-chloromonophosphate analogues were full agonists exhibiting micromolar potency at P2Y(1) receptors, while the corresponding ribosides were inactive. Although beta,y-methylene-ATP was inactive at P2Y receptors beta,y-methylene-(N)-methanocarba-ATP was a potent hP2Y1 receptor agonist with an EC50 of 160 nM and was selective versus hP2Y(2) and hP2Y(4) receptors. The rates of hydrolysis of Northern (N) and Southern (S) methanocarba analogues of AMP by rat 5'-ectonucleotidase were negligible. The rates of hydrolysis of the corresponding triphosphates by recombinant rat NTPDase 1 and 2 were studied. Both isomers were hydrolyzed by NTPDase 1 at about half the rate of ATP hydrolysis. The (N) isomer was hardly hydrolyzed by NTPDase 2, while the (S) isomer was hydrolyzed at one-third of the rate of ATP hydrolysis. This suggests that new, more stable and selective nucleotide agonists may be designed on the basis of the (N)-conformation, which greatly enhanced potency at P2Y, receptors. C1 NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Univ N Carolina, Sch Med, Dept Pharmacol, Chapel Hill, NC 27599 USA. Univ Frankfurt, Biozentrum, AK Neurochem, D-60439 Frankfurt, Germany. RP Jacobson, KA (reprint author), NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bldg 8A,Room B1A-17, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 FU Intramural NIH HHS [ZIA DK031116-22]; NHLBI NIH HHS [HL 54889, R01 HL054889, R29 HL054889]; NIGMS NIH HHS [GM 38213, R01 GM038213] NR 53 TC 45 Z9 45 U1 0 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD MAY 9 PY 2002 VL 45 IS 10 BP 2090 EP 2100 DI 10.1021/jm010538v PG 11 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 550DT UT WOS:000175488900015 PM 11985476 ER PT J AU Presley, JF Ward, TH Pfeifer, AC Siggia, ED Phair, RD Lippincott-Schwartz, J AF Presley, JF Ward, TH Pfeifer, AC Siggia, ED Phair, RD Lippincott-Schwartz, J TI Dissection of COPI and Arf1 dynamics in vivo and role in Golgi membrane transport SO NATURE LA English DT Article ID ADP-RIBOSYLATION FACTOR-1; ENDOPLASMIC-RETICULUM; LIVING CELLS; COATOMER BINDING; BREFELDIN-A; G-PROTEIN; EXCHANGE; VESICLES; MUTATION; CLATHRIN AB Cytosolic coat proteins that bind reversibly to membranes have a central function in membrane transport within the secretory pathway(1,2). One well-studied example is COPI or coatomer, a heptameric protein complex that is recruited to membranes by the GTP-binding protein Arf1. Assembly into an electron-dense coat then helps in budding off membrane to be transported between the endoplasmic reticulum (ER) and Golgi apparatus 2. Here we propose and corroborate a simple model for coatomer and Arf1 activity based on results analysing the distribution and lifetime of fluorescently labelled coatomer and Arf1 on Golgi membranes of living cells. We find that activated Arf1 brings coatomer to membranes. However, once associated with membranes, Arf1 and coatomer have different residence times: coatomer remains on membranes after Arf1-GTP has been hydrolysed and dissociated. Rapid membrane binding and dissociation of coatomer and Arf1 occur stochastically, even without vesicle budding. We propose that this continuous activity of coatomer and Arf1 generates kinetically stable membrane domains that are connected to the formation of COPI-containing transport intermediates. This role for Arf1/coatomer might provide a model for investigating the behaviour of other coat protein systems within cells. C1 NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. Rockefeller Univ, Ctr Studies & Phys Biol, New York, NY 10021 USA. BioInformat Serv, Rockville, MD 20854 USA. RP Lippincott-Schwartz, J (reprint author), NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RI Ward, Theresa/E-9650-2013 OI Ward, Theresa/0000-0002-9881-8649 NR 29 TC 175 Z9 175 U1 1 U2 11 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD MAY 9 PY 2002 VL 417 IS 6885 BP 187 EP 193 DI 10.1038/417187a PG 8 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 549RH UT WOS:000175460200046 PM 12000962 ER PT J AU Walsh, TJ Lee, J Dismukes, WE AF Walsh, TJ Lee, J Dismukes, WE TI Decisions about voriconazole versus liposomal amphotericin B SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NCI, Bethesda, MD 20892 USA. Univ Alabama, Birmingham, AL 35294 USA. RP Walsh, TJ (reprint author), NCI, Bethesda, MD 20892 USA. NR 2 TC 16 Z9 17 U1 0 U2 1 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 9 PY 2002 VL 346 IS 19 BP 1499 EP 1499 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 549FE UT WOS:000175433200021 PM 12000825 ER PT J AU Weber, HO Samuel, T Rauch, P Funk, JO AF Weber, HO Samuel, T Rauch, P Funk, JO TI Human p14(ARF)-mediated cell cycle arrest strictly depends on intact p53 signaling pathways SO ONCOGENE LA English DT Article DE INK4a/ARF; p53; p21; 14-3-3 sigma; DNA tumor virus ID P19(ARF) TUMOR-SUPPRESSOR; INK4A LOCUS; P53-DEPENDENT APOPTOSIS; DNA-DAMAGE; MDM2; P14(ARF); CANCER; P21; P16(INK4A); PRODUCT AB The tumor suppressor ARF is transcribed from the INK4a/ARF locus in partly overlapping reading frames with the CDK inhibitor p16(Ink4a). ARF is able to antagonize the MDM2-mediated ubiquitination and degradation of p53, leading to either cell cycle arrest or apoptosis, depending on the cellular context. However, recent data point to additional p53-independent functions of mouse p19(ARF). Little is known about the dependency of human p14(ARF) function on p53 and its downstream genes. Therefore, we analysed the mechanism of p14(ARF)-induced cell cycle arrest in several human cell types. Wild-type HCT116 colon carcinoma cells (p53(+/+)p21(CIP1+/+) 14-3-3sigma(+/+)), but not p53(-/-) counterparts, underwent G(1) and G(2) cell cycle arrest following infection with a p14(ARF)-adenovirus. In p21(CIP1-/-) cells, p14(ARF) did not induce G(1) or G(2) arrest, while 14-3-3sigma(-/-) counterparts were mainly arrested in G(1), pointing to essential roles of p21(CIP1) in G(1) and G(2) arrest and cooperative roles of p21 and 14-3-3sigma in ARF-mediated G(2) arrest. Our data demonstrate a strict p53 and p21(CIP1) dependency of p14(ARF)-induced cell cycle arrest in human cells. C1 Univ Erlangen Nurnberg, Dept Dermatol, Lab Mol Tumor Biol, D-91052 Erlangen, Germany. NCI, Regulat Cell Growth Lab, Frederick, MD 21702 USA. RP Funk, JO (reprint author), Univ Erlangen Nurnberg, Dept Dermatol, Lab Mol Tumor Biol, Hartmannstr 14, D-91052 Erlangen, Germany. NR 42 TC 38 Z9 43 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 9 PY 2002 VL 21 IS 20 BP 3207 EP 3212 DI 10.1038/sj.onc.1205429 PG 6 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 548DP UT WOS:000175373600012 PM 12082636 ER PT J AU Lenox, RH Gould, TD Manji, HK AF Lenox, RH Gould, TD Manji, HK TI Endophenotypes in bipolar disorder SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Review DE biological marker; subclinical trait; vulnerability marker; circadian rhythms; genetics ID GLYCOGEN-SYNTHASE KINASE-3; MANIC-DEPRESSIVE ILLNESS; MAMMALIAN CIRCADIAN CLOCK; CHOLINERGIC REM-INDUCTION; EVENT-RELATED POTENTIALS; SLEEP-PHASE SYNDROME; OPEN-FIELD BEHAVIOR; LONG-QT SYNDROME; INTRACELLULAR CALCIUM; MOOD DISORDERS AB The search for genes in bipolar disorder has provided numerous genetic loci that have been linked to susceptibility to developing the disorder. However, because of the genetic heterogeneity inherent in bipolar disorder, additional strategies may need to be employed to fully dissect the genetic underpinnings. One such strategy involves reducing complex behaviors into their component parts (endophenotypes). Abnormal neurophysiological, biochemical, endocrinological, neuroanatomical, cognitive, and neuropsychological findings are characteristics that often accompany psychiatric illness. It is possible that some of these may eventually be useful in subdefining complex genetic disorders, allowing for improvements in diagnostic assessment, genetic linkage studies, and development of animal models. Findings in patients with bipolar disorder that may eventually be useful as endophenotypes include abnormal regulation of circadian rhythms (the sleep/wake cycle, hormonal rhythms, etc.), response to sleep deprivation, P300 event-related potentials, behavioral responses to psychostimulants and other medications, response to cholinergics, increase in white matter hyperintensities (WHIs), and biochemical observations in peripheral mononuclear cells. Targeting circadian rhythm abnormalities may be a particularly useful strategy because circadian cycles appear to be an inherent evolutionarily conserved function in all organisms and have been implicated in the pathophysiology of bipolar disorder. Furthermore, lithium has been shown to regulate circadian cycles in diverse species, including humans, possibly through inhibition of glycogen synthase kinase 3-beta (GSK-3beta), a known target of lithium. Published 2002 Wiley-Liss, Inc.(dagger) C1 NIMH, Mol Pathophysiol Lab, Bethesda, MD 20892 USA. Aventis Pharmaceut, CNS Res & Drug Discovery, Bridgewater, NJ USA. RP Manji, HK (reprint author), NIMH, Mol Pathophysiol Lab, Bldg 49,Room B1EE16, Bethesda, MD 20892 USA. FU NCRR NIH HHS [RR01 MH59959] NR 239 TC 174 Z9 181 U1 5 U2 16 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 MAY 8 PY 2002 VL 114 IS 4 BP 391 EP 406 DI 10.1002/ajmg.10360 PG 16 WC Genetics & Heredity SC Genetics & Heredity GA 547WM UT WOS:000175355800004 PM 11992561 ER PT J AU Zubenko, GS Hughes, HB Stiffler, JS Zubenko, WN Kaplan, BB AF Zubenko, GS Hughes, HB Stiffler, JS Zubenko, WN Kaplan, BB TI Genome survey for susceptibility loci for recurrent, early-onset major depression: Results at 10cM resolution SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE recurrent, early-onset, major depressive disorder (RE-MDD); mood disorders; genetics; susceptibility genes; sex-specific effects ID ALZHEIMERS-DISEASE; AFFECTIVE-DISORDERS; PSYCHIATRIC-DISORDERS; UNIPOLAR DEPRESSION; SEX-DIFFERENCES; FAMILY HISTORY; PANIC DISORDER; RISK; TWIN; ALLELE AB Recurrent (two or more episodes), early-onset (first episode at less than or equal to25 years) major depressive disorder (RE-MDD) is a strongly familial condition (lambda(first-degree relatives) = 8) whose malignant effects have a significant negative impact on the health and longevity of patients and their family members. The goal of this study was to identify candidate susceptibility loci that influence the development of RE-MDD. We completed a systematic survey of the human genome, conducted at an average resolution of 10 cM, for the identification of simple sequence tandem repeat polymorphisms (SSTRPs) that target susceptibility genes for RE-MDD by virtue of linkage disequilibrium. The efficiency of our association study was enhanced by genotyping pools of DNA from 100 adults with RE-MDD and 100 adult controls who had no personal or family history of mental disorders. Both groups included equal numbers of Caucasian men and women and were matched as closely as possible for age and ethnicity. Allelic associations with RE-MDD were observed for 19 of the 387 SSTRPs in the CHLC Human Screening Set/Weber Version 9. Sixteen of the 19 candidate susceptibility loci revealed significant allelic associations with RE-MDD in men (n = 7) or women (n = 9), but not in both sexes. Evidence for both risk and protective alleles was detected. Two of the candidate susceptibility loci reside within several Mb of loci previously reported-megabases to be linked to "comorbid alcoholism and depression" in families of individuals with alcoholism and to a broadly defined affected phenotype that included recurrent major depression in the families of patients with bipolar disorder. Although it has been suggested that the genes that influence risk for MDD in the two sexes may not be entirely the same, the results of our study suggest that sex specificity of susceptibility loci for RE-MDD may be the rule rather than the exception. The observed preponderance of sex-specific susceptibility loci for RE-MDD suggests that there may be important differences in the molecular pathophysiology of RE-MDD in men and women. Alternatively, our findings may reflect the existence of sex-specific differences in the molecular mechanisms that determine resilience to endogenous or environmental depressogenic stimuli. (C) 2002 Wiley-Liss, Inc. C1 Univ Pittsburgh, Sch Med, Dept Psychiat, Pittsburgh, PA USA. Carnegie Mellon Univ, Dept Biol Sci, Pittsburgh, PA 15213 USA. NIMH, Mol Biol Lab, Bethesda, MD 20892 USA. RP Zubenko, GS (reprint author), WPIC, Room E1230,3811 OHara St, Pittsburgh, PA 15213 USA. FU NIMH NIH HHS [MH48969, MH00540, MH30915, MH60866, MH43261] NR 73 TC 52 Z9 55 U1 2 U2 3 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 MAY 8 PY 2002 VL 114 IS 4 BP 413 EP 422 DI 10.1002/ajmg.10381 PG 10 WC Genetics & Heredity SC Genetics & Heredity GA 547WM UT WOS:000175355800006 PM 11992563 ER PT J AU Shiao, YH Ramakrishna, G Anderson, LM Perantoni, AO Rice, JM Diwan, BA AF Shiao, YH Ramakrishna, G Anderson, LM Perantoni, AO Rice, JM Diwan, BA TI Down-regulation of von Hippel-Lindau protein in N-nitroso compound-induced rat non-clear cell renal tumors SO CANCER LETTERS LA English DT Article DE von Hippel-Lindau; protein down-regulation; N-nitroso compound; non-clear cell; rat kidney tumor ID SUPPRESSOR GENE; KIDNEY TUMORS; HUMAN TISSUES; PRODUCT; VHL; CARCINOGENESIS; METHYLATION; MECHANISMS; EXPRESSION; CARCINOMA AB Non-clear cell rat kidney tumors, inducible by N-nitroso compounds but lacking mutations in the von Hippel-Lindau (VHL) coding sequence, were examined for other VHL alterations. Neither mutations nor DNA methylation was detected in a putative promoter region. By immunohistochemistry, however, VHL protein level was evidently reduced in six of the eight eosinophilic renal epithelial tumors and in all the ten nephroblastomas. Immunoblotting of normal kidney detected two VHL proteins of 20 and 22 kDa in a 16-day-old fetal rat but only 20 kDa protein in an adult rat. This is the first demonstration of VHL alteration at the protein level. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 NCI, Comparat Carcinogenesis Lab, Frederick, MD 21702 USA. Int Agcy Res Canc, Unit Carcinogen Identificat & Evaluat, F-69372 Lyon 08, France. SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. RP Shiao, YH (reprint author), NCI, Comparat Carcinogenesis Lab, Bldg 538,Room 205, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 21 TC 6 Z9 6 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD MAY 8 PY 2002 VL 179 IS 1 BP 33 EP 38 AR PII S0304-3835(01)00863-1 DI 10.1016/S0304-3835(01)00863-1 PG 6 WC Oncology SC Oncology GA 542CW UT WOS:000175026800004 PM 11880179 ER PT J AU Harris, SR Schoeffner, DJ Yoshiji, H Thorgeirsson, UP AF Harris, SR Schoeffner, DJ Yoshiji, H Thorgeirsson, UP TI Tumor growth enhancing effects of vascular endothelial growth factor are associated with increased nitric oxide synthase activity and inhibition of apoptosis in human breast carcinoma xenografts SO CANCER LETTERS LA English DT Article DE breast carcinoma; vascular endothelial growth factor; nitric oxide synthase; proliferation; apoptosis; basement membrane ID PERMEABILITY FACTOR; FACTOR EXPRESSION; BLOOD-FLOW; CELLS; CANCER; ANGIOGENESIS; SURVIVAL AB Previously, we demonstrated the significance of vascular endothelial growth factor (VEGF) in promoting the growth of tetracycline-regulated human VEGF(165) retroviral vector transduced T47-D breast carcinoma cells, particularly at the early stages of tumor development (Cancer Res. 57 (1997) 3924). Here, we showed histologically that the VEGF overexpressing (VEGF (+)) T47-D cells formed a distinct tumor nodule at day 11, while control cells showed no evidence of replication. The VEGF (+) tumors contained large avasculur cavities at days 11 and 21, which were replaced by basement membrane-lined channels at day 30. The number of proliferating tumor cells was not significantly different between the VEGF (+) and control tumors, but the number of apoptotic cells was significantly decreased in the VEGF (+) tumors. Increased nitric oxide synthase (NOS) activity was also observed in the VEGF (+) tumors. These findings indicate that VEGF contributes to tumor growth through inhibition of apoptosis and increased NOS activity, which may be critical during pre-vascular stages of tumor development. Published by Elsevier Science Ireland Ltd. C1 NCI, Tumor Biol & Carcinogenesis Sect, Cellular Carcinogenesis & Tumor Promot Lab, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. RP Thorgeirsson, UP (reprint author), NCI, Tumor Biol & Carcinogenesis Sect, Cellular Carcinogenesis & Tumor Promot Lab, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. NR 20 TC 23 Z9 29 U1 1 U2 2 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 MAY 8 PY 2002 VL 179 IS 1 BP 95 EP 101 AR PII S0304-3835(01)00866-7 DI 10.1016/S0304-3835(01)00866-7 PG 7 WC Oncology SC Oncology GA 542CW UT WOS:000175026800012 PM 11880187 ER PT J AU Malozowski, S Sahlroot, JT AF Malozowski, S Sahlroot, JT TI Sildenafil and physical exertion in men with coronary artery disease SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter C1 NIDDKD, Div Diabet Endocrinol & Metab Dis, Bethesda, MD 20892 USA. US FDA, Ctr Drug Evaluat & Res, Div Biometr 2, Rockville, MD 20857 USA. RP Malozowski, S (reprint author), NIDDKD, Div Diabet Endocrinol & Metab Dis, Bethesda, MD 20892 USA. NR 4 TC 1 Z9 1 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD MAY 8 PY 2002 VL 287 IS 18 BP 2359 EP 2359 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 548PF UT WOS:000175397000014 PM 12001945 ER PT J AU Borio, L Inglesby, T Peters, CJ Schmaljohn, AL Hughes, JM Jahrling, PB Ksiazek, T Johnson, KM Meyerhoff, A O'Toole, T Ascher, MS Bartlett, J Breman, JG Eitzen, EM Hamburg, M Hauer, J Henderson, A Johnson, RT Kwik, G Layton, M Lillibridge, S Nabel, GJ Osterholm, MT Perl, TM Russell, P Tonat, K AF Borio, L Inglesby, T Peters, CJ Schmaljohn, AL Hughes, JM Jahrling, PB Ksiazek, T Johnson, KM Meyerhoff, A O'Toole, T Ascher, MS Bartlett, J Breman, JG Eitzen, EM Hamburg, M Hauer, J Henderson, A Johnson, RT Kwik, G Layton, M Lillibridge, S Nabel, GJ Osterholm, MT Perl, TM Russell, P Tonat, K CA Working Grp Civilian Biodef TI Hemorrhagic fever viruses as biological weapons - Medical and public health management SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Review ID RIFT-VALLEY FEVER; MARCH-APRIL 1972; LASSA FEVER; EBOLA-VIRUS; MARBURG-VIRUS; RHESUS-MONKEYS; WEST AFRICA; JUNIN VIRUS; EXPERIMENTAL-INFECTION; INTRAVENOUS RIBAVIRIN AB Objective To develop consensus-based recommendations for measures to be taken by medical and public health professionals if hemorrhagic fever viruses (HFVs) are used as biological weapons against a civilian population. Participants The Working Group on Civilian Biodefense included 26 representatives from academic medical centers, public health, military services, governmental agencies, and other emergency management institutions. Evidence MEDLINE was searched from January 1966 to January 2002. Retrieved references, relevant material published prior to 1966, and additional sources identified by participants were reviewed. Consensus Process Three formal drafts of the statement that synthesized information obtained in the evidence-gathering process were reviewed by the working group. Each draft incorporated comments and judgments of the members. All members approved the final draft. Conclusions Weapons disseminating a number of HFVs could cause an outbreak of an undifferentiated febrile illness 2 to 21 days later, associated with clinical manifestations that could include rash, hemorrhagic diathesis, and shock. The mode of transmission and clinical course would vary depending on the specific pathogen. Diagnosis may be delayed given clinicians' unfamiliarity with these diseases, heterogeneous clinical presentation within an infected cohort, and lack of widely available diagnostic tests. Initiation of ribavirin therapy in the early phases of illness may be useful in treatment of some of these viruses, although extensive experience is lacking. There are no licensed vaccines to treat the diseases caused by HFVs. C1 Johns Hopkins Sch Med, Johns Hopkins Ctr Civilian Biodef Strategies, Baltimore, MD 21202 USA. Johns Hopkins Sch Publ Hlth, Johns Hopkins Ctr Civilian Biodef Strategies, Baltimore, MD 21202 USA. Johns Hopkins Sch Publ Hlth, Dept Microbiol, Baltimore, MD 21202 USA. Johns Hopkins Sch Med, Dept Microbiol, Baltimore, MD 21202 USA. Johns Hopkins Sch Med, Dept Neurosci, Baltimore, MD 21202 USA. Johns Hopkins Sch Publ Hlth, Dept Neurosci, Baltimore, MD 21202 USA. Johns Hopkins Sch Med, Div Infect Dis, Baltimore, MD USA. Univ Texas, Med Branch, Ctr Biodef, Galveston, TX 77550 USA. NIH, Ctr Clin, Dept Crit Care Med, Bethesda, MD 20892 USA. NIH, Fogarty Int Ctr, Bethesda, MD 20892 USA. NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. USA, Med Res Inst Infect Dis, Frederick, MD USA. Ctr Dis Control & Prevent, Natl Ctr Infect Dis, Atlanta, GA USA. Univ New Mexico, Dept Biol, Albuquerque, NM 87131 USA. Univ New Mexico, Dept Med, Albuquerque, NM 87131 USA. US FDA, Off Commiss, Rockville, MD 20857 USA. US Dept HHS, Off Emergency Preparedness, Rockville, MD USA. US Dept HHS, Off Publ Hlth Preparedness, Washington, DC 20201 USA. Nucl Threat Initiat, Washington, DC USA. New York City Dept Hlth, Bur Communicable Dis, New York, NY 10013 USA. Univ Minnesota, Ctr Infect Dis Res & Policy, Minneapolis, MN USA. RP Borio, L (reprint author), Johns Hopkins Sch Med, Johns Hopkins Ctr Civilian Biodef Strategies, 111 Market Pl,Suite 830, Baltimore, MD 21202 USA. EM Lborio@jhsph.edu NR 140 TC 382 Z9 398 U1 39 U2 234 PU AMER MEDICAL ASSOC PI CHICAGO PA 330 N WABASH AVE, STE 39300, CHICAGO, IL 60611-5885 USA SN 0098-7484 EI 1538-3598 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD MAY 8 PY 2002 VL 287 IS 18 BP 2391 EP 2405 DI 10.1001/jama.287.18.2391 PG 15 WC Medicine, General & Internal SC General & Internal Medicine GA 548PF UT WOS:000175397000028 PM 11988060 ER PT J AU Oh, SH Adler, HJ Raphael, Y Lomax, MI AF Oh, SH Adler, HJ Raphael, Y Lomax, MI TI WDR1 colocalizes with ADF and actin in the normal and noise-damaged chick cochlea SO JOURNAL OF COMPARATIVE NEUROLOGY LA English DT Article DE actin depolymerizing protein; cofilin; cytoskeleton; regeneration; actin dynamics ID HAIR CELL REGENERATION; BASILAR PAPILLA; ACOUSTIC TRAUMA; TECTORIAL MEMBRANE; INTENSE SOUND; INNER-EAR; AUDITORY FUNCTION; CUTICULAR PLATE; MESSENGER-RNA; IN-VIVO AB Auditory hair cells of birds, unlike hair cells in the mammalian organ of Corti, can regenerate following sound-induced loss. We have identified several genes that are upregulated following such an insult. One gene, WDR1, encodes the vertebrate homologue of actin-interacting protein 1, which interacts with actin depolymerization factor (ADF) to enhance the rate of actin filament cleavage. We examined WDR1 expression in the developing, mature, and noise-damaged chick cochlea by in situ hybridization and immunocytochemistry. In the mature cochlea, WDR1 mRNA was detected in hair cells, homogene cells, and cuboidal cells, all of which contain high levels of F-actin. In the developing inner ear, WDR1 mRNA was detected in homogene cells and cuboidal cells by embryonic day 7, in the undifferentiated sensory epithelium by day 9, and in hair cells at embryonic day 16. We also demonstrated colocalization of WDR1, ADF, and F-actin in all three cell types in the normal and noise-damaged cochlea. Immediately after acoustic overstimulation, WDR1 mRNA was seen in supporting cells. These cells contribute to the structural integrity of the basilar papilla, the maintenance of the ionic barrier at the reticular lamina, and the generation of new hair cells. These results indicate that one of the immediate responses of the supporting cell after noise exposure is to induce WDR1 gene expression and thus to increase the rate of actin filament turnover. These results suggest that WDR1 may play a role either in restoring cytoskeletal integrity in supporting cells or in a cell signaling pathway required for regeneration. (C) 2002 Wiley-Liss, Inc. C1 Univ Michigan, Sch Med, Kresge Hearing Res Inst, Dept Otolaryngol Head & Neck Surg, Ann Arbor, MI 48109 USA. Seoul Natl Univ, Coll Med, Dept Otolaryngol Head & Neck Surg, Seoul 110744, South Korea. NIDCD, NIH, Bethesda, MD 20892 USA. RP Lomax, MI (reprint author), Univ Michigan, Sch Med, Kresge Hearing Res Inst, Dept Otolaryngol Head & Neck Surg, 9301E MSRBIII Box 0648, Ann Arbor, MI 48109 USA. RI Oh, Seung Ha/J-5540-2012 FU NIDCD NIH HHS [DC 02492, DC 01634] NR 66 TC 17 Z9 19 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 0021-9967 J9 J COMP NEUROL JI J. Comp. Neurol. PD MAY 8 PY 2002 VL 448 IS 4 BP 399 EP 409 DI 10.1002/cne.10265 PG 11 WC Neurosciences; Zoology SC Neurosciences & Neurology; Zoology GA 558QW UT WOS:000175978900007 PM 12115702 ER PT J AU Singh, P Jones, LA Foley, CK White, PS Pedersen, LG AF Singh, P Jones, LA Foley, CK White, PS Pedersen, LG TI Structural aspects of phencyclidines: crystal structure and quantum mechanical calculations for 1-[1-(2-thienyl)cyclohexyl] piperidine and its hydrochloride SO JOURNAL OF MOLECULAR STRUCTURE LA English DT Article DE phencyclidines; 1-[1-(2-thienyl)cyclohexyl] piperidine; TCP-HCl; ab initio; X-ray AB Crystal structure determinations for neutral and protonated phencyclidine, 1-[1-(2-thienyl)cyclohexyl] piperidine, show that the piperidine-axial conformer on the cyclohexane ring is present in the neutral compound, and that the piperidine-equatorial conformer in the protonated species. Quantum mechanical calculations in vacuo using ab initio techniques also arrive at the same conclusion. A search of the crystallographic literature via the Cambridge structural database reveals that all protonated phencyclidines assume the piperidine-equatorial conformation, and most neutral phencyclidines assume the piperidine-axial conformation. (C) 2002 Elsevier Science B.V. All rights reserved. C1 N Carolina Cent Univ, Durham, NC 27707 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. N Carolina State Univ, Dept Chem, Raleigh, NC 27685 USA. Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. RP Pedersen, LG (reprint author), Univ N Carolina, Dept Chem, CB 3290, Chapel Hill, NC 27599 USA. RI Pedersen, Lee/E-3405-2013; OI Pedersen, Lee/0000-0003-1262-9861; Foley, Charles/0000-0001-6578-9629 NR 21 TC 1 Z9 1 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-2860 J9 J MOL STRUCT JI J. Mol. Struct. PD MAY 8 PY 2002 VL 608 IS 1 BP 55 EP 62 AR PII S0022-2860(01)00932-2 DI 10.1016/S0022-2860(01)00932-2 PG 8 WC Chemistry, Physical SC Chemistry GA 535DP UT WOS:000174630200008 ER PT J AU Golinelli, MP Hughes, SH AF Golinelli, MP Hughes, SH TI Nontemplated nucleotide addition by HIV-1 reverse transcriptase SO BIOCHEMISTRY LA English DT Article ID STEADY-STATE KINETICS; DNA-POLYMERASE; NONNUCLEOSIDE INHIBITORS; STRAND TRANSFER; NUCLEOCAPSID PROTEIN; SECONDARY STRUCTURE; CRYSTAL-STRUCTURE; MECHANISM; SUBSTRATE; COMPLEX AB We studied the kinetics of nontemplated nucleotide addition by the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) using model substrates derived front the 3' end of HIV-1 minus-strand strong-stop DNA. The addition of a nontemplated nucleotide was highly dependent on the nature of the base (fastest addition with dATP), type of nucleoside, and pH of the reaction buffer. The salt concentration, presence or absence of nucleocapsid protein, and nature of the blunt-ended duplex (DNA/DNA versus RNA/DNA) had only limited effects. The efficiency and base specificity were strongly affected by the sequence at the 3' end of the blunt-ended duplex. In every case, nontemplated nucleotide addition was much slower than templated polymerization. The K-d for the incoming dNTP with an RT bound to a blunt-ended duplex was at least 1000-fold higher than with a duplex with a template overhang. At concentrations normally found in vivo, ATP can compete with dNTPs for binding to the polymerase active site and reduce the efficiency of nontemplated nucleotide addition. Although a stable ternary complex RT/DNA/dNTP could be readily detected by gel retardation assays if the DNA had a template overhang. stable ternary complexes were not observed with a blunt-ended duplex substrate. At in vivo concentrations of dNTPs (5-10 muM), nontemplated nucleotide addition occurred, but it was very inefficient and the rate of nontemplated polymerization is at least 10000-fold slower than the rate of templated polymerization. We could conclude that, in vivo, the unfavorable binding of the incoming dNTP, low concentration of dNTPs, the presence of a large concentration of ATP, and the inability to form a stable ternary complex prior to the polymerization step collaborate to reduce the efficiency of nontemplated nucleotide addition. C1 NCI, HIV Drug Resistance Program, Frederick, MD 21702 USA. RP Hughes, SH (reprint author), NCI, HIV Drug Resistance Program, POB B,Bldg 539,Room 130A, Frederick, MD 21702 USA. RI Golinelli, Marie-Pierre/K-4287-2013 OI Golinelli, Marie-Pierre/0000-0002-6738-8631 NR 50 TC 18 Z9 18 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD MAY 7 PY 2002 VL 41 IS 18 BP 5894 EP 5906 DI 10.1021/bi0160415 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 548AJ UT WOS:000175365300021 PM 11980493 ER PT J AU Baumgarten, G Knuefermann, P Kalra, D Gao, F Taffet, GE Michael, L Blackshear, PJ Carballo, E Sivasubramanian, N Mann, DL AF Baumgarten, G Knuefermann, P Kalra, D Gao, F Taffet, GE Michael, L Blackshear, PJ Carballo, E Sivasubramanian, N Mann, DL TI Load-dependent and -independent regulation of proinflammatory cytokine and cytokine receptor gene expression in the adult mammalian heart SO CIRCULATION LA English DT Article DE hypertrophy; gene expression; growth factors; cytokines; remodeling ID NECROSIS-FACTOR-ALPHA; FELINE MYOCARDIUM; PROTEIN EXPRESSION; TRANSGENIC MICE; PATHWAY; FAILURE AB Background-Although previous studies have examined the effects of acute hemodynamic pressure overload on proinflammatory cytokine gene expression, the effects of sustained hemodynamic overloading have not been examined. Methods and Results-Sustained hemodynamic pressure overloading was produced in mice by transverse constriction of the aorta. Proinflammatory cytokine and cytokine receptor gene expression were determined by ribonuclease protection assays (RPA) at 6 hours and at 3, 7, 14 and 35 days after banding. M-mode echocardiography was used to assess left ventricular structure and function at identical time points. RPA showed that tumor necrosis factor (TNF), interleukin (IL)-1beta, and IL-6 mRNA levels were maximal at 6 hours and returned to baseline levels within 72 hours. There was a significant increase in IL-1RII and IL-6Ralpha receptor mRNA levels after overloading but no significant increase in TNFR1, TNFR2, IL-1RI, or gp130 mRNA levels. The transient increase in expression of proinflammatory cytokine gene expression was not explained by changes in left ventricular loading conditions, left ventricular wall stress, desensitization of proinflammatory genes, or decreased nuclear factor-kappabeta activation. It is interesting that transverse constriction of the aorta provoked an increase in the expression of tristetraprolin, a homeostatic zinc finger protein that is known to destabilize TNF mRNA. Conclusion-Sustained hemodynamic overloading provokes a transient increase in proinflammatory cytokine and cytokine receptor gene expression; however, the decrease in proinflammatory cytokine gene expression occurred in the absence of changes in loading conditions, suggesting that the expression of proinflammatory cytokines in the heart is regulated, at least in part, by load-dependent and load-independent mechanisms. C1 Houston VAMC, Dept Med, Winters Ctr Heart Failure Res, Houston, TX 77030 USA. Baylor Coll Med, DeBakey Heart Ctr, Houston, TX 77030 USA. Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. RP Mann, DL (reprint author), Houston VAMC, Dept Med, Winters Ctr Heart Failure Res, MS 524,6565 Fannin, Houston, TX 77030 USA. OI Mann, Douglas /0000-0002-2516-0145 FU NHLBI NIH HHS [HL-42250-10/10, P50 HL-O6H, R01 HL58081-01, R01 HL61543-01] NR 14 TC 83 Z9 83 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 MAY 7 PY 2002 VL 105 IS 18 BP 2192 EP 2197 DI 10.1161/01.CIR.0000015608.37608.18 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 550KK UT WOS:000175502000014 PM 11994254 ER PT J AU Zou, MF Agoston, GE Belov, Y Kopajtic, T Katz, JL Newman, AH AF Zou, MF Agoston, GE Belov, Y Kopajtic, T Katz, JL Newman, AH TI Enantioselective synthesis of S-(+)-2 beta-carboalkoxy-3 alpha-[bis(4-fluorophenyl)methoxy]tropanes as novel probes for the dopamine transporter SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID N-SUBSTITUTED 3-ALPHA-TROPANE; UPTAKE INHIBITORS; COCAINE ABUSE; 3-ALPHA-(DIPHENYLMETHOXY)TROPANE ANALOGS; BENZTROPINE ANALOGS; RHESUS-MONKEYS; BINDING; LIGANDS; PHARMACOTHERAPIES; DISCOVERY AB Synthesis of a series of pure S-(+)-2beta-carboalkoxy-3alpha-[bis(4-fluotophenyl)metlioxy]tropanes (>99% ee) was achieved by employing a chiral amine-induced asymmetric reaction of tropinone with methyl cyanoformate as the key step. In this series, all of the S-(+)-enantiomers were 2-fold more potent than their racemic mixtures and all displayed high-affinity binding for DAT (K-i - 13-40 nM). These data support previous findings of significant divergence in structural requirements for high-affinity DAT binding among tropane-based inhibitors. Furthermore, the 2-substituent in the 3alpha-[bis(4-fluorophenyl)methoxy]tropane series is well tolerated at the DAT but not at SERT (K-i = 690-2040 nM), or muscarinic M-1 receptors (K-i - 133-4380 nM) resulting in highly selective DAT ligands that may provide new leads toward a cocaine-abuse therapeutic. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Natl Inst Drug Abuse Intramural Res Program, Med Chem Sect, NIH, Baltimore, MD 21224 USA. ChromBA, State Coll, PA 16803 USA. Natl Inst Drug Abuse Intramural Res Program, Psychobiol Sect, NIH, Baltimore, MD 21224 USA. RP Newman, AH (reprint author), Natl Inst Drug Abuse Intramural Res Program, Med Chem Sect, NIH, Baltimore, MD 21224 USA. NR 27 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 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD MAY 6 PY 2002 VL 12 IS 9 BP 1249 EP 1252 AR PII S0960-894X(02)00155-5 DI 10.1016/S0960-894X(02)00155-5 PG 4 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 550LG UT WOS:000175504100004 PM 11965364 ER PT J AU Houlard, M Arudchandran, R Regnier-Ricard, F Germani, A Gisselbrecht, S Blank, U Rivera, J Varin-Blank, N AF Houlard, M Arudchandran, R Regnier-Ricard, F Germani, A Gisselbrecht, S Blank, U Rivera, J Varin-Blank, N TI Vav1 is a component of transcriptionally active complexes SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE Vav; nuclear translocation; nuclear factor of activated T cells; calcium influx; protein subdomains ID T-CELL RECEPTOR; FOCAL CONTACT PROTEIN; MAST-CELLS; TYROSINE PHOSPHORYLATION; PROTOONCOGENE PRODUCT; SIGNAL-TRANSDUCTION; MEMBRANE DOMAINS; GENE-EXPRESSION; ACTIVATION; BINDING AB The importance of the hematopoietic protooncogene Vav1 in immune cell function is widely recognized although its regulatory mechanisms are not completely understood. Here, we examined whether Vav1 has a nuclear function, as past studies have reported its nuclear localization. Our findings provide a definitive demonstration of Vav1 nuclear localization in a receptor stimulation-dependent manner and reveal a critical role for the COOH-terminal Src homology 3 (SH3) domain and a nuclear localization sequence within the pleckstrin homology domain. Analysis of DNA-bound transcription factor complexes revealed nuclear Vav1 as an integral component of transcriptionally active nuclear factor of activated T cells (NFAT)- and nuclear factor (NF)kappaB-like complexes, and the COOH-terminal SH3 domain as being critical in their formation. Thus, we describe a novel nuclear role for Vav1 as a component and facilitator of NFAT and NFkappaB-like transcriptional activity. C1 Hop Cochin, INSERM, Inst Cochin Genet Mol, Unite Oncol Cellulaire & Mol 363, F-75014 Paris, France. NIAMSD, Mol Inflammat Sect, NIH, Bethesda, MD 20892 USA. Inst Pasteur, Unite Immunoallergie, F-75015 Paris, France. RP Varin-Blank, N (reprint author), Hop Cochin, INSERM, Inst Cochin Genet Mol, Unite Oncol Cellulaire & Mol 363, 27 Rue Faubourg St Jacques, F-75014 Paris, France. NR 52 TC 55 Z9 57 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 MAY 6 PY 2002 VL 195 IS 9 BP 1115 EP 1127 DI 10.1084/jem.20011701 PG 13 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 560ZF UT WOS:000176110700004 PM 11994417 ER PT J AU Mascola, JR AF Mascola, JR TI Passive transfer studies to elucidate the role of antibody-mediated protection against HIV-1 SO VACCINE LA English DT Article; Proceedings Paper CT Sympopsium on Global HIV Therapeutics - HIV Vaccines CY JUN 07-09, 2001 CL KAROLINSKA INST, STOCKHOLM, SWEDEN HO KAROLINSKA INST DE passive transfer studies; antibody-mediated protections; HIV-1 ID IMMUNODEFICIENCY-VIRUS TYPE-1; NEUTRALIZING MONOCLONAL-ANTIBODIES; VAGINAL TRANSMISSION; CHIMERIC VIRUS; INFECTION; IMMUNIZATION; MACAQUES; PREVENTION; CHALLENGE; ESCAPE AB In order to understand immune correlates of protection and to develop effective immunization strategies, it is important to know if antibodies can confer protection against HIV-1 infection or disease. The recent development of the pathogenic simian/human immunodeficiency virus (SHIV)-macaque model based on env genes from primary HIV-1 isolates permits the in vivo evaluation of anti-HIV-1 envelope glycoprotein immune responses. Using this model, we and others initially showed that passively infused antibody can protect against an intravenous SHIV-challenge. However. HIV-1 is most often transmitted across mucosal surfaces and the intravenous challenge model may not accurately predict the role of antibody in protection against mucosal exposure. We, therefore, adapted the SHIV89.6PD model to allow evaluation of anti-HIV-1 antibodies against vaginal challenge. In order to make comparisons to our prior intravenous challenge study, we used the same SHIV89.6PD stock and antibodies. Our data show that antibodies can confer protection against vaginal exposure to a pathogenic SHIV; if virus transmission occurs, their presence can ameliorate the subsequent pathogenic manifestations of virus infection. Compared to our previous intravenous challenge study, greater protection was achieved after vaginal challenge. Because the highest level of protection occurred when the most potent combinations of antibodies were used, the data confirm that in vitro neutralization assays on peripheral blood mononuclear cells (PBMC) targets cells are a relevant measure of protective antibody activity. Published by Elsevier Science Ltd. C1 NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. RP Mascola, JR (reprint author), NIAID, Vaccine Res Ctr, NIH, 40 Convent Dr, Bethesda, MD 20892 USA. NR 17 TC 76 Z9 79 U1 0 U2 0 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 MAY 6 PY 2002 VL 20 IS 15 SI SI BP 1922 EP 1925 AR PII S0264-410X(02)00068-3 DI 10.1016/S0264-410X(02)00068-3 PG 4 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 552MR UT WOS:000175624600008 PM 11983246 ER PT J AU Nabel, GJ AF Nabel, GJ TI HIV vaccine strategies SO VACCINE LA English DT Article; Proceedings Paper CT Sympopsium on Global HIV Therapeutics - HIV Vaccines CY JUN 07-09, 2001 CL KAROLINSKA INST, STOCKHOLM, SWEDEN HO KAROLINSKA INST DE HIV; vaccine; immunity ID VIVO GENE DELIVERY; IMMUNODEFICIENCY-VIRUS; IN-VIVO; DNA IMMUNIZATION; VECTORS; PROTECTION; MALARIA; IMMUNOGENICITY; EXPRESSION; PARTICLES AB Traditional methods of vaccine development have not produced effective vaccines for several prevalent infectious diseases, including AIDS, malaria and tuberculosis. These difficult diseases call attention to the importance of new approaches that profit from modem technologies. Successful efforts in the past have typically taken advantage of naturally occurring, protective immune responses, but this avenue is not readily available in certain cases, such as in HIV infection, where the immune system rarely confers protective immunity. However, there are alternative strategies and areas of research that may facilitate the development of highly effective vaccines. These include the identification of immunogens that elicit broadly neutralizing antibodies, determination of the molecular and cellular basis for immune responses to the components of the infectious agent, the identification of relevant forms of viral proteins for antigen presentation, stimulation of relevant T-cell types. and enhancement of antigen-presenting, dendritic cell function. Answering these basic research questions will aid in rational vaccine design. It is also extremely important to optimize techniques for the testing and production of new vaccines including the quantitation of immune responses in animals and in humans, identification of surrogate markers of immune protection, streamlined vaccine production, and rapid evaluation of candidate vaccines for testing in clinical trials. We have put these ideas into practice in two recent studies in which we generated enhanced cytotoxic T lymphocyte (CTL) responses, while retaining robust humoral responses, to wild-type viral proteins by immunizing mice with genetically modified forms of HIV-1 Env, Gag and Pol delivered in the form of plasmid DNA expression vectors. Published by Elsevier Science Ltd. C1 NIAID, NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. RP Nabel, GJ (reprint author), NIAID, NIH, Vaccine Res Ctr, 40 Convent Dr, Bethesda, MD 20892 USA. NR 31 TC 56 Z9 59 U1 1 U2 1 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 MAY 6 PY 2002 VL 20 IS 15 SI SI BP 1945 EP 1947 AR PII S0264-410X(02)00074-9 DI 10.1016/S0264-410X(02)00074-9 PG 3 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 552MR UT WOS:000175624600014 PM 11983251 ER PT J AU Amara, RR Villinger, F Altman, JD Lydy, SL O'Neil, SP Staprans, SI Montefiori, DC Xu, Y Herndon, JG Wyatt, LS Candido, MA Kozyr, NL Earl, PL Smith, JM Ma, HL Grimm, BD Hulsey, ML McClure, HM McNicholl, JM Moss, B Robinson, HL AF Amara, RR Villinger, F Altman, JD Lydy, SL O'Neil, SP Staprans, SI Montefiori, DC Xu, Y Herndon, JG Wyatt, LS Candido, MA Kozyr, NL Earl, PL Smith, JM Ma, HL Grimm, BD Hulsey, ML McClure, HM McNicholl, JM Moss, B Robinson, HL TI Control of a mucosal challenge and prevention of AIDS by a multiprotein DNA/MVA vaccine SO VACCINE LA English DT Article; Proceedings Paper CT Symposium on Global HIV Therapeutics - HIV Vaccines CY JUN 07-09, 2001 CL KAROLINSKA INST, STOCKHOLM, SWEDEN HO KAROLINSKA INST DE mucosal challenged; multiprotein; heterologous prime; boost ID HUMAN-IMMUNODEFICIENCY-VIRUS; RHESUS-MONKEYS; NEUTRALIZING ANTIBODIES; PATHOGENIC SIV; INFECTION; RESPONSES; TYPE-1 AB Heterologous prime/boost regimens have the potential for raising high levels of immune responses. Here, we report that DNA priming followed by a recombinant modified vaccinia Ankara (rMVA) booster has controlled a highly pathogenic immunodeficiency virus challenge in a Rhesus macaque model. Both the DNA and rMVA components of the vaccine expressed multiple immunodeficiency virus proteins. Two DNA inoculations at 0 and 8 weeks and a single rMVA booster at 24 weeks effectively controlled an intrarectal challenge administered 7 months after the booster. These highly promising findings provide hope that a relatively simple multiprotein DNA/MVA vaccine can help to control the AIDS epidemic. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Emory Univ, Vaccine Res Ctr, Yerkes Reg Primate Res Ctr, Atlanta, GA 30329 USA. Emory Univ, Sch Med, Vaccine Res Ctr, Dept Pathol & Lab Med, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Vaccine Res Ctr, Dept Microbiol & Immunol, Atlanta, GA 30322 USA. Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA. NIAID, NIH, Viral Dis Lab, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Vaccine Res Ctr, Div Infect Dis,Dept Med, Atlanta, GA 30322 USA. Natl Ctr Infect Dis, Div AIDS STD & TB Lab Res, Ctr Dis Control & Prevent, Atlanta, GA 30330 USA. RP Robinson, HL (reprint author), Emory Univ, Vaccine Res Ctr, Yerkes Reg Primate Res Ctr, 954 Gatewood Rd NE, Atlanta, GA 30329 USA. EM hrobins@rmy.emory.edu FU NCRR NIH HHS [P51 RR 00165]; NIAID NIH HHS [P01 AI 43045]; NIDA NIH HHS [P30 DA 12121] NR 14 TC 64 Z9 72 U1 1 U2 3 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 MAY 6 PY 2002 VL 20 IS 15 SI SI BP 1949 EP 1955 AR PII S0264-410X(02)00076-2 DI 10.1016/S0264-410X(02)00076-2 PG 7 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 552MR UT WOS:000175624600015 PM 11983252 ER PT J AU Muthumani, K Kudchodkar, S Zhang, DH Bagarazzi, ML Kim, JJ Boyer, JD Ayyavoo, V Pavlakis, GN Weiner, DB AF Muthumani, K Kudchodkar, S Zhang, DH Bagarazzi, ML Kim, JJ Boyer, JD Ayyavoo, V Pavlakis, GN Weiner, DB TI Issues for improving multiplasmid DNA vaccines for HIV-1 SO VACCINE LA English DT Article; Proceedings Paper CT Sympopsium on Global HIV Therapeutics - HIV Vaccines CY JUN 07-09, 2001 CL KAROLINSKA INST, STOCKHOLM, SWEDEN HO KAROLINSKA INST DE DNA vaccine; HIV-1; cytokines immune responses ID MEDIATED PROTECTIVE IMMUNITY; HERPES-SIMPLEX VIRUS-2; RHESUS-MONKEYS; VACCINATION; EXPRESSION; RESPONSES; GAG; PLASMID; IMMUNOGENICITY; SEQUENCE AB Since the first reports of plasmid vaccines. there have been substantial changes made to the design of plasmid backbones. as well as to the antibiotic resistance markers chosen for clinical vectors compared with first generation vectors. These changes aid manufacturing, production and scale up and at the same time aid conceptual safety by limiting the ability of the vaccines to transfer useful genetic selection genes to other bacterial infectious agents. In contrast, there has been little change to the original promoters or polyadenlyation tracts in the last decade. We have learned that these first generation plasmid vaccines for HIV-1 appear very well tolerated in humans. However, while safe and immunogenic, improving the immune potency of DNA vaccines is a critical goal for this technology. The combination of antigens used should be carefully examined for possible immune interference. Such interference may only become apparent when each component of the vaccine is tested individually. This interference also suggests one mechanism of immune pathogenesis possibly by HIV-1. Optimization of the immune response can come through manipulation of the transfection efficiency, expression or through the use of various T cell and B cell plasmid adjuvants. It is likely that the combination of such advancements will significantly improve the clinical phenotype of this important vaccine modality. (C) 2002 Elsevier Science Ltd. All rights reserved. C1 Univ Penn, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA. NCI, Frederick, MD 21701 USA. Univ Pittsburgh, Dept Microbiol, Pittsburgh, PA USA. Viral Genomix, Philadelphia, PA USA. St Christophers Hosp Children, Dept Pediat, Philadelphia, PA 19133 USA. RP Weiner, DB (reprint author), Univ Penn, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA. RI Muthumani, Karuppiah/D-1092-2009; Weiner, David/H-8579-2014; OI Ayyavoo, Velpandi/0000-0002-9043-0885 NR 26 TC 28 Z9 28 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 MAY 6 PY 2002 VL 20 IS 15 SI SI BP 1999 EP 2003 AR PII S0264-410X(02)00086-5 DI 10.1016/S0264-410X(02)00086-5 PG 5 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 552MR UT WOS:000175624600025 PM 11983262 ER PT J AU Nemukhin, AV Topol, IA Burt, SK AF Nemukhin, AV Topol, IA Burt, SK TI Energy profiles for the rate-limiting stage of the serine protease prototype reaction SO INTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY LA English DT Article DE serine protease; reaction path; quantum chemistry modeling ID FRAGMENT POTENTIAL METHOD AB The energy profiles of reaction, modeling the rate-limiting stage of serine protease catalyzed transformations, have been computed by quantum chemistry, methods. The model includes fragments of the residues of the catalytic triad (serine, histidine, and aspartic acid), two water molecules as an oxyanion hole, represented by effective fragment potentials, and a formamide molecule as a substrate. Geometry optimizations have been performed along the reaction coordinate, chosen as the distance between oxygen of serine and carbon of substrate, by using the Hartree-Fock method. The density functional theory B3LYP/6-31+G(d,p) calculations have been employed to recalculate energies along the reaction path in the gas phase and in the dielectric environment. The computed barrier heights are fairly consistent with the data cited in the literature. (C) 2002 Wiley Periodicals, Inc. C1 NCI, SAIC Frederick, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. Moscow MV Lomonosov State Univ, Dept Chem, Moscow 119899, Russia. RP Burt, SK (reprint author), NCI, SAIC Frederick, Adv Biomed Comp Ctr, POB B, Frederick, MD 21702 USA. RI Nemukhin, Alexander/P-9662-2015 NR 13 TC 13 Z9 13 U1 0 U2 0 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0020-7608 J9 INT J QUANTUM CHEM JI Int. J. Quantum Chem. PD MAY 5 PY 2002 VL 88 IS 1 BP 34 EP 40 DI 10.1002/qua.10076 PG 7 WC Chemistry, Physical; Mathematics, Interdisciplinary Applications; Physics, Atomic, Molecular & Chemical SC Chemistry; Mathematics; Physics GA 543GR UT WOS:000175093500004 ER PT J AU Brenner, RA AF Brenner, RA TI Childhood drowning is a global concern - Prevention needs a multifaceted approach SO BRITISH MEDICAL JOURNAL LA English DT Editorial Material ID CHILDREN C1 NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. RP Brenner, RA (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. NR 13 TC 21 Z9 24 U1 0 U2 1 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0959-535X J9 BRIT MED J JI Br. Med. J. PD MAY 4 PY 2002 VL 324 IS 7345 BP 1049 EP 1050 DI 10.1136/bmj.324.7345.1049 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 549YY UT WOS:000175476500003 PM 11991894 ER PT J AU Toth, A Kedei, N Szabo, T Wang, Y Blumberg, PM AF Toth, A Kedei, N Szabo, T Wang, Y Blumberg, PM TI Thapsigargin binds to and inhibits the cloned vanilloid receptor-1 SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE vanilloid receptor subtype 1; thapsigargin; capsaicin; resiniferatoxin binding; VR1-transfected CHO cells; sarcoplasmatic reticulum ATPase ID ACTIVATED CALCIUM CHANNELS; CAPSAICIN RECEPTOR; NEURONS; RESINIFERATOXIN; CAPSAZEPINE; CELLS AB We investigated the effect of thapsigargin, a well-known sarcoplasmic reticulum ATPase (SERCA) inhibitor, on the non-specific Ca2+ channel vanilloid receptor-1 (VR1) in CHO-VR1 cells. We found that thapsigargin inhibited the VR-1 mediated Ca-45(2+) uptake of CHO-VR1 cells (IC50 = 6.4 +/- 1.9 muM) and the (3)[H]RTX binding to VR1 (IC50 = 4.0 +/- 1.3 muM). Further analysis revealed that thapsigargin is a mixed-type inhibitor, suggesting both direct and indirect interactions between thapsigargin and the capsaicin binding site of VR1. Thapsigargin alone transiently elevated the [Ca2+](i) in CHO-VR1 cells (EC50 = 44 nM). However, Ca-45(2+) uptake was not detected after thapsigargin treatment, indicating that the emptying of the thapsigargin sensitive intracellular pools of Ca2+ was responsible for the elevated [Ca2+](i) level rather than the activation of VR-1. We conclude that thapsigargin represents a new prototype of a VR1 inhibitor and that caution should be exercised in interpreting the effects of thapsigargin, especially when it is used in the micromolar range to inhibit SERCA activity. (C) 2002 Elsevier Science (USA). All rights reserved. C1 NCI, Mol Mechanisms Tumor Promot Sect, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. RP Blumberg, PM (reprint author), NCI, Mol Mechanisms Tumor Promot Sect, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bldg 37,Room 3A01,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. RI Toth, Attila/F-4859-2010 OI Toth, Attila/0000-0001-6503-3653 NR 21 TC 25 Z9 27 U1 1 U2 2 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD MAY 3 PY 2002 VL 293 IS 2 BP 777 EP 782 AR PII S0006-291X(02)00293-0 DI 10.1016/S0006-291X(02)00293-0 PG 6 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 550DP UT WOS:000175488600020 PM 12054538 ER PT J AU Ponamarev, MV Longley, MJ Nguyen, D Kunkel, TA Copeland, WC AF Ponamarev, MV Longley, MJ Nguyen, D Kunkel, TA Copeland, WC TI Active site mutation in DNA polymerase gamma associated with progressive external ophthalmoplegia causes error-prone DNA synthesis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID P55 ACCESSORY SUBUNIT; MITOCHONDRIAL-DNA; MULTIPLE DELETIONS; ESCHERICHIA-COLI; KLENOW FRAGMENT; FIDELITY; REPLICATION; MTDNA; IDENTIFICATION; DISORDER AB Progressive external ophthalmoplegia (PEO) is a heritable mitochondrial disorder characterized by the accumulation of multiple point mutations and large deletions in mtDNA. Autosomal dominant PEO was recently shown to co-segregate with a heterozygous Y955C mutation in the human gene encoding the sole mitochondrial DNA polymerase, DNA polymerase gamma (pol gamma). Since Tyr-955 is a highly conserved residue critical for nucleotide recognition among family A DNA polymerases, we analyzed the effects of the Y955C mutation on the kinetics and fidelity of DNA synthesis by the purified human mutant polymerase in complex with its accessory subunit. The Y955C enzyme retains a wild-type catalytic rate (k(cat)) but suffers a 45-fold decrease in apparent binding affinity for the incoming nucleoside triphosphate (K-m). The Y955C derivative is 2-fold less accurate for base pair substitutions than wild-type pol gamma despite the action of intrinsic exonucleolytic proofreading. The fall mutator effect of the Y955C substitution was revealed by genetic inactivation of the exonuclease, and error rates for certain mismatches were elevated by 10-100-fold. The error-prone DNA synthesis observed for the Y955C pol gamma is consistent with the accumulation of mtDNA mutations in patients with PEO. C1 NIEHS, Genet Mol Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Copeland, WC (reprint author), NIEHS, Genet Mol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 29 TC 91 Z9 94 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 MAY 3 PY 2002 VL 277 IS 18 BP 15225 EP 15228 DI 10.1074/jbc.C200100200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 550PA UT WOS:000175510400002 PM 11897778 ER PT J AU Song, J Kwon, O Chen, SL Daruwala, R Eck, P Park, JB Levine, M AF Song, J Kwon, O Chen, SL Daruwala, R Eck, P Park, JB Levine, M TI Flavonoid inhibition of sodium-dependent vitamin C transporter 1 (SVCT1) and glucose transporter isoform 2 (GLUT2), intestinal transporters for vitamin C and glucose SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BRUSH-BORDER MEMBRANE; L-ASCORBIC-ACID; DEHYDROASCORBIC ACID; HUMAN-NEUTROPHILS; DIABETES-MELLITUS; DNA-DAMAGE; QUERCETIN; ABSORPTION; CELLS; FAMILY AB Vitamin C and flavonoids, polyphenols with uncertain function, are abundant in fruits and vegetables. We postulated that flavonoids have a novel regulatory action of delaying or inhibiting absorption of vitamin C and glucose, which are structurally similar. From six structural classes of flavonoids, at least 12 compounds were chosen for studies. We investigated the effects of selected flavonoids on the intestinal vitamin C transporter SVCT1(h) by transfecting and overexpressing SVCT1(h) in Chinese hamster ovary cells. Flavonoids reversibly inhibited vitamin C transport in transfected cells with IC50 values of 10-50 muM, concentrations expected to have physiologic consequences. The most potent inhibitor class was flavonols, of which quercetin is most abundant in foods. Because Chinese hamster ovary cells have endogenous vitamin C transport, we expressed SVCT1(h) in Xenopus laevis oocytes to study the mechanism of transport inhibition. Quercetin was a reversible and non-competitive inhibitor of ascorbate transport; K-i 17.8 muM. Quercetin was a potent non-competitive inhibitor of GLUT2 expressed in Xenopus oocytes; K-i 22.8 muM. When diabetic rats were administered glucose with quercetin, hyperglycemia was significantly decreased compared with administration of glucose alone. Quercetin also significantly decreased ascorbate absorption in normal rats given ascorbate plus quercetin compared with rats given ascorbate alone. Quercetin was a specific transport inhibitor, because it did not inhibit intestinal sugar transporters GLUT5 and SGLT1 that were injected and expressed in Xenopus oocytes. Quercetin inhibited but was not transported by SVCT1(h). Considered together, these data show that flavonoids modulate vitamin C and glucose transport by their respective intestinal transporters and suggest a new function for flavonoids. C1 NIDDK, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. USDA, Phytonutrients Lab, Beltsville Human Nutr Res Ctr, Beltsville, MD 20705 USA. RP Levine, M (reprint author), NIDDK, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bldg 10,Rm 4D52,MSC 1372, Bethesda, MD 20892 USA. RI Chen, Shenglin/B-4049-2010; OI Eck, Peter/0000-0003-2371-9774 NR 58 TC 114 Z9 119 U1 1 U2 26 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 MAY 3 PY 2002 VL 277 IS 18 BP 15252 EP 15260 DI 10.1074/jbc.M110496200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 550PA UT WOS:000175510400007 PM 11834736 ER PT J AU Heron-Milhavet, L LeRoith, D AF Heron-Milhavet, L LeRoith, D TI Insulin-like growth factor I induces MDM2-dependent degradation of p53 via the p38 MAPK pathway in response to DNA damage SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NUCLEOTIDE EXCISION-REPAIR; NUCLEAR EXCLUSION; SIGNALING PATHWAY; HUMAN FIBROBLASTS; UV-RADIATION; MDM2; RECEPTOR; CELLS; PHOSPHORYLATION; ERCC-1 AB In many tissues, the insulin-like growth factor I (IGF-I) receptor (IGF-IR) is known to functionally oppose apoptosis. Recently, we demonstrated a direct role for the IGF-IR in the rescue of DNA-damaged fibroblasts by activating a DNA repair pathway (Heron-Milhavet, L., Karas, M., Goldsmith, C. M., Baum, B. J., and LeRoith, D. (2001) J. Biol. Chem. 276, 18185-18192). p53 is a nuclear transcription factor that can block progression of the cell cycle, modulate DNA repair, and trigger apoptosis. In this work, we tested the effect of IGF-I on the regulation of the p53 signaling cascade. The DNA-damaging agent 4-nitroquinoline 1-oxide was applied to NIH-3T3 cells overexpressing normal IGF-IRs (NWTb3 cells). We showed that after 4-nitroquinoline 1-oxide-induced DNA damage, IGF-I induced exclusion of the p53 protein from the nucleus and led to its degradation in the cytoplasm, whereas p53 mRNA was unaffected. Degradation of the p53 protein was associated with an increase in MDM2, an upstream modulator of the half-life and activity of the p53 protein. p53 degradation was also associated with down-regulation of p21. We further showed that the effects of IGF-I on mdm2 transcription and on MDM2/p19 ARF association were mediated by the p38 MAPK pathway. In conclusion, we describe a novel role for IGF-I in the regulation of the MDM2/p53/p21 signaling pathway during DNA damage. C1 NIDDK, Sect Cellular & Mol Physiol, Clin Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP LeRoith, D (reprint author), NIDDK, Sect Cellular & Mol Physiol, Clin Endocrinol Branch, NIH, Bldg 10,Rm 8D12, Bethesda, MD 20892 USA. NR 34 TC 72 Z9 73 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 3 PY 2002 VL 277 IS 18 BP 15600 EP 15606 DI 10.1074/jbc.M111142200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 550PA UT WOS:000175510400052 PM 11877395 ER PT J AU O'Neill, T Giarratani, L Chen, P Iyer, L Lee, CH Bobiak, M Kanai, F Zhou, BB Chung, JH Rathbun, GA AF O'Neill, T Giarratani, L Chen, P Iyer, L Lee, CH Bobiak, M Kanai, F Zhou, BB Chung, JH Rathbun, GA TI Determination of substrate motifs for human Chk1 and hCds1/Chk2 by the oriented peptide library approach SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DNA-DAMAGE CHECKPOINT; ATAXIA-TELANGIECTASIA GENE; CELL-CYCLE REGULATION; XENOPUS EGG EXTRACTS; CDK FAMILY MEMBER; PROTEIN-KINASE; S-PHASE; SCHIZOSACCHAROMYCES-POMBE; IONIZING-RADIATION; CRYSTAL-STRUCTURE AB Mammalian Chk1 and Chk2 are two Ser/Thr effector kinases that play critical roles in DNA damage-activated cell cycle checkpoint signaling pathways downstream of ataxia telangiectasia-mutated and ataxia telangiectasia-related. Endogenous substrates have been identified for human hCds1/Chk2 and Chk1; however, the sequences surrounding the substrate residues appear unrelated, and consensus substrate motifs for the two Ser/Thr kinases remain unknown. We have utilized peptide library analyses to develop specific, highly preferred substrate motifs for hCds1/Chk2 and Chk1. The optimal motifs are similar for both kinases and most closely resemble the previously identified Chk1 and hCds1/Chk2 substrate target sequences in Cdc25C and Cdc25A, the regulation of which plays an important role in S and G(2)M arrest. Essential residues required for the definition of the optimal motifs were also identified. Utilization of the peptides to assay the substrate specificities and catalytic activities of Chk1 and hCds1/Chk2 revealed substantial differences between the two Ser/Thr kinases. Structural modeling analyses of the peptides into the Chk1 catalytic cleft were consistent with Chk1 kinase assays defining substrate suitability. The library-derived substrate preferences were applied in a genome-wide search program, revealing novel targets that might serve as substrates for hCds1/Chk2 or Chk1 kinase activity. C1 Harvard Univ, Childrens Hosp, Sch Med, Dept Pediat,Ctr Blood Res, Boston, MA 02115 USA. Agouron Pharmaceut Inc, San Diego, CA 92121 USA. Harvard Univ, Sch Med, Ctr Res Comp, Boston, MA 02115 USA. NIH, HLBI, Lab Biochem Genet, Bethesda, MD 20892 USA. GlaxoSmithKline, Dept Oncol Res, King Of Prussia, PA 19406 USA. Harvard Univ, Beth Israel Deaconess Hosp, Sch Med, Dept Med, Boston, MA 02115 USA. RP Rathbun, GA (reprint author), Harvard Univ, Childrens Hosp, Sch Med, Dept Pediat,Ctr Blood Res, 200 Longwood Ave,Warren Alpert Bldg,Rm 135, Boston, MA 02115 USA. OI Iyer, Lakshmanan/0000-0001-9167-6396 NR 91 TC 106 Z9 108 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD MAY 3 PY 2002 VL 277 IS 18 BP 16102 EP 16115 DI 10.1074/jbc.M111705200 PG 14 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 550PA UT WOS:000175510400118 PM 11821419 ER PT J AU Urbaneja, MA Wu, M Casas-Finet, JR Karpel, RL AF Urbaneja, MA Wu, M Casas-Finet, JR Karpel, RL TI HIV-1 nucleocapsid protein as a nucleic acid chaperone: Spectroscopic study of its helix-destabilizing properties, structural binding specificity, and annealing activity SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE nucleic acid chaperone; nucleocapsid protein; HIV-1 hybridization; annealing ID IMMUNODEFICIENCY-VIRUS TYPE-1; RNA PACKAGING SIGNAL; STEM-LOOP SL2; REVERSE TRANSCRIPTION; RECOGNITION ELEMENT; GENOME RECOGNITION; STRAND TRANSFER; NMR STRUCTURE; DNA; EQUILIBRIA AB Assembly of infectious retroviral particles involves recognition of specific sequences on the viral RNA by the nucleocapsid (NC) domain of the Gag polyprotein, and subsequent stoichiometric binding of the processed NC protein along the entire length of the RNA. NC proteins also act as nucleic acid chaperones. They accelerate nucleic acid hybridization and strand exchange, which may be critical during the initial stages of reverse transcription. In order to better understand these properties, we have studied the nucleic acid helix-destabilizing t(m)-depressing) and binding activities of HIV-1 NCp7 protein with a variety of substrates, and the real-time kinetics of NC-induced strand exchange. At low ionic strength (0.01 M Na phosphate, pH 7.0) and saturating levels of protein, NCp7 displays moderate helix-destabilizing activity on double-stranded DNA. Saturating levels of NCp7 lowered the t(m) of a synthetic 28 base-pair 28(+)/28(-) oligonucleotide duplex by about 10 deg. C (51 to 41 degreesC). The presence of single-stranded calf thymus DNA (equimolar with duplex) eliminated the t(m) depression, whereas double-stranded calf thymus DNA only altered the t(m) of the 28-mer duplex by about 2 deg. C. Similar effects were seen with duplexes with single-stranded overhangs or internal single-stranded gaps. Binding experiments utilizing intrinsic tryptophan quenching indicated significant affinity (K-d about 0.1 muM) for both single-stranded and double-stranded forms of the 28-mer in 0.01 M sodium phosphate at 25 degreesC, although long-chain (calf thymus double-stranded) DNA displayed a much lower affinity. The effects of NCp7 on the kinetics of nucleic acid annealing, strand exchange, and strand displacement were determined by use of oligonucleotides with end-labeled fluorophores serving as donor-acceptor pairs. NCp7 accelerated all these reactions. In the strand exchange reaction, an imperfect duplex, 28(+)/21(-), was reacted with a perfect complement, 28(-). The kinetics of 28(+)/28(-) annealing in this reaction did not conform to a simple bimolecular model, but could be well fit to the sum of two exponential decays. Addition of stoichiometric levels of NCp7 increased the rate constants of both components, and significantly increased the fraction of exchange associated with the rapid process. Increasing levels of 28(-) also increased the rapid fraction, as well as the rapid rate constant. This concentration dependence indicates that, although the kinetic decays appear biexponential, at least one of the steps is bimolecular. Simple annealing reactions, 28(+) with 28(-), could be fit to single-exponential decays, and their magnitudes in the presence of NCp7 were comparable to the rapid step of annealing observed for exchange reactions, suggesting that this step is connected with annealing. Strand dissociation during exchange was monitored by placing the fluorescent acceptor on the 21(-) strand. The results, though complex, suggest that the slow step of exchange is largely associated with the dissociation of the shorter oligonucleotide. Analogous experiments were performed with variants of these oligonucleotides, and the results are in line with the 28(+)/21(-)/28(-) experiments. On the basis of an analysis of the effect of increasing levels of 28(-) on the formation of the perfect 28 bp duplex from the imperfect duplex, we propose that NCp7 forms a ternary complex intermediate with imperfect duplex and 28(-), and suggest several ways by which such an intermediate would facilitate strand exchange. (C) 2002 Elsevier Science Ltd. All rights reserved C1 NCI, AIDS Vaccine Program, SAIC Frederick, Frederick, MD 21702 USA. Univ Maryland Baltimore Cty, Dept Chem & Biochem, Baltimore, MD 21250 USA. RP Urbaneja, MA (reprint author), Univ Basque Country, Dept Bioquim & Biol Mol, CSIC, E-48080 Bilbao, Spain. EM gbpurarm@lg.ehu.es; karpel@umbc.edu FU NCI NIH HHS [N01-CO-56000]; NIGMS NIH HHS [GM-52049] NR 34 TC 82 Z9 82 U1 2 U2 4 PU ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD MAY 3 PY 2002 VL 318 IS 3 BP 749 EP 764 DI 10.1016/S0022-2836(02)00043-8 PG 16 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 554ZR UT WOS:000175767700012 PM 12054820 ER PT J AU Chen, X Howard, OMZ Yang, XY Wang, LH Oppenheim, JJ Krakauer, T AF Chen, X Howard, OMZ Yang, XY Wang, LH Oppenheim, JJ Krakauer, T TI Effects of Shuanghuanglian and Qingkailing, two multi-components of traditional Chinese medicinal preparations, on human leukocyte function SO LIFE SCIENCES LA English DT Article DE Qingkailing (QKL); Shuanghuanglian (SHHL); cytokine; chemokine; chemotaxis NF-kappa B ID NF-KAPPA-B; TOXIC SHOCK SYNDROME; T-CELL ACTIVATION; HERBAL MEDICINES; ANTIINFLAMMATORY ACTIVITY; STAPHYLOCOCCUS-AUREUS; TRANSCRIPTION FACTORS; CYTOKINE PRODUCTION; HUMAN-DISEASE; CHEMOKINES AB Qingkailing (QKL) and Shuanghuanglian (SHHL) are two commonly used Chinese herbal preparations with reported antiinflammatory activity. The effects of these two preparations on the capacity of staphylococcal toxic shock syndrome toxin 1 (TSST-1) to stimulate the production of cytokines (IL-1beta, IL-6, TNF-alpha, IFN-gamma) and chemokines (MIP-1alpha, MIP-1beta and MCP-1) by peripheral blood mononuclear cell (PBMC) was tested. We also evaluated their effect on LPS-stimulated NF-kappaB transcriptional activity in a THP-1 cell line, and on human monocyte chemotactic response to chemoattractants. Non-cytotoxic concentrations of QKL (0.1similar to2%) and SHHL (6similar to120 mug) significantly inhibited production of cytokines and chemokines in a dose-dependent manner (P < 0.05). Both, QKL at 1:100 and SHHL at 60 mug/ml, markedly inhibited RANTES, MIP-1alpha, SDF-1alpha and fmLP induced human monocyte migration (P < 0.05 or 0.01). QKL (1%) did not inhibit monocyte chemotaxis induced by super-or sub-optimal concentrations of fMLP (10(-5), 10(-6) and 10(-10) M), but only inhibited chemotaxis induced by optimal concentrations of fMLP at 10(-7), 10(-8) and 10(-9) M. QKL (0.1% or 1%) and SHHL(6 or 60 mug/ml) markedly inhibited LPS-induced NF-kappaB activity in THP-1 cells. The results suggested that the pharmacological basis for the antiinflammatory effects of QKL and SHHL is the result of suppression of NF-kappaB regulated gene transcription, leading to suppressed production of proinflammatory cytokine and chemokine. Interference with leukocyte chemotaxis also contributes to the anti inflammatory and immunomodulating effects of these medicinals. Identification of the responsible components in these two herbal preparations may yield compounds suitable for structural modification into potent novel drugs. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NCI, Ctr Canc Res, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NCI, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. USA, Med Res Inst Infect Dis, Dept Immunol & Mol Biol, Frederick, MD 21702 USA. RP Chen, X (reprint author), NCI, Ctr Canc Res, Mol Immunoregulat Lab, Bldg 560,Rm 31-19, Frederick, MD 21702 USA. RI Howard, O M Zack/B-6117-2012; Chen, Xin/I-6601-2015 OI Howard, O M Zack/0000-0002-0505-7052; Chen, Xin/0000-0002-2628-4027 FU NCCIH NIH HHS [Y2-AT-9002]; PHS HHS [N01-C0-12400] NR 67 TC 31 Z9 32 U1 1 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0024-3205 J9 LIFE SCI JI Life Sci. PD MAY 3 PY 2002 VL 70 IS 24 BP 2897 EP 2913 AR PII S0024-3205(02)01541-2 DI 10.1016/S0024-3205(02)01541-2 PG 17 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 565MV UT WOS:000176374800005 PM 12269401 ER PT J AU Kapranov, P Cawley, SE Drenkow, J Bekiranov, S Strausberg, RL Fodor, SPA Gingeras, TR AF Kapranov, P Cawley, SE Drenkow, J Bekiranov, S Strausberg, RL Fodor, SPA Gingeras, TR TI Large-scale transcriptional activity in chromosomes 21 and 22 SO SCIENCE LA English DT Article ID DNA-SEQUENCE; GENES; RNAS AB The sequences of the human chromosomes 21 and 22 indicate that there are approximately 770 well-characterized and predicted genes. In this study, empirically derived maps identifying active areas of RNA transcription on these chromosomes have been constructed with the use of cytosolic polyadenylated RNA obtained from 11 human cell lines. Oligonucleotide arrays containing probes spaced on average every 35 base pairs along these chromosomes were used. When compared with the sequence annotations available for these chromosomes, it is noted that as much as an order of magnitude more of the genomic sequence is transcribed than accounted for by the predicted and characterized exons. C1 Affymetrix, Santa Clara, CA 95051 USA. NCI, Bethesda, MD 20892 USA. RP Gingeras, TR (reprint author), Affymetrix, Santa Clara, CA 95051 USA. OI Gingeras, Thomas/0000-0001-9106-3573 NR 14 TC 564 Z9 588 U1 2 U2 21 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 MAY 3 PY 2002 VL 296 IS 5569 BP 916 EP 919 DI 10.1126/science.1068597 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 549KF UT WOS:000175442500046 PM 11988577 ER PT J AU Celeste, A Petersen, S Romanienko, PJ Fernandez-Capetillo, O Chen, HT Sedelnikova, OA Reina-San-Martin, B Coppola, V Meffre, E Difilippantonio, MJ Redon, C Pilch, DR Olaru, A Eckhaus, M Camerini-Otero, RD Tessarollo, L Livak, F Manova, K Bonner, WM Nussenzweig, MC Nussenzweig, A AF Celeste, A Petersen, S Romanienko, PJ Fernandez-Capetillo, O Chen, HT Sedelnikova, OA Reina-San-Martin, B Coppola, V Meffre, E Difilippantonio, MJ Redon, C Pilch, DR Olaru, A Eckhaus, M Camerini-Otero, RD Tessarollo, L Livak, F Manova, K Bonner, WM Nussenzweig, MC Nussenzweig, A TI Genomic instability in mice lacking histone H2AX SO SCIENCE LA English DT Article ID DOUBLE-STRAND BREAKS; END-JOINING PATHWAY; ATM-DEFICIENT MICE; DNA-REPAIR; ATAXIA-TELANGIECTASIA; TARGETED DISRUPTION; IONIZING-RADIATION; VERTEBRATE CELLS; RECOMBINATION; STABILITY AB Higher order chromatin structure presents a barrier to the recognition and repair of DNA damage. Double-strand breaks (DSBs) induce histone H2AX phosphorylation, which is associated with the recruitment of repair factors to damaged DNA. To help clarify the physiological role of H2AX, we targeted H2AX in mice. Although H2AX is not essential for irradiation-induced cell-cycle checkpoints, H2AX(-/-) mice were radiation sensitive, growth retarded, and immune deficient, and mutant males were infertile. These pleiotropic phenotypes were associated with chromosomal instability, repair defects, and impaired recruitment of Nbs1, 53bp1, and Brca1, but not Rad51, to irradiation-induced foci. Thus, H2AX is critical for facilitating the assembly of specific DNA-repair complexes on damaged DNA. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. NIH, Mol Pharmacol Lab, Bethesda, MD 20892 USA. Rockefeller Univ, Howard Hughes Med Inst, Lab Mol Immunol, New York, NY 10021 USA. NIH, Mouse Canc Genet Program, Frederick, MD 20892 USA. NCI, Dept Genet, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Dept Microbiol & Immunol, Baltimore, MD 21201 USA. NIH, Vet Resources Program, Natl Ctr Res Resources, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, Mol Cytol Core Facil, New York, NY 10021 USA. Mem Sloan Kettering Canc Ctr, Program Mol Biol, New York, NY 10021 USA. RP Nussenzweig, A (reprint author), NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RI Coppola, Vincenzo/E-2917-2011; Fernandez-Capetillo, Oscar/H-3508-2015; Reina-San-Martin, Bernardo/I-9484-2016 OI Coppola, Vincenzo/0000-0001-6163-1779; Fernandez-Capetillo, Oscar/0000-0002-2690-6885; Reina-San-Martin, Bernardo/0000-0003-2083-6166 FU Intramural NIH HHS [Z99 CA999999] NR 33 TC 896 Z9 927 U1 9 U2 45 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 MAY 3 PY 2002 VL 296 IS 5569 BP 922 EP 927 DI 10.1126/science.1069398 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 549KF UT WOS:000175442500048 PM 11934988 ER PT J AU Douek, DC Brenchley, JM Betts, MR Ambrozak, DR Hill, BJ Okamoto, Y Casazza, JP Kuruppu, J Kuntsman, K Wolinsky, S Grossman, Z Dybul, M Oxenius, A Price, DA Connors, M Koup, RA AF Douek, DC Brenchley, JM Betts, MR Ambrozak, DR Hill, BJ Okamoto, Y Casazza, JP Kuruppu, J Kuntsman, K Wolinsky, S Grossman, Z Dybul, M Oxenius, A Price, DA Connors, M Koup, RA TI HIV preferentially infects HIV-specific CD4(+) T cells SO NATURE LA English DT Article ID ACTIVE ANTIRETROVIRAL THERAPY; ANTIGENIC-STIMULATION; IMMUNE-RESPONSES; LYMPHOID-TISSUE; IMMUNODEFICIENCY; REPLICATION; CD8(+); DISCONTINUATION; IDENTIFICATION; INDIVIDUALS AB HIV infection is associated with the progressive loss of CD4(+) T cells through their destruction or decreased production(1,2). A central, yet unresolved issue of HIV disease is the mechanism for this loss, and in particular whether HIV-specific CD4(+) T cells are preferentially affected(3-5). Here we show that HIV-specific memory CD4(+) T cells in infected individuals contain more HIV viral DNA than other memory CD4(+) T cells, at all stages of HIV disease. Additionally, following viral rebound during interruption of antiretroviral therapy, the frequency of HIV viral DNA in the HIV-specific pool of memory CD4(+) T cells increases to a greater extent than in memory CD4(+) T cells of other specificities. These findings show that HIV-specific CD4(+) T cells are preferentially infected by HIV in vivo. This provides a potential mechanism to explain the loss of HIV-specific CD4(+) T-cell responses, and consequently the loss of immunological control of HIV replication(6). Furthermore, the phenomenon of HIV specifically infecting the very cells that respond to it adds a cautionary note to the practice of structured therapy interruption. C1 NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NCI, Dept Expt Transplantat & Immunol, Med Branch, NIH, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Med, Dallas, TX 75390 USA. Northwestern Univ, Sch Med, Dept Infect Dis, Chicago, IL 60611 USA. Tel Aviv Univ, Dept Physiol & Pharmacol, IL-69978 Tel Aviv, Israel. NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. John Radcliffe Hosp, Nuffield Dept Clin Med, Oxford OX3 9DU, England. RP Douek, DC (reprint author), NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. RI Grossman, Zvi/A-9643-2008; Wolinsky, Steven/B-2893-2012; Price, David/C-7876-2013; Oxenius, Annette/G-7794-2015 OI Price, David/0000-0001-9416-2737; NR 32 TC 777 Z9 799 U1 3 U2 35 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD MAY 2 PY 2002 VL 417 IS 6884 BP 95 EP 98 DI 10.1038/417095a PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 546ZM UT WOS:000175307200045 PM 11986671 ER PT J AU Christian, MC Goldberg, JL Killen, J Abrams, JS McCabe, MS Mauer, JK Wittes, RE AF Christian, MC Goldberg, JL Killen, J Abrams, JS McCabe, MS Mauer, JK Wittes, RE TI Sounding board - A central institutional review board for multi-institutional trials. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material C1 NCI, Bethesda, MD 20892 USA. NIAID, Bethesda, MD 20892 USA. RP Christian, MC (reprint author), NCI, Bethesda, MD 20892 USA. NR 18 TC 76 Z9 78 U1 0 U2 1 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 02 PY 2002 VL 346 IS 18 BP 1405 EP 1408 DI 10.1056/NEJM200205023461814 PG 4 WC Medicine, General & Internal SC General & Internal Medicine GA 546MZ UT WOS:000175279000013 PM 11986418 ER PT J AU Ando, Y Figg, WD AF Ando, Y Figg, WD TI Irinotecan in small-cell lung cancer SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NCI, Bethesda, MD 20892 USA. RP Ando, Y (reprint author), NCI, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 3 TC 4 Z9 4 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 02 PY 2002 VL 346 IS 18 BP 1414 EP 1414 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 546MZ UT WOS:000175279000024 PM 11986422 ER PT J AU Hull, KM Kastner, DL Balow, JE AF Hull, KM Kastner, DL Balow, JE TI Hereditary periodic fever SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter ID FAMILIAL MEDITERRANEAN FEVER C1 NIAMSD, Bethesda, MD 20892 USA. NIDDKD, Bethesda, MD 20892 USA. RP Hull, KM (reprint author), NIAMSD, Bethesda, MD 20892 USA. NR 6 TC 12 Z9 13 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 02 PY 2002 VL 346 IS 18 BP 1415 EP 1415 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 546MZ UT WOS:000175279000026 PM 11986423 ER PT J AU Kelley, RI Takada, K Aksentijevich, I AF Kelley, RI Takada, K Aksentijevich, I TI Hereditary periodic fever SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 Kennedy Inst, Baltimore, MD 21205 USA. NIAMSD, Bethesda, MD 20892 USA. RP Kelley, RI (reprint author), Kennedy Inst, Baltimore, MD 21205 USA. NR 2 TC 0 Z9 0 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC/NEJM PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD MAY 02 PY 2002 VL 346 IS 18 BP 1416 EP 1416 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 546MZ UT WOS:000175279000028 ER PT J AU Caballero, OL Resto, V Patturajan, M Meerzaman, D Guo, MZ Engles, J Yochem, R Ratovitski, E Sidransky, D Jen, J AF Caballero, OL Resto, V Patturajan, M Meerzaman, D Guo, MZ Engles, J Yochem, R Ratovitski, E Sidransky, D Jen, J TI Interaction and colocalization of PGP9.5 with JAB1 and p27(Kip1) SO ONCOGENE LA English DT Article DE PGP9.5/UCH-L1; JAB1; p27(Kip1); lung cancer; nuclear localization; yeast two-hybrid system ID CELL LUNG-CANCER; DEUBIQUITINATING ENZYMES; PROTEIN COMPLEX; CDK INHIBITOR; UBIQUITIN; DEGRADATION; PROTEASOME; P27; YEAST; CYCLE AB PGP9.5 (UCH-L1) is a member of the ubiquitin C-terminal hydrolase (UCH) family of proteins that is expressed in neuronal tissues. Our previous studies have shown that PGP9.5 was highly expressed in primary lung cancers and lung cancer cell lines. Additionally, the frequency of PGP9.5 over expression increases with tumor stage, indicating that PGP9.5 may play a role in lung cancer tumorigenesis. We used the yeast two-hybrid system to identify proteins that interact with PGP9.5. We show that PGP9.5 interacts with at least three proteins, one of which is JAB1, a Jun activation domain binding protein that can bind to p27(Kip1) and is involved in the cytoplasmic transportation of p27(Kip1) for its degradation. We also show that PGP9.5 is associated with JAB1 in vitro and in vivo; and that both proteins can be a part of a heteromeric complex containing p27(Kip1) in the nucleus in lung cancer cells. Furthermore, under serum-restimulation, nuclear translocation of both PGP9.5 and JAB1 coincides with a reduced level of P27(Kip1) in the nucleus. In contrast, when cells are contact inhibited, both PGP9.5 and JAB1 became more perinuclear and cytoplasmic in localization while p27(Kip1) was present only in the nucleus. Therefore, PGP9.5 may contribute to p27(Kip1) degradation via its interaction and nuclear translocation with JAB1. C1 NCI, Lab Populat Genet, Ctr Canc Res, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Otolaryngol Head & Neck Surg, Div Head & Neck Canc Res, Baltimore, MD 21205 USA. RP Jen, J (reprint author), NCI, Lab Populat Genet, Ctr Canc Res, Bldg 41,Rm D702,41 Lib Dr, Bethesda, MD 20892 USA. RI Resto, Vicente/D-9891-2014 FU NCI NIH HHS [CA 58184] NR 33 TC 81 Z9 87 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD MAY 2 PY 2002 VL 21 IS 19 BP 3003 EP 3010 DI 10.1038/sj/onc/1205390 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 546FM UT WOS:000175262700007 PM 12082530 ER PT J AU Summers, RM AF Summers, RM TI Challenges for computer-aided diagnosis for CT colonography SO ABDOMINAL IMAGING LA English DT Article DE computed tomography, colon; computed tomography, three-dimensional reconstruction; colon cancer; image processing ID CURVED CROSS-SECTIONS; VIRTUAL COLONOSCOPY; COLON; SEGMENTATION; OPTIMIZATION; FEASIBILITY; ENDOSCOPY AB Computer-aided diagnosis for computed tomographic colonography is in its infancy but has the potential to improve sensitivity and decrease costs for colonic polyp detection. This article reviews the current state of research in this nascent field and explores major challenges and avenues for future work. C1 NIH, Dept Diagnost Radiol, Bethesda, MD 20892 USA. RP Summers, RM (reprint author), NIH, Dept Diagnost Radiol, Bldg 10,Room 1C660,10 Ctr Dr,MSC 1182, Bethesda, MD 20892 USA. NR 26 TC 44 Z9 47 U1 0 U2 0 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0942-8925 EI 1432-0509 J9 ABDOM IMAGING JI Abdom. Imaging PD MAY-JUN PY 2002 VL 27 IS 3 BP 268 EP 274 DI 10.1007/s00261-001-0168-7 PG 7 WC Gastroenterology & Hepatology; Radiology, Nuclear Medicine & Medical Imaging SC Gastroenterology & Hepatology; Radiology, Nuclear Medicine & Medical Imaging GA 544TD UT WOS:000175176000007 PM 12173357 ER PT J AU Bryant, LH Jordan, EK Bulte, JWM Herynek, V Frank, JA AF Bryant, LH Jordan, EK Bulte, JWM Herynek, V Frank, JA TI Pharmacokinetics of a high-generation dendrimer-Gd-DOTA SO ACADEMIC RADIOLOGY LA English DT Article; Proceedings Paper CT Contrast Media Research Meeting (CMR99) CY SEP 12-17, 1999 CL WOODSTOCK, VERMONT ID IMAGING CONTRAST AGENTS; MONOCLONAL-ANTIBODY; RELAXOMETRY; RELAXATION; IONS C1 Warren Grant Magnuson Clin Ctr, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. NINCDS, Neuroimaging Branch, NIH, Bethesda, MD 20892 USA. RP Bryant, LH (reprint author), Warren Grant Magnuson Clin Ctr, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. RI Bulte, Jeff/A-3240-2008 OI Bulte, Jeff/0000-0003-1202-1610 NR 17 TC 8 Z9 8 U1 0 U2 2 PU ASSOC UNIV RADIOLOGISTS PI OAK BROOK PA 820 JORIE BLVD, OAK BROOK, IL 60523-2251 USA SN 1076-6332 J9 ACAD RADIOL JI Acad. Radiol. PD MAY PY 2002 VL 9 SU 1 BP S29 EP S33 DI 10.1016/S1076-6332(03)80390-2 PG 5 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 546VR UT WOS:000175296700007 PM 12019888 ER PT J AU Bulte, JWM Zhang, SC van Gelderen, P Herynek, V Jordan, EK Janssen, CHC Duncan, ID Frank, JA AF Bulte, JWM Zhang, SC van Gelderen, P Herynek, V Jordan, EK Janssen, CHC Duncan, ID Frank, JA TI Magnetically labeled glial cells as cellular MR contrast agents SO ACADEMIC RADIOLOGY LA English DT Article; Proceedings Paper CT Contrast Media Research Meeting (CMR99) CY SEP 12-17, 1999 CL WOODSTOCK, VERMONT ID BRAIN; TRANSPLANTATION; RECEPTOR; LINE C1 Univ Wisconsin, Sch Vet Med, Dept Med Sci, Madison, WI 53706 USA. Natl Inst Neurol Disorders & Stroke, Neuroimaging Branch, NIH, Bethesda, MD USA. NIH, In Vivo NMR Ctr, Bethesda, MD 20892 USA. NIH, Ctr Clin, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. RP Bulte, JWM (reprint author), Johns Hopkins Univ, Sch Med, Dept Radiol, 217 Traylor Bldg,720 Rutland Ave, Baltimore, MD 21205 USA. RI Bulte, Jeff/A-3240-2008 OI Bulte, Jeff/0000-0003-1202-1610 NR 9 TC 12 Z9 13 U1 0 U2 0 PU ASSOC UNIV RADIOLOGISTS PI OAK BROOK PA 820 JORIE BLVD, OAK BROOK, IL 60523-2251 USA SN 1076-6332 J9 ACAD RADIOL JI Acad. Radiol. PD MAY PY 2002 VL 9 SU 1 BP S148 EP S150 DI 10.1016/S1076-6332(03)80424-5 PG 3 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 546VR UT WOS:000175296700041 PM 12019854 ER PT J AU Damadzic, R Shuangshoti, S Giblen, G Herman, MM AF Damadzic, R Shuangshoti, S Giblen, G Herman, MM TI Neuritic pathology is lacking in the entorhinal cortex, subiculum and hippocampus in middle-aged adults with schizophrenia, bipolar disorder or unipolar depression SO ACTA NEUROPATHOLOGICA LA English DT Article DE neuritic pathology; entorhinal cortex; schizophrenia; bipolar disease; unipolar depression ID ALZHEIMERS-DISEASE; COGNITIVE IMPAIRMENT; ELDERLY SCHIZOPHRENICS; VOLUME REDUCTION; NEURONAL DENSITY; PSYCHOSIS; DEMENTIA; PLAQUES; TANGLES; BRAINS AB Earlier reports have provided conflicting results regarding the association between Alzheimer's disease (AD) and mental disorders. Using a well-characterized postmortem series of 40 middle-aged human brains, we have performed quantitative analysis of neurofibrillary tangles and senile plaques in the entorhinal cortex, subiculum and rostral hippocampus in 9 subjects with schizophrenia, 8 with bipolar disorder, 12 with depression, and 11 age- and sex-matched controls. No significant differences were found among the four groups. Our study indicates that the Alzheimer-type changes, which might be related to the likelihood of AD development later in life, are not increased in middle-aged subjects with mental illness. The result also supports the more recent reports that have demonstrated no increased incidence of AD in mentally ill patients. C1 NIMH, Sect Neuropathol, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Herman, MM (reprint author), NIMH, Sect Neuropathol, Clin Brain Disorders Branch, NIH, Bldg 36,Room 3A24,36 Convent Dr, Bethesda, MD 20892 USA. NR 29 TC 16 Z9 16 U1 1 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0001-6322 J9 ACTA NEUROPATHOL JI Acta Neuropathol. PD MAY PY 2002 VL 103 IS 5 BP 488 EP 494 DI 10.1007/s00401-001-0496-2 PG 7 WC Clinical Neurology; Neurosciences; Pathology SC Neurosciences & Neurology; Pathology GA 542NF UT WOS:000175049500010 PM 11935265 ER PT J AU Kaplan, RC Heckbert, SR Koepsell, TD Rosendaal, FR Furberg, CD Cooper, LS Psaty, BM AF Kaplan, RC Heckbert, SR Koepsell, TD Rosendaal, FR Furberg, CD Cooper, LS Psaty, BM CA Cardiovascular Hlth Study Investig TI Calcium channel blocker use and gastrointestinal tract bleeding among older adults SO AGE AND AGEING LA English DT Letter ID RISK C1 Albert Einstein Coll Med, Dept Epidemiol & Social Med, Bronx, NY 10461 USA. Cardiovasc Hlth Res Unit, Seattle, WA 98101 USA. Leiden Univ, Med Ctr, Dept Clin Epidemiol, Leiden, Netherlands. Wake Forest Univ, Bowman Gray Sch Med, Dept Publ Hlth Sci, Winston Salem, NC 27157 USA. NHLBI, Div Epidemiol & Clin Applicat, Prevent Studies Res Grp, Bethesda, MD 20892 USA. RP Kaplan, RC (reprint author), Albert Einstein Coll Med, Dept Epidemiol & Social Med, Belfer Bldg Room 1308C, Bronx, NY 10461 USA. RI Kaplan, Robert/A-2526-2011 NR 9 TC 4 Z9 4 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0002-0729 J9 AGE AGEING JI Age Ageing PD MAY PY 2002 VL 31 IS 3 BP 217 EP U1 DI 10.1093/ageing/31.3.217 PG 2 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 556JX UT WOS:000175846600013 PM 12006312 ER PT J AU Purohit, V Russo, D AF Purohit, V Russo, D TI Cellular and molecular mechanisms of alcoholic hepatitis: introduction and summary of the symposium SO ALCOHOL LA English DT Editorial Material DE alcoholic hepatitis; Kupffer cells; leukocytes; chemokines; adhesion molecules ID ETHANOL; LIVER; DIET AB The National Institute on Alcohol Abuse and Alcoholism and the Office of Rare Diseases, National Institutes of Health, sponsored a satellite symposium on "Cellular and Molecular Mechanisms of Alcoholic Hepatitis" at the 24th Annual Scientific Meeting of the Research Society on Alcoholism, Montreal, Quebec, Canada, June 2001. Alcohol intake is a major cause of hepatitis that may lead to alcoholic cirrhosis-a major cause of death in the United States. In up to one third of heavy drinkers alcoholic hepatitis develops, which is characterized by liver cell death and infiltration of leukocytes in hepatic parenchyma. Although leukocytes have been implicated in the pathogenesis of alcoholic hepatitis, the underlying cellular and molecular mechanisms by which leukocytes migrate to hepatic parenchyma and initiate tissue injury are not clear. For this symposium, 10 speakers were invited to address the following aspects of the mechanisms of alcoholic hepatitis: role of Kupffer cells in initiating the process of alcoholic hepatitis; types of leukocytes involved in the pathogenesis of alcoholic hepatitis; chemokines that are responsible for the attraction of leukocytes; adhesion molecules that promote the attachment of leukocytes to the endothelial cells and hepatocytes; mechanisms of leukocyte transmigration to hepatic parenchyma; mechanisms by which leukocytes initiate tissue injury; and interactive effects of alcohol and hepatitis viral proteins on liver injury. This article provides an introduction to the problem and a summary of the 10 scientific presentations delivered at the symposium. (c) 2002 Elsevier Science Inc. All rights reserved. C1 NIAAA, Biomed Res Branch, Div Basic Res, NIH, Bethesda, MD 20892 USA. RP Purohit, V (reprint author), NIAAA, Biomed Res Branch, Div Basic Res, NIH, 6000 Execut Blvd,Suite 402, Bethesda, MD 20892 USA. NR 6 TC 11 Z9 12 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0741-8329 J9 ALCOHOL JI Alcohol PD MAY PY 2002 VL 27 IS 1 BP 3 EP 6 DI 10.1016/S0741-8329(02)00211-2 PG 4 WC Substance Abuse; Pharmacology & Pharmacy; Toxicology SC Substance Abuse; Pharmacology & Pharmacy; Toxicology GA 558YE UT WOS:000175994900013 PM 12062629 ER PT J AU Bautista, AP AF Bautista, AP TI Neutrophilic infiltration in alcoholic hepatitis SO ALCOHOL LA English DT Article; Proceedings Paper CT Annual Meeting of the Research-Society-on-Alcoholism CY JUN 23-28, 2001 CL MONTREAL, CANADA SP Res Soc Alcoholism DE chemokines; cytokines; lymphocytes; macrophages; rats; mice ID MACROPHAGE INFLAMMATORY PROTEIN-2; INTERCELLULAR-ADHESION MOLECULE-1; TUMOR-NECROSIS-FACTOR; KUPFFER CELLS; LIVER-DISEASE; INTERLEUKIN-6 CONCENTRATIONS; KAPPA-B; RAT; INTOXICATION; EXPRESSION AB Leukocyte infiltration in the liver is one of the most important features of alcoholic liver disease. However, in alcoholic hepatitis, the role of polymorphonuclear neutrophils (PMNs) in liver injury still remains to be fully elucidated. Furthermore, the migration of PMNs and their presence in the liver during alcoholic hepatitis have not been fully investigated. Up-regulation of chemokine secretion and adhesion molecule expression on effector cells (i.e., PMNs) and target cells (i.e., hepatocytes) are important factors in neutrophilic infiltration of the liver. The CXC chemokines-that is, interleukin (IL)-8 (in human beings), cytokine -induced neutrophil chemoattractant (CINC) (in rats), and KC (in mice)-are proneutrophilic agents. They are up-regulated during chronic-that is, several years of-alcohol use in human beings and in up to 30 weeks in experimental models of ethanol intoxication in mice and rats. Up-regulation of these chemokines in the circulation and tissues is also associated with enhanced neutrophilic infiltration in the liver. In the rat, the up-regulation of CXC chemokine production is time dependent. For example, after 16 weeks of feeding, up-regulation of CXC chemokine is observed, whereas after 32 weeks, CC chemokines are enhanced. Concomitantly, selective migration of PMNs and mononuclear cells is observed. In another model, in which both CXC and CC chemokines were enhanced after chronic ethanol use for 12 weeks in mice, neutrophilic and mononuclear/lymphocytic infiltrations were also seen. This model correlates closely with alcoholic hepatitis in human beings, characterized by increased IL-8, RANTES (regulated upon activation, normal T cell expressed and secreted), and macrophage inflammatory protein-1 (MIP-1) and profound increases in neutrophils and lymphocytes in the liver. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Louisiana State Univ, Ctr Hlth Sci, Dept Physiol, New Orleans, LA 70112 USA. Louisiana State Univ, Ctr Hlth Sci, NIAAA, Sponsored Alcohol Res Ctr, New Orleans, LA 70112 USA. RP Bautista, AP (reprint author), Louisiana State Univ, Ctr Hlth Sci, Dept Physiol, New Orleans, LA 70112 USA. FU NIAAA NIH HHS [P50 AA09803]; PHS HHS [R01 08846-08] NR 23 TC 75 Z9 77 U1 0 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0741-8329 J9 ALCOHOL JI Alcohol PD MAY PY 2002 VL 27 IS 1 BP 17 EP 21 AR PII S0741-8329(02)00206-9 DI 10.1016/S0741-8329(02)00206-9 PG 5 WC Substance Abuse; Pharmacology & Pharmacy; Toxicology SC Substance Abuse; Pharmacology & Pharmacy; Toxicology GA 558YE UT WOS:000175994900004 PM 12062632 ER PT J AU Gao, B AF Gao, B TI Interaction of alcohol and hepatitis viral proteins: implication in synergistic effect of alcohol drinking and viral hepatitis on liver injury SO ALCOHOL LA English DT Article; Proceedings Paper CT Annual Meeting of the Research-Society-on-Alcoholism CY JUN 23-28, 2001 CL MONTREAL, CANADA SP Res Soc Alcoholism DE alcohol; viral hepatitis; liver injury; hepatitis C virus core protein; hepatitis B virus X protein; NF-kappa B ID C VIRUS-INFECTION; NECROSIS-FACTOR-ALPHA; FACTOR-KAPPA-B; INTERFERON THERAPY; TRANSCRIPTION FACTORS; ACTIVATION; HEPATOCYTES; CONSUMPTION; DISEASE; ETHANOL AB Alcohol drinking and viral hepatitis are both recognized as major causes of liver disease worldwide, and they frequently coexist and synergistically cause liver injury in patients with chronic liver disease. Several mechanisms have been implicated in exacerbation of liver injury in patients with alcohol drinking and viral hepatitis. These include impairment of host defense and liver regeneration by alcohol consumption. The findings obtained from my laboratory have demonstrated that alcohol potentiates cooperatively several signals activated by hepatitis B virus X protein (HBX) or hepatitis C virus core protein, and HBX sensitizes hopatocytes to tumor necrosis factor-alpha (TNF-alpha)- and ethanol-induced apoptosis by a caspase-3-dependent mechanism, which may also contribute to the synergistic effect of alcohol drinking and viral hepatitis on. liver injury. (C) 2002 Elsevier Science Inc. All rights reserved. C1 NIAAA, Sect Liver Biol, Lab Physiol Studies, NIH, Bethesda, MD 20892 USA. RP Gao, B (reprint author), NIAAA, Sect Liver Biol, Lab Physiol Studies, NIH, Pk Bldg,Room 120,12420 Parktown Dr,MSC 8115, Bethesda, MD 20892 USA. NR 36 TC 19 Z9 22 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0741-8329 J9 ALCOHOL JI Alcohol PD MAY PY 2002 VL 27 IS 1 BP 69 EP 72 AR PII S0741-8329(02)00201-X DI 10.1016/S0741-8329(02)00201-X PG 4 WC Substance Abuse; Pharmacology & Pharmacy; Toxicology SC Substance Abuse; Pharmacology & Pharmacy; Toxicology GA 558YE UT WOS:000175994900012 PM 12062640 ER PT J AU Riley, D Berman, B AF Riley, D Berman, B TI Complementary and alternative medicine in outcomes research SO ALTERNATIVE THERAPIES IN HEALTH AND MEDICINE LA English DT Editorial Material ID CARE C1 Univ New Mexico, Sch Med, Albuquerque, NM 87131 USA. Univ Maryland, Sch Med, Complementary Med Program, Baltimore, MD 21201 USA. NIH, Specialized Ctr CAM Res, Bethesda, MD USA. RP Riley, D (reprint author), Univ New Mexico, Sch Med, Albuquerque, NM 87131 USA. OI Riley, David/0000-0001-6903-6624 NR 6 TC 6 Z9 6 U1 0 U2 0 PU INNOVISION COMMUNICATIONS PI ALISO VIEJO PA 101 COLUMBIA, ALISO VIEJO, CA 92656 USA SN 1078-6791 J9 ALTERN THER HEALTH M JI Altern. Ther. Health Med. PD MAY-JUN PY 2002 VL 8 IS 3 BP 36 EP 37 PG 2 WC Integrative & Complementary Medicine SC Integrative & Complementary Medicine GA 550FJ UT WOS:000175492700012 PM 12017498 ER PT J AU Cannon, CP Hand, MH Bahr, R Boden, WE Christenson, R Gibler, WB Eagle, K Lambrew, CT Lee, TH MacLeod, B Ornato, JP Selker, HP Steele, P Zalenski, RJ AF Cannon, CP Hand, MH Bahr, R Boden, WE Christenson, R Gibler, WB Eagle, K Lambrew, CT Lee, TH MacLeod, B Ornato, JP Selker, HP Steele, P Zalenski, RJ CA Natl Heart Attack Alert Program TI Critical pathways for management of patients with acute coronary syndromes: An assessment by the National Heart Attack Alert Program SO AMERICAN HEART JOURNAL LA English DT Article ID ACUTE MYOCARDIAL-INFARCTION; ACUTE CARDIAC ISCHEMIA; RANDOMIZED CONTROLLED TRIAL; CHEST PAIN PATIENT; TIMI-III REGISTRY; UNSTABLE ANGINA; UNITED-STATES; CLINICAL-TRIAL; CATHETERIZATION FACILITIES; CARDIOVASCULAR MEDICINE AB Background The use of critical pathways for a variety of clinical conditions has grown rapidly in recent years, particularly pathways for patients with acute coronary syndromes (ACS). However, no systematic review exists regarding the value of critical pathways in this setting. Methods The National Heart Attack Alert Program established a Working Group to review the utility of critical pathways on quality of care and outcomes for patients with ACS. A literature search of MEDLINE, cardiology textbooks, and cited references in any article identified was conducted regarding the use of critical pathways for patients with ACS. Results Several areas for improving the care of patients with ACS through the application of critical pathways were identified: increasing the use of guideline-recommended medications, targeting use of cardiac procedures and other cardiac testing, and reducing the length of stay in hospitals and intensive care units. Initial studies have shown promising results in improving quality of care and reducing costs. No large studies designed to demonstrate an improvement in mortality or morbidity were identified in this literature review. Conclusions Critical pathways offer the potential to improve the care of patients with ACS while reducing the cost of care. Their use should improve the process and cost-effectiveness of care, but further research in this field is needed to determine whether these changes in the process of care will translate into improved clinical outcomes. C1 NHLBI, Off Prevent Educ & Control, Natl Heart Attack Alert Program, NIH, Bethesda, MD 20892 USA. Brigham & Womens Hosp, Div Cardiovasc, Boston, MA 02115 USA. St Agnes Med Syst, Baltimore, MD USA. Hartford Hosp, Div Cardiol, Hartford, CT 06115 USA. Univ Maryland, Med Ctr, Baltimore, MD 21201 USA. Univ Cincinnati, Coll Med, Dept Emergency Med, Cincinnati, OH USA. Univ Michigan, Med Ctr, Div Cardiol, Ann Arbor, MI 48109 USA. Maine Med Ctr, Div Cardiol, Portland, ME 04102 USA. Partners Community HealthCare Inc, Boston, MA USA. Mercy Hosp, Dept Emergency Med, Pittsburgh, PA 15219 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Emergency Med, Richmond, VA 23298 USA. Tufts Univ New England Med Ctr, Div Clin Care Res, Boston, MA 02111 USA. Vet Hlth Adm, Dept Vet Affairs, Washington, DC USA. Wayne State Univ, Detroit, MI USA. RP Hand, MH (reprint author), NHLBI, Off Prevent Educ & Control, Natl Heart Attack Alert Program, NIH, 31 Ctr Dr,MSC 2480, Bethesda, MD 20892 USA. NR 94 TC 30 Z9 33 U1 0 U2 2 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-8703 J9 AM HEART J JI Am. Heart J. PD MAY PY 2002 VL 143 IS 5 BP 777 EP 789 DI 10.1067/mhj.2002.120260 PG 13 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 560CH UT WOS:000176063800006 PM 12040337 ER PT J AU Holubkov, R Karas, RH Pepine, CJ Rickens, CR Reichek, N Rogers, WJ Sharaf, BL Sopko, G Merz, CNB Kelsey, SF McGorray, SP Reis, SE AF Holubkov, R Karas, RH Pepine, CJ Rickens, CR Reichek, N Rogers, WJ Sharaf, BL Sopko, G Merz, CNB Kelsey, SF McGorray, SP Reis, SE TI Large brachial artery diameter is associated with angiographic coronary artery disease in women SO AMERICAN HEART JOURNAL LA English DT Article ID COMMUNITIES ARIC COHORT; ENDOTHELIAL DYSFUNCTION; ATHEROSCLEROSIS RISK; ENLARGEMENT; ADULTS AB Background Noninvasive methods are needed for the identification of women at highest risk for coronary artery disease (CAD) who might benefit most from aggressive preventive therapy. Identification of brachial artery atherosclerosis, which correlates with coronary artery atherosclerosis, may be useful to estimate or stratify CAD risk. Because atherosclerosis disrupts the arterial architecture that regulates vessel size, we hypothesized that noninvasively measured large brachial artery diameter is a manifestation of atherosclerosis that is associated with angiographic CAD in women with chest pain. Methods We examined 376 women (mean age, 57.1 years) with chest pain in the National Heart, Lung, and Blood Institute's Women's Ischemia Syndrome Evaluation study who underwent B-mode ultrasound scan measurement of brachial artery diameter at rest and during hyperemic stress (to quantify flow-mediated dilation), quantitative coronary angiography, and risk factor assessment. Results Large resting brachial artery diameter was associated with significant angiographic CAD (3.90 +/- 0.79 mm vs 3.52 +/- 0.59 mm in women with CAD vs no CAD; P < .001). Impaired flow-mediated dilation, which correlated with resting diameter (r = -0. 17; P = .001), was weakly associated with significant CAD (2.74% +/- 7.11% vs 4.48% +/- 9.52% in CAD vs no CAD; P = .046). After adjustment for age, body size, and CAD risk factors, women with large resting brachial artery diameters (>4.1 mm) had 3.6-fold increased odds (95% confidence interval, 1.8 to 7.1; P < .001) of significant angiographic CAD compared with those with small brachial arteries (less than or equal to3.6 mm). Conclusion Large resting brachial artery diameter is an independent predictor of significant CAD in women with chest pain. Therefore, a simple ultrasonographic technique may be useful in the identification of women with chest pain who are at increased risk for CAD. C1 Univ Pittsburgh, Med Ctr, Dept Epidemiol, Pittsburgh, PA 15213 USA. Univ Pittsburgh, Cardiovasc Inst, Pittsburgh, PA 15213 USA. Tufts Univ, New England Med Ctr, Mol Cardiol Res Ctr, Boston, MA 02111 USA. Univ Florida, Div Cardiol, Gainesville, FL USA. Allegheny Gen Hlth Syst, Div Cardiol, Birmingham, AL USA. Univ Alabama, Div Cardiol, Birmingham, AL USA. Rhode Isl Hosp, Div Cardiol, Bethesda, MD USA. NHLBI, Div Heart & Vasc Dis, Los Angeles, CA USA. Cedars Sinai Med Ctr, Div Cardiol, Los Angeles, CA 90048 USA. RP Reis, SE (reprint author), Univ Pittsburgh, Med Ctr, Dept Epidemiol, 200 Lothrop St, Pittsburgh, PA 15213 USA. RI Reis, Steven/J-3957-2014 FU NCRR NIH HHS [M01-RR00425]; NHLBI NIH HHS [N01-HV-68161, N01-HV-68162, N01-HV-68163, N01-HV-68164] NR 20 TC 56 Z9 58 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-8703 J9 AM HEART J JI Am. Heart J. PD MAY PY 2002 VL 143 IS 5 BP 802 EP 807 DI 10.1067/mhj.2002.121735 PG 6 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 560CH UT WOS:000176063800009 PM 12040340 ER PT J AU Geleijnse, JM Launer, LJ van der Kuip, DAM Hofman, A Witteman, JCM AF Geleijnse, JM Launer, LJ van der Kuip, DAM Hofman, A Witteman, JCM TI Inverse association of tea and flavonoid intakes with incident myocardial infarction: the Rotterdam Study SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Article DE tea; flavonoids; myocardial infarction; ischemic heart disease; population-based study; the Rotterdam Study; Netherlands ID CORONARY-HEART-DISEASE; ANTIOXIDANT ACTIVITY; ARTERY DISEASE; NITRIC-OXIDE; QUERCETIN; HEALTH; RISK; LDL; CONSUMPTION; CHOLESTEROL AB Background: Dietary flavonoids may protect against cardiovascular disease, but evidence is still conflicting. Tea is the major source of flavonoids in Western populations. Objective: The association of tea and flavonoid intake with incident myocardial infarction was examined in the general Dutch population. Design: A longitudinal analysis was performed with the use of data from the Rotterdam Study-a population-based study of men and women aged greater than or equal to55 y. Diet was assessed at baseline (1990-1993) with a validated semiquantitative food-frequency questionnaire. The analysis included 4807 subjects with no history of myocardial infarction, who were followed until 31 December 1997. Data were analyzed in a Cox regression model, with adjustment for age, sex, body mass index, smoking status, pack-years of cigarette smoking, education level, and daily intakes of alcohol, coffee, polyunsaturated fat, saturated fat, fiber, vitamin E, and total energy. Results: During 5.6 y of follow-up, a total of 146 first myocardial infarctions occurred, 30 of which were fatal. The relative risk (RR) of incident myocardial infarction was lower in tea drinkers with a daily intake >375 mL (RR: 0.57; 95% CI: 0.33, 0.98) than in nontea drinkers. The inverse association with tea drinking was stronger for fatal events (0.30; 0.09, 0.94) than for nonfatal events (0.68; 0.37, 1.26). The intake of dietary flavonoids (quercetin + kaempferol + myricetin) was significantly inversely associated only with fatal myocardial infarction (0.35, 0.13, 0.98) in upper compared with lower tertiles of intake. Conclusions: An increased intake of tea and flavonoids may contribute to the primary prevention of ischemic heart disease. C1 Erasmus Med Ctr, Dept Epidemiol & Biostat, NL-3000 DR Rotterdam, Netherlands. Wageningen Univ, Div Human Nutr & Epidemiol, Wageningen, Netherlands. Natl Inst Aging, Bethesda, MD USA. RP Witteman, JCM (reprint author), Erasmus Med Ctr, Dept Epidemiol & Biostat, POB 1738, NL-3000 DR Rotterdam, Netherlands. RI Geleijnse, Johanna/B-5749-2012 NR 37 TC 266 Z9 277 U1 1 U2 45 PU AMER SOC CLINICAL NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, SUBSCRIPTIONS, RM L-3300, BETHESDA, MD 20814-3998 USA SN 0002-9165 J9 AM J CLIN NUTR JI Am. J. Clin. Nutr. PD MAY PY 2002 VL 75 IS 5 BP 880 EP 886 PG 7 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 545CA UT WOS:000175197600013 PM 11976162 ER PT J AU Flood, A Velie, EM Chaterjee, N Subar, AF Thompson, FE Lacey, JV Schairer, C Troisi, R Schatzkin, A AF Flood, A Velie, EM Chaterjee, N Subar, AF Thompson, FE Lacey, JV Schairer, C Troisi, R Schatzkin, A TI Fruit and vegetable intakes and the risk of colorectal cancer in the Breast Cancer Detection Demonstration Project follow-up cohort SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Article DE fruit; vegetables; colorectal cancer; prospective study; women; energy adjustment; Breast Cancer Detection Demonstration Project ID UNITED-STATES; COLON-CANCER; DIET; CONSUMPTION AB Background: Recent findings have cast doubt on the hypothesis that high intakes of fruit and vegetables are associated with a reduced risk of colorectal cancer. Objective: In a large prospective cohort of women, we examined the association between fruit and vegetable intakes and colorectal cancer. Design: Between 1987 and 1989, 45490 women with no history of colorectal cancer satisfactorily completed a 62-item Block-National Cancer Institute food-frequency questionnaire. During 386 142 person-years of follow-up, 314 women reported incident colorectal cancer, searches of the National Death Index identified an additional 106 colorectal cancers, and a match with state registries identified another 65 colorectal cancers for a total of 485 cases. We used Cox proportional hazards regression analysis to estimate the relative risks (RRs) and 95% CIs in both energy-adjusted and fully adjusted models. Results: In models using the multivariate nutrient-density model of energy adjustment, RRs for increasing quintile of fruit consumption indicated no significant association with colorectal cancer [RR (95% CI)]: 1.00 (reference), 0.94 (0.70, 1.26), 0.85 (0.63, 1.15), 1.07 (0.81, 1.42), and 1.09 (0.82, 1.44). For vegetable consumption, there was also no significant association in the multivariate nutrient-density model with increasing quintiles of consumption: 1.00 (reference), 0.77 (0.58, 1.02), 0.83 (0.63, 1.10), 0.90 (0.69, 1.19), and 0.92 (0.70, 1.22). Additionally, 3 alternative models of energy adjustment showed no significant association between increases in vegetable intake and the risk of colorectal cancer. Conclusion: Although the Limitations of our study design and data merit consideration, this investigation provides little evidence of an association between fruit and vegetable intakes and colorectal cancer. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Michigan State Univ, Dept Epidemiol, E Lansing, MI 48824 USA. RP Flood, A (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,MSC 7232, Bethesda, MD 20892 USA. NR 26 TC 89 Z9 90 U1 3 U2 9 PU AMER SOC CLINICAL NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, SUBSCRIPTIONS, RM L-3300, BETHESDA, MD 20814-3998 USA SN 0002-9165 J9 AM J CLIN NUTR JI Am. J. Clin. Nutr. PD MAY PY 2002 VL 75 IS 5 BP 936 EP 943 PG 8 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 545CA UT WOS:000175197600021 PM 11976170 ER PT J AU Brenner, RA Scheidt, PC Rossi, MW Cheng, TL Overpeck, MD Boenning, DA Wright, JL Kavee, JD Boyle, KE AF Brenner, RA Scheidt, PC Rossi, MW Cheng, TL Overpeck, MD Boenning, DA Wright, JL Kavee, JD Boyle, KE TI Injury surveillance in the ED: Design, implementation, and analysis SO AMERICAN JOURNAL OF EMERGENCY MEDICINE LA English DT Article DE population surveillance; emergency department; data collection ID EMERGENCY DEPARTMENTS; PREVENTION; TELEPHONE; ACCIDENT; EPIDEMIOLOGY; ADOLESCENTS; INFORMATION; CHALLENGES; CHILDREN; PROGRAM AB Comprehensive, population-based surveillance for nonfatal injuries requires uniform methods for data collection from multiple hospitals. To show issues related to design and implementation of multihospital, emergency department (ED), injury surveillance, a city-wide system in the United States is discussed. From October 1, 1995 to September 30, 1996 all injury-related ED visits among District of Columbia residents <3 years of age were ascertained at the 10 hospitals where city children routinely sought care. Information was abstracted from 2,938 injury-related, ED visits (132.7 visits/1,000 person-years). Based on this experience, suggestions to facilitate design of multihospital, injury surveillance in other locations are offered. Importantly, injury-related visits were reliably ascertained from ED logs, and for most variables, a systematic sample of injury-related visits was representative of the total injured population. However, there is a need for more complete documentation of circumstances surrounding injuries and for standardization of data elements on ED logs and treatment records. (Am J Emerg Med 2002;20:181-187. This is a US government work. There are no restrictions on its use.) C1 NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. Maternal & Child Hlth Bur, Rockville, MD USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. RP Brenner, RA (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, 6100 Execut Bldg,Room 7B03, Bethesda, MD 20892 USA. FU NICHD NIH HHS [U18-HD30458, U18-HD31919, U18-HD31206, U18-HD30455, U18-HD30454, U18-HD30450, U18-HD30477] NR 56 TC 8 Z9 8 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 0735-6757 J9 AM J EMERG MED JI Am. J. Emerg. Med. PD MAY PY 2002 VL 20 IS 3 BP 181 EP 187 DI 10.1053/ajem.2002.32639 PG 7 WC Emergency Medicine SC Emergency Medicine GA 549WK UT WOS:000175469600009 PM 11992337 ER PT J AU Stolzenberg-Solomon, RZ Pietinen, P Taylor, PR Virtamo, J Albanes, D AF Stolzenberg-Solomon, RZ Pietinen, P Taylor, PR Virtamo, J Albanes, D TI Prospective study of diet and pancreatic cancer in male smokers SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE carbohydrates; diet; dietary fats; energy intake; pancreatic neoplasms; prospective studies; smoking ID NUTRITIONAL FACTORS; ORGANOCHLORINE COMPOUNDS; FRANCOPHONE COMMUNITY; NUTRIENT INTAKE; RISK-FACTORS; CARCINOGENESIS; FAT; RAT; CARCINOMA; MONTREAL AB There have been few prospective studies relating diet to pancreatic cancer, with most having fewer than 100 cases and only one examining dietary nutrients. The authors prospectively examined dietary factors hypothesized to be associated with exocrine pancreatic cancer in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort in Finland. Of the 27,111 male smokers aged 50-69 years with complete dietary information, as ascertained from a self-administered dietary history questionnaire given at baseline (1985-1988), 163 developed pancreatic cancer from 1985 through November 1997. Cox proportional hazards models were used to estimate smoking- and age-adjusted hazard ratios and 95% confidence intervals. Energy-adjusted butter consumption and saturated fat intake were positively associated with pancreatic cancer (highest quintile vs. lowest: hazard ratio (HR) = 1.40, 95% confidence interval (CI): 0.87, 2.25 (p trend = 0.04), and HR = 1.60, 95% CI: 0.96, 2.64 (p trend = 0.02), respectively). Energy intake and energy-adjusted carbohydrate intake were inversely associated with the disease (highest quintile vs. lowest: HR = 0.62, 95% CI: 0.36, 1.07 (p trend = 0.05), and HR = 0.62, 95% CI: 0.37, 1.03 (p trend = 0.02), respectively). These results support the hypothesis that a high intake of saturated fat may increase the risk of pancreatic cancer in smokers, while greater intakes of energy and carbohydrate may reduce the risk. C1 NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Natl Publ Hlth Inst, Helsinki, Finland. NCI, Canc Prevent Studies Branch, Ctr Canc Res, Bethesda, MD 20892 USA. RP Stolzenberg-Solomon, RZ (reprint author), NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, 6006 Execut Blvd,MSC 7232, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 FU NCI NIH HHS [N01CN45035, N01CN45165] NR 58 TC 147 Z9 149 U1 2 U2 7 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD MAY 1 PY 2002 VL 155 IS 9 BP 783 EP 792 DI 10.1093/aje/155.9.783 PG 10 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 548PJ UT WOS:000175397300001 PM 11978580 ER PT J AU Stolzenberg-Solomon, RZ Pietinen, P Taylor, PR Virtamo, J Albanes, D AF Stolzenberg-Solomon, RZ Pietinen, P Taylor, PR Virtamo, J Albanes, D TI Stolzenberg-Solomon et al. respond to "What do we know about pancreas cancer?" by Potter SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Editorial Material ID MALE SMOKERS; RISK FACTOR; VALIDITY; DIETARY; CARCINOGENESIS; NUTRITION; DISEASE; COHORT; FAT C1 NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Natl Publ Hlth Inst, Helsinki, Finland. NCI, Canc Prevent Studies Branch, Ctr Canc Res, Bethesda, MD 20892 USA. RP Stolzenberg-Solomon, RZ (reprint author), NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, 6120 Execut Bldg,MSC 7232, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 NR 17 TC 1 Z9 1 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD MAY 1 PY 2002 VL 155 IS 9 BP 796 EP 797 DI 10.1093/aje/155.9.796 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 548PJ UT WOS:000175397300003 ER PT J AU Groves, FD Page, WF Gridley, G Lisimaque, L Stewart, PA Tarone, RE Gail, MH Boice, JD Beebe, GW AF Groves, FD Page, WF Gridley, G Lisimaque, L Stewart, PA Tarone, RE Gail, MH Boice, JD Beebe, GW TI Cancer in Korean War navy technicians: Mortality survey after 40 years SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE leukemia, nonlymphocytic, acute; microwaves; mortality; neoplasms; veterans ID CELLULAR-TELEPHONE USE; BRAIN-TUMORS; EXPOSURE; RADIATION; RISK; ASCERTAINMENT; OCCUPATIONS; MELANOMA; VETERANS; PHONES AB This study reports on over 40 years of mortality follow-up of 40,581 Navy veterans of the Korean War with potential exposure to high-intensity radar. The cohort death rates were compared with mortality rates for White US men using standardized mortality ratios, and the death rates for men in occupations considered a priori to have high radar exposure were compared with the rates for men in low-exposure occupations using Poisson regression. Deaths from all diseases and all cancers were significantly below expectation overall and for the 20,021 sailors with high radar exposure potential. There was no evidence of increased brain cancer in the entire cohort (standardized mortality ratio (SMR) = 0.9, 95% confidence interval (CI): 0.7, 1.1) or in high-exposure occupations (SMR = 0.7, 95% CI: 0.5, 1.0). Testicular cancer deaths also occurred less frequently than expected in the entire cohort and high-exposure occupations. Death rates for several smoking-related diseases were significantly lower in the high-exposure occupations. Nonlymphocytic leukemia was significantly elevated among men in high-exposure occupations but in only one of the three high-exposure occupations, namely, electronics technicians in aviation squadrons (SMR = 2.2, 95% CI: 1.3, 3.7). Radar exposure had little effect on mortality in this cohort of US Navy veterans. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Natl Acad Sci, Natl Res Council, Med Follow Up Agcy, Washington, DC 20418 USA. Vanderbilt Univ, Vanderbilt Ingram Canc Ctr, Nashville, TN USA. Int Epidemiol Inst, Rockville, MD USA. RP Groves, FD (reprint author), Med Univ S Carolina, Dept Biometry & Epidemiol, 135 Cannon St,Room 302-H, Charleston, SC 29425 USA. NR 24 TC 40 Z9 43 U1 0 U2 2 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD MAY 1 PY 2002 VL 155 IS 9 BP 810 EP 818 DI 10.1093/aje/155.9.810 PG 9 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 548PJ UT WOS:000175397300005 PM 11978584 ER PT J AU Falanga, A Toma, S Marchetti, M Palumbo, R Raffo, P Consonni, R Marziali, S Dastoli, G Barbui, T AF Falanga, A Toma, S Marchetti, M Palumbo, R Raffo, P Consonni, R Marziali, S Dastoli, G Barbui, T TI Effect of all-trans-retinoic acid on the hypercoagulable state of patients with breast cancer SO AMERICAN JOURNAL OF HEMATOLOGY LA English DT Article DE hypercoagulation; breast cancer patients; ATRA ID ACUTE PROMYELOCYTIC LEUKEMIA; HUMAN ENDOTHELIAL-CELLS; ALPHA-INTERFERON 2A; LOW-DOSE WARFARIN; PROCOAGULANT ACTIVITY; TISSUE-FACTOR; VENOUS THROMBOEMBOLISM; PLASMINOGEN-ACTIVATOR; POSTMENOPAUSAL WOMEN; THROMBOPHILIC STATE AB To evaluate whether all-trans-retinoic acid (ATRA) is able to modulate the hemostatic system in patients with solid tumors, we studied patients with locally advanced breast cancer who were enrolled in a Phase lb study of ATRA Tamoxifen (Tam). In this study, two groups of 15 patients/each were treated for 21 days before operation with ATRA at three doses (15, 45, or 75 mg/m(2)/day on alternate days) given alone (group 1) or in combination with Tam (group 2). One additional group received Tam alone. Plasma samples were evaluated for hypercoagulation markers (FVIIa, F1+2, TAT, D-dimer), fibrinolysis proteins (t-PA, PAI-1), and coagulation inhibitors (protein C, AT). At baseline, cancer patients had FVIIa, F1+2, TAT, and PAI-1 significantly greater than control subjects. During treatment, in the patients given ATRA alone, hypercoagulation markers appeared unmodified. Instead, subjects given Tam alone had a significant elevation of FVIIa, F1+2, and TAT versus baseline. However, in the ATRA + Tam groups, hypercoagulation markers were decreased compared with Tam alone. These results suggest that in selected conditions, pre-operative ATRA may modulate the hypercoagulable state of breast cancer patients. (C) 2002 Wiley-Liss, Inc. C1 Osped Ruiniti, Dept Hematol, I-14128 Bergamo, Italy. Univ Genoa, Natl Canc Inst, Dept Oncol Biol & Genet, Genoa, Italy. Roche SpA, Monza, Italy. RP Falanga, A (reprint author), Osped Ruiniti, Dept Hematol, Largo Barozzi, I-14128 Bergamo, Italy. OI Falanga, Anna/0000-0001-6906-9020 NR 46 TC 8 Z9 8 U1 2 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0361-8609 J9 AM J HEMATOL JI Am. J. Hematol. PD MAY PY 2002 VL 70 IS 1 BP 9 EP 15 DI 10.1002/ajh.10073 PG 7 WC Hematology SC Hematology GA 548TP UT WOS:000175406200002 PM 11994976 ER PT J AU Rokman, A Ikonen, T Seppala, EH Nupponen, N Autio, V Mononen, N Bailey-Wilson, J Trent, J Carpten, J Matikainen, MP Koivisto, PA Tammela, TLJ Kallioniemi, OP Schleutker, J AF Rokman, A Ikonen, T Seppala, EH Nupponen, N Autio, V Mononen, N Bailey-Wilson, J Trent, J Carpten, J Matikainen, MP Koivisto, PA Tammela, TLJ Kallioniemi, OP Schleutker, J TI Germline alterations of the RNASEL gene, a candidate HPC1 gene at 1q25, in patients and families with prostate cancer SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID 2-5A-DEPENDENT RNASE; INTERFERON AB The RNASEL gene (2',5'-oligoisoadenylate-synthetase dependent) encodes a ribonuclease that mediates the antiviral and apoptotic activities of interferons. The RNASEL gene maps to the hereditary-prostate-cancer (HPC)-predisposition locus at 1q24-q25 (HPC1) and was recently shown to harbor truncating mutations in two families with linkage to HPC1. Here, we screened for RNASEL germline mutations in 66 Finnish patients with HPC, and we determined the frequency of the changes in the index patients from 116 families with HPC, in 492 patients with unselected prostate cancer (PRCA), in 223 patients with benign prostatic hyperplasia (BPH), and in 566 controls. A truncating mutation, E265X, was found in 5 (4.3%) of the 116 patients from families with HPC. This was significantly higher (odds ratio [OR] = 4.56 P = .04) than the frequency of E265X in controls (1.8%). The highest mutation frequency (9.5%) was found in patients from families with four or more affected members. Possible segregation was detected only in a single family. However, the median age at disease onset for E265X carriers was 11 years less than that for noncarriers in the same families. In addition, of the four missense variants found, R462Q showed an association with HPC (OR = 1.96; P = .07). None of the variants showed any differences between controls and either patients with BPH or patients with PRCA. We conclude that, although RNASEL mutations do not explain disease segregation in Finnish families with HPC, the variants are enriched in families with HPC that include more than two affected members and may also be associated with the age at disease onset. This suggests a possible modifying role in cancer predisposition. The impact that the RNASEL sequence variants have on PRCA burden at the population level seems small but deserves further study. C1 Univ Tampere, Inst Med Technol, Canc Genet Lab, FIN-33014 Tampere, Finland. Univ Tampere, Tampere Sch Publ Hlth, FIN-33101 Tampere, Finland. Univ Tampere, Sch Med, Dept Urol, FIN-33101 Tampere, Finland. Tampere Univ Hosp, Tampere, Finland. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. NIH, Ctr Inherited Dis Res, Baltimore, MD USA. RP Rokman, A (reprint author), Univ Tampere, Inst Med Technol, Canc Genet Lab, FIN-33014 Tampere, Finland. RI Kallioniemi, Olli/H-5111-2011; Kallioniemi, Olli/H-4738-2012; OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332; Bailey-Wilson, Joan/0000-0002-9153-2920 FU NHGRI NIH HHS [N01-HG-55389] NR 21 TC 136 Z9 140 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD MAY PY 2002 VL 70 IS 5 BP 1299 EP 1304 DI 10.1086/340450 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 541XT UT WOS:000175012400019 PM 11941539 ER PT J AU Richard, G Rouan, F Willoughby, CE Brown, N Chung, P Ryynanen, M Jabs, EW Bale, SJ DiGiovanna, JJ Uitto, J Russell, L AF Richard, G Rouan, F Willoughby, CE Brown, N Chung, P Ryynanen, M Jabs, EW Bale, SJ DiGiovanna, JJ Uitto, J Russell, L TI Missense mutations in GJB2 encoding connexin-26 cause the ectodermal dysplasia keratitis-ichthyosis-deafness syndrome SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID GAP JUNCTION PROTEINS; KID-SYNDROME; ERYTHROKERATODERMIA VARIABILIS; CHANNELS; CELLS; KERATODERMA; EXPRESSION; FAMILY; GENE AB Keratitis-ichthyosis-deafness syndrome (KID) is a rare ectodermal dysplasia characterized by vascularizing keratitis, profound sensorineural hearing loss (SNHL), and progressive erythrokeratoderma, a clinical triad that indicates a failure in development and differentiation of multiple stratifying epithelia. Here, we provide compelling evidence that KID is caused by heterozygous missense mutations in the connexin-26 gene, GJB2. In each of 10 patients with KID, we identified a point mutation leading to substitution of conserved residues in the cytoplasmic amino terminus or first extracellular domain of Cx26. One of these mutations was detected in six unrelated sporadic case subjects and also segregated in one family with vertical transmission of KID. These results indicate the presence of a common, recurrent mutation and establish its autosomal dominant nature. Cx26 and the closely related Cx30 showed differential expression in epidermal, adnexal, and corneal epithelia but were not significantly altered in lesional skin. However, mutant Cx26 was incapable of inducing intercellular coupling in vitro, which indicates its functional impairment. Our data reveal striking genotype-phenotype correlations and demonstrate that dominant GJB2 mutations can disturb the gap junction system of one or several ectodermal epithelia, thereby producing multiple phenotypes: nonsyndromic SNHL, syndromic SNHL with palmoplantar keratoderma, and KID. Decreased host defense and increased carcinogenic potential in KID illustrate that gap junction communication plays not only a crucial role in epithelial homeostasis and differentiation but also in immune response and epidermal carcinogenesis. C1 Thomas Jefferson Univ, Dept Dermatol & Cutaneous Biol, BLSB, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Jefferson Inst Mol Med, Philadelphia, PA 19107 USA. Univ Liverpool, Dept Med, Expt Ophthalmol Unit, Liverpool L69 3BX, Merseyside, England. Oulu Univ Hosp, Dept Obstet & Gynecol, Oulu, Finland. Johns Hopkins Univ Hosp, Dept Pediat Med & Plast Surg, Inst Med Genet, Baltimore, MD 21287 USA. NIAMSD, Genet Studies Sect, NIH, Bethesda, MD 20892 USA. GeneDx, Rockville, MD USA. Brown Univ, Rhode Isl Hosp, Dept Dermatol, Providence, RI 02903 USA. Montreal Childrens Hosp, Div Med Genet, Montreal, PQ H3H 1P3, Canada. RP Richard, G (reprint author), Thomas Jefferson Univ, Dept Dermatol & Cutaneous Biol, BLSB, Suite 450,233 S 10th St, Philadelphia, PA 19107 USA. OI Jabs, Ethylin/0000-0001-8983-5466 FU NIAMS NIH HHS [P01 AR038923, K08 AR002141, P01-AR38923, K08-AR02141] NR 30 TC 220 Z9 240 U1 0 U2 5 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD MAY PY 2002 VL 70 IS 5 BP 1341 EP 1348 DI 10.1086/339986 PG 8 WC Genetics & Heredity SC Genetics & Heredity GA 541XT UT WOS:000175012400024 PM 11912510 ER PT J AU Arikawa-Hirasawa, E Le, AH Nishino, I Nonaka, I Ho, NC Francomano, CA Govindraj, P Hassell, JR Devaney, JM Spranger, J Stevenson, RE Iannaccone, S Dalakas, MC Yamada, Y AF Arikawa-Hirasawa, E Le, AH Nishino, I Nonaka, I Ho, NC Francomano, CA Govindraj, P Hassell, JR Devaney, JM Spranger, J Stevenson, RE Iannaccone, S Dalakas, MC Yamada, Y TI Structural and functional mutations of the perlecan gene cause Schwartz-Jampel syndrome, with myotonic myopathy and chondrodysplasia SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID HEPARAN-SULFATE PROTEOGLYCAN; ACETYLCHOLINESTERASE DEFICIENCY; CHONDRODYSTROPHIC MYOTONIA; BASEMENT-MEMBRANES; MYASTHENIC SYNDROME; CELL-ADHESION; DOMAIN-V; CARTILAGE; FIBULIN-2; RECEPTOR AB Perlecan, a large heparan sulfate proteoglycan, is a component of the basement membrane and other extracellular matrices and has been implicated in multiple biological functions. Mutations in the perlecan gene (HSPG2) cause two classes of skeletal disorders: the relatively mild Schwartz-Jampel syndrome (SJS) and severe neonatal lethal dyssegmental dysplasia, Silverman-Handmaker type (DDSH). SJS is an autosomal recessive skeletal dysplasia characterized by varying degrees of myotonia and chondrodysplasia, and patients with SJS survive. The molecular mechanism underlying the chondrodystrophic myotonia phenotype of SJS is unknown. In the present report, we identify five different mutations that resulted in various forms of perlecan in three unrelated patients with SJS. Heterozygous mutations in two patients with SJS either produced truncated perlecan that lacked domain V or significantly reduced levels of wild-type perlecan. The third patient had a homozygous 7-kb deletion that resulted in reduced amounts of nearly full-length perlecan. Unlike DDSH, the SJS mutations result in different forms of perlecan in reduced levels that are secreted to the extracellular matrix and are likely partially functional. These findings suggest that perlecan has an important role in neuromuscular function and cartilage formation, and they define the molecular basis involved in the difference in the phenotypic severity between DDSH and SJS. C1 Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, Bethesda, MD USA. NINCDS, Bethesda, MD 20892 USA. Natl Inst Neurosci, Dept Ultrastruct Res, Tokyo, Japan. NIA, Genet Lab, Baltimore, MD 21224 USA. Johns Hopkins Bayview Med Ctr, Baltimore, MD USA. Shriners Hosp Children, Tampa, FL USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. Greenwood Genet Ctr, Greenwood, SC 29646 USA. Texas Scottish Rite Hosp Children, Dallas, TX 75219 USA. RP Yamada, Y (reprint author), NIDCR, NIH, Bldg 30,Room 405,30 Convent Dr, Bethesda, MD 20892 USA. OI Nishino, Ichizo/0000-0001-9452-112X NR 32 TC 83 Z9 90 U1 0 U2 3 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD MAY PY 2002 VL 70 IS 5 BP 1368 EP 1375 DI 10.1086/340390 PG 8 WC Genetics & Heredity SC Genetics & Heredity GA 541XT UT WOS:000175012400028 PM 11941538 ER PT J AU Coresh, J Astor, BC McQuillan, G Kusek, J Greene, T Van Lente, F Levey, AS AF Coresh, J Astor, BC McQuillan, G Kusek, J Greene, T Van Lente, F Levey, AS TI Calibration and random variation of the serum creatinine assay as critical elements of using equations to estimate glomerular filtration rate SO AMERICAN JOURNAL OF KIDNEY DISEASES LA English DT Article DE glomerular filtration rate (GFR); prediction equations; serum creatinine ID CHRONIC RENAL-DISEASE; CLEARANCE; PREDICTION; AMERICAN AB Equations using serum creatinine level, age, sex, and other patient characteristics often are used to estimate glomerular filtration rate (GFR) in both clinical practice and research studies. However, the critical dependence of these equations on serum creatinine assay calibration often is overlooked, and the reproducibility of estimated GFR is rarely discussed. We address these issues in frozen samples from 212 Modification of Diet in Renal Disease (MDRD) study participants and 342 Third National Health and Nutrition Examination Survey (NHANES 111) participants assayed for serum creatinine level a second time during November 2000. Variation in serum creatinine level was assessed in 1,919 NHANES III participants who had serum creatinine measured on two visits a median of 17 days apart. Linear regression was used to compare estimates. Calibration of serum creatinine varied substantially across laboratories and time. Data indicate that serum creatinine assays on the same samples were 0.23 mg/dL higher in the NHANES III than MDRD study. Data from the College of American Pathologists suggest that a difference of this magnitude across laboratories is not unusual. Conversely, serum creatinine assays an average of 2 weeks apart have better precision (SD of percentage of difference in estimated GFR, 15%; 90% of estimates within 21%). Errors in calibration make little difference in estimating severely decreased GFR (<30 mL/min/1.73 m(2)), but result in progressively larger differences at higher GFRs. Both clinical and research use of serum creatinine or equations to estimate GFR require knowledge of the calibration of the serum creatinine assay. (C) 2002 by the National Kidney Foundation, Inc. C1 Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Epidemiol, Baltimore, MD 21218 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Biostat, Baltimore, MD 21218 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21218 USA. Johns Hopkins Med Inst, Welch Ctr Prevent Epidemiol & Clin Res, Baltimore, MD 21205 USA. Ctr Dis Control & Prevent, Div Hlth Examinat Stat, Natl Ctr Hlth Stat, Hyattsville, MD 20782 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. Cleveland Clin, Res Fdn, Cleveland, PA USA. Tufts Univ, New England Med Ctr Hosp, Sch Med, Dept Med,Div Nephrol, Boston, MA USA. RP Coresh, J (reprint author), 2024 E Monument, Baltimore, MD 21205 USA. FU NCRR NIH HHS [RR00722]; NHLBI NIH HHS [T32HL7024]; NIDDK NIH HHS [DK48362, R01 DK53869, UO1 DK35073] NR 14 TC 518 Z9 527 U1 1 U2 4 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0272-6386 J9 AM J KIDNEY DIS JI Am. J. Kidney Dis. PD MAY PY 2002 VL 39 IS 5 BP 920 EP 929 DI 10.1053/ajkd.2002.32765 PG 10 WC Urology & Nephrology SC Urology & Nephrology GA 548JQ UT WOS:000175385200002 PM 11979335 ER PT J AU Tveit, DP Hypolite, IO Hshieh, P Cruess, D Agodoa, LY Welch, PG Abbott, KC AF Tveit, DP Hypolite, IO Hshieh, P Cruess, D Agodoa, LY Welch, PG Abbott, KC TI Chronic dialysis patients have high risk for pulmonary embolism SO AMERICAN JOURNAL OF KIDNEY DISEASES LA English DT Article DE pulmonary embolism; dialysis; peritoneal dialysis (PD); hemodialysis (HD); complications; US Renal Data System (USRDS); National Center for Health Statistics (NCHS) ID STAGE RENAL-DISEASE; ACTIVATED-PROTEIN-C; MYOCARDIAL-INFARCTION; UREMIC PATIENTS; PLASMA-LEVELS; FAILURE; ABNORMALITIES; COMPLICATIONS; HEMODIALYSIS; MORTALITY AB Pulmonary embolism has been considered uncommon in chronic dialysis patients, but has not been adequately studied in a large population. In the US Renal Data System (USRDS), 76,718 patients presenting with end-stage renal disease (ESRD) between January 1, 1996, and December 31,1996, were analyzed in an historical cohort study. The outcome was hospitalizations with a primary discharge diagnosis of pulmonary embolism (international Classification of Diseases, Ninth Revision code 415.1x) occurring within 1 year of the first ESRD treatment and excluding those occurring after renal transplantation. For dialysis patients, hospitalization rates for pulmonary embolism were obtained from the hospitalization section of the 1999 USRDS. For the general population, hospitalization rates for pulmonary embolism were obtained from the National Hospital Discharge Survey for 1996. Comorbidities from the Medical Evidence Form (Centers for Medicare and Medicaid Services, previously known as the Health Care Financing Administration; form 2728) were used to generate approximated stratified models of adjusted incidence ratios for pulmonary embolism (comorbidities could not be stratified for the general population). In 1996, the overall incidence rate of pulmonary embolism was 149.90/100,000 dialysis patients compared with 24.62/100,000 persons in the US population, with an age-adjusted incidence ratio of 2.34 in dialysis patients. Younger dialysis patients had the greatest relative risk for pulmonary embolism. The age-adjusted incidence ratio of pulmonary embolism after excluding dialysis patients with known risk factors for pulmonary embolism was 2.11. Ninety-five percent confidence intervals for all age categories in both models were statistically significant. Chronic dialysis patients have high risk for pulmonary embolism, independent of comorbidity. This is a US government work. There are no restrictions on its use. C1 Walter Reed Army Med Ctr, Serv Nephrol, Washington, DC 20307 USA. Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA. NIDDKD, NIH, Off Minor Hlth Res Coordinat, Bethesda, MD USA. RP Abbott, KC (reprint author), Walter Reed Army Med Ctr, Serv Nephrol, Washington, DC 20307 USA. OI Abbott, Kevin/0000-0003-2111-7112 NR 35 TC 67 Z9 69 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 0272-6386 J9 AM J KIDNEY DIS JI Am. J. Kidney Dis. PD MAY PY 2002 VL 39 IS 5 BP 1011 EP 1017 DI 10.1053/ajkd.2002.32774 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA 548JQ UT WOS:000175385200011 PM 11979344 ER PT J AU Stoner, GL Alappan, R Jobes, DV Ryschkewitsch, CF Landry, ML AF Stoner, GL Alappan, R Jobes, DV Ryschkewitsch, CF Landry, ML TI BK virus regulatory region rearrangements in brain and cerebrospinal fluid from a leukemia patient with tubulointerstitial nephritis and meningoencephalitis SO AMERICAN JOURNAL OF KIDNEY DISEASES LA English DT Article DE BK virus (BKV); nephritis; leukemia; meningoencephalitis; virology; cerebrospinal fluid (CSF); urine ID PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY; RENAL-ALLOGRAFT RECIPIENTS; BONE-MARROW TRANSPLANTS; HUMAN POLYOMA-VIRUS; JC-VIRUS; INTERSTITIAL NEPHRITIS; NUCLEOTIDE-SEQUENCE; INFECTION; DISEASE; URINE AB BK virus (BKV) was recovered by polymerase chain reaction (PCR) from brain, kidney, lung, urine, and cerebrospinal fluid (CSF) of a fatal case of BKV tubulointerstitial nephritis with dissemination to lung and brain. Viral regulatory regions in PCR-amplified urine and the lung samples were identical to the archetypal structure, WWT, In the brain and CSF, a rearranged sequence predominated, however. A 94-bp deletion preceded a 71-bp tandem duplication because the same 94-bp segment was deleted from both copies. PCR-amplifled regulatory region products were cloned and sequenced to define further the extent of the rearranged structures. Two kidney clones were archetypal, whereas two others were rearranged differently from the brain and from each other. In contrast to the brain clones, the kidney rearrangements seemed to involve deletion after duplication. Three of four brain clones sequenced were identical to the rearrangement found to dominate in the PCR product. A fourth clone showed two short deletions without any duplication. The four CSF clones all showed rearrangements Identical to that which was amplified by PCR from CSF and brain. This represents the first molecular analysis of a BKV strain obtained from a central nervous system infection, and it reveals regulatory region rearrangements reminiscent of those described in JC virus from brains with progressive multifocal leukoencephalopathy. We suggest that the presence in the CSF of BKV with a dominant rearranged regulatory region may be useful in the diagnosis of BKV meningoencephalitis secondary to BKV nephritis. (C) 2002 by the National Kidney Foundation, Inc. C1 NINCDS, Neurotoxicol Sect, NIH, Bethesda, MD 20892 USA. Yale Univ, Sch Med, Dept Lab Med, New Haven, CT USA. RP Stoner, GL (reprint author), NINCDS, Neurotoxicol Sect, NIH, 36 Convent Dr,Room 4A-27,MSC-4126, Bethesda, MD 20892 USA. NR 44 TC 44 Z9 46 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 0272-6386 J9 AM J KIDNEY DIS JI Am. J. Kidney Dis. PD MAY PY 2002 VL 39 IS 5 BP 1102 EP 1112 DI 10.1053/ajkd.2002.32795 PG 11 WC Urology & Nephrology SC Urology & Nephrology GA 548JQ UT WOS:000175385200023 PM 11979356 ER PT J AU Moawad, AH Goldenberg, RL Mercer, B Meis, PJ Iams, JD Das, A Caritis, SN Miodovnik, M Menard, MK Thurnau, GR Dombrowski, M Roberts, JM AF Moawad, AH Goldenberg, RL Mercer, B Meis, PJ Iams, JD Das, A Caritis, SN Miodovnik, M Menard, MK Thurnau, GR Dombrowski, M Roberts, JM CA NICHD MFMU Network TI The Preterm Prediction Study: The value of serum alkaline phosphatase, alpha-fetoprotein, plasma corticotropin-releasing hormone, and other serum markers for the prediction of spontaneous preterm birth SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE serum alkaline phosphatase; alpha-fetoprotein; corticotropin-releasing hormone; spontaneous preterm birth ID PREGNANCIES; DELIVERY; PLACENTA; LENGTH; RISK AB OBJECTIVE: High levels of a number of analytes are found in maternal blood; alkaline phosphatase, alpha-fetoprotein, and corticotropin-releasing hormone have been associated with spontaneous preterm birth. We investigated the relationship between 8 potential blood markers and subsequent spontaneous preterm birth in asymptomatic pregnant women. STUDY DESIGN: We performed a nested case control study that involved 127 women who were enrolled in the preterm prediction study and who had a singleton spontaneous preterm birth at <35 weeks and 127 women who had a term birth and served as matched (age, parity, center) controls. Serum that was collected at 24 and 28 weeks was analyzed for alkaline phosphatase, a-fetoprotein, corticotropin-releasing hormone, and 5 other analytes. RESULTS: Alkaline phosphatase, a-fetoprotein, and corticotropin-releasing hormone, but not other analytes, were significantly elevated in pregnancies that ended in spontaneous preterm birth. For alkaline phosphatese at 24 weeks, the odds ratio for spontaneous preterm birth at <32 weeks was 6.8 (range, 1.4-32.8) and for spontaneous preterm birth at <35 weeks 5.1 (range, 1.7-15.6). Similar results were found at 28 weeks. For alpha-fetoprotein at 24 weeks, the odds ratio for spontaneous preterm birth at <32 weeks was 8.3 (range, 2.2-30.9) and for spontaneous preterm birth at <35 weeks was 3.5 (range, 1.8-6.7). The levels at 28 weeks were still predictive but less so than at 24 weeks. Corticotropin-re leasing hormone, at 28 weeks but not at 24 weeks, was predictive for spontaneous preterm birth at <35 weeks, with an odds ratio 3.4 (range, 1.0-10.9). CONCLUSION: Elevated alkaline phosphatase and alpha-fetoprotein are associated with subsequent spontaneous preterm birth in asymptomatic pregnant women at 24 and 28 weeks. Elevated corticotropin-releasing hormone levels at 28 weeks are associated with spontaneous preterm birth at <35 weeks. C1 NICHHD, Maternal Fetal Med Unit Network, Bethesda, MD USA. RP Moawad, AH (reprint author), Univ Chicago, Dept Obstet & Gynecol, 5841 S Maryland Ave,MC 2050, Chicago, IL 60637 USA. OI caritis, steve/0000-0002-2169-0712 FU NICHD NIH HHS [HD21414, HD19897, HD21410, HD21434, HD27860, HD27861, HD27869, HD27883, HD27889, HD27905, HD27915, HD27917] NR 31 TC 45 Z9 51 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD MAY PY 2002 VL 186 IS 5 BP 990 EP 996 DI 10.1067/mob.2002.121727 PG 7 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 554WW UT WOS:000175761000033 PM 12015526 ER PT J AU England, LJ Levine, RJ Qian, C Morris, CD Sibai, BM Catalano, PM Curet, LB Klebanoff, MA AF England, LJ Levine, RJ Qian, C Morris, CD Sibai, BM Catalano, PM Curet, LB Klebanoff, MA TI Smoking before pregnancy and risk of gestational hypertension and preeclampsia SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE smoking; gestational hypertension; preeclampsia ID CIGARETTE-SMOKING; NULLIPAROUS WOMEN; THROMBOXANE; PROSTACYCLIN; COHORT; METABOLITES; EXCRETION; CESSATION; NICOTINE; OUTCOMES AB OBJECTIVE: Our purpose was to determine whether smoking before pregnancy reduces the risk of gestational hypertension or preeclampsia. STUDY DESIGN: The trial of Calcium for Preeclampsia Prevention (CPEP) was a randomized study of 4589 nulliparous women conducted in 5 US medical centers during the years 1992 through 1995. Women were classified into 4 exposure groups by smoking history obtained at study enrollment (13-21 weeks' gestation): (1) never smoked, (2) smoked but quit before the last menstrual period (LMP), (3) smoked but quit after LMP but before enrollment, and (4) smoked and still smoking at enrollment. RESULTS: After adjustments were made for maternal age, race, body mass index, type of health insurance, and clinical center, women smoking at enrollment had a reduced risk of hypertension (relative risk = 0.8; 95% CI, 0.6-0.9). Women who quit smoking before the LMP did not have reduced risk (relative risk = 1.1; 95% CI, 0.9-1.3). Results were similar for gestational hypertension and preeclampsia examined separately. CONCLUSION: Women who smoke but quit before becoming pregnant do not have a reduced risk for gestational hypertension or preeclampsia. C1 NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. Oregon Hlth Sci Univ, Dept Med, Portland, OR 97201 USA. Univ Cincinnati, Coll Med, Dept Obstet & Gynecol, Cincinnati, OH 45267 USA. Univ New Mexico, Hlth Sci Ctr, Dept Obstet & Gynecol, Albuquerque, NM 87131 USA. RP England, LJ (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, 6100 Execut Blvd,Room 7B03, Bethesda, MD 20892 USA. FU NICHD NIH HHS [N01-HD-1-3121, N01-HD-1-3122, N01-HD-1-3123, N01-HD-1-3124, N01-HD-1-3125, N01-HD-1-3126, N01-HD-2-3154, N01-HD-5-3246] NR 25 TC 52 Z9 54 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD MAY PY 2002 VL 186 IS 5 BP 1035 EP 1040 DI 10.1067/mob.2002.122404 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 554WW UT WOS:000175761000040 PM 12015533 ER PT J AU Goldenberg, RL Klebanoff, MA Carey, JC AF Goldenberg, RL Klebanoff, MA Carey, JC TI Antimicrobials in the prevention of prematurity - Reply SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Letter C1 NICHHD, Bethesda, MD 20892 USA. Univ Oklahoma, Oklahoma City, OK 73106 USA. RP Goldenberg, RL (reprint author), 1530 3rd Ave S,CIRC 320, Birmingham, AL 35294 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD MAY PY 2002 VL 186 IS 5 BP 1107 EP 1107 DI 10.1067/mob.2002.122979 PG 1 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 554WW UT WOS:000175761000057 ER PT J AU Levine, RJ England, LJ Sibai, BM AF Levine, RJ England, LJ Sibai, BM TI Elevated plasma homocysteine in early pregnancy: A risk factor for the development of severe preeclampsia SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Letter C1 NICHHD, Div Epidemiol Stat & Prevent Res, NIH, Bethesda, MD 20892 USA. Univ Cincinnati, Coll Med, Dept Obstet & Gynecol, Cincinnati, OH 45267 USA. RP Levine, RJ (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, NIH, 6100 Bldg,Room 7B03 MSC 7510, Bethesda, MD 20892 USA. NR 3 TC 2 Z9 2 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD MAY PY 2002 VL 186 IS 5 BP 1107 EP 1107 DI 10.1067/mob.2002.122977 PG 1 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 554WW UT WOS:000175761000058 PM 12015548 ER PT J AU Kolker, DE Olson, SL Dutton-Boilek, J Bennett, KM Wallen, EP Horton, TH Turek, FW AF Kolker, DE Olson, SL Dutton-Boilek, J Bennett, KM Wallen, EP Horton, TH Turek, FW TI Feeding melatonin enhances the phase shifting response to triazolam in both young and old golden hamsters SO AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY LA English DT Article DE aging; circadian ID MAMMALIAN CIRCADIAN CLOCK; SUPRACHIASMATIC NUCLEUS NEURONS; FIRING-RATE RHYTHMS; LIGHT-DARK CYCLE; SYRIAN-HAMSTERS; SIBERIAN HAMSTER; RUNNING ACTIVITY; PERIOD HOMOLOGS; CONSTANT LIGHT; MESSENGER-RNA AB Aging alters many aspects of circadian rhythmicity, including responsivity to phase-shifting stimuli and the amplitude of the rhythm of melatonin secretion. As melatonin is both an output from and an input to the circadian clock, we hypothesized that the decreased melatonin levels exhibited by old hamsters may adversely impact the circadian system as a whole. We enhanced the diurnal rhythm of melatonin by feeding melatonin to young and old hamsters. Animals of both age groups on the melatonin diet showed larger phase shifts than control-fed animals in response to an injection with the benzodiazepine triazolam at a circadian time known to induce phase advances in the activity rhythm of young animals. Thus melatonin treatment can increase the sensitivity of the circadian timing system of young animals to a nonphotic stimulus, and the ability to increase this sensitivity persists into old age, indicating exogenous melatonin might be useful in reversing at least some age-related changes in circadian clock function. C1 Northwestern Univ, Dept Neurobiol & Physiol, Evanston, IL 60208 USA. Northwestern Univ, Ctr Sleep & Circadian Biol, Evanston, IL 60208 USA. Univ Wisconsin Parkside, Dept Biol Sci, Kenosha, WI 53141 USA. RP Turek, FW (reprint author), NICHHD, Endocrinol & Reprod Res Branch, NIH, 49 Convent Dr,Bldg 49,Rm 6A-15,MSC 4510, Bethesda, MD 20892 USA. FU NIA NIH HHS [P01-AG-11412]; NICHD NIH HHS [R01-HD-09885] NR 50 TC 8 Z9 10 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6119 J9 AM J PHYSIOL-REG I JI Am. J. Physiol.-Regul. Integr. Comp. Physiol. PD MAY PY 2002 VL 282 IS 5 BP R1382 EP R1388 DI 10.1152/ajpregu.00362.2001 PG 7 WC Physiology SC Physiology GA 541MC UT WOS:000174987500018 PM 11959680 ER PT J AU Stengel, PW Yamada, M Wess, J Cohen, ML AF Stengel, PW Yamada, M Wess, J Cohen, ML TI M-3-receptor knockout mice: muscarinic receptor function in atria, stomach fundus, urinary bladder, and trachea SO AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY LA English DT Article DE atrial rate; negative chronotropy; smooth muscle contraction; carbamylcholine-mediated responses ID SMOOTH-MUSCLE; M-3 RECEPTOR; RAT STOMACH; SUBTYPES; GENE; STIMULATION; ANTAGONIST; EXPRESSION; RELAXATION; HEART AB Negative chronotropic and smooth muscle contractile responses to the nonselective muscarinic agonist carbamylcholine were compared in isolated tissues from M-3-muscarinic receptor knockout and wild-type mice. Carbamylcholine (10(-8)-3.0 x 10(-5) M) induced a concentration-dependent decrease in atrial rate that was similar in atria from M-3-receptor knockout and wild-type mice, indicating that M-3 receptors were not involved in muscarinic receptor-mediated atrial rate decreases. In contrast, the M-3 receptor was a major muscarinic receptor involved in smooth muscle contraction of stomach fundus, urinary bladder, and trachea, although differences existed in the extent of M-3-receptor involvement among the tissues. Contraction to carbamylcholine was virtually abolished in urinary bladder from M-3-receptor knockout mice, suggesting that contraction was predominantly due to M-3-receptor activation. However, similar to50-60% maximal contraction to carbamylcholine occurred in stomach fundus and trachea from M-3-receptor knockout mice, indicating that contraction in these tissues was also due to M-2- receptor activation. High concentrations of carbamylcholine relaxed the stomach fundus from M-3-receptor knockout mice by M-1-receptor activation. Thus M-3-receptor knockout mice provided unambiguous evidence that M-3 receptors 1) play no role in carbamylcholine-induced atrial rate reduction, 2) are the predominant receptor mediating carbamylcholine-induced urinary bladder contractility, and 3) share contractile responsibility with M-2 receptors in mouse stomach fundus and trachea. C1 Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs, Neurosci Res, Indianapolis, IN 46285 USA. NIDDKD, Bioorgan Chem Lab, Bethesda, MD 20892 USA. RP Stengel, PW (reprint author), Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs, Neurosci Res, Indianapolis, IN 46285 USA. NR 38 TC 88 Z9 96 U1 0 U2 3 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6119 J9 AM J PHYSIOL-REG I JI Am. J. Physiol.-Regul. Integr. Comp. Physiol. PD MAY PY 2002 VL 282 IS 5 BP R1443 EP R1449 DI 10.1152/ajpregu.00486.2001 PG 7 WC Physiology SC Physiology GA 541MC UT WOS:000174987500026 PM 11959688 ER PT J AU Villalobos, ARA Miller, DS Renfro, JL AF Villalobos, ARA Miller, DS Renfro, JL TI Transepithelial organic anion transport by shark choroid plexus SO AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY LA English DT Article DE blood-cerebrospinal fluid barrier; xenobiotic; 2,4-dichlorophenoxyacetic acid; dogfish; elasmobranch ID 2,4-DICHLOROPHENOXYACETIC ACID; 5-HYDROXYINDOLEACETIC ACID; CEREBROSPINAL-FLUID; RAT-KIDNEY; ACCUMULATION; RABBIT; MECHANISM; INVITRO AB Spiny dogfish shark (Squalus acanthias) lateral and IV choroid plexuses (CPs) are ultrastructurally similar to the corresponding tissues of rat. However, shark IV CP is proportionally larger and easily accessible. Moreover, this epithelial sheet can be halved and studied in Ussing flux chambers. We have used confocal fluorescence microscopy and radiotracer techniques to characterize transepithelial transport of the organic anions (OAs) fluorescein (FL) and 2,4-dichlorophenoxyacetic acid (2,4-D), respectively, by shark CP. Lateral and IV CP accumulated 1 muM FL, with highest levels in the underlying extracellular spaces, intermediate levels in epithelial cells, and lowest levels in the medium. 2,4-D and probenecid inhibited FL accumulation in cells and extracellular spaces, suggesting that these substrates compete for common carriers. Unidirectional absorptive [cerebrospinal fluid (CSF)-to-blood] and secretory (blood-to-CSF) fluxes of 10 muM [C-14]2,4-D were measured under short-circuited conditions in IV CP mounted in Ussing chambers. 2,4-D underwent net absorption, with an average flux ratio of 7. Probenecid, 2,4,5-trichlorophenoxyacetic acid, and 5-hydroxyindolacetic acid reduced net absorption, reversibly inhibiting unidirectional absorption, with no effect on secretion. Ouabain irreversibly reduced net 2,4-D absorption and cellular and extracellular accumulation of FL, suggesting energetic coupling of OA absorption to Na+ transport. Collectively, these data indicate that shark CP actively removes OAs from CSF by a process that is specific and active. C1 Univ Rochester, Dept Environm Med, Rochester, NY 14642 USA. Mt Desert Isl Biol Lab, Salsbury Cove, ME 04672 USA. NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. Univ Connecticut, Dept Physiol & Neurobiol, Storrs, CT 06269 USA. RP Villalobos, ARA (reprint author), Univ Rochester, Dept Environm Med, 575 Elmwood Ave,Box EHSC, Rochester, NY 14642 USA. FU NIEHS NIH HHS [P30 ES01247, P30 ES 03828, ES-10439]; NINDS NIH HHS [NS39452] NR 31 TC 14 Z9 14 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6119 J9 AM J PHYSIOL-REG I JI Am. J. Physiol.-Regul. Integr. Comp. Physiol. PD MAY PY 2002 VL 282 IS 5 BP R1308 EP R1316 DI 10.1152/ajpregu.00677.2001 PG 9 WC Physiology SC Physiology GA 541MC UT WOS:000174987500008 PM 11959670 ER PT J AU Breen, CM Sykes, DB Fricker, G Miller, DS AF Breen, CM Sykes, DB Fricker, G Miller, DS TI Confocal imaging of organic anion transport in intact rat choroid plexus SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE transepithelial transport; potential-driven efflux; blood-brain barrier; 2,4-dichlorophenoxyacetic acid; fluorescein ID CEREBROSPINAL FLUID BARRIER; RESISTANCE; SECRETION; BRAIN; CELLS; BENZYLPENICILLIN; ACCUMULATION; MICROSCOPY; MEMBRANE; PROTEIN AB We used confocal microscopy and quantitative image analysis to follow the movement of the fluorescent organic anion fluorescein (FL) from bath to cell and cell to blood vessel in intact rat lateral choroid plexus. FL accumulation in epithelial cells and underlying vessels was rapid, concentrative, and reduced by other organic anions. At steady state, cell fluorescence exceeded bath fluorescence by a factor of 3-5, and vessel fluorescence exceeded cell fluorescence by a factor of similar to2. In cells, FL distributed between diffuse and punctate compartments. Cell and vessel accumulation of FL decreased when metabolism was inhibited by KCN, when bath Na(+) was reduced from 130 to 26 mM, and when the Na(+) gradient was collapsed with ouabain. Cell and vessel accumulation increased by >50% when 1-10 muM glutarate was added to the bath. Finally, transport of FL and carboxyfluorescein (generated intracellularly from carboxyfluorescein diacetate) from cell to blood vessel was greatly diminished when medium K(+) concentration ([K(+)]) was increased 10-fold. These results 1) validate a new approach to the study of choroid plexus function, and 2) indicate a two-step mechanism for transepithelial organic anion transport: indirect coupling of uptake to Na(+) at the apical membrane and electrical potential-driven efflux at the basolateral membrane. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. Inst Pharmazeut Technol & Biopharm, INF 366, D-69120 Heidelberg, Germany. RP Miller, DS (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. EM miller@niehs.nih.gov NR 28 TC 22 Z9 22 U1 1 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1931-857X J9 AM J PHYSIOL-RENAL JI Am. J. Physiol.-Renal Physiol. PD MAY PY 2002 VL 282 IS 5 BP F877 EP F885 DI 10.1152/ajprenal.00171.2001 PG 9 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 537VA UT WOS:000174779200013 PM 11934698 ER PT J AU Miyaji, T Hu, XZ Star, RA AF Miyaji, T Hu, XZ Star, RA TI alpha-Melanocyte-simulating hormone and interleukin-10 do not protect the kidney against mercuric chloride-induced injury SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE mercury; acute renal failure; leukocytes ID ACUTE-RENAL-FAILURE; ISCHEMIA-REPERFUSION INJURY; STIMULATING HORMONE; INORGANIC MERCURY; NITRIC-OXIDE; MICE; RAT; ICAM-1; ISCHAEMIA/REPERFUSION; NEPHROTOXICITY AB The anti-inflammatory cytokines alpha-melanocyte-stimulating hormone (MSH) and interleukin (IL)-10 inhibit acute renal failure (ARF) after ischemia or cisplatin administration; however, these agents have not been tested in a pure nephrotoxic model of ARF. Therefore, we examined the effects of alpha-MSH and IL-10 in HgCl(2)-induced ARF. Mice were injected subcutaneously with HgCl(2) and then given vehicle, alpha-MSH, or IL-10 by intravenous injection. Animals were killed to study serum creatinine, histology, and myeloperoxidase activity. Treatment with either alpha-MSH or IL-10 did not alter the increase in serum creatinine, tubular damage, or leukocyte accumulation at 48 h after HgCl(2) injection. Because alpha-MSH and IL-10 are active in other injury models that involve leukocytes, we studied the time course of tubular damage and leukocyte accumulation to investigate whether leukocytes caused the tubular damage or accumulated in response to the tubular damage. Tubular damage was present in the outer stripe 12 h after HgCl(2) injection. In contrast, the number of leukocytes and renal myleoperoxidase activity were normal at 12 h but were significantly increased at 24 and 48 h after injection. We conclude that neither alpha-MSH nor IL-10 altered the course of HgCl(2)-induced renal injury. Because the tubular damage preceded leukocyte infiltration, the delayed leukocyte accumulation may play a role in the removal of necrotic tissue and/or tissue repair in HgCl(2)- induced ARF. C1 NIDDKD, Renal Diagnost & Therapeut Unit, NIH, Bethesda, MD 20892 USA. RP Star, RA (reprint author), NIDDKD, Renal Diagnost & Therapeut Unit, NIH, Rm 3N108,10 Ctr Dr, Bethesda, MD 20892 USA. EM Robert_Star@nih.gov NR 41 TC 15 Z9 15 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1931-857X J9 AM J PHYSIOL-RENAL JI Am. J. Physiol.-Renal Physiol. PD MAY PY 2002 VL 282 IS 5 BP F795 EP F801 DI 10.1152/ajprenal.00203.2001 PG 7 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 537VA UT WOS:000174779200003 PM 11934688 ER PT J AU Norris, SL Nichols, PJ Caspersen, CJ Glasgow, RE Engelgau, MM Jack, L Isham, G Snyder, SR Carande-Kulis, VG Garfield, S Briss, P McCulloch, D AF Norris, SL Nichols, PJ Caspersen, CJ Glasgow, RE Engelgau, MM Jack, L Isham, G Snyder, SR Carande-Kulis, VG Garfield, S Briss, P McCulloch, D CA Task Force Community Preventive Se TI The effectiveness of disease and case management for people with diabetes - A systematic review SO AMERICAN JOURNAL OF PREVENTIVE MEDICINE LA English DT Review DE community health services; diabetes mellitus; evidence-based medicine; preventive health services; public health practice; review literature ID HEALTH MAINTENANCE ORGANIZATION; LOWER-EXTREMITY AMPUTATION; INTENSIVE INSULIN THERAPY; IMPROVED GLYCEMIC CONTROL; CONGESTIVE-HEART-FAILURE; NURSE CASE-MANAGEMENT; FOLLOW-UP; PRIMARY-CARE; RANDOMIZED TRIAL; WEIGHT-LOSS AB This report presents the results of a systematic review of the effectiveness and economic efficiency of disease management and case management for people with diabetes and forms the basis for recommendations by the Task Force on Community Preventive Services on the use of these two interventions. Evidence supports the effectiveness of disease management on glycemic control; on screening for diabetic retinopathy, foot lesions and peripheral neuropathy, and proteinuria; and on the monitoring of lipid concentrations. This evidence is applicable to adults with diabetes in managed care organizations and community clinics in the United States and Europe. Case management is effective in improving both glycemic control and provider monitoring of glycemic control. This evidence is applicable primarily in the U.S. managed care setting for adults with type 2 diabetes. Case management is effective both when delivered in conjunction with disease management and when delivered with one or more additional educational, reminder, or support interventions. C1 CDCP, Div Diabet Translat, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA 30341 USA. CDCP, Epidemiol Program Off, Atlanta, GA 30341 USA. AMC Canc Res Ctr, Denver, CO USA. HealthPartners, Minneapolis, MN USA. NIDDKD, Diabet Program Branch, NIH, Bethesda, MD 20892 USA. Grp Hlth Cooperat Puget Sound, Seattle, WA 98121 USA. RP Norris, SL (reprint author), CDCP, Div Diabet Translat, Natl Ctr Chron Dis Prevent & Hlth Promot, MS K-10,4770 Buford Highway NE, Atlanta, GA 30341 USA. RI Caspersen, Carl/B-2494-2009 NR 157 TC 237 Z9 247 U1 9 U2 28 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0749-3797 J9 AM J PREV MED JI Am. J. Prev. Med. PD MAY PY 2002 VL 22 IS 4 SU S BP 15 EP 38 DI 10.1016/S0749-3797(02)00423-3 PG 24 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 550JE UT WOS:000175499200006 PM 11985933 ER PT J AU Norris, SL Nichols, PJ Caspersen, CJ Glasgow, RE Engelgau, MM Jack, L Snyder, SR Carande-Kulis, VG Isham, G Garfield, S Briss, P McCulloch, D AF Norris, SL Nichols, PJ Caspersen, CJ Glasgow, RE Engelgau, MM Jack, L Snyder, SR Carande-Kulis, VG Isham, G Garfield, S Briss, P McCulloch, D CA Task Force Commun Prevent Serv TI Increasing diabetes self-management education in community settings - A systematic review SO AMERICAN JOURNAL OF PREVENTIVE MEDICINE LA English DT Review DE blood glucose self-monitoring; community health services; decision making; diabetes mellitus; evidence-based medicine; health education; patient education; preventive health services; public health practice; review literature; self-care; self-efficacy; self-help groups ID IMPAIRED GLUCOSE-TOLERANCE; INTENSIVE INSULIN THERAPY; GLYCEMIC CONTROL; PATIENT EDUCATION; CONTROLLED TRIAL; WEIGHT-LOSS; ANCHORED INSTRUCTION; MEXICAN-AMERICANS; META-ANALYSIS; FOLLOW-UP AB This report presents the results of a systematic review of the effectiveness and economic efficiency of self-management education interventions for people with diabetes and forms the basis for recommendations by the Task Force on Community Preventive Services. Data on glycemic control provide sufficient evidence that self-management education is effective in community gathering places for adults with type 2 diabetes and in the home for adolescents with type 1 diabetes. Evidence is insufficient to assess the effectiveness of self-management education interventions at the worksite or in summer camps for either type 1 or type 2 diabetes or in the home for type 2 diabetes. Evidence is also insufficient to assess the effectiveness of educating coworkers and school personnel about diabetes. C1 CDCP, Div Diabet Translat, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA 30341 USA. CDCP, Epidemiol Program Off, Atlanta, GA 30341 USA. AMC Canc Res Ctr, Denver, CO USA. HealthPartners, Minneapolis, MN USA. NIDDKD, Diabet Program Branch, Bethesda, MD 20892 USA. Grp Hlth Cooperat Puget Sound, Seattle, WA 98121 USA. RP Norris, SL (reprint author), CDCP, Div Diabet Translat, Natl Ctr Chron Dis Prevent & Hlth Promot, MS K-10,4770 Buford Highway NE, Atlanta, GA 30341 USA. RI Caspersen, Carl/B-2494-2009 NR 124 TC 154 Z9 155 U1 1 U2 18 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0749-3797 J9 AM J PREV MED JI Am. J. Prev. Med. PD MAY PY 2002 VL 22 IS 4 SU S BP 39 EP 66 DI 10.1016/S0749-3797(02)00424-5 PG 28 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 550JE UT WOS:000175499200007 PM 11985934 ER PT J AU Kahn, EB Ramsey, LT Brownson, RC Heath, GW Howze, EH Powell, KE Stone, EJ Rajab, MW Corso, P Briss, PA AF Kahn, EB Ramsey, LT Brownson, RC Heath, GW Howze, EH Powell, KE Stone, EJ Rajab, MW Corso, P Briss, PA CA Task Force Commun Prevent Serv TI The effectiveness of interventions to increase physical activity - A systematic review SO AMERICAN JOURNAL OF PREVENTIVE MEDICINE LA English DT Review ID ELEMENTARY-SCHOOL-CHILDREN; CARDIOVASCULAR-DISEASE PREVENTION; DEPENDENT DIABETES-MELLITUS; CORONARY HEART-DISEASE; BONE-MINERAL DENSITY; WEIGHT-LOSS PROGRAM; HEALTH PROMOTION PROGRAM; HARVARD ALUMNI HEALTH; RISK-FACTORS; LIFE-STYLE AB The Guide to Community Preventive Service's methods for systematic reviews were used to evaluate the effectiveness of various approaches to increasing physical activity: informational, behavioral and social, and environmental and policy approaches. Changes in physical activity behavior and aerobic capacity were used to assess effectiveness. Two informational interventions ("point-of-decision" prompts to encourage stair use and community-wide campaigns) were effective, as were three behavioral and social interventions (school-based physical education, social support in community settings, and individually-adapted health behavior change) and one environmental and policy intervention (creation of or enhanced access to places for physical activity combined with informational outreach activities). Additional information about applicability, other effects, and barriers to implementation are provided for these interventions. Evidence is insufficient to assess a number of interventions: classroom-based health education focused on information provision, and family-based social support (because of inconsistent findings); mass media campaigns and college-based health education and physical education (because of an insufficient number of studies); and classroom-based health education focused on reducing television viewing and video game playing (because of insufficient evidence of an increase in physical activity). These recommendations should serve the needs of researchers, planners, and other public health decision makers. C1 CDCP, Div Prevent Res & Analyt Methods, Epidemiol Program Off, Atlanta, GA 30341 USA. CDCP, Div Nutr & Phys Act, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA 30341 USA. Task Force Community Prevent Serv, St Louis, MO USA. St Louis Univ, Sch Publ Hlth, St Louis, MO 63103 USA. Agcy Tox Subst & Dis Registry, Div Hlth Educ & Promot, Atlanta, GA USA. Georgia Dept Human Resources, Epidemiol & Prevent Branch, Atlanta, GA USA. NHLBI, NIH, Bethesda, MD 20892 USA. RP Briss, PA (reprint author), CDCP, Community Guide Branch, 4770 Buford Highway,MS-K73, Atlanta, GA 30341 USA. OI Kahn, Emily/0000-0001-7812-7958 NR 160 TC 930 Z9 949 U1 47 U2 342 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0749-3797 J9 AM J PREV MED JI Am. J. Prev. Med. PD MAY PY 2002 VL 22 IS 4 SU S BP 73 EP 108 DI 10.1016/S0749-3797(02)00434-8 PG 36 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 550JE UT WOS:000175499200009 PM 11985936 ER PT J AU Rimer, BK Halabi, S Skinner, CS Lipkus, IM Strigo, TS Kaplan, EB Samsa, GP AF Rimer, BK Halabi, S Skinner, CS Lipkus, IM Strigo, TS Kaplan, EB Samsa, GP TI Effects of a mammography decision-making intervention at 12 and 24 months SO AMERICAN JOURNAL OF PREVENTIVE MEDICINE LA English DT Article DE counseling; decision making; decision support techniques; mammography; telephone ID BREAST-CANCER RISK; INCREASE MAMMOGRAPHY; SCREENING MAMMOGRAPHY; FUTURE-DIRECTIONS; SMOKING CESSATION; FAMILY HISTORY; WOMEN; COMMUNICATION; BEHAVIOR; ADOPTION AB Background: Most women are not getting regular mammograms, and there is confusion about several mammography-related issues, including the age at which women should begin screening. Numerous groups have called for informed decision making about mammography, but few programs have resulted. Our research is intended to fill this gap. Methods: We conducted a randomized controlled trial, which ran from 1997 to 2000. Women aged 40 to 44 and 50 to 54, who were enrolled in Blue Cross Blue Shield of North Carolina, were randomly assigned to one of three groups: usual care (UC), tailored print (TP) materials, or TP plus tailored telephone counseling (TP+TC). We assessed the impact of tailored interventions on knowledge about breast cancer and mammography, accuracy of breast cancer risk perceptions, and use of mammography at two time points after intervention-12 and 24 months. Results: At 12 and 24 months, women who received TP+TC had significantly greater knowledge and more accurate breast cancer risk perceptions. Compared to UC, they were 40% more likely to have had mammograms (odds ratio=0.9-2.1). The effect was primarily for women in their 50s. TP had significant effects for knowledge and accuracy, but women who received TP were less likely to have had mammography. Conclusions: Decision-making interventions, comprised of two tailored print interventions (booklet and newsletter), delivered a year apart, with or without two tailored telephone calls, significantly increased knowledge and accuracy of perceived breast cancer risk at 12 and 24 months post-intervention. The effect on mammography use was significant in bivariate relationships but had a much more modest impact in multivariate analyses. C1 NCI, Div Canc Control & Populat Sci, Rockville, MD 20852 USA. Duke Comprehens Canc Ctr, Dept Biostat & Bioinformat, Canc Prevent Detect & Control Res Program, Durham, NC USA. Duke Univ, Med Ctr, Ctr Canc, Durham, NC USA. RP Rimer, BK (reprint author), NCI, Div Canc Control & Populat Sci, 6130 Execut Blvd,Room 6138, Rockville, MD 20852 USA. FU NCI NIH HHS [5P01-CA-72099-03] NR 65 TC 83 Z9 84 U1 3 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0749-3797 J9 AM J PREV MED JI Am. J. Prev. Med. PD MAY PY 2002 VL 22 IS 4 BP 247 EP 257 AR PII S0749-3797(02)00417-8 DI 10.1016/S0749-3797(02)00417-8 PG 11 WC Public, Environmental & Occupational Health; Medicine, General & Internal SC Public, Environmental & Occupational Health; General & Internal Medicine GA 549EH UT WOS:000175431200005 PM 11988381 ER PT J AU Alexander, GE Chen, K Pietrini, P Rapoport, SI Reiman, EM AF Alexander, GE Chen, K Pietrini, P Rapoport, SI Reiman, EM TI Longitudinal PET evaluation of cerebral metabolic decline in dementia: A potential outcome measure in Alzheimer's disease treatment studies SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article; Proceedings Paper CT World Alzheimer Congress 2000 CY JUL 09-18, 2000 CL WASHINGTON, D.C. ID POSITRON-EMISSION-TOMOGRAPHY; COGNITIVE DECLINE; GLUCOSE-UTILIZATION; APOLIPOPROTEIN-E; RISK AB Objective: It is well established that regional cerebral metabolic rates for glucose assessed by [F-18]fluorodeoxyglucose (FDG) positron emission tomography (PET) in patients with Alzheimer's disease in the mental resting state (eyes and ears covered) provide a sensitive, in vivo metabolic index of Alzheimer's disease dementia. Few studies, however, have evaluated longitudinal declines in regional cerebral glucose metabolism in patients with dementia caused by Alzheimer's disease, in addition, the available studies have not used recently developed brain mapping algorithms to characterize the progression of Alzheimer's disease throughout the brain, and none considered the statistical power of regional cerebral glucose metabolism in testing the ability of treatments to attenuate the progression of dementia. Method: The authors used FDG PET and a brain mapping algorithm to investigate cross-sectional reductions in regional cerebral glucose metabolism, longitudinal decline in regional cerebral glucose metabolism after a 1-year follow-up, and the power of this method to evaluate treatments for Alzheimer's disease in patients with mild to moderate dementia. PET scans were initially acquired in 14 patients with Alzheimer's disease and 34 healthy comparison subjects of similar age and sex. Repeat scans were obtained in the patients 1 year later. Power analyses for voxels showing maximal decline over the 1-year period in regional cerebral glucose metabolism (mg/100 g per minute) were computed to estimate the sample sizes needed to detect a significant treatment response in a 1-year, double-blind, placebo-controlled treatment study. Results: The patients with Alzheimer's disease had significantly lower glucose metabolism than healthy comparison subjects in parietal, temporal, occipital, frontal, and posterior cingulate cortices, One year later, the patients, with Alzheimer's disease had significant declines in glucose metabolism in parietal, temporal, frontal, and posterior cingulate cortices. Using maximal glucose metabolism reductions in the left frontal cortex, we estimated that as few as 36 patients per group would be needed to detect a 33% treatment response with one-tailed significance of pless than or equal to0.005 and 80% power in a 1-year, double-blind, placebo-controlled treatment study. Conclusions: These findings indicate that brain metabolism as assessed by FDG PET during mental rest is a sensitive marker of disease progression in Alzheimer's disease over a 1-year period. These findings also support the feasibility of using FDG PET as an outcome measure to test the ability of treatments to attenuate the progression of Alzheimer's disease. C1 Arizona State Univ, Dept Psychol, Tempe, AZ 85287 USA. Arizona State Univ, Arizona Alzheimers Res Ctr, Tempe, AZ 85287 USA. Arizona State Univ, Dept Math, Tempe, AZ 85287 USA. Univ Arizona, Dept Psychiat, Tucson, AZ USA. Banner Hlth, Good Samaritan PET Ctr, Phoenix, AZ USA. Univ Pisa, Dept Expt Pathol Med Biotechnol Infectivol & Epid, I-56100 Pisa, Italy. NIA, Brain Physiol & Metab Sect, NIH, Bethesda, MD 20892 USA. RP Alexander, GE (reprint author), Arizona State Univ, Dept Psychol, Tempe, AZ 85287 USA. RI Chen, kewei/P-6304-2015 OI Chen, kewei/0000-0001-8497-3069 NR 33 TC 269 Z9 276 U1 1 U2 10 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD MAY PY 2002 VL 159 IS 5 BP 738 EP 745 DI 10.1176/appi.ajp.159.5.738 PG 8 WC Psychiatry SC Psychiatry GA 548BD UT WOS:000175367400009 PM 11986126 ER PT J AU Fee, E Brown, TM Lazarus, J Theerman, P AF Fee, E Brown, TM Lazarus, J Theerman, P TI Baxter Street then SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Article C1 NIH, Natl Lib Med, Hist Med Div, Bethesda, MD 20892 USA. Univ Rochester, Dept Community & Prevent Med, Rochester, NY USA. Univ Rochester, Dept Hist, Rochester, NY USA. RP Fee, E (reprint author), Bldg 38,Room 1E21,8600 Rockville Pike, Bethesda, MD 20894 USA. NR 3 TC 0 Z9 0 U1 0 U2 0 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD MAY PY 2002 VL 92 IS 5 BP 753 EP 753 DI 10.2105/AJPH.92.5.753 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 545MV UT WOS:000175222200022 PM 11988440 ER PT J AU Fee, E Brown, TM AF Fee, E Brown, TM TI John Henry Bell - Occupational anthrax pioneer SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Biographical-Item C1 NIH, Natl Lib Med, Hist Med Div, Bethesda, MD 20892 USA. RP Fee, E (reprint author), Bldg 38,Room 1E21,8600 Rockville Pike, Bethesda, MD 20894 USA. NR 0 TC 3 Z9 3 U1 0 U2 1 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD MAY PY 2002 VL 92 IS 5 BP 756 EP 757 DI 10.2105/AJPH.92.5.756 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 545MV UT WOS:000175222200024 ER PT J AU Wine, RN Dial, JM Tomer, KB Borchers, CH AF Wine, RN Dial, JM Tomer, KB Borchers, CH TI Identification of components of protein complexes using a fluorescent photo-cross-linker and mass spectrometry SO ANALYTICAL CHEMISTRY LA English DT Article ID RAT SEMINIFEROUS EPITHELIUM; SEQUENCE DATABASES; ALPHA-6-BETA-1; SPERMIATION; INFORMATION; ADHESION; GELS; CD4 AB This study describes a novel method for improving the specific recognition, detection, and identification of proteins involved in multiprotein complexes. The method is based on a combination of coinummoprecipitation, chemical cross-linking, and specific fluorescent tagging of protein components in close association with one another. Specific fluorescent tagging of the protein complex components was achieved using the cleavable, fluorescent cross-linker sulfosuccinimidyl 2-(7-azido-4-methylcoumarin-3-acetamido) ethyl-1,3'-dithiopropionate (SAED). Following dissociation and separation by SDS-PAGE, the fluorescently tagged proteins are then visualized by UV illumination, excised, and, following in-gel digestion, identified by mass spectrometry. In this study, a complex of the HFV-envelope protein gp120 and its cellular receptor CD4 was used as a model system. The sensitivity of detection of fluorescent SAED-labeled proteins in SDS gels, and the sensitivity of the mass spectrometric identification of fluorescent proteins after in-gel digestion, is in the range of a few hundred femtomoles of protein. This sensitivity is comparable to that achieved with silver-staining techniques, but fluorescence detection is protein independent and no background interference occurs. Furthermore, fluorescence labeling is significantly more compatible with mass spectrometric identification of proteins than is silver staining. The first application of this strategy was in the investigation of the mechanism of spermiation, the process by which mature spermatids separate from Sertoli cells. For the coimmunoprecipitation experiment, an antibody against paxillin, a protein involved in spermatid-Sertoli cell junctional complexes, was used. More components of the paxillin protein complex were visible by fluorescence detection of SAED-labeled proteins than were visible on comparable silver-stained gels. Mass spectrometric analysis of the fluorescently labeled proteins identified integrin alpha(6) precursor as a protein associated in a complex with paxillin. The identification of integrin alpha(6) precursor was confirmed by Western blot analysis and verifies the applicability of this novel approach for identifying proteins involved in protein complexes. C1 NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27713 USA. NIEHS, Toxicol Lab, NIH, Res Triangle Pk, NC 27713 USA. RP Borchers, CH (reprint author), Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA. RI Tomer, Kenneth/E-8018-2013 NR 26 TC 32 Z9 34 U1 0 U2 9 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 MAY 1 PY 2002 VL 74 IS 9 BP 1939 EP 1945 DI 10.1021/ac011041w PG 7 WC Chemistry, Analytical SC Chemistry GA 547XU UT WOS:000175358800026 PM 12033289 ER PT J AU Zuzak, KJ Schaeberle, MD Lewis, EN Levin, IW AF Zuzak, KJ Schaeberle, MD Lewis, EN Levin, IW TI Visible reflectance hyperspectral imaging: Characterization of a noninvasive, in vivo system for determining tissue perfusion SO ANALYTICAL CHEMISTRY LA English DT Article ID INFRARED SPECTROSCOPIC ASSESSMENT; OXYGEN-SATURATION; INVIVO; MICROCIRCULATION; FILTERS AB We characterize a visible reflectance hyperspectral imaging system for noninvasive, in vivo, quantitative analysis of human tissue in a clinical environment. The subject area is illuminated with a quartz-tungsten-halogen light source, and the reflected light is spectrally discriminated by a liquid crystal tunable filter (LCTF) and imaged onto a silicon charge-coupled device detector. The LCTF is continuously tunable within its useful visible spectral range (525-725 nm) with an average spectral full width at half-height bandwidth of 0.38 nm and an average transmittance of 10.0%. A standard resolution target placed 5.5 ft from the system results in a field of view with a 17-cm diameter and an optimal spatial resolution of 0.45 mm. The measured reflectance spectra are quantified in terms of apparent absorbance and formatted as a hyperspectral image cube. As a clinical example, we examine a model of vascular dysfunction involving both ischemia and reactive hyperemia during tissue reperfusion. In this model, spectral images, based upon oxyhemoglobin and deoxyhemoblobin signals in the 525-645-nm region, are deconvoluted using a multivariate least-squares regression analysis to visualize the spatial distribution of the percentages of oxyhemoglobin and deoxyhemoglobin in specific skin tissue areas. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Levin, IW (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5,Room B1-32, Bethesda, MD 20892 USA. NR 30 TC 114 Z9 115 U1 0 U2 21 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 MAY 1 PY 2002 VL 74 IS 9 BP 2021 EP 2028 DI 10.1021/ac011275f PG 8 WC Chemistry, Analytical SC Chemistry GA 547XU UT WOS:000175358800039 PM 12033302 ER PT J AU Gilligan, JJ Schuck, P Yergey, AL AF Gilligan, JJ Schuck, P Yergey, AL TI Mass spectrometry after capture and small-volume elution of analyte from a surface plasmon resonance biosensor SO ANALYTICAL CHEMISTRY LA English DT Article ID BIOMOLECULAR INTERACTION ANALYSIS; IDENTIFICATION; IONIZATION; PROTEOMICS; PROTEINS; BIA/MS; MATRIX AB The identification of binding partners of proteins by mass spectrometry following specific capture on a biosensor surface is a promising tool for proteomics research and the identification and characterization of protein-protein interactions. Previous approaches include the direct ionization of analyte from the biosensor chip on a matrix assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOFMS) apparatus and the on-chip digestion followed by elution, chromatographic concentration of the fragments, and electrospray mass spectrometry. In the present paper, using the small-volume microfluidic sample manipulation technique with oscillatory flow reported recently (Abrantes et al. Anal. Chem. 200 1, 73, 2828-2835), analyte is shown to be eluted from the sensor surface into a small volume of buffer that promotes dissociation from the capture surface and delivery to the mass spectrometer. Both the incubation of the sensor surface with the sample and the recovery of analyte can be achieved with a few microliters and conducted until steady-state is attained. Because the procedure is nondestructive for the sensor surface, multiple cycles of capture and elution allow the transfer and concentration of analyte into the elution buffer. The eluted analyte can be studied directly by MALDI-TOFMS, or subjected to proteolytic digestion for protein identification. Transfer into the elution buffer and MALDI-TOFMS detection was achieved from 5 muL of starting samples containing <50 fmol of analyte. Examples are presented for the specific detection and recovery of a protein from a complex mixture of cytosolic proteins. C1 NICHHD, Sect Metab Anal & Mass Spectrometry, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. NIH, Div Bioengn & Phys Sci, ORS, OD, Bethesda, MD 20892 USA. RP Yergey, AL (reprint author), NICHHD, Sect Metab Anal & Mass Spectrometry, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. OI Schuck, Peter/0000-0002-8859-6966 NR 24 TC 38 Z9 39 U1 0 U2 7 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 MAY 1 PY 2002 VL 74 IS 9 BP 2041 EP 2047 DI 10.1021/ac015642z PG 7 WC Chemistry, Analytical SC Chemistry GA 547XU UT WOS:000175358800042 PM 12033305 ER PT J AU Sang, CN Booher, S Gilron, I Parada, S Max, MB AF Sang, CN Booher, S Gilron, I Parada, S Max, MB TI Dextromethorphan and memantine in painful diabetic neuropathy and postherpetic neuralgia - Efficacy and dose-response trials SO ANESTHESIOLOGY LA English DT Article ID ASPARTATE RECEPTOR ANTAGONIST; AMINO-ACID ANTAGONISTS; DOUBLE-BLIND; RAT MODEL; PERIPHERAL NEUROPATHY; THERMAL HYPERALGESIA; NR2B SUBUNIT; DORSAL-HORN; CROSS-OVER; KETAMINE AB Background: There are few repeated dose-controlled trials of N-methyl-D-aspartate glutamate receptor antagonists in patients with neuropathic pain. The authors sought to evaluate two low-affinity N-methyl-D-aspartate antagonists using a novel two-stage design. Methods. The authors studied patients with painful diabetic neuropathy (DN) and postherpetic neuralgia (PHN) in two crossover trials: (1) efficacy trial (dextromethorphan vs. memantine rs. active placebo [lorazepam]) and (2) dose-response trial of the preferred active drug in responders from the first study (0% vs. 25% vs. 50% vs. 100% of each patient's maximally tolerated dose). Pain intensity was measured on a 20-point scale. Results: Nineteen of 23 DN patients and 17 of 21 PHN patients completed the efficacy trial. Median doses for DN and PHN were 400 and 400 mg/day dextromethorplian, 55 and 35 mg/day memantine, and 1.8 and 1.2 mg/day lorazepam. In the efficacy trial, among patients with DN, dextromethorphan reduced pain intensity by a mean of 33% from baseline, memantine reduced pain intensity by a mean of 17%, and lorazepam reduced pain intensity by a mean of 16%; the proportions of subjects achieving greater than moderate pain relief were 68% with dextromethorphan, 47% with memantine, and 37% with lorazepam. Mean reductions in pain intensity in patients with PHN were 6% with dextromethorphan, 2% with memantine, and 0% with lorazepam. No comparison with placebo reached statistical significance in the efficacy trial. In the 10 DN subjects who responded to dextromethorphan, there was a significant dose-response effect on pain Intensity (P = 0.035), with the highest dose significantly better than that of lorazepam (P = 0.03). Conclusions: Dextromethorphan is effective in a dose-related fashion in selected patients with DN. This was not true of PUN, suggesting a difference in pain mechanisms. Selective approaches to pain-relevant N-methyl-D-aspartate receptors are warranted. C1 Massachusetts Gen Hosp, Dept Anesthesia, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Anesthesia, Bethesda, MD USA. Natl Inst Dent & Craniofacial Res, Pain & Neurosensory Mech Branch, Bethesda, MD USA. NIH, Ctr Clin, Dept Nursing, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Dept Anesthesia & Crit Care, Boston, MA USA. RP Sang, CN (reprint author), Massachusetts Gen Hosp, Dept Anesthesia, 55 Fruit St,Clin 3, Boston, MA 02114 USA. EM csang@partners.org FU NIDCR NIH HHS [ZO1-DE00366] NR 45 TC 139 Z9 148 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0003-3022 J9 ANESTHESIOLOGY JI Anesthesiology PD MAY PY 2002 VL 96 IS 5 BP 1053 EP 1061 DI 10.1097/00000542-200205000-00005 PG 9 WC Anesthesiology SC Anesthesiology GA 547EU UT WOS:000175320700004 PM 11981142 ER PT J AU Marchbanks, PA McDonald, JA Wilson, HG Burnett, NM Daling, JR Bernstein, L Malone, KE Strom, BL Norman, SA Weiss, LK Liff, JM Wingo, PA Burkman, RT Folger, SG Berlin, JA Deapen, DM Ursin, G Coates, RJ Simon, MS Press, MF Spirtas, R AF Marchbanks, PA McDonald, JA Wilson, HG Burnett, NM Daling, JR Bernstein, L Malone, KE Strom, BL Norman, SA Weiss, LK Liff, JM Wingo, PA Burkman, RT Folger, SG Berlin, JA Deapen, DM Ursin, G Coates, RJ Simon, MS Press, MF Spirtas, R TI The NICHD women's contraceptive and reproductive experiences study: Methods and operational results SO ANNALS OF EPIDEMIOLOGY LA English DT Article DE oral contraceptives; hormones; breast cancer; epidemiology; case-control studies ID BREAST-CANCER; RISK; DESIGN AB PURPOSE: This paper presents methods and operational results of a population-based case-control study examining the effects of oral contraceptive use on breast cancer risk among white and black women aged 35-64 years in five U.S. locations. METHODS: Cases were women newly diagnosed with breast cancer during July 1994 through April 1998. Controls were identified through random digit dialing (RDD) using unclustered sampling with automated elimination of nonworking numbers. Sampling was density-based, with oversampling of black women. In-person interviews were conducted from August 1994 through December 1998. Blood samples were obtained from subsets of cases and controls, and tissue samples were obtained from subsets of cases. A computerized system tracked subjects through study activities. Special attention was devoted to minimizing exposure misclassification, because any exposure-disease associations were expected to be small. RESULTS: An estimated 82% of households were screened successfully through RDD. Interviews were completed for 4575 cases (2953 whites; 1622 blacks) and 4682 controls (3021 whites; 1661 blacks). Interview response rates for cases and controls were 76.5% and 78.6%, respectively, with lower rates for black women and older women. CONCLUSIONS: The methodologic details of this large collaboration may assist researchers conducting similar investigations. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Ctr Dis Control & Prevent, Div Reprod Hlth MS K34, Atlanta, GA 30333 USA. NICHHD, Contracept & Reprod Hlth Branch, Populat Res Ctr, Bethesda, MD 20892 USA. Wayne State Univ, Karmanos Canc Inst, Div Hematol & Oncol, Detroit, MI 48202 USA. Bay State Med Ctr, Dept Obstet & Gynecol, Springfield, MA USA. Amer Canc Soc, Dept Epidemiol & Surveillance Res, Atlanta, GA 30329 USA. Emory Univ, Rollins Sch Publ Hlth, Dept Epidemiol, Atlanta, GA 30322 USA. Wayne State Univ, Karmanos Canc Inst, Div Epidemiol, Detroit, MI 48202 USA. Univ Penn, Dept Biostat & Epidemiol, Philadelphia, PA 19104 USA. Univ Penn, Ctr Clin Epidemiol & Biostat, Philadelphia, PA 19104 USA. Univ So Calif, Keck Sch Med, Dept Pathol, Los Angeles, CA USA. Univ So Calif, Keck Sch Med, Dept Prevent Med, Los Angeles, CA USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. RP Marchbanks, PA (reprint author), Ctr Dis Control & Prevent, Div Reprod Hlth MS K34, 1600 Clifton Rd, Atlanta, GA 30333 USA. FU NICHD NIH HHS [N01-HD-3-3175, N01-HD-2-3166, N01-HD-3-3168, N01-HD-3-3174, N01-HD-3-3176, Y01-HD-7022] NR 26 TC 108 Z9 109 U1 1 U2 5 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD MAY PY 2002 VL 12 IS 4 BP 213 EP 221 AR PII S1047-2797(01)00274-5 DI 10.1016/S1047-2797(01)00274-5 PG 9 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 543XY UT WOS:000175129300001 PM 11988408 ER PT J AU Shavers, VL Lynch, CF Burmeister, LF AF Shavers, VL Lynch, CF Burmeister, LF TI Racial differences in factors that influence the willingness to participate in medical research studies SO ANNALS OF EPIDEMIOLOGY LA English DT Article DE African American; blacks; minorities; medical research; participation; Tuskegee study; research barriers ID CANCER CLINICAL-TRIALS; AFRICAN-AMERICANS; MINORITY PARTICIPATION; RECRUITMENT; COMMUNITY; BLACK; AIDS; STRATEGIES; PREVENTION; EXPERIENCE AB PURPOSE: The relative absence of racial/ethnic minorities among medical research subjects is receiving considerable attention because of recent government mandates for their inclusion in all human subject re, search. We examined racial differences in the prevalence of sociocultural barriers as a possible explanation for the underrepresentation of African Americans in medical research studies. METHODS: During 1998-1999, a total of 198 residents of the Detroit Primary Metropolitan Statistical Area (PMSA) participated in a survey that examined impediments to participation in medical research studies. Chi square tests and logistic regression analyses were used to examine the association between race, issues related to trust of medical researchers, and the willingness to participate in medical research studies. RESULTS: Study results indicate that African Americans and whites differ in their willingness to participate in medical research. Racial differences in the willingness to participate in a medical research are primarily due to the lower level of trust of medical research among African Americans. African American respondents were also somewhat less willing to participate if they attribute high importance to the race of the doctor when seeking routine medical care, believed that minorities bear most of the risks of medical research, and if their knowledge of the Tuskegee Study resulted in less trust in medical researchers. CONCLUSION: These data reiterate the need for medical researchers to build trusting relationships with minority communities. Researchers can begin by acknowledging the previous medical abuse of minority research participants, discussing their specific plans to assure the protection of study participants, and explaining the need for the participation of racial/ethnic minorities including studies that specifically target or that are likely to result in disproportionate representation of racial/ethnic minorities among study participants. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Univ Iowa, Coll Publ Hlth, Dept Biostat, Iowa City, IA USA. Univ Iowa, Coll Publ Hlth, Dept Epidemiol, Iowa City, IA USA. Univ Iowa, Dept Epidemiol, Iowa City, IA USA. RP Shavers, VL (reprint author), NCI, Div Canc Control & Populat Sci, Appl Res Program, Hlth Serv & Econ Branch, Execut Plaza N,Room 4005,6130 Execut Blvd,MSC 734, Bethesda, MD 20892 USA. FU AHRQ HHS [R03 HS09597-01]; NCI NIH HHS [1F31 CA72125-01A1] NR 58 TC 264 Z9 264 U1 4 U2 24 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD MAY PY 2002 VL 12 IS 4 BP 248 EP 256 AR PII S1047-2797(01)00265-4 DI 10.1016/S1047-2797(01)00265-4 PG 9 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 543XY UT WOS:000175129300006 PM 11988413 ER PT J AU Zeuner, KE Bara-Jimenez, W Noguchi, PS Goldstein, SR Dambrosia, JM Hallett, M AF Zeuner, KE Bara-Jimenez, W Noguchi, PS Goldstein, SR Dambrosia, JM Hallett, M TI Sensory training for patients with focal hand dystonia SO ANNALS OF NEUROLOGY LA English DT Article ID TACTILE SPATIAL-RESOLUTION; PRIMARY SOMATOSENSORY CORTEX; REPETITIVE STRAIN INJURY; BLIND BRAILLE READERS; WRITERS CRAMP; CORTICAL REPRESENTATION; DISCRIMINATION; DISORDER; FINGERS; ACUITY AB Some patients with focal hand dystonia have impaired sensory perception. Abnormal sensory processing may lead to problems with fine motor control. For patients with focal hand dystonia who demonstrate sensory dysfunction, sensory training may reverse sensory impairment and dystonic symptoms. We studied the efficacy of learning to read braille as a method of sensory training for patients with focal hand dystonia. Sensory spatial discrimination was evaluated in 10 patients who had focal hand dystonia and 10 age- and gender-matched controls with a spatial acuity test (JVP domes were used in this test). Clinical dystonia evaluation included the Fahn dystonia scale and time needed to write a standard paragraph. Each individual was trained in braille reading at the grade 1 level for 8 weeks, between 30 and 60 minutes daily, and was monitored closely to ensure that reading was done regularly. Both controls and patients demonstrated improvement on the spatial acuity test. Patients showed a significant mean difference from baseline to 8 weeks on the Fahn dystonia scale. Sixty percent of the patients shortened the time they needed to write a standard paragraph. Improved sensory perception correlated positively with improvement on the Fahn dystonia scale. We conclude that training in braille reading improves deficits in spatial discrimination and decreases disability in patients with focal hand dystonia. C1 NINCDS, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. NINCDS, Biostat Branch, Human Motor Control Sect, Bethesda, MD 20892 USA. RP Hallett, M (reprint author), NINCDS, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. NR 36 TC 91 Z9 97 U1 0 U2 4 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD MAY PY 2002 VL 51 IS 5 BP 593 EP 598 DI 10.1002/ana.10174 PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 545PQ UT WOS:000175226700009 PM 12112105 ER PT J AU Smith, MJ Adams, LF Schmidt, PJ Rubinow, DR Wassermann, EM AF Smith, MJ Adams, LF Schmidt, PJ Rubinow, DR Wassermann, EM TI Effects of ovarian hormones on human cortical excitability SO ANNALS OF NEUROLOGY LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Neurological-Association CY OCT 01, 2001 CL CHICAGO, ILLINOIS SP Amer Neurolog Assoc ID TRANSCRANIAL MAGNETIC STIMULATION; DENDRITIC SPINE DENSITY; HUMAN MOTOR CORTEX; GABA(A) RECEPTOR-BINDING; FEMALE RAT-BRAIN; MENSTRUAL-CYCLE; INTRACORTICAL FACILITATION; NEUROACTIVE STEROIDS; POSTMENOPAUSAL WOMEN; ACTIVATION PATTERNS AB Ovarian steroids appear to alter neuronal function in women, but direct physiological evidence is lacking. In animals, estradiol enhances excitatory neurotransmission. Progesterone-derived neurosteroids increase GABAergic inhibition. The effect of weak transcranial magnetic stimulation of the motor cortex on the motor evoked potential (MEP) from transcranial magnetic stimulation given milliseconds later is changed by GABAergic and glutamatergic agents. Using this technique previously, we showed more inhibition in the luteal phase relative to the midfollicular menstrual phase, which is consistent with a progesterone effect. To detect the effects of estradiol, we have now divided the follicular phase. We tested 14 healthy women during the early follicular (low estradiol, low progesterone), late follicular (high estradiol, low progesterone), and luteal (high estradiol, high progesterone) phases, with interstimulus intervals from 2 to 10msec (10 trials at each interval and 40 unconditioned trials). We calculated the ratio of the conditioned MEP at each interval to the mean unconditioned MEP: the higher the ratio, the less inhibition and the more facilitation caused by the first stimulus. The combined ratios increased significantly from the early follicular phase to the late follicular phase and then decreased again in the luteal phase. These findings demonstrate an excitatory neuronal effect associated with estradiol and confirm our earlier finding of inhibition associated with progesterone. C1 NINCDS, Brain Stimulat Unit, NIH, Bethesda, MD 20892 USA. NIMH, Behav Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Wassermann, EM (reprint author), NINCDS, Brain Stimulat Unit, NIH, Bldg 10,Room 5N234, Bethesda, MD 20892 USA. NR 49 TC 140 Z9 142 U1 4 U2 10 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD MAY PY 2002 VL 51 IS 5 BP 599 EP 603 DI 10.1002/ana.10180 PG 5 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 545PQ UT WOS:000175226700010 PM 12112106 ER PT J AU Zhai, SP Senderowicz, AM Sausville, EA Figg, WD AF Zhai, SP Senderowicz, AM Sausville, EA Figg, WD TI Flavopiridol, a novel cyclin-dependent kinase inhibitor, in clinical development SO ANNALS OF PHARMACOTHERAPY LA English DT Article DE cyclin-dependent kinase; flavopiridol ID BREAST-CARCINOMA CELLS; ACTIVITY IN-VIVO; DRUG FLAVOPIRIDOL; CANCER; APOPTOSIS; L86-8275; INDUCTION; GROWTH; D1; OVEREXPRESSION AB OBJECTIVE: To review preclinical and clinical information on flavopiridol, an inhibitor of cyclin-dependent kinases (CDKs), tested as an antitumor agent. DATA SOURCES: Primary and review articles were identified by MEDLINE search (1 990-June 2001). Abstracts from recent meetings were also used as source materials. DATA EXTRACTION: Flavopiridol was reviewed with regard to its mechanisms, preclinical and clinical results, pharmacokinetics, and metabolism. DATA SYNTHESIS: Flavopiridol is an inhibitor of several CDKs and displays unique anticancer properties. In addition to direct CDK inhibition, flavopiridol also exhibited other features such as inducing apoptosis in many cancer cell lines, decreasing cyclin D1 concentration, and inhibiting angiogenesis. Preclinical xenograft models showed significant antitumor activity for flavopiridol. The regimen using 72-hour continuous infusion every 2 weeks has been most extensively applied in clinical trials, with a 1-hour infusion currently being explored to achieve higher peak concentrations. Several Phase I and II trials have been reported, with some evidence of antitumor activity noted. Further Phase I and II trials using flavopiridol as a single agent and in combination with standard chemotherapeutic regimens and various tumor types are ongoing. CONCLUSIONS: Flavopiridol is the first CDK inhibitor to enter clinical trials. Several Phase I and Phase II clinical trials with different regimens (less than or equal to72-h or 1-h infusion) have been completed. Initial clinical trials have been intriguing, but many questions remain: What is the best regimen (less than or equal to72-h infusion)? Does optimal future development of this drug depend on the combination with other chemotherapy? What is the best combination of flavopiridol with other chemotherapy? C1 NCI, Ctr Canc Res, Mol Pharmacol Sect, Bethesda, MD 20892 USA. NIDCR, Oral Pharyngeal Canc Branch, NIH, Bethesda, MD USA. NCI, Dev Therapeut Program, Bethesda, MD 20892 USA. RP Figg, WD (reprint author), NCI, Ctr Canc Res, Mol Pharmacol Sect, Bldg 10,Rm 5A01,MSC 1910,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 46 TC 87 Z9 93 U1 0 U2 3 PU HARVEY WHITNEY BOOKS CO PI CINCINNATI PA PO BOX 42696, CINCINNATI, OH 45242 USA SN 1060-0280 J9 ANN PHARMACOTHER JI Ann. Pharmacother. PD MAY PY 2002 VL 36 IS 5 BP 905 EP 911 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 548QM UT WOS:000175400200024 PM 11978170 ER PT J AU Kane, S Prentice, MA Mariano, JM Cuttitta, F Jakowlew, SB AF Kane, S Prentice, MA Mariano, JM Cuttitta, F Jakowlew, SB TI Differential induction of early response genes by adrenomedullin and transforming growth factor-beta 1 in human lung cancer cells SO ANTICANCER RESEARCH LA English DT Article DE adrenomedullin; TGF-beta; early response genes; AP-1 activity; lung cancer ID TISSUE-SPECIFIC EXPRESSION; SMOOTH-MUSCLE CELLS; STIMULATES C-FOS; FACTOR-BETA; PHORBOL ESTER; MESSENGER-RNA; ENDOTHELIAL-CELLS; MOUSE DEVELOPMENT; AP-1 ACTIVITY; MICE LACKING AB Adrenomedullin (AM) is a hypotensive polypeptide that has been shown to stimulate cyclic AMP and intracellular free Ca2+, agents that are known to induce expression of protooncogenes, in various cell types. Transforming growth factor-beta 1 (TGF-beta1) is a multifunctional polypeptide that regulates proliferation, differentiation and cell cycle progression in both normal and malignant epithelial cells. The diverse biological actions of AM and TGF-beta1 may be related to their capacities to initiate different genomic programs in target cells via the induction of expression of multiple genes including early response genes and proto-oncogenes. AM, TGFbeta1 and phorbol-12-myristate-13-acetate (PMA) exert both positive and negative effects on mitogenesis. The effects of AM, TGF-beta1 and PMA were examined in human non-small cell lung cancer (NSCLC) cells. AM caused an increase in its mRNA transcript that peaked by 6 hours and persisted to 24 hours. While expression of TGF-beta1 mRNA was not affected by AM in these cells, the mRNAs for TGF-beta1 and TGF-beta3 decreased by 3 hours In contrast, TGF-beta1 had no effect on expression of AM mRNA. Interestingly, PMA caused an increase in AM and TGF-beta1 mRNAs in NSCLC cells. While both TGF-beta1 and PMA caused a transient increase in expression of the mRNAs for early response genes including c-fos, c-jun and egr-1 that peaked by 1 hour following treatment, the increase in expression of these mRNAs following treatment with AM peaked only after 3-6 hours. Western blotting analysis showed increases in the levels of c-jun protein following treatment with AM, TGF-beta1 and PMA. The increase in c-jun protein from treatment with AM occurred 10 hours after that from TGF-beta1 and PMA. Activator protein 1 (AP-1) DNA binding activity was. also demonstrated to increase following treatment with AM, TGF-beta1 and PMA, the increase in AP-1 DNA binding activiy following AM treatment occurring 10 hours later than that from TGF-beta1 and PMA treatment. These data show that AM can regulate expression of its mRNA transcript in NSCLC cells. Our study suggests that NSCLC cells are important targets of AM and TGF-beta1 and that AM and TGF-beta1 may regulate activities in these malignant lung cells through differential induction of various early response genes. C1 NCI, Cell & Canc Biol Branch, Rockville, MD 20850 USA. RP Jakowlew, SB (reprint author), NCI, Cell & Canc Biol Branch, 9610 Med Ctr Dr,Suite 300, Rockville, MD 20850 USA. NR 77 TC 16 Z9 17 U1 0 U2 1 PU INT INST ANTICANCER RESEARCH PI ATHENS PA EDITORIAL OFFICE 1ST KM KAPANDRITIOU-KALAMOU RD KAPANDRITI, PO BOX 22, ATHENS 19014, GREECE SN 0250-7005 J9 ANTICANCER RES JI Anticancer Res. PD MAY-JUN PY 2002 VL 22 IS 3 BP 1433 EP 1444 PG 12 WC Oncology SC Oncology GA 580RA UT WOS:000177246500011 PM 12168820 ER PT J AU Ju, JY Polhamus, C Marr, KA Holland, SM Bennett, JE AF Ju, JY Polhamus, C Marr, KA Holland, SM Bennett, JE TI Efficacies of fluconazole, caspofungin, and amphotericin B in Candida glabrata-infected p47(phox-/-) knockout mice SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID CHRONIC GRANULOMATOUS-DISEASE; TORULOPSIS-GLABRATA; HOST-DEFENSE; IN-VITRO; FUNGEMIA; SUSCEPTIBILITY; L-743,872; KRUSEI; MODEL AB Candida glabrata is the second leading cause of adult candidemia, resulting in high mortality. Amphotericin B is considered the treatment of choice, while the efficacy of fluconazole is controversial and caspofungin efficacy is unknown. To ascertain drug efficacy in vivo, the utility of a murine model of C. glabrata infection was investigated. C. glabrata was found to cause progressive, lethal infection when injected intravenously into C57BL/6 mice with reduced oxidative microbicidal capacity due to knockout of the p47(phox) gene. Spleen and kidney organ CFU counts were determined in groups of mice 2 days after the mice completed 6 days of daily intraperitoneal drug treatment, which began on the day of infection. Daily injections of fluconazole at 80 mg/kg did not reduce spleen or kidney CFU counts after infection with C. glabrata strains having in vitro fluconazole MICs of 2, 32, or 256 mug/ml compared to saline-treated controls. However, this fluconazole regimen reduced spleen CFU counts in mice infected with Candida albicans, an infection that is known to be responsive to fluconazole. Caspofungin at 5 mg/kg and amphotericin B at 5 mg/kg were both effective in reducing fungal burden in spleens and kidneys of C. glabrata-infected mice. Ten mice treated for 6 days with caspofungin at I mg/kg survived for 15 days, though all 10 saline-injected mice died or were so ill that they had to be sacrificed by 96 h postinfection. This murine model provided evidence of the efficacy of amphotericin B and caspofungin but not of fluconazole against C. glabrata infection. C1 NIAID, Clin Mycol Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. Univ Washington, Sch Med, Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA. RP Bennett, JE (reprint author), NIH, Ctr Clin, Rm 11C304, Bethesda, MD 20892 USA. NR 22 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 0066-4804 J9 ANTIMICROB AGENTS CH JI Antimicrob. Agents Chemother. PD MAY PY 2002 VL 46 IS 5 BP 1240 EP 1245 DI 10.1128/AAC.46.5.1240-1245.2002 PG 6 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 543KH UT WOS:000175100800011 PM 11959551 ER PT J AU Powell, MB Roberts, KP AF Powell, MB Roberts, KP TI The effect of repeated experience on children's suggestibility across two question types SO APPLIED COGNITIVE PSYCHOLOGY LA English DT Article ID MEMORY TRACE STRENGTH; EVENT MEMORY; DEVELOPMENTAL-CHANGES AB There is a discrepancy in the literature regarding the effect of repeated experience on children's suggestibility. Some researchers have concluded that repeated experience increases children's suggestibility for variable details whereas others have reported no detrimental effect. This study demonstrated that the type of question used to test memory (cued-recall versus yes/no questions) could account for the different reported conclusions. Children aged 5-6 years took part in an event either once or four times. Three or 21 days later, they were given a suggestive interview about the single/final occurrence of the event during which half of the event details were inaccurately described. When later asked yes/no questions, the children with repeated experience agreed with more of the suggestions than did those in the single-experience condition, especially at the longer delay. In relation to cued-recall questions, however, experience did not mediate the number of times that false suggestions from the biasing interview were reported. This latter finding was revealed irrespective of the retention interval. Copyright (C) 2002 John Wiley Sons, Ltd. C1 Deakin Univ, Sch Psychol, Burwood 3125, Australia. NICHHD, Bethesda, MD 20892 USA. RP Powell, MB (reprint author), Deakin Univ, Sch Psychol, 221 Burwood Hwy, Burwood 3125, Australia. NR 27 TC 29 Z9 30 U1 0 U2 5 PU JOHN WILEY & SONS LTD PI CHICHESTER PA THE ATRIUM, SOUTHERN GATE, CHICHESTER PO19 8SQ, W SUSSEX, ENGLAND SN 0888-4080 J9 APPL COGNITIVE PSYCH JI Appl. Cogn. Psychol. PD MAY PY 2002 VL 16 IS 4 BP 367 EP 386 DI 10.1002/acp.801 PG 20 WC Psychology, Experimental SC Psychology GA 696EL UT WOS:000183873500001 ER PT J AU Hershkowitz, I Orbach, Y Lamb, ME Sternberg, KJ Horowitz, D AF Hershkowitz, I Orbach, Y Lamb, ME Sternberg, KJ Horowitz, D TI A comparison of mental and physical context reinstatement in forensic interviews with alleged victims of sexual abuse SO APPLIED COGNITIVE PSYCHOLOGY LA English DT Article ID COGNITIVE INTERVIEW; CHILDRENS MEMORY; EYEWITNESS MEMORY; INVESTIGATIVE INTERVIEWS; WITNESSES; EVENT; ENHANCEMENT; RESPONSES; STRESSFUL; TESTIMONY AB Forensic interviews with 142 alleged victims of sexual abuse, ranging from 4.0 to 13.5 years of age, were conducted under three different interviewing conditions: at the scene of the incidents (Physical Context Reinstatement; PCR group), in the investigator's office with mental context reinstating instructions (MCR group), and in the office without contextual cueing (Control group). Children in the PCR, MCR, and Control groups did not differ significantly with respect to the total number of informative details reported. Children in the MCR condition provided more detailed responses to the main invitation and in their first narrative than did children in the PCR condition, however. They also provided proportionally more detailed responses to open-ended invitations and fewer details in response to directive prompts than did children in the two other conditions. The MCR procedures were thus associated with greater improvements in the quality of information retrieval than were the PCR procedures. In all interviewing conditions, children aged 7 to 13 years provided significantly more details than 4- to-6-year-old children did. The youngest children provided fewer details in response to invitations and directive utterances, and proportionally more details in response to option-posing and suggestive utterances. No significant interactions between age and interviewing condition were apparent. Published in 2002 by John Wiley Sons, Ltd. C1 NICHHD, Sect Social & Emot Dev, Bethesda, MD 20892 USA. Minist Labor & Social Affairs, Jerusalem, Israel. Univ Haifa, IL-31999 Haifa, Israel. RP Lamb, ME (reprint author), NICHHD, Sect Social & Emot Dev, Rockledge 1 Ctr,Suite 8048,6705 Rockledge Dr, Bethesda, MD 20892 USA. NR 58 TC 20 Z9 20 U1 4 U2 13 PU JOHN WILEY & SONS LTD PI CHICHESTER PA THE ATRIUM, SOUTHERN GATE, CHICHESTER PO19 8SQ, W SUSSEX, ENGLAND SN 0888-4080 J9 APPL COGNITIVE PSYCH JI Appl. Cogn. Psychol. PD MAY PY 2002 VL 16 IS 4 BP 429 EP 441 DI 10.1002/acp.804 PG 13 WC Psychology, Experimental SC Psychology GA 696EL UT WOS:000183873500004 ER PT J AU Carter, MC Metcalfe, DD AF Carter, MC Metcalfe, DD TI Paediatric mastocytosis SO ARCHIVES OF DISEASE IN CHILDHOOD LA English DT Article ID MAST-CELL DISEASE; GASTROINTESTINAL STROMAL TUMORS; ACUTE LYMPHOBLASTIC-LEUKEMIA; PROTOONCOGENE C-KIT; SYSTEMIC MASTOCYTOSIS; URTICARIA-PIGMENTOSA; BONE-MARROW; DISODIUM-CROMOGLYCATE; PEDIATRIC MASTOCYTOSIS; BULLOUS MASTOCYTOSIS C1 NIAID, NIH, LAD, Bethesda, MD 20892 USA. RP Carter, MC (reprint author), NIAID, NIH, LAD, Bldg 10,Room 11-C206,10 Ctr Dr MSC 1881, Bethesda, MD 20892 USA. NR 73 TC 37 Z9 40 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0003-9888 J9 ARCH DIS CHILD JI Arch. Dis. Child. PD MAY PY 2002 VL 86 IS 5 BP 315 EP 319 DI 10.1136/adc.86.5.315 PG 5 WC Pediatrics SC Pediatrics GA 548MG UT WOS:000175391300001 PM 11970917 ER PT J AU Spencer, HB Hussein, WR Tchounwou, PB AF Spencer, HB Hussein, WR Tchounwou, PB TI Effects of tetrachloroethylene on the viability and development of embryos of the Japanese medaka, Oryzias latipes SO ARCHIVES OF ENVIRONMENTAL CONTAMINATION AND TOXICOLOGY LA English DT Article ID TOXICITY; TERATOGENICITY; SYSTEM; FISH; EGGS AB We evaluated the acute toxicity of Tetrachloroethylene (C2Cl4), and investigated its sub-chronic effects on the embryonic development of Japanese medaka (Oryzias latipes). One-day-old eggs/embryos of this fish species were exposed, under static renewal conditions, to serial concentrations (0, 20, 40, 60, and 80 mg/L) of C2Cl4 for 96 h (acute) and 10 days (sub-chronic) time periods. The toxic endpoints evaluated included: egg/embryo viability, hatchability, and morphological/developmental abnormalities. The acute toxicity test resulted in a 96 h-LC50 of 27.0 (19.5-32.9) mg/L for egg viability. Exposure of eggs to sub-chronic concentrations (0, 1.5, 3, 6, 12, and 25 mg/L) of C2Cl4 significantly reduced hatchability and larval survival, in a concentration dependent manner. At the highest tested concentration (25 mg/L) of the sub-lethal exposure, larval survival was greatly reduced to within three days post-hatch. The lowest tested concentration (1.5 mg/L) produced a significant number of developmental effects to the Japanese medaka, including abnormal development of the circulatory system, yolk-sac edema, pericardial edema, scoliosis, hemorrhaging, blood pooling, and defects in heart morphology. The severity of these abnormalities was concentration-dependent, It can be concluded from these results that tetrachloroethylene is teratogenic to the Japanese medaka. C1 Jackson State Univ, Environm Toxicol Res Lab, Ctr Environm Hlth, Sch Sci & Technol,NIH, Jackson, MS 39217 USA. Jackson State Univ, Sch Sci & Technol, Dept Chem, Jackson, MS 39217 USA. RP Tchounwou, PB (reprint author), Jackson State Univ, Environm Toxicol Res Lab, Ctr Environm Hlth, Sch Sci & Technol,NIH, 1400 Lynch St,POB 18540, Jackson, MS 39217 USA. NR 24 TC 7 Z9 8 U1 1 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0090-4341 J9 ARCH ENVIRON CON TOX JI Arch. Environ. Contam. Toxicol. PD MAY PY 2002 VL 42 IS 4 BP 463 EP 469 DI 10.1007/s00244-001-0050-1 PG 7 WC Environmental Sciences; Toxicology SC Environmental Sciences & Ecology; Toxicology GA 544TF UT WOS:000175176200011 PM 11994788 ER PT J AU Moore, LE Lu, M Smith, AH AF Moore, LE Lu, M Smith, AH TI Childhood cancer incidence and arsenic exposure in drinking water in Nevada SO ARCHIVES OF ENVIRONMENTAL HEALTH LA English DT Article DE ALL; arsenic; childhood cancer; cluster; drinking water ID ACUTE LYMPHOBLASTIC-LEUKEMIA; EXFOLIATED BLADDER CELLS; SODIUM ARSENITE; MORTALITY; CHILDREN; MICRONUCLEI; INDIVIDUALS; ARGENTINA; RADIATION; DISEASE AB Inorganic arsenic exposure through drinking water causes cancer in adults; however, the carcinogenic potential in children remains unknown. A recent leukemia cluster in Churchill County, Nevada, where arsenic levels in water supplies are relatively high, has prompted concern. The authors investigated the incidence of childhood cancer between 1979 and 1999 in all 17 Nevada counties, grouped by low (i.e., < 10 mug/l), medium (10-25 mug/l), and high (35-90 mug/l) population-weighted arsenic levels in public drinking water supplies. The standardized incidence ratios (SIRs) for all childhood cancers combined were 1.00 (95% confidence interval [CI] = 0.94,11.06), 0.72 (95% CI = 0.43, 1.12), and 1.25 (95% CI = 0.91, 1.69) for low-, medium-, and high-exposure counties, respectively. There was no relationship between arsenic levels in water and childhood leukemia (SIRs = 1.02, 0.61, and 0.86, respectively [95% CIs = 0.90, 1.15; 0.12, 1.79; and 0.37, 1.70, respectively]). For all childhood cancers, excluding leukemias, the SIRs were 0.99 (95% CI = 0.92, 1.07), 0.82 (95% CI = 0.42, 1.22), and 1.37 (0.92, 1.83), respectively. The excess in 5- to 9-yr-old children and 10- to 14-yr-old children was in bone cancers, and the excess in 15- to 19-yr-old young adults was primarily in lymphomas. The findings in this study are reassuring in that leukemia risks were not increased at the concentrations of arsenic in water found in this study. Nonetheless, the results raise the possibility that there are increased risks for non-leukemic childhood cancers that require confirmation in other studies, particularly those in which higher exposures are addressed. C1 NCI, Occupat Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ Calif Berkeley, Sch Publ Hlth, Berkeley, CA 94720 USA. RP Moore, LE (reprint author), NCI, Occupat Epidemiol Branch, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS 8118,MSC 7240, Bethesda, MD 20892 USA. RI Smith, Allan/F-9249-2011 NR 30 TC 33 Z9 33 U1 0 U2 4 PU HELDREF PUBLICATIONS PI WASHINGTON PA 1319 EIGHTEENTH ST NW, WASHINGTON, DC 20036-1802 USA SN 0003-9896 J9 ARCH ENVIRON HEALTH JI Arch. Environ. Health PD MAY-JUN PY 2002 VL 57 IS 3 BP 201 EP 206 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health GA 628FR UT WOS:000179982400004 PM 12507173 ER PT J AU Katz, ML Robison, WG AF Katz, ML Robison, WG TI What is lipofuscin? Defining characteristics and differentiation from other autofluoreseent lysosomal storage bodies SO ARCHIVES OF GERONTOLOGY AND GERIATRICS LA English DT Article; Proceedings Paper CT 6th International Symposium on Lipofuscin and Ceroid Pigments CY MAY 19-22, 2001 CL UNIV HAWAII, HONOLULU, HAWAII HO UNIV HAWAII DE lipofuscin; macular degeneration; retina; retinal pigment epithelium; ceroid ID RETINAL-PIGMENT EPITHELIUM; VITAMIN-E-DEFICIENCY; FLUORESCENT PIGMENT; LIPID-PEROXIDATION; VISUAL CYCLE; A COMPOUNDS; HUMAN RPE; RAT; ACCUMULATION; PHOTORECEPTOR AB Lipofuscins, also known as age-pigments, have three defining characteristics: (1) they consist of intracellular secondary lysosomes; (2) they have a yellow autofluorescent emission when excited by near ultraviolet or blue light; and (3) they accumulate during normal senescence. Lysosomal storage bodies with similar fluorescence properties accumulate in various cell types as a result of specific pathological conditions or experimental manipulations. As a class, the latter are often referred to as ceroid pigments. In general, the mechanisms involved in the formation of ceroid pigments cannot be assumed to be closely similar to those involved in lipofuscin formation. In fact, the mechanisms of formation almost certainly differ, not only between lipofuscins and ceroids, but also among different lipofuscins and different ceroids. Presently, the most detailed knowledge about the mechanisms involved in lipofuscin formation come from studies on the retinal pigment epithelium (RPE) of the eye. These studies indicate that at least the autofluorescent constituents of RPE lipofuscin are generated from derivatives of vitamin A that occur in the retina. Oxidative stress to the retina appears to promote the formation of these RPE fluorophores. Whether similar mechanisms are involved in the formation of the lipofuscins that occur in other tissues remains to be determined, The mechanisms involved in RPE lipofuscin fluorophore formation are closely related to metabolic pathways that are specific to the retina. Thus, it appears likely that the mechanisms by which lipofuscins form in other tissues differ fundamentally from those that underlie RPE lipofuscin formation. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Missouri, Sch Med, Mason Eye Inst, Columbia, MO 65212 USA. NEI, NIH, Bethesda, MD 20892 USA. RP Robison, WG (reprint author), Univ Missouri, Sch Med, Mason Eye Inst, 1 Hosp Dr, Columbia, MO 65212 USA. NR 54 TC 43 Z9 45 U1 0 U2 6 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0167-4943 J9 ARCH GERONTOL GERIAT JI Arch. Gerontol. Geriatr. PD MAY-JUN PY 2002 VL 34 IS 3 BP 169 EP 184 AR PII S0167-4943(02)00005-5 DI 10.1016/S0167-4943(02)00005-5 PG 16 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 539PY UT WOS:000174880700002 PM 14764321 ER PT J AU Nachman, SA Lindsey, JC Pelton, S Mofenson, L McIntosh, K Wiznia, A Stanley, K Yogev, R AF Nachman, SA Lindsey, JC Pelton, S Mofenson, L McIntosh, K Wiznia, A Stanley, K Yogev, R TI Growth in human immunodeficiency virus-infected children receiving ritonavir-containing antiretroviral therapy SO ARCHIVES OF PEDIATRICS & ADOLESCENT MEDICINE LA English DT Article; Proceedings Paper CT 39th Interdisciplinary Conference on Antimicrobial Agents and Chemotherapy (ICAAC) CY SEP 26-29, 1999 CL SAN FRANCISCO, CALIFORNIA ID PROTEASE INHIBITORS; BODY-COMPOSITION; SOMATIC GROWTH; HIV-INFECTION; WEIGHT-GAIN; TYPE-1 RNA; DISEASE; BORN; STANDARD; MOTHERS AB Background: Human immunodeficiency virus (HIV)infected children often suffer from impaired growth. Highly active antiretroviral therapy (HAART) successfully reduces HIV 1 (HIV-1) RNA to 400 copies/mL or less in many children. Objectives: To determine if age- and sex-adjusted growth z scores correlate with HIV-1 RNA level and if control of viral load for 48 and 96 weeks results in improved growth in children receiving highly active antiretroviral therapy. Design: Secondary analysis of the cohort of children receiving ritonavir nested in a randomized, open-label, clinical trial. Subjects and Methods: The Pediatric AIDS Clinical Trials Group Protocol 338 enrolled clinically stable, antiretroviral therapy-experienced, HIV-infected subjects aged 2 through 17 years, Using data from subjects randomized to ritonavir-containing regimens (n = 197), the association of growth z scores and HIV-1 RNA levels were examined. Main Outcome Measures: Age- and sex-adjusted weight and height z scores. Results: Enrollment weights were comparable with age- and sex-adjusted norms, but subjects receiving ritonavir-containing antiretroviral therapy were significantly shorter (mean z score, -0.57 [29th percentile] 95% confidence interval, -0.73 to -0.40). Higher HIV-1 RNA levels correlated with lower growth z scores (P<.01). Subjects achieving and maintaining HIV-1 RNA of 400 copies/mL or less through 48 and 96 weeks experienced worse growth than subjects with a less controlled viral load. Conclusions: In this pediatric cohort, a significant decline in height and weight z scores was found despite control of viral replication. Further studies of growth are necessary to assess if nutritional and hormonal adjuvants to highly active antiretroviral therapy should be considered to improve growth in HIV-infected children. C1 SUNY Stony Brook, Hlth Sci Ctr, Dept Pediat, Stony Brook, NY 11794 USA. Harvard Sch Publ Hlth, Ctr Biostat AIDS Res, Boston, MA USA. Boston Med Ctr, Dept Pediat, Boston, MA USA. NICHHD, Pediat Adolescent & Maternal AIDS Branch, NIH, Rockville, MD USA. Childrens Hosp Boston, Dept Pediat, Boston, MA USA. Albert Einstein Med Ctr, Jacobi Hosp, Dept Pediat, Bronx, NY USA. Northwestern Univ, Childrens Mem Hosp, Dept Pediat, Chicago, IL 60614 USA. RP Nachman, SA (reprint author), SUNY Stony Brook, Hlth Sci Ctr, Dept Pediat, Stony Brook, NY 11794 USA. OI Mofenson, Lynne/0000-0002-2818-9808 FU NIAID NIH HHS [AI-41110] NR 34 TC 24 Z9 24 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 1072-4710 J9 ARCH PEDIAT ADOL MED JI Arch. Pediatr. Adolesc. Med. PD MAY PY 2002 VL 156 IS 5 BP 497 EP 503 PG 7 WC Pediatrics SC Pediatrics GA 548RC UT WOS:000175402100017 PM 11980557 ER PT J AU Jaffer, FA O'Donnell, CJ Larson, MG Chan, SK Kissinger, KV Kupka, MJ Salton, C Botnar, RM Levy, D Manning, WJ AF Jaffer, FA O'Donnell, CJ Larson, MG Chan, SK Kissinger, KV Kupka, MJ Salton, C Botnar, RM Levy, D Manning, WJ TI Age and sex distribution of subclinical aortic atherosclerosis - A magnetic resonance imaging examination of the Framingham Heart Study SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Article DE magnetic resonance imaging; aortas; atherosclerosis; imaging; subclinical disease ID BEAM COMPUTED-TOMOGRAPHY; CORONARY-ARTERY DISEASE; INTIMA-MEDIA THICKNESS; MYOCARDIAL-INFARCTION; OLDER ADULTS; RISK-FACTORS; CARDIOVASCULAR-DISEASE; HIGH-RESOLUTION; THORACIC AORTA; BLOOD-PRESSURE AB Autopsy data demonstrate a correlation between subclinical aortic atherosclerosis and cardiovascular disease. Therefore, noninvasive cardiovascular magnetic resonance (CMR) of subclinical atherosclerosis may provide a novel measure of cardiovascular risk, but it has not been applied to an asymptomatic population-based cohort to establish age- and sex-specific normative data. Participants in the Framingham Heart Study offspring cohort who were free of clinically apparent coronary disease were randomly sampled from strata of sex, quartiles of age, and quintiles of Framingham Coronary Risk Score. Subjects (n=318, aged 60 9 years, range 36 to 78 years, 51% women) underwent ECG-gated T2-weighted black-blood thoracoabdominal aortic CMR scanning. CMR evidence of aortic atherosclerosis was noted in 38% of the women and 41% of the men. Plaque prevalence and all measures of plaque burden increased with age group and were greater in the abdomen than in the thorax for both sexes and across all age groups. In addition, the Framingham Coronary Risk Score was significantly correlated with all plaque prevalence and burden measures for women but only for men after age adjustment. These noninvasive CMR data extend the prior autopsy-based prevalence estimates of subclinical atherosclerosis and may help to lay the foundation for future studies of risk stratification and treatment of affected individuals. C1 Harvard Univ, Beth Israel Deaconess Med Ctr, Sch Med, Dept Med,Cardiovasc Div,Harvard Thorndike Lab, Boston, MA 02215 USA. Harvard Univ, Beth Israel Deaconess Med Ctr, Sch Med, Dept Radiol, Boston, MA 02215 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Internal Med,Div Cardiol, Boston, MA USA. RP Manning, WJ (reprint author), Harvard Univ, Beth Israel Deaconess Med Ctr, Sch Med, Dept Med,Cardiovasc Div,Harvard Thorndike Lab, 330 Brookline Ave, Boston, MA 02215 USA. RI Botnar, Rene/E-6875-2012; OI Botnar, Rene/0000-0003-2811-2509; Jaffer, Farouc/0000-0001-7980-384X; Larson, Martin/0000-0002-9631-1254 FU NHLBI NIH HHS [N01-HC-38038] NR 27 TC 124 Z9 127 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD MAY PY 2002 VL 22 IS 5 BP 849 EP 854 DI 10.1161/01.ATV.0000012662.29622.00 PG 6 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 550PP UT WOS:000175511700025 PM 12006401 ER PT J AU Jacobs, DR Gross, MD Tracy, R Lewis, CE Sidney, S Liu, K Loria, C Steffes, MW AF Jacobs, DR Gross, MD Tracy, R Lewis, CE Sidney, S Liu, K Loria, C Steffes, MW TI Oxidative damage and inflammation are separate pathways for early coronary calcification SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Meeting Abstract C1 Univ Minnesota, Minneapolis, MN 55455 USA. Univ Vermont, Colchester, VT USA. Univ Alabama, Birmingham, AL USA. Kaiser Permanente, Div Res, Oakland, CA USA. Northwestern Univ, Sch Med, Chicago, IL 60611 USA. NHLBI, 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 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD MAY PY 2002 VL 22 IS 5 MA P136 BP A24 EP A24 PG 1 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 550PP UT WOS:000175511700160 ER PT J AU Liang, JS Fang, SY Weissman, AM Ginsberg, HN AF Liang, JS Fang, SY Weissman, AM Ginsberg, HN TI Overexpression of the tumor autocrine motility factor receptor-gp78, ubiquitin protein ligase (E3), results in increased ubiquitination and decreased secretion of apolipoprotein B in Hep G2 cells SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Meeting Abstract C1 Columbia Univ, Coll Phys & Surg, New York, NY 10027 USA. NCI, Ctr Canc Res, Regulat Prot Funct Lab, 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 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD MAY PY 2002 VL 22 IS 5 MA P442 BP A77 EP A77 PG 1 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 550PP UT WOS:000175511700450 ER PT J AU Wang, MY Takagi, G Asai, K Vatner, DE Natividad, FF Lakatta, EG AF Wang, MY Takagi, G Asai, K Vatner, DE Natividad, FF Lakatta, EG TI Altered matrix metalloprotease-2 regulation and tissue angiotensin and age-associated aortic remodeling in non-human primates SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Meeting Abstract C1 NIA, Baltimore, MD 21224 USA. Hackensack Univ, Med Ctr, Hackensack, NJ 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 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD MAY PY 2002 VL 22 IS 5 MA P429 BP A75 EP A75 PG 1 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 550PP UT WOS:000175511700438 ER PT J AU Zhao, B Huang, W Zhang, WY Kruth, HS AF Zhao, B Huang, W Zhang, WY Kruth, HS TI Interaction of aggregated LDL with cultured human coronary artery endothelial cells SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Meeting Abstract C1 NHLBI, Sect Expt Atherosclerosis, NIH, 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 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD MAY PY 2002 VL 22 IS 5 MA P49 BP A9 EP A9 PG 1 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 550PP UT WOS:000175511700078 ER PT J AU Huizinga, TWJ Machold, KP Breedveld, FC Lipsky, PE Smolen, JS AF Huizinga, TWJ Machold, KP Breedveld, FC Lipsky, PE Smolen, JS TI Criteria for early rheumatoid arthritis - From Bayes' law revisited to new thoughts on pathogenesis SO ARTHRITIS AND RHEUMATISM LA English DT Article ID COMBINATION THERAPY; POLYARTHRITIS C1 Leiden Univ, Med Ctr, Dept Rheumatol, NL-2300 RC Leiden, Netherlands. Univ Vienna, A-1010 Vienna, Austria. Natl Inst Hlth, Bethesda, MD USA. RP Huizinga, TWJ (reprint author), Leiden Univ, Med Ctr, Dept Rheumatol, C4 R POB 9600, NL-2300 RC Leiden, Netherlands. NR 14 TC 40 Z9 41 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 0004-3591 J9 ARTHRITIS RHEUM JI Arthritis Rheum. PD MAY PY 2002 VL 46 IS 5 BP 1155 EP 1159 DI 10.1002/art.10195 PG 5 WC Rheumatology SC Rheumatology GA 552XU UT WOS:000175646000004 PM 12115216 ER PT J AU Girschick, HJ Grammer, AC Nanki, T Vazquez, E Lipsky, PE AF Girschick, HJ Grammer, AC Nanki, T Vazquez, E Lipsky, PE TI Expression of recombination activating genes 1 and 2 in peripheral B cells of patients with systemic lupus erythematosus SO ARTHRITIS AND RHEUMATISM LA English DT Article ID MURINE RAG-2 PROMOTER; ANTI-DNA ANTIBODIES; V(D)J RECOMBINATION; T-CELL; GERMINAL-CENTERS; HUMAN TONSIL; CD40 LIGAND; RECEPTOR; LYMPHOCYTES; RESPONSES AB Objective. To analyze immunoregulatory abnormalities in patients with systemic lupus erythematosus (SLE) by assessing the expression of messenger RNA (mRNA) for types 1 and 2 recombination activating genes (RAG) in the peripheral blood of patients with active SLE. Methods. We examined B cell populations and also individual B cells from patients with SLE for the expression of RAG mRNA. Results. Analysis of bulk mRNA indicated that RAG1 and RAG2 mRNA were found routinely in peripheral B cells of patients with active SLE, but not in healthy subjects. When assessed on a single-cell basis, there was a 3-fold increase in the frequency of RAG1 and RAG2-expressing B cells in SLE patients compared with healthy subjects. Notably, B cells expressing both RAG1 and RAG2 mRNA expressed only IgD mRNA, but not IgG mRNA. Fifty percent of RAG-expressing B cells also expressed VpreB mRNA, whereas all expressed CD154 mRNA. Phenotypic analysis indicated that RAG-expressing B cells were activated, mature B cells. Conclusion. These results indicate that RAG expression is up-regulated in peripheral IgD+ and VpreB+ B cells of patients with active SLE. These cells may contribute to the immunoregulatory abnormalities in patients with SLE. C1 Univ Texas, SW Med Ctr, Dallas, TX 75230 USA. NIAMSD, NIH, Bethesda, MD 20892 USA. RP Lipsky, PE (reprint author), NIAMS, NIH, 9000 Rockville Pike,Bldg 10,Room 9N228, Bethesda, MD 20892 USA. FU NIAID NIH HHS [AI-31229] NR 61 TC 25 Z9 28 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 0004-3591 J9 ARTHRITIS RHEUM JI Arthritis Rheum. PD MAY PY 2002 VL 46 IS 5 BP 1255 EP 1263 DI 10.1002/art.10264 PG 9 WC Rheumatology SC Rheumatology GA 552XU UT WOS:000175646000019 PM 12115231 ER PT J AU Hoffman, GS Cid, MC Hellman, DB Guillevin, L Stone, JH Schousboe, J Cohen, P Calabrese, LH Dickler, H Merkel, PA Fortin, P Flynn, JA Locker, GA Easley, KA Schned, E Hunder, GG Sneller, MC Tuggle, C Swanson, H Hernandez-Rodriguez, J Lopez-Soto, A Bork, D Hoffman, DB Kalunian, K Klashman, D Wilke, WS Scheetz, RJ Mandell, BF Fessler, BJ Kosmorsky, G Prayson, R Luqmani, RA Nuki, G McRorie, E Sherrer, Y Baca, S Walsh, B Ferland, D Soubrier, M Choi, HK Gross, W Segal, AM Ludivico, C Puechal, X AF Hoffman, GS Cid, MC Hellman, DB Guillevin, L Stone, JH Schousboe, J Cohen, P Calabrese, LH Dickler, H Merkel, PA Fortin, P Flynn, JA Locker, GA Easley, KA Schned, E Hunder, GG Sneller, MC Tuggle, C Swanson, H Hernandez-Rodriguez, J Lopez-Soto, A Bork, D Hoffman, DB Kalunian, K Klashman, D Wilke, WS Scheetz, RJ Mandell, BF Fessler, BJ Kosmorsky, G Prayson, R Luqmani, RA Nuki, G McRorie, E Sherrer, Y Baca, S Walsh, B Ferland, D Soubrier, M Choi, HK Gross, W Segal, AM Ludivico, C Puechal, X CA INSSYS TI A multicenter, randomized, double-blind, placebo-controlled trial of adjuvant methotrexate treatment for giant cell arteritis SO ARTHRITIS AND RHEUMATISM LA English DT Article ID LOW-DOSE METHOTREXATE; PERMANENT VISUAL-LOSS; TEMPORAL ARTERITIS; POLYMYALGIA-RHEUMATICA; WEGENERS-GRANULOMATOSIS; CORTICOSTEROID REGIMENS; TAKAYASU ARTERITIS; DISEASE; BIOPSY; COMPLICATIONS AB Objective. To evaluate treatment with methotrexate (MTX) in patients with newly diagnosed giant cell arteritis (GCA) to determine if MTX reduces GCA relapses and cumulative corticosteroid (CS) requirements and diminishes disease- and treatment-related morbidity. Methods. This was a multicenter, randomized, double-blind study. Over 4 years, 16 centers from the International Network for the Study of Systemic Vasculitides enrolled patients with unequivocal GCA. The initial treatment was 1 mg/kg/day (less than or equal to60 mg every day) prednisone, plus either 0.15 mg/kg/week MTX (increased to 0.25 mg/kg/week, for a maximum weekly dosage of 15 mg) or placebo. Two physicians, both blinded to treatment allocation, evaluated each patient at every trial visit. One physician was responsible for providing global medical care. The other assessed GCA status according to a standard protocol. Treatment failure was defined as 2 distinct relapses or persistence of disease activity after the first relapse, in spite of increased CS therapy. Results. Ninety-eight patients were enrolled. No significant differences between treatment groups were noted with regard to age, frequency of positive findings on temporal artery biopsy (placebo 87%, MTX 79%), or comorbidities at the time of enrollment. The median dosage of MTX was 15 mg/week. The incidence of treatment failure was comparable between groups after 12 months: 57.5% in the MTX group failed treatment (95% confidence interval [95% CI] 41.6-73.4%) compared with 77.3% in the placebo group (95% CI 61.9-92.8%) (P = 0.26). In a Cox regression analysis, MTX was not associated with a reduced risk of treatment failure (relative risk 0.72; 95% CI 0.41-1.28). There were no significant differences between groups with regard to abnormal elevations of the erythrocyte sedimentation rate following initial remissions, serious morbidity due to GCA, cumulative CS dose, or treatment toxicity. In the MTX group, there were fewer cases of GCA relapse heralded by symptoms of isolated polymyalgia rheumatica (1 case versus 5 in the placebo group; P = 0.05). Conclusion. The results of this randomized, multicenter trial do not support the adjunctive use of MTX to control disease activity or to decrease the cumulative dose and toxicity of CS in patients with GCA. C1 Cleveland Clin Fdn A50, Harold C Schott Chair Rheumat & Immunol Dis, Cleveland, OH 44915 USA. Hosp Clin Barcelona, Barcelona, Spain. Johns Hopkins Univ, Sch Med, Baltimore, MD USA. Univ Paris 08, Paris, France. Pk Nicollet Med Fdn, St Louis Pk, MN USA. NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. McGill Univ, Montreal Gen Hosp, Montreal, PQ H3G 1A4, Canada. Mayo Clin, Rochester, MN USA. Marshfield Clin Fdn Med Res & Educ, Marshfield, WI USA. Univ Calif Los Angeles, Los Angeles, CA USA. Western Gen Hosp, Edinburgh EH4 2XU, Midlothian, Scotland. Ctr Rheumatol Immunol & Arthrit, Ft Lauderdale, FL USA. Rheumatol Assoc S Florida, Delray Beach, FL USA. Univ Arizona, Tucson, AZ USA. Med Univ Lubeck, D-23538 Lubeck, Germany. RP Hoffman, GS (reprint author), Cleveland Clin Fdn A50, Harold C Schott Chair Rheumat & Immunol Dis, 9500 Euclid Ave, Cleveland, OH 44915 USA. RI Easley, Kirk/K-6910-2015; OI Easley, Kirk/0000-0003-4419-2617; Luqmani, Raashid/0000-0002-4446-5841; Cid Xutgla, Maria Cinta/0000-0002-4730-0938 NR 55 TC 222 Z9 226 U1 1 U2 5 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0004-3591 J9 ARTHRITIS RHEUM JI Arthritis Rheum. PD MAY PY 2002 VL 46 IS 5 BP 1309 EP 1318 DI 10.1002/art.10262 PG 10 WC Rheumatology SC Rheumatology GA 552XU UT WOS:000175646000026 PM 12115238 ER PT J AU Gracely, RH Petzke, F Wolf, JM Clauw, DJ AF Gracely, RH Petzke, F Wolf, JM Clauw, DJ TI Functional magnetic resonance imaging evidence of augmented pain processing in fibromyalgia SO ARTHRITIS AND RHEUMATISM LA English DT Article ID POSITRON-EMISSION-TOMOGRAPHY; CEREBRAL BLOOD-FLOW; PATHOGENIC MECHANISMS; SOMATOSENSORY CORTEX; HYPNOTIC ANALGESIA; ANTERIOR CINGULATE; GENERAL-POPULATION; NEUROPATHIC PAIN; HUMAN BRAIN; HEAT PAIN AB Objective. To use functional magnetic resonance imaging (fMRI) to evaluate the pattern of cerebral activation during the application of painful pressure and determine whether this pattern is augmented in patients with fibromyalgia (FM) compared with controls. Methods. Pressure was applied to the left thumbnail beds of 16 right-handed patients with FM and 16 right-handed matched controls. Each FM patient underwent fMRI while moderately painful pressure was being applied. The functional activation patterns in FM patients were compared with those in controls, who were tested under 2 conditions: the "stimulus pressure control" condition, during which they received an amount of pressure similar to that delivered to patients, and the "subjective pain control" condition, during which the intensity of stimulation was increased to deliver a subjective level of pain similar to that experienced by patients. Results. Stimulation with adequate pressure to cause similar pain in both groups resulted in 19 regions of increased regional cerebral blood flow in healthy controls and 12 significant regions in patients. Increased fMRI signal occurred in 7 regions common to both groups, and decreased signal was observed in 1 common region. In contrast, stimulation of controls with the same amount of pressure that caused pain in patients resulted in only 2 regions of increased signal, neither of which coincided with a region of activation in patients. Statistical comparison of the patient and control groups receiving similar stimulus pressures revealed 13 regions of greater activation in the patient group. In contrast, similar stimulus pressures produced only 1 region of greater activation in the control group. Conclusion. The fact that comparable subjectively painful conditions resulted in activation patterns that were similar in patients and controls, whereas similar pressures resulted in no common regions of activation and greater effects in patients, supports the hypothesis that FM is characterized by cortical or subcortical augmentation of pain processing. C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. Georgetown Univ, Georgetown Chron Pain & Fatigue Res Ctr, Washington, DC USA. Georgetown Univ, Med Ctr, Washington, DC 20007 USA. RP Clauw, DJ (reprint author), Univ Michigan, Med Ctr, 5510 MSRB 1, Ann Arbor, MI 48109 USA. NR 52 TC 607 Z9 615 U1 1 U2 23 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0004-3591 J9 ARTHRITIS RHEUM JI Arthritis Rheum. PD MAY PY 2002 VL 46 IS 5 BP 1333 EP 1343 DI 10.1002/art.10225 PG 11 WC Rheumatology SC Rheumatology GA 552XU UT WOS:000175646000029 PM 12115241 ER PT J AU Hunt, KJ Pankow, JS Offenbacher, S Kritchevsky, SB Duncan, BB Shahar, E Sharrett, AR Heiss, G AF Hunt, KJ Pankow, JS Offenbacher, S Kritchevsky, SB Duncan, BB Shahar, E Sharrett, AR Heiss, G TI B-mode ultrasound-detected carotid artery lesions with and without acoustic shadowing and their association with markers of inflammation and endothelial activation: the atherosclerosis risk in communities study SO ATHEROSCLEROSIS LA English DT Article DE carotid arteries; ultrasonography; acoustic shadowing; inflammation; endothelial activation ID CORONARY-HEART-DISEASE; C-REACTIVE PROTEIN; ULTRAFAST COMPUTED-TOMOGRAPHY; OSTEOPONTIN MESSENGER-RNA; ADHESION MOLECULES; TISSUE CHARACTERIZATION; ATHEROMATOUS PLAQUES; ASYMPTOMATIC MEN; E-SELECTIN; CALCIFICATION AB In a cross-sectional Study of 8695 men and women free of clinical CVD, aged 45-64 years at the 1987-1989 baseline Atherosclerosis Risk in Communities (ARIC) study exam, we examined the relationship between carotid artery lesions (CALs), with and without acoustic shadowing (AS) as an index of plaque mineralization, to systemic markers of inflammation and markers of endothelial function, including endothelial adhesion molecules. A three-level variable, based on the presence of extracranial CALs and AS, identified by 13-mode ultrasound of six I cm arterial segments, defined the outcome. Among subjects without evidence of AS, after controlling for age, gender, ethnicity, study site, body mass index, hypertension. diabetes, and smoking status. CALs were associated with systemic markers of inflammation, including higher levels of fibrinogen [OR = 1.24 (95, Cl: 1.09, 1.40)] and white blood cell count [OR = 1.37 (95% CI: 1.21, 1.56)]. Among subjects with a CAL, after controlling for the above risk factors as ell as mean far wall intima-media thickness, AS was associated with higher levels of,on Willebrand factor [OR = 1.38 (95%, CI: 1.10, 1.74)], a marker of endothelial activation. Associations with endothelial adhesion molecules were inconsistent. Further studies aimed at elucidating the mechanisms of arterial mineralization are warranted. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, CVD Program, Chapel Hill, NC 27514 USA. Univ N Carolina, Sch Dent, Dept Periodontol, Chapel Hill, NC 27514 USA. Univ Tennessee, Hlth Sci Ctr, Dept Prevent Med, Memphis, TN USA. Univ Fed Rio Grande Sul, Sch Med, Dept Social Med, Porto Alegre, RS, Brazil. Univ Minnesota, Sch Publ Hlth, Div Epidemiol, Minneapolis, MN 55455 USA. NHLBI, Bethesda, MD 20892 USA. RP Heiss, G (reprint author), Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, CVD Program, 137 E Franklin St,Bank Amer,Suite 306, Chapel Hill, NC 27514 USA. FU NHLBI NIH HHS [N01-HC-55021, N01-HC-55019, N01-HC-55018, N01-HC-55016, N01-HC-55020, N01-HC-55017, N01-HC-55015, 5-T32-HL07055] NR 58 TC 15 Z9 15 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0021-9150 J9 ATHEROSCLEROSIS JI Atherosclerosis PD MAY PY 2002 VL 162 IS 1 BP 145 EP 155 AR PII S0021-9150(01)00676-1 DI 10.1016/S0021-9150(01)00676-1 PG 11 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 549RT UT WOS:000175461100017 PM 11947908 ER PT J AU Weinstein, B AF Weinstein, B TI Building the house around the plumbing SO BIOESSAYS LA English DT Article ID VASCULAR DEVELOPMENT; ENDOTHELIAL-CELLS; VASCULOGENESIS; ZEBRAFISH; DIFFERENTIATION; ANGIOGENESIS; EXPRESSION; GROWTH; MUTANT; MECHANISMS AB Signaling between growing blood vessels and the tissues that they innervate has traditionally been viewed as a one-way conversation, with organs and tissues supplying important cues for the growth and anatomical patterning of the blood vessels supplying them, but not vice-versa. Two recent papers((1,2)) now provide evidence that blood vessels can have an important role in promoting the assembly of organs and tissues. These papers show that proper formation of the pancreas((1)) and liver((2)) and induction of endocrine and hepatic cell types in these endodermal organs requires inductive signals from blood vessels. Published 2002 Wiley Periodicals, Inc. C1 NICHHD, Lab Mol Genet, Unit Vertebrate Organogenesis, NIH, Bethesda, MD 20892 USA. RP Weinstein, B (reprint author), NICHHD, Lab Mol Genet, Unit Vertebrate Organogenesis, NIH, Bldg 6B Rm 309 6 Ctr Dr, Bethesda, MD 20892 USA. NR 21 TC 2 Z9 2 U1 0 U2 1 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0265-9247 J9 BIOESSAYS JI Bioessays PD MAY PY 2002 VL 24 IS 5 BP 397 EP 400 DI 10.1002/bies.10090 PG 4 WC Biochemistry & Molecular Biology; Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics GA 548PG UT WOS:000175397100001 PM 12001261 ER PT J AU Virnik, KM Gasan, AI Maleev, VY Shestopalova, AV AF Virnik, KM Gasan, AI Maleev, VY Shestopalova, AV TI Energetics of hydration of nucleic acids of different nucleotide composition SO BIOFIZIKA LA Russian DT Article DE DNA; hydration; calorimetry; energy; Monte Carlo method ID MONTE-CARLO SIMULATION; ATOM POTENTIAL FUNCTIONS; HELIX-COIL TRANSITION; DNA HYDRATION; STRUCTURAL TRANSITIONS; B TRANSITION; ION-BINDING; WATER; STABILITY; BASES AB The energetics of hydration of natural DNA of different AT/GC content and model double-helical polyribonucleotides was studied. The results obtained by a new approach, which is based on calorimetric measurements of hydration-dehydration energy of nucleic acid-water systems at different relative humidities are presented. A correlation between the dehydration energy and the nucleotide composition of native DNA was found. The energetic characteristics of systems containing deoxynucleoside monophosphates and water clusters of different dimensions were obtained by the Monte Carlo method. The results of computer simulation correlate with the experimental calorimetric data. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Natl Acad Sci Ukraine, Inst Radiophys & Elect, UA-61085 Kharkov, Ukraine. RP Virnik, KM (reprint author), NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NR 43 TC 0 Z9 0 U1 0 U2 2 PU MEZHDUNARODNAYA KNIGA PI MOSCOW PA 39 DIMITROVA UL., 113095 MOSCOW, RUSSIA SN 0006-3029 J9 BIOFIZIKA+ JI Biofizika PD MAY-JUN PY 2002 VL 47 IS 3 BP 420 EP 426 PG 7 WC Biophysics SC Biophysics GA 560DL UT WOS:000176066900004 PM 12068595 ER PT J AU Edgerton, ME Taylor, R Powell, JI Hunter, L Simon, R Liu, ET AF Edgerton, ME Taylor, R Powell, JI Hunter, L Simon, R Liu, ET TI A bioinformatics tool to select sequences for microarray studies of mouse models of oncogenesis SO BIOINFORMATICS LA English DT Article ID GENE-EXPRESSION AB One of the challenges to the effective utilization of cDNA microarray analysis in mouse models of oncogenesis is the choice of a critical set of probes that are informative for human disease. Given the thousands of genes with a potential role in human oncogenesis and the hundreds of thousands of mouse sequences available for use as probes, selection of an informative set of mouse probes can be an overwhelming task. We have developed a web based sequence mining tool using DataBase Independent (DBI) Perl to annotate publicly available sequences. The Mouse Oncochip Design Tool uses the Mouse Genome Database (MGD) developed and maintained by the Jackson Laboratories for mouse DNA sequences. There are over 380 000 sequences in their database. The output list has been ordered to present the genes more likely to be informative in a mouse model of human cancer using a candidate set of oncogenes to order the list. Mouse sequences that represent genes that are homologous with a member of a human oncogene set are listed first. In addition it provides a set of links for information on clone source gene function. C1 Hosp Univ Penn, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA. NCI, Biometr Res Branch, Div Canc Treatment & Diag, Bethesda, MD 20892 USA. NIH, Bioinformat & Mol Anal Sect, Computat Biosci & Engn Lab, Ctr Informat Technol, Bethesda, MD 20892 USA. NCI, Div Clin Sci, Bethesda, MD 20892 USA. RP Edgerton, ME (reprint author), Hosp Univ Penn, Dept Pathol & Lab Med, 3400 Spruce St, Philadelphia, PA 19104 USA. RI Liu, Edison/C-4141-2008; OI Hunter, Lawrence/0000-0003-1455-3370 NR 9 TC 5 Z9 5 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1367-4803 J9 BIOINFORMATICS JI Bioinformatics PD MAY PY 2002 VL 18 IS 5 BP 774 EP 775 DI 10.1093/bioinformatics/18.5.774 PG 2 WC Biochemical Research Methods; Biotechnology & Applied Microbiology; Computer Science, Interdisciplinary Applications; Mathematical & Computational Biology; Statistics & Probability SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Computer Science; Mathematical & Computational Biology; Mathematics GA 562YP UT WOS:000176226800021 PM 12050079 ER PT J AU Anderson, DW Chesney, MA AF Anderson, DW Chesney, MA TI Association of perceived stress with hypopnic breathing pattern in women SO BIOLOGICAL PSYCHOLOGY LA English DT Meeting Abstract C1 NIA, NIH, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0301-0511 J9 BIOL PSYCHOL JI Biol. Psychol. PD MAY PY 2002 VL 59 IS 3 BP 264 EP 264 PG 1 WC Psychology, Biological; Behavioral Sciences; Psychology; Psychology, Experimental SC Psychology; Behavioral Sciences GA 563RY UT WOS:000176270400044 ER PT J AU Hellsten, E Bernard, DJ Owens, JW Eckhaus, M Suchy, SF Nussbaum, RL AF Hellsten, E Bernard, DJ Owens, JW Eckhaus, M Suchy, SF Nussbaum, RL TI Sertoli cell vacuolization and abnormal germ cell adhesion in mice deficient in an inositol polyphosphate 5-phosphatase SO BIOLOGY OF REPRODUCTION LA English DT Article DE epididymis; phosphatases; Sertoli cells; spermatogenesis; testis ID RECEPTOR-MEDIATED ENDOCYTOSIS; RAT TESTIS; SIGNAL-TRANSDUCTION; LOWE-SYNDROME; CADHERIN; LOCALIZATION; TRANSFERRIN; PHOSPHOINOSITIDES; 4,5-BISPHOSPHATE; CYTOSKELETON AB The dynamic nature of cellular interactions during differentiation of germ cells and their translocation from the basement membrane to the lumen of the seminiferous tubules requires the existence of complex and well-regulated cellular adhesion mechanisms in the testis. Successful migration of the developing germ cells is characterized by dynamic breakage and reformation of cadherin-containing adherens junctions between the germ cells and Sertoli cells, the polarized somatic cells of the testis that support and nourish the developing gametes. Here, we demonstrate the accumulation of abnormally swollen, actin-coated, endosome-like structures that contain intact adherens junctions and stain positive for N-cadherin and beta-catenin in the Sertoli cell cytosol of mice deficient in Inpp5b, an inositol polyphosphate 5-phosphatase. Simultaneous to the formation of these abnormal structures, developing germ cells are prematurely released from the seminiferous epithelium and sloughed into the epididymis. Our results demonstrate a role for lnpp5b in the regulation of cell adhesion in the testis and in the formation of junctional complexes with neighboring cells, and they emphasize the important and essential role of phosphoinositides in spermatogenesis. C1 NHGRI, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. NIH, Vet Resources Program, Off Director, Bethesda, MD 20892 USA. RP Nussbaum, RL (reprint author), NHGRI, Genet Dis Res Branch, NIH, 49 Convent Dr,Bldg 49,Room 4A72, Bethesda, MD 20892 USA. NR 37 TC 35 Z9 35 U1 0 U2 1 PU SOC STUDY REPRODUCTION PI MADISON PA 1603 MONROE ST, MADISON, WI 53711-2021 USA SN 0006-3363 J9 BIOL REPROD JI Biol. Reprod. PD MAY PY 2002 VL 66 IS 5 BP 1522 EP 1530 DI 10.1095/biolreprod66.5.1522 PG 9 WC Reproductive Biology SC Reproductive Biology GA 544KB UT WOS:000175155800039 PM 11967219 ER PT J AU Lee, WJ Brennan, P Boffetta, P London, SJ Benhamou, S Rannug, A To-Figueras, J Ingelman-Sundberg, M Shields, P Gaspari, L Taioli, E AF Lee, WJ Brennan, P Boffetta, P London, SJ Benhamou, S Rannug, A To-Figueras, J Ingelman-Sundberg, M Shields, P Gaspari, L Taioli, E TI Microsomal epoxide hydrolase polymorphisms and lung cancer risk: a quantitative review SO BIOMARKERS LA English DT Review DE microsomal epoxide hydrolase; polymorphism; lung cancer; smoking ID XENOBIOTIC-METABOLIZING ENZYMES; GENETIC-POLYMORPHISM; SUSCEPTIBILITY; ASSOCIATION; EXPRESSION; GENOTYPES AB To investigate the role of microsomal epoxide hydrolase (mEH) polymorphisms in the aetiology of lung cancer and to assess the interaction between mEH polymorphisms and smoking, we performed a meta-analysis of seven published studies, which included 2078 cases and 3081 controls, and a pooled analysis of eight studies (four published and four unpublished at that time) with a total of 986 cases and 1633 controls. The combined meta-analysis odds ratios (ORs) were 0.98 (95% confidence interval [CI] = 0.72-1.35) for polymorphism at amino acid 113 in exon 3 (His/His versus Tyr/Tyr genotype) and 1.00 (95% CI = 0.71-1.41) for polymorphism at amino acid 139 in exon 4 (Arg/Arg versus His/ His genotype). In the pooled analysis, we observed a significant decrease in lung cancer risk (OR = 0.70, 95% CI = 0.51=0.96) for exon 3 His/ His genotype after adjustment for age, sex, smoking and centre. The protective effect of exon 3 polymorphism seems stronger for adenocarcinoma of the lung than for other histological types. The OR for high predicted mEH activity, compared with low activity, was 1.54 (95% CI = 0.77-3.07) in the meta analysis and 1.18 (95% CI = 0.92-1.52) in the pooled analysis. We did not find a consistent modification of the carcinogenic effect of smoking according to mEH polymorphism, although the risk of lung cancer decreased among never smokers with high mEH activity and among heavy smokers with the exon 3 His/ His genotype. In conclusion, this study suggests a possible effect of mEH polymorphisms at exon 3 in modulating lung cancer. If present, this effect may vary among different populations, possibly because of interaction with genetic or environmental factors. C1 Int Agcy Res Canc, Unit Environm Canc Epidemiol, F-69008 Lyon, France. NIEHS, Res Triangle Pk, NC 27709 USA. Inst Gustave Roussy, INSERM, U521, F-94805 Villejuif, France. Natl Inst Occupat Hlth, S-17184 Solna, Sweden. Hosp Clin Barcelona, Barcelona, Spain. Karolinska Inst, Stockholm, Sweden. Georgetown Univ, Med Ctr, Washington, DC 20007 USA. Osped Maggiore, IRCCS, Milan, Italy. RP Brennan, P (reprint author), Int Agcy Res Canc, Unit Environm Canc Epidemiol, 150 Cours Albert Thomas, F-69008 Lyon, France. RI Shields, Peter/I-1644-2012; Benhamou, Simone/K-6554-2015; OI London, Stephanie/0000-0003-4911-5290 NR 18 TC 52 Z9 56 U1 1 U2 4 PU TAYLOR & FRANCIS LTD PI ABINGDON PA 4 PARK SQUARE, MILTON PARK, ABINGDON OX14 4RN, OXON, ENGLAND SN 1354-750X J9 BIOMARKERS JI Biomarkers PD MAY-JUN PY 2002 VL 7 IS 3 BP 230 EP 241 DI 10.1080/13547500210121882 PG 12 WC Biotechnology & Applied Microbiology; Toxicology SC Biotechnology & Applied Microbiology; Toxicology GA 559XH UT WOS:000176051100003 PM 12141066 ER EF