FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Hough, CD Sherman-Baust, CA Pizer, ES Montz, FJ Im, DD Rosenshein, NB Cho, KR Riggins, GJ Morin, PJ AF Hough, CD Sherman-Baust, CA Pizer, ES Montz, FJ Im, DD Rosenshein, NB Cho, KR Riggins, GJ Morin, PJ TI Large-scale serial analysis of gene expression reveals genes differentially expressed in ovarian cancer SO CANCER RESEARCH LA English DT Article ID SURFACE EPITHELIUM; MOLECULAR-CLONING; BREAST-CANCER; LYSYL OXIDASE; CARCINOMAS; CELLS; TUMORS; ORIGIN; IDENTIFICATION; PATTERNS AB Difficulties in the detection, diagnosis, and treatment of ovarian cancer result in an overall low survival rate of women with this disease. A better understanding of the pathways involved in ovarian tumorigenesis will likely provide new targets for early and effective intervention. Here, we have used serial analysis of gene expression (SAGE) to generate global gene expression profiles from various ovarian cell lines and tissues, including primary cancers, ovarian surface epithelia cells, and cystadenoma cells. The profiles were used to compare overall patterns of gene expression and to identify differentially expressed genes. We have sequenced a total of 385,000 tags, yielding 256,000 genes expressed in 10 different Libraries derived from ovarian tissues. In general, ovarian cancer cell Lines showed relatively high levels of similarity to libraries from other cancer felt lines, regardless of the tissue of origin (ovarian or colon), indicating that these lines had lost many of their tissue-specific expression patterns, In contrast, immortalized ovarian surface epithelia and ovarian cystadenoma cells showed much higher similarity to primary ovarian carcinomas than to primary colon carcinomas. Primary tissue specimens therefore appeared to be a better model for gene expression analyses. Using the expression profiles described above and stringent selection criteria, we have identified a number of genes highly differentially expressed between nontransformed ovarian epithelia and ovarian carcinomas. Some of the genes Identified are already known to be overexpressed in ovarian cancer, but several represent novel candidates. Many of the genes up-regulated in ovarian cancer represent surface or secreted proteins such as claudin-3 and -4, HE4, mucin-l, epithelial cellular adhesion molecule, and mesothelin, Interestingly, both apolipoprotein E (ApoE) and ApoJ, two proteins involved in lipid homeostasis, are among the genes highly up-regulated in ovarian cancer, Selected serial analysis of gene expression results were further validated through immunohistochemical analysis of ApoJ, claudin-3, claudin-4, and epithelial cellular adhesion molecule in archival material. These experiments provided additional evidence of the relevance of our findings in vivo. The publicly available expression data reported here should stimulate and aid further research in the field of ovarian cancer. C1 NIA, Gerontol Res Ctr, Biol Chem Lab, NIH, Baltimore, MD 21224 USA. Johns Hopkins Med Inst, Dept Pathol, Baltimore, MD 21287 USA. Johns Hopkins Med Inst, Dept Gynecol, Baltimore, MD 21287 USA. Mercy Med Ctr, Gynecol Oncol Ctr, Baltimore, MD 21202 USA. Univ Michigan, Sch Med, Dept Pathol, Ann Arbor, MI 48109 USA. Duke Univ, Med Ctr, Dept Pathol, Durham, NC 27710 USA. RP Morin, PJ (reprint author), NIA, Gerontol Res Ctr, Biol Chem Lab, NIH, 5600 Nathan Shock St, Baltimore, MD 21224 USA. NR 34 TC 367 Z9 384 U1 1 U2 19 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 NOV 15 PY 2000 VL 60 IS 22 BP 6281 EP 6287 PG 7 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600013 PM 11103784 ER PT J AU Paweletz, CP Ornstein, DK Roth, MJ Bichsel, VE Gillespie, JW Calvert, VS Vocke, CD Hewitt, SM Duray, PH Herring, J Wang, QH Hu, N Linehan, WM Taylor, PR Liotta, LA Emmert-Buck, MR Petricoin, EF AF Paweletz, CP Ornstein, DK Roth, MJ Bichsel, VE Gillespie, JW Calvert, VS Vocke, CD Hewitt, SM Duray, PH Herring, J Wang, QH Hu, N Linehan, WM Taylor, PR Liotta, LA Emmert-Buck, MR Petricoin, EF TI Loss of annexin 1 correlates with early onset of tumorigenesis in esophageal and prostate carcinoma SO CANCER RESEARCH LA English DT Article ID LASER-CAPTURE MICRODISSECTION; GENE-EXPRESSION; LIPOCORTIN-I; CANCER; DIFFERENTIATION; INDUCTION; CELLS AB Annexin I protein expression was evaluated in patient-matched longitudinal study sets of laser capture microdissected normal, premalignant, and invasive epithelium from human esophageal squamous cell cancer and prostatic adenocarcinoma, In 25 esophageal cases (20 by Western blot and 5 by immunohtstochemistry) and 17 prostate cases (3 by Western blot and 14 by immunohistochemistry), both tumor types showed either complete loss or a dramatic reduction in the level of annexin I protein expression compared with patient-matched normal epithelium (P less than or equal to 0,05), Moreover, by using Western blot analysis of laser capture microdissected, patient-matched longitudinal study sets of both tumor types, the loss of protein expression occurred in premalignant lesions. Concordance of this result with immunohistochemical analysis suggests that annexin I may be an essential component for maintenance of the normal epithelial phenotype. Additional studies investigating the mechanism(s) and functional consequences of annexin I protein loss in tumor cells are warranted. C1 US FDA, Ctr Biol Evaluat & Res, Div Therapeut Prod, Bethesda, MD 20892 USA. Georgetown Univ, Dept Chem, Washington, DC 20057 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. NCI, Urol Oncol Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathogenet Unit, Pathol Lab, NIH, Bethesda, MD 20892 USA. NCI, Canc Prevent Studies Branch, NIH, Bethesda, MD 20892 USA. NCI, Canc Genome Anat Project, Off Director, NIH, Bethesda, MD 20892 USA. Natl Naval Med Ctr, Bethesda, MD 20892 USA. Shanxi Canc Hosp, Pathol Lab, Taiyuan 030013, Peoples R China. RP Petricoin, EF (reprint author), US FDA, Ctr Biol Evaluat & Res, Div Therapeut Prod, Bethesda, MD 20892 USA. OI Hewitt, Stephen/0000-0001-8283-1788 NR 20 TC 162 Z9 184 U1 0 U2 3 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 NOV 15 PY 2000 VL 60 IS 22 BP 6293 EP 6297 PG 5 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600015 PM 11103786 ER PT J AU Rutberg, SE Adams, TL Glick, A Bonovich, MT Vinson, C Yuspa, SH AF Rutberg, SE Adams, TL Glick, A Bonovich, MT Vinson, C Yuspa, SH TI Activator protein 1 transcription factors are fundamental to v-ras(Ha)-induced changes in gene expression in neoplastic keratinocytes SO CANCER RESEARCH LA English DT Article ID MOUSE SKIN CARCINOGENESIS; C-FOS; REGULATORY ELEMENT; DNA-BINDING; CELL-LINES; ONCOGENE ACTIVATION; TISSUE-SPECIFICITY; TRANSGENIC MICE; EPIDERMAL-CELLS; HUMAN KERATIN-1 AB The induction of mouse skin papillomas by initiation-promotion protocols is associated with aberrant expression of epithelial markers in the tumor mass. Similarly, initiation of mouse keratinocytes with a retrovirus encoding the v-ras(Ha) gene (v-ras(Ha) keratinocytes) causes characteristic alterations of epidermal gene expression (A, A. Dlugosz et al., Cancer Res., 54: 6413-6420, 1994), Because activator protein 1 (AP-1) proteins are likely targets of Ras activation, we have examined the role of AP-1 factors in v-ras(Ha) keratinocytes. Introduction of v-ras(Ha) into keratinocytes upregulates c-Fos, Delta Fos B, and Fra-1 transcripts and protein levels in nuclear extracts. The expression of Jun proteins is not significantly altered in v-ras(Ha) keratinocytes, Transduction of cells with v-ras(Ha) results in increased AP-1-dependent transcriptional activity, which is also simulated by transfection of keratinocytes with either c-Fos or Delta Fos B but not Fra-1, suggesting that the up-regulation of c-Fos and Delta Fos B contributes to this effect. To explore the role of AP-I proteins in regulating keratinocyte markers in v-rasHa keratinocytes, we blocked the binding of AP-I proteins to DNA by infecting keratinocytes with an adenovirus encoding a dominant-negative Fos mutant (A-FOS), A-FOS replaces endogenous Fos proteins in the formation of heterodimers with Jun family members and thus prevents the AP-1 transcription factor from binding to DNA, In v-ras(Ha) keratinocytes, the A-FOS virus reversed the suppression of keratins 1 and 10 transcripts and protein, which is characteristically seen in tumors and v-ras(Ha) keratinocytes, A-FOS also increased protein levels but reduced transcripts for the late marker, loricrin, a component of the cornified envelope. These findings indicate that AP-1 proteins are involved in the changes in gene expression that define the v-ras(Ha) phenotype in mouse keratinocytes. C1 NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bethesda, MD 20892 USA. NCI, Lab Metab, Bethesda, MD 20892 USA. RP Yuspa, SH (reprint author), NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bldg 37,Room 3B25,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 67 TC 18 Z9 19 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 15 PY 2000 VL 60 IS 22 BP 6332 EP 6338 PG 7 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600023 PM 11103794 ER PT J AU Ratnasinghe, D Tangrea, JA Andersen, MR Barrett, MJ Virtamo, J Taylor, PR Albanes, D AF Ratnasinghe, D Tangrea, JA Andersen, MR Barrett, MJ Virtamo, J Taylor, PR Albanes, D TI Glutathione peroxidase codon 198 polymorphism variant increases lung cancer risk SO CANCER RESEARCH LA English DT Article ID ALPHA-TOCOPHEROL; BETA-CAROTENE; SELENIUM; GENE; EXPRESSION; LINE AB Human cellular glutathione peroxidase 1 (hGPX1) is a selenium-dependent enzyme that participates in the detoxification of hydrogen peroxide and a wide range of organic peroxides. We conducted a case-control study nested within the alpha -Tocopherol, beta -Carotene Cancer Prevention Study cohort to evaluate the association between the proline to leucine polymorphism at codon 198 of hGPX1 and lung cancer risk. Cases (n = 315) were matched to controls on age (+/-5 years), intervention group, and study clinic using incidence density sampling in a 1:1 ratio, The prevalence of the hGPX1 Pro198leu variant allele was 58% for controls and 71% Fur cases (P < 0.001). Using conditional logistic regression, we found a significant association between hGPX1 genotype and lung cancer risk. The odds ratio for heterozygotes was 1.8 (95% confidence interval, 1.2-2.8) and 2.3 (95% confidence interval, 1.3-3.8) for homozygous variants compared to wild-type individuals. Due to its high prevalence, the hGPX1 variant may contribute significantly to lung cancer risk among Caucasians but not among ethnic Chinese who do not exhibit this polymorphism. C1 NCI, Div Clin Sci, Canc Prevent Studies Branch, NIH, Bethesda, MD 20892 USA. New Chem Entities Inc, Thetagen Div, Bothell, WA 98011 USA. Informat Management Serv Inc, Silver Spring, MD 20904 USA. Natl Publ Hlth Inst, SF-00300 Helsinki, Finland. RP Ratnasinghe, D (reprint author), NCI, Div Clin Sci, Canc Prevent Studies Branch, NIH, 6006 Execut Blvd,MSC 7058, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 FU NCI NIH HHS [N01-CN45165] NR 20 TC 167 Z9 168 U1 0 U2 3 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 NOV 15 PY 2000 VL 60 IS 22 BP 6381 EP 6383 PG 3 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600030 PM 11103801 ER PT J AU Mononen, N Syrjakoski, K Matikainen, M Tammela, TLJ Schleutker, J Kallioniemi, OP Trapman, J Koivisto, PA AF Mononen, N Syrjakoski, K Matikainen, M Tammela, TLJ Schleutker, J Kallioniemi, OP Trapman, J Koivisto, PA TI Two percent of Finnish prostate cancer patients have a germ-line mutation in the hormone-binding domain of the androgen receptor gene SO CANCER RESEARCH LA English DT Article ID VITAMIN-D-RECEPTOR; SUSCEPTIBILITY LOCUS; CAG REPEAT; RISK; CHROMOSOME; BRCA1; POLYMORPHISMS; MEN AB Mutations of the androgen receptor (AR) gene have been reported in prostate cancer, usually from tumor tissue specimens from late-stage, androgen-independent cancer. Occasionally, germ-line mutations have been found, but a link between AR mutations and predisposition to human prostate cancer has not been firmly established. Recently, two independent studies reported the same germ-line mutation at codon 726 in exon E (CGC to CTC) in two apparently unrelated Finnish prostate cancer patients. This arginine to leucine substitution was reported to alter the transactivational specificity of the AR protein. In the present study, the R726L mutation was analyzed by allele-specific oligohybridization in DNA specimens from 418 consecutive prostate cancer patients who reported a negative family history (sporadic group) and from 106 patients with a positive family history (hereditary group), The population frequency of the R726L mutation in blood donors was 3 of 900 (0.33%), In contrast, eight (1.91%) mutations (odds ratio = 5.8; P = 0.006) were found in the sporadic group, and two (1.89%) mutations were found in the hereditary group (odds ratio = 5.8; P = 0.09), Suggestive evidence of the segregation of the mutation with prostate cancer was seen in these two families. The present study indicates that the R726L substitution in the AR may confer an up to 6-fold increased risk of prostate cancer and may contribute to cancer development in up to 2% of Finnish prostate cancer patients. These results warrant additional large-scale studies of the significance of rare mutations and polymorphisms in candidate genes along the androgen signaling pathway as risk factors for prostate cancer. C1 Tampere Univ Hosp, Canc Genet Lab, FIN-33521 Tampere, Finland. Univ Tampere, Inst Med Technol, Canc Genet Lab, FIN-33521 Tampere, Finland. Tampere Univ Hosp, Dept Clin Genet, FIN-33521 Tampere, Finland. Tampere Univ Hosp, Div Urol, FIN-33521 Tampere, Finland. Univ Tampere, Sch Med, FIN-33521 Tampere, Finland. NIH, Natl Ctr Human Genome Res, Canc Genet Lab, Bethesda, MD 20892 USA. Erasmus Univ, Dept Pathol, NL-3000 DR Rotterdam, Netherlands. RP Koivisto, PA (reprint author), Tampere Univ Hosp, Canc Genet Lab, POB 2000, FIN-33521 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 FU NHGRI NIH HHS [N01-HG-55389] NR 30 TC 50 Z9 52 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 15 PY 2000 VL 60 IS 22 BP 6479 EP 6481 PG 3 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600045 PM 11103816 ER PT J AU Jiang, F Desper, R Papadimitriou, CH Schaffer, AA Kallioniemi, OP Richter, J Schraml, P Sauter, G Mihatsch, MJ Moch, H AF Jiang, F Desper, R Papadimitriou, CH Schaffer, AA Kallioniemi, OP Richter, J Schraml, P Sauter, G Mihatsch, MJ Moch, H TI Construction of evolutionary tree models for renal cell carcinoma from comparative genomic hybridization data SO CANCER RESEARCH LA English DT Article ID SEQUENCE COPY NUMBER; INTRATUMORAL HETEROGENEITY; INTERPHASE CYTOGENETICS; CHROMOSOMAL IMBALANCES; RETINOBLASTOMA GENE; CORTICAL NEOPLASMS; BLADDER-CANCER; SOLID TUMORS; ALLELIC LOSS; SHORT ARM AB Renal cell carcinoma is characterized by an accumulation of complex chromosomal alterations during tumor progression. Chromosome 3p deletions are known to occur early in the carcinogenesis, but the nature of subsequent events, their interrelationships, and their sequence is poorly understood, as one usually only obtains a single "view'" of the dynamic process of tumor development in a particular cancer patient. To address this limitation, we used comparative genomic hybridization analysis in combination with a distance-based and a branching-tree method to search for tree models of the oncogenesis process of 116 conventional (clear cell) renal carcinomas. This provides a means to analyze and model cancer development processes based on a more dynamic model, including the presence of multiple pathways, as compared with the fixed linear model first proposed by Vogelstein st al. (N. Engl. J. Med., 319: 525-532, 1988) for colorectal cancer. The most common DNA losses involved 3p (61%), 4q (50%), 6q (40%), 9p (35%), 13q (37%), and Xq (21%). The most common gains were seen at chromosome 17p and 17q (20%), The tree model derived from the distance-based method is consistent with the established theory that -3p is an important early event in conventional (clear cell) renal cancer and supports the prediction made from the branching tree that -4q is another important early event. Both tree models suggest that there may be two groups of clear cell renal cancers: one characterized by -6q, +17q, and +17p, and another by -9p, -13q, and -18q. Putative prognostic parameters were -9p and -13q. The distance-based tree clarifies that -8p (present in 12% of tumors) is a late event, largely independent of other events. In summary, tree modeling of comparative genomic hybridization data provided new information on the interrelationships of genetic changes in renal cancer and their possible order, as well as a clustering of these events. Using tree analysis, one can derive a more in-depth understanding of the renal cancer development process than is possible by simply focusing on the frequencies of genetic events in a given cancer type. C1 Univ Basel, Inst Pathol, CH-4003 Basel, Switzerland. Deutsch Krebsforschungszentrum, Abt Theoret Bioinformat, D-69120 Heidelberg, Germany. Univ Calif Berkeley, Dept Elect Engn & Comp Sci, Berkeley, CA 94720 USA. NIH, Natl Ctr Biotechnol Informat, Computat Biol Branch, Bethesda, MD 20894 USA. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. RP Moch, H (reprint author), Univ Basel, Inst Pathol, Schonbeinstr 40, CH-4003 Basel, Switzerland. RI Kallioniemi, Olli/H-5111-2011; Schaffer, Alejandro/F-2902-2012; Kallioniemi, Olli/H-4738-2012 OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332 NR 64 TC 83 Z9 88 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 15 PY 2000 VL 60 IS 22 BP 6503 EP 6509 PG 7 WC Oncology SC Oncology GA 375ZD UT WOS:000165432600049 PM 11103820 ER PT J AU Park, HC Kim, CH Bae, YK Yee, SY Kim, SH Hong, SK Shin, J Yoo, KW Hibi, M Hirano, T Miki, N Chitnis, AB Huh, TL AF Park, HC Kim, CH Bae, YK Yee, SY Kim, SH Hong, SK Shin, J Yoo, KW Hibi, M Hirano, T Miki, N Chitnis, AB Huh, TL TI Analysis of upstream elements in the HuC promoter leads to the establishment of transgenic zebrafish with fluorescent neurons SO DEVELOPMENTAL BIOLOGY LA English DT Article DE HuC; elav; zebrafish; neuron; promoter; Myt1; E box; transgenic; mind bomb ID AU-RICH ELEMENTS; DROSOPHILA-MELANOGASTER; MESSENGER-RNA; EMBRYONIC-DEVELOPMENT; PRIMARY NEUROGENESIS; BINDING PROTEINS; REPORTER GENE; LOCUS ELAV; DELTA-GENE; CBF/NF-Y AB HuC encodes an RNA binding protein homologous to Drosophila elav that serves as an excellent early marker for differentiating neurons. We have characterized the promoter of the zebrafish HuC gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos. We determined that 2.8 kb of the 5'-flanking sequence is sufficient to restrict GFP gene expression to neurons. The core promoter spans 251 base pairs and contains a CCAAT box and one SP1 sequence but no TATA box is present near the transcription start site. A putative MyT1 binding site and at least 17 E-box sequences are necessary to maintain the neuronal specificity of HuC expression. Interestingly, sequential removal of the putative MyT1 binding site and 14 distal E boxes does not appear to abolish neuronal expression; rather, it leads to a progressive expansion of GFP expression into muscle cells. Further removal of the three proximal E boxes eliminates neuronal and muscle specificity of GFP expression and leads to ubiquitous expression of GFP in the whole body. Identification of key components of the HuC promoter has led to the establishment of a stable zebrafish transgenic line (HuC-GFP) in which GFP is expressed specifically in neurons. We crossed mind bomb (mib) fish with this line to visualize their neurogenic phenotype in live mib(-/-) mutant embryos. This cross illustrates how HuC-GFP fish could be used in the future to identify and analyze zebrafish mutants with an aberrant pattern of early neurons. (C) 2000 Academic Press. C1 Kyungpook Natl Univ, Coll Nat Sci, Dept Genet Engn, Taegu 702701, South Korea. NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. Osaka Univ, Sch Med, Dept Pharmacol, Suita, Osaka 565, Japan. RP Huh, TL (reprint author), Kyungpook Natl Univ, Coll Nat Sci, Dept Genet Engn, Taegu 702701, South Korea. RI Hirano, Toshio/C-8194-2009 NR 54 TC 210 Z9 212 U1 1 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD NOV 15 PY 2000 VL 227 IS 2 BP 279 EP 293 DI 10.1006/dbio.2000.9898 PG 15 WC Developmental Biology SC Developmental Biology GA 374UP UT WOS:000165363000004 PM 11071755 ER PT J AU Kramer, PR Wray, S AF Kramer, PR Wray, S TI Midline nasal tissue influences nestin expression in nasal-placode-derived luteinizing hormone-releasing hormone neurons during development SO DEVELOPMENTAL BIOLOGY LA English DT Article DE LHRH neurons; differentiation; gene expression; olfactory receptor neurons; lineage; nestin; Mash-1; GnRH ID CENTRAL-NERVOUS-SYSTEM; OLFACTORY PLACODE; MESSENGER-RNA; EXPLANT CULTURES; PRECURSOR CELLS; MOUSE EMBRYO; STEM-CELL; IN-VITRO; MIGRATION; PROTEIN AB Neurons differentiating into the luteinizing hormone-releasing hormone (LHRH) neuroendocrine phenotype are derived from the nasal placode. Cells within the vomeronasal organ anlage that turn on LHRH gene and peptide expression subsequently migrate into the forebrain where they influence reproductive function. The molecular and cellular cues regulating differentiation and migration of these cells are unknown. Discovery of developmental markers can indicate proteins directing or associated with differentiation. Analysis of such markers after manipulation of external cues can elucidate important extracellular differentiation signals. Embryonic LHRH neurons were examined in vivo for Mash-1 and nestin, two factors that delineate precursor populations in PNS and forebrain CNS cells. Nestin, but not Mash-1, was detected in early expressing LHRH cells in the vomeronasal organ anlage. These results were duplicated in LHRH neurons maintained in vitro in nasal explants. Such LHRH cells expressed nestin mRNA but not Mash-1 mRNA and were also negative for three other olfactory epithelial developmental transcription factors, Math4A, Math4C/neurogenin1, and NeuroD mRNA. Experimental manipulation of nasal explants revealed dual expression of nestin protein and LHRH in cells proximal to the vomeronasal organ anlage that was dependent upon midline cartilaginous/mesenchymal tissues. Prolonged nestin expression in LHRH cells after midline removal is consistent with nasal midline tissues modulating differentiation of LHRH neurons from the nasal placode. C1 NINDS, Cellular & Dev Neurobiol Sect, NIH, Bethesda, MD 20892 USA. RP NINDS, Cellular & Dev Neurobiol Sect, NIH, Bldg 36,Room 5A-25, Bethesda, MD 20892 USA. EM swray@codon.nih.gov OI Kramer, Phillip/0000-0003-0117-542X; wray, susan/0000-0001-7670-3915 NR 44 TC 24 Z9 25 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 0012-1606 EI 1095-564X J9 DEV BIOL JI Dev. Biol. PD NOV 15 PY 2000 VL 227 IS 2 BP 343 EP 357 DI 10.1006/dbio.2000.9896 PG 15 WC Developmental Biology SC Developmental Biology GA 374UP UT WOS:000165363000008 PM 11071759 ER PT J AU Chin, AJ Tsang, M Weinberg, ES AF Chin, AJ Tsang, M Weinberg, ES TI Heart and gut chiralities are controlled independently from initial heart position in the developing zebrafish SO DEVELOPMENTAL BIOLOGY LA English DT Article DE left-right axis; heart looping; gut chirality; heterotaxy; zebrafish ID LEFT-RIGHT ASYMMETRY; LEFT-RIGHT AXIS; MUTATIONS AFFECTING DEVELOPMENT; MIDBRAIN-HINDBRAIN BOUNDARY; NODAL-RELATED GENES; HOMEOBOX GENE; DANIO-RERIO; SIGNALING PATHWAY; CARDIOVASCULAR-SYSTEM; EXPRESSION PATTERN AB A fundamental problem in developmental biology is how left-right (LR) asymmetry is generated, both on the whole organism level and at the level of an individual organ or structure. To investigate the relationship of organ sidedness to organ chirality, we examined 12 zebrafish mutants for initial heart tube position and later heart looping direction (chirality). Anomalous initial heart position was found in seven mutants, which also demonstrated loss of normal LR asymmetry in lateral plate mesoderm (LPM) antivin/lefty-1 and Pitx2 expression. Those with a relatively normal notochord (cyc(b16), din, and spt) displayed a predictive correlation between initial heart position and heart chirality, whereas initial heart position and heart chirality were independently randomized in those with a defective notochord (flh, bet, ntl, and mom). The predictability of heart chirality in spt, din, and b16 embryos, even in the absence of normal antivin/lefty-1 and Pitx2 expression, strongly suggests that heart chirality is controlled by a process distinct from that which controls appropriate left-sided LPM expression of antivin-Pitx2 signaling pathway molecules. In addition, there was correlation of initial heart position with gut chirality (and also between heart chirality and gut chirality) in the first class of mutants with normal notochord, but not in the second class, which appears to model human heterotaxy syndrome. (C) 2000 Academic Press. C1 Univ Penn, Dept Biol, Philadelphia, PA 19104 USA. Univ Penn, Sch Med, Dept Pediat, Div Cardiol, Philadelphia, PA 19104 USA. NICHHD, Mol Genet Lab, NIH, Bethesda, MD 20892 USA. RP Chin, AJ (reprint author), Childrens Hosp Philadelphia, Cardiac Ctr, Philadelphia, PA 19104 USA. RI TSANG, Michael/E-2758-2013; Tsang, Michael/I-9305-2014; OI TSANG, Michael/0000-0001-6384-2422; Tsang, Michael/0000-0001-7123-0063; Chin, Alvin/0000-0002-0417-8653 NR 94 TC 43 Z9 45 U1 1 U2 6 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD NOV 15 PY 2000 VL 227 IS 2 BP 403 EP 421 DI 10.1006/dbio.2000.9924 PG 19 WC Developmental Biology SC Developmental Biology GA 374UP UT WOS:000165363000012 PM 11071763 ER PT J AU Junop, MS Modesti, M Guarne, A Ghirlando, R Gellert, M Yang, W AF Junop, MS Modesti, M Guarne, A Ghirlando, R Gellert, M Yang, W TI Crystal structure of the Xrcc4 DNA repair protein and implications for end joining SO EMBO JOURNAL LA English DT Article DE DNA ligase IV; NHEJ; structure; tetramer; Xrcc4 ID STRAND BREAK REPAIR; LIGASE-IV; V(D)J RECOMBINATION; CHROMOSOMES SMC; MAMMALIAN-CELLS; MAINTENANCE; LETHALITY; INTERACTS; DOMAINS; BINDING AB XRCC4 is essential for carrying out non-homologous DNA end joining (NHEJ) in all eukaryotes and, in particular, V(D)J recombination in vertebrates. Xrcc4 protein forms a complex with DNA ligase IV that rejoins two DNA ends in the last step of V(D)J recombination and NHEJ to repair double strand breaks. XRCC4-defective cells are extremely sensitive to ionizing radiation, and disruption of the XRCC4 gene results in embryonic lethality in mice. Here we report the crystal structure of a functional fragment of Xrcc4 at 2.7 A resolution. Xrcc4 protein forms a strikingly elongated dumb-bell-like tetramer. Each of the N-terminal globular head domains consists of a beta -sandwich and a potentially DNA-binding helix-turn-helix motif. The C-terminal stalk comprising a single alpha -helix >120 Angstrom in length is partly incorporated into a four-helix bundle in the Xrcc4 tetramer and partly involved in interacting with ligase IV. The Xrcc4 structure suggests a possible mode of coupling. ligase IV association with DNA binding for effective ligation of DNA ends. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Yang, W (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RI Ghirlando, Rodolfo/A-8880-2009; Yang, Wei/D-4926-2011; Junop, Murray/E-4160-2015 OI Yang, Wei/0000-0002-3591-2195; Junop, Murray/0000-0001-6676-5717 NR 35 TC 114 Z9 116 U1 1 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 15 PY 2000 VL 19 IS 22 BP 5962 EP 5970 DI 10.1093/emboj/19.22.5962 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 378AK UT WOS:000165558900003 PM 11080143 ER PT J AU Pile, LA Wassarman, DA AF Pile, LA Wassarman, DA TI Chromosomal localization links the SIN3-RPD3 complex to the regulation of chromatin condensation, histone acetylation and gene expression SO EMBO JOURNAL LA English DT Article DE chromatin; deacetylation; RPD3; SIN3; transcription ID RNA-POLYMERASE-II; DROSOPHILA POLYTENE CHROMOSOMES; SALIVARY-GLAND CHROMOSOMES; TRANSCRIPTIONAL REPRESSION; SACCHAROMYCES-CEREVISIAE; ECDYSONE RECEPTOR; IN-VIVO; DEACETYLASE COMPLEX; NUCLEOSOMAL DNA; H3 ACETYLATION AB Acetylation of core histone N-terminal tails influences chromatin condensation and transcription. To examine how the SIN3-RPD3 deacetylase complex contributes to these events in vivo, we examined binding of SIN3 and RPD3 to Drosophila salivary gland polytene chromosomes. The binding patterns of SIN3 and RPD3 were highly coincident, suggesting that the SIN3-RPD3 complex is the most abundant chromatin-bound RPD3 complex in salivary gland cells. SIN3-RPD3 binding was restricted to less condensed, hypo-acetylated euchromatic interbands and was absent from moderately condensed, hyperacetylated euchromatic bands and highly condensed, differentially acetylated centric heterochromatin. Consistent with its demonstrated role in transcriptional repression, SIN3-RPD3 did not co-localize with RNA polymerase II. Chromatin binding of the complex, mediated by SMRTER, decreased upon ecdysone-induced transcriptional activation but was restored when transcription was reduced. These results implicate SIN3-RPD3 in maintaining histone acetylation levels or patterns within less condensed chromatin domains and suggest that SIN3-RPD3 activity is required, in the absence of an activation signal, to repress transcription of particular genes within transcriptionally active chromatin domains. C1 NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Wassarman, DA (reprint author), NICHHD, Cell Biol & Metab Branch, NIH, Bldg 18T,Room 101, Bethesda, MD 20892 USA. NR 85 TC 61 Z9 61 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 15 PY 2000 VL 19 IS 22 BP 6131 EP 6140 DI 10.1093/emboj/19.22.6131 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 378AK UT WOS:000165558900019 PM 11080159 ER PT J AU Hofmann, A Zdanov, A Genschik, P Ruvinov, S Filipowicz, W Wlodawer, A AF Hofmann, A Zdanov, A Genschik, P Ruvinov, S Filipowicz, W Wlodawer, A TI Structure and mechanism of activity of the cyclic phosphodiesterase of Appr > p, a product of the tRNA splicing reaction SO EMBO JOURNAL LA English DT Article DE ADP-ribose 1 '',2 ''-cyclic phosphate; Arabidopsis; 2 ',3 '-cyclic nucleotide phosphodiesterase; tRNA splicing; X-ray crystallography ID 2',3'-CYCLIC NUCLEOTIDE 3'-PHOSPHODIESTERASE; RIBOSE 1'',2''-CYCLIC PHOSPHATE; ELECTRON-DENSITY MAPS; TRANSFER-RNA LIGASE; ESCHERICHIA-COLI; SACCHAROMYCES-CEREVISIAE; CRYSTAL-STRUCTURES; PROTEIN STRUCTURES; HEAVY-ATOM; WHEAT-GERM AB The crystal structure of the cyclic phosphodiesterase (CPDase) from Arabidopsis thaliana, an enzyme involved in the tRNA splicing pathway, was determined at 2.5 Angstrom resolution. CPDase hydrolyzes ADP-ribose 1 " ,2 " -cyclic phosphate (Appr>p), a product of the tRNA splicing reaction, to the monoester ADP-ribose 1 " -phosphate (Appr-1 "p). The 181 amino acid protein shows a novel, bilobal arrangement of two alpha beta modules. Each lobe consists of two alpha -helices on the outer side of the molecule, framing a three- or four-stranded antiparallel beta -sheet in the core of the protein. The active site is formed at the interface of the two beta -sheets in a water-filled cavity involving residues from two H-X-T/S-X motifs. This previously noticed motif participates in coordination of a sulfate ion, A solvent-exposed surface loop (residues 100-115) is very likely to play a flap-like role, opening and closing the active site. Based on the crystal structure and on recent mutagenesis studies of a homologous CPDase from Saccharomyces cerevisiae, we propose an enzymatic mechanism that employs the nucleophilic attack of a water molecule activated by one of the active site histidines. C1 NCI, Macromol Crystallog Lab, Program Struct Biol, Frederick, MD 21702 USA. NCI, Div Basic Sci, Biochem Lab, NIH, Bethesda, MD 20892 USA. Friedrich Miescher Inst, CH-4058 Basel, Switzerland. RP Hofmann, A (reprint author), NCI, Macromol Crystallog Lab, Program Struct Biol, Frederick, MD 21702 USA. RI Hofmann, Andreas/B-9515-2008 OI Hofmann, Andreas/0000-0003-4408-5467 NR 53 TC 41 Z9 42 U1 1 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 15 PY 2000 VL 19 IS 22 BP 6207 EP 6217 DI 10.1093/emboj/19.22.6207 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 378AK UT WOS:000165558900026 PM 11080166 ER PT J AU Brendler, T Sawitzke, J Sergueev, K Austin, S AF Brendler, T Sawitzke, J Sergueev, K Austin, S TI A case for sliding SeqA tracts at anchored replication forks during Escherichia coli chromosome replication and segregation SO EMBO JOURNAL LA English DT Article DE Escherichia coli; hemi-methylation; nucleoid segregation; replication forks; SeqA protein ID PROTEIN; INITIATION; ORIGIN; BINDS; SEQUESTRATION; LOCALIZATION; COMPLEXES; MIGRATION; MEMBRANES; SEQUENCE AB SeqA is an Escherichia coli DNA-binding protein that acts at replication origins and controls DNA replication. However, binding is not exclusive to origins. Many fragments containing two or more hemi-methylated GATC sequences bind efficiently. Binding was optimal when two such sequences were closely apposed or up to 31 bases apart on the same face of the DNA helix. Binding studies suggest that neighboring bound proteins contact each other to form a complex with the intervening DNA looped out. There are many potential binding sites distributed around the E.coli chromosome. As replication produces a transient wave of hemi-methylation, tracts of SeqA binding are likely to associate with each fork as replication progresses. The number and positions of green fluorescent protein-SeqA foci seen in living cells suggest that they correspond to these tracts, and that the forks are tethered to planes of cell division, SeqA may help to tether the forks or to organize newly replicated DNA into a structure that aids DNA to segregate away from the replication machinery. C1 NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Gene Regulat & Chromosome Biol Lab, Frederick, MD 21702 USA. RP Austin, S (reprint author), NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Gene Regulat & Chromosome Biol Lab, Frederick, MD 21702 USA. NR 23 TC 93 Z9 95 U1 1 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 15 PY 2000 VL 19 IS 22 BP 6249 EP 6258 DI 10.1093/emboj/19.22.6249 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 378AK UT WOS:000165558900030 PM 11080170 ER PT J AU Alper, O De Santis, ML Strombreg, K Hacker, NF Cho-Chung, YS Salomon, DS AF Alper, O De Santis, ML Strombreg, K Hacker, NF Cho-Chung, YS Salomon, DS TI Anti-sense suppression of epidermal growth factor receptor expression alters cellular proliferation, cell-adhesion and tumorigenicity in ovarian cancer cells SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID E-CADHERIN EXPRESSION; PROSTATE CARCINOMA-CELLS; EGF SUPERGENE FAMILY; BETA-CATENIN; PROTEIN INTERACTIONS; SIGNAL-TRANSDUCTION; MEDIATED ADHESION; EPITHELIAL-CELLS; BREAST-CANCER; IN-VIVO AB Over-expression of epidermal growth factor receptor (EGFR) in ovarian cancer has been well documented. Human NIH:OVCAR-8 ovarian carcinoma cells were transfected with an expression vector containing the anti-sense orientation of truncated human EGFR cDNA. EGFR anti-sense overexpression resulted in decreased EGFR protein and mRNA expression, cell proliferation and tumor formation in nude mice. In accordance with the reduced levels of EGFR in EGFR anti-sense-expressing cells, tyrosine phosphorylation of EGFR was decreased compared to untransfected parental cells treated with EGF. In EGFR anti-sense-transfected cells, expression of erbB-3, but not erbB-2, was increased. In addition, basal and heregulin-beta I-stimulated tyrosine phosphorylation of erbB-3 was higher in EGFR anti-sense vector-transfected cells. A morphological alteration in EGFR anti-sense gene-expressing cells was correlated with a decrease in the expression of E-cadherin, alpha -catenin and, to a lesser extent, beta -catenin. Changes in the expression of these proteins were associated with a reduction in complex formation among E-cadherin, beta -catenin and alpha -catenin and between beta -catenin and EGFR in EGFR anti-sense-expressing cells compared to sense-transfected control cells. These results demonstrate that EGFR expression in ovarian carcinoma cells regulates expression of cell adhesion proteins that may enhance cell growth and invasiveness. Published 2000 Wiley-Liss, Inc.(dagger). C1 NCI, Tumor Immunol & Biol Lab, Tumor Growth Factor Sect, NIH, Bethesda, MD 20892 USA. NCI, Tumor Immunol & Biol Lab, Cellular Biochem Sect, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Div Therapeut Prot, Bethesda, MD USA. Royal Hosp Women, Gynaecol Canc Ctr, Randwick, NSW, Australia. Walter Reed Army Inst Res, Dept Mol Pathol, Silver Spring, MD USA. RP NCI, Tumor Immunol & Biol Lab, Tumor Growth Factor Sect, NIH, 9200 Rockville Pike,Bldg 10,Room 5B39, Bethesda, MD 20892 USA. EM davetgfa@helix.nih.gov NR 52 TC 45 Z9 48 U1 0 U2 3 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0020-7136 EI 1097-0215 J9 INT J CANCER JI Int. J. Cancer PD NOV 15 PY 2000 VL 88 IS 4 BP 566 EP 574 DI 10.1002/1097-0215(20001115)88:4<566::AID-IJC8>3.0.CO;2-D PG 9 WC Oncology SC Oncology GA 368TW UT WOS:000090134500008 PM 11058872 ER PT J AU Hodgson, L Kohn, EC Dong, C AF Hodgson, L Kohn, EC Dong, C TI Extracellular lipid-mediated signaling in tumor-cell activation and pseudopod protrusion SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID LYSOPHOSPHATIDIC ACID; TYROSINE PHOSPHORYLATION; SERUM-ALBUMIN; MOTILITY; PROTEIN; MATRIX; TRANSDUCTION; METASTASIS; P21(RAS); ADHESION AB We have pioneered an in vitro pseudopod-generation model wherein suspended tumor cells are stimulated to form pseudopods into glass micropipettes in response to soluble collagen type IV (CIV), Pertussis toxin and removing intracellular calcium were found previously to be inhibitory to that process. We now extend those observations to dissect the roles of transmembrane calcium influx and circulating fatty acids on pseudopod extension, Removal of fatty acids from BSA in basal media resulted in abrogation of pseudopod formation, while reconstitution of free fatty acids restored cell pseudopod protrusion. We thus hypothesized that fatty acids may provide necessary pseudopod stimulatory signals, Addition of lysophosphatidic acid (LPA) to the fatty acid-free CIV solution or in an opposite pipette without CIV permitted approximately 50% pseudopod recovery in all pipette directions in a dose-dependent fashion. Thapsigargin (TC), an agent that releases internal calcium stores and causes opening of store-operated calcium channels, restored pseudopod protrusion up to 80% in CIV with fatty acid-free albumin, [Ca2+](i) release was non-additive when cells were stimulated by TG and LPA, suggesting overlapping [Ca2+](i) stores. The combination of TG and LPA in fatty acid-free albumin fully restored the pseudopod response to CIV, Addition of EGTA to chelate stimulatory media calcium blocked the pseudopod response to CIV in the presence of fatty acids. This indicates that pseudopod protrusion requires transmembrane calcium entry. Thus, extracellular lipids and calcium mobilization are required to complement CIV in pseudopod protrusion from suspended cells. (C) 2000 Wiley-Liss, Inc. C1 Penn State Univ, Dept Bioengn, University Pk, PA 16802 USA. NCI, Pathol Lab, Bethesda, MD 20892 USA. RP Dong, C (reprint author), Penn State Univ, Dept Bioengn, 229 Hallowell Bldg, University Pk, PA 16802 USA. FU NCI NIH HHS [R29 CA076434-03, R29 CA076434-04, R29 CA076434, R29 CA076434-02, CA76434] NR 34 TC 4 Z9 4 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 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD NOV 15 PY 2000 VL 88 IS 4 BP 593 EP 600 DI 10.1002/1097-0215(20001115)88:4<593::AID-IJC12>3.3.CO;2-F PG 8 WC Oncology SC Oncology GA 368TW UT WOS:000090134500012 PM 11058876 ER PT J AU Emanuel, EJ Fairclough, DL Emanuel, LL AF Emanuel, EJ Fairclough, DL Emanuel, LL TI Attitudes and desires related to euthanasia and physician-assisted suicide among terminally ill patients and their caregivers SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID ONCOLOGY PATIENTS; CARE GIVERS; FAMILIES; HEALTH; BURDEN; DEATH AB Context Euthanasia and physician-assisted suicide (PAS) are highly controversial is' sues. While there are studies of seriously ill patients' interest in euthanasia and PAS, there are no data on the attitudes and desires of terminally ill patients regarding these issues. Objective To determine the attitudes of terminally ill patients toward euthanasia and PAS, whether they seriously were considering euthanasia and PAS for themselves, the stability of their desires, factors associated with their desires, and the proportion of patients who die from these interventions. Design Prospective cohort of terminally ill patients and their primary caregivers surveyed twice between March 1996 and July 1997. Setting Outpatient settings in 5 randomly selected metropolitan statistical areas and 1 rural county. Participants A total of 988 patients identified by their physicians to be terminally ill with any disease except for human immunodeficiency virus infection (response ratel 87.4%) and 893 patient-designated primary caregivers (response rate, 97.6%). Main Outcome Measures Support for euthanasia or PAS in standard scenarios; patient-expressed considerations and discussions of their desire for euthanasia or PAS; hoarding of drugs for suicide; patient death by euthanasia or PAS; and patient-reported sociodemographic factors and symptoms related to these outcomes. Results Of the 988 terminally ill patients, a total of 60.2% supported euthanasia or PAS in a hypothetical situation, but only 10.6% reported seriously considering euthanasia or PAS for themselves. Factors associated with being less likely to consider euthanasia or PAS were feeling appreciated (odds ratio [OR], 0.65; 95% confidence interval [CI], 0.52-0.82), being aged 65 years or older (OR, 0.52; 95% CI, 0.34-0.82), and being African American (OR, 0.39; 95 % CI, 0.18-0.84). Factors associated with being more likely to consider euthanasia or PAS were depressive symptoms (OR, 1.25; 95 % Clr 1.05-1.49), substantial caregiving needs (OR, 1.09; 95 % CI, 1.01 - 1.17), and pain (OR, 1.26; 95% CI, 1.02-1.56). At the follow-up interview, half of the terminally ill patients who had considered euthanasia or PAS for themselves changed their minds, while an almost equal number began considering these interventions. Patients with depressive symptoms (OR, 5.29; 95% CI, 1.21-23.2) and dyspnea (OR, 1.68; 95% CI, 1.26-2.22) were more likely to change their minds to consider euthanasia or PAS. According to the caregivers of the 256 decedents, 14 patients (5.6%) had discussed asking the physician for euthanasia or PAS and 6 (2.5%) had hoarded drugs. Ultimately, of the 256 decedents 1 (0.4%) died by euthanasia or PAS, 1 unsuccessfully attempted suicide, and 1 repeatedly requested for her life to be ended but the family and physicians refused. Conclusions In this survey, a small proportion of terminally ill patients seriously considered euthanasia or PAS for themselves. Over a few months, half the patients changed their minds. Patients with depressive symptoms were more likely to change their minds;about desiring euthanasia or PAS. C1 NIH, Dept Clin Bioeth, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. Amer Med Ctr, Canc Res Ctr, Ctr Res Methodol & Biometr, Denver, CO USA. Northwestern Univ, Sch Med, Interdisciplinary Program Professionalism & Human, Chicago, IL USA. RP Emanuel, EJ (reprint author), NIH, Dept Clin Bioeth, Warren G Magnuson Clin Ctr, Bldg 10,Room 1C118, Bethesda, MD 20892 USA. NR 38 TC 187 Z9 188 U1 5 U2 28 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 NOV 15 PY 2000 VL 284 IS 19 BP 2460 EP 2468 DI 10.1001/jama.284.19.2460 PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 371MY UT WOS:000165183800019 PM 11074775 ER PT J AU Wolfe, J Klar, N Grier, HE Duncan, J Salem-Schatz, S Emanuel, EJ Weeks, JC AF Wolfe, J Klar, N Grier, HE Duncan, J Salem-Schatz, S Emanuel, EJ Weeks, JC TI Understanding of prognosis among parents of children who died of cancer - Impact on treatment goals and integration of palliative care SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID PEDIATRIC ONCOLOGY; PERSPECTIVES; PREFERENCES; GUIDELINES; PATIENT AB Context Parents' understanding of prognosis or decision making about palliative care for children who die of cancer is largely unknown. However, a more accurate under standing of prognosis could alter treatment goals and expectations and lead to more effective care. Objectives To evaluate parental understanding of prognosis in children who die of cancer and to assess the association of this factor with treatment goals and the palliative care received by children. Design, Setting, and Participants Survey, conducted between September 1997 and August 1998, of 103 parents of children who received treatment at the Dana-Farber Cancer Institute and Children's Hospital, Boston, Mass, and who died of cancer between 1990 and 1997 (72 % of those eligible and those located) and 42 pediatric oncologists. Main Outcome Measure Timing of parental understanding that the child had no realistic chance for cure compared with the timing of physician understanding of this prognosis, as documented in the medical record. Results Parents first recognized that the child had no realistic chance for cure a mean (SD) of 106 (150) days before the child's death, while physician recognition occurred earlier at 206 (330) days before death. Among children who died of progressive disease, the group characterized by earlier recognition of this prognosis by both parents and physicians had earlier discussions of hospice care (odds ratio [OR], 1.03; 95% confidence interval [CI], 1.01 -1.06; P=.01), better parental ratings of the quality of home care (OR, 3.31; 95% CI, 1.15-9.54; P=.03), earlier institution of a do-not-resuscitate order (OR, 1.03; 95% CI, 1.00-1.06; P=.02), less use of cancer-directed therapy during the last month of life (OR, 2.80; 95% CI, 1.05-7.50; P=.04), and higher likelihood that the goal of cancer-directed therapy identified by both physician and parent was to lessen suffering (OR, 5.17; 95% CI, 1.86-14.4; P=.002 for physician and OR, 6.56; 95% CI, 1.54-27.86; P=.01 for parents). Conclusion Considerable delay exists in parental recognition that children have no realistic chance for cure, but earlier recognition of this prognosis by both physicians and parents is associated with a stronger emphasis on treatment directed at lessening suffering and greater integration of palliative care. C1 Dana Farber Canc Inst, Ctr Outcomes & Policy Res, Boston, MA 02115 USA. Dana Farber Canc Inst, Dept Pediat Oncol, Boston, MA 02115 USA. Dana Farber Canc Inst, Dept Biostat Sci, Boston, MA 02115 USA. Dana Farber Canc Inst, Dept Adult Oncol, Boston, MA 02115 USA. Childrens Hosp, Dept Med, Boston, MA 02115 USA. Brigham & Womens Hosp, Dept Med, Boston, MA 02115 USA. Div Prevent Oncol Canc Care Ontario, Toronto, ON, Canada. NIH, Dept Clin Bioeth, Bethesda, MD 20892 USA. RP Wolfe, J (reprint author), Dana Farber Canc Inst, Ctr Outcomes & Policy Res, 44 Binney St, Boston, MA 02115 USA. FU AHRQ HHS [5 T32 HS00063] NR 31 TC 218 Z9 218 U1 2 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 NOV 15 PY 2000 VL 284 IS 19 BP 2469 EP 2475 DI 10.1001/jama.284.19.2469 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 371MY UT WOS:000165183800020 PM 11074776 ER PT J AU Krag, D Ashikaga, T Abrams, J AF Krag, D Ashikaga, T Abrams, J TI Sentinel nodes: Clinical trial data needed SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Letter C1 Univ Vermont, Burlington, VT 05405 USA. NCI, Bethesda, MD 20892 USA. RP Krag, D (reprint author), Univ Vermont, Burlington, VT 05405 USA. NR 1 TC 2 Z9 2 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD NOV 15 PY 2000 VL 18 IS 22 BP 3873 EP 3873 PG 1 WC Oncology SC Oncology GA 374NY UT WOS:000165352300016 PM 11078501 ER PT J AU Chen, CL Singh, N Yull, FE Strayhorn, D Van Kaer, L Kerr, LD AF Chen, CL Singh, N Yull, FE Strayhorn, D Van Kaer, L Kerr, LD TI Lymphocytes lacking I kappa B-alpha develop normally, but have selective defects in proliferation and function SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTOONCOGENE BCL-3 ENCODES; GERMINAL CENTER FORMATION; ANTIBODY-SECRETING CELLS; IMMUNE-RESPONSES; CD40 LIGAND; C-REL; TRANSCRIPTION FACTORS; CD40-DEFICIENT MICE; ONCOPROTEIN BCL-3; TRANSGENIC MICE AB NF-kappaB has been implicated in the development, activation, and function of B and T lymphocytes. We have evaluated the in vivo effects of deletion of I kappaB-alpha, a major inhibitor of NF-kappaB, on lymphocyte development, proliferation, and function. To elucidate the long term role of I kappaB-alpha ill lymphocytes, fetal liver cells of 14.5-day-old I kappaB-alpha (-/-) or wild-type embryos were transplanted into irradiated recombinase-activating gene-2-deficient mice. Within 4 wk, the I kappaB-alpha (-/-) fetal li, er cells reconstitute mature E and T cell populations in the recipients comparable to those produced by wild-type fetal liver cells, However, the proliferative responses of I kappaB-alpha (-/-) B cells are enhanced, whereas those of I kappaB-alpha (-/-) T cells are reduced. The levels of IgG1, IgG2a, IgA, and IgE produced by I kappaB-alpha (-/-) B cells are elevated relative to those produced by I kappaB-alpha (+/+) Or I kappaB-alpha (+/-). Moreover, the specific immune responses to OVA and the generation of germinal centers are impaired in recipients of I kappaB-alpha (-/-) fetal liver cells, These results indicate that I kappaB-alpha plays a vital role in signal transduction pathways regulating lymphocyte proliferation and also in the production of specific Ig isotypes. C1 Vanderbilt Univ, Sch Med, Dept Microbiol & Immunol, Nashville, TN 37232 USA. Vanderbilt Univ, Sch Med, Dept Cell Biol, Nashville, TN 37232 USA. Howard Hughes Med Inst, Chevy Chase, MD 20815 USA. Vanderbilt Ingram Canc Ctr, Nashville, TN 37232 USA. NIH, Div Allergy Immunol & Infect Dis, Transplantat & Immunobiol Sect, Bethesda, MD 20892 USA. RP Yull, FE (reprint author), Vanderbilt Univ, Sch Med, Dept Microbiol & Immunol, MCN A4314,1161 21st Ave S, Nashville, TN 37232 USA. RI Van Kaer, Luc/H-1033-2015 OI Van Kaer, Luc/0000-0001-5275-2309 FU NCI NIH HHS [CA68485, P30CA68485]; NIGMS NIH HHS [R01GM51249] NR 45 TC 28 Z9 28 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 NOV 15 PY 2000 VL 165 IS 10 BP 5418 EP 5427 PG 10 WC Immunology SC Immunology GA 372VK UT WOS:000165254600007 PM 11067893 ER PT J AU Wahlsten, JL Gitchell, HL Chan, CC Wiggert, B Caspi, RR AF Wahlsten, JL Gitchell, HL Chan, CC Wiggert, B Caspi, RR TI Fas and Fas ligand expressed on cells of the immune system, not on the target tissue, control induction of experimental autoimmune uveitis SO JOURNAL OF IMMUNOLOGY LA English DT Article ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; BINDING PROTEIN IRBP; VS-HOST DISEASE; LYMPHOPROLIFERATIVE DISEASE; APOPTOSIS; MICE; UVEORETINITIS; ACTIVATION; GENE; LPR AB The Fas-Fas ligand (FasL) interaction is important for maintaining lymphocyte homeostasis by signaling for activation-induced cell death, Mice homozygous for the lpr or gld mutations do not express functional Fas or Fast, respectively; and spontaneously develop progressive autoimmune symptoms. Recent studies implicated expression of Fast on Immunologically privileged tissues in protection from immune-mediated damage. Conversely, tissue expression of Fas may facilitate damage. We evaluated the susceptibility of lpr and gld mice to induction of experimental autoimmune uveitis (EAU), a T cell-mediated autoimmune disease induced with retinal Ags, which targets the neural retina. gld as well as lpr mice immunized with a retinal Ag developed disease of lower incidence and severity than wild-type controls. Delayed hypersensitivity responses were not significantly different among immunized gld, lpr, or wild-type mice, although in vitro Ag-specific lymphocyte responses of the mutant mice were lower, To evaluate whether the diminished ability of gld and lpr mice to develop EAU was due to a defect at the level of the tissue or the immune system, radiation bone marrow chimeras constructed between wild-type and mutant mice were immunized to induce EAU. Mutant recipients of,wild-type bone marrow, but not wild-type recipients of mutant bone marrow, developed normal disease scores, These results indicate that normal expression of Fas and of Fast on cells of the immune system is important for EAU expression, Unexpectedly, neither lack of Fas nor lack of Fast on the ocular tissues affected expression of EAU. C1 NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. NEI, Retinal Cell & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Caspi, RR (reprint author), NEI, Immunol Lab, NIH, 10 Ctr Dr,10-10N222, Bethesda, MD 20892 USA. NR 34 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 NOV 15 PY 2000 VL 165 IS 10 BP 5480 EP 5486 PG 7 WC Immunology SC Immunology GA 372VK UT WOS:000165254600014 PM 11067900 ER PT J AU Polakova, K Plaksin, D Chung, DH Belyakov, IM Berzofsky, JA Margulies, DH AF Polakova, K Plaksin, D Chung, DH Belyakov, IM Berzofsky, JA Margulies, DH TI Antibodies directed against the MHC-I molecule H-2D(d) complexed with an antigenic peptide: Similarities to a T cell receptor with the same specificity SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MAJOR HISTOCOMPATIBILITY COMPLEX; SURFACE-PLASMON RESONANCE; MONOCLONAL-ANTIBODY; CRYSTAL-STRUCTURE; RESTRICTED SPECIFICITY; ANGSTROM RESOLUTION; LIGAND RECOGNITION; ENVELOPE PROTEIN; 2.5 ANGSTROM; SELF-PEPTIDE AB alpha beta TCRs, which use an Ab-like structure to form a combining site, recognize molecular complexes consisting of peptides bound to MHC class I(MHC-I) or class II (MHC-II) molecules, To explore the similarities and differences between Ab acid T cell recognition of similar structures, we have isolated two mAbs, KP14 and KP15, that specifically bind H-2D(d) complexed with an HIV envelope gp160-derived peptide, P18-I10. These Abs are MHC and peptide specific, Fine specificity of mAb binding was analyzed using a panel of synthetic peptides, revealing similarities between the mAb and a cloned TCR with the same specificity. These two mAbs used the same V-H and J(H) gene segments, but different D, V kappa, and J kappa genes. Administered in vivo, mAb KP15 blocked the induction of CTL specific for recombinant vaccinia virus-encoded gp160, indicating its ability to bind endogenously generated MHC/peptide complexes. Analysis of the fine specificity of these mAbs in the context of their encoded amino acid sequences and the known three-dimensional structure of the H-2D(d)/P18-I10 complex suggests that they bind in an orientation similar to that of the TCR, Thus, the plasticity of the B cell receptor repertoire and the structural similarities among BCR and TCR allow Abs to effectively mimic alpha beta TCRs. Such mAbs may be useful in the therapeutic modulation of immune responses against infectious agents or harmful self Ags as well as in tracing steps in Ag processing. C1 NIAID, Mol Biol Sect, Immunol Lab, NIH, Bethesda, MD 20892 USA. NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. RP Margulies, DH (reprint author), NIAID, Mol Biol Sect, Immunol Lab, NIH, Bldg 10,Room 11N311, Bethesda, MD 20892 USA. RI Chung, Doo Hyun/J-2791-2012; Margulies, David/H-7089-2013; OI Margulies, David/0000-0001-8530-7375 NR 79 TC 22 Z9 23 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 NOV 15 PY 2000 VL 165 IS 10 BP 5703 EP 5712 PG 10 WC Immunology SC Immunology GA 372VK UT WOS:000165254600042 PM 11067928 ER PT J AU Hoffmann, TK Nakano, K Elder, EM Dworacki, G Finkelstein, SD Appella, E Whiteside, TL DeLeo, AB AF Hoffmann, TK Nakano, K Elder, EM Dworacki, G Finkelstein, SD Appella, E Whiteside, TL DeLeo, AB TI Generation of T cells specific for the wild-type sequence p53(264-272) peptide in cancer patients: Implications for immunoselection of epitope loss variants SO JOURNAL OF IMMUNOLOGY LA English DT Article ID DENDRITIC CELLS; TARGETING P53; NECK-CANCER; LYMPHOCYTES; MUTATIONS; ANTIGEN; HEAD; IMMUNOTHERAPY; CYTOTOXICITY; CARCINOMAS AB Alterations in the p53 gene occur frequently and can lead to accumulation of p53 protein in squamous cell carcinomas of the head and neck (SCCHN), Since accumulation of p53 is associated with enhanced presentation of wild-type sequence (wt) p53 peptides to immune cells, the development of pan vaccines against SCCHN has focused on,vt p53 epitopes, We used the HLA-A2.1-restricted wt p53(264-272) epitope to generate CTL from circulating precursor T cells of HLA-A2.1(+) healthy donors and patients with SCCHN, Autologous peptide-pulsed dendritic cells were used for in vitro sensitization, CTL specific for the wt p53(264-272) peptide were generated from PBMC obtained from two of seven normal donors and three of seven patients with SCCHN, These CTL were HLA class I restricted and responded to T2 cells pulsed with p53(264-272) peptide as well as HLA-A2-matched SCCHN cell lines naturally presenting the epitope, Paradoxically, none of the tumors in the three patients who generated CTL could adequately present the epitope; two had a nt p53 genotype and no p53 protein accumulation, while the third tumor expressed a point mutation (R to H) in codon 273 that prevents presentation of the p53(264-272) epitope, In contrast, patients who did not generate CTL had tumors that accumulated altered p53 and potentially could present the p53(264-272) epitope. These findings suggest that in vivo, CTL specific for the wt p53(264-272) peptide might play a role in the elimination of tumor cells expressing this epitope and in immunoselection of epitope-loss tumor cells, Immunoselection of tumors that become resistant to anti-p53 immune responses has important implications for future p53-based vaccination strategies. C1 Univ Pittsburgh, Pittsburgh Canc Inst, Pittsburgh, PA 15213 USA. Univ Pittsburgh, Sch Med, Dept Pathol, Pittsburgh, PA 15213 USA. Univ Pittsburgh, Sch Med, Dept Otolaryngol, Pittsburgh, PA 15213 USA. NCI, Bethesda, MD 20892 USA. RP Whiteside, TL (reprint author), Univ Pittsburgh, Pittsburgh Canc Inst, W1041 Biomed Sci Tower,211 Lothrop St, Pittsburgh, PA 15213 USA. FU NIDCR NIH HHS [P01-DE12321] NR 40 TC 63 Z9 65 U1 0 U2 4 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD NOV 15 PY 2000 VL 165 IS 10 BP 5938 EP 5944 PG 7 WC Immunology SC Immunology GA 372VK UT WOS:000165254600070 PM 11067956 ER PT J AU Odendahl, M Jacobi, A Hansen, A Feist, E Hiepe, F Burmester, GR Lipsky, PE Radbruch, A Dorner, T AF Odendahl, M Jacobi, A Hansen, A Feist, E Hiepe, F Burmester, GR Lipsky, PE Radbruch, A Dorner, T TI Disturbed peripheral B lymphocyte homeostasis in systemic lupus erythematosus SO JOURNAL OF IMMUNOLOGY LA English DT Article ID V(H)4-21 GENE SEGMENT; DEPENDENT ANTIBODY-RESPONSES; T-CELL ACTIVATION; PLASMA-CELLS; BONE-MARROW; CD27/CD70 INTERACTION; SOMATIC MUTATION; SECRETING CELLS; I SPECIFICITY; BLOOD AB In patients with active systemic lupus erythematosus (SLE), a marked B lymphocytopenia was identified that affected CD19(+)/CD27(-) naive B cells more than CD19(+)/CD27(+) memory B cells, leading to a relative predominance of CD27-expressing peripheral B cells. CD27(high)/CD38(+)/CD19(dim)/surface Ig(low)/CD20(-)/CD138(+) plasma cells were found at high frequencies In active but not inactive SLE patients. Upon immunosuppressive therapy, CD27(high) plasma cells and naive CD27(-) B cells were markedly decreased in the peripheral blood, Mutational analysis of V gene rearrangements of individual B cells confirmed that CD27(+) B cells coexpressing IgD were memory B cells preferentially using V(H)3 family members with multiple somatic mutations. CD27(high) plasma cells showed a similar degree of somatic hypermutation, but preferentially employed V(H)4 family members, These results indicate that there are profound abnormalities in the various B cell compartments in SLE that respond differently to immunosuppressive therapy. C1 Univ Hosp Charite, Dept Med Rheumatol & Clin Immunol, D-10098 Berlin, Germany. Univ Hosp Charite, Outpatients Dept, D-10098 Berlin, Germany. Deutsch Rheuma Forschungszentrum, Berlin, Germany. NIAMSD, NIH, Bethesda, MD 20892 USA. RP Dorner, T (reprint author), Univ Hosp Charite, Dept Med Rheumatol & Clin Immunol, Schumannstr 20-21, D-10098 Berlin, Germany. RI Hiepe, Falk/A-9416-2013 NR 54 TC 346 Z9 357 U1 1 U2 16 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD NOV 15 PY 2000 VL 165 IS 10 BP 5970 EP 5979 PG 10 WC Immunology SC Immunology GA 372VK UT WOS:000165254600074 PM 11067960 ER PT J AU Tschetter, JR Mozes, E Shearer, GM AF Tschetter, JR Mozes, E Shearer, GM TI Progression from acute to chronic disease in a murine parent-into-F-1 model of graft-versus-host disease SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TOXIC LYMPHOCYTES-T; CELLS; LUPUS; MICE; EXPRESSION; ACTIVATION; ANTIBODIES; AUTOANTIBODIES; REPOPULATION; INDUCTION AB The parent-into-immunocompetent-F-1 model of graft-vs-host disease (GVHD) induces immune dysregulation, resulting in acute or chronic GVHD. The disease outcome is thought to be determined by the number of parental anti-F-1 CTL precursor cells present in the inoculum. Injection of C57BL/6 (B6) splenocytes into (B6 x DBA/2)F-1 (B6D2F(1)) mice (acute model) leads to extensive parental cell engraftment and early death, whereas injection of DBA/2 cells (chronic model) results in little parental cell engraftment and a lupus-like disease. This study demonstrated that injection of BALB/c splenocytes into (BALB/c x B6)F-1 (CB6F(1)) mice resulted in little engraftment of parental lymphocytes and the development of lupus as expected. Injection of B6 splenocytes into CB6F(1) initiated an initial burst of parental cell engraftment similar to that of B6 into B6D2F(1). However, the acute disease resolved, and the CB6F(1) mice went on to develop chronic GVHD with detectable Abs to ssDNA, dsDNA, and extractable nuclear Ags. Limiting dilution CTL assays determined that B6 splenocytes have CTL precursor frequencies of 1/1000 against both CB6F(1) and B6D2F(1), whereas DBA/2 and BALB/c splenocytes have a CTL precursor frequency of 1/20,000 for their respective F(1)s. The Th cell precursor frequency for B6 anti-DBA/2 was 3-fold higher than that for B6 anti-BALB/c determined by limiting dilution proliferation assays. These results indicate the importance of adequate allospecific helper as well as effector T cells for the induction and maintenance of acute GVHD in this model, and presents an unexpected model in which initial acute GVHD is replaced by the chronic form of disease. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. Weizmann Inst Sci, Dept Immunol, IL-76100 Rehovot, Israel. RP Shearer, GM (reprint author), NCI, Expt Immunol Branch, NIH, 10 Ctr Dr,Bldg 10,Room 4B36, Bethesda, MD 20892 USA. OI Tschetter, Jolynne/0000-0003-1743-1221 NR 30 TC 38 Z9 42 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 NOV 15 PY 2000 VL 165 IS 10 BP 5987 EP 5994 PG 8 WC Immunology SC Immunology GA 372VK UT WOS:000165254600076 PM 11067962 ER PT J AU Reshkin, SJ Guerra, L Bagorda, A Debellis, L Cardone, R Li, AH Jacobson, KA Casavola, V AF Reshkin, SJ Guerra, L Bagorda, A Debellis, L Cardone, R Li, AH Jacobson, KA Casavola, V TI Activation of A(3) adenosine receptor induces calcium entry and chloride secretion in A(6) cells SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE A(3) receptor; C1-IB-MECA; A(6) cells; renal; chloride; calcium ID CORTICAL COLLECTING DUCT; MOLECULAR-CLONING; G-PROTEIN; A6 CELLS; EPITHELIAL-CELLS; SODIUM-TRANSPORT; CL CHANNELS; APICAL CL; LINE; VASOPRESSIN AB We have previously demonstrated that in A(6) renal epithelial cells, a commonly used model of the mammalian distal section of the nephron, adenosine A(1) and A(2A) receptor activation modulates sodium and chloride transport and intracellular pH (Casavola et al., 1997). Here we show that apical addition of the A(3) receptor-selective agonist, 2-chloro-N-6-(3-iodobenzyl)-adenosine-5' -methyluronamide (Cl-IB-MECA) stimulated a chloride secretion that was mediated by calcium- and cAMP-regulated channels. Moreover, in single cell measurements using the fluorescent dye Fura 2-AM, Cl-IB-MECA caused an increase in Ca2+ influx. The agonist-induced rise in [Ca2+](i), was significantly inhibited by the selective adenosine A(3) receptor antagonists, 2,3-diethyl-4,5-dipropyl-6-phenylpyridine-3-thio-carboxylate-5-carboxylate (MRS 1523) and 3-ethyl 5-benzyl 2-methyl-6-phenyl-4-phenylethynyl-1,4-(+/-)dihydropyridine-3,5-dicarboxylate (MRS 1191) but not by antagonists of either A(1) or A(2) receptors supporting the hypothesis that Cl-IB-MECA increases [Ca2+](i) by interacting exclusively with A(3) receptors. Cl-IB-MECA-elicited Ca2+ entry was not significantly inhibited by per tussis toxin pretreatment while being stimulated by cholera toxin preincubation or by raising cellular cAMP levels with forskolin or rolipram. Preincubation with the protein kinase A inhibitor, H89, blunted the Cl-IB-MECA-elicited [Ca2+](i) response. Moreover, Cl-IB-MECA elicited an increase in cAMP production that was inhibited only by an A(3) receptor antagonist. Altogether, these data suggest that in A(6) cells a G(s)/protein kinase A pathway is involved in the A(3) receptor-dependent increase in calcium entry. C1 Univ Bari, Dept Gen & Environm Physiol, I-70126 Bari, Italy. NIDDK, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Casavola, V (reprint author), Univ Bari, Dept Gen & Environm Physiol, I-70126 Bari, Italy. RI Jacobson, Kenneth/A-1530-2009; OI Jacobson, Kenneth/0000-0001-8104-1493; Debellis, Lucantonio/0000-0002-7571-0969 NR 51 TC 27 Z9 27 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 NOV 15 PY 2000 VL 178 IS 2 BP 103 EP 113 DI 10.1007/s002320010018 PG 11 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 373YV UT WOS:000165318400003 PM 11083899 ER PT J AU Clay, JR Kuzirian, AM AF Clay, JR Kuzirian, AM TI Localization of voltage-gated K+ channels in squid giant axons SO JOURNAL OF NEUROBIOLOGY LA English DT Article DE axoplasmic transport; ion channel; kinesin; protein recycling ID SYNAPTIC VESICLE; NA+ CHANNELS; TRANSPORT; MEMBRANE; NEURONS; LOLIGO; IDENTIFICATION; AXOPLASM; FUSION AB We have localized the classical voltage-gated K+ channel within squid giant axons by immunocytochemistry using the Kv1 antibody of Rosenthal et al, (1996), Widely dispersed patches of intense immunofluorescence were observed in the axonal membrane. Punctate immunofluorescence was also observed in the axoplasm and was localized to similar to 25-50-mum-wide column down the length of the nerve (axon diameter similar to 500 mum). Immunoelectronmicroscopy of the axoplasm revealed a K+ channel containing vesicles, 30-50 nm in diameter, within this column, These and other vesicles of similar size were isolated from axoplasm using a novel combination of high-speed ultracentrifugation and controlled-pore size, glass bead separation column techniques. Approximately 1% of all isolated vesicles were labeled by K+ channel immunogold reacted antibody. Incorporation of isolated vesicle fractions within an artificial lipid bilayer revealed K+ channel electrical activity similar to that recorded directly from the axonal membrane by Llano ct al, (1988), These K+ channel-containing vesicles may be involved in cycling of K+ channel protein into the axonal membrane, We have also isolated an axoplasmic fraction containing similar to 150-nm-diameter vesicles that may transport K+ channels back to the cell body. (C) 2000 John Wiley & Sons, Inc. C1 NINDS, Neurophysiol Lab, NIH, Bethesda, MD 20892 USA. Marine Biol Lab, Woods Hole, MA 02543 USA. RP NINDS, Neurophysiol Lab, NIH, Bethesda, MD 20892 USA. EM jrclay@ninds.nih.gov NR 35 TC 11 Z9 11 U1 1 U2 2 PU JOHN WILEY & SONS INC PI HOBOKEN PA 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0022-3034 J9 J NEUROBIOL JI J. Neurobiol. PD NOV 15 PY 2000 VL 45 IS 3 BP 172 EP 184 DI 10.1002/1097-4695(20001115)45:3<172::AID-NEU5>3.0.CO;2-T PG 13 WC Neurosciences SC Neurosciences & Neurology GA 374MM UT WOS:000165349000005 PM 11074463 ER PT J AU Toth, K Suares, G Lawrence, JJ Tansey, EP McBain, CJ AF Toth, K Suares, G Lawrence, JJ Tansey, EP McBain, CJ TI Differential mechanisms of transmission at three types of mossy fiber synapse SO JOURNAL OF NEUROSCIENCE LA English DT Article DE interneuron; GABAergic; Ca-permeable; AMPA receptor; LTP; mGluR ID GLUTAMATE-RECEPTOR CHANNELS; LONG-TERM POTENTIATION; NEOCORTICAL PYRAMIDAL NEURONS; PAIRED-PULSE FACILITATION; RAT HIPPOCAMPUS; AMPA RECEPTORS; CA1 REGION; INHIBITORY NEURONS; DENTATE GYRUS; PLASTICITY AB The axons of the dentate gyrus granule cells, the so-called mossy fibers, innervate their inhibitory interneuron and pyramidal neuron targets via both anatomically and functionally specialized synapses. Mossy fiber synapses onto inhibitory interneurons were comprised of either calcium-permeable (CP) or calcium-impermeable (CI) AMPA receptors, whereas only calcium-impermeable AMPA receptors existed at CA3 principal neuron synapses. In response to brief trains of high-frequency stimuli (20 Hz), pyramidal neuron synapses invariably demonstrated shortterm facilitation, whereas interneuron EPSCs demonstrated either short-term facilitation or depression. Facilitation at all CI AMPA synapses was voltage independent, whereas EPSCs at CP AMPA synapses showed greater facilitation at -20 than at -80 mV, consistent with a role for the postsynaptic unblock of polyamines. At pyramidal cell synapses, mossy fiber EPSCs possessed marked frequency-dependent facilitation (commencing at stimulation frequencies >0.1 Hz), whereas EPSCs at either type of interneuron synapse showed only moderate frequency-dependent facilitation or underwent depression. Presynaptic metabotropic glutamate receptors (mGluRs) decreased transmission at all three synapse types in a frequency-dependent manner. However, after block of presynaptic mGluRs, transmission at interneuron synapses still did not match the dynamic range of EPSCs at pyramidal neuron synapses. High-frequency stimulation of mossy fibers induced long-term potentiation (LTP), long-term depression (LTD), or no change at pyramidal neuron synapses, interneuron CP AMPA synapses, and CI AMPA synapses, respectively. Induction of LTP or LTD altered the shortterm plasticity of transmission onto both pyramidal cells and interneuron CP AMPA synapses by a mechanism consistent with changes in release probability. These data reveal differential mechanisms of transmission at three classes of mossy fiber synapse made onto distinct targets. C1 NICHHD, Lab Cellular & Mol Neurophysiol, NIH, Bethesda, MD 20892 USA. RP McBain, CJ (reprint author), Room 5A72,Bldg 49,Convent Dr, Bethesda, MD 20892 USA. RI yu, yan/C-2322-2012 NR 54 TC 153 Z9 158 U1 0 U2 13 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 NOV 15 PY 2000 VL 20 IS 22 BP 8279 EP 8289 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 370PM UT WOS:000165131500012 PM 11069934 ER PT J AU Hertz, L Chen, Y Spatz, M AF Hertz, L Chen, Y Spatz, M TI Involvement of non-neuronal brain cells in AVP-mediated regulation of water space at the cellular, organ, and whole-body level SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Review DE aquaporin; astrocytes; brain endothelial cells; choroid plexus; hypothalamic astrocytes; microvessels; pituicytes ID RAT CHOROID-PLEXUS; MAGNOCELLULAR NEUROSECRETORY-SYSTEM; MICROVASCULAR ENDOTHELIAL-CELLS; GLIAL LIMITANS SUBJACENT; CENTRAL-NERVOUS-SYSTEM; ARGININE-VASOPRESSIN; CEREBROSPINAL-FLUID; SUPRAOPTIC NUCLEUS; EXTRACELLULAR POTASSIUM; VOLUME REGULATION AB Vasopressin (AVP) influences non-neuronal brain cells in cell-type specific manners: (1) it regulates water balance at the cellular level of brain parenchyma by adjusting astrocytic water permeability; (2) it contributes to the control of extracellular K(+) concentration ([K(+)](e)) in brain by stimulation of K(+) transfer from blood to brain, due to activation of an inwardly directed Na(+),K(+),Cl(-) cotransporter at the luminal membrane of capillary endothelial cells and opening of K+ channels at their abluminal membrane; (3) it decreases formation of cerebrospinal fluid (CSF) by decreasing Cl(-) secretion into CSF by epithelial cells of the choroid plexus, probably by inhibition of Cl(-)/HCO(3)(-) exchange at their basolateral membrane; (4) it contributes to regulation of intracellular volume within the brain by regulation of water permeability in ependymal cells and subpial astrocytes; and (5) it exerts effects on specialized astrocytes in circumventricular organs, their adjacent glia limitans, and the neural pituitary, which regulate AVP release to the systemic circulation by altering the spatial relationship between neurons and their adjacent glial cells. A unified mechanism is proposed, which integrates most of the effects of AVP and may be of considerable importance for neuronal excitability and, thus, for behavior. (C) 2000 Wiley-Liss, Inc. C1 NINDS, Stroke Branch, NIH, Bethesda, MD 20892 USA. Univ Saskatchewan, Dept Pharmacol, Saskatoon, SK S7N 0W0, Canada. RP Hertz, L (reprint author), RR2,Box 245, Gilmour, ON K0L 1W0, Canada. EM lhertz@northcom.net NR 127 TC 21 Z9 22 U1 0 U2 0 PU WILEY-BLACKWELL PI MALDEN PA COMMERCE PLACE, 350 MAIN ST, MALDEN 02148, MA USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD NOV 15 PY 2000 VL 62 IS 4 BP 480 EP 490 DI 10.1002/1097-4547(20001115)62:4<480::AID-JNR2>3.0.CO;2-K PG 11 WC Neurosciences SC Neurosciences & Neurology GA 369YE UT WOS:000165094700002 PM 11070491 ER PT J AU Kepplinger, KJF Forstner, G Kahr, H Leitner, K Pammer, P Groschner, K Soldatov, NM Romanin, C AF Kepplinger, KJF Forstner, G Kahr, H Leitner, K Pammer, P Groschner, K Soldatov, NM Romanin, C TI Molecular determinant for run-down of L-type Ca2+ channels localized in the carboxyl terminus of the alpha(1C) subunit SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Article ID VOLTAGE-DEPENDENT INACTIVATION; GUINEA-PIG MYOCYTES; CALCIUM CHANNELS; MEMBRANE DEPOLARIZATION; CREB PHOSPHORYLATION; HIPPOCAMPAL-NEURONS; CYTOPLASMIC FACTOR; OPEN PROBABILITY; PROTEIN-KINASE; CALPASTATIN AB 1. The role of the sequence 1572-1651 in the C-terminal tail of the alpha (1C) subunit in run-down of Ca2+ channels was studied by comparing functional properties of the conventional alpha (1C,77) channel with those of three isoforms carrying alterations in this motif. 2. The pore-forming alpha (1C) subunits were co-expressed with alpha (2)delta and beta (2a) subunits in HEK-tsA201 cells, a subclone of the human embryonic kidney cell line, and studied by whole-cell and single-channel patch-clamp techniques. 3. Replacement of amino acids 1572-1651 in alpha (1C,77) with 81 different amino acids leading to alpha (1C,86) significantly altered run-down behaviour. Run-down of Ba2+ currents was rapid with alpha (1C,77) channels, but was slow with alpha (1C,86). 4. Transfer of the alpha (1C,86) segments L (amino acids 1572-1598) or K (amino acids 1595-1652) into the alpha (1C,77) channel yielded alpha (1C,77L) and alpha (1C,77K) channels, respectively, the run-down of which resembled more that of alpha (1C,77). These results demonstrate that a large stretch of sequence between residues 1572 and 1652 of alpha (1C,86) renders Ca2+ channels markedly resistant to run-down. 5. The protease inhibitor calpastatin added together with ATP was able to reverse the run-down of alpha (1C,77) channels. Calpastatin expression was demonstrated in the HEK-tsA cells by Western blot analysis. 6. These results indicate a significant role of the C-terminal sequence 1572-1651 of the alpha (1C) subunit in run-down of L-type Ca2+ channels and suggest this sequence as a target site for a modulatory effect by endogenous calpastatin. C1 Univ Linz, Inst Biophys, A-4040 Linz, Austria. Graz Univ, Inst Pharmacol & Toxicol, A-8010 Graz, Austria. NIA, NIH, Baltimore, MD 21224 USA. RP Romanin, C (reprint author), Univ Linz, Inst Biophys, Altenbergerstr 69, A-4040 Linz, Austria. RI Groschner, Klaus/A-2550-2010; Romanin, Christoph/D-5399-2009 OI Romanin, Christoph/0000-0003-3756-4136 NR 52 TC 21 Z9 23 U1 0 U2 2 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0022-3751 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD NOV 15 PY 2000 VL 529 IS 1 BP 119 EP 130 DI 10.1111/j.1469-7793.2000.00119.x PG 12 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 378MZ UT WOS:000165589300013 PM 11080256 ER PT J AU Zanger, D Yang, BK Ardans, J Waclawiw, MA Csako, G Wahl, LM Cannon, RO AF Zanger, D Yang, BK Ardans, J Waclawiw, MA Csako, G Wahl, LM Cannon, RO TI Divergent effects of hormone therapy on serum markers of inflammation in postmenopausal women with coronary artery disease on appropriate medical management SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID C-REACTIVE PROTEIN; INTERCELLULAR-ADHESION MOLECULE-1; SUDDEN ISCHEMIC DEATH; SMOOTH-MUSCLE CELLS; CARDIOVASCULAR-DISEASE; MYOCARDIAL-INFARCTION; EXTRACELLULAR-MATRIX; REPLACEMENT THERAPY; HEART-DISEASE; E-SELECTIN AB OBJECTIVES The goal of our study was to determine whether hormone therapy alters markers of inflammation in postmenopausal women With chronic stable coronary artery disease (CAD) on appropriate medical management. BACKGROUND Hormone therapy reduces some markers of inflammation associated with atherosclerosis risk (cell adhesion molecules) but increases levels of another marker of inflammation-C-reactive protein-in healthy postmenopausal women. METHODS Ten women (average age 66 years; range 59 to 76 years) with CAD on medical management (including aspirin [9], statin lipid-lowering therapy [7], angiotensin-converting enzyme inhibitors [3]) were randomly assigned to conjugated equine estrogens 0.625 mg (combined with medroxyprogesterone acetate 2.5 mg daily in five women with uterus intact) or placebo(s) daily for one month with crossover to the alternate therapy after one month off of hormone treatment in a double-blind study. At the end of each treatment phase, the following markers of inflammation were measured in serum: interleukin-6, C-reactive protein, E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1 and matrix metalloproteinase-9. RESULTS Hormone therapy significantly lowered serum levels of cell adhesion molecules E-selectin (46.9 +/- 18.3 vs. 56.3 +/- 20.6 ng/mL, p = 0.006), intercellular adhesion molecule-1 (282 +/- 74 vs. 303 +/- 78 ng/mL, p = 0.013) and vascular cell adhesion molecule-1 (605 +/-: 218 vs. 657 +/- 214 ng/mL, p = 0.01) but increased levels of matrix metalloproteinase-9 (648 +/- 349 vs. 501 +/- 285 ng/mL, p = 0.02). Interleukin-6 (4.33 +/- 4.78 vs. 3.04 +/- 1.47 pg/mL, p = 0.283) and C-reactive protein (0.88 +/- 1.13 vs. 0.61 +/- 0.50 mg/dL, p = 0.358) were not significantly elevated on hormone therapy compared with placebo values. CONCLUSIONS Hormone therapy has divergent effects on serum markers of inflammation in women with CAD. Reduction in levels of cell adhesion molecules may reduce attachment of white blood cells to the vessel wall, but increases in matrix metalloproteinase-9 within the vessel wall could digest and weaken fibrous caps of vulnerable plaques, thus provoking thrombosis. (C) 2000 by the American College of Cardiology. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Immunopathol Sect, NIH, Bethesda, MD 20892 USA. NIH, Ctr Clin, Dept Clin Pathol, Bethesda, MD 20892 USA. RP Cannon, RO (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,Room 7B15,10 Ctr Dr MSC-1650, Bethesda, MD 20892 USA. NR 44 TC 136 Z9 136 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD NOV 15 PY 2000 VL 36 IS 6 BP 1797 EP 1802 DI 10.1016/S0735-1097(00)00952-9 PG 6 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 375FL UT WOS:000165388800008 PM 11092646 ER PT J AU Domanski, MJ Borkowf, CB Campeau, L Knatterud, GL White, C Hoogwerf, B Rosenberg, Y Geller, NL AF Domanski, MJ Borkowf, CB Campeau, L Knatterud, GL White, C Hoogwerf, B Rosenberg, Y Geller, NL CA Post CABG Trial Investigators TI Prognostic factors for atherosclerosis progression in saphenous vein grafts - The postcoronary artery bypass graft (post-CABG) trial SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID ANGIOGRAPHIC FOLLOW-UP; RISK-FACTORS; CORONARY-BYPASS; COLESTIPOL-NIACIN; SURGERY; DISEASE; SURVIVAL; REOPERATION; THERAPY; WOMEN AB OBJECTIVES The study was done to assess patients in the Post-Coronary Artery Bypass Graft (Post-CABG) trial to determine prognostic factors for atherosclerosis progression. BACKGROUND Saphenous vein grafts (SVGs) are effective in relieving angina and, in certain patient subsets, in prolonging life. However, the progression of atherosclerosis in many of these grafts limits their usefulness. METHODS The Post-CABG trial studied moderate versus aggressive lipid-lowering and low-dose warfarin versus placebo in patients with a history of coronary artery bypass surgery and found that more aggressive lipid lowering was effective in preventing progression of atherosclerosis in SVGs, but warfarin had no effect. Using variables measured at baseline, we sought the independent prognostic factors for atherosclerosis progression in SVGs, employing the statistical method of generalized estimating equations with a logit-link function. RESULTS Twelve independent prognostic factors for atherosclerosis progression were found. In the order of their importance they were: maximum stenosis of the graft at baseline angiography; years post-SVG placement; the moderate low-density lipoprotein- cholesterol (LDL-C) lowering strategy; prior myocardial infarction; high triglyceride level; small minimum graft diameter; low high-density lipoprotein-cholesterol (HDL-C); high LDL-C; high mean arterial pressure; low ejection fraction; male gender; and current smoking. CONCLUSIONS This study identified Post-CABG patient and SVG characteristics associated with saphenous vein graft; atherosclerosis progression. These data provide a basis for rational risk factor management to prevent progression of SVG atherosclerosis. (C) 2000 by the American College of Cardiology. C1 NHLBI, Clin Trials Grp, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, Bethesda, MD 20892 USA. Maryland Med Res Inst, Baltimore, MD USA. Univ Minnesota Hosp & Clin, Div Cardiol, Minneapolis, MN 55455 USA. Cleveland Clin Fdn, Cleveland, OH 44195 USA. Montreal Heart Inst, Dept Cardiol, Montreal, PQ H1T 1C8, Canada. RP Domanski, MJ (reprint author), NHLBI, Clin Trials Grp, Bldg 10, Bethesda, MD 20892 USA. NR 34 TC 67 Z9 70 U1 1 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD NOV 15 PY 2000 VL 36 IS 6 BP 1877 EP 1883 DI 10.1016/S0735-1097(00)00973-6 PG 7 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 375FL UT WOS:000165388800021 PM 11092659 ER PT J AU Feskanich, D Ziegler, RG Michaud, DS Giovannucci, EL Speizer, FE Willett, WC Colditz, GA AF Feskanich, D Ziegler, RG Michaud, DS Giovannucci, EL Speizer, FE Willett, WC Colditz, GA TI Prospective study of fruit and vegetable consumption and risk of lung cancer among men and women SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID BETA-CAROTENE; VITAMIN-C; UNITED-STATES; CARDIOVASCULAR-DISEASE; MALIGNANT NEOPLASMS; ALCOHOLIC BEVERAGE; NEW-JERSEY; EPIDEMIOLOGY; ANTIOXIDANTS; PREVENTION AB Background: Diets high in fruits and vegetables have been shown to be associated with a lower risk of lung cancer. beta -Carotene was hypothesized to be largely responsible for the apparent protective effect, but this hypothesis was not supported by clinical trials. Methods: We examined the association between lung cancer risk and fruit and vegetable consumption in 77283 women in the Nurses' Health Study and 47778 men in the Health Professionals' Follow-up Study. Diet was assessed with the use of a food-frequency questionnaire that included 15 fruits and 23 vegetables. We used logistic regression models to estimate relative risks (RRs) of lung cancer within each cohort, All statistical tests were two-sided, Results: We documented 519 lung cancer cases among the women and 274 among the men. Total fruit and vegetable consumption was associated with a modestly lower risk of lung cancer among the women but not among the men. The RR for the highest versus lowest quintile of intake was 0.79 (95% confidence interval [CI] = 0.59-1.06) among the women and 1.12 (95% CI = 0.74-1.69) among the men after adjustment for smoking status, quantity of cigarettes smoked per day, time since quitting smoking, and age at initiation of smoking. However, total fruit and vegetable consumption was associated with a lower risk of lung cancer among never smokers in the combined cohorts, although the reduction was not statistically significant (RR = 0.63; 95% CI = 0.35-1.12 in the highest tertile). Conclusion: Higher fruit and vegetable intakes were associated with lower risks of lung cancer in women but not in men. It is possible that the inverse association among the women remained confounded by unmeasured smoking characteristics, although fruits and vegetables were protective in both men and women who never smoked. C1 Brigham & Womens Hosp, Channing Lab, Dept Med, Boston, MA 02115 USA. Harvard Univ, Sch Med, Boston, MA USA. NCI, Epidemiol & Biostat Program, Div Canc Epidemiol & Genet, 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 Publ Hlth, Dept Environm Hlth, Boston, MA 02115 USA. Harvard Univ, Sch Publ Hlth, Ctr Canc Prevent, Boston, MA 02115 USA. RP Feskanich, D (reprint author), Brigham & Womens Hosp, Channing Lab, Dept Med, 181 Longwood Ave, Boston, MA 02115 USA. RI Michaud, Dominique/I-5231-2014; Colditz, Graham/A-3963-2009 OI Colditz, Graham/0000-0002-7307-0291 FU NCI NIH HHS [CA55075, CA40356] NR 51 TC 221 Z9 229 U1 2 U2 25 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD NOV 15 PY 2000 VL 92 IS 22 BP 1812 EP 1823 DI 10.1093/jnci/92.22.1812 PG 12 WC Oncology SC Oncology GA 372VH UT WOS:000165254400008 PM 11078758 ER PT J AU Koch, CA Eisenhofer, G Pacak, K AF Koch, CA Eisenhofer, G Pacak, K TI Diagnosing pheochromocytoma SO MEDIZINISCHE KLINIK LA German DT Article C1 NICHHD, Dept Hlth & Human Serv, Dev Endocrinol Branch, Bethesda, MD 20892 USA. RP Koch, CA (reprint author), NICHHD, Dept Hlth & Human Serv, Dev Endocrinol Branch, Bldg 10,Room 10N262,10 Ctr Dr,MSC 1862, Bethesda, MD 20892 USA. RI Koch, Christian/A-4699-2008; OI Koch, Christian/0000-0003-3127-5739; Koch, Christian/0000-0003-0678-1242 NR 4 TC 0 Z9 0 U1 0 U2 0 PU URBAN & VOGEL PI MUNICH PA NEUMARKTER STRASSE 43, D-81673 MUNICH, GERMANY SN 0723-5003 J9 MED KLIN JI Med. Klin. PD NOV 15 PY 2000 VL 95 IS 11 BP 655 EP 656 DI 10.1007/PL00002081 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 376CM UT WOS:000165440700011 PM 11143549 ER PT J AU Lemkin, PF Thornwall, GC Walton, KD Hennighausen, L AF Lemkin, PF Thornwall, GC Walton, KD Hennighausen, L TI The microarray explorer tool for data mining of cDNA microarrays: application for the mammary gland SO NUCLEIC ACIDS RESEARCH LA English DT Article ID ELECTROPHORETIC GEL IMAGES; 2DWG META-DATABASE; SACCHAROMYCES-CEREVISIAE; GENE-EXPRESSION; IDENTIFICATION; HYBRIDIZATION; PATTERNS; INTERNET AB The Microarray Explorer (MAExplorer) is a versatile Java-based data mining bioinformatic tool for analyzing quantitative cDNA expression profiles across multiple microarray platforms and DNA labeling systems. It may be run as either a stand-alone application or as a Web browser applet over the Internet, With this program it is possible to (i) analyze the expression of individual genes, (ii) analyze the expression of gene families and clusters, (iii) compare expression patterns and (iv) directly access other genomic databases for clones of interest. Data may be downloaded as required from a Web server or in the case of the stand-alone version, reside on the user's computer, Analyses are performed in real-time and maybe viewed and directly manipulated in images, reports,: scatter plots, histograms, expression profile plots and cluster analyses plots, A key feature is the clone data filter for constraining a working set of clones to those passing a variety of user-specified logical and statistical tests. Reports may be generated with hypertext Web access to UniGene, GenBank and other Internet databases for sets of clones found to be Of interest. Users may save their explorations on the Web Server or local computer and later recall or share them with other scientists in this groupware Web environment. The emphasis on direct manipulation of clones and sets of clones in graphics and tables provides a high level of interaction with the data, making it easier for investigators to test ideas when looking for patterns, We have used the MAExplorer to profile gene expression patterns of 1500 duplicated genes isolated from mouse mammary tissue. We have identified genes that are preferentially expressed during pregnancy and during lactation. One gene we identified, carbonic anhydrase III, is highly expressed in mammary tissue from virgin and pregnant mice and in. gene knock-out mice with underdeveloped mammary epithelium, Other genes, which include those encoding milk proteins, are preferentially expressed during lactation. MAExplorer may be accessed at http://www.lecb.ncifcrf.gov.MAExplorer. C1 NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, Frederick, MD 21702 USA. NIDDK, Lab Genet & Physiol, Bethesda, MD 20892 USA. FCRDC, SAIC, Frederick, MD 21702 USA. RP Lemkin, PF (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, Frederick, MD 21702 USA. EM lemkin@ncifcrf.gov OI Walton, Katherine/0000-0001-9108-5617 NR 31 TC 31 Z9 33 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD NOV 15 PY 2000 VL 28 IS 22 BP 4452 EP 4459 DI 10.1093/nar/28.22.4452 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 374VF UT WOS:000165364500007 PM 11071932 ER PT J AU He, LS Weber, A Levens, D AF He, LS Weber, A Levens, D TI Nuclear targeting determinants of the far upstream element binding protein, a c-myc transcription factor SO NUCLEIC ACIDS RESEARCH LA English DT Article ID HNRNP-K-PROTEIN; MESSENGER-RNA; LOCALIZATION SIGNAL; TERMINAL DOMAIN; DNA; SEQUENCE; IMPORT; MOTIF; IDENTIFICATION; VISUALIZATION AB FUSE binding protein (FBP) binds in vivo and in vitro with:the single-stranded far upstream element (FUSE) upstream of the c-myc gene, In addition to its transcriptional role, FBP and its closely related siblings FBP2 (KSRP) and FBP3 have been reported to bind-RNA and participate in various steps of RNA processing, transport or catabolism. To perform these diverse-functions, FBP must traffic to different nuclear-sites, To identify determinants of nuclear localization, full-length FBP or fragments thereof were fused to green fluorescent protein, Fluorescent-FBP-localized in the nucleus in three patterns, diffuse, dots and spots, Each pattern was conferred by a distinct nuclear localization signal (NLS): a classical bipartite: NLS in the N-terminal and two non-canonical signals, an alpha -helix in the third KH-motif of the nucleic acid binding domain and a tyrosine-rich motif in the C-terminal transcription activation domain, Upon treatment with the transcription inhibitor actinomycin D, FBP completely re-localized into dots, but did not exit from the nucleus, This is in contrast to many general RNA-binding proteins, which shuttle from the nucleus upon treatment with actinomycin D, Furthermore, FBP co-localized with transcription sites and with the general transcription factor TFIIH, but not:with the splicing factor SC-35, Taken together, these data reveal complex intranuclear trafficking of FBP and support a transcriptional role for this protein. C1 NCI, Pathol Lab, DCS, Bethesda, MD 20892 USA. RP Levens, D (reprint author), NCI, Pathol Lab, DCS, Bldg 10,Room 2N105, Bethesda, MD 20892 USA. EM levens@helix.nih.gov RI Levens, David/C-9216-2009 OI Levens, David/0000-0002-7616-922X NR 48 TC 39 Z9 41 U1 1 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 NOV 15 PY 2000 VL 28 IS 22 BP 4558 EP 4565 DI 10.1093/nar/28.22.4558 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 374VF UT WOS:000165364500021 PM 11071946 ER PT J AU Devedjiev, Y Dauter, Z Kuznetsov, SR Jones, TLZ Derewenda, ZS AF Devedjiev, Y Dauter, Z Kuznetsov, SR Jones, TLZ Derewenda, ZS TI Crystal structure of the human acyl protein thioesterase I from a single X-ray data set to 1.5 angstrom SO STRUCTURE LA English DT Article DE alpha/beta hydrolase; serine hydrolase; SAD; anomalous diffraction ID NEURONAL CEROID-LIPOFUSCINOSIS; ALPHA-SUBUNITS; LIPID MODIFICATIONS; PLASMA-MEMBRANE; PALMITOYLATION; ACTIVATION; RECEPTOR; LYSOPHOSPHOLIPASE; CRYSTALLOGRAPHY; PURIFICATION AB Background: Many proteins undergo posttranslational modifications involving covalent attachment of lipid groups. Among them is palmitoylation, a dynamic, reversible process that affects trimeric G proteins and Ras and constitutes a regulatory mechanism for signal transduction pathways. Recently, an acylhydrolase previously identified as lysophospholipase has been shown to function as an acyl protein thioesterase, which catalyzes depalmitoylation of Got proteins as well as Ras. Its amino acid sequence suggested that the protein is evolutionarily related to neutral lipases and other thioesterases, but direct structural information was not available. Results: We have solved the crystal structure of the human putative Ga-regulatory protein acyl thioesterase (hAPT1) with a single data set collected from a crystal containing the wildtype protein. The phases were calculated to 1.8 Angstrom resolution based on anomalous scattering from Br- ions introduced in the cryoprotectant solution in which the crystal was soaked for 20 s. The model was refined against data extending to a resolution of 1.5 Angstrom to an R factor of 18.6%. The enzyme is a member of the ubiquitous alpha/beta hydrolase family, which includes other acylhydrolases such as the palmitoyl protein thioesterase (PPT1). Conclusions: The human APT1 is closely related to a previously described carboxylesterase from Pseudomonas fluorescens. The active site contains a catalytic triad of Ser-114, His-203, and Asp-169. Like carboxylesterase, hAPT1 appears to be dimeric, although the mutual disposition of molecules in the two dimers differs. Unlike carboxylesterase, the substrate binding pocket and the active site of hAPT1 are occluded by the dimer interface, suggesting that the enzyme must dissociate upon interaction with substrate. C1 Univ Virginia, Hlth Sci Syst, Dept Mol Physiol & Biol Phys, Charlottesville, VA 22908 USA. Brookhaven Natl Lab, Upton, NY 11973 USA. NCI, Frederick, MD 21701 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Derewenda, ZS (reprint author), Univ Virginia, Hlth Sci Syst, Dept Mol Physiol & Biol Phys, Charlottesville, VA 22908 USA. FU NHLBI NIH HHS [HL48807] NR 49 TC 81 Z9 82 U1 1 U2 8 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTES AVE,, CAMBRIDGE, MA 02138 USA SN 0969-2126 J9 STRUCTURE JI Structure PD NOV 15 PY 2000 VL 8 IS 11 BP 1137 EP 1146 DI 10.1016/S0969-2126(00)00529-3 PG 10 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 420TX UT WOS:000168022300006 PM 11080636 ER PT J AU Wormser, U Sintov, A Brodsky, B Nyska, A AF Wormser, U Sintov, A Brodsky, B Nyska, A TI Topical iodine preparation as therapy against sulfur mustard-induced skin lesions SO TOXICOLOGY AND APPLIED PHARMACOLOGY LA English DT Article; Proceedings Paper CT 39th Annual Meeting of the American-Society-of-Toxicology CY MAR 19-24, 2000 CL PHILADELPHIA, PENNSYLVANIA SP Amer Soc Toxicol DE sulfur mustard; mustard gas; iodine; skin toxicity; dermatotoxicity ID INDUCED CUTANEOUS LESIONS; CHEMICAL WARFARE AGENT; HAIRLESS GUINEA-PIGS; PROTECTS A549 CELLS; ARGININE ANALOGS; IN-VITRO; APOPTOSIS; DNA; TOXICOLOGY; TOXICITY AB Sulfur mustard (SM) is a powerful vesicant employed as an agent of chemical warfare. This study demonstrates the therapeutic effect of a novel topical iodine preparation as a postexposure treatment against SM-induced lesions in the fur-covered guineapig skin model. Iodine treatment 15 min after SM exposure resulted in statistically significant reductions of 48, 50, and 55% in dermal acute inflammation, hemorrhage, and necrosis, respectively, whereas, the epidermal healing markers, hyperkerathosis and acanthosis, were significantly elevated by 72 and 67%, respectively, 2 days after treatment. At the interval of 30 min between SM exposure and iodine treatment, there was a significant degree of healing or recovery, albeit to a lesser extent than that observed in the shorter interval. Although the epidermal healing markers were not elevated, the parameters indicative of active tissue damage, such as subepidermal microblisters, epidermal ulceration, dermal. acute inflammation, hemorrhage, and necrosis, were significantly reduced by 35, 67, 43, 39, and 45%, respectively. At the 45-min interval between exposure and treatment, there was also a certain degree of healing or recovery expressed as significant reductions in dermal subacute inflammation, subepidermal microblister formation, and epidermal ulceration, whereas, acanthosis was statistically elevated, indicating an increased healing potential. At the 60-min interval, iodine was less efficacious; nevertheless, a significant reduction in the incidence of subepidermal microblisters and an expansion of the acanthotic area were observed. Gross ulceration was significantly decreased at intervals of 15 and 30 min between exposure and treatment. The local anesthetic, lidocaine, did not alter the therapeutic effect of iodine. SM was not affected chemically by iodine as measured by gas chromatography-mass spectrometry (GC-MS) analysis. These findings suggest that the iodine preparation functions as an antidote against skin lesions induced by SM. (C) 2000 Academic Press. C1 Hebrew Univ Jerusalem, Fac Sci, IL-91904 Jerusalem, Israel. Ben Gurion Univ Negev, Inst Appl Res, Beer Sheva, Israel. NIEHS, Lab Expt Pathol, NIH, Res Triangle Pk, NC 27709 USA. RP Wormser, U (reprint author), Hebrew Univ Jerusalem, Fac Sci, Berman Bldg,Edmond Safra Campus, IL-91904 Jerusalem, Israel. NR 35 TC 28 Z9 28 U1 0 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0041-008X J9 TOXICOL APPL PHARM JI Toxicol. Appl. Pharmacol. PD NOV 15 PY 2000 VL 169 IS 1 BP 33 EP 39 DI 10.1006/taap.2000.9056 PG 7 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 379EH UT WOS:000165627600005 PM 11076694 ER PT J AU Balbach, JJ Ishii, Y Antzutkin, ON Leapman, RD Rizzo, NW Dyda, F Reed, J Tycko, R AF Balbach, JJ Ishii, Y Antzutkin, ON Leapman, RD Rizzo, NW Dyda, F Reed, J Tycko, R TI Amyloid fibril formation by A beta(16-22), a seven-residue fragment of the Alzheimer's beta-amyloid peptide, and structural characterization by solid state NMR SO BIOCHEMISTRY LA English DT Article ID ANGLE-SPINNING NMR; MULTIPLE-QUANTUM NMR; C-13 CHEMICAL-SHIFT; MAGNETIC-RESONANCE SPECTROSCOPY; ATOMIC-FORCE MICROSCOPY; SYNCHROTRON X-RAY; MAGIC-ANGLE; ROTATIONAL-ECHO; EXCHANGE SPECTROSCOPY; SECONDARY STRUCTURE AB The seven-residue peptide N-acetyl-Lys-Leu-Val-Phe-Phe-Ala-Glu-NH2 called A beta (16-22) and representing residues 16-22 of the full-length beta -amyloid peptide associated with Alzheimer's disease, is shown by electron microscopy to form highly ordered fibrils upon incubation of aqueous solutions. X-ray powder diffraction and optical birefringence measurements confirm that these are amyloid fibrils, The peptide conformation and supramolecular organization in A beta (16-22) fibrils are investigated by solid state NMR measurements. Two-dimensional magic-angle spinning (2D MAS) exchange and constant-time double-quantum-filtered dipolar recoupling (CTDQFD) measurements indicate a beta -strand conformation of the peptide backbone at the central phenylalanine. One-dimensional and two-dimensional spectra of selectively and uniformly labeled samples exhibit C-13 NMR line widths of <2 ppm, demonstrating that the peptide, including amino acid side chains, has a well-ordered conformation in the fibrils. Two-dimensional C-13-C-13 chemical shift correlation spectroscopy permits a nearly complete assignment of backbone and side chain C-13 NMR signals and indicates that the -strand conformation extends across the entire hydrophobic segment from Leu17 through Ala21. C-13 multiple-quantum (MQ) NMR and C-13/N-15 rotational echo double-resonance (REDOR) measurements indicate an antiparallel organization of beta -sheets in the A beta (16-22), fibrils, These results suggest that the degree of structural order at the molecular level in amyloid fibrils can approach that in peptide or protein crystals, suggest how the supramolecular organization of beta -sheets in amyloid fibrils can be dependent on the peptide sequence, and illustrate the utility of solid state NMR measurements as probes of the molecular structure of amyloid fibrils, A beta (16-22) is among the shortest fibril-forming fragments of full-length beta -amyloid reported to date, and hence serves as a useful model system for physical studies of amyloid fibril formation. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. Lulea Univ Technol, Div Inorgan Chem, S-95187 Lulea, Sweden. NIH, Div Bioengn & Phys Sci, Off Res Serv, Bethesda, MD 20892 USA. NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Calif San Diego, La Jolla, CA 92093 USA. RP Tycko, R (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5,Room 112, Bethesda, MD 20892 USA. RI Reed, Jennifer/E-5137-2011; Ishii, Yoshitaka/F-4558-2014 OI Ishii, Yoshitaka/0000-0002-7724-6469 NR 82 TC 496 Z9 504 U1 8 U2 82 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD NOV 14 PY 2000 VL 39 IS 45 BP 13748 EP 13759 DI 10.1021/bi0011330 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375QM UT WOS:000165412800015 PM 11076514 ER PT J AU Misra, A Ozarowski, A Casas-Finet, JR Maki, AH AF Misra, A Ozarowski, A Casas-Finet, JR Maki, AH TI Effect of nucleic acid binding on the triplet state properties of tetrapeptides containing tryptophan and 6-methyltryptophan: A study by phosphorescence and ODMR spectroscopy SO BIOCHEMISTRY LA English DT Article ID DETECTED MAGNETIC-RESONANCE; HUMAN-IMMUNODEFICIENCY-VIRUS; MURINE LEUKEMIA-VIRUS; NUCLEOCAPSID PROTEIN; STACKING INTERACTIONS; AROMATIC RESIDUES; DNA; FLUORESCENCE; COMPLEXES; RNA AB Complexes of four peptides [KWGK, KGWK, K(6MeW)GK, KG(6MeW)K] with the nucleic acids [poly(A), poly(C), poly(U), poly(I), and rG(8)] have been investigated by phosphorescence and optically detected magnetic resonance (ODMR) spectroscopy. The intrinsic spectroscopic probes used in these studies are tryptophan (W) and 6-methyltryptophan (6MeW). Binding to the nucleic acids results in a red-shift of the phosphorescence 0,0-band (DeltaE(0,0)) of the aromatic residue as well as a reduction of its zero-field splitting parameter (DeltaD). Results are compared with earlier studies of the HIV-1 nucleocapsid protein, NCp7, that contains a single tryptophan residue (Trp37) within a retroviral zinc finger sequence. Binding of poly(A) or poly(U) to the tetrapeptides induces larger DeltaE(0,0) and DeltaD than when bound to NCp7, indicating stronger stacking interactions. Poly(I), on the ether hand, produces larger shifts in Trp37 of NCp7. Binding of rG(8) produces sequence-dependent effects in the peptides. When bound to NCp7, but in contrast with tetrapeptide binding, nucleic acids produce large changes in triplet state kinetics consistent with enhanced spin-orbit coupling. These results are discussed in terms of three limiting types of tryptophan-base interaction: intercalation, aromatic stacking, and edge-on interaction. These should have differing effects on the properties of the triplet state. C1 Univ Calif Davis, Dept Chem, Davis, CA 95616 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, AIDS Vaccine Program, Frederick, MD 21702 USA. RP Univ Calif Davis, Dept Chem, Davis, CA 95616 USA. EM maki@indigo.ucdavis.edu FU NIEHS NIH HHS [ES-02662]; PHS HHS [N01-56000] NR 59 TC 6 Z9 6 U1 1 U2 6 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD NOV 14 PY 2000 VL 39 IS 45 BP 13772 EP 13780 DI 10.1021/bi0015663 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375QM UT WOS:000165412800017 PM 11076516 ER PT J AU Zhu, JH Shearer, GM Norman, JE Pinto, LA Marincola, FM Prasad, A Waclawiw, MA Csako, G Quyyumi, AA Epstein, SE AF Zhu, JH Shearer, GM Norman, JE Pinto, LA Marincola, FM Prasad, A Waclawiw, MA Csako, G Quyyumi, AA Epstein, SE TI Host response to cytomegalovirus infection as a determinant of susceptibility to coronary artery disease - Sex-based differences in inflammation and type of immune response SO CIRCULATION LA English DT Article DE immunology; infection; inflammation; sex; risk factors ID ACUTE MYOCARDIAL-INFARCTION; HEART-DISEASE; CHLAMYDIA-PNEUMONIAE; VIRUS-INFECTION; RISK FACTOR; ATHEROSCLEROSIS; ASSOCIATION; TRANSPLANTATION; RESPONSIVENESS; HERPESVIRUS AB Background-Positive and negative associations between cytomegalovirus (CMV) infection and coronary artery disease (CAD) have been reported. We postulated that the susceptibility to CMV-induced CAD might relate to patterns of inflammatory and immune responses to CMV infection and that sex might have an effect on these responses. Methods and Results-In 151 men and 87 women being evaluated for CAD, blood samples were tested for humoral (Ab+) and cellular (Tc+) responses to CMV and for C-reactive protein (CRP). In men, an elevated CRP level was a significant determinant of CAD even after adjustment for CAD risk factors (OR, 3.1; 95% CI, 1.21 to 7.97). CMV seropositivity was associated with elevated CRP levels on multivariate analysis (P=0.006). In contrast, in women, CMV seropositivity was independently predictive of CAD (OR, 41.8; 95% CI, 4.12 to 423.74). CRP level in women with CAD was >25% higher than those without CAD, but the difference did not reach statistical significance. Importantly, compared with CMV Ab-/Tc- women, CAD prevalence was higher in Ab+/Tc- and Ab+/Tc+ (13% versus 68% and 64%, both P<0.005) but not in Ab-/Tc+ women (25%). There were no differences in age, smoking, diabetes, hypertension, and hypercholesterolemia among women with different types of immune responses to CMV infection. Conclusions-The mechanisms by which CMV predisposes to CAD in men and women may be different. In men, CMV appears to contribute to CAD risk, insofar as it predisposes to inflammation. In women, other mechanisms, possibly related to the type of immune response generated by the host, appear to be responsible for the proatherogenic effects of CMV. C1 Washington Hosp Ctr, Inst Cardiovasc Res, Washington, DC 20010 USA. Washington Hosp Ctr, MedStar Res Inst, Washington, DC 20010 USA. NCI, NIH, Bethesda, MD 20892 USA. NIH, Ctr Clin, Bethesda, MD 20892 USA. NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Div Epidemiol & Clin Applicat, NIH, Bethesda, MD 20892 USA. RP Epstein, SE (reprint author), Washington Hosp Ctr, Inst Cardiovasc Res, 110 Irving St NW,Suite 4B-1, Washington, DC 20010 USA. NR 27 TC 42 Z9 43 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 NOV 14 PY 2000 VL 102 IS 20 BP 2491 EP 2496 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 373NT UT WOS:000165296100017 PM 11076822 ER PT J AU Keller, K McCune, H Williams, C Muenke, M AF Keller, K McCune, H Williams, C Muenke, M TI Lobar holoprosencephaly in an infant born to a mother with classic phenylketonuria SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Letter ID MATERNAL PHENYLKETONURIA C1 Univ Florida, RC Philips Unit, Div Genet, Gainesville, FL 32610 USA. Natl Inst Hlth, Res Inst, Med Genet Branch Natl Human Genome, Bethesda, MD USA. RP Williams, C (reprint author), Univ Florida, RC Philips Unit, Div Genet, Box 100296, Gainesville, FL 32610 USA. NR 8 TC 8 Z9 8 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD NOV 13 PY 2000 VL 95 IS 2 BP 187 EP 188 DI 10.1002/1096-8628(20001113)95:2<187::AID-AJMG20>3.0.CO;2-G PG 2 WC Genetics & Heredity SC Genetics & Heredity GA 366VX UT WOS:000090027900020 PM 11078575 ER PT J AU Paasinen-Sohns, A Kielosto, M Kaariainen, E Eloranta, T Laine, A Janne, OA Birrer, MJ Holtta, E AF Paasinen-Sohns, A Kielosto, M Kaariainen, E Eloranta, T Laine, A Janne, OA Birrer, MJ Holtta, E TI c-Jun activation-dependent tumorigenic transformation induced paradoxically by overexpression or block of S-adenosylmethionine decarboxylase SO JOURNAL OF CELL BIOLOGY LA English DT Article DE cell transformation; c-Jun; JNK; S-adenosylmethionine decarboxylase; polyamines ID TERMINAL KINASE JNK; PROTEIN-KINASE; ORNITHINE DECARBOXYLASE; HA-RAS; ONCOGENIC TRANSFORMATION; SIGNALING PATHWAY; TUMOR PROMOTION; NIH3T3 CELLS; MAP KINASES; NUDE-MICE AB All mammalian cells absolutely require polyamines (putrescine, spermidine, and spermine) for growth. Here we show that the overexpression of cDNA for S-adenosylmethionine decarboxylase (AdoMetDC), the main regulatory enzyme in the biosynthesis of higher polyamines, induces transformation of rodent fibroblasts when expressed in the sense or the antisense orientation. Both transformants were able to induce invasive tumors in nude mice. Neither transformation was associated with activation of the mitogen-activated protein kinases Erk1 and Erk2. Instead, the AdoMet DC sense, but not antisense, transformants displayed constitutive activation of the c-Jun NH2-terminal, kinase (JNK) pathway. However, both transformations converged on persistent phosphorylation of endogenous c-Jun at Ser73. The phenotype of the AdoMetDC sense transformants was reversed by expression of dominant-negative mutants of SEK1 (MRK4), JNK1, and c-Jun (TAM-67), which were also found to impair cytokinesis. Similarly, TAM-67 reverted the morphology of the AdoMetDC-antisense expressors. This report is the first demonstration of a protein whose overexpression or block of synthesis can induce cell transformation. In addition, we show that the polyamine biosynthetic enzymes require c-Jun activation for eliciting their biological effects. C1 Univ Helsinki, Dept Pathol, Haartman Inst, FIN-00014 Helsinki, Finland. Univ Kuopio, Dept Biochem & Biotechnol, FIN-70211 Kuopio, Finland. Orion Corp Farmos, Expt Canc Res, FIN-20101 Turku, Finland. Univ Helsinki, Dept Physiol, Inst Biomed, FIN-00014 Helsinki, Finland. Univ Helsinki, Dept Clin Chem, FIN-00014 Helsinki, Finland. NCI, Div Canc Prevent & Control, Biomarkers & Prevent Res Branch, Rockville, MD 20850 USA. RP Holtta, E (reprint author), Univ Helsinki, Dept Pathol, Haartman Inst, POB 21,Haartmaninkatu 3, FIN-00014 Helsinki, Finland. NR 56 TC 19 Z9 22 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD NOV 13 PY 2000 VL 151 IS 4 BP 801 EP 809 DI 10.1083/jcb.151.4.801 PG 9 WC Cell Biology SC Cell Biology GA 375QF UT WOS:000165412200008 PM 11076965 ER PT J AU Alves, SE McEwen, BS Hayashi, S Korach, KS Pfaff, DW Ogawa, S AF Alves, SE McEwen, BS Hayashi, S Korach, KS Pfaff, DW Ogawa, S TI Estrogen-regulated progestin receptors are found in the midbrain raphe but not hippocampus of estrogen receptor alpha (ER alpha) gene-disrupted mice SO JOURNAL OF COMPARATIVE NEUROLOGY LA English DT Article DE serotonin; tryptophan hydroxylase; estrogen receptor beta; species difference ID MESSENGER-RNA EXPRESSION; OVARIAN-STEROID REGULATION; FEMALE RAT-BRAIN; CENTRAL-NERVOUS-SYSTEM; RHESUS MACAQUES; IMMUNOCYTOCHEMICAL LOCALIZATION; INSITU HYBRIDIZATION; DORSAL RAPHE; BETA; NEURONS AB Estrogen and progesterone may modulate serotonergic function through intracellular receptors, alpha (ER alpha) and/or beta (ER beta), and the progestin receptor (PR). Studies in macaque and rat suggest species differences in steroid action. Presently, we examined the mouse. To identify whether ER alpha is involved in estrogen induction of PR in midbrain raphe, we studied the ER alpha gene-disrupted (alpha ERKO) mouse. The hippocampus was examined as another estrogen/progestin-sensitive brain area reported to express ER alpha, ER beta, and PR. Female and male homozygous alpha ERKO and wildtype mice were gonadectomized and given estradiol benzoate or vehicle. Dual-label immunocytochemistry was performed for PR or ER alpha and the serotonin-synthesizing enzyme, tryptophan hydroxylase (TPH). Cells exhibiting PR immunoreactivity (PR-ir) or ER alpha -ir were observed in dorsal and median raphe and hippocampus in both sexes. No ER alpha -ir cells were observed in alpha ERKO brains. In raphe, PR-ir or ER alpha -ir often colocalized with TPH-ir. Thus, estrogen and progesterone may directly modulate gene expression in select serotonergic neurons via ER alpha and PR in female and male mice. Estrogen significantly increased the number of PR-ir cells, and the percentage of PR-ir cells colocalizing TPH-ir in both raphe nuclei, regardless of sex and genotype. Although less among alpha ERKO mice, the significant estrogen induction of PRs implicates the involvement of another ER, perhaps ERP. In hippocampus, distinct estrogen-induced PR-ir cells were observed only in wildtype animals, demonstrating an ER alpha -mediated event in this forebrain region. Collectively, these findings suggest that estrogen can regulate the expression of one gene (the PR) via multiple mechanisms, based upon brain region. (C) 2000 Wiley-Liss, Inc. C1 Rockefeller Univ, Neuroendocrinol Lab, New York, NY 10021 USA. Rockefeller Univ, Lab Behav & Mol Neurobiol, New York, NY 10021 USA. Tokyo Metropolitan Inst Neurosci, Dept Anat & Embryol, Tokyo 1838526, Japan. NIEHS, Receptor Biol Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Alves, SE (reprint author), Rockefeller Univ, Neuroendocrinol Lab, 1230 York Ave, New York, NY 10021 USA. OI Korach, Kenneth/0000-0002-7765-418X FU NIMH NIH HHS [MH59251, MH62147]; NINDS NIH HHS [NS07080] NR 50 TC 64 Z9 65 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0021-9967 J9 J COMP NEUROL JI J. Comp. Neurol. PD NOV 13 PY 2000 VL 427 IS 2 BP 185 EP 195 DI 10.1002/1096-9861(20001113)427:2<185::AID-CNE2>3.0.CO;2-G PG 11 WC Neurosciences; Zoology SC Neurosciences & Neurology; Zoology GA 368HR UT WOS:000090101300002 PM 11054687 ER PT J AU Dey, BR Furlanetto, RW Nissley, P AF Dey, BR Furlanetto, RW Nissley, P TI Suppressor of cytokine signaling (SOCS)-3 protein interacts with the insulin-like growth factor-I receptor SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE yeast two-hybrid; phospho-SOCS-3; insulin receptor ID MESSENGER-RIBONUCLEIC-ACID; SOCS-3; EXPRESSION; CLONING; FAMILY; DIFFERENTIATION; ACTIVATION; INHIBITOR; BINDING; CIS3 AB SOCS proteins are a class of proteins that are negative regulators of cytokine receptor signaling via the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. In a yeast two-hybrid screen of a human fetal brain library, we have previously identified SOCS-2 as a binding partner of the activated IGF-I receptor (IGFIR). To test whether or not SOCS-3 also binds to the IGFIR, we cloned human SOCS-3 by reverse transcription-polymerase chain reaction from human skeletal muscle mRNA. SOCS-3 mRNA was expressed in many human fetal and adult tissues and in some human cancer cell lines (Hela, A549 pulmonary adenocarcinoma and G361 human melanoma). We found that human SOCS-3 protein interacts directly with the cytoplasmic domains of the activated IGFIR and the insulin receptor (IR) in the yeast two-hybrid assay. In GST-SOCS-3 pull-down experiments using IGFIR from mammalian cells and in immunoprecipitation experiments in which IGFIR and FLAG-SOCS-3 were transiently expressed in human embryonic kidney 293 cells, we found that SOCS-3 interacts constitutively with IGFIR in vitro and in intact cells. Unlike SOCS-2, hSOCS-3 was phosphorylated on tyrosines in response to IGF-I addition to 293 cells. We conclude that SOCS-3 binds to the IGFIR and may be a direct substrate for the receptor tyrosine kinase. (C) 2000 Academic Press. C1 NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. Univ Rochester, Sch Med & Dent, Dept Pediat, Strong Childrens Res Ctr, Rochester, NY 14642 USA. RP Nissley, P (reprint author), NCI, Metab Branch, NIH, Bldg 10,Rm 4N115, Bethesda, MD 20892 USA. NR 23 TC 66 Z9 72 U1 0 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD NOV 11 PY 2000 VL 278 IS 1 BP 38 EP 43 DI 10.1006/bbrc.2000.3762 PG 6 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 375QP UT WOS:000165413000007 PM 11071852 ER PT J AU Mori-Aoki, A Pietrarelli, M Nakazato, M Caturegli, P Kohn, LD Suzuki, K AF Mori-Aoki, A Pietrarelli, M Nakazato, M Caturegli, P Kohn, LD Suzuki, K TI Class II transactivator suppresses transcription of thyroid-specific genes SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE MHC; thyroid; autoimmunity; transcription factors; antigen presentation; and gene regulation ID COMPLEX CLASS-II; THYROTROPIN RECEPTOR GENE; Y-BOX PROTEIN; GRAVES-DISEASE; STIMULATING HORMONE; SYMPORTER GENE; FRTL-5 CELLS; IN-VIVO; EXPRESSION; CIITA AB Class II transactivator (CIITA) is the master regulator of MHC class II genes, and mediates their induction by interferon gamma (IFN gamma). To study the role of CIITA in modulating the expression of thyroid-specific genes, we cloned the full-length rat CIITA and use it to transfect a rat thyroid cell line. We found that only one type of CIITA, type TV, is induced in thyroid cells upon IFN gamma stimulation, and that CIITA is capable not only of inducing the expression of MHC genes in the thyroid, but also of differentially suppressing the expression of thyroid-specific genes. These findings suggest new avenues for the development of thyroid autoimmune diseases. (C) 2000 Academic Press. C1 NIDDKD, Metab Dis Branch, Cell Regulat Sect, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Bethesda, MD 20892 USA. RP Suzuki, K (reprint author), Washington Hosp Ctr, MedStar Res Inst, Mol Endocrinol Lab, 108 Irving St,Hyman Bldg,Room 221 NW, Washington, DC 20010 USA. NR 32 TC 13 Z9 13 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD NOV 11 PY 2000 VL 278 IS 1 BP 58 EP 62 DI 10.1006/bbrc.2000.3769 PG 5 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 375QP UT WOS:000165413000010 PM 11071855 ER PT J AU Watanabe, S Take, H Takeda, K Yu, ZX Iwata, N Kajigaya, S AF Watanabe, S Take, H Takeda, K Yu, ZX Iwata, N Kajigaya, S TI Characterization of the CIN85 adaptor protein and identification of components involved in CIN85 complexes SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE BLNK; c-Cbl; CMS; CD2AP; coiled-coils ID RECEPTOR SIGNAL-TRANSDUCTION; PROTOONCOGENE C-CBL; B-CELL DEVELOPMENT; NEGATIVE REGULATOR; ANTIGEN RECEPTOR; TYROSINE KINASE; LINKER PROTEIN; COILED COILS; BLNK; ACTIVATION AB CIN85 is an 85-kDa adaptor protein whose functions in signaling pathways are presently unknown. Using the yeast two-hybrid screen, the B cell linker protein (BLNK) was identified as a binding partner of CIN85. Coimmunoprecipitation experiments using mammalian cells revealed that CIN85 directly bound to BLNK through its SH3 domains. Immunostaining analysis showed that CIN85 and BLNK were colocalized in the cytoplasm. These results indicate a potential role of CIN85 in the B cell receptor-mediated signaling pathway. It was also found that Crk-I, Crk-II, p130(Cas), p85-PI3K, Grb2, and Sos1 were components of CIN85 complexes. CIN85 interacted with itself through its coiled-coil region, resulting in formation of a tetramer. Both the coiled-coil region and SH3 domains of CIN85 were responsible for its subcellular localization. Our data suggest that CIN85 may serve for regulation of various signaling events through formation of its diverse complexes. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Kajigaya, S (reprint author), NHLBI, Hematol Branch, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 26 TC 62 Z9 67 U1 1 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD NOV 11 PY 2000 VL 278 IS 1 BP 167 EP 174 DI 10.1006/bbrc.2000.3760 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 375QP UT WOS:000165413000026 PM 11071869 ER PT J AU Morens, DM AF Morens, DM TI Snow and the Broad Street pump: a rediscovery SO LANCET LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Morens, DM (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 5 TC 2 Z9 2 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 NOV 11 PY 2000 VL 356 IS 9242 BP 1688 EP 1689 DI 10.1016/S0140-6736(05)70399-8 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 372TB UT WOS:000165249200056 PM 11089854 ER PT J AU Patterson, LJ Peng, B Abimiku, AG Aldrich, K Murty, L Markham, PD Kalyanaraman, VS Alvord, WG Tartaglia, J Franchini, G Robert-Guroff, M AF Patterson, LJ Peng, B Abimiku, AG Aldrich, K Murty, L Markham, PD Kalyanaraman, VS Alvord, WG Tartaglia, J Franchini, G Robert-Guroff, M TI Cross-protection in NYVAC-HIV-1-immunized/HIV-2-challenged but not in NYVAC-HIV-2-immunized/SHIV-challenged rhesus macaques SO AIDS LA English DT Article DE cross-protection; HIV-1; HIV-2; SHIV; rhesus macaques; poxvirus; AIDS vaccine ID IMMUNODEFICIENCY-VIRUS TYPE-1; RECOMBINANT VACCINIA VIRUSES; CYTOTOXIC T-LYMPHOCYTES; ENVELOPE GLYCOPROTEIN; HIV-1 INFECTION; EXPRESSION VECTOR; MONKEYS; CELLS; NEUTRALIZATION; IMMUNIZATION AB Objectives: Immunization with attenuated poxvirus-HIV-1 recombinants followed by protein boosting had protected four of eight rhesus macaques from HIV-2(SBL6669) challenge. The present study was designed to confirm this result and to conduct the reciprocal cross-protection experiment. Methods: Twenty-four macaques were primed with NYVAC (a genetically attenuated Copenhagen vaccinia strain) recombinants with HIV-1 and HIV-2 env and gag-pol or NYVAC vector alone and boosted with homologous, oligomeric gp160 proteins or adjuvant only. Binding and neutralizing antibodies, cytotoxic T-lymphocytes (CTL) and CD8 T cell antiviral activity (CD8AA) were evaluated. One half of each immunization and control group were intravenously challenged with SHIVHXB2 the other half was challenged with HIV-2(SBL6669), Protective outcome was assessed by monitoring virus isolation, proviral DNA and plasma viral RNA. Results: Both immunization groups developed homologous binding antibodies; however, homologous neutralizing antibodies were only observed in NYVAC-HIV-2-immunized macaques. While no cross-reactive neutralizing antibodies were detected, both immunization groups displayed cross-reactive CTL. Significant CD8AA was observed for only one NYVAC-HIV-2-immunized macaque. Virological assessments verified that both NYVAC-HIV-1 and NYVAC-HIV-2 immunization significantly reduced viral burdens and partially protected against HIV-2 challenge, although cross-protection was not at the level that had been previously reported. Humoral antibody and/or CTL and CD8AA were associated with protection against homologous HIV-2 challenge, while cellular immune responses seemed more important for cross-protection. No significant protection was observed in the SHIV-challenged macaques, although NYVAC-HIV-1 immunization resulted in significantly lower viral burdens compared with controls. Conclusions: Further delineation of cross-reactive mechanisms may aid in the development of a broadly protective vaccine. (C) 2000 Lippincott Williams & Wilkins. C1 NCI, Basic Res Lab, Bethesda, MD 20892 USA. Univ Maryland, Inst Human Virol, Baltimore, MD 21201 USA. Adv Biosci Labs Inc, Kensington, NSW, Australia. NCI, Frederick Canc Res & Dev Ctr, Data Management Serv, Frederick, MD USA. Virogenet Corp, Troy, NY 12180 USA. RP Robert-Guroff, M (reprint author), NCI, Basic Res Lab, 41 Lib Dr MSC 5055,Bldg 41,Room D804, Bethesda, MD 20892 USA. NR 42 TC 7 Z9 8 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD NOV 10 PY 2000 VL 14 IS 16 BP 2445 EP 2455 DI 10.1097/00002030-200011100-00005 PG 11 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 373ZW UT WOS:000165320800005 PM 11101054 ER PT J AU Ruxrungtham, K Suwanagool, S Tavel, JA Cheunyam, M Kroon, E Ubolyam, S Buranapraditkun, S Techasathit, W Li, YM Emery, S Davey, RT Fosdick, L Kunanusont, C Lane, HC Phanuphak, P AF Ruxrungtham, K Suwanagool, S Tavel, JA Cheunyam, M Kroon, E Ubolyam, S Buranapraditkun, S Techasathit, W Li, YM Emery, S Davey, RT Fosdick, L Kunanusont, C Lane, HC Phanuphak, P CA Vanguard Study Grp TI A randomized, controlled 24-week study of intermittent subcutaneous interleukin-2 in HIV-1 infected patients in Thailand SO AIDS LA English DT Article DE Asia; CD4; immune-based therapy; Thailand ID HUMAN-IMMUNODEFICIENCY-VIRUS; INTRAVENOUS INTERLEUKIN-2; CONTROLLED TRIAL; LYMPHOCYTES; IMPACT AB Objectives: To assess the immunological and virological effects, safety profile and feasibility of subcutaneous interleukin-2 (sCIL-2) therapy in an HIV-infected Thai population. Design: Seventy-two patients with baseline CD4 cell count of greater than or equal to 350 x 10(6)/L and no history of opportunistic infection were randomized to receive antiretroviral therapy plus scIL-2 (scIL-2 group) or antiretroviral therapy alone (control group). scIL-2 was administered at one of three doses for at least 24 weeks. The main measure of treatment efficacy was change in CD4 cell count. Results: The time-weighted mean change in CD4 cell count from baseline to week 24 was + 252 x 10(6)/l for the scIL-2 group compared with + 42 x 10(6)/l for the control group (P < 0.0001). Changes in plasma HIV RNA were not significantly different between the groups over the same time period: there was a 0.83 log(10) copies/ml decrease for the scIL-2 group and a 0.70 log copies/ml decrease for the control group (P = 0.362). Conclusions: This study provides the most extensive experience of scIL-2 therapy in HIV-1 infected women and Asians, and demonstrates the immunological efficacy, tolerability and feasability of scIL-2 therapy in this population. Data from this study were instrumental in guiding the selection of the scIL-2 dosing regimen for ongoing phase III trials. (C) 2000 Lippincott Williams & Wilkins. C1 Thai Red Cross, AIDS Res Ctr, HIV Netherlands Australia Thailand Res Collaborat, Bangkok 10330, Thailand. Chulalongkorn Univ, Fac Med, Dept Med, Bangkok 10330, Thailand. Mahidol Univ, Siriraj Hosp, Bangkok 10700, Thailand. NIAID, NIH, Bethesda, MD 20892 USA. Univ Amsterdam, Acad Med Ctr, Dept Internal Med, Natl AIDS Therapy Evaluat Ctr, NL-1105 AZ Amsterdam, Netherlands. Chulalongkorn Univ, Fac Med, Dept Microbiol, Bangkok 10330, Thailand. Natl Ctr HIV Epidemiol & Clin Res, Sydney, NSW, Australia. Univ Minnesota, Coordinating Ctr Biometr Res, Minneapolis, MN 55455 USA. Minist Publ Hlth, Div Communicable Dis, Div AIDS, Bangkok, Thailand. NCHECR, Sydney, NSW, Australia. NATEC, Amsterdam, Netherlands. HIV NAT, Bangkok, Thailand. MOPH, Bangkok, Thailand. RP Ruxrungtham, K (reprint author), Thai Red Cross, AIDS Res Ctr, HIV Netherlands Australia Thailand Res Collaborat, 104 Rajadamri Rd,Lumpini, Bangkok 10330, Thailand. NR 12 TC 32 Z9 32 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD NOV 10 PY 2000 VL 14 IS 16 BP 2509 EP 2513 DI 10.1097/00002030-200011100-00013 PG 5 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 373ZW UT WOS:000165320800013 PM 11101062 ER PT J AU Grene, E Pinto, LA Kwak-Kim, JYH Giorgi, JV Landay, AL Kessler, HA Beer, AE Shearer, GM AF Grene, E Pinto, LA Kwak-Kim, JYH Giorgi, JV Landay, AL Kessler, HA Beer, AE Shearer, GM TI Increased levels of anti-CCR5 antibodies in sera from individuals immunized with allogeneic lymphocytes SO AIDS LA English DT Letter ID CHEMOKINE RECEPTOR; INFECTION; HIV-1; CCR5; INDUCTION C1 NCI, Expt Immunol Branch, Bethesda, MD 20892 USA. Finch Univ Hlth Sci Chicago Med Sch, Div Reprod Med, Chicago, IL USA. Univ Calif Los Angeles, Sch Med, Los Angeles, CA 90024 USA. Rush Presbyterian St Lukes Med Ctr, Dept Microbiol Immunol, Chicago, IL 60612 USA. Rush Presbyterian St Lukes Med Ctr, Dept Med, Chicago, IL 60612 USA. RP Grene, E (reprint author), NCI, Expt Immunol Branch, Bethesda, MD 20892 USA. NR 10 TC 2 Z9 2 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 NOV 10 PY 2000 VL 14 IS 16 BP 2627 EP 2628 DI 10.1097/00002030-200011100-00035 PG 2 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 373ZW UT WOS:000165320800035 PM 11101084 ER PT J AU Suzuki, M Youle, RJ Tjandra, N AF Suzuki, M Youle, RJ Tjandra, N TI Structure of Bax: Coregulation of dimer formation and intracellular localization SO CELL LA English DT Article ID PROGRAMMED CELL-DEATH; CYTOCHROME-C RELEASE; ORIENTED MACROMOLECULES; BACKBONE DYNAMICS; DIPOLAR COUPLINGS; CHEMICAL-SHIFT; BCL-2 FAMILY; BH3 DOMAIN; APOPTOSIS; MITOCHONDRIA AB Apoptosis is stimulated by the insertion of Bax from the cytosol into mitochondrial membranes. The solution structure of Bax, including the putative transmembrane domain at the C terminus, was determined in order to understand the regulation of its subcellular location. Bax consists of 9 alpha helices where the assembly of helices alpha1 through alpha8 resembles that of the apoptosis inhibitor, Bcl-x(L). The C-terminal alpha9 helix occupies the hydrophobic pocket proposed previously to mediate heterodimer formation and bioactivity of opposing members of the Bcl-2 family. The Bax structure shows that the orientation of helix alpha9 provides simultaneous control over its mitochondrial targeting and dimer formation. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. NINCDS, Biochem Sect, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Tjandra, N (reprint author), NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. NR 41 TC 683 Z9 708 U1 3 U2 21 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD NOV 10 PY 2000 VL 103 IS 4 BP 645 EP 654 DI 10.1016/S0092-8674(00)00167-7 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 374MB UT WOS:000165348000009 PM 11106734 ER PT J AU Peirce, M Metzger, H AF Peirce, M Metzger, H TI Detergent-resistant microdomains offer no refuge for proteins phosphorylated by the IgE receptor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FC-EPSILON-RI; BASOPHILIC LEUKEMIA-CELLS; HIGH-AFFINITY RECEPTOR; IMMUNOGLOBULIN-E; TYROSINE PHOSPHORYLATION; SIGNAL-TRANSDUCTION; MEMBRANE DOMAINS; ACTIVATION; PHOSPHATASES; MECHANISM AB When the high affinity receptor for IgE and related receptors become aggregated, they emigrate to specialized microdomains of the plasma membrane that are enriched in certain lipids and lipid-anchored proteins. Among the latter are the kinases that initiate signaling cascade(s) by phosphorylating the receptors. In studying the IgE receptor, we explored whether, in addition to their potential role in enhancing the initiation of signaling by the kinase(s), the microdomains might augment the stimulation by excluding phosphatases. In vitro assessment of phosphatase activity, using either a relevant or irrelevant substrate, suggested that the microdomains were deficient in phosphatase activity, but, in vivo, proteins confined to the microdomains were found to be no less vulnerable to dephosphorylation than those outside such domains. In the course of our experiments, we observed that the procedures routinely used to isolate the detergent-resistant domains dissociated the receptor for IgE, thereby artificially accentuating the observed preferential distribution of phosphorylated subunits in the microdomains. C1 NIAMS, Arthritis & Rheumatism Branch, Sect Chem Immunol, NIH, Bethesda, MD 20892 USA. RP Metzger, H (reprint author), NIAMS, Arthritis & Rheumatism Branch, Sect Chem Immunol, NIH, Bldg 10,Rm 9N-228,10 Ctr Dr,MSC-1820, Bethesda, MD 20892 USA. NR 42 TC 24 Z9 24 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 34976 EP 34982 DI 10.1074/jbc.M005819200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800022 PM 10956655 ER PT J AU Drugan, JK Rogers-Graham, K Gilmer, T Campbell, S Clark, GJ AF Drugan, JK Rogers-Graham, K Gilmer, T Campbell, S Clark, GJ TI The Ras/p120 GTPase-activating protein (GAP) interaction is regulated by the p120 GAP pleckstrin homology domain SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BETA-GAMMA-SUBUNITS; BRUTON TYROSINE KINASE; HIGH-AFFINITY BINDING; RAS-GTPASE; PHOSPHOINOSITIDE BINDING; PH DOMAIN; TRANSFORMATION; IDENTIFICATION; SPECIFICITY; GENE AB Pleckstrin homology domains are structurally conserved functional domains that can undergo both protein/protein and protein/lipid interactions. Pleckstrin homology domains can mediate inter- and intra-molecular binding events to regulate enzyme activity. They occur in numerous proteins including many that interact with Pas superfamily members, such as p120 GAP, The pleckstrin homology domain of p120 GAP is located in the NH2-terminal, noncatalytic region of p120 GAP. Overexpression of the noncatalytic domains of p120 GAP may modulate Pas signal transduction pathways. Here, we demonstrate that expression of the isolated pleckstrin homology domain of p120 GAP specifically inhibits Pas-mediated signaling and transformation but not normal cellular growth. Furthermore, we show that the pleckstrin homology domain binds the catalytic domain of p120 GAP and interferes with the Ras/GAP interaction. Thus, we suggest that the pleckstrin homology domain of p120 GAP may specifically regulate the interaction of Pas with p120 GAP via competitive intramolecular binding. C1 NCI, Dept Cell & Canc Biol, NIH, Rockville, MD 20850 USA. Univ N Carolina, Dept Pharmacol, Chapel Hill, NC 27599 USA. Glaxo Wellcome Inc, Dept Cell Biol, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Biochem, Chapel Hill, NC 27599 USA. RP Clark, GJ (reprint author), NCI, Dept Cell & Canc Biol, NIH, 9610 Med Ctr Dr, Rockville, MD 20850 USA. FU NCI NIH HHS [CA70308, CA64569, CA72644-10] NR 42 TC 26 Z9 26 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35021 EP 35027 DI 10.1074/jbc.M004386200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800028 PM 10954709 ER PT J AU Yan, ZJ Jetten, AM AF Yan, ZJ Jetten, AM TI Characterization of the repressor function of the nuclear orphan receptor retinoid receptor-related testis-associated receptor/germ cell nuclear factor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID STEROID-HORMONE RECEPTORS; DNA-BINDING; ESTROGEN-RECEPTOR; RESPONSE ELEMENT; TRANSCRIPTIONAL ACTIVATION; CHROMOSOMAL LOCALIZATION; DIFFERENTIAL EXPRESSION; SILENCING MEDIATOR; CO-REPRESSOR; N-COR AB Retinoid receptor-related testis-associated receptor (RTR)/germ cell nuclear factor is a nuclear orphan receptor that plays an important role in the control of gene expression during early embryonic development and gametogenesis. It has been shown to repress transcriptional activation, In this study, we further characterize this repressor function. We demonstrate that RTR can suppress the transcriptional activation induced by the estrogen receptor related-receptor alpha1 through its response element. The latter is at least in part due to competition for binding to the same response element. In addition, RTR inhibits basal transcriptional activation, indicating that it functions as an active repressor. Mammalian two-hybrid analyses showed that RTR interacts with the co-repressor nuclear co-repressor (N-CoR) but is unable to interact with the co-repressor SMRT or RIP140. Pull-down analyses with glutathione S-transferase-RTR fusion protein demonstrated that RTR physically interacts with N-CoR in vitro, suggesting a potential role for N-CoR in the transcriptional repression by RTR. To identify the regions in RTR essential for the binding of RTR to N-CoR, the effect of various deletion and point mutations on this interaction was examined. This analysis revealed that this interaction requires the hinge domain, helix 3 as well as the helix 12 region of RTR. The residues Ser(246)-Tyr(247) in the hinge domain, Lys(318) in helix 3, and Lys(489)-Thr(490) in helix 12 are identified as being critical in this interaction. Our results demonstrate that RTR can function as an active transcriptional repressor and that this repression can be mediated through interactions with the corepressor N-CoR. We show that this interaction exhibits several characteristics unique to RTR. Through its repressor function, RTR can suppress the induction of transcriptional activation by other nuclear receptors. These repressor activities may provide important mechanisms by which RTR regulates gene expression during development and gametogenesis. C1 NIEHS, Pulm Pathobiol Lab, Cell Biol Sect, NIH, Res Triangle Pk, NC 27709 USA. RP Jetten, AM (reprint author), NIEHS, Pulm Pathobiol Lab, Cell Biol Sect, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. OI Jetten, Anton/0000-0003-0954-4445 NR 56 TC 35 Z9 35 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35077 EP 35085 DI 10.1074/jbc.M005566200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800035 PM 10940306 ER PT J AU Kim, HY Akbar, M Lau, A Edsall, L AF Kim, HY Akbar, M Lau, A Edsall, L TI Inhibition of neuronal apoptosis by docosahexaenoic acid (22 : 6n-3) - Role of phosphatidylserine in antiapoptotic effect SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID POLYUNSATURATED FATTY-ACIDS; C6 GLIOMA-CELLS; ARACHIDONIC-ACID; RETINAL PHOTORECEPTORS; SIGNAL-TRANSDUCTION; PLASMA-MEMBRANE; RAF-1 ACTIVITY; IN-VITRO; SURVIVAL; DEATH AB Enrichment of Neuro 2A cells with docosahexaenoic acid (22:6n-3) decreased apoptotic cell death induced by serum starvation as evidenced by the reduced DNA fragmentation and caspase-3 activity. The protective effect of 22:6n-3 became evident only after at least 24 h of enrichment before serum starvation and was potentiated as a function of the enrichment period, During enrichment 22:6n-3 incorporated into phosphatidylserine (PS) steadily, resulting in a significant increase in the total PS content. Similar treatment with oleic acid (18:1n-9) neither altered PS content nor resulted in protective effect. Hindering PS accumulation by enriching cells in a serine-free medium diminished the protective effect of 22:6n-3. Membrane translocation of Raf-l was significantly enhanced by 22:6n-3 enrichment in Neuro 2A cells. Consistently, in vitro biomolecular interaction between PS/phosphatidylethanolamine /phosphatidylcholine liposomes, and Raf-l increased in a PS concentration-dependent manner. Collectively, enrichment of neuronal cells with 22:6n-3 increases the PS content and Raf-l translocation, down-regulates caspase-3 activity, and prevents apoptotic cell death. Both the antiapoptotic effect of 22:6n-3 and Raf-l translocation are sensitive to 22:6n-3 enrichment-induced PS accumulation, strongly suggesting that the protective effect of 22:6n-3 may be mediated at least in part through the promoted accumulation of PS in neuronal membranes. C1 NIAAA, LMBB, Sect Mass Spectrometry, NIH, Rockville, MD 20852 USA. RP Kim, HY (reprint author), NIAAA, LMBB, Sect Mass Spectrometry, NIH, 12420 Parklawn Dr, Rockville, MD 20852 USA. NR 57 TC 185 Z9 200 U1 1 U2 9 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35215 EP 35223 DI 10.1074/jbc.M004446200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800053 PM 10903316 ER PT J AU Guschin, D Geiman, TM Kikyo, N Tremethick, DJ Wolffe, AP Wade, PA AF Guschin, D Geiman, TM Kikyo, N Tremethick, DJ Wolffe, AP Wade, PA TI Multiple ISWI ATPase complexes from Xenopus laevis - Functional conservation of an ACF/CHRAC homolog SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID MAJOR DEVELOPMENTAL TRANSITION; CHROMATIN-REMODELING FACTORS; CELL-FREE SYSTEM; HISTONE OCTAMER; GENE-EXPRESSION; DNA-REPAIR; SWI-SNF; NUCLEOSOME; OOCYTES; TRANSCRIPTION AB The nucleosomal ATPase ISWI is the catalytic subunit of several protein complexes that either organize or perturb chromatin structure in vitro. This work reports the cloning and biochemical characterization of a Xenopus ISWI homolog. Surprisingly, whereas we find four complex forms of ISWI in egg extracts, we find no functional homolog of NURF. One of these complexes, xACF, consists of ISWI, Acf1, and a previously uncharacterized protein of 175 kDa. Like both ACF and CHRAC, this complex organizes randomly deposited histones into a regularly spaced array. The remaining three forms include two novel ISWI complexes distinct from known ISWI complexes plus a histone-dependent ATPase complex. This comprehensive biochemical characterization of ISWI underscores the evolutionary conservation of the ACF/CHRAC family. C1 NICHD, Mol Embryol Lab, NIH, Bethesda, MD 20892 USA. Australian Natl Univ, John Curtin Sch Med Res, Canberra, ACT 2601, Australia. RP Wade, PA (reprint author), Emory Univ, Dept Pathol, Woodruff Mem Res Bldg,Rm 7105-B,1639 Pierce Dr, Atlanta, GA 30322 USA. NR 61 TC 57 Z9 58 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35248 EP 35255 DI 10.1074/jbc.M006041200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800057 PM 10942776 ER PT J AU Hoskins, JR Kim, SY Wickner, S AF Hoskins, JR Kim, SY Wickner, S TI Substrate recognition by the ClpA chaperone component of ClpAP protease SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ATP-DEPENDENT PROTEASES; HEAT-SHOCK PROTEINS; N-END RULE; ESCHERICHIA-COLI; CRYSTAL-STRUCTURE; MU-TRANSPOSASE; REGULATORY SUBUNITS; MOLECULAR CHAPERONE; DNA-REPLICATION; QUALITY-CONTROL AB ClpA, a member of the Clp/Hsp100 ATPase family, is a molecular chaperone and regulatory component of Clp-AP protease, We explored the mechanism of protein recognition by ClpA using a high affinity substrate, RepA, which is activated for DNA binding by ClpA and degraded by ClpAP, By characterizing RepA derivatives with N- or C-terminal deletions, we found that the N-terminal portion of RepA is required for recognition. More precisely, RepA derivatives lacking the N-terminal 5 or 10 amino acids are degraded by ClpAP at a rate similar to full-length RepA, whereas RepA derivatives lacking 15 or 20 amino acids are degraded much more slowly. Thus, ClpA recognizes an N-terminal signal in RepA beginning in the vicinity of amino acids 10-15, Moreover, peptides corresponding to RepA amino acids 4-13 and 1-15 inhibit interactions between ClpA and RepA We constructed fusions of RepA and green fluorescent protein, a protein not recognized by ClpA, and found that the N-terminal 15 amino acids of RepA are sufficient to target the fusion protein for degradation by ClpAP. However, fusion proteins containing 46 or 70 N-terminal amino acids of RepA are degraded more efficiently in vitro and are noticeably stabilized in vivo in clpA Delta and clpP Delta strains compared with wild type. C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Wickner, S (reprint author), NCI, Mol Biol Lab, NIH, Bldg 37,Rm 2D19,37 Convent Dr,MSC37-4255, Bethesda, MD 20892 USA. NR 53 TC 53 Z9 54 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35361 EP 35367 DI 10.1074/jbc.M006288200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800071 PM 10952988 ER PT J AU Zhang, J Billingsley, ML Kincaid, RL Siraganian, RP AF Zhang, J Billingsley, ML Kincaid, RL Siraganian, RP TI Phosphorylation of Syk activation loop tyrosines is essential for Syk function - An in vivo study using a specific anti-Syk activation loop phosphotyrosine antibody SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FC-EPSILON-RI; AFFINITY IGE RECEPTOR; BASOPHILIC LEUKEMIA-CELLS; SIGNAL-TRANSDUCTION; KINASE P72(SYK); SH2 DOMAINS; MAST-CELLS; NEGATIVE REGULATION; IMMUNOGLOBULIN-E; FAMILY KINASES AB Syk is an important protein-tyrosine kinase in immunoreceptor signaling. Fc epsilon RI aggregation in mast cells induces tyrosine phosphorylation and increased enzymatic activity of Syk. The two adjacent tyrosines in the Syk activation loop are thought to be important for the propagation of Fc epsilon RI signaling. To evaluate the phosphorylation of these tyrosines in vivo and further understand the relationship of Syk tyrosine phosphorylation with its function, an antibody was developed specific for phosphorylated tyrosines in the activation loop of Syk. Fc epsilon RI aggregation on mast cells induced the phosphorylation of both tyrosine residues of the activation loop. The kinase activity of Syk played the major role in phosphorylating its activation loop tyrosines both in vivo and in vitro. In Fc epsilon RI-stimulated mast cells, the total Syk tyrosine phosphorylation paralleled the phosphorylation of its activation loop tyrosines and downstream propagation of signals for histamine release. In contrast,:the cell surface binding of anti-ganglioside monoclonal antibody AA4 induced only strong general tyrosine: phosphorylation of Syk and minimal histamine release and weak phosphorylation of activation loop tyrosines. These results demonstrate that phosphorylation:of the activation loop tyrosines is important for mediating receptor signaling and is a better marker of Syk function than is total Syk tyrosine phosphorylation. C1 NIDCR, RAST Sect, OIIB, NIH, Bethesda, MD 20892 USA. Penn State Univ, Coll Med, Dept Pharmacol, Hershey, PA 17033 USA. RP Zhang, J (reprint author), NIDCR, RAST Sect, OIIB, NIH, Bldg 10,Rm 1N106, Bethesda, MD 20892 USA. FU NIDCR NIH HHS [N01-DE-62614] NR 49 TC 102 Z9 103 U1 0 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35442 EP 35447 DI 10.1074/jbc.M004549200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800081 PM 10931839 ER PT J AU Kronfeld, I Kazimirsky, G Lorenzo, PS Garfield, SH Blumberg, PM Brodie, C AF Kronfeld, I Kazimirsky, G Lorenzo, PS Garfield, SH Blumberg, PM Brodie, C TI Phosphorylation of protein kinase C delta on distinct tyrosine residues regulates specific cellular functions SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HIGH-AFFINITY RECEPTOR; PHORBOL ESTER; PKC-DELTA; SIGNAL-TRANSDUCTION; GLUTAMINE-SYNTHETASE; GLIAL-CELLS; ACTIVATION; EXPRESSION; DOMAINS; SRC AB Protein kinase C delta (PKC delta) inhibits proliferation and decreases expression of the differentiation marker glutamine synthetase (GS) in C6 glioma cells. Here, we report that distinct, specific tyrosine residues on PKC delta are involved in these two responses. Transfection of cells with PKC delta mutated at tyrosine 155 to phenylalanine caused enhanced proliferation in response to 12-phorbol la-myristate 13-acetate, whereas GS expression resembled that for the PKC delta wild-type transfectant. Conversely, transfection with PKC delta mutated at tyrosine 187 to phenylalanine resulted in increased expression of GS, whereas the rate of proliferation resembled that of the PKC delta wild-type transfectant. The tyrosine phosphorylation of PKC delta and the decrease in GS expression induced by platelet-derived growth factor (PDGF) were abolished by the Src kinase inhibitors PP1 and PP2. In response to PDGF, Fyn associated with PKC delta via tyrosine 187. Finally, overexpression of dominant negative Fyn abrogated the decrease in GS expression and reduced the tyrosine phosphorylation of PKC delta induced by PDGF. We conclude that the tyrosine phosphorylation of PKC delta and its association with tyrosine kinases may be an important point of divergence in PKC signaling. C1 Bar Ilan Univ, Fac Life Sci, Gonda Goldschmied Med Diag Res Ctr, IL-52900 Ramat Gan, Israel. NCI, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. NCI, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Brodie, C (reprint author), Bar Ilan Univ, Fac Life Sci, Gonda Goldschmied Med Diag Res Ctr, IL-52900 Ramat Gan, Israel. NR 43 TC 87 Z9 88 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35491 EP 35498 DI 10.1074/jbc.M005991200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800087 PM 10945993 ER PT J AU Kumaraswamy, E Malykh, A Korotkov, KV Kozyavkin, S Hu, Y Kwon, SY Moustafa, ME Carlson, BA Berry, MJ Lee, BJ Hatfield, DL Diamond, AM Gladyshev, VN AF Kumaraswamy, E Malykh, A Korotkov, KV Kozyavkin, S Hu, Y Kwon, SY Moustafa, ME Carlson, BA Berry, MJ Lee, BJ Hatfield, DL Diamond, AM Gladyshev, VN TI Structure-expression relationships of the 15-kDa selenoprotein gene - Possible role of the protein in cancer etiology SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NICOTINIC-ACID HYDROXYLASE; SELENOCYSTEINE-CONTAINING PROTEINS; CLOSTRIDIUM-BARKERI; GLUTATHIONE-PEROXIDASE; MAMMALIAN-CELLS; MESSENGER-RNA; SELENIUM; SEQUENCE; MOUSE; DEIODINASE AB Selenium has been implicated in cancer prevention, but the mechanism and possible involvement of seleno-proteins in this process are not understood. To elucidate whether the 15-kDa selenoprotein may play a role in cancer etiology, the complete sequence of the human 15-kDa protein gene was determined, and various characteristics associated with expression of the protein were examined in normal and malignant cells and tissues. The 51-kilobase pair gene for the 15-kDa selenoprotein consisted of five exons and four introns and was localized on chromosome 1p31, a genetic locus commonly mutated or deleted in human cancers. Two stem-loop structures resembling selenocysteine insertion sequence elements were identified in the 3'-untranslated region of the gene, and only one of these was functional. Two alleles in the human 15-kDa protein gene were identified that differed by two single nucleotide polymorphic sites that occurred within the selenocysteine insertion sequence-like structures. These S'-untranslated region polymorphisms resulted in changes in selenocysteine incorporation into protein and responded differently to selenium supplementation. Human and mouse 15-kDa selenoprotein genes manifested the highest level of expression in prostate, liver, kidney, testis, and brain, and the level of the selenoprotein was reduced substantially in a malignant prostate cell line and in hepatocarcinoma. The expression pattern of the 15-kDa protein in normal and malignant tissues, the occurrence of polymorphisms associated with protein expression, the role of selenium in differential regulation of polymorphisms, and the chromosomal location of the gene may be relevant to a role of this protein in cancer. C1 Univ Nebraska, Dept Biochem, Beadle Ctr, Lincoln, NE 68588 USA. NCI, Sect Mol Biol Selenium, Basic Res Lab, NIH, Bethesda, MD 20892 USA. Fidel Syst Inc, Gaithersburg, MD 20879 USA. Univ Illinois, Chicago, IL 60612 USA. Seoul Natl Univ, IMBG, Genet Mol Lab, Seoul 151742, South Korea. Harvard Univ, Brigham & Womens Hosp, Sch Med, Dept Med, Boston, MA 02115 USA. RP Gladyshev, VN (reprint author), Univ Nebraska, Dept Biochem, Beadle Ctr, Lincoln, NE 68588 USA. RI Gladyshev, Vadim/A-9894-2013; Kwon, So Yeon/M-5625-2014 OI Kwon, So Yeon/0000-0002-8490-9101 NR 33 TC 94 Z9 100 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35540 EP 35547 DI 10.1074/jbc.M004014200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800093 PM 10945981 ER PT J AU Wagner, PD Vu, ND AF Wagner, PD Vu, ND TI Phosphorylation of geranyl and farnesyl pyrophosphates by Nm23 proteins/nucleoside diphosphate kinases SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BREAST-CARCINOMA CELLS; ENZYMATIC-ACTIVITY; TUMOR-METASTASIS; MELANOMA-CELLS; PC12 CELLS; MUTATION; GENE; TRANSFECTION; MOTILITY; OVEREXPRESSION AB The biochemical mechanism(s) by which Nm23 proteins/nucleoside diphosphate kinases suppress tumor metastasis, inhibit cell motility, and affect cellular differentiation are not known. Here we report that Nm23 proteins can phosphorylate geranyl and farnesyl pyrophosphates to give triphosphates. Wild type Nm23-H1 had higher geranyl and farnesyl pyrophosphate kinase activities than did mutants of Nm23-H1 that do not inhibit-cell motility. The phosphorylation of farnesyl pyrophosphate appears to occur in vivo as cells with an elevated level of Nm23-H1 contained more farnesyl triphosphate than did control cells. To our knowledge, this is the first report that farnesyl triphosphate exists in cells, The phosphorylation of farnesyl pyrophosphate by Nm23 proteins could alter isoprenoid metabolism, and cells with an elevated level of Nm23 proteins were found to contain more farnesylated 46- and 24-kDa proteins than did control cells. The phosphorylation of geranyl and farnesyl pyrophosphates by Nm23 proteins provides a novel mechanism by which these proteins might-exert their biological effects. C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Wagner, PD (reprint author), NCI, Biochem Lab, NIH, Bldg 37,Rm 4C24, Bethesda, MD 20892 USA. NR 38 TC 20 Z9 22 U1 2 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 10 PY 2000 VL 275 IS 45 BP 35570 EP 35576 DI 10.1074/jbc.M006106200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 375VW UT WOS:000165422800097 PM 10952986 ER PT J AU Cao, XL Tian, Y Zhang, TY Ito, Y AF Cao, XL Tian, Y Zhang, TY Ito, Y TI Supercritical fluid extraction of catechins from Cratoxylum prunifolium Dyer and subsequent purification by high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE counter-current chromatography; Cratoxylum prunifolium; tea; food analysis; catechins ID PRODUCTS AB Supercritical fluid extraction of tea catechins including epigallocatechin-3-O-gallate (EGCG) and epicatechin-3-O-gallate (ECG) from Cratoxylum prunifolium Dyer was performed. The optimization of parameters was carried out using an analytical-scale supercritical fluid extraction (SFE) system designed in our laboratory. Then the extraction was scaled up by 100 times using a preparative SFE system under a set of optimized conditions of 40 degreesC, 25 MPa and modified CO2 with 80% ethanol aqueous solution. The combined yield of EGCG and ECG reached about 1 mg per 1 g of tea leaves where the solubility was near 1.4x10(-4) mass fraction of CO2 fluid. EGCG and ECG of high purity (>98%) were obtained from the crude preparative extract by high-speed counter-current chromatography. (C) 2000 Published by Elsevier Science B.V. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. Beijing Inst New Technol Appl, Beijing 100035, Peoples R China. RP Ito, Y (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 10,Room 7N322,10 Ctr Dr MSC 1676, Bethesda, MD 20892 USA. NR 8 TC 21 Z9 24 U1 1 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD NOV 10 PY 2000 VL 898 IS 1 BP 75 EP 81 DI 10.1016/S0021-9673(00)00788-3 PG 7 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 371VU UT WOS:000165200600007 PM 11185625 ER PT J AU Voloshin, ON Wang, LJ Camerini-Otero, RD AF Voloshin, ON Wang, LJ Camerini-Otero, RD TI The homologous pairing domain of RecA also mediates the allosteric regulation of DNA binding and ATP hydrolysis: A remarkable concentration of functional residues SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE allosteric regulation; ATP hydrolysis; G proteins; RecA; RGS ID SINGLE-STRANDED-DNA; REPLICATION PROTEIN-A; ESCHERICHIA-COLI; GTP-HYDROLYSIS; RECOMBINATION PROTEINS; RAD51 RECOMBINASE; CRYSTAL-STRUCTURE; MEIOTIC CELLS; LOOP L2; YEAST AB Switching between the active (ATP and DNA bound) and inactive conformations of the homologous recombination RecA protein is regulated by Am hydrolysis. First, we use the homologous pairing domain of RecA derived from its mobile loop L2 to show that the interaction of this random coil peptide with the gamma -phosphate of ATP results in a peptide beta -conformation similar to that previously shown to be induced by DNA binding. Next, we show that in the whole RecA protein two residues in this L2 domain, Gln194 and Arg196, are catalytic amino acid residues for ATP hydrolysis and functionally resemble the corresponding residues engaged in GTP hydrolysis by two distinct classes of G proteins. Finally, we show that the role of DNA and high salt in the stimulation of the ATPase of RecA is to stabilize this highly mobile region involved in hydrolysis. This is a role similar to that described for RGSs in the activation of the GTPase of heterotrimeric G proteins. Therefore, (i) a prototypical DNA-dependent ATPase and ATP-stimulated DNA-binding protein, RecA, and eukaryotic signaling proteins share common stereochemical regulatory mechanisms; and (ii) in a remarkable example of parsimony, loop L2 is a molecular switch that controls both ATP promoted DNA binding and pairing reactions and DNA stimulated ATP hydrolysis. (C) 2000 Academic Press. C1 NIDDK, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. RP Camerini-Otero, RD (reprint author), NIDDK, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. NR 63 TC 31 Z9 31 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD NOV 10 PY 2000 VL 303 IS 5 BP 709 EP 720 DI 10.1006/jmbi.2000.4163 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 373QH UT WOS:000165299800006 PM 11061970 ER PT J AU Deng, XL Cadet, JL AF Deng, XL Cadet, JL TI Methamphetamine-induced apoptosis is attenuated in the striata of copper-zinc superoxide dismutase transgenic mice SO MOLECULAR BRAIN RESEARCH LA English DT Article DE methamphetamine; apoptosis; TUNEL; caspase ID POLY(ADP-RIBOSE) POLYMERASE; INDUCED NEUROTOXICITY; P53-KNOCKOUT MICE; CELL-DEATH; DOPAMINE; NEURONS; DAMAGE AB Administration of methamphetamine caused significant increases in terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells, in poly (ADP-ribose) polymerase (PARP) cleavage, as well as in caspase-3 activity in the striata of C57BL/6J mice. In contrast, all these effects were markedly suppressed in the copper-zinc superoxide dismutase transgenic mice. These results indicate that superoxide radicals might be important factors in METH-induced cell death. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDA, Mol Neuropsychiat Sect, Intramurla Res Program, Baltimore, MD 21224 USA. RP Cadet, JL (reprint author), NIDA, Mol Neuropsychiat Sect, Intramurla Res Program, Baltimore, MD 21224 USA. NR 24 TC 49 Z9 50 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD NOV 10 PY 2000 VL 83 IS 1-2 BP 121 EP 124 DI 10.1016/S0169-328X(00)00169-8 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 374JQ UT WOS:000165342400012 ER PT J AU Hou, XH Foley, S Cueto, M Robinson, MA AF Hou, XH Foley, S Cueto, M Robinson, MA TI The human T-cell leukemia virus type I (HTLV-I) X region encoded protein p13(II) interacts with cellular proteins SO VIROLOGY LA English DT Article ID ACTIN-BINDING PROTEIN; CYTOPLASMIC DOMAIN; PX REGION; SEQUENCE; FILAMIN; IDENTIFICATION; LOCOMOTION; EXPRESSION; EFFICIENT; LYMPHOMA AB Interactions between the Human T-cell leukemia virus type I (HTLV-I) gene product p13 " and cellular proteins were investigated using the yeast two-hybrid system. Variant forms of p13 " were derived from two HTLV-I molecular clones, K30p and K34p, that differ in both Virus production and in vivo and in vitro infectivity. Two nucleotide differences between the p13 from K30p (p13K30) and K34p (p13K34) result in a Trp-Arg substitution at amino acid 17 and the truncation of the 25 carboxyl-terminal residues of p13K34. A cDNA library from an HTLV-1-infected rabbit T-cell line was screened with p13K30 and p13K34 as bait, products of two cDNA clones, C44 and C254, interacted with p13K34 but not with p13K30. Interactions were further confirmed using the GST;fusion protein coprecipitation assay. Sequence analysis of C44 and C254 cDNA clones revealed similarities to members of the nucleoside monophosphate kinase superfamily and actin-binding protein 280, respectively. Further analysis of the function of these two proteins and the consequence of their interaction with p13 may help elucidate a role for p13 in virus production, infectivity, or the pathogenesis of HTLV-I. (C) 2000 Academic Press. C1 NIAID, Immunogenet Lab, NIH, Rockville, MD 20852 USA. Novartis Inst Biomed Res Biomol Struct & Comp CTA, Summit, NJ 07901 USA. RP Robinson, MA (reprint author), NIAID, Immunogenet Lab, NIH, Twinbrook 2 Facil,12441 Parklawn Dr, Rockville, MD 20852 USA. NR 41 TC 13 Z9 15 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD NOV 10 PY 2000 VL 277 IS 1 BP 127 EP 135 DI 10.1006/viro.2000.0604 PG 9 WC Virology SC Virology GA 376WX UT WOS:000165480700015 PM 11062043 ER PT J AU Braun, E Rorman, E Lueders, KK Bar-Sinai, A Hochman, J AF Braun, E Rorman, E Lueders, KK Bar-Sinai, A Hochman, J TI Differential expression of intracisternal A-particle transcripts in immunogenic versus tumorigenic S49 murine lymphoma cells SO VIROLOGY LA English DT Article ID LONG TERMINAL REPEAT; MOUSE MAMMARY-TUMOR; RETROVIRAL PARTICLE; PROVIRAL ELEMENTS; GENES; SEQUENCE; METHYLATION; ANTIBODIES; ACTIVATION; PROTEINS AB Tumorigenic S49 mouse lymphoma cells (T-25) were compared to their nontumorigenic (immunogenic) substrate-adherent descendants (T-25-Adh), using the differential display technique. A 784-bp fragment with 92% sequence homology to the intracisternal A-particle (IAP) element family was isolated from the latter cells. IAP sequences are endogenous, noninfectious retroviral elements that can undergo transpositions and act as mutagens. Expression of IAP transcripts (as detected by the isolated fragment) was 5- to 10-fold higher in T-25-Adh cells than in T-25 cells. IAP RT-PCR cDNA clones derived from the immunogenic T-25-Adh cells, but not from T-25 cells, contain two distinctive motifs: (i) a motif characteristic of IAP elements expressed in lymphoid cells (lymphocyte specific, LS); (ii) a nonapeptide sequence known to stimulate cytotoxic T lymphocytes in a leukemia cell line expressing IAP sequences. In addition, expression of transcripts containing these motifs is enhanced in the immunogenic cells as opposed to the tumorigenic cells. Furthermore, one of the IAP elements (belonging to the LSI subfamily) is specifically hypomethylated in the DNA of the immunogenic cells. The above-mentioned relationship was strengthened when tumorigenic revertants derived from T-25-Adh cells, as well as independently selected tumorigenic and immunogenic S49 sublines, were studied. In all cases, enhanced immunogenicity was linked to the up-regulation of specific IAP elements. No transpositions of LS1 elements were observed among the different sublines studied. These findings suggest that, in the S49 lymphoma, selectively expressed IAP retroviral elements may function in a tumor suppressive capacity by affecting the immunogenic potential of these cells. (C) 2000 Academic Press. C1 Hebrew Univ Jerusalem, Dept Cell & Anim Biol, IL-91904 Jerusalem, Israel. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Hochman, J (reprint author), Hebrew Univ Jerusalem, Dept Cell & Anim Biol, IL-91904 Jerusalem, Israel. NR 41 TC 6 Z9 6 U1 0 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD NOV 10 PY 2000 VL 277 IS 1 BP 136 EP 146 DI 10.1006/viro.2000.0568 PG 11 WC Virology SC Virology GA 376WX UT WOS:000165480700016 PM 11062044 ER PT J AU Sosnovtsev, SV Green, KY AF Sosnovtsev, SV Green, KY TI Identification and genomic mapping of the ORF3 and VPg proteins in feline calicivirus virions SO VIROLOGY LA English DT Article ID HEMORRHAGIC-DISEASE VIRUS; CAPSID PRECURSOR PROTEIN; INFECTED-CELLS; NUCLEOTIDE-SEQUENCE; LINKED PROTEIN; RNA-SYNTHESIS; EXPRESSION; TRANSLATION; POLYPEPTIDE; BACULOVIRUS AB Two minor proteins with molecular masses of 8.5 and 15.5 kDa were identified in feline calicivirus (FCV) virions. Direct sequence analysis showed that the N-terminal sequence of the 8.5-kDa protein was identical to that of the predicted protein encoded by open reading frame 3 (ORF3) of the FCV genome. The N-terminal sequence of the 15.5-kDa protein corresponded to amino acids 961-980 of the FCV ORF1 polyprotein and mapped to the genomic location of the calicivirus VPS. Antisera raised against recombinant ORF3 protein or the N-terminal 20 amino acids of the putative VPS reacted with the corresponding proteins present in both a Western blot analysis of purified FCV virions and an immunofluorescence assay of FCV-infected cells. A comparative analysis of radioactivity incorporated into virion proteins during in vivo labeling experiments indicated that the ORF3 protein is likely present in one or two copies per virion. The mobility of the ORF3 protein present in virions was similar to that of the ORF3 protein found in FCV-infected cells or expressed in bacteria. Direct N- and C-terminal sequence analysis of the purified ORF3 protein obtained by expression in bacteria demonstrated the presence of intact, uncleaved termini, suggesting that the observed difference between the calculated and the apparent masses in SDS-PAGE was not due to proteolytic processing of the protein. C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Sosnovtsev, SV (reprint author), NIAID, Infect Dis Lab, NIH, 900 Rockville Pike,Bldg 7,Room 137, Bethesda, MD 20892 USA. NR 36 TC 64 Z9 66 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD NOV 10 PY 2000 VL 277 IS 1 BP 193 EP 203 DI 10.1006/viro.2000.0579 PG 11 WC Virology SC Virology GA 376WX UT WOS:000165480700022 PM 11062050 ER PT J AU Finkel, T Holbrook, NJ AF Finkel, T Holbrook, NJ TI Oxidants, oxidative stress and the biology of ageing SO NATURE LA English DT Review ID AMYOTROPHIC-LATERAL-SCLEROSIS; TRANSGENIC DROSOPHILA-MELANOGASTER; MANGANESE SUPEROXIDE-DISMUTASE; ACTIVATED PROTEIN-KINASE; EPIDERMAL GROWTH-FACTOR; EXTENDED LIFE-SPAN; CAENORHABDITIS-ELEGANS; HYDROGEN-PEROXIDE; CONTROLLED TRIAL; VITAMIN-E AB Living in an oxygenated environment has required the evolution of effective cellular strategies to detect and detoxify metabolites of molecular oxygen known as reactive oxygen species. Here we review evidence that the appropriate and inappropriate production of oxidants, together with the ability of organisms to respond to oxidative stress, is intricately connected to ageing and life span. C1 NHLBI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NIA, Biol Chem Lab, NIH, Baltimore, MD 21224 USA. RP Finkel, T (reprint author), NHLBI, Mol Biol Lab, NIH, Bldg 10-6N-240,10 Ctr Dr, Bethesda, MD 20892 USA. RI Bell, Tiffany/F-4403-2010 NR 106 TC 4287 Z9 4508 U1 139 U2 998 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD NOV 9 PY 2000 VL 408 IS 6809 BP 239 EP 247 DI 10.1038/35041687 PG 9 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 371LL UT WOS:000165180400054 PM 11089981 ER PT J AU Merke, DP Chrousos, GP Eisenhofer, G Weise, M Keil, MF Rogol, AD Van Wyk, JJ Bornstein, SR AF Merke, DP Chrousos, GP Eisenhofer, G Weise, M Keil, MF Rogol, AD Van Wyk, JJ Bornstein, SR TI Adrenomedullary dysplasia and hypofunction in patients with classic 21-hydroxylase deficiency. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID CONGENITAL ADRENAL-HYPERPLASIA; LIQUID-CHROMATOGRAPHY; CATECHOLAMINES; PLASMA; METANEPHRINES; METABOLISM; DISEASE; STRESS AB Background: Glucocorticoids are essential for the normal development and functioning of the adrenal medulla. Whether adrenomedullary structure and function are normal in patients with congenital adrenal hyperplasia is not known. Methods: We measured plasma and urinary catecholamines and plasma metanephrines in 38 children with congenital adrenal hyperplasia due to 21-hydroxylase deficiency (25 children with the salt-wasting form and 13 with the simple virilizing form), 39 age-matched normal subjects, and 20 patients who had undergone bilateral adrenalectomy. Adrenal specimens obtained from three other patients with 21-hydroxylase deficiency who had undergone bilateral adrenalectomy and specimens obtained at autopsy from eight other patients were examined histologically. Results: Plasma epinephrine and metanephrine concentrations and urinary epinephrine excretion were 40 to 80 percent lower in the patients with congenital adrenal hyperplasia than in the normal subjects (P<0.05), and the values were lowest in the patients with the most severe deficits in cortisol production. Urinary epinephrine excretion and plasma epinephrine concentrations were at or below the limit of detection of the assay in 8 (21 percent) of the patients with congenital adrenal hyperplasia and in 19 (95 percent) of the patients who had undergone adrenalectomy. In the group of patients with congenital adrenal hyperplasia, plasma epinephrine and metanephrine concentrations and urinary epinephrine excretion were approximately 50 percent lower in those who had been hospitalized for adrenal crises than in those who had not. In three patients with congenital adrenal hyperplasia who had undergone bilateral adrenalectomy, the formation of the adrenal medulla was incomplete, and electron-microscopical studies revealed a depletion of secretory vesicles in chromaffin cells. Conclusions: Congenital adrenal hyperplasia compromises both the development and the functioning of the adrenomedullary system. (N Engl J Med 2000;343:1362-8.) (C) 2000, Massachusetts Medical Society. C1 NIH, Warren Grant Magnuson Clin Ctr, Bethesda, MD 20892 USA. NICHHD, Pediat & Reprod Endocrinol Branch, Bethesda, MD 20892 USA. NINDS, Clin Neurocardiol Sect, Bethesda, MD 20892 USA. Univ Virginia Hlth Syst, Charlottesville, VA USA. Univ N Carolina, Sch Med, Dept Pediat, Chapel Hill, NC USA. RP Merke, DP (reprint author), NIH, Warren Grant Magnuson Clin Ctr, Bldg 10,Rm 13S260,10 Ctr Dr,MSC 1932, Bethesda, MD 20892 USA. NR 26 TC 116 Z9 119 U1 0 U2 2 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 NOV 9 PY 2000 VL 343 IS 19 BP 1362 EP 1368 DI 10.1056/NEJM200011093431903 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 370ZH UT WOS:000165152800003 PM 11070100 ER PT J AU Weinstein, JN AF Weinstein, JN TI Pharmacogenomics - Teaching old drugs new tricks SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material ID CPG ISLANDS; CANCER C1 NCI, Bethesda, MD 20892 USA. RP Weinstein, JN (reprint author), NCI, Bethesda, MD 20892 USA. NR 7 TC 43 Z9 45 U1 0 U2 1 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 NOV 9 PY 2000 VL 343 IS 19 BP 1408 EP 1409 DI 10.1056/NEJM200011093431910 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 370ZH UT WOS:000165152800010 PM 11070107 ER PT J AU Johnson, JG Cohen, P Pine, DS Klein, DF Kasen, S Brook, JS AF Johnson, JG Cohen, P Pine, DS Klein, DF Kasen, S Brook, JS TI Association between cigarette smoking and anxiety disorders during adolescence and early adulthood SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID PANIC DISORDER; PSYCHIATRIC-DISORDERS; MAJOR DEPRESSION; SUBSTANCE USE; NICOTINE; ATTACKS AB Context Cigarette smoking is associated with some anxiety disorders, but the direction of the association between smoking and specific anxiety disorders has not been determined. Objective To investigate the longitudinal association between cigarette smoking and anxiety disorders among adolescents and young adults. Design The Children in the Community Study, a prospective longitudinal investigation. Setting and Participants Community-based sample of 688 youths (51% female) from upstate New York interviewed in the years 1985-1986, at a mean age of 16 years, and in the years 1991-1993, at a mean age of 22 years. Main Outcome Measure Participant cigarette smoking and psychiatric disorders in adolescence and early adulthood, measured by age-appropriate versions of the Diagnostic interview Schedule for Children. Results Heavy cigarette smoking (greater than or equal to 20 cigarettes/d) during adolescence was associated with higher risk of agoraphobia (10.3% vs 1.8%; odds ratio [OR], 6.79; 95% confidence interval [CI], 1.53-30.17), generalized anxiety disorder (20.5% vs 3.71%; OR, 5.53; 95% CI, 1.84-16.66), and panic disorder (7.7% vs 0.6%; OR, 15.58; 95% CI, 2.31-105.14) during early adulthood after controlling for age, sex, difficult childhood temperament; alcohol and drug use, anxiety, and depressive disorders during adolescence; and parental smoking, educational level, and psychopathology. Anxiety disorders during adolescence were not significantly associated with chronic cigarette smoking during early adulthood. Fourteen percent and 15% of participants with and without anxiety during adolescence, respectively, smoked at least 20 cigarettes per day during early adulthood (OR, 0.88; 95% CI, 0.36-2.14). Conclusion Our results suggest that cigarette smoking may increase risk of certain anxiety disorders during late adolescence and early adulthood. C1 New York State Psychiat Inst, Dept Psychiat, New York, NY 10032 USA. Columbia Univ, Dept Psychiat, New York, NY USA. Mt Sinai Med Ctr, Dept Community Med, New York, NY 10029 USA. NIMH, Intramural Res Program, Program Mood & Anxiety Disorders, Bethesda, MD 20892 USA. RP Johnson, JG (reprint author), New York State Psychiat Inst, Dept Psychiat, Box 60,1051 Riverside Dr, New York, NY 10032 USA. FU NIDA NIH HHS [DA-03188]; NIMH NIH HHS [K-20-MH-01391, MH-36971] NR 22 TC 279 Z9 288 U1 3 U2 21 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 NOV 8 PY 2000 VL 284 IS 18 BP 2348 EP 2351 DI 10.1001/jama.284.18.2348 PG 4 WC Medicine, General & Internal SC General & Internal Medicine GA 369NU UT WOS:000165074500029 PM 11066185 ER PT J AU El-Gabalawy, HS Duray, P Goldbach-Mansky, R AF El-Gabalawy, HS Duray, P Goldbach-Mansky, R TI Evaluating patients with arthritis of recent onset - Studies in pathogenesis and prognosis SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID EARLY RHEUMATOID-ARTHRITIS; REACTIVE ARTHRITIS; INFLAMMATORY ARTHRITIS; COMBINATION THERAPY; HLA-DRB1 ALLELES; DOUBLE-BLIND; CLASSIFICATION; ASSOCIATION; PROGRESSION; SYNOVITIS AB Inflammatory synovitis of recent onset poses a diagnostic and prognostic challenge to primary care physicians and rheumatologists, A lack of understanding of the underlying etiologic and pathogenic processes limits the ability to distinguish forms of arthritis that follow a benign, self-limiting course from forms that proceed to an aggressive, erosive disease requiring intensive immunosuppressive therapy. It is estimated that between 30% and 40% of patients presenting with early synovitis have disease that remains unclassified. Using data from a cohort of patients with early synovitis and reviewing current literature, we discuss investigational approaches toward a new classification of patients with early synovitis. Although a lack of understanding of this heterogeneous clinical syndrome has led clinicians to take a largely empirical approach to treatment thus far, the evolving awareness of disease predisposition at a genetic level and the expanding ability to specifically manipulate biological pathways may ultimately change the approach to this clinical problem. C1 NIH, Bethesda, MD 20892 USA. RP El-Gabalawy, HS (reprint author), NIH, Bldg 10,Room 9S205, Bethesda, MD 20892 USA. NR 32 TC 13 Z9 13 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 NOV 8 PY 2000 VL 284 IS 18 BP 2368 EP 2373 DI 10.1001/jama.284.18.2368 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 369NU UT WOS:000165074500032 PM 11066188 ER PT J AU Bezrukov, SM Berezhkovskii, AM Pustovoit, MA Szabo, A AF Bezrukov, SM Berezhkovskii, AM Pustovoit, MA Szabo, A TI Particle number fluctuations in a membrane channel SO JOURNAL OF CHEMICAL PHYSICS LA English DT Article ID MALTOPORIN AB Channel-facilitated transport of metabolites across biological membranes results in excess noise in the current carried by small ions. This noise originates from fluctuations of the number of metabolite molecules in the channel due to their diffusion. We have carried out a theoretical study of particle number fluctuations in a cylindrical pore. First, we obtain the power spectral density of these fluctuations as a function of pore length and radius, as well as the diffusion constants of the particle in the pore and in the bulk, in the absence of particle-pore interactions. We then perform three-dimensional Brownian dynamics simulations that show excellent agreement with the analytical result. Finally, we demonstrate that explicit expressions for the low-frequency limit of the spectral density can be found even when the particle interacts with the pore. (C) 2000 American Institute of Physics. [S0021-9606(00)70842-7]. C1 NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. LY Karpov Phys Chem Res Inst, Moscow 103064, Russia. NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. EM bezrukov@helix.nih.gov RI Pustovoit, Mark/B-5249-2008; Szabo, Attila/H-3867-2012 NR 21 TC 70 Z9 70 U1 1 U2 6 PU AMER INST PHYSICS PI MELVILLE PA 1305 WALT WHITMAN RD, STE 300, MELVILLE, NY 11747-4501 USA SN 0021-9606 EI 1089-7690 J9 J CHEM PHYS JI J. Chem. Phys. PD NOV 8 PY 2000 VL 113 IS 18 BP 8206 EP 8211 AR PII [S0021-9606(00)70842-7] DI 10.1063/1.1314862 PG 6 WC Chemistry, Physical; Physics, Atomic, Molecular & Chemical SC Chemistry; Physics GA 369BF UT WOS:000090151400061 ER PT J AU Razzaque, A Dye, E Puri, RK AF Razzaque, A Dye, E Puri, RK TI Characterization of tumor vaccines during product development SO VACCINE LA English DT Article; Proceedings Paper CT 2nd Annual Conference on Vaccine Research CY MAR 27-30, 1999 CL BETHESDA, MARYLAND DE tumor vaccines; product development; regulatory issues ID DENDRITIC CELLS; MALIGNANT GLIOMAS; IN-VITRO; IMMUNOTHERAPY; ANTIGEN; IMMUNIZATION; PEPTIDES; IMMUNITY; VECTOR; RNA AB The term tumor vaccines encompasses a wide variety of diverse agents capable of interacting with the imune system to product local inflammation. delayed type hypersensitivity reaction and/or tumor regression and, hopefully, a therapeutic effect. These vaccines may be grouped into the following general areas: (1) Cell-based vaccines such as manipulated tumor cells, activated peripheral blood or bone marrow-derived lymphocytes, dendritic cells or other antigen presenting cells (APC) and gene-modified tumor cells or other cells engineered to express cytokines, growth factors or tumor antigens. (2) Antigen preparations, such as synthetic peptides. purified antigens and tumor cell lysates. (3) Viral and plasmid vectors expressing therapeutic genes. (4) Liposome containing antigen, peptides, plasmids encoding tumor antigens. While no tumor vaccine has been licensed by the FDA, numerous clinical trials are ongoing and some products have advanced to Phase III pivotal stages of development. However, as with many novel products, major regulatory and scientific issues associated with clinical use of tumor vaccines remain to be addressed. In this paper, we address issues associated with different types of tumor Vaccines and provide recommendations for the characterization of these vaccines at various stages of development. Published by Elsevier Science Ltd. C1 US FDA, Ctr Biol Evaluat & Res, Div Cellular & Gene Therapies, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Div Cytokine Biol, Bethesda, MD 20892 USA. RP Razzaque, A (reprint author), US FDA, Ctr Biol Evaluat & Res, Div Cellular & Gene Therapies, HFM-530,NIH Bldg 29B,Room 2NN10, Bethesda, MD 20892 USA. NR 26 TC 6 Z9 7 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 NOV 8 PY 2000 VL 19 IS 6 BP 644 EP 647 DI 10.1016/S0264-410X(00)00251-6 PG 4 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 375LL UT WOS:000165400800008 PM 11090716 ER PT J AU Felson, DT Lawrence, RC Hochberg, MC McAlindon, T Dieppe, PA Minor, MA Blair, SN Berman, BM Fries, JF Weinberger, M Lorig, KR Jacobs, JJ Goldberg, V AF Felson, DT Lawrence, RC Hochberg, MC McAlindon, T Dieppe, PA Minor, MA Blair, SN Berman, BM Fries, JF Weinberger, M Lorig, KR Jacobs, JJ Goldberg, V TI Osteoarthritis: New insights - Part 2: Treatment approaches SO ANNALS OF INTERNAL MEDICINE LA English DT Editorial Material ID NONSTEROIDAL ANTIINFLAMMATORY DRUGS; SERIOUS GASTROINTESTINAL COMPLICATIONS; RANDOMIZED CONTROLLED TRIAL; TOTAL HIP-ARTHROPLASTY; HEALTH-CARE COSTS; RHEUMATOID-ARTHRITIS; KNEE OSTEOARTHRITIS; ARTICULAR-CARTILAGE; PEPTIC-ULCER; DOUBLE-BLIND AB Osteoarthritis is the most common form of arthritis, affecting millions of people in the United States. It is a complex disease whose etiology bridges biomechanics and biochemistry. Evidence is growing for the role of systemic factors, such as genetics, diet, estrogen use, and bone density, and local biomechanical factors, such as muscle weakness, obesity, and joint laxity. These risk factors are particularly important in the weight-bearing joints, and modifying them may help prevent osteoarthritis-related pain and disability. Major advances in management to reduce pain and disability are yielding a panoply of available treatments ranging from nutriceuticals to chondrocyte transplantation, new oral antiinflammatory medications, and health education. This article is part 2 of a two-part summary of a National Institutes of Health conference that brought together experts in osteoarthritis from diverse backgrounds and provided a multidisciplinary and comprehensive summary of recent advances in the prevention of osteoarthritis onset, progression, and disability. Part 2 focuses on treatment approaches; evidence for the efficacy of commonly used oral therapies is reviewed and information on alternative therapies, including nutriceuticals and acupuncture, is presented. Biomechanical interventions, such as exercise and bracing, and behavioral interventions directed toward enhancing self-management are reviewed. Current surgical approaches are described and probable future biotechnology-oriented approaches to treatment are suggested. C1 NIAMSD, NIH, Bethesda, MD 20892 USA. Boston Univ, Sch Med, Boston, MA 02118 USA. Univ Maryland, Sch Med, Baltimore, MD 21201 USA. Univ Bristol, MRC Hlth Serv Res Collaborat, Bristol, Avon, England. Univ Missouri, Columbia, MO USA. Cooper Inst Aerob Res, Dallas, TX USA. Stanford Univ, Sch Med, Palo Alto, CA 94304 USA. Indiana Univ, Indianapolis, IN 46204 USA. Rush Presbyterian St Lukes Med Ctr, Chicago, IL 60612 USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. RP Lawrence, RC (reprint author), NIAMSD, NIH, Bldg 45 Room 5AS-37G, Bethesda, MD 20892 USA. RI Fries, James/F-6271-2011 NR 135 TC 270 Z9 280 U1 6 U2 59 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 J9 ANN INTERN MED JI Ann. Intern. Med. PD NOV 7 PY 2000 VL 133 IS 9 BP 726 EP 737 PG 12 WC Medicine, General & Internal SC General & Internal Medicine GA 369PC UT WOS:000165075300008 PM 11074906 ER PT J AU Sayers, EW Gerstner, RB Draper, DE Torchia, DA AF Sayers, EW Gerstner, RB Draper, DE Torchia, DA TI Structural preordering in the N-terminal region of ribosomal protein S4 revealed by heteronuclear NMR spectroscopy SO BIOCHEMISTRY LA English DT Article ID TRIPLE-RESONANCE NMR; SIDE-CHAIN RESONANCES; C-13/N-15-ENRICHED PROTEINS; ESCHERICHIA-COLI; C-13-ENRICHED PROTEINS; C-13 MAGNETIZATION; COUPLING-CONSTANTS; RNA RECOGNITION; LARGER PROTEINS; BACKBONE AMIDE AB Protein S4, a component of the 30S subunit of the prokaryotic ribosome, is one of the first proteins to interact with rRNA in the process of ribosome assembly and is known to be involved in the regulation of this process. While the structure of the C-terminal 158 residues of Bacillus stearothermophilus S4 has been solved by both X-ray crystallography and NMR, that of the N-terminal 41 residues is unknown. Evidence suggests that the N-terminus is necessary both for the assembly of functional ribosomes and for full binding to 16S RNA, and so we present NMR data collected on the full-length protein (200 aa), Our data indicate that the addition of the N-terminal residues does not significantly change the structure of the C-terminal 158 residues. The data further indicate that the N-terminus is highly flexible in solution, without discernible secondary structure. Nevertheless, structure calculations based on nuclear Overhauser effect spectroscopic data combined with N-15 relaxation data revealed that two short segments in the N-terminus, S12RRL15 and P30YPP33, adopt transiently ordered states in solution. The major conformation of S12RRL15 appears to orient the arginine side chains outward toward the solvent in a parallel fashion, while that of P30YPP33 forms a nascent turn of a polyproline II helix. These segments contain residues that are highly conserved across many prokaryotic species, and thus they are reasonable candidates respectively for sites of interaction with RNA and other ribosomal proteins within the intact ribosome. C1 Natl Inst Dent & Craniofacial Res, NIH, Struct Mol Biol Unit, Bethesda, MD 20892 USA. Johns Hopkins Univ, Dept Chem, Baltimore, MD 21218 USA. RP Torchia, DA (reprint author), Natl Inst Dent & Craniofacial Res, NIH, Struct Mol Biol Unit, 30 Convent Dr,MSC 4307, Bethesda, MD 20892 USA. NR 59 TC 43 Z9 45 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD NOV 7 PY 2000 VL 39 IS 44 BP 13602 EP 13613 DI 10.1021/bi0013391 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 372BR UT WOS:000165214200034 PM 11063598 ER PT J AU Self, WT Tsai, L Stadtman, TC AF Self, WT Tsai, L Stadtman, TC TI Synthesis and characterization of selenotrisulfide-derivatives of lipoic acid and lipoamide SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID MAMMALIAN THIOREDOXIN REDUCTASE; ESCHERICHIA-COLI; SELENIUM METABOLISM; DEHYDROGENASE; PROTEIN; MECHANISM; ANALOG AB Thiol-containing compounds, such as glutathione and cysteine. react with selenite under specific conditions to form selenotrisulfides. Previous studies have focused on isolation and characterization of intermolecular selenotrisulfides. This study describes the preparation and characterization of intramolecular selenotrisulfide derivatives of lipoic acid and lipoamide. These derivatives, after separation from other reaction products by reverse-phase HPLC, exhibit an absorbance maximum at 288 nm with an extinction coefficient of 1.500 M-1.cm(-1). The selenotrisulfide derivative of lipoic acid was significantly stable at or below pH 8.0 in contrast to several other previously studied selenotrisulfides. Mass spectral analysis of the lipoic acid and lipoamide derivatives confirmed both the expected molecular weights and also the presence of a single atom of selenium as revealed by its isotopic distribution. The selenotrisulfide derivative of lipoic acid was found to serve as an effective substrate for recombinant human thioredoxin reductase as well as native rat thioredoxin reductase in the presence of NADPH. Likewise, the lipoamide derivative was efficiently reduced by NADH-dependent bovine lipoamide dehydrogenase. The significant in vitro stability of these intramolecular selenotrisulfide derivatives of lipoic acid can serve as an important asset in the study of such selenium adducts as model selenium donor compounds for selenophosphate biosynthesis and as rate enhancement effecters in various redox reactions. C1 NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Self, WT (reprint author), NHLBI, Biochem Lab, NIH, Bldg 3,Room 111,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Self, William/A-6704-2008 NR 30 TC 27 Z9 30 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 NOV 7 PY 2000 VL 97 IS 23 BP 12481 EP 12486 DI 10.1073/pnas.220426897 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800026 PM 11050172 ER PT J AU Tikkanen, MK Carter, DJ Harris, AM Le, HM Azorsa, DO Meltzer, PS Murdoch, FE AF Tikkanen, MK Carter, DJ Harris, AM Le, HM Azorsa, DO Meltzer, PS Murdoch, FE TI Endogenously expressed estrogen receptor and coactivator AIB1 interact in MCF-7 human breast cancer cells SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID GENE AB Coactivators are believed to mediate estrogen-induced gene responses via interaction with estrogen receptors (ER), Currently, a major challenge is to determine the importance of each coactivator in a specific cell type and promoter context in response to a particular ligand, The potential of ER to interact with a growing list of coactivators has been shown in a variety of in vitro and gene transfer assays, yet very few data have demonstrated the interaction of endogenous coactivators with ER in intact cells. We report here a ligand-specific interaction of endogenous human ER (hER) and the AIB1 coactivator in MCF-7 human breast cancer cells by using immunoprecipitation analyses. Complexes between endogenously expressed hER and AIB1 were detected in estradiol-treated cells and to a much lesser extent in cells treated with the partial agonist, monohydroxytamoxifen. We were unable to detect an hER-SRC-1 complex in our immunoprecipitations from MCF-7 cells. The in vitro-binding affinity for mouse ER interaction with AIB1 was estimated to be 40-120 nM. We conclude that AIB1 is a major coactivator for hER in MCF-7 human breast cancer cells. C1 Univ Wisconsin, Dept Pathol & Lab Med, Madison, WI 53706 USA. Hosp Loimaa, Loimaa 32201, Finland. Human Genome Sci Inc, Rockville, MD 20850 USA. US Patent Off, Arlington, VA 22204 USA. Univ Virginia, Sch Med, Charlottesville, VA 22906 USA. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. RP Murdoch, FE (reprint author), Univ Wisconsin, Dept Pathol & Lab Med, MSC 5330,1300 Univ Ave, Madison, WI 53706 USA. FU NIDDK NIH HHS [DK50028] NR 24 TC 35 Z9 35 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 NOV 7 PY 2000 VL 97 IS 23 BP 12536 EP 12540 DI 10.1073/pnas.220427297 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800036 PM 11050174 ER PT J AU Liu, X Shu, S Yamashita, RA Xu, YH Korn, ED AF Liu, X Shu, S Yamashita, RA Xu, YH Korn, ED TI Chimeras of Dictyostelium myosin II head and neck domains with Acanthamoeba or chicken smooth muscle myosin II tail domain have greatly increased and unregulated actin-dependent MgATPase activity SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ACTIVATED ATPASE ACTIVITY; 3 PHOSPHORYLATION SITES; LIGHT-CHAIN KINASE; MOTOR DOMAIN; FILAMENTS; BINDING; ROD; SEGMENT; END AB Phosphorylation of the regulatory light chain of Dictyostelium myosin II increases V-max of its actin-dependent MgATPase activity about 5-fold under normal assay conditions. Under these assay conditions, unphosphorylated chimeric myosins in which the tail domain of the Dictyostelium myosin II heavy chain is replaced by either the tail domain of chicken gizzard smooth muscle or Acanthamoeba myosin II are 20 times more active because of a 10- to 15-fold increase in V-max and a 2- to 7-fold decrease in apparent K-ATPase and are only slightly activated by regulatory light chain phosphorylation. Actin-dependent MgATPase activity of the Dictyostelium/Acanthamoeba chimera is not affected by phosphorylation of serine residues in the tail whose phosphorylation completely inactivates wild-type Acanthamoeba myosin II. These results indicate that the actin-dependent MgATPase activity of these myosins involves specific, tightly coupled, interactions between head and tail domains. C1 NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Dept Mol Cardiol, NIH, Bethesda, MD 20892 USA. RP NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. EM edk@nih.gov RI Korn, Edward/F-9929-2012 NR 38 TC 5 Z9 5 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 NOV 7 PY 2000 VL 97 IS 23 BP 12553 EP 12558 DI 10.1073/pnas.230441497 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800039 PM 11058169 ER PT J AU Korn, ED AF Korn, ED TI Coevolution of head, neck, and tail domains of myosin heavy chains SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID UNCONVENTIONAL MYOSIN; FILAMENT FORMATION; SEQUENCE AB Myosins, a large family of actin-based motors, have one or two heavy chains with one or more light chains associated with each heavy chain. The heavy chains have a (generally) N-terminal head domain with an ATPase and actin-binding site, followed by a neck domain to which the light chains bind, and a C-terminal tail domain through which the heavy chains self-associate and/or bind the myosin to its cargo. Approximately 140 members of the myosin superfamily have been grouped into 17 classes based on the sequences of their head domains. I now show that a phylogenetic tree based on the sequences of the combined neck and tail domains groups 144 myosins, with a few exceptions, into the same 17 classes. For the nine myosin classes that have multiple members, phylogenetic trees based on the head domain or the combined neck/tail domains are either identical or very similar. For class II myosins. very similar phylogenetic: trees are obtained for the head, neck, and tail domains of 47 heavy chains and for 29 essential light chains and 19 regulatory light chains. These data strongly suggest that the head, neck, and tail domains of all myosin heavy chains. and light chains at least of class II myosins, have coevolved and are likely to be functionally interdependent, consistent with biochemical evidence showing that regulated actin-dependent MgATPase activity of Dictyostelium myosin II requires isoform specific interactions between the heavy chain head and tail and light chains. C1 NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Korn, ED (reprint author), NHLBI, Cell Biol Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. RI Korn, Edward/F-9929-2012 NR 13 TC 52 Z9 52 U1 0 U2 5 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD NOV 7 PY 2000 VL 97 IS 23 BP 12559 EP 12564 DI 10.1073/pnas.230441597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800040 PM 11058170 ER PT J AU Sun, YN Kolligs, FT Hottiger, MO Mosavin, R Fearon, ER Nabel, GJ AF Sun, YN Kolligs, FT Hottiger, MO Mosavin, R Fearon, ER Nabel, GJ TI Regulation of beta-catenin transformation by the p300 transcriptional coactivator SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DEPENDENT KINASES; CELL-LINES; CBP; PROTEIN; E1A; ACTIVATION; TCF; ONCOPROTEIN; CARCINOMA; ADAPTER AB The beta -catenin protein plays a critical role in embryonic development and mature tissue homeostasis through its effects on E-cadherin-mediated cell adhesion and Wnt-dependent signal transduction. In colon and other cancers, mutations of beta -catenin or the adenomatous polyposis coli (APC) tumor suppressor appear to stabilize beta -catenin and enhance its interaction with T cell factor (TCF) or lymphoid enhancer factor (Lef) transcription factors. At present, a complete picture of the means by which beta -catenin's interactions with TCF/Lef proteins contribute to neoplastic transformation is lacking. We report that the transcriptional coactivator p300 interacts with beta -catenin in vitro and in vivo and is critical for beta -catenin-mediated neoplastic transformation. p300 synergistically activates beta -catenin/TCF transcription, and their biochemical association requires the CH1 domain of p300 and a region of beta -catenin that includes its NH2-terminal transactivation domain and the first two armadillo repeats. Lowering of cellular p300 levels by using a ribozyme directed against p300 reduced TCF transcriptional activity and inhibited the neoplastic growth properties of a beta -catenin-transformed rat epithelial cell line and a human colon carcinoma line with a beta -catenin mutation. These findings demonstrate a critical role for p300 in beta -catenin/TCF transcription and in cancers arising from defects in beta -catenin regulation. C1 NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. Univ Michigan, Sch Med, Dept Internal Med, Div Med Genet, Ann Arbor, MI 48109 USA. Univ Michigan, Sch Med, Dept Human Genet & Pathol, Ann Arbor, MI 48109 USA. RP Nabel, GJ (reprint author), NIH, Vaccine Res Ctr, 40 Convent Dr, Bethesda, MD 20892 USA. RI Hottiger, Michael/J-7747-2013 OI Hottiger, Michael/0000-0002-7323-2270 NR 33 TC 86 Z9 90 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD NOV 7 PY 2000 VL 97 IS 23 BP 12613 EP 12618 DI 10.1073/pnas.220158597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800049 PM 11050151 ER PT J AU Meng, QX Walker, DM Olivero, OA Shi, XC Antiochos, BB Poirier, MC Walker, VE AF Meng, QX Walker, DM Olivero, OA Shi, XC Antiochos, BB Poirier, MC Walker, VE TI Zidovudine-didanosine coexposure potentiates DNA incorporation of zidovudine and mutagenesis in human cells SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HIV-1 REVERSE-TRANSCRIPTASE; PLACEBO-CONTROLLED TRIAL; AIDS-RELATED COMPLEX; NUCLEOSIDE ANALOGS; AZIDOTHYMIDINE AZT; DRUG-RESISTANCE; DOUBLE-BLIND; 3'-AZIDO-3'-DEOXYTHYMIDINE; MUTATIONS; TOXICITY AB Drug combinations that include nucleoside reverse transcriptase inhibitors (NRTIs) are remarkably effective in preventing maternal-viral transmission of HIV during pregnancy. However, there may be potential long-term risks for children exposed in utero. Examination of the genotoxic and mutagenic effects of two NRTIs, zidovudine [AZT (3'-azido-3'-deoxythymidine)] and didanosine [ddl (2',3'-dideoxyinosine)]. in cultured human lymphoblastoid cells revealed multiplicative synergistic enhancement of AZT-DNA incorporation and mutant frequency induction in response to the combined drug exposure, as compared with single-drug exposures. Dose-related increases in DNA incorporation of AZT (as measured by a competitive RIA) and mutagenicity at the HPRT and TK loci (as assessed by cell-cloning assays) were observed in cells exposed in culture to AZT, or equimolar combinations of AZT + ddl, at exposure concentrations ranging from 3 to 30 times the maximum plasma levels found in humans. Because mutagenesis is strongly associated with tumor induction in experimental models, children exposed transplacentally to combinations of NRTIs may be at risk for cancer development later in life. C1 New York State Dept Hlth, Wadsworth Ctr, Lab Human Toxicol & Mol Epidemiol, Albany, NY 12201 USA. Expt Pathol Labs Inc, Res Triangle Pk, NC 27713 USA. NCI, Cellular Carcinogenesis & Tumor Promot Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. SUNY Albany, Sch Publ Hlth, Dept Environm Hlth & Toxicol, Albany, NY 12203 USA. RP Walker, VE (reprint author), Lovelace Resp Res Inst, Inhalat Toxicol Lab, POB 5890, Albuquerque, NM 87185 USA. FU NICHD NIH HHS [R01HD33648] NR 29 TC 51 Z9 52 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 NOV 7 PY 2000 VL 97 IS 23 BP 12667 EP 12671 DI 10.1073/pnas.220203197 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800058 PM 11058153 ER PT J AU Maliszewska-Tkaczyk, M Jonczyk, P Bialoskorska, M Schaaper, RM Fijalkowska, IJ AF Maliszewska-Tkaczyk, M Jonczyk, P Bialoskorska, M Schaaper, RM Fijalkowska, IJ TI SOS mutator activity: Unequal mutagenesis on leading and lagging strands SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ESCHERICHIA-COLI CHROMOSOME; DNA-REPLICATION; RECA PROTEIN; DINB GENE; POLYMERASE; MODULATION; MUTATIONS; DELETION; MUTANTS; LAMBDA AB A major pathway of mutagenesis in Escherichia coli is mediated by the inducible SOS response. Current models of SOS mutagenesis invoke the interaction of RecA and UmuD'C-2 proteins with a stalled DNA replication complex at sites of DNA lesions or poorly extendable terminal mismatches, resulting in an (error-prone) continuation of DNA synthesis. The precise mechanisms of SOS-mediated lesion bypass or mismatch extension are not known. Here, we have studied mutagenesis on the E. coli chromosome in recA730 strains. In recA730 strains, the SOS system is expressed constitutively. resulting in a spontaneous mutator effect (SOS mutator) because of reduced replication fidelity. We investigated whether during SOS mutator activity replication fidelity might be altered differentially in the leading and lagging strand of replication. Pairs of recA730 strains were constructed differing in the orientation of the lac operon relative to the origin of replication. The strains were also mismatch-repair defective (mutL) to facilitate scoring of replication errors. Within each pair, a given lac sequence is replicated by the leading-strand machinery in one orientation and by the lagging-strand machinery in the other orientation. Measurements of defined lac mutant frequencies in such pairs revealed large differences between the two orientations. Furthermore, in all cases, the frequency bias was the opposite of that seen in normal cells. We suggest that, for the lacZ target used in this study, SOS mutator activity operates with very different efficiency in the two strands. Specifically, the lagging strand of replication appears most susceptible to the SOS mutator effect. C1 Polish Acad Sci, Inst Biochem & Biophys, PL-02106 Warsaw, Poland. NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. RP Fijalkowska, IJ (reprint author), Polish Acad Sci, Inst Biochem & Biophys, Pawinskiego 5A, PL-02106 Warsaw, Poland. RI Fijalkowska, Iwona/I-7796-2016 NR 39 TC 39 Z9 39 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 NOV 7 PY 2000 VL 97 IS 23 BP 12678 EP 12683 DI 10.1073/pnas.220424697 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800060 PM 11050167 ER PT J AU Radu, DL Noben-Trauth, N Hu-Li, J Paul, WE Bona, CA AF Radu, DL Noben-Trauth, N Hu-Li, J Paul, WE Bona, CA TI A targeted mutation in the IL-4R alpha gene protects mice against autoimmune diabetes SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NOD MICE; INTERFERON-GAMMA; INTERLEUKIN-4-DEFICIENT MICE; T-CELLS; TRANSGENIC MODEL; VIRUS-INFECTION; IL-4 RECEPTOR; BETA-CELLS; TH2 CELLS; INSULIN AB Autoimmune insulin-dependent diabetes mellitus (IDDM) occurs spontaneously in mice-bearing transgenes encoding the influenza hemagglutinin under the control of the rat insulin promoter and a T cell receptor specific for an hemagglutinin peptide associated with I-E-d. Such "double transgenic" mice expressing wild-type or targeted IL-4R alpha genes were examined for the onset of IDDM. Eight of 11 mice homozygous for wild-type IL-4R alpha were hyperglycemic by 8 weeks of age, whereas only 1 of 16 mice homozygous for the targeted allele were hyperglycemic at this time. Most IL-4R alpha-/- mice remained normoglycemic to 36 weeks of age. Although only 10% of double transgenic mice homozygous for the wild-type IL-4R alpha allele survived to 30 weeks, 80% of mice homozygous for the targeted allele did so. Heterozygous mice displayed an intermediate frequency of diabetes. Even as late as 270 days of age, mice homozygous for the targeted allele had no insulitis or only peri-insulitis. Thus, the inability to respond to IL-4 and/or IL-13 protects mice against IDDM in this model of autoimmunity. C1 Mt Sinai Sch Med, Dept Microbiol, New York, NY 10029 USA. NIAID, Immunol Lab, Bethesda, MD 20892 USA. RP Bona, CA (reprint author), Mt Sinai Sch Med, Dept Microbiol, New York, NY 10029 USA. FU NIAID NIH HHS [P01 AI024671, P0IAI24671] NR 36 TC 21 Z9 21 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD NOV 7 PY 2000 VL 97 IS 23 BP 12700 EP 12704 DI 10.1073/pnas.230431397 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800064 PM 11050183 ER PT J AU Hussain, SP Raja, K Amstad, PA Sawyer, M Trudel, LJ Wogan, GN Hofseth, LJ Shields, PG Billiar, TR Trautwein, C Hohler, T Galle, PR Phillips, DH Markin, R Marrogi, AJ Harris, CC AF Hussain, SP Raja, K Amstad, PA Sawyer, M Trudel, LJ Wogan, GN Hofseth, LJ Shields, PG Billiar, TR Trautwein, C Hohler, T Galle, PR Phillips, DH Markin, R Marrogi, AJ Harris, CC TI Increased p53 mutation load in nontumorous human liver of Wilson disease and hemochromatosis: Oxyradical overload diseases SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE Wilson disease; hemochromatois; p53; iron; copper; liver carcinogenesis ID TUMOR-SUPPRESSOR GENE; DNA-DAMAGE; NITRIC-OXIDE; HEREDITARY HEMOCHROMATOSIS; LIPID-PEROXIDATION; IN-VIVO; HEPATOCELLULAR-CARCINOMA; HYDROGEN-PEROXIDE; STRAND BREAKS; COPPER AB Hemochromatosis and Wilson disease (WD), characterized by the excess hepatic deposition of iron and copper, respectively, produce oxidative stress and increase the risk of liver cancer. Because the frequency of p53 mutated alleles in nontumorous human tissue may be a biomarker of oxyradical damage and identify individuals at increased cancer risk, we have determined the frequency of p53 mutated alleles in nontumorous liver tissue from WD and hemochromatosis patients. When compared with the liver samples from normal controls, higher frequencies of G:C to T:A transversions at codon 249 (P < 0.001) and C:G to A:T transversions and C:C to T:A transitions at codon 250 (P < 0.001 and P < 0.005) were found in liver tissue from WD cases, and a higher frequency of G:C to T:A transversions at codon 249 (P < 0.05) also was found in liver tissue from hemochromatosis cases. Sixty percent of the WD and 28% of hemochromatosis cases also showed a higher expression of inducible nitric oxide synthase in the liver, which suggests nitric oxide as a source of increased oxidative stress. A high level of etheno-DNA adducts, formed from oxyradical-induced lipid peroxidation, in liver from WD and hemochromatosis patients has been reported previously. Therefore, we exposed a wild-type p53 TK-6 lymphoblastoid cell line to 4-hydroxynonenal. an unsaturated aldehyde involved in lipid peroxidation, and observed an increase in G to T transversions at p53 codon 249 (ACC to AGT). These results are consistent with the hypothesis that the generation of oxygen/ nitrogen species and unsaturated aldehydes from iron and copper overload in hemochromatosis and WD causes mutations in the p53 tumor suppressor gene. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. MIT, Div Bioengn & Environm Hlth, Cambridge, MA 02139 USA. Univ Pittsburgh, Dept Surg, Pittsburgh, PA 15261 USA. Med Hsch Hannover, Dept Gastroenterol, D-30625 Hannover, Germany. Univ Mainz, Dept Internal Med, D-6500 Mainz, Germany. Royal Canc Hosp, Inst Canc Res, Sutton, Surrey, England. Univ Nebraska, Med Ctr, Dept Pathol, Omaha, NE 68198 USA. RP Harris, CC (reprint author), NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RI Shields, Peter/I-1644-2012 NR 56 TC 164 Z9 170 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 NOV 7 PY 2000 VL 97 IS 23 BP 12770 EP 12775 DI 10.1073/pnas.220416097 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800076 PM 11050162 ER PT J AU Salzwedel, K Berger, EA AF Salzwedel, K Berger, EA TI Cooperative subunit interactions within the oligomeric envelope glycoprotein of HIV-1: Functional complementation of specific defects in gp120 and gp41 SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; CHEMOKINE RECEPTOR; CD4 RECEPTOR; VIRAL ENTRY; SOLUBLE CD4; ANTIBODY; NEUTRALIZATION; BINDING; FUSION; ACTIVATION AB The envelope glycoprotein (Env) of HIV-1 is displayed on the surface of the virion or infected cell as an oligomer of multiple gp120/gp41 complexes. We sought to unravel the relationships between this oligomeric structure and the requirements for sequential interactions with CD4 and coreceptor (CCR5 or CXCR4). We used a quantitative cell fusion assay to examine the effects of coexpressing pairs of Envs, each nonfunctional because of a specific defect in one of the essential properties. We observed efficient fusion activity upon coexpression of two Env variants, one containing a gp41 subunit with a mutated fusion peptide and the other containing a gp120 subunit with a mutated CD4 binding site or a mismatched coreceptor specificity. We also observed fusion upon coexpression of two Env variants with distinct gp120 defects, i.e., a CD4 binding site mutation and the incorrect coreceptor specificity determinants. Coimmunoprecipitation experiments verified the efficient formation of mixed oligomers, suggesting that the observed fusion reflected subunit complementation within the oligomeric complex. These results support a model in which cooperative subunit interactions within the Env oligomer result in concerted conformational changes upon receptor binding, resulting in activation for fusion. The implications of these findings for Env function and virus neutralization are discussed. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Berger, EA (reprint author), NIAID, Viral Dis Lab, NIH, Bldg 4,Room 237, Bethesda, MD 20892 USA. NR 40 TC 51 Z9 55 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 NOV 7 PY 2000 VL 97 IS 23 BP 12794 EP 12799 DI 10.1073/pnas.230438497 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 372GK UT WOS:000165225800080 PM 11050186 ER PT J AU Blough, BE Holmquist, CR Abraham, P Kuhar, MJ Carroll, FI AF Blough, BE Holmquist, CR Abraham, P Kuhar, MJ Carroll, FI TI 3 alpha-(4-substituted phenyl)nortropane-2 beta-carboxylic acid methyl esters show selective binding at the norepinephrine transporter SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID LIGAND-BINDING; SEROTONIN TRANSPORTER; H-3 NISOXETINE; HIGH-AFFINITY; UPTAKE SITES; RADIOLIGAND; ANALOGS AB A series of 3 alpha-(4-substituted)nortropane-2 beta -carboxylic acid methyl esters was synthesized and evaluated for the ability to inhibit radioligand binding at the dopamine, serotonin, and norepinephrine transporters. 3 alpha-(4-Methylphenyl)nortropane-2 beta -carboxylic acid methyl ester (4c) was found to be selective and highly potent for the norepinephrine transporter (NET) relative to the dopamine and serotonin transporters. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 Res Triangle Inst, Res Triangle Pk, NC 27709 USA. Natl Inst Drug Abuse, Addict Res Ctr, Neurosci Branch, Baltimore, MD 21224 USA. RP Carroll, FI (reprint author), Res Triangle Inst, POB 12194,3040 Cornwallis Rd, Res Triangle Pk, NC 27709 USA. FU NIDA NIH HHS [DA05477] NR 10 TC 11 Z9 11 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 NOV 6 PY 2000 VL 10 IS 21 BP 2445 EP 2447 DI 10.1016/S0960-894X(00)00480-7 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 370AR UT WOS:000165100400012 PM 11078197 ER PT J AU Coop, A Jacobson, AE Aceto, MD Harris, LS Traynor, JR Woods, JH Rice, KC AF Coop, A Jacobson, AE Aceto, MD Harris, LS Traynor, JR Woods, JH Rice, KC TI N-cyclohexylethyl-N-noroxymorphindole: A mu-opioid preferring analogue of naltrindole SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID ANTAGONIST ACTIVITIES; RECEPTOR ANTAGONIST; SELECTIVITY; NALTREXONE; BINDING; DEPENDENCE; TOLERANCE; MORPHINE; AFFINITY; AGONISTS AB The position of the indole in the indolomorphinans, which includes the delta opioid antagonist naltrindole, is considered to be responsible for the delta opioid selectivity for this class of ligands. Herein is described the N-cyclohexylethyl substituted N-nor-derivative, which is shown to be mu preferring. Thus, the nature of the N-substituent is equally important to the receptor selectivity for this class of ligands. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 Univ Maryland, Sch Pharm, Dept Pharmaceut Sci, Baltimore, MD 21201 USA. NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol, Richmond, VA 23298 USA. Univ Michigan, Sch Med, Dept Pharmacol, Ann Arbor, MI 48109 USA. RP Coop, A (reprint author), Univ Maryland, Sch Pharm, Dept Pharmaceut Sci, 20 N Pine St, Baltimore, MD 21201 USA. NR 19 TC 7 Z9 7 U1 0 U2 1 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 NOV 6 PY 2000 VL 10 IS 21 BP 2449 EP 2451 DI 10.1016/S0960-894X(00)00479-0 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 370AR UT WOS:000165100400013 PM 11078198 ER PT J AU Tisdale, JF Dunn, DE Geller, N Plante, M Nunez, O Dunbar, CE Barrett, AJ Walsh, TJ Rosenfeld, SJ Young, NS AF Tisdale, JF Dunn, DE Geller, N Plante, M Nunez, O Dunbar, CE Barrett, AJ Walsh, TJ Rosenfeld, SJ Young, NS TI High-dose cyclophosphamide in severe aplastic anaemia: a randomised trial SO LANCET LA English DT Article ID BONE-MARROW TRANSPLANTATION; SAA-WORKING-PARTY; PULMONARY ASPERGILLOSIS; ANTITHYMOCYTE GLOBULIN; MULTIPLE-SCLEROSIS; CYCLOSPORINE-A; ANEMIA; IMMUNOSUPPRESSION; RECOVERY; THERAPY AB Background High-dose cyclophosphamide has been proposed as an alternative immunosuppressive agent for treatment of severe aplastic anaemia, with a response rate similar to that with regimens containing antithymocyte globulin (ATG) but neither relapse nor clonal haematological complications. We undertook a phase ill, prospective, randomised trial to compare response rates to immunosuppression with either high-dose cyclophosphamide plus ciclosporin or conventional immunosuppression with ATG plus ciclosporin in previously untreated patients. Methods Between June. 1997, and March. 2000, 31 patients were enrolled. 15 were assigned cyclophosphamide (1 h intravenous infusion of 50 mg/kg daily for 4 days) and 16 were assigned ATG (40 mg/kg daily for 4 days); both groups received ciclosporin, initially at 12 mg/kg daily with adjustment to maintain concentrations at 200-400 mug/L, for 6 months. The primary endpoint was haematological response (no longer meeting criteria for severe aplastic anaemia). The trial was terminated prematurely after three early deaths in the cyclophosphamide group. Analyses were by intention to treat. Findings Median follow-up was 21.9 months (range 1-33), There was excess morbidity in the cyclophosphamide group (invasive fungal infections, four cyclophosphamide vs no ATG patients; p=0.043) as well as excess early mortality (three deaths within the first 3 months cyclophosphamide vs no ATG patients; p=0.101). There was no significant difference at 6 months after treatment in the overall response rates among evaluable patients (six of 13 [46%] cyclophosphamide vs nine of 12 [75%] ATG). Interpretation A longer period of observation will be necessary to assess the secondary endpoints of relapse and late clonal complications as well as disease-free and overall survival. However, cyclophosphamide seems a dangerous choice for treatment of this disorder. given the good results achievable with standard therapy. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Tisdale, JF (reprint author), NIDDKD, Mol & Clin Hematol Branch, NIH, Bldg 10,Room 9N116, Bethesda, MD 20892 USA. NR 33 TC 82 Z9 89 U1 0 U2 3 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD NOV 4 PY 2000 VL 356 IS 9241 BP 1554 EP 1559 DI 10.1016/S0140-6736(00)03126-3 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 370QV UT WOS:000165134500011 PM 11075769 ER PT J AU Itoh, Y Law, MJ Sokoloff, L AF Itoh, Y Law, MJ Sokoloff, L TI Effects of the Na+/H+ exchanger monensin on intracellular pH in astroglia SO BRAIN RESEARCH LA English DT Article DE phosphofructokinase; cerebral glucose utilization; functional brain imaging ID CULTURED ASTROCYTES; GLUCOSE-UTILIZATION; ENERGY-METABOLISM; NEURONAL-ACTIVITY; SODIUM; DEOXYGLUCOSE AB Stimulation of astroglial glucose utilization by the Na+/H+ exchanger monensin is only partially blocked by ouabain. The present studies show that monensin also raises intracellular pH in astroglia. Because increased pn stimulates phosphofructokinase activity, the ouabain-insensitive portion of the stimulation of cerebral glucose utilization (CMRglc) appears to be due to stimulation of glycolysis by intracellular alkalinization. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIMH, Cerebral Metab Lab, Bethesda, MD 20892 USA. RP Sokoloff, L (reprint author), NIMH, Cerebral Metab Lab, Bethesda, MD 20892 USA. NR 10 TC 3 Z9 3 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD NOV 3 PY 2000 VL 882 IS 1-2 BP 226 EP 229 DI 10.1016/S0006-8993(00)02822-5 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 371DC UT WOS:000165161900028 PM 11056205 ER PT J AU Illig, KR Danilov, YP Ahmad, A Kim, CBY Spear, PD AF Illig, KR Danilov, YP Ahmad, A Kim, CBY Spear, PD TI Functional plasticity in extrastriate visual cortex following neonatal visual cortex damage and monocular enucleation SO BRAIN RESEARCH LA English DT Article DE posteromedial lateral suprasylvian; plasticity; visual cortex damage; X-pathway ID RETINAL GANGLION-CELLS; RECEPTIVE-FIELD PROPERTIES; LATERAL GENICULATE-NUCLEUS; CAT STRIATE CORTEX; X-CELLS; Y-CELLS; RETINOTOPIC ORGANIZATION; SUPRASYLVIAN CORTEX; AREAS 17; W-CELLS AB Neonatal lesions of primary visual cortex (areas 17, 18 and 19; VC) in cats lead to significant changes in the organization of visual pathways, including severe retrograde degeneration of retinal ganglion cells of the X/beta class. Cells in posteromedial lateral suprasylvian (PMLS) cortex display plasticity in that they develop normal receptive-field properties despite these changes, but they do not acquire the response properties of striate neurons that were damaged (e.g., high spatial-frequency tuning, low contrast threshold). One possibility is that the loss of X-pathway information, which is thought to underlie striate cortical properties in normal animals, precludes the acquisition of these responses by cells in remaining brain areas following neonatal VC damage. Previously, we have shown that monocular enucleation at the time of VC lesion prevents the X-/beta -cell loss in the remaining eye. The purpose of the present study was to determine whether this sparing of retinal X-cells leads to the development of striate-like response properties in PMLS cortex. We recorded the responses of PMLS neurons to visual stimuli to assess spatial-frequency tuning, spatial resolution, and contrast threshold. Results indicated that some PMLS cells in animals with a neonatal VC lesion and monocular enucleation displayed a preference for higher spatial frequencies, had higher spatial resolution, and had lower contrast thresholds than PMLS cells in cats with VC lesion alone. Taken together, these results suggest that preserving X-pathway input during this critical period leads to the addition of some X-like properties to PMLS visual responses. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Wisconsin, Sch Med, Dept Anat, Madison, WI 53706 USA. Univ Wisconsin, Sch Med, Ctr Neurosci, Madison, WI 53706 USA. NIAAA, Lab Membrane Biochem & Biophys, Rockville, MD 20852 USA. Univ Wisconsin, Dept Ophthalmol & Visual Sci, Madison, WI 53792 USA. Univ Colorado, Dept Psychol, Boulder, CO 80309 USA. RP Illig, KR (reprint author), Univ Wisconsin, Sch Med, Dept Anat, 1300 Univ Ave, Madison, WI 53706 USA. NR 46 TC 4 Z9 6 U1 1 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD NOV 3 PY 2000 VL 882 IS 1-2 BP 241 EP 250 DI 10.1016/S0006-8993(00)02902-4 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 371DC UT WOS:000165161900031 PM 11056208 ER PT J AU Zhang, YL Yao, ZJ Sarmiento, M Wu, L Burke, TR Zhang, ZY AF Zhang, YL Yao, ZJ Sarmiento, M Wu, L Burke, TR Zhang, ZY TI Thermodynamic study of ligand binding to protein-tyrosine phosphatase 1B and its substrate-trapping mutants SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ENTHALPY-ENTROPY COMPENSATION; SOLID-PHASE SYNTHESIS; TITRATION CALORIMETRY; O-PHOSPHOTYROSINE; TRANSITION-STATE; INSULIN-RECEPTOR; CRYSTAL-STRUCTURE; INHIBITOR DESIGN; FOCAL ADHESIONS; SIGNATURE MOTIF AB The binding of several phosphonodifluoromethyl phenylalanine (F(2)Pmp)-containing peptides to protein-tyrosine phosphatase 1B (PTP1B) and its substrate-trapping mutants (C215S and D181A) has been studied using isothermal titration calorimetry. The binding of a high affinity ligand, Ac-Asp-Ala-Asp-Glu-F(2)Pmp-Leu-NH2, to PTP1B (K-d = 0.24 muM) is favored by both enthalpic and entropic contributions. Disruption of ionic interactions between the side chain of Arg-47 and the N-terminal acidic residues reduces the binding affinity primarily through the reduction of the T DeltaS term. The role of Arg-47 may be to maximize surface contact between PTP1B and the peptide, which contributes to high affinity binding. The active site Cys-215-->Ser mutant PTP1B binds ligands with the same affinity as the wild-type enzyme. However, unlike wild-type PTP1B, peptide binding to C215S is predominately driven by enthalpy change, which likely results from the elimination of the electrostatic repulsion between the thiolate anion and the phosphonate group. The increased enthalpic contribution is offset by reduction in the binding entropy, which may be the result of increased entropy of the unbound protein caused by this mutation. The general acid-deficient mutant D181A binds the peptide 5-fold tighter than the C215S mutant, consistent with the observation that the Asp to Ala mutant is a better "substrate-trapping" reagent than C215S, The increased binding affinity for D181A as compared with the wildtype PTP1B results primarily from an increase in the DeltaH of binding in the mutant, which may be related to decreased electrostatic repulsion between the phosphate moiety and PTP1B, These results have important implications for the design of high affinity PTP1B inhibitors. C1 Yeshiva Univ Albert Einstein Coll Med, Dept Mol Pharmacol, Bronx, NY 10461 USA. Yeshiva Univ Albert Einstein Coll Med, Dept Biochem, Bronx, NY 10461 USA. NCI, Med Chem Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Zhang, ZY (reprint author), Yeshiva Univ Albert Einstein Coll Med, Dept Mol Pharmacol, 1300 Morris Pk Ave, Bronx, NY 10461 USA. RI Burke, Terrence/N-2601-2014 FU NIGMS NIH HHS [5T32-GM-07491, GM-55242] NR 63 TC 40 Z9 42 U1 1 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34205 EP 34212 DI 10.1074/jbc.M004490200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300030 PM 10952978 ER PT J AU Ray, K Hauschild, BC AF Ray, K Hauschild, BC TI Cys-140 is critical for metabotropic glutamate receptor-1 dimerization SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PUTATIVE PHEROMONE RECEPTORS; HUMAN CALCIUM RECEPTOR; TERMINAL EXTRACELLULAR DOMAIN; CELL-SURFACE EXPRESSION; LIGAND-BINDING; SIGNAL-TRANSDUCTION; MULTIGENE FAMILY; CLONING; IDENTIFICATION; PURIFICATION AB Metabotropic glutamate receptor 1 (mGluR1) expresses at the cell surface as disulfide-linked dimers and can be reduced to monomers with sulfhydryl reagents. To identify the dimerization domain, we transiently expressed in HEK-293 cells a truncated version of mGluR1 (RhodC-R1) devoid of the extracellular domain (ECD), RhodC-R1 was a monomer in the absence or presence of the reducing agents, suggesting that dimerization occurs via the ECD, To identify cysteine residues involved in dimerization within the ECD, cysteine to serine point mutations were made at three cysteines within the amino-terminal half of the ECD, A mutation at positions Cys-67, Cys-109, and Cys-140 all resulted in significant amounts of monomers in the absence of reducing agents. The monomeric C67S and C109S mutants were not properly glycosylated, failed to reach the cell surface, and showed no glutamate response, indicating that these mutant receptors were improperly folded and/or processed and thus retained intracellularly, In contrast, the monomeric C140S mutant was properly glycosylated, processed, and expressed at the cell surface. Phosphoinositide hydrolysis assay showed that the glutamate response of the C140S mutant receptor was similar to the wild type receptor. Substitution of a cysteine for Ser-129, Lys-134, Asp-143, and Thr-146 on the C140S mutant background restored receptor dimerization. Taken together, the results suggest that Cys-140 contributes to intermolecular disulfide linked dimerization of mGluR1. C1 NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Ray, K (reprint author), Natl Inst Deafness & Other Commun Disorders, Lab Signal Transduct, 5 Res Ct, Rockville, MD 20850 USA. NR 31 TC 59 Z9 60 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34245 EP 34251 DI 10.1074/jbc.M005581200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300035 PM 10945991 ER PT J AU Chen, LY Lin, BC Pan, CJ Hiraiwa, H Chou, JY AF Chen, LY Lin, BC Pan, CJ Hiraiwa, H Chou, JY TI Structural requirements for the stability and microsomal transport activity of the human glucose 6-phosphate transporter SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GLYCOGEN-STORAGE-DISEASE; PUTATIVE GLUCOSE-6-PHOSPHATE TRANSLOCASE; TRANSMEMBRANE TOPOLOGY; CHROMOSOME 11Q23; GENE; IB; 1B; MUTATION; PROTEIN AB Deficiencies in glucose 6-phosphate (G6P) transporter (G6PT), a 10-helical endoplasmic reticulum transmembrane protein of 429 amino acids, cause glycogen storage disease type Ib. To date, only three missense mutations in G6PT have been shown to abolish microsomal G6P transport activity. Here, we report the results of structure-function studies on human G6PT and demonstrate that 15 missense mutations and a codon deletion (Delta F93) mutation abolish microsomal G6P uptake activity and that two splicing mutations cause exon skipping. While most missense mutants support the synthesis of G6PT protein similar to that of the wild-type transporter, immunoblot analysis shows that G20D, Delta F93, and I278N mutations, located in helix 1, 2, and 6, respectively, destabilize the G6PT. Further, we demonstrate that G6PT mutants lacking an intact helix 10 are misfolded and undergo degradation within cells. Moreover, amino acids 415-417 in the cytoplasmic tail of the carboxyl-domain, extending from helix 10, also play a critical role in the correct folding of the transporter. However, the last 12 amino acids of the cytoplasmic tail play no essential role(s) in functional integrity of the G6PT. Our results, for the first time, elucidate the structural requirements for the stability and transport activity of the G6PT protein. C1 NICHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Chou, JY (reprint author), NICHD, Heritable Disorders Branch, NIH, Bldg 10,Rm 9S241, Bethesda, MD 20892 USA. RI Lin, Baochuan/A-8390-2009 OI Lin, Baochuan/0000-0002-9484-0785 NR 30 TC 20 Z9 20 U1 1 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34280 EP 34286 DI 10.1074/jbc.M006439200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300040 PM 10940311 ER PT J AU Pedersen, LC Tsuchida, K Kitagawa, H Sugahara, K Darden, TA Negishi, M AF Pedersen, LC Tsuchida, K Kitagawa, H Sugahara, K Darden, TA Negishi, M TI Heparan/chondroitin sulfate biosynthesis - Structure and mechanism of human glucuronyltransferase I SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROTEIN LINKAGE-REGION; HEPARAN-SULFATE; N-ACETYLGALACTOSAMINE; MAST-CELLS; GLYCOSYLTRANSFERASES; DROSOPHILA; ENZYME; DOMAIN; PROTEOGLYCANS; ENCODES AB Human beta1,3-glucuronyltransferase I (GlcAT-I) is a central enzyme in the initial steps of proteoglycan synthesis. GlcAT-I transfers a glucuronic acid moiety from the uridine diphosphate-glucuronic acid (UDP-GlcUA) to the common linkage region trisaccharide Gal beta1-3Gal beta1-4Xyl covalently bound to a Ser residue at the glycosaminylglycan attachment site of proteoglycans, We have now determined the crystal structure of GlcAT-I at 2.3 Angstrom in the presence of the donor substrate product UDP, the catalytic Mn2+ ion, and the acceptor substrate analog Gal beta1-3Gal beta1-4Xyl, The enzyme is a alpha/beta protein with two subdomains that constitute the donor and acceptor substrate binding site. The active site residues lie in a cleft extending across both subdomains in which the trisaccharide molecule is oriented perpendicular to the UDP. Residues Glu(227), Asp(252), and Glu(281) dictate the binding orientation of the terminal Gal-2 moiety. Residue Glu(281) is in position to function as a catalytic base by deprotonating the incoming 3-hydroxyl group of the acceptor. The conserved DXD motif (Asp(191), Asp(195), Asp(196)) has direct interaction with the ribose of the UDP molecule as well as with the Mn2+ ion. The key residues involved in substrate binding and catalysis are conserved in the glucuronyltransferase family as well as of her glycosyltransferases. C1 NIEHS, Reprod & Dev Toxicol Lab, Pharmacogenet Sect, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. Kobe Pharmaceut Univ, Dept Biochem, Higashinada Ku, Kobe, Hyogo 6858858, Japan. RP Negishi, M (reprint author), NIEHS, Reprod & Dev Toxicol Lab, Pharmacogenet Sect, NIH, Res Triangle Pk, NC 27709 USA. NR 32 TC 150 Z9 152 U1 0 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34580 EP 34585 DI 10.1074/jbc.M007399200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300079 PM 10946001 ER PT J AU Buxton, DB Adelstein, RS AF Buxton, DB Adelstein, RS TI Calcium-dependent threonine phosphorylation of nonmuscle myosin in stimulated RBL-2H3 mast cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROTEIN-KINASE-C; II HEAVY-CHAIN; BASOPHILIC LEUKEMIA-CELLS; SPHINGOSINE KINASE; HISTAMINE-RELEASE; ANTIGEN; EXOCYTOSIS; SECRETION; RECEPTOR; MTS1 AB Stimulation of RBL-2H3 mi mast cells through the IgE receptor with antigen, or through a G protein-coupled receptor with carbachol, leads to the rapid appearance of phosphothreonine in nonmuscle myosin heavy chain ZI-A (NMHC-IIA). We demonstrate that this results from phosphorylation of Thr-1940 by calcium/calmodulin-dependent protein kinase II (CaM kinase II), activated by increased intracellular calcium. The phosphorylation site in rodent NMHC-IIA was localized to the carboxyl terminus of NMHC-IIA distal to the coiled-coil region, and,identified as Thr-1940 by site-directed mutagenesis. A fusion protein containing the NMHC-IIA carboxyl terminus was phosphorylated by GaM kinase II in vitro, while mutation of Thr-1940 to Ala eliminated phosphorylation. In contrast to rodents, in humans Thr-1940 is replaced by Ala, and human NMHC-IIA fusion protein was not phosphorylated by CaM kinase II unless Ala-->1940 was mutated to Thr, Similarly, co-transfected Ala-->Thr-1940 human NMHC-IIA was phosphorylated by activated CaM: kinase II in HeLa cells, while wild type was not. In RBL-2H3 mi cells, inhibition of CaM kinase II decreased Thr-1940 phosphorylation, and inhibited release of the secretory granule marker hexosaminidase in response to carbachol but not to antigen. These data indicate a role for CaM kinase stimulation and resultant threonine phosphorylation of NMHC-IIA in RBL-2H3 mi cell activation. C1 NHLBI, Mol Cardiol Lab, NIH, Bethesda, MD 20892 USA. RP Buxton, DB (reprint author), NHLBI, Mol Cardiol Lab, NIH, Bldg 10,Rm 8N202,10 Ctr Dr, Bethesda, MD 20892 USA. OI Buxton, Denis/0000-0003-3077-6435; Adelstein, Robert/0000-0002-8683-2144 NR 40 TC 16 Z9 16 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34772 EP 34779 DI 10.1074/jbc.M004996200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300103 PM 10945986 ER PT J AU Moll, JR Olive, M Vinson, C AF Moll, JR Olive, M Vinson, C TI Attractive interhelical electrostatic interactions in the proline- and acidic-rich region (PAR) leucine zipper subfamily preclude heterodimerization with other basic leucine zipper subfamilies SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID COILED-COIL; TRANSCRIPTION FACTOR; DNA-BINDING; CRYSTAL-STRUCTURE; C-FOS; EXPRESSION; SPECIFICITY; LEUKEMIA; GENE; PROTEIN AB Basic region-leucine zipper (B-ZIP) proteins homo- or heterodimerize to bind sequence-specific double-stranded DNA. We present circular dichroism (CD) thermal denaturation data on vitellogenin promoter-binding protein (VBP), a member of the PAR subfamily of E-ZIP proteins that also includes thyroid embryonic factor, hepatocyte leukemia factor, and albumin site D-binding protein. VBP does not heterodimerize with B-ZIP domains from C/EBP alpha, JUND, or FOS, We describe a dominant negative protein, A-VEP, that contains the VBP leucine zipper and an acidic amphipathic protein sequence that replaces the basic region critical for DNA binding. The acidic extension forms a coiled coil structure with the VEP basic region in the VBP A-VBP heterodimer, This new alpha -helical structure extends the leucine zipper N-terminally, stabilizing the complex by 2.0 kcal/mol, A-VBP abolishes DNA binding of VBP in an equimolar competition assay, but does not affect DNA binding even at 100-fold excess of CREB, C/EBP alpha, or FOS/JUND, Likewise, proteins containing the acidic extension appended to seven other leucine zippers do not inhibit VBP DNA binding. We show that conserved g <-> e' or i, i' + 5 salt bridges are sufficient to confer specificity to VBP by mutating the C/EBP alpha leucine zipper to contain the g <-> e' salt bridges that characterize VBP, A-VBP heterodimerizes with this mutant C/EBP, preventing it from binding to DNA These conserved g <-> e' electrostatic interactions define the specificity of the PAR subfamily of B-ZIP proteins and preclude interaction with other B-ZIP subfamilies. C1 NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. INSERM, U441, F-33600 Pessac, France. RP Vinson, C (reprint author), NCI, Lab Metab, NIH, Bldg 37,Rm 4D06, Bethesda, MD 20892 USA. NR 29 TC 30 Z9 30 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD NOV 3 PY 2000 VL 275 IS 44 BP 34826 EP 34832 DI 10.1074/jbc.M004545200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 369YL UT WOS:000165095300110 PM 10942764 ER PT J AU Koonin, EV Aravind, L AF Koonin, EV Aravind, L TI Dynein light chains of the roadblock/LC7 group belong to an ancient protein superfamily implicated in NTPase regulation SO CURRENT BIOLOGY LA English DT Letter ID MYXOCOCCUS-XANTHUS; TRANSPORT; MOTILITY C1 NIH, Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. RP Koonin, EV (reprint author), NIH, Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. NR 12 TC 45 Z9 47 U1 1 U2 5 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD NOV 2 PY 2000 VL 10 IS 21 BP R774 EP R776 DI 10.1016/S0960-9822(00)00774-0 PG 3 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 372JG UT WOS:000165230100003 PM 11084347 ER PT J AU Hoffman, BT Kopajtic, T Katz, JL Newman, AH AF Hoffman, BT Kopajtic, T Katz, JL Newman, AH TI 2D QSAR modeling and preliminary database searching for dopamine transporter inhibitors using genetic algorithm variable selection of Molconn Z descriptors SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID MOLECULAR-FIELD ANALYSIS; PARTIAL LEAST-SQUARES; RECEPTOR LIGANDS; COCAINE; ANALOGS; AFFINITY; INFORMATION; DERIVATIVES; BINDING; COMBINATION AB In light, of the chronic problem of abuse of the controlled substance cocaine, we have investigated novel approaches toward both understanding the activity of inhibitors of the dopamine transporter (DAT) and identifying novel inhibitors that may be of therapeutic potential. Our most recent studies toward these ends have made use of two-dimensional (2D) quantitative structure-activity relationship (QSAR) methods in order to develop predictive models that correlate structural features of DAT ligands to their biological activities. Specifically, we have adapted the method of genetic algorithms-partial least squares (GA-PLS) (Cho et al. J. Comput.-Aided Mel. DES., submitted) to the task of variable selection of the descriptors generated by the software Molconn Z. As the successor to the program Molconn X, which generated 462 descriptors, Molconn Z provides 749 chemical descriptors. By employing genetic algorithms in optimizing the inclusion of predictive descriptors, we have successfully developed a robust model of the DAT affinities of 70 structurally diverse DAT ligands. This model, with an exceptional q(2) value of 0.85, is nearly 25% more accurate in predictive value than a comparable model derived from Molconn X-derived descriptors (q(2) = 0.69). Utilizing activity-shuffling validation methods, we have demonstrated the robustness of both this DAT inhibitor model and our QSAR method. Moreover, we have extended this method to the analysis of dopamine D-1 antagonist affinity and serotonin ligand activity, illustrating the significant improvement in q(2) for a variety of data sets. Finally, we have employed our method in performing a search of the National Cancer Institute database based upon activity predictions from our DAT model. We report the preliminary results of this search, which has yielded five compounds suitable for lead development as novel DAT inhibitors. C1 Natl Inst Drug Abuse, Intramural Res Program, Med Chem Sect, Baltimore, MD 21224 USA. Natl Inst Drug Abuse, Intramural Res Program, Psychobiol Sect, Baltimore, MD 21224 USA. RP Newman, AH (reprint author), Natl Inst Drug Abuse, Intramural Res Program, Med Chem Sect, Baltimore, MD 21224 USA. OI Katz, Jonathan/0000-0002-1068-1159 NR 41 TC 47 Z9 47 U1 0 U2 2 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 NOV 2 PY 2000 VL 43 IS 22 BP 4151 EP 4159 DI 10.1021/jm990472s PG 9 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 371UX UT WOS:000165198600012 PM 11063611 ER PT J AU Ma, JY Katz, E Kyle, DE Ziffer, H AF Ma, JY Katz, E Kyle, DE Ziffer, H TI Syntheses and antimalarial activities of 10-substituted deoxoartemisinins SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID ARTEMISININ; ANALOGS; DERIVATIVES; CONCISE AB Two series of 10-substituted deoxoartemisinin derivatives have been synthesized. The first employed the reaction of dihydroartemisinin acetate with several silyl enol ethers in the presence of titanium tetrachloride. The second utilized the reaction of 10-(2-oxoethyl)deoxoartemisinin with several Grignard reagents. The in vitro antimalarial activities of both series were determined against two drug-resistant clones of P. falciparum. The activities of 13 beta and 15 beta were 5-7 times greater than that of artemisinin. C1 NIDDK, Phys Chem Lab, NIH, Bethesda, MD 20892 USA. NIDDK, Bioorgan Chem Lab, Bethesda, MD 20892 USA. Walter Reed Army Med Ctr, Walter Reed Army Inst Res, Dept Parasitol, Washington, DC 20307 USA. RP Ziffer, H (reprint author), NIDDK, Phys Chem Lab, NIH, Bldg 5,Rm B1-31, Bethesda, MD 20892 USA. NR 13 TC 35 Z9 37 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 NOV 2 PY 2000 VL 43 IS 22 BP 4228 EP 4232 DI 10.1021/jm0001951 PG 5 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 371UX UT WOS:000165198600020 PM 11063619 ER PT J AU Chow, WH Gridley, G Fraumeni, JF Jarvholm, B AF Chow, WH Gridley, G Fraumeni, JF Jarvholm, B TI Obesity, hypertension, and the risk of kidney cancer in men. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID RENAL-CELL CANCER; GROWTH-FACTOR; PREDICTORS; CARCINOMA; HORMONE AB Background: Obesity and hypertension have been implicated as risk factors for the development of renal-cell cancer. Methods: We examined the health records of 363,992 Swedish men who underwent at least one physical examination from 1971 to 1992 and were followed until death or the end of 1995. Men with cancer (renal-cell cancer in 759 and renal-pelvis cancer in 136) were identified by cross-linkage of data with the nationwide Swedish Cancer Registry. Poisson regression analysis was used to estimate relative risks, with adjustments for age, smoking status, body-mass index, and diastolic blood pressure. Results: As compared with men in the lowest three eighths of the cohort for body-mass index, men in the middle three eighths had a 30 to 60 percent greater risk of renal-cell cancer, and men in the highest two eighths had nearly double the risk (P for trend, <0.001). There was also a direct association between higher blood pressures and a higher risk of renal-cell cancer (P for trend, <0.001 for diastolic pressure; P for trend, 0.007 for systolic pressure). After the first five years of follow-up had been excluded to reduce possible effects of preclinical disease, the risk of renal-cell cancer was still consistently higher in men with a higher body-mass index or higher blood pressure. At the sixth-year follow-up, the risk rose further with increasing blood pressures and decreased with decreasing blood pressures, after adjustment for base-line measurements. Men who were current or former smokers had a greater risk of both renal-cell cancer and renal-pelvis cancer than men who were not smokers. There was no relation between body-mass index or blood pressure and the risk of renal-pelvis cancer. Conclusions: Higher body-mass index and elevated blood pressure independently increase the long-term risk of renal-cell cancer in men. A reduction in blood pressure lowers the risk. (N Engl J Med 2000;343:1305-11.) (C) 2000, Massachusetts Medical Society. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Umea Univ, Dept Publ Hlth & Clin Med, Umea, Sweden. RP Chow, WH (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS 8100, Bethesda, MD 20892 USA. FU NCI NIH HHS [N02-CP-71100] NR 27 TC 313 Z9 325 U1 2 U2 14 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 NOV 2 PY 2000 VL 343 IS 18 BP 1305 EP 1311 DI 10.1056/NEJM200011023431804 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 368RP UT WOS:000090131600004 PM 11058675 ER PT J AU Tatum, JL Hoffman, JM AF Tatum, JL Hoffman, JM TI Imaging drug development SO ACADEMIC RADIOLOGY LA English DT Article C1 NCI, Biomed Imaging Program, DCTD, BIP, Bethesda, MD 20892 USA. RP Tatum, JL (reprint author), NCI, Biomed Imaging Program, DCTD, BIP, EPN-600,6130 Execut Blvd,MSC 7412, Bethesda, MD 20892 USA. NR 0 TC 7 Z9 7 U1 0 U2 1 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 NOV PY 2000 VL 7 IS 11 BP 1007 EP 1008 PG 2 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 369YB UT WOS:000165094400015 PM 11089703 ER PT J AU Dennis, ML Perl, HI Huebner, RB McLellan, AT AF Dennis, ML Perl, HI Huebner, RB McLellan, AT TI Twenty-five strategies for improving the design, implementation and analysis of health services research related to alcohol and other drug abuse treatment SO ADDICTION LA English DT Article; Proceedings Paper CT Conference of the National-Institute-on-Alcohol-Abuse-and-Alcoholism (NIAAA) CY APR 19-20, 1999 CL ROCKVILLE, MARYLAND SP Natl Inst Alcohol Abuse & Alcoholism ID ADOLESCENT SUBSTANCE-ABUSE; RANDOMIZED EXPERIMENTS; CLINICAL-TRIALS; VALIDITY; ASSIGNMENT; ADDICTION; SEVERITY; OUTCOMES; QUALITY; ISSUES AB While some aspects of addiction can be studied in laboratory or controlled settings, the study of long-term recovery management and the health services that support it requires going our into the community and dealing with populations and systems that are much more diverse and less under our control. This in turn raises many methodological challenges for the health service researchers studying alcohol and other drug abuse treatment. This paper identifies some of these challenges related to the design, measurement, implementation and effectiveness of health services research. It then recommends 25 strategies (and key primers) for addressing them: (1) identifying in advance all stakeholders and issues; (2) developing conceptual models of intervention and context; (3) identifying the population to whom the conclusions will be generalized; (4) matching the research design to the question; (5) conducting randomized experiments only when appropriate and necessary; (6) balancing methodological and treatment concerns; (7) prioritizing analysis plans and increasing design sensitivity, (8) combining qualitative and quantitative methods; (9) identifying the four basic types of measures needed; (10) identifying and using standardized measures; (11) carefully balancing measurement selection and modification; (12) developing and evaluating modified and new measures when necessary; (13) identifying and tracking major clinical subgroups; (14) measuring and analyzing the actual pattern of services received; (15) incorporating implementation checks into the design; (16) incorporating baseline measures into the intervention; (17) monitoring implementation and dosage as a form of quality assurance; (18) developing procedures early to facilitate tracking and follow-up of study participants; (19) using more appropriate representations of the actual experiment; (20) using appropriate and sensitive standard deviation terms; (21) partialing out variance due to design or known sources prior to estimating experimental effect sizes; (22) using dimensional, interval and ratio measures to increase sensitivity to change; (23) using path or structural equation models; (24) integrating qualitative and quantitative analysis into reponing; and (25) using quasi-experiments, economic or organizational studies to answer other likely policy questions. Most of these strategies have been tried and tested in this and other areas, bur are not widely used. Improving the state of the art of health services research and bridging the gap between research and practice do not depend upon using the most advanced methods, but rather upon using the most appropriate methods. C1 Chestnut Hlth Syst, Bloomington, IL 61701 USA. NIAAA, Bethesda, MD USA. Univ Penn, Philadelphia, PA 19104 USA. RP Dennis, ML (reprint author), Chestnut Hlth Syst, 720 W Chestnut, Bloomington, IL 61701 USA. FU NIAAA NIH HHS [AA10368]; NIDA NIH HHS [P50-DA06990] NR 118 TC 33 Z9 33 U1 2 U2 10 PU CARFAX PUBLISHING PI BASINGSTOKE PA RANKINE RD, BASINGSTOKE RG24 8PR, HANTS, ENGLAND SN 0965-2140 J9 ADDICTION JI Addiction PD NOV PY 2000 VL 95 SU 3 BP S281 EP S308 PG 28 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 380ZL UT WOS:000165737800002 PM 11132359 ER PT J AU Perl, HI Dennis, ML Huebner, RB AF Perl, HI Dennis, ML Huebner, RB TI State-of-the-art methodologies in alcohol-related health services research SO ADDICTION LA English DT Editorial Material AB Many of the failures to replicate clinical findings of treatment efficacy in more realistic field and community settings can be attributed to inappropriate research designs and other methodological shortcomings. In order to increase research designers' awareness of existing methodologies that may be better suited to answer the critical questions inherent in health services research on alcohol-related issues, the National Institute on Alcohol Abuse and Alcoholism (NIAAA) convened an expert conference with three specific goals: (1) to identify the critical issues involved in alcohol services research; (2) to develop a primer that explicated each key area; and (3) to compile the resulting primers into an accessible resource for researchers, policy makers and consumers. The 9 papers in this special supplement are the product of that conference and are organized broadly around three phases of the research process: study design and implementation, data collection and use, and the analysis and interpretation of data. A final summary paper discusses the issues and offers a synthesis of key themes as well as some direction for the future. C1 NIAAA, Hlth Serv Res Branch, Div Clin & Prevent Res, Bethesda, MD 20892 USA. Chestnut Hlth Syst, Bloomington, IL USA. RP Perl, HI (reprint author), NIAAA, Hlth Serv Res Branch, Div Clin & Prevent Res, 6000 Execut Blvd,MSC 7003,Suite 505, Bethesda, MD 20892 USA. NR 7 TC 7 Z9 7 U1 0 U2 0 PU CARFAX PUBLISHING PI BASINGSTOKE PA RANKINE RD, BASINGSTOKE RG24 8PR, HANTS, ENGLAND SN 0965-2140 J9 ADDICTION JI Addiction PD NOV PY 2000 VL 95 SU 3 BP S275 EP S280 PG 6 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 380ZL UT WOS:000165737800001 PM 11132358 ER PT J AU Leischow, SJ Ranger-Moore, J Lawrence, D AF Leischow, SJ Ranger-Moore, J Lawrence, D TI Addressing social and cultural disparities in tobacco use SO ADDICTIVE BEHAVIORS LA English DT Article; Proceedings Paper CT International Symposium on Addictions 2000 CY SEP 22-24, 2000 CL HYANNIS, MASSACHUSETTS DE tobacco; social disparity; smoking; risk factors ID DRUG-ABUSE PREVENTION; CIGARETTE-SMOKING; POPULATION; BEHAVIORS; ADULTS; QUIT AB Disparities in smoking as a function of ethnicity exist, but part of that disparity appears to be mediated by education level. Smoking prevalence differences by ethnicity disappear or decrease substantially when controlling for education level. Nonetheless, ethnicity remains an important consideration when striving to understand the mechanisms associated with smoking, as well as in the development of smoking prevention and treatment programs. In this paper, the complex interrelationships among ethnicity, social class, and smoking are discussed in brief, and a framework for developing a systematic means of addressing social disparities in tobacco use is described. (C) 2000 Elsevier Science Ltd. C1 NCI, Div Canc Control & Populat Sci, Tobacco Control Res Branch, Bethesda, MD 20892 USA. Univ Arizona, Tucson, AZ 85721 USA. RP Leischow, SJ (reprint author), NCI, Div Canc Control & Populat Sci, Tobacco Control Res Branch, 6130 Execut Blvd,EPN 4038, Bethesda, MD 20892 USA. NR 33 TC 10 Z9 10 U1 1 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0306-4603 J9 ADDICT BEHAV JI Addict. Behav. PD NOV-DEC PY 2000 VL 25 IS 6 BP 821 EP 831 DI 10.1016/S0306-4603(00)00123-4 PG 11 WC Psychology, Clinical; Substance Abuse SC Psychology; Substance Abuse GA 374KH UT WOS:000165344000003 PM 11125773 ER PT J AU Jacobson, S Franchini, G Jeang, KT AF Jacobson, S Franchini, G Jeang, KT TI HTLV Molecular Biology and Pathogenesis - March 17-19, 2000 - Introduction SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Editorial Material C1 NINDS, Bethesda, MD 20892 USA. NCI, Bethesda, MD 20892 USA. NIAID, Bethesda, MD 20892 USA. RP Jacobson, S (reprint author), NINDS, Bldg 36,Rm 4D04, Bethesda, MD 20892 USA. RI Jeang, Kuan-Teh/A-2424-2008 NR 0 TC 0 Z9 0 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 NOV 1 PY 2000 VL 16 IS 16 BP 1581 EP 1581 DI 10.1089/08892220050192985 PG 1 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900001 ER PT J AU Nicot, C Opavsky, R Mahieux, R Johnson, JM Brady, JN Wolff, L Franchini, G AF Nicot, C Opavsky, R Mahieux, R Johnson, JM Brady, JN Wolff, L Franchini, G TI Tax oncoprotein trans-represses endogenous B-myb promoter activity in human T cells SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID VIRUS TYPE-1 TAX; LEUKEMIA; GENE; EXPRESSION; PROTEIN; PROLIFERATION; COACTIVATOR; APOPTOSIS; CBP AB The B-myb gene was identified on the basis of its homology with the protooncogene c-myb, homolog of the avian myeloblastosis virus (AMV) and avian leukemia virus (E26) transforming genes. Several studies using antisense constructs or antisense oligonucleotides as well as overexpression experiments suggest that B-Myb plays an important role in the transition from G(1) to S phase of the cell cycle and that B-Myb expression is cell cycle regulated. We have previously demonstrated that the human T cell lymphotropic virus type 1 (HTLV-1) trans-activator Tax is able to repress transcription from c-myb promoter reporter constructs as well as from the endogenous c-myb promoter in human T cells and that this effect is mediated through inhibition of the cMyb trans-activating functions. Here we report that both HTLV-1 as well as HTLV-2 Tax proteins inhibit cMyb trans-activation in mouse embryo fibroblasts (MEFs). In addition to c-Myb, B-Myb expression is also markedly downregulated in HTLV-1-transformed cells at both RNA and protein levels. Furthermore, by using a Jurkat T cell line stably transfected with a tax gene driven by a cadmium-inducible promoter (JPX9), we were able to demonstrate that Tax directly represses the endogenous B-myb promoter in T cells. Because c-Myb and R-Myb have been involved in cell cycle progression, our results suggest that Tax, by repressing both c-Myb and B-Myb endogenous promoters, may bypass their requirement for cell cycle progression in HTLV-1-transformed T cells. C1 NCI, Basic Res Labs, Div Basic Sci, NIH, Bethesda, MD 20892 USA. NIH, Cellular Oncol Lab, Bethesda, MD 20892 USA. NIH, Lab Receptor Biol & Gene Express, Bethesda, MD 20892 USA. RP Nicot, C (reprint author), NCI, Basic Res Labs, Div Basic Sci, NIH, 9000 Rockville Pike,Bldg 41,Room C303, Bethesda, MD 20892 USA. NR 13 TC 15 Z9 15 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 NOV 1 PY 2000 VL 16 IS 16 BP 1629 EP 1632 DI 10.1089/08892220050193065 PG 4 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900009 PM 11080802 ER PT J AU Iha, H Kasai, T Kibler, KV Iwanaga, Y Tsurugi, K Jeang, KT AF Iha, H Kasai, T Kibler, KV Iwanaga, Y Tsurugi, K Jeang, KT TI Pleiotropic effects of HTLV type 1 tax protein on cellular metabolism: Mitotic checkpoint abrogation and NF-kappa B activation SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID VIRUS TYPE-I; ZINC-FINGER PROTEIN; NECROSIS-FACTOR-ALPHA; LONG TERMINAL REPEAT; HUMAN LUNG CANCERS; LEUKEMIA LYMPHOMA; TRANSCRIPTION FACTORS; GENE-EXPRESSION; INFECTED-CELLS; A20 INTERACTS AB Tax protein expressed by human T cell leukemia virus type 1 (HTLV-1) is a strong trans-activator of its own LTR promoter; it also affects the function of multiple cellular genes involved in cell cycle control and transcription. One way in which Tax exerts its pleiotropic effects is through protein-protein interaction with cellular cofactors. By using yeast two-hybrid technology, we have isolated several cellular proteins that bind to Tax. Two of these are MAD1, a mitotic checkpoint control protein, and TXBP151, a suppressor of tumor necrosis factor alpha -induced apoptosis, Here we discuss findings describing the role of MAD1 in exit of cells from mitosis and TXBP151 in NF-kappaB activation. C1 NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. Ymanashi Med Univ, Biochem Lab, Yamanashi 4093898, Japan. RP Iha, H (reprint author), NIAID, Mol Microbiol Lab, NIH, 9000 Rockville Pike,Bldg 4,Room 304, Bethesda, MD 20892 USA. RI Jeang, Kuan-Teh/A-2424-2008; OI Iha, Hidekatsu/0000-0002-0999-5636 NR 44 TC 20 Z9 20 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 NOV 1 PY 2000 VL 16 IS 16 BP 1633 EP 1638 DI 10.1089/08892220050193074 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900010 PM 11080803 ER PT J AU Pise-Masison, CA Mahieux, R Radonovich, M Jiang, H Duvall, J Guillerm, C Brady, JN AF Pise-Masison, CA Mahieux, R Radonovich, M Jiang, H Duvall, J Guillerm, C Brady, JN TI Insights into the molecular mechanism of p53 inhibition by HTLV type 1 Tax SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID CELL LEUKEMIA-VIRUS; NF-KAPPA-B; CREB-BINDING-PROTEIN; HUMAN T-LYMPHOCYTES; WILD-TYPE P53; TRANSFORMING PROTEIN; GENE-EXPRESSION; IKK-ALPHA; ACTIVATION; KINASE AB The p53 protein plays a pivotal role in transmitting signals from many forms of genotoxic stress to genes and factors that control aspects of the cell cycle and death. Although mutated in approximately 60% of all human cancers, only a minority of human T-lymphotropic virus type 1 (HTLV-1)-transformed cells carry p53 mutations. Nevertheless, the p53 protein in HTLV-1-transformed cells is functionally inactive. We have previously demonstrated that the HTLV-1 Tax protein can inhibit p53 trans-activation function. Tax does not accomplish this by directly binding to p53, but rather by a unique mechanism that includes constitutive phosphorylation of p53 at Ser-15 and Ser-392. Analysis of Tax mutants in lymphocytes demonstrates that Tax-induced p53 inhibition correlates with the ability of Tax to activate NP-kappaB, but not p300 binding or CREB trans-activation. Consistent with these results, expression of the I-kappaB alpha (S32,36A) mutant that blocks NF-kappaB activation blocks Tax-mediated p53 inhibition. We further demonstrate the importance of Tax activation of NF-kappaB in p53 inhibition, using p65 knockout (KO) mouse embryo fibroblasts (MEFs). In the absence of p65 Tax could not inhibit p53. Tax does activate IKK beta in the p65 KO MEFs, indicating that prenuclear events of NF-kappaB activation are not sufficient for Tax-mediated p53 inhibition, but rather NF-kappaB transcriptional activation is critical. Importantly, using phosphospecific antibodies, we demonstrate that phosphorylation of p53 at Ser-15 and Ser-392 correlates with Tax-mediated inhibition. In addition, mutation of p53 at Ser-15 and Ser-392 to alanines renders p53 resistant to Tax inhibition. This report reviews p53 inhibition by Tax and presents our current model. C1 NCI, Basic Res Labs, Virus Tumor Biol Sect, NIH, Bethesda, MD 20892 USA. RP Brady, JN (reprint author), NCI, Basic Res Labs, Virus Tumor Biol Sect, NIH, Bldg 41-B201, Bethesda, MD 20892 USA. NR 46 TC 31 Z9 34 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 NOV 1 PY 2000 VL 16 IS 16 BP 1669 EP 1675 DI 10.1089/08892220050193128 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900015 PM 11080808 ER PT J AU Mahieux, R Pise-Masison, CA Nicot, C Green, P Hall, WW Brady, JN AF Mahieux, R Pise-Masison, CA Nicot, C Green, P Hall, WW Brady, JN TI Inactivation of p53 by HTLV type 1 and HTLV type 2 Tax trans-activators SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID T-CELL LEUKEMIA; VIRUS TYPE-II; TUMOR-SUPPRESSOR; ABERRANT EXPRESSION; GENE PROMOTER; PROTEIN; TRANSACTIVATION; SUBTYPE; PHOSPHORYLATION; IDENTIFICATION AB Human T cell lymphotropic virus type II (HTLV-2) was originally isolated from a patient with a hairy T cell leukemia. It has been associated with rare cases of CD8(+) T lymphoproliferative disorders, and has a controversial role as a pathogen, The loss of p53 function, as a consequence of mutation or inactivation, increases the chances of genetic damage. Indeed, the importance of p53 as a tumor suppressor is evident from the fact that over 60% of all human cancers have a mutant or inactive p53, p53 status has been extensively studied in HTLV-1-infected cell lines. Interestingly, despite the fact that p53 mutations have been found in only a minority of cells, the p53 functions were found to be impaired. We have analyzed the functional activity of the p53 tumor suppressor in cells transformed with HTLV-2 subtypes A and B, As with HTLV-1-infected cells, abundant levels of the p53 protein are detected in HTLV-2 virus-infected cell lines, Using p53 reporter plasmid or induction of p53-responsive genes in response to gamma -irradiation, the p53 was found to be transcriptionally inhibited in HTLV-2-infected cells. Interestingly, although Tax-2A and -2B inactivate p53, the Tax-2A protein appears to inhibit p53 function less efficiently than either Tax-1 or Tax-2B in T cells, but not in fibroblasts. C1 NCI, Basic Res Labs, Sect Anim Models & Retroviral Infect, NIH, Bethesda, MD 20892 USA. Ohio State Univ, Dept Vet Biosci & Mol Virol, Columbus, OH 43210 USA. Ohio State Univ, Dept Immunol & Med Genet, Columbus, OH 43210 USA. Ohio State Univ, Ctr Retrovirus Res, Columbus, OH 43210 USA. Ohio State Univ, Ctr Comprehens Canc, Columbus, OH 43210 USA. Univ Coll Dublin, Dept Microbiol, Dublin 2, Ireland. RP Brady, JN (reprint author), NCI, Basic Res Labs, Sect Anim Models & Retroviral Infect, NIH, Bldg 41-B201, Bethesda, MD 20892 USA. NR 30 TC 21 Z9 21 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 NOV 1 PY 2000 VL 16 IS 16 BP 1677 EP 1681 DI 10.1089/08892220050193137 PG 5 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900016 PM 11080809 ER PT J AU Chun, ACS Zhou, Y Wong, CM Kung, HF Jeang, KT Jin, DY AF Chun, ACS Zhou, Y Wong, CM Kung, HF Jeang, KT Jin, DY TI Coiled-coil motif as a structural basis for the interaction of HTLV type 1 tax with cellular cofactors SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID LEUKEMIA-VIRUS TYPE-1; NF-KAPPA-B; PRIMARY HUMAN-LYMPHOCYTES; I TAX; TRANSACTIVATOR TAX; TRANSFORMING PROTEIN; MEDIATES INTERACTION; BZIP PROTEINS; T-LYMPHOCYTES; DNA-BINDING AB Human T lymphotropic virus type 1 (HTLV-1) Tax is a multifunctional protein centrally involved in transcriptional regulation, cell cycle control, and viral transformation. The regulatory functions of Tax are thought to be mediated through protein-protein interaction with cellular cofactors, Previously we have identified several novel binding partners for Tax, including human mitotic checkpoint protein MAD1 (TXBP181), G-protein pathway suppressor GPS2 (TXBP31), and I kappaB kinase regulatory subunit IKK-gamma, Here we described two additional Tax partners, TXBP151 and TXBP121. A closer examination of the sequences of eight independent cellular Tax-binding proteins identified by us and others revealed that all of them share a single characteristic, a highly structured coiled-coil domain, We also noted that Tax and the Tax-binding coiled-coil proteins can homodimerize. Additionally, the same domain in Tax is responsible for interaction with different coiled-coil proteins, Taken together, our findings point to a particular coiled-coil structure as one of the Tax-recognition motifs, The interaction of Tax with a particular subgroup of cellular coiled-coil proteins represents one mechanism by which Tax dysregulates cell growth and proliferation. C1 Univ Hong Kong, Inst Mol Biol, Hong Kong, Hong Kong, Peoples R China. NIAID, Mol Microbiol Lab, Bethesda, MD 20892 USA. RP Jin, DY (reprint author), Univ Hong Kong, Inst Mol Biol, 8-F Kadoorie Biol Sci Bldg,Pokfulam Rd, Hong Kong, Hong Kong, Peoples R China. RI Jeang, Kuan-Teh/A-2424-2008; Wong, Chi-Ming/A-7627-2013 OI Wong, Chi-Ming/0000-0002-0025-7135 NR 46 TC 29 Z9 29 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 NOV 1 PY 2000 VL 16 IS 16 BP 1689 EP 1694 DI 10.1089/08892220050193155 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900018 PM 11080811 ER PT J AU Kubota, R Nagai, M Kawanishi, T Osame, M Jacobson, S AF Kubota, R Nagai, M Kawanishi, T Osame, M Jacobson, S TI Increased HTLV type 1 Tax-specific CD8(+) cells in HTLV type 1-associated myelopathy/tropical spastic paraparesis: Correlation with HTLV type 1 proviral load SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID I-ASSOCIATED MYELOPATHY; VIRUS TYPE-I; CYTOTOXIC LYMPHOCYTES-T; NEUROLOGICAL DISEASE; PERIPHERAL-BLOOD; CEREBROSPINAL-FLUID; HAM/TSP PATIENTS; SPINAL-CORD; ANTIBODIES; PHENOTYPE AB Less than 1% of individuals infected with the human T lymphotropic virus type 1 (HTLV-1) develop an inflammatory neurological disorder, termed HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), while the vast majority of those infected remain asymptomatic HTLV-1 carriers (ACs). The fundamental viroimmunological differences between these groups are not well understood. To address this issue, we have investigated HTLV-1-specific T cell responses and measured the proviral load in these groups. Frequencies of HTLV-1-specific CD8(+) cells were demonstrated to be significantly higher in HAM/TSP patients than in ACs by using intracellular cytokine staining and soluble divalent HLA-A2/Ig fusion protein loaded with HTLV-1 Tax 11-19 peptide. It is consistent with the observed increase in HTLV-1-specific cytotoxic T lymphocytes in HAM/TSP patients. These CD8(+) cells produced interferon (IFN)-gamma in recognition of HTLV-1 antigens bound to HLAs on the infected CD4(+) cells, Using phenotypic markers indicative for T cell differentiation, memory and/or effector HTLV-1 Tax-specific CD8(+) cells were found to be increased in HLA-A2 HAM/TSP patients. HTLV-1 proviral load was elevated in HAM/TSP patients when compared to ACs, In addition, the proviral load in HAM/TSP patients correlated with the frequency of HTLV-1-specific IFN-gamma (+) CD8(+) cells or Tax-HLA-A2/Ig(+)CD8(+) cells, especially with the effector cells. In contrast, the proviral load inversely correlated with memory cells. These results suggest that HTLV-1 antigens may continuously stimulate HTLV-1-specific CD8(+) cells and differentiate them from memory cells into effector cells irt vivo. These differentiated HTLV-1-specific CD8(+) cells may play a role in the pathogenesis of HAM/TSP. C1 NINDS, Viral Immunol Sect, NIB, NIH, Bethesda, MD 20892 USA. Kagoshima Univ, Sch Med, Dept Internal Med 3, Kagoshima 8908520, Japan. RP Jacobson, S (reprint author), NINDS, Viral Immunol Sect, NIB, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 22 TC 36 Z9 36 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 NOV 1 PY 2000 VL 16 IS 16 BP 1705 EP 1709 DI 10.1089/08892220050193182 PG 5 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900021 PM 11080814 ER PT J AU Azimi, N Mariner, J Jacobson, S Waldmann, TA AF Azimi, N Mariner, J Jacobson, S Waldmann, TA TI How does interleukin 15 contribute to the pathogenesis of HTLV type 1-associated myelopathy/tropical spastic paraparesis? SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID I-ASSOCIATED MYELOPATHY; CYTOTOXIC LYMPHOCYTES-T; LYMPHOTROPIC VIRUS; NEUROLOGICAL DISEASE; CELL LEUKEMIA; BETA-CHAIN; RECEPTOR; IL-15; PROLIFERATION; ACTIVATION AB HTLV-1 is the etiological agent of a neurological disease named HAM/TSP that has clinical characteristics similar to those of multiple sclerosis (MS), The PBMC obtained from HAM/TSP patients undergo spontaneous proliferation in the absence of addition of any exogenous cytokines in an ex vivo culture. This spontaneous proliferation has been thought to be due to the proliferation of T cells. It was demonstrated that this proliferation is, In part, due to the production of IL-2 and its receptor (IL-2R alpha) by HTLV-1-infected T cells. In this review, we demonstrate that IL-15 production also contributes to the spontaneous proliferation of T cells obtained from HAM/TSP PBMC. We provide data indicating that IL-15 expression is elevated in HAM/TSP PBMC when compared to that of the normal donors. Furthermore, we demonstrate that IL-15 overexpression by HTLV-1 is due to Tax trans-activation of its promoter and induction of NF-kappaB transcription factors. On the basis of these studies, we propose a model in which HTLV-1 infection of T cells results in the production of both IL-2 and IL-15 cytokines, growth factors that support the proliferation of T cells. C1 NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. NINDS, Viral Immunol Sect, NIH, Bethesda, MD 20892 USA. RP Azimi, N (reprint author), NCI, Metab Branch, NIH, Bldg 10,Room 4N-102,10 Ctr Dr,MSC 1274, Bethesda, MD 20892 USA. NR 24 TC 20 Z9 21 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 NOV 1 PY 2000 VL 16 IS 16 BP 1717 EP 1722 DI 10.1089/08892220050193209 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900023 PM 11080816 ER PT J AU Albertini, RJ Ardell, SK Judice, SA Jacobson, S Allegretta, M AF Albertini, RJ Ardell, SK Judice, SA Jacobson, S Allegretta, M TI Hypoxanthine-guanine phosphoribosyltransferase reporter gene mutation for analysis of in vivo clonal amplification in patients with HTLV type 1-associated myelopathy/tropical spastic paraparesis SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID FREQUENCY; HUMANS; HPRT AB We tested a surrogate selection approach utilizing mutation at a reporter gene [hypoxanthine-guanine phosphoribosyltransferase (hprt)] as a probe for in vivo cell division, For detection of clonal T cell expansion in human T lymphotropic (HTLV-1) carriers. Peripheral blood samples from HTLV-1-infected individuals with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) were tested to determine the hprt mutant frequency (Mf), Wild-type and hprt mutant T cell clones were isolated, and clonal identity determined by multiplex PCR and DNA sequencing of T cell receptor (TCR) variable region beta -chain (TCR BV) and third complementarity determining regions (CDR3). Seven samples from HAM/TSP patients were tested, and Mfs were within the normal range for adults (mean 11.3 x 10(-6), max 22.4 x 10(-6), min 5.6 x 10(-6)). The frequency of HTLV-1 infection in wild-type and hprt mutant T cells from HAM/TSP patients was determined to identify enrichment in the mutant fraction of cells. This analysis was performed on 196 isolates from 6 individuals with HAM/TSP, In each case, there is enrichment for virally infected cells in the hprt mutant fraction of isolates, Ten mutant and eight wild-type isolates from sample LS42A (Mf 8.4 x 10(-6)) were tested for clonality by TCR BV PCR and sequencing. Of the 10 hprt mutants, there were two in vivo-expanded clones (four isolates with two identical TCRs, or 80% unique TCR sequences). These studies may provide new insights into the precise mechanism of HTLV-1 leukemogenesis, and aid in the study of mutator phenotypes generated by a combination of Tax-mediated in vivo expansion and mutagenesis. C1 BioMosaics Inc, Charlotte, VT USA. Univ Vermont, Genet Toxicol Lab, Burlington, VT USA. NINDS, Viral Immunol Branch, NIH, Bethesda, MD 20205 USA. RP Allegretta, M (reprint author), BioMosaics Inc, 655 Spear St, Burlington, VT 05405 USA. NR 7 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 NOV 1 PY 2000 VL 16 IS 16 BP 1747 EP 1752 DI 10.1089/08892220050193254 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900028 PM 11080821 ER PT J AU D'Agostino, DM Zotti, L Ferro, T Franchini, G Chieco-Bianchi, L Ciminale, V AF D'Agostino, DM Zotti, L Ferro, T Franchini, G Chieco-Bianchi, L Ciminale, V TI The p13(II) protein of HTLV type 1: Comparison with mitochondrial proteins coded by other human viruses SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID BOVINE LEUKEMIA-VIRUS; INFECTED CELL-LINES; MESSENGER-RNAS; I INFECTION; EXPRESSION; APOPTOSIS; REGION; DEATH; PROLIFERATION; INTERLEUKIN-2 AB In addition to the essential regulatory proteins Rex and Tax, the HTLV-1 genome encodes several accessory proteins of yet undefined function, One of these "orphan" proteins, named p13(II), was recently shown to be selectively targeted to mitochondria and to induce specific changes in mitochondrial morphology suggestive of altered inner membrane permeability and swelling. This represented the first report of a retroviral gene product targeted to mitochondria, and suggested that p13(II)-induced alterations in the function of this organelle may play a role in HTLV-1 replication and/or pathogenesis, The more recent findings that both Vpr and Tat of HIV-1 are targeted to mitochondria reinforces the proposed relevance of mitochondrial metabolism to the life cycle of retroviruses, Thus, p13(II), Vpr, and Tat can be added to the growing list of mitochondrial proteins produced by clinically important human viruses, including Epstein-Barr virus, human cytomegalovirus, and hepatitis B virus, Mitochondria are known to play a critical role by providing an amplification loop required for the execution of signaling pathways leading to programmed cell death, The functional consequences of the interactions between viral proteins and mitochondria described so far have been attributed to either the positive or negative? control of apoptotic responses mediated by this organelle, Further analysis of the effects of p13(II) on mitochondrial function is likely to add to our understanding of the mechanisms underlying the development of HTLV-1-associated diseases. C1 Univ Padua, Oncol Sect, Dept Oncol & Surg Sci, I-35128 Padua, Italy. NCI, Basic Res Labs, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Ciminale, V (reprint author), Univ Padua, Oncol Sect, Dept Oncol & Surg Sci, Via Gattamelata 64, I-35128 Padua, Italy. NR 42 TC 19 Z9 19 U1 2 U2 2 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 NOV 1 PY 2000 VL 16 IS 16 BP 1765 EP 1770 DI 10.1089/08892220050193281 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900031 PM 11080824 ER PT J AU Johnson, JM Mulloy, JC Ciminale, V Fullen, J Nicot, C Franchini, G AF Johnson, JM Mulloy, JC Ciminale, V Fullen, J Nicot, C Franchini, G TI The MHC class I heavy chain is a common target of the small proteins encoded by the 3 ' end of HTLV type 1 and HTLV type 2 SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article; Proceedings Paper CT Symposium on HTLV Molecular Biology and Pathogenesis CY MAR 17-19, 2000 CL WARRENTON, VIRGINIA SP NIH ID VIRUS TYPE-II; HAIRY-CELL LEUKEMIA; VACUOLAR H+-ATPASE; READING-FRAME-I; JAK/STAT ACTIVATION; VIRAL REPLICATION; E5 ONCOPROTEIN; P12(I); EXPRESSION; TRANSFORMATION AB Human T cell leukemia/lymphotropic virus types 1 and 2 are two immunologically and phylogenetically related retroviruses that differ in their pathogenicity in vivo. The overall genetic structure of HTLV-1 and -2 is similar. Each contains a unique region at the 3' end of the genome, designated the pX region, p12(I) is a membrane-associated protein encoded by the open reading frame I (ORF I) region of HTLV-1, which lies within the pX region. A corresponding protein, p10(I) is encoded by the ORF I region of HTLV-2 and an additional protein, p11(v), is encoded by ORF V, which overlaps the HTLV-2 ORF I region. As with HTLV-1, the small proteins encoded by the pX region of HTLV-2 appear to be dispensable for viral replication and cellular transformation in vitro. However, the small open reading frames of both viruses are important for viral replication irt vivo, which suggests they may play an important role during the viral life cycle. This study was undertaken to investigate and compare the cellular targets of the p10(I), p11(v), and p12(I) putative proteins. C1 NCI, Basic Res Labs, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. Univ Padua, Dept Oncol & Surg Sci, I-35128 Padua, Italy. RP Franchini, G (reprint author), NCI, Basic Res Labs, 41 Lib Dr,Bldg 41,Room D804,MSC 5055, Bethesda, MD 20892 USA. NR 21 TC 19 Z9 19 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 NOV 1 PY 2000 VL 16 IS 16 BP 1777 EP 1781 DI 10.1089/08892220050193308 PG 5 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 374BQ UT WOS:000165324900033 PM 11080826 ER PT J AU Gea-Banacloche, JC Martino, L Mican, JM Hallahan, CW Baseler, M Stevens, R Lambert, L Polis, M Lane, HC Connors, M AF Gea-Banacloche, JC Martino, L Mican, JM Hallahan, CW Baseler, M Stevens, R Lambert, L Polis, M Lane, HC Connors, M TI Longitudinal changes in CD4(+) T cell antigen receptor diversity and naive/memory cell phenotype during 9 to 26 months of antiretroviral therapy of HIV-infected patients SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; BONE-MARROW TRANSPLANTATION; CLONAL EXPANSIONS; IMMUNE-RESPONSES; REPERTOIRE; ADULTS; NAIVE; REPOPULATION; INDIVIDUALS; KINETICS AB Although skewing of the CD4(+) TCR repertoire in advanced HIV infection is well documented, increases in polyclonality during antiretroviral therapy have been less consistently observed. Ten patients, each with documented abnormalities within the CD4(+) TCR repertoire, were studied by CDR3 spectratyping, semiquantitative PCR, and SSCP during 9-26 months of therapy. Naive and memory cell phenotypes were analyzed by flow cytometry. Six of 10 patients showed increased polyclonality of their TCR repertoires, 1 showed no change, and 3 showed increased TCR skewing, despite suppressed viral replication. Overall, there was no significant change in the percentage of abnormal BV subfamilies (from a mean of 25.5 to 17.1%) or the percentage of naive CD4(+) T cells (from a mean of 18 to 25%). Further, progression of TCR repertoire disruptions was observed in some patients even with suppression of plasma viral RNA below 500 copies/ml. Although a spectrum of changes may be seen within the CD4(+) TCR repertoire in the setting of antiretroviral therapy, increases in polyclonality are observed in some patients. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. SAIC Frederick Canc Res & Dev Ctr, Frederick, MD USA. RP Connors, M (reprint author), NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. OI Polis, Michael/0000-0002-9151-2268 FU NCI NIH HHS [N01-CO-56000] NR 46 TC 18 Z9 18 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 NOV PY 2000 VL 16 IS 17 BP 1877 EP 1886 DI 10.1089/08892220050195838 PG 10 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 375AR UT WOS:000165377800009 PM 11118073 ER PT J AU Blazevic, V Sahgal, N Kessler, HA Landay, AL Shearer, GM AF Blazevic, V Sahgal, N Kessler, HA Landay, AL Shearer, GM TI T cell responses to recall antigens, alloantigen, and mitogen of HIV-infected patients receiving long-term combined antiretroviral therapy SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; LYMPHOCYTE RESPONSES; IMMUNE-RESPONSES; INDIVIDUALS; INHIBITOR; RITONAVIR; CLEARANCE; PROTEASE; VIREMIA; AIDS AB The effect of highly active antiretroviral therapy (HAART) on T cell responses in 30 HIV-infected patients was studied. Lymphocyte proliferation in response to influenza A virus, HIV-1 p24, gp160, allogeneic leukocytes, and mitogen, as well as influenza-specific cytotoxic T lymphocyte (CTL) responses, were measured. AIDS patients had decreased T cell-proliferative responses to influenza and alloantigen compared with asymptomatic patients. Absence of positive proliferative responses of HIV-infected patients to HIV-1 antigens was not associated with increased interleukin 10 production. Correlation was observed between influenza-specific CTL response and T cell proliferation, as well as CD4(+) T lymphocyte counts, indicating the importance of CD4(+) helper T cells for generating antiviral CTL responses. Finally, these results show that HAART-treated asymptomatic patients, but not AIDS patients, have T cell responses comparable to those of control individuals. It remains to be determined whether immune-based therapy will contribute any additional benefit to patients who received HAART. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. Rush Med Coll, Dept Immunol Microbiol, Chicago, IL 60612 USA. Rush Med Coll, Dept Med, Chicago, IL 60612 USA. RP Shearer, GM (reprint author), NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NR 29 TC 13 Z9 14 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 NOV PY 2000 VL 16 IS 17 BP 1887 EP 1893 DI 10.1089/08892220050195847 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 375AR UT WOS:000165377800010 PM 11118074 ER PT J AU Purohit, V AF Purohit, V TI Can alcohol promote aromatization of androgens to estrogens? A review SO ALCOHOL LA English DT Review DE aromatization; androgens; estrogens; alcohol ID HUMAN GRANULOSA-CELLS; BONE-MINERAL DENSITY; POSTMENOPAUSAL WOMEN; AROMATASE-ACTIVITY; STEROID-HORMONES; ETHANOL; CONSUMPTION; ESTRADIOL; INGESTION; EXPOSURE AB Increased aromatization may be a mechanism for feminization of some male alcoholics, as well as for the reported increases in plasma estrogen levels in postmenopausal women subjected to moderate alcohol consumption. Alcohol consumption-related increases in estrogen levels may in turn be partially responsible for the associated decreased risk for coronary artery disease and osteoporosis, as well as for increased risk for breast cancer. The purpose of this review is to evaluate the literature to determine whether alcohol can promote aromatization of androgens to estrogens. In male rats, chronic heavy alcohol administration (36% of total calories = 12- 18 g/kg/day) led to increased aromatization of androgen in the liver, but the results were equivocal for the hypothalamus. In female rats, chronic heavy alcohol administration did not promote aromatization in the hypothalamus exposed to alcohol in utero. In human placental tissue, although ex vivo alcohol administration (less or more than 72 g/day) did not affect the rate of aromatization, in vitro incubation of choriocarcinoma cells with 5-50 mM of alcohol increased estradiol secretion, which could be due to increased aromatization. In in vitro human ovarian granulosa cell studies, alcohol increased, had no effect on, or decreased estradiol secretion, and in one study, 20 mM of alcohol significantly increased aromatization of androstenedione to estrogens. These results may not be fully relevant to normal human ovary because in both studies cells were heavily luteinized by gonadotropins. A study of ovariectomized rats shows that only heavy chronic alcohol intake (4.4 g/kg/day) for 10 weeks can increase plasma estradiol levels and uterine weight, which could be due to increased aromatization or delayed clearance of estradiol. In conclusion, chronic heavy alcohol administration can result in aromatization of androgens in male rat liver. It is not clear whether moderate alcohol intake can produce a similar effect in the liver, nor whether alcohol can potentiate aromatization of androgens in other tissue or organs of male rats. In females, the available information is not adequate to evaluate the effect of alcohol on aromatization. Further studies are required in both genders to evaluate the ability of alcohol (moderate vs. heavy dose) to promote aromatization of androgens to estrogens. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NIAAA, Div Basic Res, Biomed Res Branch, NIH, Bethesda, MD 20892 USA. RP Purohit, V (reprint author), NIAAA, Div Basic Res, Biomed Res Branch, NIH, Suite 402,6000 Execut Blvd, Bethesda, MD 20892 USA. NR 30 TC 47 Z9 48 U1 1 U2 9 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 NOV PY 2000 VL 22 IS 3 BP 123 EP 127 DI 10.1016/S0741-8329(00)00124-5 PG 5 WC Substance Abuse; Pharmacology & Pharmacy; Toxicology SC Substance Abuse; Pharmacology & Pharmacy; Toxicology GA 393WG UT WOS:000166492200002 PM 11163119 ER PT J AU Jeng, CH Chou, JR Hoffer, BJ Wang, Y AF Jeng, CH Chou, JR Hoffer, BJ Wang, Y TI Serotonergic modulation of ethanol-induced electrophysiological depression in young and aged rats SO ALCOHOLISM-CLINICAL AND EXPERIMENTAL RESEARCH LA English DT Article DE ethanol; serotonin; neuromodulation; aging ID CEREBELLAR PURKINJE NEURONS; SPRAGUE-DAWLEY RATS; MESSENGER-RNA; 5-HT2A RECEPTORS; ALCOHOL-DRINKING; RAPHE NUCLEUS; BRAIN-SLICES; DORSAL RAPHE; AUTORECEPTORS; CONSUMPTION AB Background: Ethanol (EtOH)-induced electrophysiological depressions in cerebellar Purkinje neurons have been shown to be potentiated by exogenously applied serotonin (5HT). In this study, we determined whether this modulatory action can be activated by endogenous release from presynaptic serotonergic terminals, and whether such a response is altered by age or rat strain. Methods: Extracellular 5HT levels in cerebellar cortex were measured in real time by in vivo chronoamperometry, by using Nafion-coated carbon fiber electrodes, in anesthetized young (3-5 months old) or aged (18-24 months old) Sprague Dawley and Fischer 344 rats. Some animals were prelesioned with 5,7 dihydroxytryptamine (5,7 DHT). Single unit electrophysiological activity was recorded from cerebellar Purkinje neurons. Serotonin or its presynaptic antagonist methiothepin was applied directly to cerebellar neurons through multibarrel pipettes. Results: Local application of methiothepin dose-dependently induced 5HT overflow in young Sprague Dawley and Fisher 344 rats. Methiothepin-induced 5HT release was decreased significantly in aged or 5,7 DHT-lesioned rats. Local application of methiothepin or 5HT potentiated EtOH-induced electrophysiological depression of Purkinje neurons in young animals of both strains. Methiothepin-potentiated, EtOH-elicited neuronal inhibition was reduced greatly in aged or 5,7 DHT-lesioned rats. Serotonin-facilitated EtOH responses were reduced in the aged Sprague Dawley rats. Conclusions: EtOH-induced electrophysiological responses in cerebellum can be facilitated by endogenous 5HT release by using a 5HT autoreceptor antagonist. Such actions are attenuated in aged rats perhaps through a presynaptic serotonergic mechanism. C1 NIDA, Baltimore, MD 21042 USA. Natl Def Med Ctr, Dept Pharmacol, Taipei, Taiwan. RP Wang, Y (reprint author), NIDA, 5500 Nathan Shock Dr, Baltimore, MD 21042 USA. NR 47 TC 2 Z9 2 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0145-6008 J9 ALCOHOL CLIN EXP RES JI Alcoholism (NY) PD NOV PY 2000 VL 24 IS 11 BP 1730 EP 1741 PG 12 WC Substance Abuse SC Substance Abuse GA 375EA UT WOS:000165385500016 PM 11104121 ER PT J AU Cohen, SG Rizzo, PL AF Cohen, SG Rizzo, PL TI Asthma among the famous - Olive Schreiner (1855-1920) - South African novelist and political activist SO ALLERGY AND ASTHMA PROCEEDINGS LA English DT Biographical-Item C1 NIAID, NIH, Bethesda, MD 20892 USA. RP Cohen, SG (reprint author), NIAID, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU OCEAN SIDE PUBLICATIONS INC PI PROVIDENCE PA 95 PITMAN ST, PROVIDENCE, RI 02906 USA SN 1088-5412 J9 ALLERGY ASTHMA PROC JI Allergy Asthma Proc. PD NOV-DEC PY 2000 VL 21 IS 6 BP 381 EP 396 DI 10.2500/108854100778249132 PG 16 WC Allergy SC Allergy GA 385JW UT WOS:000166001500010 PM 11191106 ER PT J AU Jegalian, K AF Jegalian, K TI The human genome project - Its implications for educators SO AMERICAN BIOLOGY TEACHER LA English DT Editorial Material C1 NHGRI, NIH, Bethesda, MD 20892 USA. RP Jegalian, K (reprint author), NHGRI, NIH, Bethesda, MD 20892 USA. NR 0 TC 5 Z9 5 U1 0 U2 1 PU NATL ASSOC BIOLOGY TEACHERS INC PI RESTON PA 12030 SUNRISE VALLEY DR, #110, RESTON, VA 20191 USA SN 0002-7685 J9 AM BIOL TEACH JI Am. Biol. Teach. PD NOV-DEC PY 2000 VL 62 IS 9 BP 626 EP 627 PG 2 WC Biology; Education, Scientific Disciplines SC Life Sciences & Biomedicine - Other Topics; Education & Educational Research GA 545RG UT WOS:000175230900001 ER PT J AU Garg, R Elam, MB Crouse, JR Davis, KB Kennedy, JW Egan, D Herd, JA Hunninghake, DB Johnson, WC Kostis, JB Sheps, DS Applegate, WB AF Garg, R Elam, MB Crouse, JR Davis, KB Kennedy, JW Egan, D Herd, JA Hunninghake, DB Johnson, WC Kostis, JB Sheps, DS Applegate, WB CA ADMIT investigators TI Effective and safe modification of multiple atherosclerotic risk factors in patients with peripheral arterial disease SO AMERICAN HEART JOURNAL LA English DT Article ID COLESTIPOL-NIACIN THERAPY; NICOTINIC-ACID; HEART-DISEASE; VITAMIN-E; MEN; LOVASTATIN; WARFARIN; HYPERCHOLESTEROLEMIA; PRAVASTATIN; MORTALITY AB Background Patients with peripheral arterial disease (PAD) are at an increased risk of cardiovascular mortality and morbidity and thus are an excellent group in whom to evaluate the feasibility and the effect of an aggressive multifactorial intervention on atherosclerotic vascular disease risk factors. The Arterial Disease Multiple intervention Trial (ADMIT) was designed to determine the efficacy, safety, and compliance of an multifactorial therapy on selected atherosclerotic disease risk factors in patients with PAD. Methods Br a 2 X 2 X 2 factorial design, eligible participants (N = 468) were randomly assigned to low-dose warfarin, antioxidant vitamins, and niacin or its corresponding placebo, and followed up for 1 year. All participants were encouraged to use aspirin. Pravastatin was added to the drug regimen for those who needed to reduce LDI cholesterol to recommended levels. Results Niacin increased HDL cholesterol levels by 30%, with the majority of effect achieved at a dosage of 500 mg twice daily. Warfarin had an anticoagulant effect. The antioxidant vitamins resulted in a significant increase in vitamin E, C, and beta-carotene plasma levels. Overall, compliance was high and few adverse effects were reported. Conclusions ADMIT demonstrates that it is both feasible and safe to modify multiple atherosclerotic disease risk factors effectively with intensive combination therapy in patients with PAD. C1 NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Univ Tennessee, Memphis, TN USA. Wake Forest Univ, Bowman Gray Sch Med, Winston Salem, NC USA. Univ Washington, Seattle, WA 98195 USA. Stat & Epidemiol Res Corp, Seattle, WA USA. Baylor Coll Med, Houston, TX 77030 USA. Univ Minnesota, Minneapolis, MN USA. Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, New Brunswick, NJ USA. Univ N Carolina, Chapel Hill, NC USA. RP Garg, R (reprint author), Eli Lilly & Co, Lilly Corp Ctr, Indianapolis, IN 46225 USA. FU NCRR NIH HHS [MO1RR0021]; NHLBI NIH HHS [N01HC25112, N01HC25116] NR 39 TC 12 Z9 12 U1 1 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 NOV PY 2000 VL 140 IS 5 BP 792 EP 803 DI 10.1067/mhj.2000.110090 PG 12 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 369PB UT WOS:000165075200015 PM 11054628 ER PT J AU Walsh, CR Lloyd-Jones, DM Camargo, CA Giugliano, RP O'Donnell, CJ AF Walsh, CR Lloyd-Jones, DM Camargo, CA Giugliano, RP O'Donnell, CJ TI Clinical trials of unfractionated heparin and low-molecular-weight heparin in addition to aspirin for the treatment of unstable angina pectoris: Do the results apply to all patients? SO AMERICAN JOURNAL OF CARDIOLOGY LA English DT Article ID ACUTE MYOCARDIAL-INFARCTION; CORONARY-ARTERY DISEASE; MANAGEMENT; ISCHEMIA; DEATH AB We sought to determine how the results of available randomized controlled trials of intravenous unfractionated heparin (UH) and low-molecular weight heparin (LMWH) apply to unselected patients with unstable angina pectoris (UAP). Although UH is widely used in addition to aspirin for treatment of UAP, the evidence is weak for a net benefit over aspirin alone. LMWH preparations may confer a net benefit over UH for the treatment of UAP in clinical trials. It is not clear, however, how trial results are generalized to unselected patients with UAP, Using criteria from the Agency for Health Care Policy and Research Unstable Angina Clinical Practice Guideline, we identified 277 consecutive patients with primary UAP, Exclusion criteria were applied from 6 trials of UH in addition to aspirin and 5 trials of LMWH in addition to aspirin for the treatment of UAP. Clinical outcomes were compared among ineligible and eligible patients for trial enrollment. Patients meeting exclusion criteria were older and had more extensive coexisting medical illness than eligible patients for trial enrollment. Thirty-eight percent to 42% of our study population met greater than or equal to1 exclusion criteria for each of the 6 trials of UH, and 14% to 46% met greater than or equal to1 exclusion criteria for each of the 5 LMWH trials. The I-year all-cause death rate was higher in UH ineligible patients compared with UH eligible patients (16% vs 4%, p = 0.003) and in LMWH ineligible patients compared with LMWH eligible patients (16% vs 7%, p = 0.005). Thus, clinical trials of UH and LMWH may have limited generalizability to unselected patients with UAP, many of whom have characteristics that would exclude them from trial enrollment and put them at risk for adverse outcomes. (C)2000 by Excerpta Medica, Inc. C1 NHLBI Framingham Heart Study, Framingham, MA 01702 USA. Massachusetts Gen Hosp, Dept Med, Div Cardiol, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Emergency Med, Boston, MA 02114 USA. Harvard Univ, Sch Med, Channing Lab, Boston, MA 02115 USA. Harvard Univ, Brigham & Womens Hosp, Sch Med, Dept Med,Cardiovasc Div, Boston, MA 02115 USA. RP O'Donnell, CJ (reprint author), NHLBI Framingham Heart Study, 5 Thurber St, Framingham, MA 01702 USA. RI Lloyd-Jones, Donald/C-5899-2009 FU NHLBI NIH HHS [HL-03533, HL-07575] NR 21 TC 5 Z9 5 U1 0 U2 3 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 NOV 1 PY 2000 VL 86 IS 9 BP 908 EP 912 DI 10.1016/S0002-9149(00)01120-6 PG 5 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 369YU UT WOS:000165096000003 PM 11053697 ER PT J AU Hubbard, VS AF Hubbard, VS TI Defining overweight and obesity: what are the issues? SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Editorial Material C1 NIH, Div Nutr Res Coordinat, Bethesda, MD 20892 USA. NIDDKD, Nutrit Sci Branch, Bethesda, MD 20892 USA. RP Hubbard, VS (reprint author), NIH, Div Nutr Res Coordinat, Rockledge 1,Suite 8048,6705 Rockledge Dr,MSC 7973, Bethesda, MD 20892 USA. NR 8 TC 26 Z9 27 U1 0 U2 1 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 NOV PY 2000 VL 72 IS 5 BP 1067 EP 1068 PG 2 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 369XN UT WOS:000165093200001 PM 11063427 ER PT J AU Lauer, RM Obarzanek, E Hunsberger, SA Van Horn, L Hartmuller, VW Barton, BA Stevens, VJ Kwiterovich, PO Franklin, FA Kimm, SYS Lasser, NL Simons-Morton, DG AF Lauer, RM Obarzanek, E Hunsberger, SA Van Horn, L Hartmuller, VW Barton, BA Stevens, VJ Kwiterovich, PO Franklin, FA Kimm, SYS Lasser, NL Simons-Morton, DG TI Efficacy and safety of lowering dietary intake of total fat, saturated fat, and cholesterol in children with elevated LDL cholesterol: the Dietary Intervention Study in Children SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Article; Proceedings Paper CT International Colloquium on Fat Intake During Childhood CY JUN 08-09, 1998 CL HOUSTON, TEXAS SP Mars Inc DE efficacy; safety; lower-fat diets; prepubertal children; Dietary Intervention Study in Children; dietary fat; LDL cholesterol ID DENSITY-LIPOPROTEIN CHOLESTEROL; SERUM-LIPIDS; GROWTH; DISC; STANDARDIZATION; HYPERLIPIDEMIA; ADOLESCENTS; MATURATION; INFANCY; DISEASE AB Background: Few studies have shown the efficacy and safety of lower-fat diets in children. Objective: Our objective was to assess the efficacy and safety of lowering dietary intake of total fat, saturated fat, and cholesterol to decrease LDL-cholesterol concentrations in children. Design: A 6-center, randomized controlled clinical trial was carried out in 663 children aged 8-10 y with LDL-cholesterol concentrations greater than the 80th and less than the 98th percentiles for age and sex. The children were randomly assigned to either an intervention group or a usual care group. Behavioral intervention promoted adherence to a diet providing 28% of energy from total fat, < 8% from saturated fat, 9% from polyunsaturated fat, and < 0.018 mg cholesterol(.)kJ(-1.)d(-1) (not to exceed 150 mg/d). The primary efficacy measure was mean LDL cholesterol and the safety measures were mean height and serum ferritin concentration at 3 y. Results: At 3 y, dietary total fat, saturated fat, and cholesterol were lower in the intervention group than in the usual care group tall P < 0.001). LDL cholesterol decreased in the intervention and usual care groups by 0.40 mmol/L (15.4 mg/dL) and 0.31 mmol/L (11.9 mg/dL), respectively. With adjustment for baseline concentration, sex, and missing data, the mean difference between groups was -0.08 mmol/L (95% CI: -0.15, -0.01), or -3.23 mg/dL (95% CI: -5.6, -0.5) (P = 0.016). There were no significant differences between groups in adjusted mean height or serum ferritin. Conclusion: Dietary changes are effective in achieving modest lowering of LDL cholesterol over 3 y while maintaining adequate growth, iron stores, nutritional adequacy, and psychological well-being during the critical growth period of adolescence. C1 Maryland Med Res Inst, DISC Coordinating Ctr, Baltimore, MD 21210 USA. Univ Iowa Hosp & Clin, Dept Pediat, Iowa City, IA 52242 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Northwestern Univ, Sch Med, Dept Prevent Med, Chicago, IL USA. Johns Hopkins Hosp, Sch Med, Dept Pediat, Baltimore, MD 21287 USA. Kaiser Fdn Hosp, Kaiser Permanente Ctr Hlth Res, Portland, OR USA. Johns Hopkins Hosp, Childrens Med & Surg Ctr, Baltimore, MD 21287 USA. Childrens Hosp Alabama, Dept Gastrointestinal Nutr, Birmingham, AL USA. Univ Pittsburgh, Sch Med, Dept Clin Epidemiol & Prevent Med, Pittsburgh, PA 15261 USA. New Jersey Med Sch, Prevent Cardiol Program, Newark, NJ USA. RP Lauer, RM (reprint author), Maryland Med Res Inst, DISC Coordinating Ctr, 600 Wyndhurst Ave, Baltimore, MD 21210 USA. NR 59 TC 28 Z9 29 U1 0 U2 2 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 NOV PY 2000 VL 72 IS 5 SU S BP 1332S EP 1342S PG 11 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 371KE UT WOS:000165177500011 PM 11063475 ER PT J AU Troiano, RP Briefel, RR Carroll, MD Bialostosky, K AF Troiano, RP Briefel, RR Carroll, MD Bialostosky, K TI Energy and fat intakes of children and adolescents in the United States: data from the National Health and Nutrition Examination Surveys SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Article; Proceedings Paper CT International Colloquium on Fat Intake During Childhood CY JUN 08-09, 1998 CL HOUSTON, TEXAS SP Mars Inc DE National Health and Nutrition Examination; Survey; NHANES; energy intake; dietary fats; beverages; health surveys; nutrition surveys; trends; child; preschool; adolescence; United States ID RISK-FACTORS; OBESITY; TRENDS; EPIDEMIOLOGY; OVERWEIGHT AB Background: Dietary factors related to body weight and chronic disease risk are of interest because of recent increases in the prevalence of overweight. Objective: Secular trends in energy and fat intakes of youths aged 2-19 y were assessed. Current intakes were compared with recommendations. Design: Dietary 24-h recall data from the third National Health and Nutrition Examination Survey (1988-1994) and earlier national surveys were examined. Results: Mean energy intake changed little from the 1970s to 1988-1994 except for an increase among adolescent females. Over the same time period, the mean percentage of energy from total and saturated fat decreased, but remained above recommendations, with overall means of 33.5% of energy from fat and 12.2% of energy from saturated fat, in 1988-1994, approximate to1 in 4 youths met the recommendations for intakes of fat and saturated fat and 3 in 4 met the recommendation for cholesterol intake. Beverages contributed 20-24% of energy across all ages and soft drinks provided 8% of energy in adolescents. Except for adolescent girls, beverage energy contributions were generally higher among overweight than nonoverweight youths; soft drink energy contribution was higher among overweight youths than among nonoverweight youths for all groups. Conclusions: The lack of evidence of a general increase in energy intake among youths despite an increase in the prevalence of overweight suggests that physical inactivity is a major public health challenge in this age group. Efforts to increase physical activity and decrease nonnutritive sources of energy may be important approaches to counter the rise in overweight prevalence. C1 NCI, Div Canc Control & Populat Sci, NIH, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Natl Ctr Hlth Stat, Div Hlth Examinat Stat, Hyattsville, MD 20782 USA. RP Troiano, RP (reprint author), NCI, Div Canc Control & Populat Sci, NIH, ARP Execut Plaza N,Room 4005,6130 Execut Blvd, Bethesda, MD 20892 USA. NR 54 TC 181 Z9 191 U1 0 U2 15 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 NOV PY 2000 VL 72 IS 5 SU S BP 1343S EP 1353S PG 11 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 371KE UT WOS:000165177500012 PM 11063476 ER PT J AU Alavanja, MCR Lubin, JH Mahaffey, JA Brownson, RC AF Alavanja, MCR Lubin, JH Mahaffey, JA Brownson, RC TI Re: "Residential radon gas exposure and lung cancer: The Iowa Radon Lung Cancer Study" SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Letter ID INDOOR RADON; RISK C1 NCI, Div Canc Epidemiol & Genet, Rockville, MD 20892 USA. Battelle Mem Inst, Richland, WA 99352 USA. St Louis Univ, Sch Publ Hlth, Dept Community Med, St Louis, MO 63108 USA. RP Alavanja, MCR (reprint author), NCI, Div Canc Epidemiol & Genet, Rockville, MD 20892 USA. NR 8 TC 6 Z9 7 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 NOV 1 PY 2000 VL 152 IS 9 BP 895 EP 895 DI 10.1093/aje/152.9.895 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 369ZN UT WOS:000165097800016 PM 11085403 ER PT J AU Ghosh, S Watanabe, RM Valle, TT Hauser, ER Magnuson, VL Langefeld, CD Ally, DS Mohlke, KL Silander, K Kohtamaki, K Chines, P Balow, J Birznieks, G Chang, J Eldridge, W Erdos, MR Karanjawala, ZE Knapp, JI Kudelko, K Martin, C Morales-Mena, A Musick, A Musick, T Pfahl, C Porter, R Rayman, JB Rha, D Segal, L Shapiro, S Sharaf, R Shurtleff, B So, A Tannenbaum, J Te, C Tovar, J Unni, A Welch, C Whiten, R Witt, A Blaschak-Harvan, J Douglas, JA Duren, WL Epstein, MP Fingerlin, TE Kaleta, HS Lange, EM Li, C McEachin, RC Stringham, HM Trager, E White, PP Eriksson, J Toivanen, L Vidgren, G Nylund, SJ Tuomilehto-Wolf, E Ross, EH Demirchyan, E Hagopian, WA Buchanan, TA Tuomilehto, J Bergman, RN Collins, FS Boehnke, M AF Ghosh, S Watanabe, RM Valle, TT Hauser, ER Magnuson, VL Langefeld, CD Ally, DS Mohlke, KL Silander, K Kohtamaki, K Chines, P Balow, J Birznieks, G Chang, J Eldridge, W Erdos, MR Karanjawala, ZE Knapp, JI Kudelko, K Martin, C Morales-Mena, A Musick, A Musick, T Pfahl, C Porter, R Rayman, JB Rha, D Segal, L Shapiro, S Sharaf, R Shurtleff, B So, A Tannenbaum, J Te, C Tovar, J Unni, A Welch, C Whiten, R Witt, A Blaschak-Harvan, J Douglas, JA Duren, WL Epstein, MP Fingerlin, TE Kaleta, HS Lange, EM Li, C McEachin, RC Stringham, HM Trager, E White, PP Eriksson, J Toivanen, L Vidgren, G Nylund, SJ Tuomilehto-Wolf, E Ross, EH Demirchyan, E Hagopian, WA Buchanan, TA Tuomilehto, J Bergman, RN Collins, FS Boehnke, M TI The Finland-United States Investigation of Non-Insulin-Dependent Diabetes Mellitus Genetics (FUSION) study. I. An autosomal genome scan for genes that predispose to type 2 diabetes SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID SIB-PAIR LINKAGE; MAJOR SUSCEPTIBILITY LOCUS; PIMA-INDIANS; MEXICAN-AMERICANS; COMPLEX TRAITS; MAPPING GENES; WIDE SEARCH; ONSET; NIDDM; MAPS AB We performed a genome scan at an average resolution of 8 cM in 719 Finnish sib pairs with type 2 diabetes. Our strongest results are for chromosome 20, where we observe a weighted maximum LOD score (MLS) of 2.15 at map position 69.5 cM from pter and secondary weighted LOD-score peaks of 2.04 at 56.5 cM and 1.99 at 17.5 cM. Our next largest MLS is for chromosome 11 (MLS = 1.75 at 84.0 cM), followed by chromosomes 2 (MLS = 0.87 at 5.5 cM), 10 (MLS = 0.77 at 75.0 cM), and 6 (MLS = 0.61 at 112.5 cM), all under an additive model. When we condition on chromosome 2 at 8.5 cM, the MLS for chromosome 20 increases to 5.50 at 69.0 cM (P = .0014). An ordered-subsets analysis based on families with high or low diabetes-related quantitative traits yielded results that support the possible existence of disease-predisposing genes on chromosomes 6 and 10. Genomewide linkage-disequilibrium analysis using microsatellite marker data revealed strong evidence of association for D22S423 (P = .00007). Further analyses are being carried out to confirm and to refine the location of these putative diabetes-predisposing genes. C1 Univ Michigan, Sch Publ Hlth, Dept Biostat, Ann Arbor, MI 48109 USA. NHGRI, Genet & Mol Biol Branch, Bethesda, MD 20892 USA. Natl Publ Hlth Inst, Dept Epidemiol & Hlth Promot, Diabet & Genet Epidmiol Unit, Helsinki, Finland. Univ So Calif, Keck Sch Med, Dept Physiol & Biophys, Los Angeles, CA USA. Univ So Calif, Keck Sch Med, Dept Med, Los Angeles, CA USA. Univ Washington, Dept Med, Seattle, WA USA. RP Boehnke, M (reprint author), Univ Michigan, Sch Publ Hlth, Dept Biostat, 1420 Washington Hts, Ann Arbor, MI 48109 USA. RI Addington, Anjene/C-3460-2008; Li, Chun/B-8388-2012 FU NHGRI NIH HHS [HG00376, R01 HG000376, T32 HG000040, Z01 HG000040]; NIDDK NIH HHS [DK09525, DK27619, F32 DK009525, R01 DK027619, R01 DK029867, R37 DK027619] NR 56 TC 193 Z9 198 U1 0 U2 7 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD NOV PY 2000 VL 67 IS 5 BP 1174 EP 1185 PG 12 WC Genetics & Heredity SC Genetics & Heredity GA 369WW UT WOS:000165091600015 PM 11032783 ER PT J AU Watanabe, RM Ghosh, S Langefeld, CD Valle, TT Hauser, ER Magnuson, VL Mohlke, KL Silander, K Ally, DS Chines, P Blaschak-Harvan, J Douglas, JA Duren, WL Epstein, MP Fingerlin, TE Kaleta, HS Lange, EM Li, C McEachin, RC Stringham, HM Trager, E White, PP Balow, J Birznieks, G Chang, J Eldridge, W Erdos, MR Karanjawala, ZE Knapp, JI Kudelko, K Martin, C Morales-Mena, A Musick, A Musick, T Pfahl, C Porter, R Rayman, JB Rha, D Segal, L Shapiro, S Sharaf, R Shurtleff, B So, A Tannenbaum, J Te, C Tovar, J Unni, A Welch, C Whiten, R Witt, A Kohtamaki, K Ehnholm, C Eriksson, J Toivanen, L Vidgren, G Nylund, SJ Tuomilehto-Wolf, E Ross, EH Demirchyan, E Hagopian, WA Buchanan, TA Tuomilehto, J Bergman, RN Collins, FS Boehnke, M AF Watanabe, RM Ghosh, S Langefeld, CD Valle, TT Hauser, ER Magnuson, VL Mohlke, KL Silander, K Ally, DS Chines, P Blaschak-Harvan, J Douglas, JA Duren, WL Epstein, MP Fingerlin, TE Kaleta, HS Lange, EM Li, C McEachin, RC Stringham, HM Trager, E White, PP Balow, J Birznieks, G Chang, J Eldridge, W Erdos, MR Karanjawala, ZE Knapp, JI Kudelko, K Martin, C Morales-Mena, A Musick, A Musick, T Pfahl, C Porter, R Rayman, JB Rha, D Segal, L Shapiro, S Sharaf, R Shurtleff, B So, A Tannenbaum, J Te, C Tovar, J Unni, A Welch, C Whiten, R Witt, A Kohtamaki, K Ehnholm, C Eriksson, J Toivanen, L Vidgren, G Nylund, SJ Tuomilehto-Wolf, E Ross, EH Demirchyan, E Hagopian, WA Buchanan, TA Tuomilehto, J Bergman, RN Collins, FS Boehnke, M TI The Finland-United States Investigation of Non-Insulin-Dependent Diabetes Mellitus Genetics (FUSION) study. II. An autosomal genome scan for diabetes-related quantitative-trait loci SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID BODY-MASS INDEX; PIMA-INDIANS; GLUCOSE-TOLERANCE; LINKAGE ANALYSIS; SUSCEPTIBILITY GENES; MEXICAN-AMERICANS; FINNISH FAMILIES; MINIMAL-MODEL; MAPPING GENES; WIDE SEARCH AB Type 2 diabetes mellitus is a complex disorder encompassing multiple metabolic defects. We report results from an autosomal genome scan for type 2 diabetes-related quantitative traits in 580 Finnish families ascertained for an affected sibling pair and analyzed by the variance components-based quantitative-trait locus (QTL) linkage approach. We analyzed diabetic and nondiabetic subjects separately, because of the possible impact of disease on the traits of interest. In diabetic individuals, our strongest results were observed on chromosomes 3 (fasting C-peptide/glucose: maximum LOD score [MLS] = 3.13 at 53.0 cM) and 13 (body-mass index: MLS = 3.28 at 5.0 cM). In nondiabetic individuals, the strongest results were observed on chromosomes 10 (acute insulin response: MLS = 3.11 at 21.0 cM), 13 (2-h insulin: MLS = 2.86 at 65.5 cM), and 17 (fasting insulin/glucose ratio: MLS = 3.20 at 9.0 cM). In several cases, there was evidence for overlapping signals between diabetic and nondiabetic individuals; therefore we performed joint analyses. In these joint analyses, we observed strong signals for chromosomes 3 (body-mass index: MLS = 3.43 at 59.5 cM), 17 (empirical insulin-resistance index: MLS = 3.61 at 0.0 cM), and 19 (empirical insulin-resistance index: MLS = 2.80 at 74.5 cM). Integrating genome-scan results from the companion article by Ghosh et al., we identify several regions that may harbor susceptibility genes for type 2 diabetes in the Finnish population. C1 Univ Michigan, Sch Publ Hlth, Dept Biostat, Ann Arbor, MI 48109 USA. NHGRI, Genet & Mol Biol Branch, Bethesda, MD 20892 USA. Natl Publ Hlth Inst, Dept Epidemiol & Hlth Promot, Diabet & Genet Epidemiol Unit, Helsinki, Finland. Univ So Calif, Keck Sch Med, Dept Physiol & Biophys, Los Angeles, CA USA. Univ So Calif, Keck Sch Med, Dept Med, Los Angeles, CA USA. Univ Washington, Dept Med, Seattle, WA USA. RP Boehnke, M (reprint author), Univ Michigan, Sch Publ Hlth, Dept Biostat, 1420 Washington Hts, Ann Arbor, MI 48109 USA. RI Addington, Anjene/C-3460-2008; Li, Chun/B-8388-2012 FU NHGRI NIH HHS [HG00376, T32 HG000040, Z01 HG000040, R01 HG000376]; NIDDK NIH HHS [F32 DK009525, R01 DK027619, R01 DK029867, R37 DK027619] NR 54 TC 122 Z9 124 U1 0 U2 3 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD NOV PY 2000 VL 67 IS 5 BP 1186 EP 1200 PG 15 WC Genetics & Heredity SC Genetics & Heredity GA 369WW UT WOS:000165091600016 PM 11032784 ER PT J AU Jiao, XD Munier, FL Iwata, F Hayakawa, M Kanai, A Lee, J Schorderet, DF Chen, MS Kaiser-Kupfer, M Hejtmancik, JF AF Jiao, XD Munier, FL Iwata, F Hayakawa, M Kanai, A Lee, J Schorderet, DF Chen, MS Kaiser-Kupfer, M Hejtmancik, JF TI Genetic linkage of Bietti crystallin corneoretinal dystrophy to chromosome 4q35 SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article AB Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessive retinal degeneration characterized by multiple glistening intraretinal dots scattered over the fundus, degeneration of the retina, and sclerosis of the choroidal vessels, ultimately resulting in progressive night blindness and constriction of the visual field. Although BCD has been associated with abnormalities in fatty-acid metabolism and absence of fatty-acid binding by two cytosolic proteins, the genetic basis of BCD is unknown. We report linkage of the BCD locus to D4S426 (maximum LOD score [Z(max)] 4.81; recombination fraction [theta] 0), D4S2688 (Z(max) = 3.97; theta = 0), and D4S2299 (Z(max) = 5.31; theta = 0), on chromosome 4q35-4qtel. Multipoint analysis confirmed linkage to the region telomeric of D4S1652 with a Z(max) of 5.3 located 4 cM telomeric of marker D4S2930. C1 NEI, OGCSB, NIH, Bethesda, MD 20892 USA. Juntendo Univ, Sch Med, Tokyo 113, Japan. Jules Gonin Eye Hosp, Oculogenet Unit, Lausanne, Switzerland. Div Med Genet, Lausanne, Switzerland. Natl Taiwan Univ Hosp, Dept Ophthalmol, Taipei, Taiwan. RP Hejtmancik, JF (reprint author), NEI, OGCSB, NIH, Bldg 10,Room 10B10,10 Ctr Dr,MSC 1860, Bethesda, MD 20892 USA. NR 9 TC 43 Z9 47 U1 1 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD NOV PY 2000 VL 67 IS 5 BP 1309 EP 1313 PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 369WW UT WOS:000165091600027 PM 11001583 ER PT J AU Okechukwu, CN Orzol, SM Held, PJ Pereira, BJG Agodoa, LYC Wolfe, RA Port, FK AF Okechukwu, CN Orzol, SM Held, PJ Pereira, BJG Agodoa, LYC Wolfe, RA Port, FK TI Characteristics and treatment of patients not reusing dialyzers in reuse units SO AMERICAN JOURNAL OF KIDNEY DISEASES LA English DT Article; Proceedings Paper CT 31st Congress of the American-Society-of-Nephrology CY OCT 24-28, 1998 CL PHILADELPHIA, PENNSYLVANIA SP Amer Soc Nephrol DE hemodialysis (HD); dialyzer reuse; Kt/V; US Renal Data System (USRDS); dialysis prescription; dialysis membrane ID DIALYSIS; HEMODIALYSIS; SURVIVAL; MORTALITY; SAFETY AB Dialyzer reuse is practiced in more than 75% of the patients and dialysis units in the United States, However, reuse is not practiced in a small fraction of patients treated in reuse units (RUUs). This study evaluates both patient and facility characteristics associated with nonreuse in RUUs, The data source is from the Dialysis Mortality and Morbidity Study, Waves 1, 3, and 4, of the US Renal Data System. Only facilities that practiced dialyzer reuse were included in the analysis. A total of 12,094 patients from 1,095 reuse facilities were studied. Patients undergoing hemodialysis as of December 31, 1993, were selected. Of all patients treated in RUUs, 8% did not reuse dialyzers. Nonreuse was significantly (P < 0.02) more common, based on adjusted odds ratios (ORs), among patients who were younger (OR = 1.16 per 10 years younger), had primary glomerulonephritis (OR = 1.26 versus diabetes), had lower serum albumin level (OR = 1.72 per 1 g/dL lower), had more years on dialysis, and had higher level of education. Nonreuse patients were more likely to be treated with low-flux dialyzers (OR = 7.35; P < 0.0001) and have a lower dialysis dose, No reuse was more likely in larger units and in not-for-profit and hospital-based units. Patient refusal accounted for one fourth of nonreuse in RUUs and was associated with the same factors, as well as with fewer comorbid conditions and non-Hispanic ethnicity. Significant geographic variations (up to eightfold) were documented. Nonreuse patients are treated with smaller, low-flux dialyzers and, on average, receive a lower Kt/V than reuse patients in the same units, (C) 2000 by the National Kidney Foundation, Inc. C1 Univ Michigan, Dept Internal Med, Kidney Epidemiol & Cost Ctr, US Renal Data Syst, Ann Arbor, MI 48109 USA. Univ Michigan, Dept Epidemiol, Kidney Epidemiol & Cost Ctr, US Renal Data Syst, Ann Arbor, MI 48109 USA. Univ Michigan, Dept Biostat, Kidney Epidemiol & Cost Ctr, US Renal Data Syst, Ann Arbor, MI 48109 USA. Univ Renal Res & Educ Associat, Ann Arbor, MI USA. NIDDKD, NIH, Bethesda, MD 20892 USA. New England Med Ctr, Boston, MA 02111 USA. RP Port, FK (reprint author), 315 W Huron St,Ste 240, Ann Arbor, MI 48103 USA. NR 24 TC 6 Z9 7 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 NOV PY 2000 VL 36 IS 5 BP 991 EP 999 DI 10.1053/ajkd.2000.19101 PG 9 WC Urology & Nephrology SC Urology & Nephrology GA 368GJ UT WOS:000090109100014 PM 11054356 ER PT J AU Wolfe, RA Ashby, VB Milford, EL Bloembergen, WE Agodoa, LYC Held, PJ Port, FK AF Wolfe, RA Ashby, VB Milford, EL Bloembergen, WE Agodoa, LYC Held, PJ Port, FK TI Differences in access to cadaveric renal transplantation in the United States SO AMERICAN JOURNAL OF KIDNEY DISEASES LA English DT Article DE diabetic end-stage renal disease (ESRD); dialysis; end-stage renal disease (ESRD); gender; race; cadaveric renal transplantation; transplant, patient selection; transplant, waiting list ID RACIAL-DIFFERENCES; RACE; BLACKS; GENDER; IMPACT; SEX AB This national study compares waitlisting and transplantation rates by gender, race, and diabetes and evaluates physiologic factors (panel-reactive antibodies [PRA], blood type, HLA matchability) and related practices (early and multiple waitlisting) as explanatory factors. This longitudinal study of the time to transplant waitlisting among 228,552 incident end-stage renal disease (ESRD) dialysis patients and to cadaveric transplantation among 46,164 waitlist dialysis patients (n = 23,275 first cadaveric transplants) used US data for 1991 to 1997. Relative rates of waitlisting (RRWL) after ESRD onset and of cadaveric transplantation (RRTx) after waitlist (Cox proportional hazards models) were adjusted for age, race, sex, ESRD cause, region, and incidence/waitlist year. We found that women have an RRWL = 0.84 (P < 0.0001) and RRTx = 0.86 (P < 0.0001). PRA levels can explain the difference in the transplantation rate, because accounting for PRA gives an adjusted RRTx = 0.98 (NS) for women. For blacks versus whites, the RRWL = 0.59 (P < 0.0001) and RRTx = 0.55 (P < 0.0001). However, the transplantation rate can only partly be explained by ABO types, rare HLA types, and early and multiple waitlisting (adjusted RRTx = 0.67 [P < 0.0001]). For diabetes versus glomerulonephritis, the RRWL = 0.52 (P < 0.0001) and RRTx = 0.98 (NS). Older patients (40 to 59 years of age) are less likely to be waitlisted and to receive a transplant after waitlisting (RRWL = 0.57 [P < 0.0001], RRTx = 0.88 [P < 0.0001]) versus younger patients (ages 18 to 39 years). These results indicate substantial differences by age, sex, race, and diabetes in rates of waitlisting for transplantation and by age and race for transplantation after waitlisting. These differences by race were not explained by referral practices or the physiologic factors studied here. (C) 2000 by the National Kidney Foundation, Inc. C1 Univ Michigan, Ann Arbor, MI 48103 USA. Brigham & Womens Hosp, Boston, MA 02115 USA. NIDDKD, Bethesda, MD 20892 USA. Univ Renal Res Educ Assoc, Ann Arbor, MI USA. RP Wolfe, RA (reprint author), Univ Michigan, 315 W Huron,Ste 240, Ann Arbor, MI 48103 USA. FU NIDDK NIH HHS [N01-DK-3-22-2] NR 27 TC 115 Z9 116 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 NOV PY 2000 VL 36 IS 5 BP 1025 EP 1033 DI 10.1053/ajkd.2000.19106 PG 9 WC Urology & Nephrology SC Urology & Nephrology GA 368GJ UT WOS:000090109100019 PM 11054361 ER PT J AU Romero, R Maymon, E Pacora, P Gomez, R Mazor, M Yoon, BH Berry, SM AF Romero, R Maymon, E Pacora, P Gomez, R Mazor, M Yoon, BH Berry, SM TI Further observations on the fetal inflammatory response syndrome: A potential homeostatic role for the soluble receptors of tumor necrosis factor alpha SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article; Proceedings Paper CT 20th Annual Meeting of the Society-for-Maternal-Fetal-Medicine CY JAN 31-FEB 05, 2000 CL MIAMI BEACH, FLORIDA SP Soc Maternal Fetal Med DE tumor necrosis factor alpha; tumor necrosis factor alpha receptors; preterm premature rupture of the membranes; preterm labor ID SEPTIC SHOCK; TNF RECEPTOR; PRETERM PARTURITION; NEONATAL SEPSIS; FUSION PROTEIN; MICE; PATHOPHYSIOLOGY; INTERLEUKIN-6; INFECTION; LIGAND AB OBJECTIVE: The fetal inflammatory response syndrome is a subclinical condition frequently present in preterm labor and preterm premature rupture of the membranes and is associated with increased perinatal morbidity and mortality. Tumor necrosis factor a is a mediator of septic shock and death, and it exerts its biologic effects by interacting with 2 receptors, TNF-R1 and TNF-R2. Soluble tumor necrosis factor receptors can buffer the biologic activity and protect against the deleterious effects of tumor necrosis factor alpha. The purpose of this study was to determine the behavior of soluble tumor necrosis factor receptors in fetuses with and without fetal inflammatory response syndrome. STUDY DESIGN: Fetal blood sampling was performed in patients with preterm labor (n = 95) and preterm premature rupture of the membranes (n = 39). Control samples were obtained from fetuses who were undergoing blood sampling for clinical indications and had normal outcomes (n = 21). Fetal inflammatory response syndrome was defined as a fetal plasma interleukin 6 concentration >11 pg/mL. Concentrations of interleukin 6 and TNF-R1 and TNF-R2 were determined by use of sensitive and specific immunoassays. Analysis of covariance was used for statistical analysis. RESULTS: (1)TNF-R1 and TNF-R2 were detectable in all samples, and their concentrations decreased with advancing gestational age (r = -0.8 and r = -0.7; P < .0001 and P < .001, respectively). (2) The mean fetal plasma concentrations of TNF-RI and TNF-R2 were significantly higher in fetuses with fetal inflammatory response syndrome than in those without the syndrome after adjustment for gestational age and fetal membrane status (TNF-R1: no fetal inflammatory response syndrome, mean +/- SE. 3473.7 +/- 128.8 pg/mL; vs fetal inflammatory response syndrome, mean +/- SE, 4079.9 +/- 190.7 pg/mL; P < .005;TNF-R2: no fetal inflammatory response syndrome, mean +/- SE, 6033.2 +/- 235.4 pg/mL; vs fetal inflammatory response syndrome, mean +/- SE, 7783.1 +/- 342.8 pg/mL; P < .0001). (3) Fetuses of patients who delivered within 72 hours of cordocentesis had significantly higher concentrations of TNF-R1 and TNF-R2 receptors than those with longer latency periods (P < .05 for each). CONCLUSION: The fetal inflammatory response syndrome is associated with increased availability of the soluble receptors of tumor necrosis factor in fetal plasma. These factors may attenuate the deleterious effects of tumor necrosis factor alpha. C1 Wayne State Univ, Hutzel Hosp, NICHHD, Perinatol Res Branch,Dept Obstet & Gynecol, Detroit, MI 48201 USA. Ben Gurion Univ Negev, Dept Obstet & Gynecol, IL-84105 Beer Sheva, Israel. Seoul Natl Univ, Dept Obstet & Gynecol, Seoul 151, South Korea. RP Romero, R (reprint author), Wayne State Univ, Hutzel Hosp, NICHHD, Perinatol Res Branch,Dept Obstet & Gynecol, 4707 St Antoine Blvd, Detroit, MI 48201 USA. RI Yoon, Bo Hyun/H-6344-2011 NR 30 TC 44 Z9 45 U1 0 U2 2 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 NOV PY 2000 VL 183 IS 5 BP 1070 EP 1077 DI 10.1067/mob.2000.108885 PG 8 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 377KD UT WOS:000165512000004 PM 11084543 ER PT J AU Davies, JK Shikes, RH Sze, CI Leslie, KK McDuffie, RS Romero, R Gibbs, RS AF Davies, JK Shikes, RH Sze, CI Leslie, KK McDuffie, RS Romero, R Gibbs, RS TI Histologic inflammation in the maternal and fetal compartments in a rabbit model of acute intra-amniotic infection SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article; Proceedings Paper CT 20th Annual Meeting of the Society-for-Maternal-Fetal-Medicine CY JAN 31-FEB 05, 2000 CL MIAMI BEACH, FLORIDA SP Soc Maternal Fetal Med DE intrauterine infection; histologic inflammation; rabbit; pregnancy ID NECROSIS-FACTOR-ALPHA; WHITE-MATTER LESIONS; BRONCHOPULMONARY DYSPLASIA; PREMATURE RUPTURE; CEREBRAL-PALSY; CYTOKINES INTERLEUKIN-6; PRETERM INFANTS; FLUID; CHORIOAMNIONITIS; MEMBRANES AB OBJECTIVE: This study was undertaken to determine the course of acute inflammation in the maternal and fetal compartments during experimentally induced ascending intra-amniotic infection. STUDY DESIGN: Forty pregnant rabbits at 70% gestation were inoculated endocervically with 10(5) colony-forming units of Escherichia coli. Does were killed at 0, 4, 8, 16, 24, and 30 hours after inoculation. At necropsy, blood, peritoneal fluid, amniotic fluid, and uterine tissue were cultured. Fetal brain, lung, heart, gut, and kidney were collected for histologic examination. Necrosis, infiltrates, congestion, and edema were each assessed semiquantitatively, and mean composite histologic-inflammation scores were compared with analysis of variance. Inflammation. mitotic activity, and apoptosis were evaluated in the fetal brain, and groups were compared with analysis of variance. RESULTS: Twenty-six animals were evaluated after 14 were excluded (lack of fever or positive culture results). A significant increase in histologic inflammation score was seen in the uterus (P < .001), placenta (P = .011), and fetal lung (P = .001) but not in other fetal tissues. These changes were seen earlier in the uterus and placenta and later in the fetal lung. Mitotic activity in the fetal brain decreased significantly by 8 hours after cervical inoculation. There was no inflammation in the fetal brain, and apoptosis in the fetal brain did not increase with time. CONCLUSIONS: Histologic inflammation occurs early in both the uterus and the placenta and later in the fetal lung in the rabbit model of acute intra-amniotic infection. This contrasts with the previously reported chronic model of intra-amniotic infection in the rabbit. C1 Univ Colorado, Hlth Sci Ctr, Dept Obstet & Gynecol, Boulder, CO 80309 USA. Univ Colorado, Hlth Sci Ctr, Dept Pathol, Boulder, CO 80309 USA. NICHHD, Perinatol Res Branch, Bethesda, MD 20892 USA. RP Davies, JK (reprint author), Univ Colorado, Hlth Sci Ctr, Dept Obstet & Gynecol, Boulder, CO 80309 USA. FU NIMHD NIH HHS [263-MD-731641-1] NR 25 TC 35 Z9 35 U1 0 U2 2 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 NOV PY 2000 VL 183 IS 5 BP 1088 EP 1093 DI 10.1067/mob.2000.108888 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 377KD UT WOS:000165512000007 PM 11084546 ER PT J AU Pacora, P Romero, R Maymon, E Gervasi, MT Gomez, R Edwin, SS Yoon, BH AF Pacora, P Romero, R Maymon, E Gervasi, MT Gomez, R Edwin, SS Yoon, BH TI Participation of the novel cytokine interleukin 18 in the host response to intra-amniotic infection SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article; Proceedings Paper CT 20th Annual Meeting of the Society-for-Maternal-Fetal-Medicine CY JAN 31-FEB 05, 2000 CL MIAMI BEACH, FLORIDA SP Soc Maternal Fetal Med DE interleukin 18; microbial invasion of the amniotic cavity; parturition; premature rupture of membranes; preterm labor ID GAMMA-INDUCING FACTOR; INDUCED LIVER-INJURY; IL-18; MICE; CELLS AB OBJECTIVE: Interleukin 18 is a proinflammatory pleiotropic cytokine that has been implicated in the host defense against infection. This study was undertaken to determine whether interleukin 18 concentrations change in the maternal, fetal, and amniotic fluid compartments with labor (term and preterm) and microbial invasion of the amniotic cavity. STUDY DESIGN: Amniotic fluid was assayed for interleukin 18 in samples obtained from 285 patients in the following groups: (1) term not in labor (n = 22), in labor (n = 19), and with microbial invasion of the amniotic cavity (n = 16); (2) preterm labor who delivered at term (n = 38), who delivered preterm but without microbial invasion of the amniotic cavity (n = 41), and preterm labor with microbial invasion of the amniotic cavity (n = 24); (3) preterm premature rupture of membranes without microbial invasion of the amniotic cavity (n = 30) and with microbial invasion of the amniotic cavity (n = 34); (4) term premature rupture of membranes not in labor (n = 20) and term premature rupture of membranes in labor (n = 19); and (5) midtrimester (n = 22). In addition, cord and maternal plasma samples from women at term not in labor (n = 20) and in labor (n = 20) were assayed for interleukin 18. RESULTS: (1) Interleukin 18 was detectable in all amniotic fluid samples and maternal and umbilical cord blood samples. (2) Interleukin 18 concentrations increased with advancing gestational age (r = 0.47; P < .0001). (3) Microbial invasion of the amniotic cavity in either preterm or term parturition was associated with a significant increase in the amniotic fluid concentration of interleukin 18 (preterm labor without microbial invasion of the amniotic cavity: median, 14.95 pg/mL; range, 3.9-277.0 pg/mL; vs preterm labor with microbial invasion of the amniotic cavity: median, 20.75 pg/mL; range, 5.53-160.21 pg/mL; P < .02; term labor without microbial invasion of the amniotic cavity: median, 18.73 pg/mL; range, 5.09-95.44 pg/mL; vs term labor with microbial invasion of the amniotic cavity: median, 24.35 pg/mL; range, 10.07-144.42 pg/mL; P < .004). (4) Both term and preterm parturition were associated with a modest increase in amniotic fluid interleukin 18 concentrations, although this trend did not reach statistical significance. (5) Rupture of membranes at term was associated with a significant decrease in amniotic fluid interleukin 18 concentrations (intact membranes: median, 14.96 pg/mL; range, <3.89-26.07 pg/mL; vs rupture of membranes: median, 10.1 pg/mL; range, 4.29-21.44 pg/mL; P < .001). CONCLUSION: (1) Interleukin 18 is increased in cases of microbial invasion of the amniotic cavity. (2) Interleukin 18 is detectable in the amniotic, maternal, and fetal compartments. (3) We propose that this novel cytokine plays a role in the host defense against infection. C1 Wayne State Univ, Hutzel Hosp, Dept Obstet & Gynecol, NICHHD,Perinatol Res Branch, Detroit, MI 48201 USA. Seoul Natl Univ, Dept Obstet & Gynecol, Seoul 151, South Korea. RP Romero, R (reprint author), Wayne State Univ, Hutzel Hosp, Dept Obstet & Gynecol, NICHHD,Perinatol Res Branch, 4707 St Antoine Blvd, Detroit, MI 48201 USA. RI Yoon, Bo Hyun/H-6344-2011 NR 12 TC 61 Z9 62 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 NOV PY 2000 VL 183 IS 5 BP 1138 EP 1143 DI 10.1067/mob.2000.108881 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 377KD UT WOS:000165512000016 PM 11084555 ER PT J AU Hartge, P Hayes, R Reding, D Sherman, ME Prorok, P Schiffman, M Buys, S AF Hartge, P Hayes, R Reding, D Sherman, ME Prorok, P Schiffman, M Buys, S TI Complex ovarian cysts in postmenopausal women are not associated with ovarian cancer risk factors - Preliminary data from the Prostate, Lung, Colon, and Ovarian Cancer Screening Trial SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE ovarian cancer; ovarian cysts; epidemiology ultrasonograph; screening; precursors AB OBJECTIVE: We assessed whether asymptomatic ovarian abnormalities detected on ultrasonography in postmenopausal women are precursors to ovarian cancer. STUDY DESIGN: We compared the transvaginal ultrasonographic findings from the initial examination of 20,000 postmenopausal women enrolled to date in an ongoing randomized trial of cancer screening with data on the established risk factors for ovarian cancer obtained from self-administered questionnaires. We distinguished cysts with the suggestive characteristic(s) of a septum, a solid component, or an irregular or thick wall ("complex cysts") from simple sonolucent cysts with none of those features. RESULTS: High parity, a strong ovarian cancer protective factor, was negatively associated with complex cysts (odds ratio for greater than or equal to5 births vs no births, 0.72; 95% confidence interval, 0.53-0.97), but long-term oral contraceptive use, another strong ovarian cancer protective factor, was not associated with complex cysts (odds ratio, 0.96; 95% confidence interval, 0.76-1.20). A family history of ovarian cancer or multiple breast cancers, a strong risk factor for cancer, was not associated with complex cysts (odds ratio, 0.99; 95% confidence interval, 0.68-1.44). Other abnormalities found on ultrasonography (including simple cysts, bilateral cysts, or all abnormalities combined) also did not share the established risk factors for ovarian malignancy. We did not identify any combination of features of abnormalities (septum, echogenicity, size, or papillary projections) that manifested the cancer risk factor profile. CONCLUSIONS: Although a very small proportion of the clinically silent ovarian abnormalities found on ultrasonography are determined to be ovarian cancers, the remaining complex cysts and other clinically suspicious abnormalities do not appear to be the immediate precursors of ovarian cancer. The eventual identification of such precursors will yield opportunities for earlier diagnosis, screening of high-risk groups, and better understanding of the cause of this often lethal malignancy. C1 NCI, Epidemiol & Biostat Program, Bethesda, MD 20892 USA. Univ Utah, Div Oncol, Salt Lake City, UT 84112 USA. Marshfield Med Res Fdn, Marshfield, WI USA. RP Hartge, P (reprint author), NCI, Epidemiol & Biostat Program, 6120 Execut Blvd,EPS 8090, Bethesda, MD 20892 USA. NR 11 TC 25 Z9 26 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 NOV PY 2000 VL 183 IS 5 BP 1232 EP 1237 DI 10.1067/mob.2000.107401 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 377KD UT WOS:000165512000032 PM 11084571 ER PT J AU Mohr, VH Vortmeyer, AO Zhuang, ZP Libutti, SK Walther, MM Choyke, PL Zbar, B Linehan, WM Lubensky, IA AF Mohr, VH Vortmeyer, AO Zhuang, ZP Libutti, SK Walther, MM Choyke, PL Zbar, B Linehan, WM Lubensky, IA TI Histopathology and molecular genetics of multiple cysts and microcystic (serous) adenomas of the pancreas in von Hippel-Lindau patients SO AMERICAN JOURNAL OF PATHOLOGY LA English DT Article ID TUMOR-SUPPRESSOR GENE; RENAL-CELL CARCINOMA; NEUROENDOCRINE TUMORS; DISEASE PATIENTS; LESIONS; CYSTADENOMA; DELETION; PREVALENCE; FEATURES; MEMBERS AB Microcystic adenoma and cysts of the pancreas occur sporadically or as a part of von Hippel-Lindau (VHL) disease, The pathology of pancreatic cystic disease in VHL patients has not been well characterized. Furthermore, it is presently unknown whether the alteration of the VHL gene is responsible for the development of the entire spectrum of pancreatic serous cystic lesions. We performed a histopathological analysis of 21 cysts and 98 microcystic adenomas in nine VHL patients with a known germline mutation. In addition, PCR-amplified DNA from 27 pancreatic cystic lesions in three informative patients was studied for allelic deletions with polymorphic markers spanning the VHL. gene locus. In all patients, pancreatic lesions were multiple: 21 benign serous cysts, 63 microscopic microcystic adenomas (size <0.4 cm), and 35 macroscopic microcystic adenomas (size >0.5 cm). The average number of lesions per patient was 2.1 benign cysts (range, 0-8), 7.7 (1-37) microscopic microcystic adenomas, and 3 (0-21) macroscopic microcystic adenomas. All lesions showed similar histology and contained prominent fibrous stroma, clear and/or amphophilic, glycogen-rich epithelial cells, endothelial and smooth muscle cells. VHL, deletions were detected In all types of pancreatic cystic lesions. The presence of VHL gene allelic deletions in the spectrum of multifocal pancreatic cystic lesions provides direct molecular evidence of their neoplastic nature and integral association with VHL disease. The histopathological and molecular data establish a serous cyst-microcystic adenoma continuum in the development of pancreatic cystic neoplasia in VHL disease. C1 NINDS, Mol Pathogenesis Unit, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, Bethesda, MD 20892 USA. NCI, Surg Branch, Bethesda, MD 20892 USA. NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. NIH, Warren G Magnuson Clin Ctr, Dept Radiol, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Immunobiol Lab, Frederick, MD 21701 USA. RP Lubensky, IA (reprint author), NINDS, Mol Pathogenesis Unit, Surg Neurol Branch, NIH, Bldg 10,Room 5D37,10 Ctr Dr, Bethesda, MD 20892 USA. NR 36 TC 74 Z9 78 U1 0 U2 3 PU AMER SOC INVESTIGATIVE PATHOLOGY, INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3993 USA SN 0002-9440 J9 AM J PATHOL JI Am. J. Pathol. PD NOV PY 2000 VL 157 IS 5 BP 1615 EP 1621 DI 10.1016/S0002-9440(10)64799-2 PG 7 WC Pathology SC Pathology GA 372GU UT WOS:000165226600021 PM 11073821 ER PT J AU Jayadev, S Barrett, JC Murphy, E AF Jayadev, S Barrett, JC Murphy, E TI Elevated ceramide is downstream of altered calcium homeostasis in low serum-induced apoptosis SO AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY LA English DT Article DE endoplasmic reticulum; thapsigargin; cell cycle arrest; diacylglycerol ID NECROSIS-FACTOR-ALPHA; NEOPLASTIC PROGRESSION; INTRACELLULAR CALCIUM; PROTEIN-KINASE; T-CELLS; SPHINGOMYELINASE; SPHINGOLIPIDS; IDENTIFICATION; STIMULATION; THYMOCYTES AB Two immortalized cell lines, sup (+) and sup (-), derived from mutagenized Syrian hamster embryo cells, were used to study the relationship and temporal order between calcium and ceramide signals during apoptosis. The early preneoplastic cells, termed sup (+), suppress tumorigenicity when hybridized with tumor cells, whereas later-stage sup (-) cells do not. In reduced serum conditions, sup (+) cells cease proliferating and undergo apoptosis; in contrast, sup (-) cells continue slow growth and undergo necrosis. In sup (+) cells, decreased endoplasmic reticulum (ER) calcium occurs 4 h after low serum treatment and precedes apoptosis. Significant elevations in ceramide are observed 16 h after reduced serum treatment of sup (+) cells but are not found in sup (-) cells. Inhibiting ER calcium depletion in low serum-treated sup (+) cells by treating with high levels of calcium prevents both ceramide generation and apoptosis. Conversely, inducing ER calcium depletion in sup (-) cells by treating with low serum plus thapsigargin results in elevated ceramide levels and apoptosis. Furthermore, C-6-ceramide treatment induced apoptosis of sup (-) cells in low serum, a condition that does not normally cause apoptosis. C-6-ceramide treatment did not induce apoptosis in either sup (+) or sup (-) cells in 10% serum but did cause G(2)/M arrest. These studies show that ceramide production is downstream of ER calcium release. C1 NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. RP Murphy, E (reprint author), NIEHS, Mol Carcinogenesis Lab, POB 12233,MD D2-03, Res Triangle Pk, NC 27709 USA. NR 44 TC 9 Z9 9 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6143 J9 AM J PHYSIOL-CELL PH JI Am. J. Physiol.-Cell Physiol. PD NOV PY 2000 VL 279 IS 5 BP C1640 EP C1647 PG 8 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 362QT UT WOS:000089790300037 PM 11029312 ER PT J AU Platoshyn, O Golovina, VA Bailey, CL Limsuwan, A Krick, S Juhaszova, M Seiden, JE Rubin, LJ Yuan, JXJ AF Platoshyn, O Golovina, VA Bailey, CL Limsuwan, A Krick, S Juhaszova, M Seiden, JE Rubin, LJ Yuan, JXJ TI Sustained membrane depolarization and pulmonary artery smooth muscle cell proliferation SO AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY LA English DT Article DE intracellular calcium; voltage-gated potassium channels; membrane potential ID GATED K+ CHANNELS; DEPENDENT PROTEIN-KINASE; HUMAN LYMPHOCYTE-T; GENE-EXPRESSION; POTASSIUM CHANNELS; NITRIC-OXIDE; CA2+ RELEASE; CALCIUM; HYPERTENSION; ACTIVATION AB Pulmonary vasoconstriction and vascular medial hypertrophy greatly contribute to the elevated pulmonary vascular resistance in patients with pulmonary hypertension. A rise in cytosolic free Ca2+ ([ Ca2+](cyt)) in pulmonary artery smooth muscle cells (PASMC) triggers vasoconstriction and stimulates cell growth. Membrane potential (E-m) regulates [Ca2+](cyt) by governing Ca2+ influx through voltage-dependent Ca2+ channels. Thus intracellular Ca2+ may serve as a shared signal transduction element that leads to pulmonary vasoconstriction and vascular remodeling. In PASMC, activity of voltage-gated K+ (Kv) channels regulates resting E-m. In this study, we investigated whether changes of Kv currents [I-K(V)], E-m, and [Ca2+](cyt) affect cell growth by comparing these parameters in proliferating and growth-arrested PASMC. Serum deprivation induced growth arrest of PASMC, whereas chelation of extracellular Ca2+ abolished PASMC growth. Resting [Ca2+](cyt) was significantly higher, and resting E-m was more depolarized, in proliferating PASMC than in growth-arrested cells. Consistently, whole cell I-K(V) was significantly attenuated in PASMC during proliferation. Furthermore, E-m depolarization significantly increased resting [Ca2+](cyt) and augmented agonist-mediated rises in [Ca2+](cyt) in the absence of extracellular Ca2+. These results demonstrate that reduced I-K(V), depolarized E-m, and elevated [Ca2+](cyt) may play a critical role in stimulating PASMC proliferation. Pulmonary vascular medial hypertrophy in patients with pulmonary hypertension may be partly caused by a membrane depolarization-mediated increase in [Ca2+](cyt) in PASMC. C1 Univ Calif San Diego, Med Ctr, Dept Med, Sch Med, San Diego, CA 92103 USA. Univ Maryland, Dept Physiol, Baltimore, MD 21201 USA. Univ Maryland, Dept Med, Baltimore, MD 21201 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Yuan, JXJ (reprint author), Univ Calif San Diego, Med Ctr, Dept Med, Sch Med, MC 8382,200 W Arbor Dr, San Diego, CA 92103 USA. FU NHLBI NIH HHS [HL54043, HL64945] NR 66 TC 104 Z9 118 U1 0 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6143 J9 AM J PHYSIOL-CELL PH JI Am. J. Physiol.-Cell Physiol. PD NOV PY 2000 VL 279 IS 5 BP C1540 EP C1549 PG 10 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 362QT UT WOS:000089790300026 PM 11029301 ER PT J AU Sweet, DH Miller, DS Pritchard, JB AF Sweet, DH Miller, DS Pritchard, JB TI Basolateral localization of organic cation transporter 2 in intact renal proximal tubules SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE green fluorescent protein; kidney; killifish; transfection; Madin-Darby canine kidney cells; LLC-PK(1) cells ID FUNCTIONAL EXPRESSION; DEPENDENT TRANSPORT; ANION; SECRETION; MEMBRANE; OCT2; MECHANISMS; CLONING AB The localization of organic cation transporter 2 (OCT2) within renal cells is the subject of considerable controversy, resulting in marked uncertainty as to its function. To resolve this issue, we made an OCT2/green fluorescent protein (GFP) fusion construct (rOCT2-GFP) and determined its localization within Xenopus laevis oocytes and renal cells using confocal microscopy. Oocytes expressing rOCT2-GFP exhibited plasma membrane fluorescence as well as greatly increased specific, potential-driven uptake of [(14)C] tetraethylammonium (TEA). Polarized monolayers of renal epithelial cell lines [LLC-PK(1) and Madin-Darby canine kidney (MDCK)] transiently transfected with pEGFP-C3, which codes for a cytoplasmic GFP, showed a diffuse, evenly distributed cytoplasmic signal with no plasma membrane fluorescence. In contrast, cells transiently transfected with pEGFP-C3/rOCT2 (the vector coding for rOCT2-GFP) showed predominantly plasma membrane fluorescence, which was most prominent in the lateral membrane. MDCK cells stably expressing rOCT2-GFP (MDCK/rOCT2-GFP) maintained in long-term culture showed a greatly increased basal and lateral membrane fluorescence. When grown on porous supports, MDCK/rOCT2-GFP monolayers showed specific, potential-driven TEA uptake from the basal side. Finally, expression and distribution of rOCT2-GFP were investigated in isolated killifish (Fundulus heteroclitus) renal proximal tubules. On expression of rOCT2-GFP, transfected tubules showed marked basal and lateral membrane fluorescence, with no detectable signal at the apical membrane. In contrast, tubules expressing a luminal sodium-dicarboxylate cotransporter (rbNaDC-1)-GFP construct showed apical membrane fluorescence, and tubules expressing cytoplasmic GFP had a diffuse cytoplasmic fluorescence. These results indicate that rOCT2 is basolateral in renal proximal tubule cells. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. RP Pritchard, JB (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, Mail Drop F1-03,POB 12233, Res Triangle Pk, NC 27709 USA. EM pritchard@niehs.nih.gov RI Sweet, Douglas/H-7914-2013 OI Sweet, Douglas/0000-0002-8911-9184 NR 22 TC 42 Z9 42 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 NOV PY 2000 VL 279 IS 5 BP F826 EP F834 PG 9 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 367LM UT WOS:000090063800005 PM 11053042 ER PT J AU Yang, TX Endo, Y Huang, YG Smart, A Briggs, JP Schnermann, J AF Yang, TX Endo, Y Huang, YG Smart, A Briggs, JP Schnermann, J TI Renin expression in COX-2-knockout mice on normal or low-salt diets SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE renin activity; prostaglandins; macula densa; juxtaglomerular apparatus cyclooxygenase-2 ID MACULA DENSA; RAT-KIDNEY; CYCLOOXYGENASE-2 EXPRESSION; RENOVASCULAR HYPERTENSION; DIFFERENTIAL REGULATION; PROSTAGLANDIN SYNTHESIS; NUCLEOTIDE-SEQUENCE; GENE DISRUPTION; MESSENGER-RNA; CYCLIC-AMP AB Experiments were performed in mice to investigate whether cyclooxygenase-2 (COX-2) in epithelial cells near the tubulovascular contact point (macula densa and TAL cells) may regulate renin gene expression in juxtaglomerular granular cells. Renin activity, afferent arteriolar granularity, and renin mRNA were determined in wild-type mice and in COX-2-knockout mice on control and low-NaCl diets. Renin activity in microdissected glomeruli assessed as angiotensin I formation in the presence of excess substrate and afferent arteriolar granularity determined by direct visualization and immunostaining were significantly reduced in COX-2 -/- compared with wild-type animals. Similarly, renal cortical mRNA levels were lower in COX-2 -/- than in wild-type mice. Maintaining mice on a low-salt diet for 14 days induced an increase in renin mRNA, afferent arteriolar granularity, and renin activity in wild-type mice. In contrast, renin mRNA and renin granularity did not significantly increase in low-salt-treated COX-2 -/- mice, whereas the increase in juxtaglomerular renin enzyme activity was markedly attenuated, but not fully blocked. In additional experiments we found that COX-2 mRNA was increased in angiotensin type 1A receptor-knockout mice compared with wild-type mice. We conclude that COX-2 in the tubulovascular contact region is a critical determinant of renin synthesis in granular cells under resting conditions and that it participates in the stimulation of renin expression caused by a low-NaCl intake. C1 NIDDKD, NIH, Bethesda, MD 20892 USA. RP Schnermann, J (reprint author), NIDDKD, NIH, Bldg 10,Rm 4 D51,10 Ctr Dr MSC 1370, Bethesda, MD 20892 USA. EM jurgens@intra.niddk.nih.gov RI Briggs, Josephine/B-9394-2009 OI Briggs, Josephine/0000-0003-0798-1190 NR 41 TC 76 Z9 76 U1 0 U2 2 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 NOV PY 2000 VL 279 IS 5 BP F819 EP F825 PG 7 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 367LM UT WOS:000090063800004 PM 11053041 ER PT J AU Hyman, SE AF Hyman, SE TI The needs for database research and for privacy collide SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Editorial Material C1 NIMH, Bethesda, MD 20892 USA. RP Hyman, SE (reprint author), NIMH, 31 Ctr Dr,Room 4A-5, Bethesda, MD 20892 USA. NR 0 TC 4 Z9 4 U1 0 U2 1 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 NOV PY 2000 VL 157 IS 11 BP 1723 EP 1724 DI 10.1176/appi.ajp.157.11.1723 PG 2 WC Psychiatry SC Psychiatry GA 369YJ UT WOS:000165095100001 PM 11058462 ER PT J AU Pietrini, P Guazzelli, M Basso, G Jaffe, K Grafman, J AF Pietrini, P Guazzelli, M Basso, G Jaffe, K Grafman, J TI Neural correlates of imaginal aggressive behavior assessed by positron emission tomography in healthy subjects SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article; Proceedings Paper CT European Regional Congress of the World-Federation-Societies-of-Biological-Psychiatry CY APR 08-11, 1999 CL FLORENCE, ITALY SP World Federat Soc ID CEREBRAL BLOOD-FLOW; SCRIPT-DRIVEN IMAGERY; POSTTRAUMATIC-STRESS-DISORDER; OBSESSIVE-COMPULSIVE DISORDER; HUMAN PREFRONTAL CORTEX; FRONTAL-LOBE; SYMPTOM PROVOCATION; ALZHEIMERS-DISEASE; GLUCOSE-METABOLISM; EMOTIONAL VALENCE AB objective: Neurodegenerative or traumatic lesions of the frontal lobes often lead to abnormally aggressive behavior. The authors hypothesized that the imaginal evoking of scenarios involving aggressive behavior would be associated with a modulation of the functional activity in the human frontal cortex. Method: Regional cerebral blood flow (rCBF) determinations by positron emission tomography and psychophysiological measures of emotional responsivity were obtained in a group of 15 young healthy Volunteers with good visual imagery abilities and no history of abnormal behavior while they imagined the same scenario with four variations involving emotionally neutral behavior and aggressive behavior. Results: Compared to the imagined neutral scenario, the imagined scenarios involving aggressive behavior were associated with significant emotional reactivity and rCBF reductions in the ventromedial prefrontal cortex, suggesting that a functional deactivation of this cortical area occurs when individuals respond to the eliciting of imagined aggressive behavior. Conclusions: These results in healthy subjects further expand previous findings from animal and human studies by providing an in vivo functional demonstration of the involvement of the orbitofrontal cortex in the expression of aggressive behavior. They are also consistent with the hypothesis that a functional alteration of this cortical region may be present in individuals with pathological aggressive behavior. C1 NINDS, Cognit Neurosci Sect, NIH, Bethesda, MD 20892 USA. Univ Pisa, Dept Human & Environm Sci, I-56100 Pisa, Italy. Univ Pisa, Dept Psychiat, I-56100 Pisa, Italy. Univ Pisa, Dept Pharmacol, I-56100 Pisa, Italy. Univ Pisa, Dept Neurobiol & Biotechnol, I-56100 Pisa, Italy. RP Pietrini, P (reprint author), NINDS, Cognit Neurosci Sect, NIH, Bldg 10,Rm 5C205,10 Ctr Dr,MSC 1440, Bethesda, MD 20892 USA. RI Basso, Gianpaolo/A-9208-2012; OI Basso, Gianpaolo/0000-0002-6245-9402; Grafman, Jordan H./0000-0001-8645-4457 NR 83 TC 138 Z9 143 U1 11 U2 13 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 NOV PY 2000 VL 157 IS 11 BP 1772 EP 1781 DI 10.1176/appi.ajp.157.11.1772 PG 10 WC Psychiatry SC Psychiatry GA 369YJ UT WOS:000165095100012 PM 11058474 ER PT J AU Hirschfeld, RMA Williams, JBW Spitzer, RL Calabrese, JR Flynn, L Keck, PE Lewis, L McElroy, SL Post, RM Rapport, DJ Russell, JM Sachs, GS Zajecka, J AF Hirschfeld, RMA Williams, JBW Spitzer, RL Calabrese, JR Flynn, L Keck, PE Lewis, L McElroy, SL Post, RM Rapport, DJ Russell, JM Sachs, GS Zajecka, J TI Development and validation of a screening instrument for bipolar spectrum disorder: The mood disorder questionnaire SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article AB Objective: Bipolar spectrum disorders, which include bipolar I, bipolar II, and bipolar disorder not otherwise specified, frequently go unrecognized, undiagnosed, and untreated. This report describes the validation of a new brief self-report screening instrument for bipolar spectrum disorders called the Mood Disorder Questionnaire. Method: A total of 198 patients attending five outpatient clinics that primarily treat patients with mood disorders completed the Mood Disorder Questionnaire. A research professional, blind to the Mood Disorder Questionnaire results, conducted a telephone research diagnostic interview by means of the bipolar module of the Structured Clinical Interview for DSM-IV. Results: A Mood Disorder Questionnaire screening score of 7 or more items yielded good sensitivity (0.73) and very good specificity (0.90). Conclusions: The Mood Disorder Questionnaire is a useful screening instrument for bipolar spectrum disorder in a psychiatric outpatient population. C1 Univ Texas, Dept Psychiat & Behav Sci, Med Branch, Galveston, TX 77555 USA. Columbia Univ, Dept Psychiat, New York, NY USA. New York State Psychiat Inst, New York, NY 10032 USA. Case Western Reserve Univ, Sch Med, Cleveland, OH USA. Natl Assoc Mentally Ill, Washington, DC USA. Univ Cincinnati, Coll Med, Dept Psychiat, Biol Psychiat Program, Cincinnati, OH USA. Natl Depress & Man Depress Assoc, Chicago, IL USA. NIMH, Bethesda, MD 20892 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Boston, MA USA. Rush Presbyterian St Lukes Med Ctr, Chicago, IL 60612 USA. RP Hirschfeld, RMA (reprint author), Univ Texas, Dept Psychiat & Behav Sci, Med Branch, 1-302 Rebecca Sealy,301 Univ Blvd, Galveston, TX 77555 USA. NR 8 TC 664 Z9 679 U1 2 U2 36 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 NOV PY 2000 VL 157 IS 11 BP 1873 EP 1875 DI 10.1176/appi.ajp.157.11.1873 PG 3 WC Psychiatry SC Psychiatry GA 369YJ UT WOS:000165095100028 PM 11058490 ER PT J AU Helfand, WH Lazarus, J Theerman, P AF Helfand, WH Lazarus, J Theerman, P TI The Children's Bureau and public health at midcentury SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Article C1 Natl Lib Med, Hist Med Div, Bethesda, MD 20894 USA. RP Theerman, P (reprint author), Natl Lib Med, Hist Med Div, 8600 Rockville Pike, Bethesda, MD 20894 USA. NR 1 TC 0 Z9 0 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 NOV PY 2000 VL 90 IS 11 BP 1703 EP 1703 DI 10.2105/AJPH.90.11.1703 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 369FJ UT WOS:000165055000005 PM 11076234 ER PT J AU Koss, MP Goldman, D AF Koss, MP Goldman, D TI Genetic factors and alcoholism SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Letter C1 Univ Arizona, Coll Publ Hlth, Div Hlth Promot Sci, Tucson, AZ USA. NIAAA, Neurogenet Lab, Rockville, MD 20852 USA. RP Koss, MP (reprint author), Ten Tribes Study, 1632 E Lester St, Tucson, AZ 85719 USA. NR 1 TC 1 Z9 1 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 NOV PY 2000 VL 90 IS 11 BP 1799 EP 1799 DI 10.2105/AJPH.90.11.1799 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 369FJ UT WOS:000165055000032 PM 11076256 ER PT J AU Berhane, K McConnell, R Gilliland, F Islam, T Gauderman, WJ Avol, E London, SJ Rappaport, E Margolis, HG Peters, JM AF Berhane, K McConnell, R Gilliland, F Islam, T Gauderman, WJ Avol, E London, SJ Rappaport, E Margolis, HG Peters, JM TI Sex-specific effects of asthma on pulmonary function in children SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article ID SOUTHERN CALIFORNIA COMMUNITIES; ADULT LUNG-FUNCTION; VENTILATORY FUNCTION; CHILDHOOD ASTHMA; WHEEZING ILLNESS; DIFFERING LEVELS; AIR-POLLUTION; FOLLOW-UP; GROWTH; POPULATION AB To evaluate the effects on lung function of asthma, time since diagnosis of asthma, and age at diagnosis of asthma, we examined school children in a cohort of 2,277 fourth- and seventh-graders at least twice during a 4-yr follow-up period. Sex-specific models for each lung function were fitted through mixed-effects models that used regression splines and captured age-dependent trends in the effect of asthma on lung function. In males, a history of asthma was associated with large and statistically significant deficits in maximum midexpiratory flow (MMEF) (-4.89%) and forced expiratory flow at 75% of expired FVC (FEF75) (-6.62%), whereas in females these deficits were smaller (-1.93% and -2.45%, respectively) and were not statistically significant. However, larger deficits were seen in both males and females with longer time since diagnosis. In males with more than 6 yr since diagnosis, there were significant deficits in FEV1 (-3.91%), MMEF (-7.39%), FEF75 (-8.12%), and peak expiratory flow rate (PEFR) (-4.65%) as compared with children with less than 3 yr since diagnosis. There were fewer females with more than 6 yr since diagnosis, but deficits were similar to those of males for FEV1 (-2.52%), MMEF (-9.26%), and FEF75 (-14.28%). Large deficits in flow rates in both large and small airways were observed in males and females for whom asthma was reported to have been diagnosed before age 3 yr. There was little evidence that lung growth in children with asthma "catches up" at older ages. Therefore, because a constant percent deficit in lung function implies an increasingly large absolute deficit in older children with larger lungs, these results are consistent with prior evidence that lung function deficits in children with asthma persist into adulthood. We also suggest that in children, commonly observed differences between sexes in the impact of asthma on lung function may reflect differences in the duration and age of onset of asthma in males and females. C1 Univ So Calif, Sch Med, Dept Prevent Med, Los Angeles, CA 90033 USA. NIEHS, Res Triangle Pk, NC 27709 USA. Calif Air Resources Board, Sacramento, CA USA. RP Berhane, K (reprint author), Univ So Calif, Keck Sch Med, Dept Prevent Med, 1540 Alcazar St,CHP 220, Los Angeles, CA 90033 USA. OI London, Stephanie/0000-0003-4911-5290 FU NHLBI NIH HHS [1R01HL61768-01]; NIEHS NIH HHS [1P01ES0939581-01, 5P30ES07048-04] NR 32 TC 34 Z9 35 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 NOV PY 2000 VL 162 IS 5 BP 1723 EP 1730 PG 8 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 371WU UT WOS:000165202900023 PM 11069803 ER PT J AU Crapo, JD Harmsen, AG Sherman, MP Musson, RA AF Crapo, JD Harmsen, AG Sherman, MP Musson, RA TI Pulmonary immunobiology and inflammation in pulmonary diseases SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article ID HUMAN-LUNG PARENCHYMA; DENDRITIC CELLS; PHOSPHOLIPASE A(2); RESPIRATORY-TRACT; HOST-DEFENSE; T-CELLS; ANTIGEN; IMMUNE; COMPARTMENTALIZATION; LYMPHOCYTES C1 NHLBI, Rockledge Ctr 2, Div Lung Dis, Bethesda, MD 20892 USA. RP Musson, RA (reprint author), NHLBI, Rockledge Ctr 2, Div Lung Dis, Suite 10018,6701 Rockledge Dr,MSC 7952, Bethesda, MD 20892 USA. NR 35 TC 34 Z9 36 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 NOV PY 2000 VL 162 IS 5 BP 1983 EP 1986 PG 4 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 371WU UT WOS:000165202900064 PM 11069844 ER PT J AU Man, YG Martinez, A Avis, IM Hong, SH Cuttitta, F Venzon, DJ Mulshine, JL AF Man, YG Martinez, A Avis, IM Hong, SH Cuttitta, F Venzon, DJ Mulshine, JL TI Phenotypically different cells with heterogeneous nuclear ribonucleoprotein A2/B1 overexpression show similar genetic alterations SO AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY LA English DT Article ID X-CHROMOSOME INACTIVATION; LUNG-CANCER DETECTION; BRONCHIAL EPITHELIUM; FORMER SMOKERS; EXPRESSION; HNRNP; PROTEIN; ANTIGEN; REPEAT; TUMORS AB Immunocytochemical studies have revealed that overexpression of heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1 in exfoliated epithelial cells is a potentially useful marker of early lung cancer. This study analyzed the correlation of hnRNP A2/B1 expression with molecular alterations in phenotypically different epithelial cells of paraffin-embedded pulmonary tissues. Sections from 20 human subjects were analyzed immunohistochemically for expression of hnRNP A2/B1, Normal-appearing, hyperplastic, and malignant epithelial cells with and without hnRNP A2/B1 expression (n = 78) were microdissected and assessed for microsatellite alterations (MA) and loss of heterozygosity (LOH) (n = 14 markers) as well as for clonality. Results showed that (I) hnRNP A2/B1 immunoreactive cells contained a significantly higher frequency of MA and LOH than did comparable cells that lacked detectable hnRNP A2/B1; (2) over 80% of MA and LOH seen in hnRNP A2/B1 immunoreactive normal-appearing and hyperplastic cells persisted in malignant cells; (3) preliminary analysis of methylation status of the androgen receptor gene in non-neoplastic cells was suggestive of hnRNP A2/B1-expressing cells being of clonal origin; and (4) cells with cytoplasmic hnRNP A2/B1 immunoreactivity had a 3-fold higher frequency of MA and LOH than did cells with nuclear hnRNP A2/B1 immunoreactivity. These findings suggest that phenotypically different respiratory epithelial cells with hnRNP A2/B1 overexpression might be clonally derived, and that the subcellular localization of hnRNP A2/B1 might be an important factor associated with tumor progression. C1 NCI, Div Clin Sci, Med Branch,Intervent Sect, Dept Cell & Canc Biol, Bethesda, MD 20892 USA. NCI, Div Clin Sci, Med Branch, Biostat & Data Management Sect, Bethesda, MD 20892 USA. RP Mulshine, JL (reprint author), NCI, Div Clin Sci, Med Branch,Intervent Sect, Dept Cell & Canc Biol, Bldg 10, Bethesda, MD 20892 USA. RI Venzon, David/B-3078-2008; Martinez, Alfredo/A-3077-2013 OI Martinez, Alfredo/0000-0003-4882-4044 NR 33 TC 28 Z9 36 U1 0 U2 3 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1044-1549 J9 AM J RESP CELL MOL JI Am. J. Respir. Cell Mol. Biol. PD NOV PY 2000 VL 23 IS 5 BP 636 EP 645 PG 10 WC Biochemistry & Molecular Biology; Cell Biology; Respiratory System SC Biochemistry & Molecular Biology; Cell Biology; Respiratory System GA 373FA UT WOS:000165276700008 PM 11062142 ER PT J AU Hamadeh, H Afshari, CA AF Hamadeh, H Afshari, CA TI Gene chips and functional genomics - A new technology will allow environmental health scientists to track the expression of thousands of genes in a single, fast and easy test SO AMERICAN SCIENTIST LA English DT Article ID CDNA MICROARRAYS; DISCOVERY C1 NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. RP Hamadeh, H (reprint author), NIEHS, Mol Carcinogenesis Lab, POB 12233, Res Triangle Pk, NC 27709 USA. NR 17 TC 14 Z9 15 U1 0 U2 1 PU SIGMA XI-SCI RES SOC PI RES TRIANGLE PK PA PO BOX 13975, RES TRIANGLE PK, NC 27709 USA SN 0003-0996 J9 AM SCI JI Am. Scientist PD NOV-DEC PY 2000 VL 88 IS 6 BP 508 EP 515 DI 10.1511/2000.41.874 PG 8 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 364GH UT WOS:000089883700016 ER PT J AU Territo, PR Balaban, RS AF Territo, PR Balaban, RS TI Rapid spectrophotometric determination of oxygen consumption using hemoglobin, in vitro: Light scatter correction and expanded dynamic range SO ANALYTICAL BIOCHEMISTRY LA English DT Article DE metabolism; spectrophotometric; light scattering; hemoglobin; response time ID PHOTOLYSIS AB The method of using absorbance in conjunction with hemoglobin (Hb) to monitor rapid changes in oxygen consumption in vitro was improved by using a nonlinear calibration technique and multiwavelength spectroscopy. The O-2 dependence of Hb absorbance was effectively linearized using the current technique (R-2 = 0.990 +/- 0.0021 n = 3), and extended the dynamic range of [O-2] determinations by 1.6-fold over previous approaches. The association/dissociation rates of O-2 and Hb were evaluated using the current approach and were not significant on the 100-ms time domain. A method was also developed for compensating for large amplitude light scattering changes in turbid media using multiwavelength analysis. Both the nonlinear calibration curve and light scattering corrections were validated in isolated porcine heart mitochondrial preparations. (C) 2000 Academic Press. C1 NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Territo, PR (reprint author), NHLBI, Cardiac Energet Lab, NIH, Bldg 10,Room B1D-416, Bethesda, MD 20892 USA. RI Balaban, Robert/A-7459-2009 OI Balaban, Robert/0000-0003-4086-0948 NR 13 TC 10 Z9 10 U1 0 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-2697 J9 ANAL BIOCHEM JI Anal. Biochem. PD NOV 1 PY 2000 VL 286 IS 1 BP 156 EP 163 DI 10.1006/abio.2000.4774 PG 8 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 371WA UT WOS:000165201200020 PM 11038286 ER PT J AU Tian, HJ Brody, LC Mao, D Landers, JP AF Tian, HJ Brody, LC Mao, D Landers, JP TI Effective capillary electrophoresis-based heteroduplex analysis through optimization of surface coating and polymer networks SO ANALYTICAL CHEMISTRY LA English DT Article ID DOUBLE-STRANDED DNA; ARRAY ELECTROPHORESIS; UNCOATED CAPILLARIES; MUTATION ANALYSIS; CONFORMATION POLYMORPHISM; FLUORESCENCE DETECTION; BRCA1; SEPARATION; TECHNOLOGY; DIAGNOSIS AB The efficacy of capillary electrophoresis for detecting DNA mutations via heteroduplex analysis (HDA) is dependent upon both the effective passivition of the capillary surface and the choice of the correct polymer network for sieving. Using HDA with laser-induced fluorescence detection of fluorescently labeled DNA fragments, an effective coating and optimal polymer matrix were sought. Optimized separation conditions were determined through the methodological evaluation of a number of different silanizing reagents, polymeric coatings, and polymer networks for resolving the PCR-amplified DNA fragments associated with five mutations (185delAG, 1294de140, 4446C>G, 5382insC, 5677insA) in the breast cancer susceptibility gene (BRCA1), For capillary coating, allyldimethylchlorosilane, 4-chlorobutyldimethylchlorosilane, (gamma -methacryloxypropyl)trinlethoxysilane, chlorodimethyloctylsilane (OCT), and 7-octenyltrimethoxysilane were evaluated as silanizing reagents in combination with poly(vinylpyrrolidone) (PVP) and polyacrylamide (PA) as the polymeric coat. The HDA results were compared with those obtained using a commercial (FC) coated capillary. Of these, the OCT-PVP combination was found to be most effective. Using this modified capillary, HDA with polymer networks that included hydroxyethylcellulose (HEC), linear polyacrylamide, and PVP showed that a PVP-, PA-, or FC-coated capillary, in combination with HEC as the sieving polymer, could be used effectively to discriminate the mutations in less than 10 min. However, optimal performance was observed with the OCT-PVP-coated capillary and HEC as the polymer network. C1 Univ Virginia, Dept Chem, Charlottesville, VA 22901 USA. Univ Pittsburgh, Dept Chem, Pittsburgh, PA 15260 USA. NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. J&W Sci, Folsom, CA 95630 USA. Univ Virginia, Med Ctr, Dept Pathol, Charlottesville, VA 22908 USA. RP Landers, JP (reprint author), Univ Virginia, Dept Chem, Mccormick Rd, Charlottesville, VA 22901 USA. NR 54 TC 33 Z9 33 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 NOV 1 PY 2000 VL 72 IS 21 BP 5483 EP 5492 DI 10.1021/ac0004916 PG 10 WC Chemistry, Analytical SC Chemistry GA 369XX UT WOS:000165094000056 PM 11080904 ER PT J AU Schunemann, HJ Stanulla, M Trevisan, M Aplan, PD Freudenheim, JL Muti, P AF Schunemann, HJ Stanulla, M Trevisan, M Aplan, PD Freudenheim, JL Muti, P TI Short-term storage of blood samples and DNA isolation in serum separator tubes for application in epidemiological studies and clinical research SO ANNALS OF EPIDEMIOLOGY LA English DT Article DE biological specimen bank; blood banking; DNA isolation; genetic epidemiology ID HUMAN-GENOME-PROJECT; EMERGING IMPORTANCE; RAPID METHOD; WHOLE-BLOOD; SOLID-PHASE; MLL GENE; CELLS; EXTRACTION; QUALITY; YIELD AB PURPOSE: To investigate the use of a simple DNA isolation technique for application in epidemiologic studies. To analyze systematically the potential impact of lag time between blood drawing and DNA isolation and the condition of storage of blood samples on the quantity and quality of isolated DNA in large-scale epidemiologic studies. METHODS: A modified single tube DNA isolation technique was used. DNA was isolated from samples collected from six participants and processed in triplicate: a) without delay after blood drawing; b) after blood cells were stored at 4 degreesC for 7 days; c) after blood cells were stored at -70 degreesC for 7 days; and d) after storage for 7 days at -70 degreesC with addition of lysis/digestion buffer. Polymerase chain reaction (PCR) and Southern blot analysis were performed to analyze the quality of the isolated DNA. RESULTS: The average amount of DNA isolated ranged from 27.0 to 71.1 mug/4.5 mi whole blood. Storage at 4 degreesC yielded, on the average, 20% less DNA than the samples processed without delay or after storage at -70 degreesC, although this difference was not statistically significant (p = 0.63). All four conditions studied allowed isolation of highly pure DNA suitable for genetic analyses by Southern blot analysis and polymerase chain reaction. CONLUSIONS: This pilot study suggests that storage for 7 days and at different temperatures allows isolation of high quality DNA. Using the described technique, storage of up to 7 days permits processing of large numbers of samples (50-70) in a single day, allowing for a reliable and cost efficient way of processing in various settings. Further studies are needed to investigate the influence of long-term storage of biological specimens on DNA isolation and quality. (C) 2000 Elsevier Science Inc. All rights reserved. C1 SUNY Buffalo, Sch Med & Biomed Sci, Dept Med, Buffalo, NY 14214 USA. SUNY Buffalo, Sch Med & Biomed Sci, Dept Social & Prevent Med, Buffalo, NY 14214 USA. Roswell Pk Canc Inst, Dept Mol Med, Buffalo, NY 14263 USA. Hannover Med Sch, Dept Pediat Hematol & Oncol, D-3000 Hannover, Germany. NCI, Gaithersburg, MD USA. RP SUNY Buffalo, Sch Med & Biomed Sci, Dept Med, 270 Farber Hall,3435 Main St, Buffalo, NY 14214 USA. RI Stanulla, Martin/D-2528-2010; Aplan, Peter/K-9064-2016 NR 33 TC 6 Z9 6 U1 1 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1047-2797 EI 1873-2585 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 BP 538 EP 544 DI 10.1016/S1047-2797(00)00076-4 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 383YL UT WOS:000165912200009 PM 11118934 ER PT J AU Alexander, GA Chu, KC Ho, RCS AF Alexander, GA Chu, KC Ho, RCS TI Representation of Asian Americans in clinical cancer trials SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE National Cancer Institute; clinical cancer research; clinical trials; randomized controlled trial; neoplasms; support; US Government; PHS; Asian Americans ID THERAPY; US AB PURPOSE: The objectives of this study are to analyze the accrual of Asian Americans to National Cancer Institute (NCI)-supported prevention, screening/diagnosis, and treatment trials and to determine if there is proportional ethnic representation. METHODS: Data were obtained on all participants accrued to ongoing prevention and screening/diagnosis trials and on all patients accrued to treatment trials from 1994 to mid-1998. In the analysis, the percentage of Asian Americans to the total number of trial participants is calculated. For treatment trials, participants were stratified into five age groups: 0-20 years, 21-44 years, 45-54 years, 55-64 years, and 65 or more years. RESULTS: Asian Americans represented 4.8% of subjects accrued in screening/diagnosis trials, 1.8 to 2.2% of subjects in treatment trials, and 0.9% of subjects in prevention trials. Comparison of treatment trial age groups revealed that younger Asian Americans participate significantly more in treatment trials than older Asian Americans. CONCLUSIONS: Asian American accrual in NCI-supported trials is representative of the cancer burden of Asian Americans in the United States. However, Asian Americans 65+ years are under-represented. Their full participation in cancer trials is justified. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NCI, Off Special Populat Res, NIH, Bethesda, MD 20892 USA. Univ Hawaii, Honolulu, HI 96822 USA. RP Alexander, GA (reprint author), NCI, Off Special Populat Res, NIH, 6120 Execut Blvd,Room 320,MSC 7161, Bethesda, MD 20892 USA. NR 16 TC 23 Z9 23 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S61 EP S67 DI 10.1016/S1047-2797(00)00198-8 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000008 PM 11189094 ER PT J AU Giuliano, AR Mokuau, N Hughes, C Tortolero-Luna, G Risendal, B Ho, RCS Prewitt, TE McCaskill-Stevens, WJ AF Giuliano, AR Mokuau, N Hughes, C Tortolero-Luna, G Risendal, B Ho, RCS Prewitt, TE McCaskill-Stevens, WJ TI Participation of minorities in cancer research: The influence of structural, cultural, and linguistic factors SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE ethnic minorities; cancer research; African Americans; Asian Americans; Hispanics; Native Americans; Native Hawaiians; Pacific Islanders ID BREAST-CANCER; AFRICAN-AMERICANS; CLINICAL-TRIALS; ALASKA NATIVES; HEALTH; WOMEN; PREVENTION; CARE; DETERMINANTS; RECRUITMENT AB Overall, participation rates in cancer clinical trials are very low, ranging from 3 to 20% of eligible participants. However, participation rates are especially low among the socially disadvantaged and racial/ethnic minority groups that have been historically underrepresented in clinical research. Structural factors such as study duration, treatment or intervention schedule, cost, time, followup visits, and side effects represent more of a barrier to participation among these groups compared with white, non-Hispanics. Attitudes, beliefs, perceptions, and knowledge regarding clinical research, and cultural characteristics of underrepresented minorities pose additional barriers to participation. This article focuses on the structural, cultural, and linguistic factors that affect participation in clinical cancer research for each major U.S. racial/ethnic group. Low socioeconomic status, speaking a primary language other than English, differences in communication styles, mistrust of research and the medical system, fear, embarrassment, and lack of knowledge about the origin of cancer appear to have a negative impact on clinical cancer research participation rates. Much of the information about these factors comes from studies of cancer screening because little data is available on the factors that prevent and facilitate participation of minorities in clinical cancer trials specifically. Such research is needed, and, given the heterogeneity within and between minority populations, should occur in several different geographic settings and with as many different minority subpopulations as possible. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Arizona, Arizona Canc Ctr, Tucson, AZ 85724 USA. Univ Arizona, Arizona Prevent Ctr, Tucson, AZ 85724 USA. Univ Hawaii, Sch Social Work, Honolulu, HI 96822 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Dept Hlth, Honolulu, HI USA. Univ Hawaii, Straub Clin, Honolulu, HI 96822 USA. Loyola Univ, Stritch Sch Med, Maywood, IL 60153 USA. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Giuliano, AR (reprint author), Univ Arizona, Arizona Canc Ctr, 1515 N Campbell Ave,Room 4977C, Tucson, AZ 85724 USA. NR 83 TC 149 Z9 149 U1 2 U2 8 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 1047-2797 EI 1873-2585 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S22 EP S34 DI 10.1016/S1047-2797(00)00195-2 PG 13 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000004 PM 11189089 ER PT J AU Hughes, CK Tsark, JU Kenui, CK Alexander, GA AF Hughes, CK Tsark, JU Kenui, CK Alexander, GA TI Cancer research studies in Native Hawaiians and Pacific Islanders SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE neoplasms; cancer research; clinical cancer research; Native Hawaiian; Pacific Islander; American Samoans; Chamorros; Palauans; Mariana Islanders; Marshall Islanders; Micronesians ID FALLOUT AB PURPOSE: To review and assess published findings from relevant cancer research studies in Native Hawaiians and other Pacific Islanders and to develop strategies for designing and implementing successful cancer research studies in the future. METHODS: Data were collected primarily from MEDLINE and BIOSIS Preview searches of the English literature during a 30-year period for published reports of cancer surveillance studies and epidemiological and clinical cancer studies in the Native Hawaiian and Pacific Islander populations. The cancer burden was critically assessed in the retrieved citations for each of the indigenous groups from Hawai'i, American Samoa, Guam, Commonwealth of the Northern Mariana Islands, Federated States of Micronesia, Republic of Palau, and the Republic of the Marshall Islands. RESULTS: A review of the published literature revealed a lack of systematic data collection on cancer incidence and mortality in Pacific islanders. Wide variations were found regarding the status of cancer research among ethnic groups. It is estimated that Native Hawaiians represent 0.1% of subjects accrued to cancer prevention trials, and that Pacific Islanders represent 0.5% of subjects in a large cancer screening trial. CONCLUSION: The paucity of cancer data and clinical cancer research supports the need for increased attention to these indigenous populations to improve the quality of cancer care in Native Hawaiian and Pacific Island communities. (C) 2000 Elsevier Science Inc All rights reserved. C1 Hawaii State Dept Hlth, Off Hlth Par, Honolulu, HI 96813 USA. Papa Ola Lokahi, Honolulu, HI USA. NCI, Off Special Populat Res, Bethesda, MD 20892 USA. RP Hughes, CK (reprint author), Hawaii State Dept Hlth, Off Hlth Par, 1250 Punchbowl St,Suite 210, Honolulu, HI 96813 USA. NR 39 TC 7 Z9 7 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S49 EP S60 DI 10.1016/S1047-2797(00)00193-9 PG 12 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000007 PM 11189093 ER PT J AU Moinpour, CM Atkinson, JO Thomas, SM Underwood, SM Harvey, C Parzuchowski, J Lovato, LC Ryan, AM Hill, MS DeAntoni, E Gritz, ER Thompson, IM Coltman, CA AF Moinpour, CM Atkinson, JO Thomas, SM Underwood, SM Harvey, C Parzuchowski, J Lovato, LC Ryan, AM Hill, MS DeAntoni, E Gritz, ER Thompson, IM Coltman, CA TI Minority recruitment in the prostate cancer prevention trial SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE minority recruitment; cancer prevention trials ID CLINICAL-TRIALS; PCPT UPDATE; BLACKS AB PURPOSE: African American men have a higher prostate cancer risk profile than that of other men in the United States. The purpose of this manuscript is to summarize the challenges associated with enrolling and randomizing African American and other minority participants in the Prostate Cancer Prevention Trial (PCPT). METHODS: The PCPT is a randomized trial of finasteride versus placebo for preventing prostate cancer in healthy men age 55 years and older; it is coordinated by the Southwest Oncology Group. The manuscript describes demographic and lifestyle characteristics of the PCPT randomized sample (18,882 men) by four racial and ethnic groups (Caucasian, African American, Hispanic, and other). African American men comprised 4% of the total randomized sample compared to our goal of 8%. Minority recruitment was emphasized through the Study Manual and training that occurred at trial activation. Supplemental minority recruitment activities were initiated a year after study activation and continued through the end of the accrual period. Minority recruitment was emphasized as follows: minority recruitment presentations at PCPT training seminars (held during twice yearly Southwest Oncology Group meetings); distribution of additional minority recruitment materials; engagement of four consultants for minority recruitment; production of a Minority Recruitment Manual; and a small pilot study involving minority outreach recruiters at five PCPT sites. RESULTS: The consultants were helpful in implementing the pilot project and in suggesting and reviewing materials for minority recruitment. The five-site pilot project did not increase either enrollment or randomization of minorities (with a possible exception at one site). CONCLUSIONS: We suggest that a long term perspective is required for successful recruitment of minority participants in clinical trials. Likewise, extensive minority recruitment efforts must be ready to implement at trial activation. (C) 2000 Elsevier Science Inc. All rights reserved. C1 SW Oncol Grp, Operat Off, San Antonio, TX 78245 USA. Fred Hutchinson Canc Res Ctr, SW Oncol Grp, Ctr Stat, Seattle, WA 98109 USA. NCI, NIH, Washington, DC USA. George Washington Univ, Sch Med, Washington, DC USA. Louisiana State Univ, Med Ctr, Stanley S Scott Canc Ctr, New Orleans, LA USA. Univ Wisconsin, Sch Nursing, Milwaukee, WI 53201 USA. Univ Texas, Sch Nursing, Tyler, TX USA. Wayne State Univ, Sch Med, Dept Urol, Southfield, MI USA. Hospice Michigan, Southfield, MI USA. Univ Colorado, Hlth Sci Ctr, Denver, CO USA. Univ Texas, MD Anderson Canc Ctr, Dept Behav Sci, Houston, TX 77030 USA. Univ Texas, Hlth Sci Ctr, Div Urol, San Antonio, TX USA. RP Moinpour, CM (reprint author), SW Oncol Grp, Operat Off, S9217,14980 Omicron Dr, San Antonio, TX 78245 USA. FU NCI NIH HHS [2 U10 CA37429-09, 5 U10 CA37429] NR 21 TC 48 Z9 48 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S85 EP S91 DI 10.1016/S1047-2797(00)00185-X PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000011 PM 11189097 ER PT J AU Pinto, HA McCaskill-Stevens, W Wolfe, P Marcus, AC AF Pinto, HA McCaskill-Stevens, W Wolfe, P Marcus, AC TI Physician perspectives on increasing minorities in cancer clinical trials: An Eastern Cooperative Oncology Group (ECOG) initiative SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE African American; cancer; clinical trials; focus groups; patient selection ID CARE; POPULATIONS AB PURPOSE: This paper describes the ECOG NMA Minority Accrual Initiative to assure minority participation in cancer clinical trials. METHODS: Focus groups were held to identify physician reported barriers to the enrollment of minority patients in Cleveland, OH, Indianapolis, IN, Santa Clara County, CA, and Philadelphia, PA. Community physicians affiliated with the National Medical Association (NMA), and Eastern Cooperative Oncology Group (ECOG) investigators participated in the focus groups. A four-step process consisting of focus group workshops were conducted to (i) identify barriers, (ii) develop potential solutions to the barriers, (iii) define solutions to barriers involving specific clinical trials, and (iv) implement the solutions. RESULTS: Focus group participants identified physician lack of information, patient fears and suspicion, the fear of losing patients, and distrust of the health care system as the major barriers to enrollment of African Americans. We found significant differences between community physicians and cancer program physicians in several areas. Community physicians emphasized personal contacts to address the lack of information and to overcome patient fears and suspicions, while the cancer program physicians emphasized printed materials. There was no difference by region in the barriers identified in the focus group workshops; however, the proposed solutions to overcoming the barriers were specific to each site. CONCLUSION: The four-step process developed by the ECOG and the NMA used the focus group methodology to identify and overcome barriers to participation of African Americans in cancer clinical trials. Outreach efforts to educate patients, their families, and community physicians about cancer clinical trials should be directed at overcoming patient suspicions and providing practical information to physicians about specific trials and how to enroll patients. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Stanford Univ, Sch Med, Med Ctr, Dept Med,Div Oncol, Stanford, CA 94305 USA. VA Palo Alto Hlth Care Syst, Med Serv, Oncol Sect, Palo Alto, CA USA. NCI, Div Canc Prevent, NIH, Rockville, MD USA. AMC Canc Res Ctr, Denver, CO USA. RP Pinto, HA (reprint author), Stanford Univ, Sch Med, Med Ctr, Dept Med,Div Oncol, 703 Welch Rd, Stanford, CA 94305 USA. FU NCI NIH HHS [2 U10 CA 211115-20] NR 16 TC 43 Z9 43 U1 1 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S78 EP S84 DI 10.1016/S1047-2797(00)00191-5 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000010 PM 11189096 ER PT J AU Royal, C Baffoe-Bonnie, A Kittles, R Powell, I Bennett, J Hoke, G Pettaway, C Weinrich, S Vijayakumar, S Ahaghotu, C Mason, T Johnson, E Obeikwe, M Simpson, C Mejia, R Boykin, W Roberson, P Frost, J Faison-Smith, L Meegan, C Foster, N Furbert-Harris, P Carpten, J Bailey-Wilson, J Trent, J Berg, K Dunston, G Collins, F AF Royal, C Baffoe-Bonnie, A Kittles, R Powell, I Bennett, J Hoke, G Pettaway, C Weinrich, S Vijayakumar, S Ahaghotu, C Mason, T Johnson, E Obeikwe, M Simpson, C Mejia, R Boykin, W Roberson, P Frost, J Faison-Smith, L Meegan, C Foster, N Furbert-Harris, P Carpten, J Bailey-Wilson, J Trent, J Berg, K Dunston, G Collins, F TI Recruitment experience in the first phase of the African American Hereditary Prostate Cancer (AAHPC) Study SO ANNALS OF EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT Workshop on Participation of Minorities and Women in Clinical Cancer Research CY FEB 26-27, 1999 CL WASHINGTON, D.C. SP NCI DE African American; prostate cancer; genetics ID HUMAN-GENOME-PROJECT; CLINICAL-TRIALS; SUSCEPTIBILITY LOCUS; PARTICIPATION; POPULATIONS; CHROMOSOME; PREVENTION; TUSKEGEE; SYSTEM; RISK AB The African American Hereditary Prostate Cancer (AAHPC) Study is an ongoing multicenter genetic linkage study organized by Howard University and the National Human Genome Research Institute (NHGRI), with support from the Office for Research on Minority Health and the National Cancer institute. The goals of the study are to: (i) look for evidence of involvement of chromosome 1q24-25 (HPC1) in African American men with hereditary prostate cancer (HPC) and (ii) conduct a genome wide search for other loci associated with HPC in African American men. To accomplish these goals, a network has been established including Howard University, the NHGRI, and sh Collaborative Recruitment Centers (CRCs). The CRCs are responsible for the identification and enrollment of 100 African American families. To date, 43 families have been enrolled. Recruitment strategies have included mass media campaigns, physician referrals, community health-fairs/prostate cancer screenings, support groups, tumor registries, as well as visits to churches, barber shops, and universities. By far, the most productive recruitment mechanisms have been physician referrals and tumor registries, yielding a total of 35 (81%) families. Approximately 41% (n = 3400) of probands initially contacted by phone or mail expressed interest in participating; the families of 2% of these mel the eligibility criteria, and 75% of those families have been enrolled in the study, indicating a 0.5% recruitment yield (ratio of participants to contacts). As the first large-scale genetic linkage study of African Americans, on a common disease, the challenges and successes of the recruitment process for the AAHPC Study should serve to inform future efforts to involve this population in similar studies. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Howard Univ, Natl Human Genome Ctr, Washington, DC 20059 USA. NHGRI, NIH, Bethesda, MD USA. Fox Chase Canc Ctr, Philadelphia, PA 19111 USA. Wayne State Univ, Detroit, MI USA. Midtown Urol, Atlanta, GA USA. Columbia Presbyterian Med Ctr, New York, NY 10032 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Univ S Carolina, Columbia, SC 29208 USA. Univ Chicago Univ Illinois, Ctr Radiat Therapy, Michael Reese Hosp, Chicago, IL USA. Univ Illinois, Chicago, IL USA. RP Royal, C (reprint author), Howard Univ, Natl Human Genome Ctr, 2216 6th St NW,Room 207, Washington, DC 20059 USA. FU NHGRI NIH HHS [N01-HG-75418] NR 48 TC 63 Z9 63 U1 0 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S68 EP S77 DI 10.1016/S1047-2797(00)00194-0 PG 10 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000009 PM 11189095 ER PT J AU Underwood, SM Alexander, GA AF Underwood, SM Alexander, GA TI Participation of minorities and women in clinical cancer research SO ANNALS OF EPIDEMIOLOGY LA English DT Editorial Material C1 Univ Wisconsin, Sch Nursing, Milwaukee, WI 53201 USA. NCI, Bethesda, MD 20892 USA. RP Underwood, SM (reprint author), Univ Wisconsin, Sch Nursing, Milwaukee, WI 53201 USA. NR 0 TC 7 Z9 7 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1047-2797 J9 ANN EPIDEMIOL JI Ann. Epidemiol. PD NOV PY 2000 VL 10 IS 8 SU S BP S1 EP S2 DI 10.1016/S1047-2797(00)00201-5 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 384MX UT WOS:000165950000001 ER PT J AU Warach, S Pettigrew, LC Dashe, JF Pullicino, P Lefkowitz, DM Sabounjian, L Harnett, K Schwiderski, U Gammans, R AF Warach, S Pettigrew, LC Dashe, JF Pullicino, P Lefkowitz, DM Sabounjian, L Harnett, K Schwiderski, U Gammans, R CA Citicoline 010 Investigators TI Effect of citicoline on ischemic lesions as measured by diffusion-weighted magnetic resonance imaging SO ANNALS OF NEUROLOGY LA English DT Article ID EXPERIMENTAL FOCAL ISCHEMIA; CEREBRAL-ARTERY OCCLUSION; FIBROBLAST-GROWTH-FACTOR; ACUTE HUMAN STROKE; CDP-CHOLINE; EMBOLIC STROKE; PERFUSION MRI; THROMBOLYTIC THERAPY; IN-VIVO; BRAIN AB We examined the effect of the neuroprotective and neuroreparative agent citicoline on the growth of cerebral ischemic lesions in a double-blind placebo-controlled study involving patients with acute ischemic stroke using diffusion-weighted magnetic resonance imaging (DWI). Patients with acute ischemic stroke symptom onset 24 hours or less before the start of treatment, National Institutes of Health Stroke Scale (NIHSS) scores of 5 or higher, and lesions of 1 to 120 cc in cerebral gray matter by DWI were enrolled. DWI, T2-weighted magnetic resonance imaging (MRI), perfusion-weighted MRI, and magnetic resonance angiography were obtained at baseline, week 1, and week 12. Citicoline (500 mg/day) was administered orally for 6 weeks, and patients were followed for 12 weeks. The primary assessment was progression of ischemic lesion volume from baseline to 12 weeks as measured by MRI. A total of 100 patients entered the study. The primary MRI analysis included 40 placebo-treated patients and 41 citicoline-treated patients with both baseline and week 12 MRT data and failed to demonstrate a significant difference in lesion volume change from baseline to week 12. From baseline to week 12, ischemic lesion volume fall values mean (SE)I expanded by 180% (107) among placebo-treated patients compared with 34% (19) among citicoline-treated patients. In a secondary analysis, Lesion volume decreased from week 1 to week 12 by 6.9 cc (2.8) on placebo versus 17.2 cc (2.6) on citicoline. Baseline variables that were predictors of change in lesion size over 12 weeks were the volume of hypoperfusion (strongest association), baseline NIHSS score, lesion volume on DWI, arterial lesion by magnetic resonance angiography, and categorized elapsed time (less than or equal to 12 or >12 hours) from stroke onset to first dose. A marked association between lesion volume reduction and improvement of NIHSS score by seven or more points was observed. Significant correlations between lesion volumes and clinical measures were found, replicating values reported in the literature for smaller case series. We observed a reduction in lesion volume growth from baseline to week 12 with citicoline treatment, with a significantly greater reduction in volume from week 1 to week 12 with citicoline. We found a significant inverse relationship between lesion volume change over 12 weeks as measured by MRI and clinical outcome for ischemic stroke. This relationship supports the role of DWI as a surrogate marker of clinically meaningful lesion progression in stroke clinical trials. The hypothesis that citicoline reduces lesion growth and improves clinical outcome will be tested further. C1 NINDS, NIH, Bethesda, MD 20892 USA. RP Warach, S (reprint author), NINCDS, NIH, 36 Convent Dr,MSC 4129,Room 4A03, Bethesda, MD 20892 USA. NR 60 TC 135 Z9 140 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD NOV PY 2000 VL 48 IS 5 BP 713 EP 722 DI 10.1002/1531-8249(200011)48:5<713::AID-ANA4>3.0.CO;2-# PG 10 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 370NW UT WOS:000165130000004 PM 11079534 ER PT J AU Sullivan, PG Geiger, JD Mattson, MP Scheff, SW AF Sullivan, PG Geiger, JD Mattson, MP Scheff, SW TI Dietary supplement creatine protects against traumatic brain injury SO ANNALS OF NEUROLOGY LA English DT Article ID MITOCHONDRIAL DYSFUNCTION; CYCLOSPORINE-A; PERMEABILITY TRANSITION; CORTICAL CONTUSION; ERGOGENIC AID; HEAD-INJURY; DAMAGE; KINASE; RAT; CYCLOCREATINE AB Creatine, one of the most common food supplements used by individuals at almost every level of athleticism, promote gains in performance, strength, and fat-free mass. Recent experimental findings have demonstrated that creatine affords significant neuroprotection against ischemic and oxidative insults. The present experiments investigated the possible effect of creatine dietary supplementation on brain tissue damage after experimental traumatic brain injury. Results demonstrate that chronic administration of creatine ameliorated the extent of cortical damage by as much as 36% in mice and 50% in rats. Protection seems to be related to creatine-induced maintenance of mitochondrial bioenergetics. Mitochondrial membrane potential was significantly increased, intramitochondrial levels of reactive oxygen species and calcium were significantly decreased, and adenosine triphosphate levels were maintained. Induction of mitochondrial permeability transition was significantly inhibited in animals fed creatine. This food supplement may provide clues to the mechanisms responsible for neuronal loss after traumatic brain injury and may find use as a neuroprotective agent against acute and delayed neurodegenerative processes. C1 Univ Kentucky, Sanders Brown Ctr Aging 233, Lexington, KY 40536 USA. Univ Manitoba, Dept Pharmacol & Therapeut, Winnipeg, MB, Canada. NIH, Neurosci Lab, Baltimore, MD USA. RP Scheff, SW (reprint author), Univ Kentucky, Sanders Brown Ctr Aging 233, 800 S Limestone Ave, Lexington, KY 40536 USA. RI Mattson, Mark/F-6038-2012 FU NHLBI NIH HHS [HL60287]; NINDS NIH HHS [NS31220, NS39828] NR 49 TC 164 Z9 173 U1 1 U2 13 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD NOV PY 2000 VL 48 IS 5 BP 723 EP 729 DI 10.1002/1531-8249(200011)48:5<723::AID-ANA5>3.0.CO;2-W PG 7 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 370NW UT WOS:000165130000005 PM 11079535 ER PT J AU Chan, FKM AF Chan, FKM TI The pre-ligand binding assembly domain: a potential target of inhibition of tumour necrosis factor receptor function SO ANNALS OF THE RHEUMATIC DISEASES LA English DT Article ID AUTOIMMUNE LYMPHOPROLIFERATIVE SYNDROME; FACTOR TNF RECEPTOR; FAS GENE-MUTATIONS; NF-KAPPA-B; LYMPHOCYTE APOPTOSIS; DEPENDENT APOPTOSIS; CRYSTAL-STRUCTURE; ENERGY-TRANSFER; CELL-SURFACES; DEATH DOMAIN AB Signalling by the tumour necrosis factor receptors (TNFR) is thought to be mediated by the binding of the trimeric Ligand TNF to three monomeric subunits of the receptor. This Ligand induced trimerisation model of TNFR signalling is mainly supported by crystallographic data of the p60 TNFR-1 and TNF beta complex in which the trimeric Ligand interdigitates between the individual receptor chains and prevents the receptor subunits from interacting with each other. Recently, a domain NH, terminal to the ligand binding domain in the extracellular region of p60 TNFR-1, p80 TNFR-2 and Fas was identified that mediates receptor self association before Ligand binding. This pre-ligand binding assembly domain or PLAD is critical for assembly of functional receptor complexes on the cell surface and may provide a potential target in the design of future novel therapeutics against diseases mediated by members of the TNFR family of receptors. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Chan, FKM (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Room 11N311, Bethesda, MD 20892 USA. RI Chan, Francis/E-9647-2014 OI Chan, Francis/0000-0002-4803-8353 NR 44 TC 19 Z9 20 U1 0 U2 1 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0003-4967 J9 ANN RHEUM DIS JI Ann. Rheum. Dis. PD NOV PY 2000 VL 59 SU 1 BP 50 EP 53 PG 4 WC Rheumatology SC Rheumatology GA 375NV UT WOS:000165408900014 ER PT J AU Morris, JC Waldmann, TA AF Morris, JC Waldmann, TA TI Advances in interleukin 2 receptor targeted treatment SO ANNALS OF THE RHEUMATIC DISEASES LA English DT Article ID VERSUS-HOST DISEASE; T-CELL LEUKEMIA; SYSTEMIC LUPUS-ERYTHEMATOSUS; BONE-MARROW TRANSPLANTATION; HUMANIZED ANTI-TAC; JAK-3 JANUS KINASE; RHEUMATOID-ARTHRITIS; MONOCLONAL-ANTIBODY; IL-2 RECEPTOR; SERUM LEVELS AB T cell activation and cellular immune responses are modulated by interleukin 2 (IL2) through binding to its corresponding cell surface receptor. Three forms of the receptor are recognised based on IL2 binding affinity. The high affinity receptor is a heterotrimer composed of alpha, beta, and gamma (c)-polypeptide chains. The 55 kDa alpha -chain also known as the Tac (T cell activation) antigen or CD-25 is a unique subunit of the high affinity IL2 receptor (IL2R alpha). Resting T cells express few IL2R alpha, however, when activated, the expression of ILR2 alpha rapidly increases. The IL2Ra is shed from the cell surface and is measurable in the serum as a 45 kDa soluble form (s-Tac or s-IL2R alpha). Serum concentrations of s-Tac can be used as a surrogate marker for T cell activation and IL2Ra expression. IL2Ra is over expressed by T cells in a number of autoimmune diseases, allograft rejection and a variety of lymphoid neoplasms. IL2 induced proliferation of T cells can be inhibited by the murine monoclonal antibody (anti-Tac) directed against the alpha -chain of the IL2R. Through molecular engineering, murine anti-Tac has been humanised reducing its immunogenicity without changing its specificity. Humanised anti-Tac (HAT) has been shown to reduce the incidence of renal and cardiac allograft rejection as well as decrease the severity of graft versus host disease in patients undergoing HLA matched allogeneic bone marrow transplantation. IL2Ra targeted treatment with radioimmunoconjugates of anti-Tac and immunotoxins has shown promise in the treatment of CD25 expressing lymphomas. C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Morris, JC (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, Bldg 10,Rm 4N115,10 Ctr Dr, Bethesda, MD 20892 USA. NR 85 TC 74 Z9 80 U1 0 U2 2 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0003-4967 J9 ANN RHEUM DIS JI Ann. Rheum. Dis. PD NOV PY 2000 VL 59 SU 1 BP 109 EP 114 PG 6 WC Rheumatology SC Rheumatology GA 375NV UT WOS:000165408900025 ER PT J AU O'Shea, JJ Visconti, R Cheng, TP Gadina, M AF O'Shea, JJ Visconti, R Cheng, TP Gadina, M TI Jaks and Stats as therapeutic targets SO ANNALS OF THE RHEUMATIC DISEASES LA English DT Article ID SEVERE COMBINED IMMUNODEFICIENCY; TYROSINE KINASE DOMAIN; MICE LACKING JAK3; TRANSCRIPTIONAL ACTIVATION; JANUS KINASE-3; LYMPHOID DEVELOPMENT; SIGNAL-TRANSDUCTION; CYTOKINE RECEPTORS; MAP KINASE; TH2 CELLS AB Cytokines have critical functions in regulating immune responses. A large number of these factors bind related receptors termed the Type I and Type II families of cytokine receptors. These receptors activate Janus kinases (Jaks) and Stat family of transcription factors. The essential and specific function of Jaks and Stats is particularly well illustrated by human and mouse mutations. The possibility that these molecules could be targeted to produce novel immunosuppressive compounds is considered in this review. C1 NIH, Bethesda, MD 20892 USA. NIAMSD, Lymphotcyte Cell Biol Sect, Bethesda, MD 20892 USA. NIAMSD, Rheumatism Branch, Bethesda, MD 20892 USA. RP O'Shea, JJ (reprint author), NIH, Bldg 10,Room 9N252,10 ctr Dr MSC-1820, Bethesda, MD 20892 USA. RI Visconti, Roberta/C-5299-2009 NR 62 TC 10 Z9 10 U1 0 U2 2 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0003-4967 J9 ANN RHEUM DIS JI Ann. Rheum. Dis. PD NOV PY 2000 VL 59 SU 1 BP 115 EP 118 PG 4 WC Rheumatology SC Rheumatology GA 375NV UT WOS:000165408900026 ER PT J AU Kakeya, H Miyazaki, Y Miyazaki, H Nyswaner, K Grimberg, B Bennett, JE AF Kakeya, H Miyazaki, Y Miyazaki, H Nyswaner, K Grimberg, B Bennett, JE TI Genetic analysis of azole resistance in the Darlington strain of Candida albicans SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID AMINO-ACID SUBSTITUTIONS; LANOSTEROL 14-ALPHA-DEMETHYLASE; FLUCONAZOLE RESISTANCE; CONTRIBUTE; CORRELATE; MUTATION; DEMETHYLASE; SEQUENCE; AFFINITY AB High-level azole resistance in the Darlington strain of Candida albicans was investigated by gene replacement in C. albicans and expression in Saccharomyces cerevisiae, We sequenced the ERG11 gene, which encodes the sterol C(14)alpha -demethylase, from our copy of the Darlington strain. Both alleles contained the histidine for tyrosine substitution at position 132 (Y132H) reported in Darlington by others, but we also found a threonine-for-isoleucine substitution (I471T) not previously reported in the C, albicans ERG11. The encoded I471T change in amino acids conferred azole resistance when overexpressed alone and increased azole resistance when added to the Y132H amino acid sequence in an S. cerevisiae expression system, Replacement of one copy of ERG11 in an azole-susceptible strain of C, albicans with a single copy of the Darlington ERG11 resulted in expression of the integrated copy and a modest increase in azole resistance. The profound azole resistance of the Darlington strain is the result of multiple mutations. C1 NIAID, Clin Mycol Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Nagasaki Univ, Sch Med, Dept Internal Med 2, Nagasaki 8528061, Japan. RP Bennett, JE (reprint author), NIH, Ctr Clin, Rm 11C304, Bethesda, MD 20892 USA. RI Grimberg, Brian/I-1251-2013 OI Grimberg, Brian/0000-0002-6015-4912 NR 26 TC 50 Z9 61 U1 0 U2 2 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 NOV PY 2000 VL 44 IS 11 BP 2985 EP 2990 DI 10.1128/AAC.44.11.2985-2990.2000 PG 6 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 366WL UT WOS:000090029200009 PM 11036010 ER PT J AU Ma, L Kovacs, JA AF Ma, L Kovacs, JA TI Expression and characterization of recombinant human-derived Pneumocystis carinii dihydrofolate reductase SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID TRIMETHOPRIM-SULFAMETHOXAZOLE; DIHYDROPTEROATE SYNTHASE; TOXOPLASMA-GONDII; PNEUMONIA; INHIBITORS; MUTATIONS; GENE; PROPHYLAXIS; BINDING; THERAPY AB Dihydrofolate reductase (DHFR) is the target of trimethoprim (TMP), which has been widely used in combination with sulfa drugs for treatment and prophylaxis of Pneumocystis carinii pneumonia, While the rat-derived P. carinii DHFR has been well characterized, kinetic studies of human-derived P, carinii DHFR, which differs from rat-derived P. carinii DHFR by 38% in amino acid sequence, have not been reported to date. Here we report on the expression and kinetic characterization of the recombinant human-derived P, carinii DHFR The 618-bp coding sequence of the human-derived P, carinii DHFR gene was expressed in Escherichia call, As determined by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis, the purified enzyme had a molecular mass of 25 kDa, consistent with that predicted from the DNA sequence. Kinetic analysis showed that the K-m values for dihydrofolate and NADPH were 2.7 +/- 0.3 and 14.0 +/- 4.3 muM, respectively, which are similar to those reported for rat-derived P. carinii DHFR. Inhibition studies revealed that both TMP and pyrimethamine were poor inhibitors of human-derived P, carinii DHFR, with K-i values of 0.28 +/- 0.08 and 0.065 +/- 0.005 muM, respectively, while trimetrexate and methotrexate were potent inhibitors, with K-i values of 0.23 +/- 0.03 and 0.016 +/- 0.004 nM, respectively. The availability of purified recombinant enzyme in large quantities should facilitate the identification of antifolate inhibitors with greater potency and higher selectivity for human-derived P. carinii DHFR. C1 NIH, Warren Grant Magnuson Clin Ctr, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Kovacs, JA (reprint author), NIH, Warren Grant Magnuson Clin Ctr, Dept Crit Care Med, Bldg 10,Room 7D43,10 Ctr Dr,MSC 1662, Bethesda, MD 20892 USA. NR 27 TC 21 Z9 21 U1 0 U2 3 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 NOV PY 2000 VL 44 IS 11 BP 3092 EP 3096 DI 10.1128/AAC.44.11.3092-3096.2000 PG 5 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 366WL UT WOS:000090029200027 PM 11036028 ER PT J AU Yan, SS Fox, ML Holland, SM Stock, F Gill, VJ Fedorko, DP AF Yan, SS Fox, ML Holland, SM Stock, F Gill, VJ Fedorko, DP TI Resistance to multiple fluoroquinolones in a clinical isolate of Streptococcus pyogenes: Identification of gyrA and parC and specification of point mutations associated with resistance SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID IN-VITRO ACTIVITIES; DNA GYRASE; TOPOISOMERASE-IV; PNEUMONIAE; SPARFLOXACIN; QUINOLONES; INFECTIONS; SUBUNIT; TARGETS; GENE AB A strain of Streptococcus pyogenes resistant to multiple fluoroquinolones was isolated from the blood of an immunocompromised patient. Resistance to fluoroquinolones in S. pyogenes has not been previously studied. Compared to 10 sensitive strains of S. pyogenes, the fluoroquinolone-resistant clinical isolate of S, pyogenes presented point mutations in gyrA, predicting that serine-81 was changed to phenylalanine and that methionine-99 was changed to leucine, and in parC, predicting that serine-79 was changed to tyrosine. The mechanism of fluoroquinolone resistance in this isolate of S. pyogenes appears to be analogous to previously reported mechanisms for Streptococcus pneumoniae. C1 NIAID, Microbiol Serv, CPD, Ctr Clin,NIH, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Fedorko, DP (reprint author), NIAID, Microbiol Serv, CPD, Ctr Clin,NIH, 9000 Rockville Pike,Bldg 10,Rm 2C385, Bethesda, MD 20892 USA. NR 18 TC 47 Z9 49 U1 0 U2 2 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 NOV PY 2000 VL 44 IS 11 BP 3196 EP 3198 DI 10.1128/AAC.44.11.3196-3198.2000 PG 3 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 366WL UT WOS:000090029200051 PM 11036052 ER PT J AU Stevenson, DK Verter, J Fanaroff, AA Oh, W Ehrenkranz, RA Shankaran, S Donovan, EF Wright, LL Lemons, JA Tyson, JE Korones, SB Bauer, CR Stoll, BJ AF Stevenson, DK Verter, J Fanaroff, AA Oh, W Ehrenkranz, RA Shankaran, S Donovan, EF Wright, LL Lemons, JA Tyson, JE Korones, SB Bauer, CR Stoll, BJ CA L-A Papile Natl I Child Hlth Human TI Sex differences in outcomes of very low birthweight infants: the newborn male disadvantage SO ARCHIVES OF DISEASE IN CHILDHOOD LA English DT Article DE very low birthweight infants; neonatal outcome; sex; mortality ID BIRTH-WEIGHT OUTCOMES; NEONATAL-MORTALITY; RESOURCE USE; MORBIDITY; NETWORK; GENDER AB Objective-To determine the differences in short term outcome of very low birthweight infants attributable to sex. Methods-Boys and girls weighing 501-1500 g admitted to the 12 centres of the National Institute of Child Health and Human Development Neonatal Research Network were compared. Maternal information and perinatal data were collected from hospital records. Infant outcome was recorded at discharge, at 120 days of age if the infant was still in hospital, or at death. Best obstetric estimate based on the last menstrual period, standard obstetric factors, and ultrasound were used to assign gestational age in completed weeks. Data were collected on a cohort that included 3356 boys and 3382 girls, representing all inborn births from 1 May 1991 to 31 December 1993. Results-Mortality for boys was 22% and that for girls 15%. The prenatal and perinatal data indicate few differences between the sex groups, except that boys were less Likely to have been exposed to antenatal steroids (odds ratio (OR) = 0.80) and were less stable after birth, as reflected in a higher percentage with lower Apgar scores at one and five minutes and the need for physical and pharmacological assistance. In particular, boys were more likely to have been intubated (OR = 1.16) and to have received resuscitation medication (OR=1.40). Boys had a higher risk (OR > 1.00) for most adverse neonatal outcomes. Although pulmonary morbidity predominated, intracranial haemorrhage and urinary tract infection were also more common. Conclusions-Relative differences in short term morbidity and mortality persist between the sexes. C1 Stanford Univ, Sch Med, Dept Pediat, Stanford, CA 94305 USA. George Washington Univ, Ctr Biostat, Washington, DC USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. Brown Univ, Women & Infants Hosp, Providence, RI USA. Yale Univ, New Haven, CT USA. Wayne State Univ, Detroit, MI USA. Univ Cincinnati, Cincinnati, OH USA. NICHHD, Bethesda, MD 20892 USA. Indiana Univ, Indianapolis, IN 46204 USA. Univ Texas, SW Med Ctr, Dallas, TX USA. Univ Tennessee, Memphis, TN USA. Univ Miami, Miami, FL 33152 USA. Emory Univ, Atlanta, GA 30322 USA. Univ New Mexico, Albuquerque, NM 87131 USA. RP Stevenson, DK (reprint author), Stanford Univ, Sch Med, Dept Pediat, Stanford, CA 94305 USA. FU NICHD NIH HHS [U01 HD19897, U10 HD21364, U10 HD27880] NR 11 TC 155 Z9 159 U1 0 U2 8 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 NOV PY 2000 VL 83 IS 3 SI SI BP F182 EP F185 DI 10.1136/fn.83.3.F182 PG 4 WC Pediatrics SC Pediatrics GA 370ME UT WOS:000165126200005 PM 11040165 ER PT J AU Bohannon, AD Cooper, GS Wolff, MS Meier, DE AF Bohannon, AD Cooper, GS Wolff, MS Meier, DE TI Exposure to 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene(DDT) in relation to bone mineral density and rate of bone loss in menopausal women SO ARCHIVES OF ENVIRONMENTAL HEALTH LA English DT Article DE bone mineral density; DDE; DDT; environmental estrogens; race ID BREAST-CANCER; POSTMENOPAUSAL WOMEN; RACIAL-DIFFERENCES; PREMENOPAUSAL; WHITE; RISK; ASSOCIATION; ANDROGENS; TAMOXIFEN; ESTROGEN AB The organochlorine pesticide 2,2-bis(p-chlorophenyl)-1,1,1 -trichloroethane (DDT) and its metabolite 1,1 -dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) are examples of an environmental contaminant that may have hormonal properties. Bone metabolism is both estrogen- and androgen-dependent. Exposures to various environmental endocrine disrupters can affect bone metabolism in animals, but there are no published data concerning the effect of DDE exposure on bone metabolism in humans. We hypothesized that high levels of DDE would be associated with lower bone density in peri- and postmenopausal women than in premenopausal women. Study subjects were drawn from the cohort of women who had participated in the Mount Sinai Medical Center Longitudinal Normative Bone Density Study (1984-1987). We used serum samples obtained at study entry to measure DDE levels in 103 (50 black, 53 white) women (mean age = 54.5 y [standard deviation = 5 yl). Measurements of bone mineral density at the lumbar spine and radius were made at 6-mo intervals during a 2-y period. DDE concentrations were significantly (p <.001) higher in blacks (13.9 ng/ml) than in whites (8.4 ng/ml), but there was no correlation between DDE concentration and bone density at the spine (mean levels = 1.065 g/cm(2) and 1.043 g/cm(2) in the lowest and highest quartiles, respectively, of DDE [trend p value =.85]) or at the radius (mean levels = 0.658 g/cm and 0.664 g/cm in the lowest and highest quartiles, respectively, of DDE [trend p value =.34]). Longitudinal analyses revealed no correlation between DDE and the rate of bone loss at either bone site. Similar results were seen in race-stratified analyses, as well as in analyses in which we controlled for lactation history and other potential confounders. We found little evidence that chronic low-level DDT exposure is associated with bone density in peri- and postmenopausal women. C1 NIEHS, Res Triangle Pk, NC 27709 USA. Duke Univ, Med Ctr, Durham, NC USA. CUNY Mt Sinai Sch Med, New York, NY 10029 USA. RP Bohannon, AD (reprint author), Procter & Gamble Pharmaceut, N Amer Med Affairs, Box 2099,8700 Mason Montgomery Rd, Mason, OH 45040 USA. FU NCRR NIH HHS [M01-RR00030]; NIA NIH HHS [5P60AG11268]; NIEHS NIH HHS [1-ES 35357] NR 25 TC 11 Z9 12 U1 0 U2 2 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 NOV-DEC PY 2000 VL 55 IS 6 BP 386 EP 391 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health GA 378LF UT WOS:000165585300003 PM 11128875 ER PT J AU Kunert, KS Tisdale, AS Stern, ME Smith, JA Gipson, IK AF Kunert, KS Tisdale, AS Stern, ME Smith, JA Gipson, IK TI Analysis of topical cyclosporine treatment of patients with dry eye syndrome - Effect on conjunctival lymphocytes SO ARCHIVES OF OPHTHALMOLOGY LA English DT Article ID SECONDARY SJOGRENS-SYNDROME; KERATOCONJUNCTIVITIS SICCA; LACRIMAL GLANDS; OCULAR SURFACE; EXPRESSION; ADHESION; CELLS; INFLAMMATION; EPITHELIUM; SPECIMENS AB Objective: To study the effect of topical cyclosporine on lymphocyte activation within the conjunctiva of patients with moderate to severe dry eye syndrome (Sjogren and non-Sjogren). Methods: Biopsy specimens were obtained at baseline and after 6 months of cyclosporine treatment from eyes of 32 patients with moderate to severe dry eye syndrome; 19 were cyclosporine treated (0.05% cyclosporine, n=13; 0.1% cyclosporine, n=6) and 13 were vehicle treated. Within this group there were 12 with Sjogren syndrome and 20 with non-Sjogren syndrome. Biopsy tissue was analyzed using immunohistochemical localization of binding of monoclonal antibodies to lymphocytic markers CD3, CD4, and CD8 as well as lymphocyte activation markers CD11a and HLA-DR. Results: In cyclosporine-treated eyes, biopsy results of conjunctivae showed decreases in the number of cells positive for CD3, CD4, and CD8, while in vehicle-treated eyes, results showed increases in these markers, although these differences were not statistically significant. Following treatment with 0.05% cyclosporine, there was a significant decrease in the number of cells expressing the lymphocyte activation markers CD 11a (P<.05) and HLA-DR (P<.05), indicating less activation of lymphocytes as compared with vehicle treatment. Within the Sjogren patient subgroup, those treated with 0.05% cyclosporine also showed a significant decrease in the number of cells positive for CD11a (P<.001) as well as CD3 (P<.03), indicating a reduction in number of activated lymphocytes. Conclusion: Treatment of dry eye syndrome with topical cyclosporine significantly reduced the numbers of activated lymphocytes within the conjunctiva. C1 Schepens Eye Res Inst, Boston, MA 02114 USA. Harvard Univ, Sch Med, Dept Ophthalmol, Boston, MA USA. NEI, Bethesda, MD 20892 USA. Allergan Inc, Irvine, CA 92623 USA. RP Gipson, IK (reprint author), Schepens Eye Res Inst, Boston, MA 02114 USA. NR 38 TC 158 Z9 164 U1 0 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9950 J9 ARCH OPHTHALMOL-CHIC JI Arch. Ophthalmol. PD NOV PY 2000 VL 118 IS 11 BP 1489 EP 1496 PG 8 WC Ophthalmology SC Ophthalmology GA 372KR UT WOS:000165233300001 PM 11074805 ER PT J AU Matsui, K Riemenschneider, WK Hilbert, SL Yu, ZX Takeda, K Travis, WD Moss, J Ferrans, VJ AF Matsui, K Riemenschneider, WK Hilbert, SL Yu, ZX Takeda, K Travis, WD Moss, J Ferrans, VJ TI Hyperplasia of type II pneumocytes in pulmonary lymphangioleiomyomatosis - Immunohistochemical and electron microscopic study SO ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE LA English DT Article ID THYROID TRANSCRIPTION FACTOR-1; KERATINOCYTE GROWTH-FACTOR; TRANSBRONCHIAL BIOPSY; MONOCLONAL-ANTIBODIES; LUNG; LYMPHANGIOMYOMATOSIS; EXPRESSION; EMPHYSEMA; DIAGNOSIS; ANTIGEN AB Background.-Little is known of the morphology of the pneumocytes lining the parenchymal cysts in lymphangioleiomyomatosis (LAM), Objective.-To evaluate the structural characteristics of the alveolar epithelial cells in LAM. Methods-Immunohistochemical and electron microscopic studies were performed on lung tissue from 22 women with pulmonary LAM. Results.-Epithelial cells that reacted with PE-10 (a mouse monoclonal antibody that recognizes the surfactant apoprotein A in type II pneumocytes) and TTF-1 tan antibody that identifies nuclear transcription factor found in type II pneumocytes) were the predominant cell type lining the surfaces of lesions of LAM and normal areas of lung. Scanning and transmission electron microscopic studies confirmed that these cells were type II pneumocytes as demonstrated by (1) apical microvilli, (2) electron-dense lamellar bodies, and (3) cytoplasmic projections that extended from the basal surfaces into the underlying connective tissue, where they made extensive contact with interstitial connective tissue cells. A few cells had morphologic characteristics of type I pneumocytes, including large flat surfaces lacking microvilli. Cells that appeared intermediate between type I and type II pneumocytes were observed occasionally. Conclusions,These observations and the reactivity of these cells with antibody to proliferating cell nuclear antigen demonstrate that extensive hyperplasia of type II pneumocytes is a major characteristic of LAM. C1 NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Devices & Radiol Hlth, Rockville, MD 20857 USA. Armed Forces Inst Pathol, Dept Pulm & Mediastinal Pathol, Washington, DC 20306 USA. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. RP Ferrans, VJ (reprint author), NHLBI, Pathol Sect, NIH, Bldg 10-2N240,10 Ctr Dr MSC-1518, Bethesda, MD 20892 USA. NR 39 TC 23 Z9 24 U1 0 U2 0 PU COLLEGE AMER PATHOLOGISTS PI NORTHFIELD PA C/O KIMBERLY GACKI, 325 WAUKEGAN RD, NORTHFIELD, IL 60093-2750 USA SN 0003-9985 J9 ARCH PATHOL LAB MED JI Arch. Pathol. Lab. Med. PD NOV PY 2000 VL 124 IS 11 BP 1642 EP 1648 PG 7 WC Medical Laboratory Technology; Medicine, Research & Experimental; Pathology SC Medical Laboratory Technology; Research & Experimental Medicine; Pathology GA 374KQ UT WOS:000165344700016 PM 11079017 ER PT J AU Marcelo, A Fontelo, P Farolan, M Cualing, H AF Marcelo, A Fontelo, P Farolan, M Cualing, H TI Effect of image compression on telepathology - A randomized clinical trial SO ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE LA English DT Article; Proceedings Paper CT Meeting on Advancing Pathology Informatics, Imaging, and the Internet (APIII 99) CY OCT 14-16, 1999 CL PITTSBURGH, PENNSYLVANIA SP Univ Pittsburgh Sch Med ID VIRTUAL MICROSCOPY; JPEG AB Context-For practitioners deploying store-and-forward telepathology systems, optimization methods such as image compression need to be studied. Objective-To determine if Joint Photographic Expert Group OPC or JPEG) compression, a lossy image compression algorithm, negatively affects the accuracy of diagnosis in telepathology. Design-Double-blind randomized, controlled trial. Setting.-University-based pathology departments. Participants.-Resident and staff pathologists at the University of Illinois, Chicago, and University of Cincinnati, Cincinnati, Ohio. Intervention.-Compression of raw images using the JPEG algorithm. Main Outcome Measures.-Image acceptability, accuracy of diagnosis, confidence level of pathologist, image quality. Results.-There was no statistically significant difference in the diagnostic accuracy between noncompressed (bit map) and compressed (JPG) images. There were also no differences in the acceptability, confidence level, and perception of image quality. Additionally, rater experience did not significantly correlate with degree of accuracy. Conclusions.-For providers practicing telepathology, JPG image compression does not negatively affect the accuracy and confidence level of diagnosis. The acceptability and quality of images were also not affected. C1 Natl Lib Med, Bethesda, MD 20894 USA. Univ Illinois, Dept Pathol, Chicago, IL USA. Univ Cincinnati, Dept Pathol, Cincinnati, OH USA. RP Marcelo, A (reprint author), Natl Lib Med, 8600 Rockville Pike,38A,B1N30, Bethesda, MD 20894 USA. NR 17 TC 20 Z9 21 U1 0 U2 0 PU COLLEGE AMER PATHOLOGISTS PI NORTHFIELD PA C/O KIMBERLY GACKI, 325 WAUKEGAN RD, NORTHFIELD, IL 60093-2750 USA SN 0003-9985 J9 ARCH PATHOL LAB MED JI Arch. Pathol. Lab. Med. PD NOV PY 2000 VL 124 IS 11 BP 1653 EP 1656 PG 4 WC Medical Laboratory Technology; Medicine, Research & Experimental; Pathology SC Medical Laboratory Technology; Research & Experimental Medicine; Pathology GA 374KQ UT WOS:000165344700018 PM 11079019 ER PT J AU Wicke, C Halliday, B Allen, D Roche, NS Scheuenstuhl, H Spencer, MM Roberts, AB Hunt, TK AF Wicke, C Halliday, B Allen, D Roche, NS Scheuenstuhl, H Spencer, MM Roberts, AB Hunt, TK TI Effects of steroids and retinoids on wound healing SO ARCHIVES OF SURGERY LA English DT Article ID TRANSFORMING GROWTH-FACTOR; FACTOR-BETA; VITAMIN-A; 9-CIS-RETINOIC ACID; TGF-BETA; CELLS; RAT; AUTOCRINE; FACTOR-BETA-1; EXPRESSION AB Hypothesis: Anti-inflammatory corticosteroids significantly impair wound healing. Retinoids partially, but significantly, reverse this effect. Little is known about the mechanism of steroid retardation or retinoid reversal. We hypothesized that corticosteroids lower transforming growth factor-beta (TGF-beta) and insulinlike growth factor-I (IGF-I) levels and tissue deposition in wounds and that retinoids stimulate corticosteroid-impaired TGF-beta and ICF-I release and collagen production. Design: Randomized controlled trial. Setting: Wound healing research laboratory. Participants: Animal study. Interventions: Four wire mesh wound cylinders were implanted subcutaneously into the backs of 72 male Sprague-Dawley rats. Wound healing was impaired by a single subcutaneous injection of 6 mg of methylprednisolone acetate (Depo-Medrol). Two preparations of retinoids were used in separate experiments: all-trans-retinoic acid and 9-cis-retinoic acid that were fed orally. Mule Outcome Measures: Hydroxyproline content was measured in the healing tissue and TGF-beta and IGF-I levels were analyzed in the wound fluid. Results: Methylprednisolone treatment significantly decreased TGF-beta and IGF-I levels in the wound fluid and hydroxyproline content in the tissue (P<.05). Oral all-trans- and 9-cis-retinoic acid partially reversed the TGF- and IGF-I decrease and significantly increased hydroxyproline content toward normal levels (P<.05). Oral all-trans-retinoic acid enhanced collagen deposition, TGF- and IGF-I levels over normal chow fed control animals (P<.05). Conclusions: Steroids and retinoids have antagonistic effects on growth factors and collagen deposition in wound healing. These effects can be relevant fur treatment options in a clinical setting. C1 Univ Tubingen, Dept Surg, D-72076 Tubingen, Germany. Univ Calif San Francisco, Dept Surg, San Francisco, CA 94143 USA. Univ Calif Davis, Div Plast Surg, Davis, CA 95616 USA. NCI, Lab Cell Regulat & Carcinogenesis, Bethesda, MD 20892 USA. Davies Med Ctr, Lab Growth & Dev, San Francisco, CA USA. RP Wicke, C (reprint author), Univ Tubingen, Dept Surg, Hoppe Seyler Str 3, D-72076 Tubingen, Germany. NR 35 TC 127 Z9 135 U1 2 U2 10 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0004-0010 J9 ARCH SURG-CHICAGO JI Arch. Surg. PD NOV PY 2000 VL 135 IS 11 BP 1265 EP 1270 DI 10.1001/archsurg.135.11.1265 PG 6 WC Surgery SC Surgery GA 369FN UT WOS:000165055400002 PM 11074878 ER PT J AU Darbeau, MZ Lutz, RJ Collins, WE AF Darbeau, MZ Lutz, RJ Collins, WE TI Simulated lipoprotein transport in the wall of branched arteries SO ASAIO JOURNAL LA English DT Article ID LOW-DENSITY LIPOPROTEINS; BREAKDOWN PRODUCTS; PLATELET-TRANSPORT; RELEASE RATE; ATHEROSCLEROSIS; BLOOD; MODEL; FLOW; LOCALIZATION; JUNCTIONS AB Study of arterial blood flow dynamics improves our understanding of the development of cardiovascular diseases such as atherosclerosis. The transport and accumulation of macromolecules in the arterial wall can be influenced by local fluid mechanics. We used numeric simulations to investigate such transport in a T-junction model. Presumably an in vitro experiment would consist of gel segments inserted in the walls of a mechanical flow T-junction model near branch points where separation and recirculation zones are expected. The transport of low density lipoprotein (LDL) was investigated theoretically at these sites in a two dimensional numeric T-branch model. In the numeric model, the hydraulic conductivity of the porous gel wall segments was varied for a fixed species diffusivity to provide simulations with wall transmural Peclet numbers ranging from 0.3 to 30. Steady state flow patterns in the lumen of the two dimensional T-branch were simulated at Reynolds numbers of 250 and 500, using the software package FIDAP 7.61 to implement the finite element method. The simulations demonstrated that wall Peclet numbers greater than 1.0 were needed to achieve species concentration gradients within the wall that varied in the axial direction, thereby reflecting the influence of disturbed flow and pressure patterns in the lumen. As expected, the transmural concentration gradients were steeper when convection predominated. Blood flow in the lumen can influence the distribution of macromolecules in the arterial wall and needs to be investigated for the relevance to atherosclerosis. C1 Howard Univ, Dept Chem Engn, Washington, DC 20059 USA. NIH, Biomed Engn & Instrumentat Branch, Bethesda, MD 20892 USA. RP Collins, WE (reprint author), Howard Univ, Dept Chem Engn, LK Downing Hall,Room 1011,2300 6th St NW, Washington, DC 20059 USA. NR 26 TC 12 Z9 12 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1058-2916 J9 ASAIO J JI Asaio J. PD NOV-DEC PY 2000 VL 46 IS 6 BP 669 EP 678 DI 10.1097/00002480-200011000-00006 PG 10 WC Engineering, Biomedical; Transplantation SC Engineering; Transplantation GA 374LU UT WOS:000165347300005 PM 11110263 ER PT J AU Phillips, SJ AF Phillips, SJ TI Physiology of wound healing and surgical wound care SO ASAIO JOURNAL LA English DT Article ID MACROPHAGES; ANGIOGENESIS; STIMULATION; INVIVO; GROWTH AB Wound healing is a systemic process, which occurs stepwise and involves the stages of hemostasis, inflammation, and repair. Hemostasis with fibrin formation creates a protective wound scab. The scab provides a surface beneath which cell migration and movement of the wound edges can occur. Inflammation brings nutrients to the area of the wound, removes debris and bacteria, and provides chemical stimuli for wound repair. Repair begins immediately after wounding and proceeds rapidly through the processes of epithelialization, fibroplasia, and capillary proliferation into the healing area. Different tissues have their own normal rates of growth during the process of healing. The optimal rate of healing is approached when factors advantageous to healing are present and factors having the ability to disturb or retard the healing processes are controlled or absent. These factors are discussed. C1 NIH, Natl Lib Med, Bethesda, MD 20894 USA. RP Phillips, SJ (reprint author), NIH, Natl Lib Med, 8600 Rockville Pike, Bethesda, MD 20894 USA. NR 39 TC 15 Z9 15 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1058-2916 J9 ASAIO J JI Asaio J. PD NOV-DEC PY 2000 VL 46 IS 6 BP S2 EP S5 DI 10.1097/00002480-200011000-00029 PG 4 WC Engineering, Biomedical; Transplantation SC Engineering; Transplantation GA 374LU UT WOS:000165347300028 PM 11110286 ER PT J AU Copolov, D Crook, J AF Copolov, D Crook, J TI Biological markers and schizophrenia SO AUSTRALIAN AND NEW ZEALAND JOURNAL OF PSYCHIATRY LA English DT Article DE biological markers; dopamine; homovanillic acid; review; schizophrenia; smooth pursuit eye movements; ventricles ID HOMOVANILLIC-ACID CONCENTRATIONS; POSITRON EMISSION TOMOGRAPHY; D2 DOPAMINE-RECEPTORS; CEREBROSPINAL-FLUID; HEALTHY CONTROLS; OCULOMOTOR ABNORMALITIES; MONOAMINE METABOLITES; HALOPERIDOL CHALLENGE; PROLACTIN RESPONSE; CSF LEVELS AB Objective: The delivery of biological markers for schizophrenia would greatly assist preventative strategies by identifying at-risk individuals who could then be monitored and treated in a manner with a view to minimising subsequent morbidity. This paper aims to present a selection of biological measures that may indicate risk of schizophrenia. Method: A selective and brief review is provided of intensively studied putative markers, including enlarged cerebral ventricles, dopamine D-2 receptor density, amphetamine-stimulated central nervous system dopamine release, plasma homovanillic acid and smooth pursuit eye tracking dysfunction. Results: A number of biological measures have been reported to be correlated with schizophrenia. Conclusions: Presently, none of these measures has satisfactory performance characteristics in terms of predictive validity, noninvasiveness, ease of testing and low cost that would enable their widespread use. However, a few have potential for further investigation and development. C1 Mental Hlth Res Inst Victoria, Parkville, Vic 3052, Australia. NIMH, Clin Brain Disorders Branch, Bethesda, MD 20892 USA. RP Copolov, D (reprint author), Mental Hlth Res Inst Victoria, Locked Bag 11, Parkville, Vic 3052, Australia. NR 47 TC 15 Z9 16 U1 0 U2 5 PU BLACKWELL SCIENCE ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 0004-8674 J9 AUST NZ J PSYCHIAT JI Aust. N. Z. J. Psych. PD NOV PY 2000 VL 34 SU S BP S108 EP S112 DI 10.1080/000486700230 PG 5 WC Psychiatry SC Psychiatry GA 385UX UT WOS:000166023600016 PM 11129294 ER PT J AU Boyce, CA Hoagwood, K Lopez, ML Tarullo, LB AF Boyce, CA Hoagwood, K Lopez, ML Tarullo, LB TI The Head Start Mental Health Research Consortium: New directions for research partnerships SO BEHAVIORAL DISORDERS LA English DT Article AB The Head Start Mental Health Research Consortium (HSMHRC) is a research partnership between the Administration on Children, Youth and Families (ACYF) and the National Institute of Mental Health (NIMH). Funded jointly by these two agencies, a research consortium with five core sites brought together a range of disciplinary perspectives with three primary goals: (1) to develop assessment strategies for identifying young children's social and emotional development, (2) to assess Head Start families' access to mental health services; and (3) to investigate the effectiveness of both universal and targeted intervention models for Head Start children, including those with co-occurring language and behavioral problems. This collaborative mental health research will yield information about the types of mental health needs of children from low-income families, the optimal approaches for assessing these needs, and the impact of research-based intervention models delivered within a Head Start context. C1 NIMH, Bethesda, MD 20852 USA. Adm Children Youth & Families, Commissioners Off Res & Evaluat, Washington, DC USA. RP Boyce, CA (reprint author), NIMH, 6001 Execut Bldg,Room 6200, Bethesda, MD 20852 USA. NR 27 TC 5 Z9 5 U1 0 U2 0 PU COUNCIL CHILDREN BEHAVIORAL DISORDERS PI RESTON PA 1920 ASSOCIATION DR, RESTON, VA 20191-1589 USA SN 0198-7429 J9 BEHAV DISORDERS JI Behav. Disord. PD NOV PY 2000 VL 26 IS 1 BP 7 EP 12 PG 6 WC Psychology, Clinical; Psychology, Educational SC Psychology GA 708ZF UT WOS:000184599700002 ER PT J AU Soltis, J Thomsen, R Matsubayashi, K Takenaka, O AF Soltis, J Thomsen, R Matsubayashi, K Takenaka, O TI Infanticide by resident males and female counter-strategies in wild Japanese macaques (Macaca fuscata) (vol 48, pg 195, 2000) SO BEHAVIORAL ECOLOGY AND SOCIOBIOLOGY LA English DT Correction C1 NIH, Comparat Ethol Lab, Poolesville, MD 20837 USA. RP Soltis, J (reprint author), NIH, Comparat Ethol Lab, Box 529, Poolesville, MD 20837 USA. NR 1 TC 0 Z9 0 U1 0 U2 6 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-5443 J9 BEHAV ECOL SOCIOBIOL JI Behav. Ecol. Sociobiol. PD NOV PY 2000 VL 48 IS 6 BP 490 EP 490 PG 1 WC Behavioral Sciences; Ecology; Zoology SC Behavioral Sciences; Environmental Sciences & Ecology; Zoology GA 376CY UT WOS:000165441700012 ER PT J AU Yang, XN Shears, SB AF Yang, XN Shears, SB TI Multitasking in signal transduction by a promiscuous human Ins(3,4,5,6)P-4 1-kinase/Ins(1,3,4)P-3 5/6-kinase SO BIOCHEMICAL JOURNAL LA English DT Article DE chloride channel; inositol phosphates; signalling ID INOSITOL 1,3,4-TRISPHOSPHATE 5/6-KINASE; RAT-LIVER; 3,4,5,6-TETRAKISPHOSPHATE; KINASE; ENZYME AB We describe a human cDNA encoding 1-kinase activity that inactivates Ins(3,4,5,6)P-4, an inhibitor of chloride-channel conductance that regulates epithelial salt and fluid secretion, as well as membrane excitability. Unexpectedly, we further discovered that this enzyme has alternative positional specificity (5/6-kinase activity) towards a different substrate, namely Ins(1,3,3)P-3. Kinetic data from a recombinant enzyme indicate that Ins(1,3,4)P-3 (K-m = 0.3 muM; V-max = 320 pmol/min per mug) and Ins(3,4,5,6)P-4 (K-m = 0.1 muM; V-max = 780 pmol/min per mug) actively compete for phosphorylation in vivo. This competition empowers the kinase with multitasking capability in several key aspects of inositol phosphate signalling. C1 NIEHS, Inositide Signaling Sect, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Shears, SB (reprint author), NIEHS, Inositide Signaling Sect, Lab Signal Transduct, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 25 TC 49 Z9 52 U1 0 U2 0 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD NOV 1 PY 2000 VL 351 BP 551 EP 555 DI 10.1042/0264-6021:3510551 PN 3 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 372GY UT WOS:000165227000002 PM 11042108 ER PT J AU Li, BB Zhang, HJ Akbar, M Kim, HY AF Li, BB Zhang, HJ Akbar, M Kim, HY TI Negative regulation of cytosolic phospholipase A(2) by melatonin in the rat pineal gland SO BIOCHEMICAL JOURNAL LA English DT Article DE arachidonic acid; gas chromatography/mass spectrometry; isoproterenol; melatonin receptor ID ARACHIDONIC-ACID; N-ACETYLTRANSFERASE; NUCLEAR RECEPTOR; ACTIVATION; PITUITARY; PROTEIN; BIOSYNTHESIS; INHIBITION; MECHANISMS; MODULATION AB In this paper evidence that supports a new role for melatonin as a negative endogenous regulator of cytosolic phospholipase A(2) (cPLA(2)) is presented. When rat pineal glands were incubated in culture, time-dependent release of arachidonic acid (AA) was observed, which was significantly inhibited by a known 85-kDa cPLA(2) inhibitor, methyl arachidonyl fluorophosphonate. Co-incubation with melatonin inhibited the AA release in a concentration-dependent manner, and this decrease was accompanied by a reduction of cPLA(2) protein and mRNA expression. Melatonin-receptor agonists, 2-iodo-N-butanoyl-5-methoxytryptamine and 5-methoxycarbonylamino-N-acetyltryptamine also decreased AA release and cPLA(2) protein and mRNA levels, while pre-incubation with the melatonin receptor antagonists luzindole and 2-phenylmelatonin abolished the melatonin effect. In vivo, as melatonin production reflected a typical diurnal variation, endogenous non-esterified AA and cPLA(2) mRNA levels in the rat pineal gland showed an off-phase diurnal pattern in relation to melatonin levels. Intravenous administration of isoproterenol, which has bean shown to elevate melatonin production, also decreased the levels of non-esterified AA and cPLA(2) mRNA significantly. Direct administration of melatonin to rats by intravenous injection decreased the levels of nonesterified AA, cPLA(2) protein and mRNA in rat pineal glands. In conclusion, melatonin endogenously down-regulates cPLA(2) expression, presumably through melatonin-receptor-mediated processes. C1 NIAAA, Sect Mass Spectrometry, Lab membrane Biochem & Biophys, NIH, Rockville, MD 20852 USA. RP Kim, HY (reprint author), NIAAA, Sect Mass Spectrometry, Lab membrane Biochem & Biophys, NIH, 12425 Parklawn Dr, Rockville, MD 20852 USA. NR 46 TC 10 Z9 11 U1 0 U2 1 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD NOV 1 PY 2000 VL 351 BP 709 EP 716 DI 10.1042/0264-6021:3510709 PN 3 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 372GY UT WOS:000165227000020 PM 11042126 ER PT J AU McKay, RR Szymeczek-Seay, CL Lievremont, JP Bird, GS Zitt, C Jungling, E Luckhoff, A Putney, JW AF McKay, RR Szymeczek-Seay, CL Lievremont, JP Bird, GS Zitt, C Jungling, E Luckhoff, A Putney, JW TI Cloning and expression of the human transient receptor potential 4 (TRP4) gene: localization and functional expression of human TRP4 and TRP3 SO BIOCHEMICAL JOURNAL LA English DT Article DE capacitative calcium entry; calcium channels; HEK-293 cells; patch-clamp; non-selective cation current ID CAPACITATIVE CALCIUM-ENTRY; CA2+-PERMEABLE CATION CHANNEL; OPERATED HTRP3 CHANNELS; PDZ-DOMAIN PROTEIN; CA2+ ENTRY; MOLECULAR-CLONING; PLASMA-MEMBRANE; DROSOPHILA TRP; HUMAN HOMOLOG; SIGNALING COMPLEX AB Mammalian homologues of the Drosophila transient receptor potential (TRP) protein have been proposed to function as ion channels, and in some cases as store-operated or capacitative calcium entry channels. However, for each of the mammalian TRP proteins, different laboratories have reported distinct modes of cellular regulation. In the present study we describe the cloning and functional expression of the human form of TRP4 (hTRP4), and compare its activity with another well studied protein, hTRP3. When hTRP3 was transiently expressed in human embryonic kidney (HEK)-293 cells, basal bivalent cation permeability (barium) was increased. Whole-cell patch-clamp studies of hTRP4 expressed in Chinese hamster ovary cells revealed a constitutively active non-selective cation current which probably underlies the increased bivalent cation entry. Barium entry into hTRP4-transfected HEK-293 cells was not further increased by phospholipase: C (PLC)-linked receptor activation, by intracellular calcium store depletion with thapsigargin, or by a synthetic diacylglycerol, 1-oleoyl-2-acetyl-sn-glycerol (OAG). In contrast, transient expression of hTRP3 resulted in a bivalent cation influx that was markedly increased by PLC-linked receptor activation and by OAG, but not by thapsigargin. Despite the apparent differences in regulation of these two putative channel proteins, green fluorescent protein Fusions of both molecules localized similarly to the plasma-membrane, notably in discrete punctate regions suggestive of specialized signalling complexes. Our findings indicate that while both hTRP4 and hTRP3 can apparently function as cation channels, their putative roles as components of capacitative calcium entry channels are not readily demonstrable by examining their behaviour when exogenously expressed in cells. C1 NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. Rhein Westfal TH Aachen, Inst Physiol, D-52074 Aachen, Germany. RP Putney, JW (reprint author), NIEHS, Lab Signal Transduct, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 48 TC 101 Z9 108 U1 1 U2 2 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD NOV 1 PY 2000 VL 351 BP 735 EP 746 DI 10.1042/0264-6021:3510735 PN 3 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 372GY UT WOS:000165227000023 PM 11042129 ER PT J AU Mimnaugh, EG Yunmbam, MK Li, QD Bonvini, P Hwang, SG Trepel, J Reed, E Neckers, L AF Mimnaugh, EG Yunmbam, MK Li, QD Bonvini, P Hwang, SG Trepel, J Reed, E Neckers, L TI Prevention of cisplatin-DNA adduct repair and potentiation of cisplatin-induced apoptosis in ovarian carcinoma cells by proteasome inhibitors SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE proteasome; ubiquitin; nucleosomal histones; cisplatin; ERCC-1; DNA repair ID NUCLEOTIDE EXCISION-REPAIR; MESSENGER-RNA EXPRESSION; HEAT-SHOCK RESPONSE; 2 HUMAN HOMOLOGS; SACCHAROMYCES-CEREVISIAE; CANCER CELLS; IMMUNOCHEMICAL DETECTION; COMPLEMENTATION GROUP-1; UBIQUITINATED HISTONES; PROTEIN-DEGRADATION AB Histones H2A and H2B are known to be reversibly post-translationally modified by ubiquitination. We previously observed in cultured tumor cells that proteasome inhibition stabilizes polyubiquitinated proteins, depletes unconjugated ubiquitin, and thereby promotes the deubiquitination of nucleosomal histones in chromatin. Provocative indirect evidence suggests that histone ubiquitination/deubiquitination cycles alter chromatin structure, which may limit accessibility of DNA repair proteins to damaged sites. In the present study, we focused on the relationship between the ubiquitination status of histone H2A, the structure of chromatin, and the efficiency of nucleotide excision repair (NER) of cisplatin-DNA adducts in human ovarian carcinoma cells exposed to the antitumor drug cisplatin. Pretreating cells with the proteasome inhibitor lactacystin (LC) or N-acetyl-leucyl-Leucyl-norleucinal (ALLnL) induced deubiquitination of ubiquitinated histone H2A (uH2A) and concomitantly promoted chromatin condensation, increased the extent of cisplatin-DNA adducts, and diminished NER-dependent repair of cisplatin-DNA lesions, compared with control cells treated with cisplatin alone. Both proteasome inhibitors also prevented the increase in ERCC-1 mRNA expression that occurs in cells exposed to cisplatin. Cells treated with the combination of ALLnL and cisplatin underwent apoptosis, as indicated by caspase-dependent poly(ADP-ribose) polymerase (PARP) cleavage, more quickly than cells treated with tither agent alone. Additionally, the combination of ALLnL and cisplatin potently increased p53 levels in cell lysates and stimulated the binding of p53 to chromatin. Together, these observations suggest that proteasome inhibition may be exploited therapeutically for its potential to sensitize ovarian tumor cells to cisplatin. (C) 2000 Elsevier Science Inc. C1 NCI, Tumor Cell Biol Sect, Med Branch, Key W Ctr,Cell & Canc Biol Dept,NIH, Rockville, MD 20850 USA. NCI, Med Ovarian Canc Sect, Med Branch, Dev Therapeut Dept,NIH, Rockville, MD 20850 USA. NCI, Cell Signaling & Oncogenesis Sect, Med Branch, Cell & Canc Biol Dept,NIH, Rockville, MD 20850 USA. RP Mimnaugh, EG (reprint author), NCI, Tumor Cell Biol Sect, Med Branch, Key W Ctr,Cell & Canc Biol Dept,NIH, Suite 300,9610 Med Ctr Dr, Rockville, MD 20850 USA. NR 73 TC 37 Z9 38 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD NOV 1 PY 2000 VL 60 IS 9 BP 1343 EP 1354 DI 10.1016/S0006-2952(00)00455-X PG 12 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 361BY UT WOS:000089702900010 PM 11008128 ER PT J AU Malev, VV Kaulin, YA Bezrukov, SM Gurnev, PA Takemoto, JY Schagina, LV AF Malev, VV Kaulin, YA Bezrukov, SM Gurnev, PA Takemoto, JY Schagina, LV TI Kinetics of opening-closure of syringomycin E channels formed in lipid bilayers SO BIOLOGICHESKIE MEMBRANY LA Russian DT Article ID ION CHANNELS; MEMBRANES; ALAMETHICIN; GRAMICIDIN; MECHANISM; MODELS AB As was shown earlier the cyclic lipodepsipeptide, syringomycin E, when incorporated into planar lipid membranes, forms two types of channels ("small" and "large") that are different in their conductance approximately by a factor of six [Biophys. J.,-1998, 74, 2918-2925]. In this,stydy the dynamics of the SRE-induced transmembrane current at voltage-clamp conditions has been analyzed to clarify mechanisms of these channel formation. As a result, the following conclusions are achieved. The potential-dependent opening/closure of SRE channels in lipid bilayers is accompanied by transitions between three types of clusters including 6-7 SRE molecules and some Lipid molecules. Among such clusters there is the initial one called a predecessor, which is in equilibrium with monomer molecules of SRE at the membrane surface. The other two types of clusters (State 1 and State 2) are formed from the predecessor and their interconversions as well, i.e, the consecutive-parallel mechanism of such transitions take place in the studied system. State 1 turns out to be a non-conducting state and is in equilibrium with small channels determining partially the ionic conductance of lipid bilayers modified by SRE. State 2 corresponds to large SRE channels that provide the main contribution to the conductance of a bilayer, Theoretical consequences resulted from the use of the chemical kinetics concepts are consistent with experimental observations. The established cluster organization of SRE channels, their potential dependent opening/closure and the existence of non-conducting state of the channels are also inherent to some ionic channels of biological membranes. This makes it possible to use SRE channels as a model for natural ionic channels. C1 Russian Acad Sci, Inst Cytol, St Petersburg 194064, Russia. St Petersburg State Univ, Petergof, Russia. Monell Chem Senses Ctr, Philadelphia, PA 19104 USA. NIH, Bethesda, MD 20892 USA. Russian Acad Sci, St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. Utah State Univ, Logan, UT 84322 USA. RP Malev, VV (reprint author), Russian Acad Sci, Inst Cytol, St Petersburg 194064, Russia. RI Takemoto, Jon/A-5309-2011 OI Takemoto, Jon/0000-0001-9919-9168 NR 16 TC 7 Z9 8 U1 0 U2 0 PU MEZHDUNARODNAYA KNIGA PI MOSCOW PA 39 DIMITROVA UL., 113095 MOSCOW, RUSSIA SN 0233-4755 J9 BIOL MEMBRANY JI Biol. Membr. PD NOV-DEC PY 2000 VL 17 IS 6 BP 653 EP 665 PG 13 WC Cell Biology SC Cell Biology GA 389PE UT WOS:000166246300010 ER PT J AU Burroughs, KD Fuchs-Young, R Davis, B Walker, CL AF Burroughs, KD Fuchs-Young, R Davis, B Walker, CL TI Altered hormonal responsiveness of proliferation and apoptosis during myometrial maturation and the development of uterine leiomyomas in the rat SO BIOLOGY OF REPRODUCTION LA English DT Article DE aging; apoptosis; estradiol; female reproductive tract; progesterone; steroid hormones; uterus ID AGING FEMALE RATS; GROWTH-FACTOR-I; ESTROGEN-RECEPTOR; CELL-DEATH; UTERUS; GONADOTROPIN; PROGESTERONE; LOCALIZATION; STIMULATION; ENDOMETRIUM AB Uterine leiomyomas are responsive to the ovarian steroids, estrogen and progesterone; however, a mechanistic understanding of the role of these hormones in the development of this common gynecologic lesion remains to be elucidated. We have used the Eker rat uterine leiomyoma model to investigate how ovarian hormones regulate or promote the growth of these tumors. Proliferative and apoptotic rates were quantitated in normal uterine tissues and leiomyomas in response to endogenous ovarian steroids. In 2- to 4-mo-old animals, cell proliferation in the normal uterus corresponded with high serum levels of steroid hormones during the estrous cycle, and apoptosis occurred in the rat uterus in all cell types following sharp, cyclical declines in serum hormone levels, It is interesting that the responsiveness of uterine mesenchymal cells changed between 4 and 6 mo of age, with significant decreases in both proliferative and apoptotic rates observed in myometrial and stromal cells of cycling animals. Leiomyomas displayed much higher levels of proliferation than did age-matched myometrium; however, their apoptotic index was significantly decreased in comparison with normal myometrium. This disregulation between proliferative and apoptotic responses, which were tightly regulated during ovarian cycling in the normal myometrium, may contribute to the disruption of tissue homeostasis and underlie neoplastic growth of these tumors. C1 Univ Texas, MD Anderson Canc Ctr, Dept Carcinogenesis, Div Sci Pk Res, Smithville, TX 78957 USA. NIEHS, Lab Womens Hlth, Res Triangle Pk, NC 27709 USA. RP Walker, CL (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Carcinogenesis, Div Sci Pk Res, Pk Rd 1C, Smithville, TX 78957 USA. FU NCI NIH HHS [CA16672, CA72253]; NIEHS NIH HHS [ES08263] NR 35 TC 53 Z9 60 U1 0 U2 2 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 NOV PY 2000 VL 63 IS 5 BP 1322 EP 1330 DI 10.1095/biolreprod63.5.1322 PG 9 WC Reproductive Biology SC Reproductive Biology GA 369VT UT WOS:000165089000013 PM 11058535 ER PT J AU Perez, GI Maravei, DV Trbovich, AM Cidlowski, JA Tilly, JL Hughes, FM AF Perez, GI Maravei, DV Trbovich, AM Cidlowski, JA Tilly, JL Hughes, FM TI Identification of potassium-dependent and -independent components of the apoptotic machinery in mouse ovarian germ cells and granulosa cells SO BIOLOGY OF REPRODUCTION LA English DT Article DE apoptosis; follicle; granulosa cells; ovum ID DIEPOXIDE-INDUCED OVOTOXICITY; BCL-2 GENE FAMILY; DNA FRAGMENTATION; ENDONUCLEASE ACTIVITY; FOLLICULAR ATRESIA; RAT GRANULOSA; INTACT-CELLS; IN-VITRO; DEATH; EXPRESSION AB Recent studies with thymocytes have suggested a critical role for intracellular potassium in the regulation of apoptosis. In this study, we examined the pathways of K+ regulation during ovarian cell death. In initial studies, fluorographic analysis demonstrated a significant loss of K+ during apoptosis stimulated by doxorubicin in oocytes and trophic hormone deprivation in granulosa cells. In oocytes, suppression of potassium efflux by potassium-enriched medium prevented condensation, budding, and fragmentation, although it did not block DNA degradation, suggesting the existence of potassium-independent nucleases in oocytes. Culture of granulosa cells in potassium-enriched medium inhibited internucleosomal DNA cleavage, although high-molecular weight DNA cleavage was apparent, suggesting that the nuclease or nucleases responsible for generating 50-kilobase (kb) fragments in these cells is potassium independent. To address this directly, isolated granulosa cell nuclei were stimulated to autodigest their DNA, and internucleosomal, but not large-fragment, cleavage was completely blocked by 150 mM potassium. We next examined whether the proapoptotic caspases are targets for potassium regulation. In cell-free assays, processing of pro-interleukin-1 beta and proteolysis of cellular actin by recombinant caspase-1 and caspase-3, respectively, were suppressed by the presence of 150 mM potassium. Other monovalent ions (NaCl, LiCl) exerted a similar effect in these cell-free assays. Thus, in oocytes and granulosa cells, potassium efflux appears to occur early in the cell death program and may regulate a number of apoptotic events including caspase activity and internucleosomal DNA cleavage. However, there also exist novel potassium-independent pathways in both ovarian germ cells and somatic cells that signal certain apoptotic events, such as large-fragment DNA cleavage. C1 Univ N Carolina, Dept Biol, Charlotte, NC 28223 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Obstet & Gynecol,Vincent Ctr Reprod Biol, Boston, MA 02114 USA. NIEHS, Lab Integrat Biol, NIH, Res Triangle Pk, NC 27709 USA. RP Hughes, FM (reprint author), Univ N Carolina, Dept Biol, 9201 Univ City Blvd, Charlotte, NC 28223 USA. FU NIA NIH HHS [R01-AG12279]; NICHD NIH HHS [R01-HD34226]; NIEHS NIH HHS [R01-ES08430] NR 50 TC 43 Z9 48 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 NOV PY 2000 VL 63 IS 5 BP 1358 EP 1369 DI 10.1095/biolreprod63.5.1358 PG 12 WC Reproductive Biology SC Reproductive Biology GA 369VT UT WOS:000165089000017 PM 11058539 ER PT J AU Ye, CI Xiao, RP Izu, LT Cheng, HP Kuschel, M Spurgeon, H Lakatta, EG AF Ye, CI Xiao, RP Izu, LT Cheng, HP Kuschel, M Spurgeon, H Lakatta, EG TI G(i)-dependent localization of beta(2)-adrenergic receptor signaling to L-type Ca(2+)channels SO BIOPHYSICAL JOURNAL LA English DT Article ID BETA-ADRENERGIC STIMULATION; DEPENDENT PROTEIN-KINASE; CALCIUM CHANNELS; CARDIAC MYOCYTES; CA2+ CHANNELS; HEART-CELLS; VENTRICULAR MYOCYTES; ANCHORING PROTEIN; ADENYLYL CYCLASE; PERTUSSIS-TOXIN AB A plausible determinant of the specificity of receptor signaling is the cellular compartment over which the signal is broadcast. In rat heart, stimulation of beta (1)-adrenergic receptor (beta (1)-AR), coupled to G(s)-protein, or beta (2)-AR, coupled to G(s)- and G(i)-proteins, both increase L-type Ca2+ current, causing enhanced contractile strength. But only beta (1)-AR stimulation increases the phosphorylation of phospholamban, troponin-l, and C-protein, causing accelerated muscle relaxation and reduced myofilament sensitivity to Ca2+. beta (2)-AR stimulation does not affect any of these intracellular proteins. We hypothesized that beta (2)-AR signaling might be localized to the cell membrane. Thus we examined the spatial range and characteristics of beta (1)-AR and beta (2)-AR signaling on their common effector, L-type Ca2+ channels. Using the cell-attached patch-clamp technique, we show that stimulation of beta (1)-AR or beta (2)-AR in the patch membrane, by adding agonist into patch pipette, both activated the channels in the patch. But when the agonist was applied to the membrane outside the patch pipette, only beta (1)-AR stimulation activated the channels. Thus, beta (1)-AR signaling to the channels is diffusive through cytosol, whereas beta (2)-AR signaling is localized to the cell membrane. Furthermore, activation of Gi is essential to the localization of beta (2)-AR signaling because in pertussis toxin-treated cells, beta (2)-AR signaling becomes diffusive. Our results suggest that the dual coupling of beta (2)-AR to both G(s)- and G(i)-proteins leads to a highly localized beta (2)-AR signaling pathway to modulate sarcolemmal L-type Ca2+ channels in rat ventricular myocytes. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. Univ Maryland, Sch Med, Dept Physiol Cardiol, Baltimore, MD 21201 USA. RP Xiao, RP (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Izu, Leighton/C-4066-2015 NR 44 TC 82 Z9 86 U1 0 U2 7 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD NOV PY 2000 VL 79 IS 5 BP 2547 EP 2556 PG 10 WC Biophysics SC Biophysics GA 370CE UT WOS:000165104500027 ER PT J AU Polozova, A Litman, BJ AF Polozova, A Litman, BJ TI Cholesterol dependent recruitment of di22 : 6-PC by a G protein-coupled receptor into lateral domains SO BIOPHYSICAL JOURNAL LA English DT Article ID DIFFERENTIAL SCANNING CALORIMETRY; ELECTRON-SPIN-RESONANCE; SEGMENT DISK MEMBRANES; LIPID DOMAINS; DOCOSAHEXAENOIC ACID; BOVINE RHODOPSIN; BIOLOGICAL-MEMBRANES; ROTATIONAL DIFFUSION; RETINAL CHROMOPHORE; MOLECULAR PROBES AB Bovine rhodopsin was reconstituted into mixtures of didocosahexaenoylphosphatidylcholine (di22:6-PC), dipalmitoylphosphatidyIcholine (di16:O-PC), sn-1-palmitoyl-sn-2-docosahexaenoylphosphatidylcholine (16:0, 22:6-PC) and cholesterol. Rhodopsin denaturation was examined by using high-sensitivity differential scanning calorimetry. The unfolding temperature was increased at lower levels of lipid unsaturation, but the highest temperature was detected for native disk membranes: di22:6-PC < 16:0,22:6-PC < di16:0,18:1-PC < native disks. The incorporation of 30 mol% of cholesterol resulted in 2-4C increase of denaturation temperature in all reconstituted systems examined. From the analysis of van't Hoff's and calorimetric enthalpies, it was concluded that the presence of cholesterol in di22:6-PC-containing bilayers induces a level of cooperativity in rhodopsin unfolding. Fluorescence resonance energy transfer (FRET), using lipids labeled at the headgroup with pyrene (Py) as donors and rhodopsin retinal group as acceptor of fluorescence, was used to study rhodopsin association with lipids. Higher FRET efficiencies detected for di22:6-PE-Py, compared to di16:O-PE-Py, in mixed di22:6-PC-di16:O-PC-cholesterol bilayers, indicate preferential segregation of rhodopsin with polyunsaturated lipids. The effective range of the rhodopsin-lipid interactions facilitating cluster formation exceeds two adjacent lipid layers. In similar mixed bilayers containing no cholesterol, cluster formation was absent at temperatures above lipid phase transition, indicating a crucial role of cholesterol in microdomain formation. C1 NIAAA, Lab Membrane Biochem & Biophys, NIH, Rockville, MD 20852 USA. RP Litman, BJ (reprint author), NIAAA, Lab Membrane Biochem & Biophys, NIH, 12420 Parklawn Dr, Rockville, MD 20852 USA. RI Polozova, Alla/C-2395-2008 NR 54 TC 76 Z9 76 U1 2 U2 6 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD NOV PY 2000 VL 79 IS 5 BP 2632 EP 2643 PG 12 WC Biophysics SC Biophysics GA 370CE UT WOS:000165104500034 PM 11053136 ER PT J AU Ma, BY Tsai, CJ Nussinov, R AF Ma, BY Tsai, CJ Nussinov, R TI A systematic study of the vibrational free energies of polypeptides in folded and random states SO BIOPHYSICAL JOURNAL LA English DT Article ID COLD-DENATURATION; GLOBULAR-PROTEINS; BINDING; PEPTIDES; ENTROPY; THERMODYNAMICS; SIMULATIONS; DYNAMICS; LANDSCAPES; UBIQUITIN AB Molecular vibrations, especially low frequency motions, may be used as an indication of the rigidity or the flatness of the protein folding energy landscape. We have studied the vibrational properties of native folded as well as random coil structures of more than 60 polypeptides. The picture we obtain allows us to perceive how and why the energy landscape progressively rigidifies while still allowing potential flexibility. Compared with random coil structures, both oc-helices and beta -hairpins are vibrationally more flexible. The vibrational properties of loop structures are similar to those of the corresponding random coil structures. Inclusion of an alpha -helix tends to rigidify peptides and so-called building blocks of the structure, whereas the addition of a beta -structure has less effect. When small building blocks coalesce to form larger domains, the protein rigidifies. However, some folded native conformations are still found to be vibrationally more flexible than random coil structures, for example, beta (2)-microglobulin and the SH3 domain. Vibrational free energy contributes significantly to the thermodynamics of protein folding and affects the distribution of the conformational substates. We found a weak correlation between the vibrational folding energy and the protein size, consistent with both previous experimental estimates and theoretical partition of the heat capacity change in protein folding. C1 NCI, Frederick Canc Res & Dev Ctr, Intramural Res Support Program, SAIC,Intramural Res Support Program, Frederick, MD 21702 USA. Tel Aviv Univ, Sackler Fac Med, Sackler Inst Mol Med, Dept Human Genet & Mol Med, IL-69978 Tel Aviv, Israel. RP Nussinov, R (reprint author), NCI, Frederick Canc Res Facil, Bldg 469,Room 151, Frederick, MD 21702 USA. RI Ma, Buyong/F-9491-2011 OI Ma, Buyong/0000-0002-7383-719X FU NCI NIH HHS [N01-CO-56000] NR 38 TC 16 Z9 16 U1 0 U2 1 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD NOV PY 2000 VL 79 IS 5 BP 2739 EP 2753 PG 15 WC Biophysics SC Biophysics GA 370CE UT WOS:000165104500045 PM 11053147 ER PT J AU Huppi, K Henderson, D Siwarski, D Hochman, J Bergel, M Tuchscherer, G AF Huppi, K Henderson, D Siwarski, D Hochman, J Bergel, M Tuchscherer, G TI Generation of monospecific peptide antibodies to the DNA binding domain of p53 SO BIOTECHNIQUES LA English DT Article ID BREAST-CANCER PATIENTS; CELLULAR PROTEIN P53; MUTATIONS; LYMPHOMA; TUMORS AB The DNA binding domain (DBD) is the most mutated region of p53 in tumors and has proven to be relatively resistant to the generation of specific antibodies. Template assembled synthetic peptide (TASP) synthesis of a peptide derived from the DBD creates a highly immunogenic molecule without the need for large carriers such as keyhole limpet hemocyanin (KLH). In addition, a rapid means of generating monoclonal antibodies can be achieved through immunization in conjuction with ABL/ MYC retrovirus injection into recipient mice. In this paper, we demonstrate that an antibody generated by this means, KH2, reacts specifically with the DBD of p53. To date, this is the first example of a peptide immunogen used successfully in ABL/MYC monoclonal antibody production. KHZ is also the first example of a monospecific antibody that directly binds to and, by definition, assumes the conformation of the DNA binding region of p53. C1 NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. Hebrew Univ Jerusalem, Jerusalem, Israel. Univ Lausanne, Lausanne, Switzerland. RP Huppi, K (reprint author), NCI, Genet Lab, NIH, Bldg 37,Room 2B-21, Bethesda, MD 20892 USA. NR 24 TC 0 Z9 0 U1 0 U2 2 PU EATON PUBLISHING CO PI NATICK PA 154 E. CENTRAL ST, NATICK, MA 01760 USA SN 0736-6205 J9 BIOTECHNIQUES JI Biotechniques PD NOV PY 2000 VL 29 IS 5 BP 1100 EP + PG 6 WC Biochemical Research Methods; Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 373EF UT WOS:000165274900024 PM 11084873 ER PT J AU Flinn, IW Kopecky, KJ Foucar, MK Head, D Bennett, JM Huchison, R Corbett, W Cassileth, P Habermann, T Golomb, H Rai, K Eisenhauer, E Appelbaum, F Cheson, B Grever, MR AF Flinn, IW Kopecky, KJ Foucar, MK Head, D Bennett, JM Huchison, R Corbett, W Cassileth, P Habermann, T Golomb, H Rai, K Eisenhauer, E Appelbaum, F Cheson, B Grever, MR TI Long-term follow-up of remission duration, mortality, and second malignancies in hairy cell leukemia patients treated with pentostatin SO BLOOD LA English DT Article ID NATIONAL-CANCER-INSTITUTE; 2-CHLORODEOXYADENOSINE; 2'-DEOXYCOFORMYCIN; INTERFERON; INDUCTION; THERAPY AB The nucleoside analogue, pentostatin, has demonstrated high complete response rates and long relapse-free survival times in patients with hairy cell leukemia, a disease that historically had been unresponsive to treatment. Long-term data on duration of overall survival and relapse-free survival and incidence of subsequent malignancies with this agent are lacking. Patients completing the treatment phase of a randomized, intergroup study who received pentostatin as an initial treatment or who crossed over after failure of interferon alpha were followed for survival, relapse, and diagnosis of subsequent malignancies. Two hundred forty-one patients treated with pentostatin as initial therapy (n = 154) or who crossed over after failure of interferon alpha(n = 87) were followed for a median duration of 9.3 years. Estimated 5- and 10-year survival rates (95% confidence interval) for all patients combined were 90% (87%-94%) and 81% (75%-86%), respectively. In the 173 patients with a confirmed complete response to pentostatin treatment, 5- and 10-year relapse-free survival rates were 85% (80%-91%) and 67% (58%-76%), respectively. Survival curves for patients initially treated with pentostatin and those crossed over were similar. Only 2 of 40 deaths were attributed to hairy cell leukemia. The mortality rate and incidence of subsequent malignancies were not higher than expected in the general population. Pentostatin is a highly effective regimen for hairy cell leukemia that produces durable complete responses. Subsequent malignancies do not appear to be increased with pentostatin treatment. (C) 2000 by The American Society of Hematology. C1 Johns Hopkins Univ, Baltimore, MD USA. SW Oncol Grp, Ctr Stat, Seattle, WA USA. Univ New Mexico Hosp, Albuquerque, NM USA. St Jude Childrens Res Hosp, Memphis, TN 38105 USA. Univ Rochester, Rochester, NY USA. SUNY Hlth Sci Ctr, Syracuse, NY 13210 USA. Kingston Gen Hosp, Kingston, ON K7L 2V7, Canada. Univ Miami, Miami, FL 33152 USA. Mayo Clin, Rochester, MN USA. Univ Chicago, Chicago, IL 60637 USA. Long Isl Jewish Hosp, New Hyde Park, NY USA. Puget Sound Oncol Consortium, Seattle, WA USA. Natl Canc Inst Canada, Clin Tirals Grp, Kingston, ON, Canada. NCI, Bethesda, MD 20892 USA. Ohio State Univ, Columbus, OH 43210 USA. RP Flinn, IW (reprint author), Operat Off, SW Oncol Grp SWOG8691, 14980 Omicron Dr, San Antonio, TX 78245 USA. FU NCI NIH HHS [CA04920, CA20319, CA38926] NR 26 TC 98 Z9 100 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 NOV 1 PY 2000 VL 96 IS 9 BP 2981 EP 2986 PG 6 WC Hematology SC Hematology GA 367RA UT WOS:000090074200008 PM 11049974 ER PT J AU Sandbrink, F Syed, NA Fujii, MD Dalakas, MC Floeter, MK AF Sandbrink, F Syed, NA Fujii, MD Dalakas, MC Floeter, MK TI Motor cortex excitability in stiff-person syndrome SO BRAIN LA English DT Article DE stiff-person syndrome; transcranial magnetic stimulation; intracortical excitation; intracortical inhibition; silent period ID TRANSCRANIAL MAGNETIC STIMULATION; SILENT PERIOD; CORTICAL EXCITABILITY; CORTICOCORTICAL INHIBITION; SYNAPTIC ORGANIZATION; NEURON EXCITABILITY; EVOKED-RESPONSES; HUMANS; BRAIN; AUTOANTIBODIES AB Muscle stiffness in stiff-person syndrome (SPS) is produced by continuous, involuntary firing of motor units that is thought to be caused by an autoimmune mediated dysfunction of GABA-ergic inhibitory neurones. We have postulated that the loss of GABA-ergic inputs from spinal interneurones alone is insufficient to produce tonic firing of motor neurones and that excessive supraspinal excitation could also play a role. To determine whether SPS is associated with dysfunction in supraspinal GABA-ergic neurones, we assessed the excitability of the motor cortex with transcranial magnetic stimulation (TMS) in seven SPS: patients and seven age-matched healthy volunteers, SPS patients had normal central motor conduction times, normal thresholds for motor evoked potentials (MEPs) in leg muscles, and a normal MEP stimulus versus response recruitment curve with increasing TMS intensities in resting hand and leg muscles. Cortical silent periods were shortened in leg muscles. Intracortical inhibition and excitation were assessed while recording from the abductor pollicis brevis, using a paired pulse TMS paradigm with subthreshold conditioning stimuli, Patients had decreased inhibition and markedly increased facilitation at short intervals. Using paired suprathreshold TMS, patients exhibited increased facilitation at 20- and 40-ms intervals. These results point to a hyperexcitability of the motor cortex in SPS, which could be explained by impairment of supraspinal GABA-ergic neurones, leading to an impaired balance between inhibitory and excitatory intracortical circuitry. C1 NINDS, Electromyog Sect, NIH, Bethesda, MD 20892 USA. NINDS, Neuromuscular Dis Sect, NIH, Bethesda, MD 20892 USA. RP Floeter, MK (reprint author), 10 Ctr Dr,MSC 1404, Bethesda, MD 20892 USA. NR 37 TC 37 Z9 38 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0006-8950 J9 BRAIN JI Brain PD NOV PY 2000 VL 123 BP 2231 EP 2239 DI 10.1093/brain/123.11.2231 PN 11 PG 9 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 375QR UT WOS:000165413200005 PM 11050023 ER PT J AU Clark, EC Baxter, LR Dure, LS Ackermann, RF Kemp, GF Bachus, SE AF Clark, EC Baxter, LR Dure, LS Ackermann, RF Kemp, GF Bachus, SE TI Mammal-like striatal functions in Anolis II. Distribution of dopamine D-1 and D-2 receptors, and a laminar pattern of basal ganglia sub-systems SO BRAIN BEHAVIOR AND EVOLUTION LA English DT Article DE basal ganglia; dopamine; enkephalin; Anolis ID OBSESSIVE-COMPULSIVE DISORDER; SUBTHALAMIC NUCLEUS; MESSENGER-RNA; RAT STRIATUM; D2 DOPAMINE; ACETYLCHOLINE-RELEASE; GLOBUS-PALLIDUS; SPINY NEURONS; SUBSTANCE-P; BRAIN AB We used in situ autoradiographic ligand binding methods to determine the occurrence and distribution of dopamine D-1 and D-2 receptor sub-types in the anole lizard, Anolis carolinensis, Both were present and exhibited pharmacological specificity characteristics similar to those described for mammals. However, unlike in mammals where in the neostriatum [outside the nucleus accumbens/olfactory tubercle complex (NA/OT)] these receptors exhibit only slight dorsolateral (D-2 high, D-1 low) to ventromedial (D-1 high, D-2 low) gradients that co-mingle extensively, in the anole striatum outside the NA/OT there was a striking laminar pattern, with little if any overlap between D-2 (high in a dorsal band) and D-1 thigh ventral to the D-2 band) distributions. As D-1 receptors are related to the direct and D-2 to the indirect basal ganglia (BG) subsystems in mammals, we also determined anole striatal distributions of pre-proenkephalin mRNA, a marker for striatal efferents to the indirect BG subsystem in mammals, Here, too, there was a striking laminar pattern, with pre-proenkephalin mRNA in a band similar to that seen for Dg receptors. The crisp neuroanatomical separation between these classic BG subsystem markers in Anolis striatum make this species attractive for the study of such systems' functions during behavior. Copyright (C) 2001 S. Karger AG, Basel. C1 NIMH, Clin Brain Disorders Branch, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Sch Med, Crump Inst Biomed Imaging, Los Angeles, CA USA. Univ Calif Los Angeles, Sch Med, DoE,Dept Psychiat, Lab Struct Biol & Mol Med,Lab Nucl Med, Los Angeles, CA USA. Univ Calif Los Angeles, Sch Med, DoE,Dept Pharmacol, Lab Struct Biol & Mol Med,Lab Nucl Med, Los Angeles, CA USA. Univ Alabama, Dept Neurol, Birmingham, AL 35294 USA. Univ Alabama, Dept Pediat, Birmingham, AL 35294 USA. Univ Alabama, Dept Neurobiol, Birmingham, AL 35294 USA. Univ Alabama, Dept Psychol, Birmingham, AL 35294 USA. Univ Alabama, Dept Psychiat & Behav Neurobiol, Birmingham, AL 35294 USA. RP Baxter, LR (reprint author), Sparks Res Ctr 1001, 1720 7th Ave, Birmingham, AL 35294 USA. RI Dure, Leon/B-3243-2008 NR 52 TC 13 Z9 13 U1 0 U2 0 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 0006-8977 J9 BRAIN BEHAV EVOLUT JI Brain Behav. Evol. PD NOV PY 2000 VL 56 IS 5 BP 249 EP 258 DI 10.1159/000047208 PG 10 WC Behavioral Sciences; Neurosciences; Zoology SC Behavioral Sciences; Neurosciences & Neurology; Zoology GA 406YW UT WOS:000167244600002 PM 11251317 ER PT J AU Keane, MM Rubinstein, Y Cuello, M Ettenberg, SA Banerjee, P Nau, MM Lipkowitz, S AF Keane, MM Rubinstein, Y Cuello, M Ettenberg, SA Banerjee, P Nau, MM Lipkowitz, S TI Inhibition of NF-kappa B activity enhances TRAIL mediated apoptosis in breast cancer cell lines SO BREAST CANCER RESEARCH AND TREATMENT LA English DT Article DE apoptosis; breast cancer; caspases; NF-kappa B; TRAIL ID FADD-DEPENDENT APOPTOSIS; DEATH DOMAIN; LIGAND TRAIL; DECOY RECEPTORS; ACTIVATION; FAMILY; EXPRESSION; CASPASE-8; INDUCTION; MELANOMA AB Most breast cancer cell lines are resistant to TNF-related apoptosis inducing ligand (TRAIL) induced apoptosis. In sensitive breast cancer cell lines TRAIL rapidly induces the cleavage and activation of caspases leading to the subsequent cleavage of downstream caspase substrates. In contrast, there is no caspase activation in the resistant cell lines. The transcription factor NF-kappaB can inhibit apoptosis induced by a variety of stimuli including activation of death receptors. We investigated whether NF-kappaB contributes to the resistance of breast cancer cells to TRAIL induced apoptosis. All of the resistant breast cancer cell lines expressed NF-kappaB and had detectable NF-kappaB activity in nuclear extracts prior to treatment with TRAIL. Upon TRAIL treatment, a significant increase in NF-kappaB activity was seen in most of the cell lines. To directly test if NF-kappaB activity contributes to the resistance of these cell lines to TRAIL, we transiently transfected the resistant cell lines with an inhibitor of NF-kappaB (I kappaB DeltaN) and measured TRAIL induced apoptosis in control and transfected cells. All of the resistant cell lines tested showed an increase in TRAIL induced apoptosis when transfected with the I kappaB DeltaN. These results demonstrate that TRAIL resistant breast cancer cells fail to rapidly activate the apoptotic machinery but they do activate NF-kappaB. Inhibition of NF-kappaB activity increases the sensitivity to TRAIL mediated apoptosis in resistant cells. These results suggest that agents which inhibit NF-kappaB should increase the clinical efficacy of TRAIL in breast cancer cells. C1 NCI, Dept Genet, Med Branch, Natl Naval Med Ctr, Bethesda, MD 20889 USA. Uniformed Serv Univ Hlth Sci, Cell & Mol Biol Program, Bethesda, MD 20814 USA. RP Lipkowitz, S (reprint author), NCI, Dept Genet, Med Branch, Natl Naval Med Ctr, Blg 8,Rm 5101, Bethesda, MD 20889 USA. NR 42 TC 78 Z9 82 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0167-6806 J9 BREAST CANCER RES TR JI Breast Cancer Res. Treat. PD NOV PY 2000 VL 64 IS 2 BP 211 EP 219 DI 10.1023/A:1006458407515 PG 9 WC Oncology SC Oncology GA 375AT UT WOS:000165377900007 PM 11194457 ER PT J AU Duffey, DC Crowl-Bancroft, CV Chen, Z Ondrey, FG Nejad-Sattari, M Dong, G Van Waes, C AF Duffey, DC Crowl-Bancroft, CV Chen, Z Ondrey, FG Nejad-Sattari, M Dong, G Van Waes, C TI Inhibition of transcription factor nuclear factor-kappa B by a mutant inhibitor-kappa B alpha attenuates resistance of human head and neck squamous cell carcinoma to TNF-alpha caspase-mediated cell death SO BRITISH JOURNAL OF CANCER LA English DT Article DE transcription factors; squamous cell carcinoma; NF-kappa B; cytokins; TNF-alpha ID TUMOR-NECROSIS-FACTOR; PROINFLAMMATORY CYTOKINE EXPRESSION; INDUCED APOPTOSIS; INTERFERON-GAMMA; C-MYC; CONSTITUTIVE ACTIVATION; DNA FRAGMENTATION; FACTOR RECEPTORS; CYCLE; GROWTH AB Tumour necrosis factor-alpha (TNF-(alpha) is a cytokine that can induce cell death of different cancers via a cellular cascade of proteases, the caspases. However, TNF-alpha has been detected in tumour and serum of patients with head and neck squamous cell carcinoma (HNSCC), and tumour cell lines derived from this environment often exhibit resistance to TNF-alpha -induced cell death. Cell death mediated by TNF-alpha and caspases may be inhibited by cytoprotective genes regulated by transcription factor nuclear factor-kappaB (NF-kappaB). We recently showed that NF kappaB is constitutively activated in HNSCC, and that inhibition of NF-kappaB by expression of a nondegradable mutant inhibitor of NF-kappaB, I kappa -B alphaM, markedly decreased survival and growth of HNSCC cells in vivo. In the present study, we examined the TNF-alpha sensitivity and response of HNSCC with constitutively active NF-kappaB, and of HNSCC cells in which NF-kappaB is inhibited by stable expression of a dominant negative mutant inhibitor, I kappaB alphaM. Human lines UM-SCC-9, 11B and 38, previously shown to exhibit constitutive activation of NF-kappaB, were found to be highly resistant to growth inhibition by up to 10(4) U/ml of TNF-alpha in 5 day MTT assay. These TNF-a resistant HNSCC lines expressed TNF receptor I, and exhibited constitutive and TNF-a-inducible activation of NF-kappaB as demonstrated by nuclear localization of NF-kappaB p65 by immunohistochemistry. UM-SCC-9 I11 cells which stably expressed an inhibitor of NF-kappaB, I kappaB alpham, were susceptible to TNF-alpha -induced growth inhibition. DNA cell cycle analysis revealed that TNF-alpha induced growth inhibition was associated with accumulation of cells with sub-G0/G1 DNA content. Cell death was demonstrated by trypan blue staining, and was blocked by caspase Inhibitor. We conclude that HNSCC that exhibit constitutive and TNF-alpha -inducible activation of transcription factor NF-kappaB are resistant to TNF-alpha, and that inhibition of NF-kappaB sensitizes HNSCC to TNF-alpha caspase-mediated cytotoxicity. The demonstration of the role of activation of NF-kappaB in resistance of HNSCC to TNF-alpha may be helpful in the identification of potential targets for pharmacological, molecular and immune therapy of HNSCC. (C) 2000 Cancer Research Campaign. C1 Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. Howard Hughes Med Inst, Bethesda, MD 20817 USA. RP Van Waes, C (reprint author), Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. FU NIDCD NIH HHS [ZO1-DC-00016] NR 64 TC 45 Z9 49 U1 1 U2 6 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD NOV PY 2000 VL 83 IS 10 BP 1367 EP 1374 DI 10.1054/bjoc.2000.1423 PG 8 WC Oncology SC Oncology GA 372BM UT WOS:000165213800017 PM 11044363 ER PT J AU Samuels, J Nestadt, G Bienvenu, OJ Costa, PT Riddle, MA Liang, KY Hoehn-Saric, R Grados, MA Cullen, BAM AF Samuels, J Nestadt, G Bienvenu, OJ Costa, PT Riddle, MA Liang, KY Hoehn-Saric, R Grados, MA Cullen, BAM TI Personality disorders and normal personality dimensions in obsessive-compulsive disorder SO BRITISH JOURNAL OF PSYCHIATRY LA English DT Article ID AXIS-II; ANXIETY; TEMPERAMENT; COMORBIDITY; CHARACTER; MODEL AB Background Little is known about personality disorders and normal personality dimensions in relatives of patients with obsessive-compulsive disorder (OCD). Aims To determine whether specific personality characteristics are part of a familiar spectrum of OCD. Method Clinicians evaluated personality disorders in 72 OCD case and 72 control probands and 198 case and 297 control first-degree relatives. The self completed Revised NEO Personality Inventory was used for assessment of normal personality dimensions. The prevalence of personality disorders and scores on normal personality dimensions were compared between case and control probands and between case and control relatives. Results Case probands and case relatives had a high prevalence of obsessive-compulsive personality disorder (OCPD) and high neuroticism scores. Neuroticism was associated with OCPD in case but not control relatives. Conclusions Neuroticism and OCPD may share a common familiar aetiology with OCD. Declaration of interest Supported by National Institutes of Health grants R0I MH 50214 and NIH/NCRR/OPD-GCRC RR00052. C1 Johns Hopkins Univ, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21205 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. Johns Hopkins Univ, Sch Med, Dept Pediat, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Publ Hlth, Dept Biostat, Baltimore, MD 21205 USA. RP Samuels, J (reprint author), Johns Hopkins Hosp, Dept Psychiat & Behav Sci, Meyer 4-181,600 N Wolfe St, Baltimore, MD 21287 USA. RI Liang, Kung-Yee/F-8299-2011; OI Samuels, Jack/0000-0002-6715-7905; Costa, Paul/0000-0003-4375-1712 FU NCRR NIH HHS [RR00052]; NIMH NIH HHS [R01MH50214] NR 29 TC 131 Z9 137 U1 5 U2 17 PU ROYAL COLLEGE OF PSYCHIATRISTS PI LONDON PA BRITISH JOURNAL OF PSYCHIATRY 17 BELGRAVE SQUARE, LONDON SW1X 8PG, ENGLAND SN 0007-1250 J9 BRIT J PSYCHIAT JI Br. J. Psychiatry PD NOV PY 2000 VL 177 BP 457 EP 462 DI 10.1192/bjp.177.5.457 PG 6 WC Psychiatry SC Psychiatry GA 372TK UT WOS:000165250000014 PM 11060001 ER PT J AU Clughtai, T Chen, LQ Salasidis, G Nguyen, D Tchervenkov, C Morin, JF AF Clughtai, T Chen, LQ Salasidis, G Nguyen, D Tchervenkov, C Morin, JF TI Clips versus suture technique: Is there a difference? SO CANADIAN JOURNAL OF CARDIOLOGY LA English DT Article DE bypass; coronary disease; cost-benefit analysis ID SKIN CLOSURE; OPERATIONS; TRIAL AB INTRODUCTION: Coronary artery bypass grafting (CABG) is one of the most common procedures performed today, and wound complications are a major source of morbidity and cost. OBJECTIVE: To determine whether there is any difference in wound outcome (including cost in a Canadian context) between a subcuticular suture technique and skin stapling technique for closure of sternal and leg incisions in CABG patients. PATIENTS AND METHODS: One hundred and sixty-two patients undergoing CABG were prospectively, randomly placed to have their sternal and leg incisions closed with either a subcuticular suture technique or with a skin clip. Data were obtained through chart review, in-hospital assessments and follow-up visits. Nonblinded assessments were made regarding wound leakage, inflammation, infection, necrosis, swelling, dehiscence and cosmesis. Each of the parameters was graded on a scale from 1 to 4. The cost was evaluated in Canadian dollars. RESULTS: There were trends toward increased rates of in-hospital sternal (P=0.09) and leg (P=0.11) incision inflammation when the wounds were closed with skin clips. There was a significantly greater (P=0.05) rate of sternal wound infection with clips, as well as a tendency (P=0.15) toward a greater rate of mediastinitis at follow-up assessment. Cosmetic outcome was similar for both groups. The cost incurred was significantly greater when skin clips were used for closure. There was a greater than threefold difference, which translates to a greater than $10,000 difference over one year. CONCLUSIONS: Closure with a subcuticular technique achieves better outcomes than the use of skin clips. When factoring in the increased cost incurred by using clips, as well as other intangible factors such as surgical skill acquisition, subcuticular suture closure appears to be a favourable method of wound closure in CABG patients compared with the use of skin stapling techniques. C1 McGill Univ, Montreal Gen Hosp, Ctr Hlth, Dept Cardiothorac Surg, Montreal, PQ H3G 1A4, Canada. Hop Hotel Dieu, Montreal, PQ H2W 1T8, Canada. St Michaels Hosp, Toronto, ON M5B 1W8, Canada. NIH, Bethesda, MD 20892 USA. Univ Montreal, Ctr Hlth, Montreal, PQ, Canada. McGill Univ, Ctr Hlth, Montreal, PQ, Canada. RP Morin, JF (reprint author), McGill Univ, Montreal Gen Hosp, Ctr Hlth, Dept Cardiothorac Surg, L9 513, Montreal, PQ H3G 1A4, Canada. NR 11 TC 0 Z9 0 U1 0 U2 1 PU PULSUS GROUP INC PI OAKVILLE PA 2902 S SHERIDAN WAY, OAKVILLE, ONTARIO L6J 7L6, CANADA SN 0828-282X J9 CAN J CARDIOL JI Can. J. Cardiol. PD NOV PY 2000 VL 16 IS 11 BP 1403 EP 1407 PG 5 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 382BV UT WOS:000165799300006 ER PT J AU Brinton, LA Lubin, JH Burich, MC Colton, T Brown, SL Hoover, RN AF Brinton, LA Lubin, JH Burich, MC Colton, T Brown, SL Hoover, RN TI Breast cancer following augmentation mammoplasty (United States) SO CANCER CAUSES & CONTROL LA English DT Article DE breast cancer; breast implants; incidence; mortality; prognosis; silicone ID IMPLANTS; WOMEN; RISK; DIAGNOSIS; CARCINOMA; SURVIVAL; SILICONE; MAMMAPLASTY; MASS AB Objective: Although clinical reports have raised concern that breast implants may either increase the risk of breast cancer or delay its diagnosis, epidemiologic studies have generally shown implant recipients to be at a reduced risk of subsequent breast cancer. A large retrospective cohort study was undertaken to clarify effects of cosmetic breast implantation. Methods: Medical records of 13,488 women receiving cosmetic implants at 18 plastic surgery practices and a group of 3936 patients who received other types of plastic surgery at the same practices were reviewed and information abstracted. Questionnaires were sent to all subjects located as alive, with 71% being completed. Attempts were made to obtain medical verification for all reported cancers and to obtain death certificates for deceased subjects. Results: A total of 136 breast cancers were observed among the breast implant patients. External analyses, using general population rates from the Surveillance, Epidemiology and End Results (SEER) program, resulted in 152.2 cases expected and a standardized incidence ratio (SIR) of 0.9 (95% CI 0.8-1.1). A comparable SIR was found for the other plastic surgery patients (SIR = 1.0, 95% CI 0.7-1.2). Internal analyses, directly comparing the implant patients with the other plastic surgery patients, showed a RR of 0.8 (95% CI 0.6-1.1). In neither the external nor internal analyses was there any systematic variation in risk by age or calendar year of initial implant. Risk also did not vary by years of follow-up or by type of implant. Risk was not affected by exclusion of patients who received their implants following surgery for benign breast disease. Although breast tumors tended to be detected at a somewhat later stage among the breast implant than the comparison patients, the difference was not statistically significant, nor was there any significant difference in breast cancer mortality between the two groups. Conclusions: Breast implants do not appear to alter the risk of subsequent breast cancer. C1 NCI, Epidemiol & Biostat Program, Bethesda, MD 20892 USA. NCI, Environm Epidemiol Branch, Bethesda, MD 20892 USA. ABT Associates Inc, Chicago, IL USA. Boston Univ, Dept Epidemiol & Biostat, Boston, MA 02215 USA. US FDA, Ctr Devices & Radiol Hlth, Rockville, MD 20857 USA. RP Brinton, LA (reprint author), NCI, Epidemiol & Biostat Program, Bethesda, MD 20892 USA. RI Brinton, Louise/G-7486-2015 OI Brinton, Louise/0000-0003-3853-8562 NR 38 TC 74 Z9 75 U1 0 U2 1 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD NOV PY 2000 VL 11 IS 9 BP 819 EP 827 DI 10.1023/A:1008941110816 PG 9 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 359EB UT WOS:000089597600007 PM 11075871 ER PT J AU Brownson, RC Alavanja, MCR AF Brownson, RC Alavanja, MCR TI Previous lung disease and lung cancer risk among women (United States) SO CANCER CAUSES & CONTROL LA English DT Article DE asthma; case-control; emphysema; females; lung diseases; lung neoplasms; pneumonia; tuberculosis ID CHLAMYDIA-PNEUMONIAE; NONSMOKING WOMEN; MISSOURI; ASSOCIATION; INFECTIONS; EXPOSURE; TAIWAN; ASTHMA AB Objective: The association between previous lung diseases (PLD) and lung cancer risk has not been studied extensively. We conducted a registry-based case-control study to examine the relation between previous lung diseases and lung cancer among women in Missouri. Methods: Incident cases (n = 676) were identified through the Missouri Cancer Registry for the period 1 January 1993 to 31 January 1994. Controls (n = 700) were selected through drivers' license files and Medicare files. Results: Whether analyzing all respondents or in-person interviews only, elevated effect estimates were noted for several types of PLD. Elevated relative risk estimates were shown for chronic bronchitis (odds ratio [OR] = 1.7; 95% confidence interval [CI] = 1.2-2.3), emphysema (OR = 2.7; 95% CI = 1.8-4.2), pneumonia (OR = 1.6; 95% CI = 1.2-2.0), and for all PLDs combined (OR = 1.5; 95% CI = 1.2-1.9). Analysis of only direct interviews did not show a substantial or consistent pattern of change in relative risk estimates. Because PLDs identified close to the time of cancer diagnosis could conceivably be misdiagnosed, resulting from early lung cancer symptoms, we evaluated the effects on risk estimates of a "latency exclusion" of up to three years. When these exclusions were taken into account, ORs remained statistically significantly elevated only for emphysema. Conclusion: When earlier epidemiologic findings and underlying biological and genetic factors are taken into account, an association between PLD and lung cancer is plausible. C1 St Louis Univ, Sch Publ Hlth, Res Ctr, Dept Community Hlth & Prevent Med, St Louis, MO 63108 USA. NCI, Div Canc Epidemiol & Genet, Rockville, MD USA. RP Brownson, RC (reprint author), St Louis Univ, Sch Publ Hlth, Res Ctr, Dept Community Hlth & Prevent Med, 3663 Lindell Blvd, St Louis, MO 63108 USA. FU NCI NIH HHS [N01-CP7-1096-01, N01-CP7-1096-02] NR 31 TC 54 Z9 56 U1 0 U2 1 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD NOV PY 2000 VL 11 IS 9 BP 853 EP 858 DI 10.1023/A:1008999202040 PG 6 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 359EB UT WOS:000089597600011 PM 11075875 ER PT J AU Chan, JM Pietinen, P Virtanen, M Malila, N Tangrea, J Albanes, D Virtamo, J AF Chan, JM Pietinen, P Virtanen, M Malila, N Tangrea, J Albanes, D Virtamo, J TI Diet and prostate cancer risk in a cohort of smokers, with a specific focus on calcium and phosphorus (Finland) SO CANCER CAUSES & CONTROL LA English DT Article DE calcium; diet; fructose; phosphorus; prostate cancer ID VITAMIN-D METABOLITES; UNITED-STATES; CELL-LINES; SUBSEQUENT DEVELOPMENT; RECEPTOR EXPRESSION; ANDROGEN RECEPTOR; BETA-CAROTENE; TOBACCO USE; IN-VIVO; PROLIFERATION AB Background: Calcium, phosphorus, fructose, and animal protein are hypothesized to be associated with prostate cancer risk, potentially via their influence on 1,25-dihydroxyvitamin D(3). We examined these nutrients and overall diet and prostate cancer risk in the Alpha-Tocopherol Beta-Carotene Cancer Prevention Study (ATBC Study). Materials and methods: The ATBC Study was a randomized 2 x 2 trial of alpha-tocopherol and beta-carotene on lung cancer incidence conducted among Finnish male smokers; 27,062 of the men completed a food-use questionnaire at baseline, and comprise the current study population. There were 184 incident clinical (stage 2-4) prostate cancer cases diagnosed between 1985 and 1993. We used Cox proportional hazards models to examine associations between dietary intakes and prostate cancer. Results: We did not observe significant independent associations for calcium and phosphorus and prostate cancer risk. However, men with lower calcium and higher phosphorus intake had a multivariate relative risk of 0.6 (95% CI 0.3-1.0) compared to men with lower intakes of both nutrients, adjusting for age, smoking, body mass index, total energy, education, and supplementation group. Of the other foods and nutrients examined, none was significantly associated with risk. Discussion: This study provides, at best, only weak evidence for the hypothesis that calcium and phosphorus are independently associated with prostate cancer risk, but suggests that there may be an interaction between these nutrients. C1 Harvard Univ, Sch Publ Hlth, Dept Nutr, Boston, MA 02115 USA. Harvard Univ, Sch Publ Hlth, Dept Epidemiol, Boston, MA 02115 USA. Natl Publ Hlth Inst, Dept Nutr, Helsinki, Finland. NCI, NIH, Bethesda, MD 20892 USA. RP Chan, JM (reprint author), Harvard Univ, Sch Publ Hlth, Dept Nutr, 665 Huntington Ave, Boston, MA 02115 USA. EM jmlchan@hsph.harvard.edu RI Albanes, Demetrius/B-9749-2015 FU NCI NIH HHS [N01-CN-45035, T32-CA-09001] NR 61 TC 80 Z9 80 U1 3 U2 4 PU SPRINGER PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD NOV PY 2000 VL 11 IS 9 BP 859 EP 867 DI 10.1023/A:1008947201132 PG 9 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 359EB UT WOS:000089597600012 PM 11075876 ER PT J AU Boone, CW Stoner, GD Bacus, JV Kagan, V Morse, MA Kelloff, GJ Bacus, JW AF Boone, CW Stoner, GD Bacus, JV Kagan, V Morse, MA Kelloff, GJ Bacus, JW TI Chemoprevention with theaflavins of rat esophageal intraepithelial neoplasia quantitatively monitored by image tile analysis SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID TUMORIGENESIS; CANCER; GREEN; TEA AB The objective of the study was to compare three methods of monitoring the inhibition by dietary theaflavins of N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal intraepithelial neoplasia: the mean the grade, measured by computer-assisted quantitative image the analysis; tumor multiplicity; and mean tumor size. A ''tile" is defined as a small portion of a microscopic image at x40, 87 x 292 Can in size. The computer divided the image of esophageal intraepithelial neoplasia into a grid of contiguous tiles and measured four tissue features within each the based on cytonuclear and tissue architectural changes used by pathologists to diagnose intraepithelial neoplasia, The the grade is defined as the weighted sum of the four feature measurements within a the, the weights being determined by Fisher linear discriminant analysis. The mean the grade of 300 tiles is used to grade rat esophageal intraepithelial neoplasia, NMBA was given s,c,, 0.5 mg/kg, three times a week for 5 weeks, Theaflavins were given in the drinking water at 360 ppm (low dose) and 1200 ppm thigh dose) throughout the experiment. In a given set of four groups of rats, one group received theaflavins alone, one NMBA alone, one NMBA plus low-dose theaflavins, and one NMBA plus high-dose theaflavins, One set of four groups, four rats/ group, was sacrificed at the 15th week and another at the 20th week after starting NMBA; a final set with 15 rats/ group was sacrificed at 25 weeks. At the 15th and 20th weeks, the mean tumor grade was the only variable that responded significantly (P < 0.01) to the low dose of dietary theaflavins, In fact, tumor multiplicity and mean tumor size sometimes showed enhancement at these doses. At the 25th week, when there were 15 instead of 4 rats/group, the mean the grade, tumor multiplicity, and mean tumor size were all significantly (P < 0.01) decreased by both low and high doses of theaflavins, The mean the grade is a more sensitive and reproducible variable than tumor multiplicity and mean tumor size in detecting the chemopreventive effects of theaflavins on intraepithelial neoplasia in the rat esophagus, This suggests that the mean the grade may be a useful intermediate end point for use in human chemoprevention trials. C1 NCI, Chemoprevent Agent Dev Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. Ohio State Univ, Sch Publ Hlth, Div Environm Hlth Sci, Columbus, OH 43210 USA. Bacus Labs, Lombard, IL 60148 USA. RP Stoner, GD (reprint author), Ohio State Univ, Sch Publ Hlth, Div Environm Hlth Sci, CHRI 1148,300 W 10th Ave, Columbus, OH 43210 USA. FU NCI NIH HHS [N01-CN-25486-01, P01 CA-46535] NR 13 TC 3 Z9 3 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD NOV PY 2000 VL 9 IS 11 BP 1149 EP 1154 PG 6 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 373YW UT WOS:000165318500002 PM 11097221 ER PT J AU Slebos, RJC Hoppin, JA Tolbert, PE Holly, EA Brock, JW Zhang, RH Bracci, PM Foley, J Stockton, P McGregor, LM Flake, GP Taylor, JA AF Slebos, RJC Hoppin, JA Tolbert, PE Holly, EA Brock, JW Zhang, RH Bracci, PM Foley, J Stockton, P McGregor, LM Flake, GP Taylor, JA TI K-ras and p53 in pancreatic cancer: Association with medical history, histopathology, and environmental exposures in a population-based study SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID LASER-CAPTURE MICRODISSECTION; TUMOR-SUPPRESSOR GENE; ONCOGENE ACTIVATION; POLYCHLORINATED-BIPHENYLS; MOLECULAR ANALYSIS; EXOCRINE PANCREAS; MUTATIONS; SMOKING; CARCINOMAS; COFFEE AB Pancreatic cancer is a highly fatal cancer with few identified risk factors. Increased risk of pancreatic cancer in tobacco smokers and among diabetic patients is well established, and some reports have suggested associations with coffee consumption and occupational exposure to organochlorines. At present, there is little information regarding the possible association of these risk factors with the known genetic alterations found in pancreatic cancers, such as activation of the K-ras oncogene and inactivation of the p53 tumor suppressor gene. Knowledge of such relationships may help to understand the molecular pathways of pancreatic tumorigenesis. We investigated the association between these molecular defects and risk factors for pancreatic cancer in 61 newly diagnosed patients identified through an ongoing study of pancreatic cancer in the San Francisco Bay Area. Interview information was obtained regarding environmental exposures, medical history, and demographic factors, Serum levels of dichlorodiphenyltrichloroethylene (DDE) and polychlorinated biphenyls were available on a subset of 24 patients. Tumor blocks were located from local hospitals and used for K-ras mutational analysis at codon 12 and for p53 protein immunohistochemistry. The molecular analyses were facilitated through the use of laser capture microdissection, which provides a reliable method to obtain almost pure populations of tumor cells. Mutations in K-ms codon 12 were found in 46 (75%) of 61 pancreatic cancers. A prior diagnosis of diabetes was significantly associated with K-ras negative tumors (P = 0.002, Fisher's exact test). The absence of this mutation was also associated with increased serum levels of DDE, although this association was not statistically significant (P = 0.16, Wilcoxon's test). There was no difference in polychlorinated biphenyl levels between the K-ras wild-type and mutant groups. Immunohistochemical staining for p53 protein did not differ by patient characteristics or clinical history, but significant associations were found with poor glandular differentiation (P = 0.002, chi (2) trend test), severe nuclear atypia (P = 0.0007, chi (2) trend test), and high tumor grade (P = 0.004, chi (2) trend test). Our results are suggestive of the presence of K-ras codon 12 mutation-independent tumorigenesis pathways in patients with prior diabetes and possibly in patients with higher serum levels of DDE, Our results also support a role for the p53 tumor suppressor protein in the maintenance of genomic integrity. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. Emory Univ, Rollins Sch Publ Hlth, Dept Environm & Occupat Hlth, Atlanta, GA 30322 USA. Emory Univ, Rollins Sch Publ Hlth, Dept Biostat, Atlanta, GA 30322 USA. Univ Calif San Francisco, Sch Med, Dept Epidemiol & Biostat, San Francisco, CA USA. Ctr Dis Control, Natl Ctr Environm Hlth, Atlanta, GA 30322 USA. RP Taylor, JA (reprint author), NIEHS, Epidemiol Branch, POB 12233,Maildrop A3-05, Res Triangle Pk, NC 27709 USA. RI Tolbert, Paige/A-5676-2015; OI taylor, jack/0000-0001-5303-6398 FU NCI NIH HHS [R01-CA59706] NR 45 TC 69 Z9 71 U1 0 U2 7 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD NOV PY 2000 VL 9 IS 11 BP 1223 EP 1232 PG 10 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 373YW UT WOS:000165318500012 PM 11097231 ER PT J AU Guan, XY Fu, SB Xia, JC Fang, Y Sham, JST Du, BD Zhou, H Lu, S Wang, BQ Lin, YZ Liang, QW Li, XM Du, B Ning, XM Du, JR Li, P Trent, JM AF Guan, XY Fu, SB Xia, JC Fang, Y Sham, JST Du, BD Zhou, H Lu, S Wang, BQ Lin, YZ Liang, QW Li, XM Du, B Ning, XM Du, JR Li, P Trent, JM TI Recurrent chromosome changes in 62 primary gastric carcinomas detected by comparative genomic hybridization SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID IN-SITU HYBRIDIZATION; OVARIAN-CANCER; SOLID TUMORS; AMPLIFICATION; GENE; ADENOCARCINOMA; EXPRESSION; MUTATIONS; DELETION; STOMACH AB Comparative genomic hybridization (CGH) has been applied to detect recurrent chromosome alterations in 62 primary gastric carcinomas. Several nonrandom chromosomal changes, including gains of 8q (31 cases, 50%), 20q (29 cases, 47%) with a minimum gain region at 20q11.2-q12, 13q (21 cases, 34%) with a minimum gain region at 13q22, and 3q (19 cases, 31%) were commonly observed. The regions most frequently lost included: 19p (23 cases, 37%), 37p (21 cases, 33%), and Ip (14 cases, 23%). High copy number gain (DNA sequence amplification) was detected in 6 cases. Amplification of 8q23-q24.2 and 20q11.2-q12 were observed in 3 cases. Gain of 20q and loss of 19p were confirmed by fluorescence in situ hybridization using corresponding bacterial artificial chromosomes (BAC) clones from those regions. The gain and loss of chromosomal regions identified in this study provide candidate regions involved in gastric tumorigenesis. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Hong Kong, Queen Mary Hosp, Dept Clin Oncol, Hong Kong, Peoples R China. Harbin Med Univ, Dept Biol, Med Genet Lab, Harbin, Peoples R China. Sun Yat Sen Univ Med Sci, Ctr Canc, Guangzhou, Peoples R China. Normal Bethune Univ Med Sci, Teaching Hosp 1, Dept Otolaryngol Head & Neck Surg, Changchun, Peoples R China. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. Harbin Med Univ, Tumor Hosp, Dept Pathol, Harbin, Peoples R China. Harbin Med Univ, Affiliated Hosp 2, Dept Pathol, Harbin, Peoples R China. RP Guan, XY (reprint author), Univ Hong Kong, Queen Mary Hosp, Dept Clin Oncol, Room 129,Professorial Block,Pokfulam Rd, Hong Kong, Peoples R China. RI Guan, Xin-Yuan/A-3639-2009 OI Guan, Xin-Yuan/0000-0002-4485-6017 NR 24 TC 40 Z9 55 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0165-4608 J9 CANCER GENET CYTOGEN JI Cancer Genet. Cytogenet. PD NOV PY 2000 VL 123 IS 1 BP 27 EP 34 DI 10.1016/S0165-4608(00)00306-X PG 8 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 384AA UT WOS:000165915800002 PM 11120330 ER PT J AU Ferro, P Catalano, MG Raineri, M Reato, G dell'Eva, R Risio, M Foa, R Fortunati, N Pfeffer, U AF Ferro, P Catalano, MG Raineri, M Reato, G dell'Eva, R Risio, M Foa, R Fortunati, N Pfeffer, U TI Somatic alterations of the androgen receptor CAG repeat in human colon cancer delineate a novel mutation pathway independent of microsatellite instability SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID BULBAR MUSCULAR-ATROPHY; HUMAN BREAST-CANCER; REPEATED SEQUENCES; GENE-EXPRESSION; PROSTATE-CANCER; DISEASE AB The human androgen receptor gene contains a polymorphic CAG repeat region ranging from 8 to about 35 repeats in the normal human population. The repeat length is inversely related to the transactivation potential of the receptor. We have analyzed the repeat length in 50 sporadic colon cancer samples in comparison to surrounding healthy mucosa and have found somatic reductions of up to 10 repeats in 5 cases (10%), 3 of which were complex, probably involving both alleles. Alterations occurred in tumors with and without microsatellite instability indicating that they follow an independent mutation pathway. The similar repeat of the huntingtin gene did not show any somatic alterations in the same cases. No correlation to sex, tumor stage, location, or histology was evident. In the tumors that showed somatic reductions, the reduced allele was present in at least half of the cells and thus in most, if not all, of the tumor component of the sample. Somatic reductions of the androgen receptor CAG repeat thus occur frequently, through a pathway distinct from microsatellite instability and early during colon carcinogenesis. The receptor is expressed in most normal and neoplastic tissue samples analyzed. Apparent growth selection of cells bearing shortened AR alleles suggests that androgens contribute to colon carcinogenesis in a yet unknown way. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Natl Canc Inst, Mol Biol Lab, Genoa, Italy. Univ Turin, Azienda Osped S Giovanni Battista, UOADU Med Gen 2, Lab Endocrinol, Turin, Italy. Univ Turin, Dipartimento Fisiopatol Clin, Turin, Italy. Univ Turin, Dipartimento Sci Biomed & Oncol Umana, Turin, Italy. Inst Canc Res & Treatment, IRCC, Dept Pathol, Turin, Italy. RP Pfeffer, U (reprint author), Natl Canc Inst, Mol Biol Lab, Genoa, Italy. RI Catalano, Maria Graziella/J-4320-2016; Fortunati, Nicoletta/K-3081-2016; Pfeffer, Ulrich/J-7064-2016 OI Catalano, Maria Graziella/0000-0002-1170-8245; Fortunati, Nicoletta/0000-0001-8069-9621; Pfeffer, Ulrich/0000-0003-0872-4671 NR 22 TC 18 Z9 18 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0165-4608 J9 CANCER GENET CYTOGEN JI Cancer Genet. Cytogenet. PD NOV PY 2000 VL 123 IS 1 BP 35 EP 40 DI 10.1016/S0165-4608(00)00296-X PG 6 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 384AA UT WOS:000165915800003 PM 11120331 ER PT J AU Horig, H Lee, DS Conkright, W Divito, J Hasson, H LaMare, M Rivera, A Park, D Tine, J Guito, K Tsang, KWY Schlom, J Kaufman, HL AF Horig, H Lee, DS Conkright, W Divito, J Hasson, H LaMare, M Rivera, A Park, D Tine, J Guito, K Tsang, KWY Schlom, J Kaufman, HL TI Phase I clinical trial of a recombinant canarypoxvirus (ALVAC) vaccine expressing human carcinoembryonic antigen and the B7.1 co-stimulatory molecule SO CANCER IMMUNOLOGY IMMUNOTHERAPY LA English DT Article DE tumor vaccine; costimulation; poxvirus; carcinoembryonic antigen ID CYTOTOXIC T-LYMPHOCYTE; ADVANCED COLORECTAL-CANCER; COSTIMULATORY MOLECULE; DENDRITIC CELLS; TUMOR REJECTION; PEPTIDE; IDENTIFICATION; VIRUS; B7-2; IMMUNITY AB The generation of cytotoxic effector T cells requires delivery of two signals, one derived from a specific antigenic epitope and one from a costimulatory molecule. A phase I clinical trial was conducted with a non-replicating canarypoxvirus (ALVAC) constructed to express both human carcinoembryonic antigen (CEA) and the B7.1 costimulatory molecule. This was the first study in cancer patients to determine if the delivery of costimulation with a tumor vaccine was feasible and improved immune responses. Three cohorts of six patients, each with advanced CEA-expressing adenocarcinomas, were treated with increasing doses of an ALVAC-CEA-B7.1 vaccine (4.5 x 10(6), 4.5 x 10(7), and 4.5 x 10(8) plaque-forming units, PFU). Patients were vaccinated by intramuscular injection every 4 weeks for 3 months and monitored for side-effects, tumor growth and anti-CEA immune responses. ALVAC-CEA-B7.1 at doses up to 4.5 x 108 PFU was given without evidence of significant toxicity or autoimmune reactions. Three patients experienced clinically stable disease that correlated with increasing CEA-specific precursor T cells, as shown by in vitro interferon-gamma enzyme-linked immunoassay spot tests (ELISPOT). These three patients underwent repeated vaccination resulting in augmented CEA-specific T cell responses. This study represents the first use of costimulation to enhance antitumor vaccines in cancer patients. This approach resulted in CEA-specific immunity associated with stable diseases in three patients. This study also demonstrated that CEA-specific T cell responses could be sustained by repeated vaccinations. Although the number of patients was small, the addition of B7.1 to virus-based vaccines may improve immunological and stable diseases to vaccination against tumor-associated antigens with tolerable toxicity. C1 Albert Einstein Canc Ctr, Bronx, NY 10461 USA. Virogenet Corp, Troy, NY 12180 USA. Pasteur Merieux Connaught Labs, Swiftwater, PA USA. NCI, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. RP Kaufman, HL (reprint author), Albert Einstein Canc Ctr, 1300 Morris Pk Ave, Bronx, NY 10461 USA. FU PHS HHS [R03 80647-01] NR 42 TC 161 Z9 170 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-7004 J9 CANCER IMMUNOL IMMUN JI Cancer Immunol. Immunother. PD NOV PY 2000 VL 49 IS 9 BP 504 EP 514 DI 10.1007/s002620000146 PG 11 WC Oncology; Immunology SC Oncology; Immunology GA 370EX UT WOS:000165111500006 PM 11092617 ER PT J AU Guo, QM Malek, RL Kim, S Chiao, C He, M Ruffy, M Sanka, K Lee, NH Dang, CV Liu, ET AF Guo, QM Malek, RL Kim, S Chiao, C He, M Ruffy, M Sanka, K Lee, NH Dang, CV Liu, ET TI Identification of c-Myc responsive genes using rat cDNA microarray SO CANCER RESEARCH LA English DT Article ID CELL-GROWTH; CYCLIN D1; EXPRESSION; ABSENCE; TARGET AB c-Myc functions; through direct activation or repression of transcription. Using cDNA microarray analysis, we have identified c-Mj c-responsive genes by comparing gene expression profiles between c-myc null and c-myc wildtype rat fibroblast cells and between c-myc null and c-myc nub cells reconstituted with c-myc. From a panel of 4400 cDNA elements, we found 198 genes responsive to c-myc when comparing wild-type or reconstituted cells with the null cells. The plurality of the named c-Myc-responsive genes that were up-regulated, including 30 ribosomal protein genes, are involved in macromolecular synthesis and metabolism, suggesting a major role of c-Myc in the regulation of protein synthetic and metabolic pathways. When ectopically overexpressed, c-Myc induced a different and smaller set of c-Myc-responsive genes as compared with the physiologically expressed c-Myc condition. Thus, these results from expression profiling suggest a new primary function for c-Myc and raise the possibility that the physiological and transforming functions of c-myc may be separable. C1 NCI, Div Clin Sci, Med Branch, Dept Canc & Cell Biol,Mol Signaling & Oncogenesis, Bethesda, MD 20892 USA. Inst Genomic Res, Dept Cellular & Mol Biol, Rockville, MD 20850 USA. Johns Hopkins Univ, Sch Med, Dept Med, Div Hematol, Baltimore, MD 21205 USA. RP Liu, ET (reprint author), NCI, Div Clin Sci, Med Branch, Dept Canc & Cell Biol,Mol Signaling & Oncogenesis, Bldg 31,Room 3A11-MSC2440,Ctr Dr, Bethesda, MD 20892 USA. RI Liu, Edison/C-4141-2008 NR 15 TC 212 Z9 214 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 NOV 1 PY 2000 VL 60 IS 21 BP 5922 EP 5928 PG 7 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300004 PM 11085504 ER PT J AU Vortmeyer, AO Huang, SC Koch, CA Governale, L Dickerman, RD McKeever, PE Oldfield, EH Zhuang, ZP AF Vortmeyer, AO Huang, SC Koch, CA Governale, L Dickerman, RD McKeever, PE Oldfield, EH Zhuang, ZP TI Somatic von Hippel-Lindau gene mutations detected in sporadic endolymphatic sac tumors SO CANCER RESEARCH LA English DT Article ID SUPPRESSOR GENE; RENAL-CARCINOMA; DISEASE; HETEROZYGOSITY; CHROMOSOME-3 AB Endolymphatic sac tumors (ELSTs) occur sporadically or in association with an autosomal dominantly inherited tumor syndrome, von Hippel-Lindau (VHL) disease. In VHL disease, a germline mutation of the VHL tumor suppressor gene is inherited, and loss of function of the wild-type allele occurs through genetic deletion with subsequent development of neoplastic growth. Genetic alterations associated with sporadic ELSTs are less well understood. In this study, we used tissue microdissection to selectively analyze neoplastic cells from four sporadic ELSTs, In two cases, we detected somatic mutations involving VHL gene exons 1 and 2, respectively. Additionally, one of these cases revealed deletion of the VHL gene locus. Two cases did not reveal VHL gene mutation; one of these two cases showed VHL gene deletion. These results suggest that mutations and allelic deletions of the VHL tumor suppressor gene play a role in the tumorigenesis of sporadic ELSTs. C1 NINDS, Mol Pathogenesis Unit, Bethesda, MD 20892 USA. Univ Michigan Hosp, Dept Pathol, Ann Arbor, MI 48109 USA. RP Vortmeyer, AO (reprint author), NINDS, Mol Pathogenesis Unit, Bldg 10,Room 5D37,10 Ctr Dr, Bethesda, MD 20892 USA. RI Koch, Christian/A-4699-2008; OI Koch, Christian/0000-0003-3127-5739; Koch, Christian/0000-0003-0678-1242 FU NCI NIH HHS [CA68545] NR 20 TC 24 Z9 24 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 1 PY 2000 VL 60 IS 21 BP 5963 EP 5965 PG 3 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300013 PM 11085513 ER PT J AU Liu, SH Netzel-Arnett, S Birkedal-Hansen, H Leppla, SH AF Liu, SH Netzel-Arnett, S Birkedal-Hansen, H Leppla, SH TI Tumor cell-selective cytotoxicity of matrix metalloproteinase-activated anthrax toxin SO CANCER RESEARCH LA English DT Article ID PROTECTIVE ANTIGEN; LETHAL FACTOR; FIBROBLAST COLLAGENASE; SEQUENCE SPECIFICITIES; EXTRACELLULAR-MATRIX; ADENYLATE-CYCLASE; EUKARYOTIC CELLS; MAMMALIAN-CELLS; SURFACE; INHIBITORS AB Matrix metalloproteinases (MMPs) are overexpressed in a variety of tumor tissues and cell lines, and their expression is highly correlated to tumor invasion and metastasis. To exploit these characteristics in the design of tumor cell-selective cytotoxins, we constructed two mutated anthrax toxin protective antigen (PA) proteins in which the furin protease cleavage site is replaced by sequences selectively cleaved by MMPs. These MMP-targeted PA proteins were activated rapidly and selectively on the surface of MMP-overexpressing tumor cells. The activated PA proteins caused internalization of a recombinant cytotoxin, FP59, consisting of anthrax toxin lethal factor residues 1-254 fused to the ADP-ribosylation domain of Pseudomonas exotoxin A. The toxicity of the mutated PA proteins for MMP-overexpressing cells was blocked by hydroxamate inhibitors of MMPs, including BB94, and by a tissue inhibitor of matrix metalloproteinases (TIMP-2). The mutated PA proteins killed MMP-overexpressing tumor cells while sparing nontumorigenic normal cells when these were grown together in a coculture model, indicating that PA activation occurred on the tumor cell surface and not in the supernatant, This method of achieving cell-type specificity is conceptually distinct from, and potentially synergistic with, the more common strategy of retargeting a protein toxin by fusion to a growth factor, cytokine, or antibody. C1 Natl Inst Dent & Craniofacial Res, NIH, Oral Infect & Immun Branch, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, NIH, Matrix Metalloproteinase Unit, Bethesda, MD 20892 USA. RP Leppla, SH (reprint author), Natl Inst Dent & Craniofacial Res, NIH, Oral Infect & Immun Branch, 30 Convent Dr,MSC 4350,Bldg 30,Room 316, Bethesda, MD 20892 USA. NR 49 TC 85 Z9 95 U1 1 U2 6 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 1 PY 2000 VL 60 IS 21 BP 6061 EP 6067 PG 7 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300028 PM 11085528 ER PT J AU McDevitt, MR Barendswaard, E Ma, D Lai, L Curcio, MJ Sgouros, G Ballangrud, AM Yang, WH Finn, RD Pellegrini, V Geerlings, MW Lee, M Brechbiel, MW Bander, NH Cordon-Cardo, C Scheinberg, DA AF McDevitt, MR Barendswaard, E Ma, D Lai, L Curcio, MJ Sgouros, G Ballangrud, AM Yang, WH Finn, RD Pellegrini, V Geerlings, MW Lee, M Brechbiel, MW Bander, NH Cordon-Cardo, C Scheinberg, DA TI An alpha-particle emitting antibody ([Bi-213]J591) for radioimmunotherapy of prostate cancer SO CANCER RESEARCH LA English DT Article ID MONOCLONAL-ANTIBODY; MEMBRANE ANTIGEN; ANTI-CD33; PHARMACOKINETICS; EXPRESSION; CONSTRUCTS; DOSIMETRY; TOXICITY; LEUKEMIA; GROWTH AB A novel Lu-particle emitting monoclonal antibody construct targeting the external domain of prostate-specific membrane antigen (PSMA) was prepared and evaluated in vitro and bl vivo, The chelating agent, N-[2-amino-3-(p-isothiocyanatophen-yl)propyl]-trans-cyclohexane-1,2-diamine-N,N',N',N",N"-pentaacetic acid, was appended to J591 monoclonal antibody to stably bind the Bi-213 radiometal ion. Bismuth-213 is a shortlived (t(1/2) = 46 min) radionuclide that emits high energy or-particles with an effective range of 0.07-0.10 mm that are ideally suited to treating single-celled neoplasms and micrometastatic carcinomas. The LNCaP prostate cancer cell line had an estimated 180,000 molecules of PSMA per cell; J591 bound to PSMA with a 3-nM affinity. After binding, the radio-labeled construct-antigen complex was rapidly internalized into the cell, carrying the radiometal inside. [213Bi]J591 was specifically cytotoxic to LNCaP. The LD50 value of [Bi-213]J591 was 220 nCi/ml at a specific activity of 6.4 Ci/g. The potency and specificity of [Bi-213]J591 directed against LNCaP spheroids, an in vitro model for micrometastatic cancer, also was investigated. [Bi-213]J591 effectively stopped growth of LNCaP spheroids relative to an equivalent dose of the irrelevant control [Bi-213]HuM195 or unlabeled J591, Cytotoxicity experiments in vivo were carried out in an athymic nude mouse model with an i.m. xenograft of LNCaP cells. [Bi-213]J591 was able to significantly improve (P < 0.0031) median tumor-free survival (54 days) in these experiments relative to treatment with irrelevant control [Bi-213]HuM195 (33 days), or no treatment (31 days). Prostate-specific antigen (PSA) was also specifically reduced in treated animals. At day 51, mean PSA values were 104 ng/ml +/- 54 ng/ml (n = 4, untreated animals), 66 ng/ml +/- 16 ng/ml (n = 6, animals treated with [Bi-213]HuM195), and 28 ng/ml +/- 22 ng/ml (n = 6, animals treated with [Bi-213]J591). The reduction of PSA levels in mice treated with [Bi-213]J591 relative to mice treated with [Bi-213]HuM195 and untreated control animals was significant with P < 0.007 and P < 0.0136, respectively. In conclusion, a novel [Bi-213]-radiolabeled J591 has been constructed that selectively delivers -particles to prostate cancer cells for potent and specific killing irt vitro and in vivo. C1 Mem Sloan Kettering Canc Ctr, Dept Pharmacol & Mol Therapeut, New York, NY 10021 USA. Mem Sloan Kettering Canc Ctr, Dept Radiol, New York, NY 10021 USA. Mem Sloan Kettering Canc Ctr, Dept Med Phys, New York, NY 10021 USA. Cornell Univ, Weill Med Coll, New York Presbyterian Hosp, Dept Urol, New York, NY 10021 USA. Ludwig Inst Canc Res, New York, NY 10021 USA. NCI, Radiat Oncol Branch, Radioimmune & Inorgan Chem Sect, NIH, Bethesda, MD 20892 USA. Pharmactinium Inc, Chevy Chase, MD 20815 USA. RP Scheinberg, DA (reprint author), Mem Sloan Kettering Canc Ctr, Dept Pharmacol & Mol Therapeut, 1275 York Ave, New York, NY 10021 USA. FU NCI NIH HHS [R01-CA55349] NR 32 TC 155 Z9 158 U1 3 U2 9 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 NOV 1 PY 2000 VL 60 IS 21 BP 6095 EP 6100 PG 6 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300033 PM 11085533 ER PT J AU Amundson, SA Myers, TG Scudiero, D Kitada, S Reed, JC Fornace, AJ AF Amundson, SA Myers, TG Scudiero, D Kitada, S Reed, JC Fornace, AJ TI An informatics approach identifying markers of chemosensitivity in human cancer cell lines SO CANCER RESEARCH LA English DT Article ID DNA-DAMAGING AGENTS; ANTICANCER DRUG SCREEN; GROWTH-ARREST GENES; GAMMA-RAY RESPONSE; GENOTOXIC-STRESS; MESSENGER-RNA; MOLECULAR PHARMACOLOGY; CDNA MICROARRAY; MAMMALIAN-CELLS; LEUKEMIA-CELLS AB We have used a sensitive and reproducible method of measuring mRNA expression to compare basal levels of 10 transcripts in the 60 cell lines of the National Cancer Institute's in vitro anticancer drug screen (NCI-ACDS) under conditions of exponential growth. The strongest correlation among these target genes was between levels of CIP1/WAF1 and BAX. Levels of the three major growth arrest and DNA damage-inducible gene transcripts, (GADD34, GADD45, and GADD153), which are coordinately regulated in response to many stresses, were also correlated across the 60 cell lines. Although the stress induction of several of the transcripts studied here has been shown to be dependent on wild-type p53 status, basal levels of only CIP1/WAF1 and BAX were found to correlate with p53 status. As expected, basal expression of O-6 alkyl guanine alkyl-transferase correlated well with resistance to OG-alkylating agents (r = -0.44) but not with resistance to alkylators with different mechanisms of action (r = -0.04). When basal expression levels of the 10 genes across the NCI-ACDS panel were compared with sensitivities to a panel of 122 standard chemotherapy agents, the most striking relationship was a strong negative correlation (r = -0.3) between basal BCL-X levels and sensitivity to drugs in all of the mechanistic classes except one class of antimetabolites. Sensitivities to a maximally diverse sample of 1200 from 70,000 compounds tested in the NCI-ACDS of agents were also negatively correlated with BCL-X levels. A novel application of factor analysis revealed that the newly discovered associations were independent of previously demonstrated sensitivity factors such as p53 mutation status and native population doubling time. A similar pattern of correlation was seen for Bcl-X-L protein levels. Conversely, BAX and BCL2, two other genes associated with regulation of apoptosis, showed no overall correlation with drug sensitivities. This suggests that BCL-X may play a unique role in general resistance to cytotoxic agents, with the cell lines demonstrating relative resistance to 70,000 cytotoxic agents in the NCI-ACDS being characterized by high BCLX expression. C1 NCI, Biol Res Lab, NIH, Bethesda, MD 20892 USA. NCI, Dev Therapeut Program, NIH, Bethesda, MD 20892 USA. Frederick Canc Res Facil, Frederick, MD 21702 USA. Burnham Inst, La Jolla, CA 92037 USA. RP Amundson, SA (reprint author), NCI, Biol Res Lab, NIH, 37 Convent Dr,Bldg 37,Room 5C09, Bethesda, MD 20892 USA. RI Fornace, Albert/A-7407-2008 OI Fornace, Albert/0000-0001-9695-085X FU NCI NIH HHS [CA78040]; NIGMS NIH HHS [GM60554] NR 51 TC 278 Z9 286 U1 2 U2 14 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 NOV 1 PY 2000 VL 60 IS 21 BP 6101 EP 6110 PG 10 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300034 PM 11085534 ER PT J AU Bowen, C Bubendorf, L Voeller, HJ Slack, R Willi, N Sauter, G Gasser, TC Koivisto, P Lack, EE Kononen, J Kallioniemi, OP Gelmann, EP AF Bowen, C Bubendorf, L Voeller, HJ Slack, R Willi, N Sauter, G Gasser, TC Koivisto, P Lack, EE Kononen, J Kallioniemi, OP Gelmann, EP TI Loss of NKX3.1 expression in human prostate cancers correlates with tumor progression SO CANCER RESEARCH LA English DT Article ID RECEPTOR GENE-EXPRESSION; HOMEOBOX GENE; CHROMOSOME 8P12-21; ALLELIC LOSS; 8P21; ESTROGEN; PROBASIN; REGION AB NKX3.1 is a prostate-specific homeobox gene located on chromosome 8p21, In the mouse, Nkx3.1 has growth-suppressive and differentiating effects on prostatic epithelium. Mutations of the coding region of NKX3.1 were not found in human prostate cancer, failing to support the notion that NKX3.1 was a tumor suppressor gene. To study the expression of NKX3.1 protein in human tissues and prostate cancer, we derived a rabbit antiserum against purified recombinant NKX3.1, Among normal human tissues, NKX3.1 expression was seen in testis, in rare pulmonary mucous glands, and in isolated regions of transitional epithelium of the ureter, NKX3.1 was uniformly expressed in nuclei of normal prostate epithelial cells in 61 histological sections from radical prostatectomy specimens. We analyzed 507 samples of neoplastic prostate epithelium, most of which were contained on a tissue microarray that contained samples from different stages of prostatic neoplasia. We observed complete loss of NKX3.1 expression in 5% of benign prostatic hyperplasias, 20% of high-grade prostatic intraepithelial neoplasias, 6% of T1(a/b) samples, 22% of T3/4 samples, 34% of hormone-refractory prostate cancers, and 78% of metastases, Our data show that NKX3.1 expression is highly, but not exclusively, specific for the prostate. Loss of NKX3.1 expression is strongly associated with hormone-refractory disease and advanced tumor stage in prostate cancer (P < 0.0001). C1 Georgetown Univ, Vincent T Lombardi Canc Res Ctr, Washington, DC 20007 USA. Georgetown Univ, Dept Pathol, Washington, DC 20007 USA. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. Tampere Univ Hosp, Canc Genet Lab, Tampere, Finland. Univ Basel, Inst Pathol, Basel, Switzerland. Univ Basel, Urol Clin, Basel, Switzerland. RP Gelmann, EP (reprint author), Georgetown Univ, Vincent T Lombardi Canc Res Ctr, 3800 Reservoir Rd NW, Washington, DC 20007 USA. RI Kallioniemi, Olli/H-5111-2011; Bubendorfl, Lukas/H-5880-2011; Kallioniemi, Olli/H-4738-2012 OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332 FU NCI NIH HHS [CA78327] NR 21 TC 224 Z9 239 U1 1 U2 6 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD NOV 1 PY 2000 VL 60 IS 21 BP 6111 EP 6115 PG 5 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300035 PM 11085535 ER PT J AU Lerman, MI Minna, JD AF Lerman, MI Minna, JD CA Int Lung Canc Chromosome TI The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3: Identification and evaluation of the resident candidate tumor suppressor genes SO CANCER RESEARCH LA English DT Review ID CELL-LINE U2020; HIPPEL-LINDAU DISEASE; SHORT ARM; LYSOSOMAL HYALURONIDASE; BRONCHIAL EPITHELIUM; EXPRESSION PATTERNS; MOLECULAR ANALYSIS; HUMAN SEMAPHORIN; DNA METHYLATION; ALPHA-SUBUNIT AB We used overlapping and nested homozygous deletions, contig building, genomic sequencing, and physical and transcript mapping to further define a similar to 630-kb lung cancer homozygous deletion region harboring one or more tumor suppressor genes (TSGs) on chromosome 3p21.3, This location was identified through somatic genetic mapping in tumors, cancer cell lines, and premalignant lesions of the lung and breast, including the discovery of several homozygous deletions. The combination of molecular manual methods and computational predictions permitted us to detect, isolate, characterize, and annotate a set of 25 genes that likely constitute the complete set of protein-coding genes residing in this similar to 630-kb sequence. A subset of 19 of these genes was found within the deleted overlap region of similar to 370-kb. This region was further subdivided by a nesting 200-kb breast cancer homozygous deletion into two gene sets: 8 genes lying in the proximal similar to 120-kb segment and 11 genes lying in the distal similar to 250-kb segment. These 19 genes were analyzed extensively by computational methods and were tested by manual methods for loss of expression and mutations in lung cancers to identify candidate TSGs from within this group. Four penes showed loss-of-expression or reduced mRNA levels in non-small cell lung cancer (CACNA2D2/alpha2 delta -2, SEMA3B [formerly SEMA(V) BLU, and HYAL1] or small cell lung cancer (SEMA3B, BLU, and HYAL1) cell lines. We found six of the genes to have two or more amino acid sequence-altering mutations including BLU, NPRL2/Gene21, FUS1, HYAL1, FUS2, and SEMA3B. However, none of the 19 genes tested for mutation showed a frequent (>10%) mutation rate in lung cancer samples. This led us to exclude several of the genes in the region as classical tumor suppressors for sporadic lung cancer. On the other hand, the putative lung cancer TSG in this location may either be inactivated by tumor-acquired promoter hypermethylation or belong to the novel class of haploinsufficient genes that predispose to cancer in a hemizygous (+/-) state hut do not show a second mutation in the remaining wild-type allele in the tumor. We discuss the data in the context of novel and classic cancer gene models as applied to lung carcinogenesis. Further functional testing of the critical genes by gene transfer and gene disruption strategies should permit the identification of the putative lung cancer TSG(s), LUCA. Analysis of the similar to 630-kb sequence also provides an opportunity to probe and understand the genomic structure, evolution; and functional organization of this relatively gene-rich region. C1 NCI, FCRDC, Immunobiol Lab, Frederick, MD 21702 USA. Univ Texas, SW Med Ctr, Hamon Ctr Therapeut Oncol Res, Dallas, TX 75390 USA. RP Minna, JD (reprint author), NCI, FCRDC, Immunobiol Lab, Frederick, MD 21702 USA. EM John.Minna@UTSouthwestern.edu RI Zabarovsky, Eugene/A-6645-2010 FU NCI NIH HHS [CA71618, N01-CO-56000, P50 CA70907] NR 131 TC 418 Z9 464 U1 1 U2 27 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA 615 CHESTNUT ST, 17TH FLOOR, PHILADELPHIA, PA 19106-4404 USA SN 0008-5472 EI 1538-7445 J9 CANCER RES JI Cancer Res. PD NOV 1 PY 2000 VL 60 IS 21 BP 6116 EP 6133 PG 18 WC Oncology SC Oncology GA 372JJ UT WOS:000165230300036 PM 11085536 ER PT J AU Poetschke, HL Klug, DB Perkins, SN Wang, TTY Richie, ER Hursting, SD AF Poetschke, HL Klug, DB Perkins, SN Wang, TTY Richie, ER Hursting, SD TI Effects of calorie restriction on thymocyte growth, death and maturation SO CARCINOGENESIS LA English DT Article ID DIETARY RESTRICTION; FOOD RESTRICTION; CELLULAR PROLIFERATION; V(D)J RECOMBINATION; P53-DEFICIENT MICE; PRONE MICE; MOUSE; TUMORIGENESIS; LYMPHOCYTES; EXPRESSION AB We previously reported that calorie restriction (CR) significantly delays the spontaneous development of thymic lymphomas and other neoplasms in p53-deficient mice and their wild-type littermates. The purpose of the present study was to further characterize the anti-lymphoma effects of CR by assessing thymocyte growth, death and maturation in response to acute (6 day) and chronic (28 day) CR regimens. Male C57BL/6J mice fed a CR diet (restricted to 60% of control nn libitum intake) for 6 days displayed a severe reduction in thymic size and cellularity, as well as a decrease in splenic size and cellularity; these declines were sustained through 28 days of CR, Mice maintained on a CR diet for 28 days also displayed a significant depletion in the cell numbers of all four major thymocyte subsets defined by CD4 and CD8 expression. Analysis within the immature CD4(-)8(-) thymocyte subset further revealed an alteration in normal CD44 and CD25 subset distribution. in particular, CR for 28 days resulted in a significant decrease in the percentage of the proliferative CD44(-)25(-) subset. In addition, a significant increase in the percentage of the early, pro-T cell CD44(+)25(-) population was detected, indicative of a CR-induced delay in thymocyte maturation. Taken together, these findings suggest that CR suppresses (through several putative mechanisms) lymphomagenesis by reducing the pool of immature thymocytes that constitute the lymphoma-susceptible subpopulation. C1 Univ Texas, MD Anderson Canc Ctr, Dept Carcinogenesis, Smithville, TX 78957 USA. NCI, Frederick Canc Res & Dev Ctr, Basic Res Lab, Frederick, MD 21702 USA. USDA ARS, BHNRC, Phytonutr Lab, Beltsville, MD 20705 USA. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Hursting, SD (reprint author), NCI, Off Prevent Oncol, 6130 Execut Blvd,MSC 7105, Bethesda, MD 20892 USA. FU NCI NIH HHS [P30 CA 16672-21]; NIEHS NIH HHS [P30 ES07784-01] NR 34 TC 22 Z9 24 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD NOV PY 2000 VL 21 IS 11 BP 1959 EP 1964 DI 10.1093/carcin/21.11.1959 PG 6 WC Oncology SC Oncology GA 377ZF UT WOS:000165555800005 PM 11062154 ER PT J AU Chang, RL Wood, AW Kumar, S Lehr, RE Shirai, N Jerina, DM Conney, AH AF Chang, RL Wood, AW Kumar, S Lehr, RE Shirai, N Jerina, DM Conney, AH TI Tumorigenicity of four optically active bay-region 3,4-diol 1,2-epoxides and other derivatives of the nitrogen heterocycle dibenz[c,h]acridine on mouse skin and in newborn mice SO CARCINOGENESIS LA English DT Article ID DIASTEREOMERIC BENZOPYRENE 7,8-DIOL-9,10-EPOXIDES; TUMOR-INITIATING ACTIVITY; RAT-LIVER ENZYMES; DIOL-EPOXIDES; MAMMALIAN-CELLS; MARKED DIFFERENCES; STEREOSELECTIVE METABOLISM; MUTAGENICITY; ENANTIOMERS; 3,4-DIOL-1,2-EPOXIDES AB The nitrogen heterocycle dibenz[c,h]acridine (DB[c,h]ACR) and the enantiomers of the diastereomeric pair of bay-region 3,4-diol 1,2-epoxides as well as other bay-region epoxides and dihydrodiol derivatives of this hydrocarbon have been evaluated for tumorigenicity on mouse skin and in the newborn mouse. On mouse skin, a single topical application of 50 or 200 nmol of compound was followed 10 days later by twice-weekly applications of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate for 20 weeks. DB[c,h]ACR and the four optically pure, bay-region 3,4-diol-1,2-epoxide isomers all had significant tumor-initiating activity, The isomer with (1R,2S,3S,4R) absolute configuration [(+)-DE-2] was the most active diol epoxide isomer, The (-)-(3R,3R)-dihydrodiol of DB[c,h]ACR, the expected metabolic precursor of the bay-region (+)-DE-2, was 4- to 6-fold more tumorigenic than its corresponding (+)-enantiomer, In tumorigenicity studies in newborn mice, a total dose of 70-175 nmol of DB[c,h]ACR or one of its derivatives was Injected i,p, on days 1, 8 and 15 of life, and tumorigenic activity was determined when the mice were 36-39 weeks old. DB[c,h]ACR produced a significant number of pulmonary tumors and also produced hepatic tumors in male mice. Of the four optically active bayregion diol epoxides, only (+)-DE-2 and (+)-DE-1 with (1R,2S,3S,4R) and (1S,2R,3S,4R) absolute configuration, respectively, produced a significant tumor incidence. At an equivalent dose, the (+)-DE-2 isomer produced severalfold more pulmonary tumors and hepatic tumors than the (+)-DE-1 isomer, The (-)-(3R,4R)-dihydrodiol, metabolic precursor of the bay-region (+)-DE-2, was strongly active and induced an equal number of pulmonary and hepatic tumors as did DB[c,lt]ACR. The (+)-(35,4S) dihydrodiol was less active. The bay-region (+)-(1R,2S)-epoxide of 1,2,3,4-tetrahydro DB[c,h]ACR was strongly tumorigenic in newborn mice whereas its (-)-(1S,2R)-enantiomer was inactive. This contrasts with the data on mouse skin where both enantiomers had substantial tumorigenic activity. In summary, the bay-region (+)-(1R,2S,3S,4R)-3,4-diol 1,2-epoxide of DB[c,h]ACR was the most tumorigenic of the four optically active bay-region diol epoxides of DB[c,h]ACR on mouse skin and in the newborn mouse, These results with a nitrogen heterocycle are similar to earlier data indicating high tumorigenic activity for the R,S,S,R bay-region diol epoxides of several carbocyclic polycyclic aromatic hydrocarbons. C1 Rutgers State Univ, Coll Pharm, Canc Res Lab, Piscataway, NJ 08854 USA. Hoffmann La Roche Inc, Roche Res Ctr, Nutley, NJ 07110 USA. SUNY Coll Buffalo, Environm Res Ctr, Div Environm Toxicol & Chem, Buffalo, NY 14222 USA. Univ Oklahoma, Dept Chem & Biochem, Norman, OK 73019 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. RP Chang, RL (reprint author), Rutgers State Univ, Coll Pharm, Canc Res Lab, 164 Frelinghuysen Rd, Piscataway, NJ 08854 USA. FU NCI NIH HHS [CA49756] NR 48 TC 11 Z9 11 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD NOV PY 2000 VL 21 IS 11 BP 1997 EP 2003 DI 10.1093/carcin/21.11.1997 PG 7 WC Oncology SC Oncology GA 377ZF UT WOS:000165555800011 PM 11062160 ER PT J AU Huang, J Hu, N Goldstein, AM Emmert-Buck, MR Tang, ZZ Roth, MJ Wang, QH Dawsey, SM Han, XY Ding, T Li, G Giffen, C Taylor, PR AF Huang, J Hu, N Goldstein, AM Emmert-Buck, MR Tang, ZZ Roth, MJ Wang, QH Dawsey, SM Han, XY Ding, T Li, G Giffen, C Taylor, PR TI High frequency allelic loss on chromosome 17p13.3-p11.1 in esophageal squamous cell carcinomas from a high incidence area in northern China SO CARCINOGENESIS LA English DT Article ID TUMOR-SUPPRESSOR GENE; LASER CAPTURE MICRODISSECTION; SHANXI-PROVINCE; FAMILY HISTORY; CANCER; HETEROZYGOSITY; ALLELOTYPE; EXPRESSION; SITES; HIC-1 AB Allelic loss on chromosome 17p has been reported frequently in esophageal squamous cell carcinoma (ESCC) and generally encompasses the p53 locus at 17p13,1, However, a good correlation between allelic loss on 17p and mutation of p53 has not been found. This suggests the possibility that unknown tumor suppressor genes near p53 may be involved in the development of ESCC, To evaluate this possibility, we analyzed 30 microsatellite markers covering the entire short arm of chromosome 17 in 56 ESCC patients from a high risk population in northern China, including 34 with a family history of upper gastrointestinal (UGI) cancer and 22 without a family history of any cancer. Cancer lifestyle risk factors and clinical/pathological characteristics were also collected. We found frequent allelic loss (greater than or equal to 65%) at 28 of the 30 markers evaluated in these ESCC patients. The highest frequencies of allelic loss (greater than or equal to 80%) were found in three smaller regions: deletion region I located at 17p13,3-p13.2 (between D17S849 and D17S1828); deletion region II located at 17p13,2-p13,1 (between D13S938 and TP53); deletion region III located at 17p13,1-p12 (between D17S804 and D17S799), A number of genes have already been identified in these deleted regions, including: OVCA1, OVCA2 and HIC-1 in deletion region I; p53 in deletion region II; ZNF18, ZNn9, ALDH3 and ALDH10 in deletion region III. These results will help us direct future testing of candidate genes and narrow the search region for major new tumor suppressor genes that may play a role in the pathogenesis of ESCC. C1 Chinese Acad Med Sci, Canc Inst & Hosp, Beijing 100021, Peoples R China. NCI, NIH, Bethesda, MD 20892 USA. Shanxi Canc Hosp, Shanxi 030013, Peoples R China. Informat Management Serv Inc, Silver Spring, MD 20904 USA. RP Taylor, PR (reprint author), Chinese Acad Med Sci, Canc Inst & Hosp, Beijing 100021, Peoples R China. NR 29 TC 30 Z9 34 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD NOV PY 2000 VL 21 IS 11 BP 2019 EP 2026 DI 10.1093/carcin/21.11.2019 PG 8 WC Oncology SC Oncology GA 377ZF UT WOS:000165555800014 PM 11062163 ER PT J AU Haak, LL Grimaldi, M Russell, JT AF Haak, LL Grimaldi, M Russell, JT TI Mitochondria in myelinating cells: calcium signaling in oligodendrocyte precursor cells SO CELL CALCIUM LA English DT Review ID INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR; RAT OPTIC-NERVE; PERMEABILITY TRANSITION PORE; ENDOPLASMIC-RETICULUM; CA2+ SIGNALS; IN-VIVO; ELECTRICAL-ACTIVITY; TRANSDUCTION SYSTEMS; ELEMENTARY EVENTS; WAVE-PROPAGATION C1 NICHD, LCMN, Sect Cell Biol & Signal Transduct, NIH, Bethesda, MD 20892 USA. RP Russell, JT (reprint author), NICHD, LCMN, Sect Cell Biol & Signal Transduct, NIH, Bldg 49,Room 5A-18,49 Convent Dr, Bethesda, MD 20892 USA. EM james@helix.nih.gov RI Haak, Laurel/C-4986-2008; OI Haak, Laurel/0000-0001-5109-3700; Grimaldi, Maurizio/0000-0002-7331-7055 NR 107 TC 15 Z9 15 U1 0 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0143-4160 EI 1532-1991 J9 CELL CALCIUM JI Cell Calcium PD NOV-DEC PY 2000 VL 28 IS 5-6 BP 297 EP 306 DI 10.1054/ceca.2000.0176 PG 10 WC Cell Biology SC Cell Biology GA 392JT UT WOS:000166408500003 PM 11115369 ER PT J AU Marcu, M Bertolotti, A Ron, D Neckers, L AF Marcu, M Bertolotti, A Ron, D Neckers, L TI Hsp90 stabilizes IREI: a transmembrane kinase that mediates the unfolded protein response SO CELL STRESS & CHAPERONES LA English DT Meeting Abstract C1 NCI, NIH, Rockville, MD 20850 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU CELL STRESS SOC INTERNATIONAL PI STORRS PA UNIV CONNECTICUT, DEPT M C B, 75 NORTH EAGLEVILLE RD, U-44, STORRS, CT 06269-3044 USA SN 1355-8145 J9 CELL STRESS CHAPERON JI Cell Stress Chaperones PD NOV PY 2000 VL 5 IS 5 BP 492 EP 492 PG 1 WC Cell Biology SC Cell Biology GA 388DV UT WOS:000166162900041 ER PT J AU Xu, W Mimnaugh, E Rosser, M Nicchitta, C Marcu, M Yarden, Y Neckers, L AF Xu, W Mimnaugh, E Rosser, M Nicchitta, C Marcu, M Yarden, Y Neckers, L TI Divergent sensitivity of ErbB1 and ErbB2 to geldanamycin may be regulated by chaperone proteins SO CELL STRESS & CHAPERONES LA English DT Meeting Abstract C1 NCI, NIH, Rockville, MD 20850 USA. RI YARDEN, YOSEF/K-1467-2012 NR 0 TC 0 Z9 0 U1 0 U2 0 PU CELL STRESS SOC INTERNATIONAL PI STORRS PA UNIV CONNECTICUT, DEPT M C B, 75 NORTH EAGLEVILLE RD, U-44, STORRS, CT 06269-3044 USA SN 1355-8145 J9 CELL STRESS CHAPERON JI Cell Stress Chaperones PD NOV PY 2000 VL 5 IS 5 BP 492 EP 492 PG 1 WC Cell Biology SC Cell Biology GA 388DV UT WOS:000166162900043 ER PT J AU Bristol, JA Orsini, C Lindinger, P Thalhamer, J Abrams, SI AF Bristol, JA Orsini, C Lindinger, P Thalhamer, J Abrams, SI TI Identification of a ras oncogene peptide that contains both CD4(+) and CD8(+) T cell epitopes in a nested configuration and elicits both T cell subset responses by peptide or DNA immunization SO CELLULAR IMMUNOLOGY LA English DT Article DE mouse; CD4(+) and CD8(+) T cells; nested epitopes; peptides; ras oncogenes ID MUTATED RAS; CLASS-I; OVERLAPPING EPITOPES; CANCER; LYMPHOCYTES; VACCINES; CLONES; GENES; PROTOONCOGENES; IMMUNOTHERAPY AB Mutations in ras proto oncogenes are commonly found in a diversity of malignancies and may encode unique, non-self epitopes for T cell-mediated antitumor activity. In a BALB/c (H-2(d)) murine model, we have identified a single peptide sequence derived from the ras oncogenes that contained both CD8(+) and CD4(+) T cell. epitopes in a nested configuration. This peptide reflected ras sequence 4-16, and contained the substitution of Gly to Val at position 12 {i.e., 4-16(Val12)}. Mice immunized with this 13-mer peptide induced a strong antigen (Ag)-specific CD4(+) proliferative response in vitro. In contrast, mice inoculated with the wild-type ras sequence failed to generate a peptide-specific T cell. response, Additionally, mice immunized with the ras 4-16(Val12) peptide concomitantly displayed an Ag-specific CD8(+) cytotoxic T lymphocyte (CTL) response, as determined by lysis of syngeneic tumor target cells incubated with the nominal 9-mer nested epitope peptide {i.e., 4-12(Val12)}, as well as lysis of tumor target cells expressing the corresponding ras codon 12 mutation, Analysis of the V alpha- and V beta -chains of the T cell receptor (TCR) expressed by these CTL revealed usage of the V alpha1 and V beta9 subunits, consistent with the TCR phenotype of anti-ras Val12 CTL lines produced by in vivo immunization with the nominal peptide epitope alone. Moreover, immunization with the nested epitope peptide, as compared to immunization with either the 9-mer CTL peptide alone or an admixture of the 9-mer CTL peptide with an overlapping 13-mer CD4(+) T cell helper peptide {i.e., 5-17(Val12)} lacking the class I N-terminus anchor site, enhanced the production of the CD8(+) T cell response. Finally, immunization with plasmid DNA encoding the ras 4-16(Val12) sequence led to the induction of both Ag-specific proliferative and cytotoxic responses. Overall, these results suggested that a single peptide immunogen containing nested mutant ras-specific CD4(+) and CD8(+) T cell epitopes: (1) can be processed in vivo to induce both subset-specific T lymphocyte responses; and (2) leads to the generation of a quantitatively enhanced CD8(+) CTL response, likely due to the intimate coexistence of CD4(+) help, which may have implications in peptide- or DNA-based immunotherapies. (C) 2000 Academic Press. C1 NCI, Tumor Immunol & Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Salzburg Univ, Inst Chem & Biochem, A-5020 Salzburg, Austria. RP Abrams, SI (reprint author), NCI, Tumor Immunol & Biol Lab, Div Basic Sci, NIH, Bldg 10,Room 5B46,10 Ctr Dr, Bethesda, MD 20892 USA. RI Thalhamer, Josef/E-5787-2011 OI Thalhamer, Josef/0000-0003-2285-6400 NR 39 TC 25 Z9 27 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0008-8749 J9 CELL IMMUNOL JI Cell. Immunol. PD NOV 1 PY 2000 VL 205 IS 2 BP 73 EP 83 DI 10.1006/cimm.2000.1712 PG 11 WC Cell Biology; Immunology SC Cell Biology; Immunology GA 383NX UT WOS:000165889800001 PM 11104579 ER PT J AU Callicott, JH Bertolino, A Mattay, VS Langheim, FJP Duyn, J Coppola, R Goldberg, TE Weinberger, DR AF Callicott, JH Bertolino, A Mattay, VS Langheim, FJP Duyn, J Coppola, R Goldberg, TE Weinberger, DR TI Physiological dysfunction of the dorsolateral prefrontal cortex in schizophrenia revisited SO CEREBRAL CORTEX LA English DT Review ID SPATIAL WORKING-MEMORY; POSITRON-EMISSION-TOMOGRAPHY; CEREBRAL BLOOD-FLOW; MAGNETIC-RESONANCE SPECTROSCOPY; ATTENUATED FRONTAL ACTIVATION; AUTOMATED IMAGE REGISTRATION; NEUROLEPTIC-NAIVE PATIENTS; ANTERIOR CINGULATE CORTEX; VERBAL FLUENCY TASK; CARD SORTING TEST AB Evidence implicates subtle neuronal pathology of the prefrontal cortex (PFC) in schizophrenia, but how this pathology is reflected in physiological neuroimaging experiments remains controversial. We investigated PFC function in schizophrenia using functional magnetic resonance imaging (fMRI) and a parametric version of the n-back working memory (WM) task. In a group of patients who performed relatively well on this task, there were three fundamental deviations from the 'healthy' pattern of PFC fMRI activation to varying WM difficulty. The first characteristic was a greater magnitude of PFC fMRI activation in the context of slightly impaired WM performance (i.e. physiological inefficiency). The second was that the significant correlations between behavioral WM performance and dorsal PFC fMRI activation were in opposite directions in the two groups. Third, the magnitude of the abnormal dorsal PFC fMRI response was predicted by an assay of N-acetylaspartate concentrations (NAA) in dorsal PFC, a measure of neuronal pathology obtained using proton magnetic resonance spectroscopy. Patients had significantly lower dorsal PFC NAA than controls and dorsal PFC NAA inversely predicted the fMRI response in dorsal PFC (areas 9, 46) to varying WM difficulty - supporting the assumption that abnormal PFC responses arose from abnormal PFC neurons. These data suggest that under certain conditions the physiological ramifications of dorsal PFC neuronal pathology in schizophrenia includes exaggerated and inefficient cortical activity, especially of dorsal PFC. C1 NIMH, Clin Brain Disorders Branch, IRP, NIH, Bethesda, MD 20892 USA. NIH, Lab Diagnost Radiol Res, CC, Bethesda, MD 20892 USA. RP Callicott, JH (reprint author), NIMH, Clin Brain Disorders Branch, IRP, NIH, 10 Ctr Dr,Rm 4D-20,MSC 1389, Bethesda, MD 20892 USA. RI Duyn, Jozef/F-2483-2010; Callicott, Joseph/C-9102-2009; Bertolino, Alessandro/O-6352-2016 OI Callicott, Joseph/0000-0003-1298-3334; Bertolino, Alessandro/0000-0002-1251-1380 NR 119 TC 521 Z9 532 U1 9 U2 52 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 1047-3211 J9 CEREB CORTEX JI Cereb. Cortex PD NOV PY 2000 VL 10 IS 11 BP 1078 EP 1092 DI 10.1093/cercor/10.11.1078 PG 15 WC Neurosciences SC Neurosciences & Neurology GA 366DR UT WOS:000089990000003 PM 11053229 ER PT J AU Matveev, V Wang, XJ AF Matveev, V Wang, XJ TI Differential short-term synaptic plasticity and transmission of complex spike trains: to depress or to facilitate? SO CEREBRAL CORTEX LA English DT Article ID NEOCORTICAL PYRAMIDAL NEURONS; INDIVIDUAL HIPPOCAMPAL SYNAPSES; DUAL INTRACELLULAR-RECORDINGS; NEUROTRANSMITTER RELEASE; TRANSMITTER RELEASE; EXCITATORY SYNAPSES; VISUAL-CORTEX; FRACTAL CHARACTER; QUANTAL ANALYSIS; RAT HIPPOCAMPUS AB Experimental studies have revealed conspicuous short-term facilitation and depression that are expressed differentially at distinct classes of cortical synapses. To explore computational implications of synaptic dynamics, we investigated transmission of complex spike trains through a stochastic model of cortical synapse endowed with short-term facilitation and vesicle depletion. Inputs to the synapse model were either real spike train data recorded from the visual and prefrontal cortices of behaving monkeys, or were generated numerically with prescribed temporal statistics. We tested the hypothesis that short-term facilitation could enable synapses to filter out single spikes and favor bursts of action potentials. We found that the ratio between release probabilities for a burst spike and an isolated spike grows monotonically with increasing number of spikes per burst, and with increasing interval between isolated spikes. Burst detection is optimal when the facilitation time constant matches the average burst duration. Using fractal-like spike patterns characterized by long-term power-law temporal correlations and similar to those seen in sensory neurons, we found that facilitation increases correlation at short time scales. In contrast, depression leads to a dramatic reduction in temporal correlations at all time scales, and to a flat ('whitened') power spectrum, thereby decorrelating natural input signals. C1 Brandeis Univ, Volen Ctr Complex Syst, Waltham, MA 02454 USA. NIDDK, Math Res Branch, NIH, Bethesda, MD 20892 USA. RP Wang, XJ (reprint author), Brandeis Univ, Volen Ctr Complex Syst, Waltham, MA 02454 USA. RI Wang, Xiao-Jing/D-2722-2009 FU NIMH NIH HHS [MH53717-01] NR 100 TC 31 Z9 31 U1 0 U2 4 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 1047-3211 J9 CEREB CORTEX JI Cereb. Cortex PD NOV PY 2000 VL 10 IS 11 BP 1143 EP 1153 DI 10.1093/cercor/10.11.1143 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 366DR UT WOS:000089990000008 PM 11053234 ER PT J AU Baird, AE Lovblad, KO Dashe, JF Connor, A Burzynski, C Schlaug, G Straroselskaya, I Edelman, RR Warach, S AF Baird, AE Lovblad, KO Dashe, JF Connor, A Burzynski, C Schlaug, G Straroselskaya, I Edelman, RR Warach, S TI Clinical correlations of diffusion and perfusion lesion volumes in acute ischemic stroke SO CEREBROVASCULAR DISEASES LA English DT Article DE diffusion-weighted imaging; perfusion; magnetic resonance; ischemic stroke; cerebrovascular disorders; stroke outcome ID ACUTE CEREBRAL INFARCTION; WEIGHTED MRI AB The aim of this study was to describe the clinico-radiological correlations of magnetic resonance (MR) perfusion and diffusion-weighted imaging (DWI) abnormalities in ischemic stroke. Eighteen patients had undergone MR imaging and clinical evaluation within 24 h of symptom onset and at or after 7 days. During the first 24 h the volume of perfusion abnormality (measured on the relative mean transit time map) was larger than the DWI lesion in 12/18 patients. In 6/18 patients the DWI lesion volume was larger. Acutely (<24 h) all lesion volumes showed a significant correlation with acute clinical severity measured by the National Institutes of Health Stroke Scale score. The correlations of the hypoperfusion volume (rho = 0.86, p = 0.0001) and the volume 'tissue at risk' (larger than the DWI and perfusion lesion volumes, rho = 0.86, p = 0.0001) with acute clinical severity were slightly higher than for the DWI lesion volume (rho = 0.76, p = 0.0001). The difference between the volume of tissue at risk (acutely) and the infarct on follow-up T-2-weighted imaging correlated significantly with change in clinical severity from acute to chronic time points (rho = 0.72, p = 0.001). Such clinico-radiological relationships may support the use of DWI and perfusion MR in decisions concerning the administration and evaluation of stroke therapies. Copyright (C) 2000 S. Karger AG, Basel. C1 NINDS, Div Diagnost & Therapeut, NIH, Bethesda, MD 20892 USA. Beth Israel Deaconess Med Ctr, Dept Neurol, Boston, MA 02215 USA. Beth Israel Deaconess Med Ctr, Dept Radiol, Boston, MA 02215 USA. Harvard Univ, Sch Med, Boston, MA USA. RP Baird, AE (reprint author), NINDS, Div Diagnost & Therapeut, NIH, 36 Convent Dr,MSC 4129,Room 4A03, Bethesda, MD 20892 USA. NR 33 TC 52 Z9 54 U1 0 U2 1 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 1015-9770 J9 CEREBROVASC DIS JI Cerebrovasc. Dis. PD NOV-DEC PY 2000 VL 10 IS 6 BP 441 EP 448 DI 10.1159/000016105 PG 8 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA 373ND UT WOS:000165294800005 PM 11070374 ER PT J AU Kadiiska, MB De Costa, KS Mason, RP Mathews, JM AF Kadiiska, MB De Costa, KS Mason, RP Mathews, JM TI Reduction of 1,3-diphenyl-1-triazene by rat hepatic microsomes, by cecal microflora, and in rats generates the phenyl radical metabolite: An ESR spin-trapping investigation SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID ARENEDIAZONIUM IONS; FISCHER-344 RATS; IN-VITRO; PHENYLHYDRAZINE; ZIDOVUDINE; ADDUCTS; INVIVO; TRAPS; EPR; DNA AB An ESR spin-trapping technique was used to determine whether free radical metabolites are formed as a result of the reduction of 1,3-diphenyl-1-triazene (DPT) in vivo and in vitro by components of the cytochrome P450 (P450) mixed-function oxidase system in microsomes or by gut microflora in anaerobic cecal incubations. The ESR spectrum of the DMPO-phenyl radical adduct was detected in a microsomal incubation containing DPT, DMPO, and NADPH with the following hyperfine coupling constants: alpha (N) = 15.95 G and alpha (H)(beta) = 24.37 G. The amplitude of the spectrum from the phenyl radical adduct generated in microsomal incubations of DPT with DMPO and NADPH was not attenuated by the P450 inhibitor 1-aminobenzotriazole (ABT) or by carbon monoxide, indicating that P450 is not significantly involved in phenyl radical formation. The formation of a DMPO-phenyl radical adduct was also catalyzed by recombinant human cytochrome P450 reductase. Addition of anti-rat P450 reductase antibody led to an attenuation of the signal in incubations containing either microsomes or reductase. Low concentrations of DMPO-phenyl radical adducts were detected by ESR in the toluene extract of cecal contents containing DPT and the spin trap. In the in vivo experiments with rats treated with DPT and the spin trap DMPO, the six-line ESR signal of the DMPO-phenyl radical adduct was readily detected in bile 40-60 min after rats were treated with DPT and DMPO. The results show for the first time that the phenyl radical is formed by the reduction of DPT and may indicate a toxic potential for this chemical. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. Res Triangle Inst, Ctr Bioorgan Chem, Res Triangle Pk, NC 27709 USA. RP Kadiiska, MB (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 26 TC 11 Z9 11 U1 0 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD NOV PY 2000 VL 13 IS 11 BP 1082 EP 1086 DI 10.1021/tx000098v PG 5 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 376ML UT WOS:000165461300002 PM 11087429 ER PT J AU Kadiiska, MB Mason, RP AF Kadiiska, MB Mason, RP TI Ethylene glycol generates free radical metabolites in rats: An ESR in vivo spin trapping investigation SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID CALCIUM-OXALATE NEPHROLITHIASIS; LIPID-PEROXIDATION; AQUEOUS-SOLUTION; IN-VIVO; TOXICITY; OXIDATION; ACID; 4-METHYLPYRAZOLE; FORMALDEHYDE; MICROSOMES AB Ethylene glycol, best known as antifreeze, is most often ingested accidentally or as a substitute for alcochol by chronic alcohol abusers. The toxicity of ethylene glycol poisoning is due to its toxic metabolites rather than to ethylene glycol itself. In this study, electron spin resonance (ESR) spectroscopy has been used to study free radical generation in rats by acute ethylene glycol poisoning. The radical spin trapping technique was applied where the spin trapping agent alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) reacted with free radical metabolites to form radical adducts in vivo. The radical adducts from ethylene glycol intoxication were detected in both the bile and urine samples of male Sprague-Dawley rats. The identification of the POBN-(.)[C-13]ethylene glycol radical adduct provides for the first time direct ESR evidence for the generation of the ethylene glycol-derived radicals during acute intoxication by ethylene glycol, suggesting a new metabolic pathway. Simultaneous administration of alcohol dehydrogenase inhibitor 4-methylpyrazole with ethylene glycol resulted in an enhanced free radical generation in the bile. This report is the first evidence of ethylene glycol free radical metabolism in rats with acute ethylene glycol intoxication. C1 NIEHS, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. RP Kadiiska, MB (reprint author), NIEHS, Lab Pharmacol & Chem, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 36 TC 14 Z9 14 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD NOV PY 2000 VL 13 IS 11 BP 1187 EP 1191 DI 10.1021/tx9901940 PG 5 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 376ML UT WOS:000165461300015 PM 11087442 ER PT J AU Szeliga, J Amin, S AF Szeliga, J Amin, S TI Quantitative reactions of anti 5,9-dimethylchrysene dihydrodiol epoxide with DNA and deoxyribonucleotides SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article DE DNA adducts; 5,9-dimethylchrysene; polycyclic aromatic hydrocarbon dihydrodiol epoxides ID COVALENT NUCLEOSIDE ADDUCTS; REGION 3,4-DIOL 1,2-EPOXIDES; 11,12-DIHYDRODIOL 13,14-EPOXIDE; CYTOSINE BASES; 5,6-DIMETHYLCHRYSENE; TUMORIGENICITY; ADENINE; GUANINE AB Native as well as denatured calf thymus DNA, deoxyguanylic and deoxyadenylic acid, respectively, were reacted with the racemic anti 5,9-dimethylchrysene dihydrodiol epoxide (5,9-DMCDE). The deoxyribonucleoside adducts were separated by HPLC and characterized by CD and NMR. Approximately 17% of the epoxide was trapped by native DNA and 76% of the adducts were derived from the RSSR enantiomer. The ratios of dAdo/dGuo modification in DNA were 14/86 and 19/81 for RSSR and SRRS enantiomers, respectively. By monitoring the product yields of anti 5,9-DMCDE with DNA and deoxyribonucleotides, we hoped to gain further insight into the factors responsible for deoxyguanosine adduct formation by 5-methylchrysene dihydrodiol epoxide (5-MCDE) compared to 5,6-dimethylchrysene dihydrodiol epoxide (5,6-DMCDE). The adduct yields in deoxyribonucleotide reactions of 5,9-DMCDE were slightly higher than those from 5-MCDE. However, the reaction yields of 5,9-DMCDE with DNA were lower than those with 5-MCDE in most cases, particularly for the cis and trans deoxyadenosine adducts. It seems that the 9-methyl group of 5,9-DMCDE significantly influences adduct formation with the deoxyadenosine residue in DNA in contrast to the 6-methyl group of 5,6-DMCDE. The 9-methyl group sterically decreases deoxyadenosine adduct yields more in reaction with native DNA than denatured DNA, but it has little effect on deoxyribonucleotide reactions. Adduct formation with deoxyguanosine residues in DNA by all three dihydrodiol epoxides correlate with their respective tumorigenic and mutagenic activities. (C) 2128 Elsevier Science Ireland Ltd. All rights reserved. C1 NCI, Chem Carcinogenesis Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Amer Hlth Fdn, Naylor Dana Inst Dis Prevent, Valhalla, NY 10595 USA. RP Szeliga, J (reprint author), NCI, Chem Carcinogenesis Lab, Frederick Canc Res & Dev Ctr, Bldg 538,Room 114,POB B, Frederick, MD 21702 USA. FU NCI NIH HHS [CA 17613] NR 28 TC 4 Z9 4 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0009-2797 J9 CHEM-BIOL INTERACT JI Chem.-Biol. Interact. PD NOV 1 PY 2000 VL 128 IS 3 BP 159 EP 172 DI 10.1016/S0009-2797(00)00184-8 PG 14 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 371PP UT WOS:000165187600001 PM 11064001 ER PT J AU Kunos, G Jarai, Z Batkai, S Goparaju, SK Ishac, EJN Liu, J Wang, L Wagner, JA AF Kunos, G Jarai, Z Batkai, S Goparaju, SK Ishac, EJN Liu, J Wang, L Wagner, JA TI Endocannabinoids as cardiovascular modulators SO CHEMISTRY AND PHYSICS OF LIPIDS LA English DT Review DE cannabinoids; marijuana; cardiovascular effects ID CANNABINOID CB1 RECEPTOR; RAT-BRAIN; MESENTERIC VASODILATION; HYPERPOLARIZING FACTOR; SELECTIVE ANTAGONIST; NEUROBLASTOMA-CELLS; INDUCED HYPOTENSION; ENDOTHELIAL-CELLS; ADENYLATE-CYCLASE; BINDING ACTIVITY AB Cannabinoids, the bioactive constituents of the marijuana plant and their synthetic and endogenous analogs cause not only neurobehavioral, but also cardiovascular effects. The most important component of these effects is a profound decrease in blood pressure and heart rate. Although multiple lines of evidence indicate that the hypotensive and bradycardic effects of anandamide and other cannabinoids are mediated by peripherally located CB1 cannabinoid receptors, anandamide can also elicit vasodilation in certain vascular beds, which is independent of CB1 or CB2 receptors. Possible cellular mechanisms underlying these effects and the cellular sources of vasoactive anandamide are discussed. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. RP Kunos, G (reprint author), NIAAA, NIH, Bldg 31,Rm 1B58,31 Ctr Dr,MSC 2088, Bethesda, MD 20892 USA. RI Batkai, Sandor/G-3889-2010; Batkai, Sandor/H-7983-2014 FU NHLBI NIH HHS [HL49938, HL59257] NR 78 TC 88 Z9 92 U1 0 U2 2 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0009-3084 J9 CHEM PHYS LIPIDS JI Chem. Phys. Lipids PD NOV PY 2000 VL 108 IS 1-2 BP 159 EP 168 DI 10.1016/S0009-3084(00)00194-8 PG 10 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 381RN UT WOS:000165777100012 PM 11106789 ER PT J AU Lamb, ME Sternberg, KJ Esplin, PW AF Lamb, ME Sternberg, KJ Esplin, PW TI Effects of age and delay on the amount of information provided by alleged sex abuse victims in investigative interviews SO CHILD DEVELOPMENT LA English DT Article ID CHILDRENS MEMORY; EYEWITNESS TESTIMONY; RETENTION INTERVAL; TRAUMATIC INJURY; UTTERANCE TYPES; EVENT; REPETITION; WITNESSES; IMPACT; STYLE AB A total of 145 children of 4 to 5, 6 to 7 8 to 9, and 10 to 12 years of age were interviewed within 3 days, 1 month, 1 to 3 months, or 5 to 14 months after allegedly experiencing a single incident of sexual abuse. The proportion of substantive investigative utterances eliciting new details from the children increased with age and decreased after delays of more than 1 month. Age (but not delay) was also associated with the length and richness of informative responses to individual investigative utterances of all types. Children were more likely to provide new details in response to option-posing and suggestive prompts. As in previous field studies, interviewers employed few open-ended prompts, and thus only 5% of the information obtained was elicited using free-recall prompts. C1 NICHHD, Sect Social & Emot Dev, Bethesda, MD 20892 USA. RP Lamb, ME (reprint author), NICHHD, Sect Social & Emot Dev, 9190 Rockville Pike, Bethesda, MD 20892 USA. EM Michael_Lamb@nih.gov NR 51 TC 55 Z9 56 U1 2 U2 8 PU WILEY-BLACKWELL PI MALDEN PA COMMERCE PLACE, 350 MAIN ST, MALDEN 02148, MA USA SN 0009-3920 J9 CHILD DEV JI Child Dev. PD NOV-DEC PY 2000 VL 71 IS 6 BP 1586 EP 1596 DI 10.1111/1467-8624.00250 PG 11 WC Psychology, Educational; Psychology, Developmental SC Psychology GA 387AJ UT WOS:000166097700010 PM 11194258 ER PT J AU Picco, P Gattorno, M Marchese, N Vignola, S Sormani, MP Barabino, A Buoncompagni, A AF Picco, P Gattorno, M Marchese, N Vignola, S Sormani, MP Barabino, A Buoncompagni, A TI Increased gut permeability in juvenile chronic arthritides. A multivariate analysis of the diagnostic parameters SO CLINICAL AND EXPERIMENTAL RHEUMATOLOGY LA English DT Article DE early onset-pauciarticular juvenile chronic arthritis; late onset-pauciarticular juvenile chronic arthritis; juvenile spondyloarthropathy; intestinal permeability; lactulose/mannitol ratio ID INDUCED GASTRODUODENAL INJURY; INTESTINAL PERMEABILITY; ANKYLOSING-SPONDYLITIS; RHEUMATOID-ARTHRITIS; CHILDREN; SPONDYLOARTHROPATHIES; INFLAMMATION; EVOLUTION; DISEASES; POLYMERS AB Objective This study was aimed at evaluating intestinal permeability (IP) in patients with oligoarticular juvenile idiopathic arthritis (o-JIA) spondyloarthropathy (SpA) associated with inflammatory bowel disease (IBD) and other forms of juvenile-onset chronic arthritidis (OIA) using the lactulose/mannitol (L/M) test in comparison with other noninvasive parameters of gut involvement. Methods A series of 26 children affected with o-JIA and 14 with either SpA/IBD or OIA were assessed for IP. The urinary L/M ratio was measured by gas chromatography. The erythrocyte sedimentation rate (ESR), C reactive protein (CRP), and faecal alpha 1antitrypsin concentrations were also evaluated. Ten o-JIA patients displayed active arthritis while in 16 the disease was under control. Among the OIA patients, II were affected with psoriatic arthritis and the remaining 3 with chronic reactive arthritis. 14 patients with SpA-IBD had active synovitis or spine inflammation. 14 eo-pJCA and 22 OIA and SpA-IBD patients, respectively, were receiving NSAID therapy. Results The mean L/M ratios for the Spa-IBD (0.07 +/- 0.02, mean +/- SD), OIA (0.05 0.02) and o-JIA (0.04 +/- 0.02) patients were significantly higher (p < 0.001, p = 0.022 and p = 0.01, respectively) than those found-in controls (0.02 +/- 0.01). Logistic regression analysis disclosed a positive correlation between the L/M ratio and the presence of gastrointestinal manifestations (p = 0.011). The type of disease (p = 0.28), the disease activity in the JCA patient group (p = 0.24) and NSAID administration (p = 0.210) did not seem to significantly influence the L/M ratio. Conclusions All of the subtypes of juvenile chronic arthritides that we studied displayed an increased IP Hence, gut wall inflammation (albeit asymptomatic) may also be present in o-JIA patients. The SpA-IBD patients with gastrointestinal symptoms displayed the highest mean L/M ratio values. The L/M test seemed to correlate with histopathological features of the gut mucosa. The L/M ratio was shown to be a highly sensitive but poorly specific test for predicting gut inflammatory disease compared to other non-invasive screening tests. C1 G Gaslini Sci Inst, Div Pediat 2, I-16147 Genoa, Italy. G Gaslini Sci Inst, Biochem Lab, I-16147 Genoa, Italy. G Gaslini Sci Inst, Div Pediat 3, I-16147 Genoa, Italy. Natl Canc Inst, Clin Epidemiol Unit, Genoa, Italy. RP Picco, P (reprint author), G Gaslini Sci Inst, Div Pediat 2, Largo G Gaslini 5, I-16147 Genoa, Italy. NR 27 TC 29 Z9 30 U1 0 U2 2 PU CLINICAL & EXPER RHEUMATOLOGY PI PISA PA VIA SANTA MARIA 31, 56126 PISA, ITALY SN 0392-856X J9 CLIN EXP RHEUMATOL JI Clin. Exp. Rheumatol. PD NOV-DEC PY 2000 VL 18 IS 6 BP 773 EP 778 PG 6 WC Rheumatology SC Rheumatology GA 377TX UT WOS:000165543300024 PM 11138347 ER PT J AU Vaishampayan, U Glode, M Du, W Kraft, A Hudes, G Wright, J Hussain, M AF Vaishampayan, U Glode, M Du, W Kraft, A Hudes, G Wright, J Hussain, M TI Phase II study of dolastatin-10 in patients with hormone-refractory metastatic prostate adenocarcinoma SO CLINICAL CANCER RESEARCH LA English DT Article; Proceedings Paper CT 91st Annual Meeting of the American-Association-for-Cancer-Research CY APR 01-05, 2000 CL SAN FRANCISCO, CALIFORNIA SP Amer Assoc Canc Res ID ORAL ESTRAMUSTINE PHOSPHATE; ANTINEOPLASTIC AGENTS; ONCOLOGY-GROUP; AURISTATIN-PE; CANCER CELLS; IN-VITRO; TRIAL; MITOXANTRONE; VINBLASTINE; APOPTOSIS AB Dolastatin-10 is a natural, cytotoxic peptide with microtubule-inhibitory and apoptotic effects. It has demonstrated in vitro and in vivo efficacy in the DU-145 human prostate cancer model. A Phase II clinical trial was designed in patients with hormone-refractory prostate cancer. Dolastatin-10 was administered at a dose of 400 mug/m(2) i.v. every 3 weeks. Dose escalation to 450 mug/m(2) was permitted. Toxicity evaluation was conducted every 2 weeks, and assessment of response was done at the end of every two cycles. Sixteen patients were enrolled between October 1998 to December 1999. The median age was 71 Sears (range, 59-79 years). Median prostate-specific antigen value was 108 ng/ml (range, 15.3-1672 ng/ml). Of the 15 eligible patients, 7 were Caucasian and 8 were African-American. Eight patients had bone-only metastases, and seven had measurable disease with or without bone metastases. A total of 56 cycles have been administered. Only 2 patients required dose adjustment because of toxicity, and in 5 patients, dose escalation was feasible to 450 mug/m(2). The major toxicities observed were grade 3 and 4 neutropenia in 8 patients and grade 3 neuropathy in 1 patient. All 15 patients are evaluable for response. Three patients demonstrated stable disease; 2 of these had bone disease, and 1 had nodal metastasis. All others had disease progression. Dolastatin-10 is very well tolerated in this elderly, pretreated population hut lacks significant clinical activity as a single agent. C1 Wayne State Univ, Dept Biostat, Detroit, MI 48201 USA. Wayne State Univ, Dept Med, Div Hematol Oncol, Detroit, MI 48201 USA. Barbara Ann Karmanos Canc Inst, Detroit, MI 48201 USA. Fox Chase Canc Ctr, Philadelphia, PA 19111 USA. NCI, Bethesda, MD 20892 USA. RP Hussain, M (reprint author), Harper Grace Hosp, 5 Hudson,3990 John R Rd, Detroit, MI 48201 USA. NR 32 TC 89 Z9 97 U1 1 U2 6 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 IS 11 BP 4205 EP 4208 PG 4 WC Oncology SC Oncology GA 374LX UT WOS:000165347600006 PM 11106233 ER PT J AU Holten-Andersen, MN Stephens, RW Nielsen, HJ Murphy, G Christensen, IJ Stetler-Stevenson, W Brunner, N AF Holten-Andersen, MN Stephens, RW Nielsen, HJ Murphy, G Christensen, IJ Stetler-Stevenson, W Brunner, N TI High preoperative plasma tissue inhibitor of metalloproteinase-1 levels are associated with short survival of patients with colorectal cancer SO CLINICAL CANCER RESEARCH LA English DT Article ID UROKINASE-PLASMINOGEN-ACTIVATOR; HUMAN COLON CANCER; BREAST-CANCER; MATRIX METALLOPROTEINASES; DISTANT METASTASES; PROGNOSIS; COLLAGENASE; CELLS; RECEPTOR; TUMOR AB The objective of the present study was to measure preoperative plasma tissue inhibitor of metalloproteinase (TIMP)-1 levels in colorectal cancer patients and relate these values to clinical and biochemical patient characteristics. TIMP-1 levels were determined by ELISA in EDTA plasma samples collected preoperatively from 588 colorectal cancer patients. Plasma TIMP-1 levels were distributed with a median value of 141.1 mug/liter (range, 53.7-788.7 mug/liter). Whereas no significant differences were found in the median plasma TIMP-1 levels among patients with Dukes' stage A, B, and C disease, patients with Dukes' stage D disease had significantly higher plasma TIMP-1 levels (P < 0.0001); however, high plasma TIMP-1 levels were not restricted to advanced disease. A relatively weak correlation between plasma TIMP-1 level and age was found (r = 0.35; P < 0.0001), There was no significant difference in TIMP-1 levels between males and females (P = 0.97), Univariate analysis demonstrated an increasing risk of mortality with increasing TIMP-1 levels [scored as the log,(TIMP-1); hazard ratio 33; 95% confidence interval, 2.6-4.2; P < 0.0001], Including covariates (Dukes' stage, primary tumor location, gender, age, plasminogen activator inhibitor type 1, and soluble urokinase plasminogen activator receptor) in a multivariate analysis, TIMP-1 was retained in the final model (hazard ratio = 2.5; 95% confidence interval, 1.7-3.7; P < 0.0001), This study showed a highly significant association between preoperative plasma TIMP-1 levels and survival in colorectal canter patients, with higher plasma TIMP-1 levels being associated with poor outcome, independent of clinical parameters including Dukes' stage, plasma TIMP-1 levels were found to strongly predict prognosis of colorectal ranter patients. Additional studies are needed to validate the clinical usefulness of plasma TIMP-1 measurements. C1 Rigshosp, Finsen Lab, DK-2100 Copenhagen, Denmark. Hvidovre Univ Hosp, Dept Surg Gastroenterol, DK-2650 Hvidovre, Denmark. Univ E Anglia, Sch Biol Sci, Norwich NR4 7T3, Norfolk, England. NCI, Pathol Lab, Bethesda, MD 20892 USA. RP Brunner, N (reprint author), Finsen Lab, Stradblvd 49, DK-2100 Copenhagen, Denmark. RI Stephens, Ross/A-8524-2008; Stetler-Stevenson, William/H-6956-2012 OI Stephens, Ross/0000-0001-6124-0102; Stetler-Stevenson, William/0000-0002-5500-5808 NR 48 TC 125 Z9 130 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 IS 11 BP 4292 EP 4299 PG 8 WC Oncology SC Oncology GA 374LX UT WOS:000165347600019 PM 11106246 ER PT J AU Arbuck, SG Pluda, J Korn, E Hunsberger, S Christian, MD Grochow, L Rubinstein, L AF Arbuck, SG Pluda, J Korn, E Hunsberger, S Christian, MD Grochow, L Rubinstein, L TI Incorporating biological endpoints into phase 1 trials: New designs. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20854 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 003 BP 4472S EP 4472S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000005 ER PT J AU Green, JE AF Green, JE TI Validation of genetically-engineered animal models for cancer and their use in pre-clinical testing. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 001 BP 4472S EP 4472S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000003 ER PT J AU Rybak, SM AF Rybak, SM TI Novel RNA damaging agents for cancer therapy. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Frederick Canc Res & Dev Ctr, Dev Therapeut Program, Div Canc Treatment & Diag, Frederick, MD 21702 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 011 BP 4475S EP 4475S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000013 ER PT J AU Holbeck, S Zaharevitz, D Sausville, E AF Holbeck, S Zaharevitz, D Sausville, E TI The NCI 60 human tumor cell line panel: A tool linking molecular targets to drug sensitivity. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Dev Therapeut Program, Rockville, MD 20852 USA. NR 0 TC 0 Z9 0 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 016 BP 4476S EP 4476S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000018 ER PT J AU Pommier, Y AF Pommier, Y TI Topoisomerase I-mediated DNA damage and repair. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 030 BP 4479S EP 4479S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000032 ER PT J AU Ford, LG Dunn, BK AF Ford, LG Dunn, BK TI SERMS in action: Ongoing research in breast cancer prevention. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 034 BP 4480S EP 4480S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000036 ER PT J AU Senderowicz, AM AF Senderowicz, AM TI Preclinical and clinical development of CDK inhibitors. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NIH, Mol Therapeut Unit, Oral Pharyngeal Canc Branch, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 042 BP 4482S EP 4482S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000044 ER PT J AU Llebes, L Hochster, H Conaway, CC Chachoua, A Farrell, K Lee, J Mendoza, S Crowell, J Carmella, C Murphy, S Hecht, SS Chung, F AF Llebes, L Hochster, H Conaway, CC Chachoua, A Farrell, K Lee, J Mendoza, S Crowell, J Carmella, C Murphy, S Hecht, SS Chung, F TI Thirty day multiple dose administration of PEITC: A phase I study with steady state pharmacokinetics. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NYU, Sch Med, New York, NY USA. NCI, Am Hlth Fdn, Bethesda, MD 20892 USA. NCI, Chemoprevent Branch, Bethesda, MD 20892 USA. Univ Minnesota, Ctr Canc, Minneapolis, MN 55455 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 106 BP 4486S EP 4487S PG 2 WC Oncology SC Oncology GA 375NW UT WOS:000165409000066 ER PT J AU Cushman, M Jayaraman, M Vroman, JA Fukunaga, AK Fox, B Kohlhagen, G Strumberg, D Pommier, Y AF Cushman, M Jayaraman, M Vroman, JA Fukunaga, AK Fox, B Kohlhagen, G Strumberg, D Pommier, Y TI Synthesis of new indeno[1, 2-c]isoquinolines: Cytotoxic non-camp-tothecin topoisomerase I inhibitors. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Purdue Univ, Sch Pharm, W Lafayette, IN 47907 USA. NCI, DBS, LMP, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 4 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 135 BP 4493S EP 4493S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000095 ER PT J AU Rabow, AA Covell, DG Sausville, EA Shoemaker, RH AF Rabow, AA Covell, DG Sausville, EA Shoemaker, RH TI Development of novel computational tools for drug discovery. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Lab Computat Biol, NIH, Bethesda, MD 20892 USA. NCI, DCT, Dev Therapeut Program, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 154 BP 4497S EP 4497S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000114 ER PT J AU Potapova, O Anisimov, SV Boheler, K Holbrook, NJ AF Potapova, O Anisimov, SV Boheler, K Holbrook, NJ TI A role of JNK signaling in human prostate cancer: Results of the SAGE analysis. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NIA, NIH, Baltimore, MD 21224 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 168 BP 4500S EP 4500S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000128 ER PT J AU Rapisarda, A Uranchimeg, B Scudiero, DA Selby, MH Stevenson, TJ Sausville, EA Shoemaker, RH Melillo, G AF Rapisarda, A Uranchimeg, B Scudiero, DA Selby, MH Stevenson, TJ Sausville, EA Shoemaker, RH Melillo, G TI Hypoxia inducible factor 1: A molecular target for development of novel cancer therapeutics. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 SAIC Frederick, Frederick, MD USA. NCI, Frederick Canc Res & Dev Ctr, Dev Therapeut Program, Screening Technol Branch, Frederick, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 171 BP 4500S EP 4500S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000131 ER PT J AU Reinhold, WC Kohn, KW Urasaki, Y Pantazis, P Miller, L Liu, E Pommier, YG Weinstein, JN AF Reinhold, WC Kohn, KW Urasaki, Y Pantazis, P Miller, L Liu, E Pommier, YG Weinstein, JN TI Altered expression levels of genes involved in the apoptotic and nucleotide excision repair pathways in camptothecin-resistant cells: A cDNA microarray analysis. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, DBS, LMP, NIH, Bethesda, MD 20892 USA. NCI, ATC, Gaithersburg, MD USA. Brown Univ, Providence, RI 02912 USA. RI Liu, Edison/C-4141-2008 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 193 BP 4504S EP 4504S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000153 ER PT J AU Takebayashi, Y Zimonjic, DB Fukumoto, M Pourquier, P Emmert, S Urasaki, Y Nakayama, K Kraemer, KH Popescu, N Pommier, Y AF Takebayashi, Y Zimonjic, DB Fukumoto, M Pourquier, P Emmert, S Urasaki, Y Nakayama, K Kraemer, KH Popescu, N Pommier, Y TI Nucleotide excision repair-dependent cytotoxicity of Ecteinascidin 743. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, LMP, NIH, Bethesda, MD 20892 USA. NCI, LEC, NIH, Bethesda, MD 20892 USA. NCI, BRL, NIH, Bethesda, MD 20892 USA. NCI, DBS, NIH, Bethesda, MD 20892 USA. Tohoku Univ, Sendai, Miyagi 980, Japan. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 207 BP 4508S EP 4508S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000166 ER PT J AU Burke, TG Houghton, PJ Born, D Yang, D Wu, D Kruszewski, S Pommier, Y Kohlhagen, G Perez-Soler, R Zimmer, SG Strode, JT Chavan, AJ Latus, LJ Curran, DP AF Burke, TG Houghton, PJ Born, D Yang, D Wu, D Kruszewski, S Pommier, Y Kohlhagen, G Perez-Soler, R Zimmer, SG Strode, JT Chavan, AJ Latus, LJ Curran, DP TI NCl raid-sponsored DB-67 displays elevated lactone levels in human blood and potent anticancer activity in vitro and in vivo. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Kentucky, Lexington, KY 40506 USA. St Jude Childrens Res Hosp, Memphis, TN 38105 USA. NCI, Bethesda, MD 20892 USA. NYU, Sch Med, New York, NY USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. RI Kruszewski, Stefan/H-3265-2014 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 225 BP 4511S EP 4511S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000184 ER PT J AU Gamcsik, MP Colvin, OM Kohlhagen, G Urasaki, Y Kasibhatia, MS Adams, DJ Flowers, JL Pommier, Y Manikumar, G Wani, MC Wall, ME AF Gamcsik, MP Colvin, OM Kohlhagen, G Urasaki, Y Kasibhatia, MS Adams, DJ Flowers, JL Pommier, Y Manikumar, G Wani, MC Wall, ME TI Glutathione conjugate of camptothecin is cytotoxic and enhances the stability of the topoisomerase I-DNA cleavage complex. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Duke Univ, Durham, NC USA. NCI, Bethesda, MD 20892 USA. Res Triangle Inst, RTP, NC USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 224 BP 4511S EP 4511S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000183 ER PT J AU Yang, D Kruszewski, S Bom, D Zimmer, SG Pommier, Y Kohlhagen, G Bingcang, AL Du, W Curran, DP Burke, TG AF Yang, D Kruszewski, S Bom, D Zimmer, SG Pommier, Y Kohlhagen, G Bingcang, AL Du, W Curran, DP Burke, TG TI The 10-hydroxy functionality markedly impact on the physicochemical and pharmacological properties of the novel, highly lipophilic camptothecin analogs. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Kentucky, Lexington, KY 40506 USA. NCI, Bethesda, MD 20892 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. RI Yang, Danzhou/B-6228-2012; Kruszewski, Stefan/H-3265-2014 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 226 BP 4511S EP 4511S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000185 ER PT J AU Hamilton, M Sparreboom, A Loos, W Verweij, J Demetri, G Gelmon, K Eisenhauer, E Warren, S Ptaszynski, M AF Hamilton, M Sparreboom, A Loos, W Verweij, J Demetri, G Gelmon, K Eisenhauer, E Warren, S Ptaszynski, M TI Pharmacokinetics of NX 211 (liposomal lurtotecan) in phase I patients with solid tumors. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Gilead Sci Inc, Boulder, CO USA. Rotterdam Canc Inst, Rotterdam, Netherlands. Dana Farber Partners Canc Care, Boston, MA USA. British Columbia Canc Agcy, Vancouver, BC V5Z 4E6, Canada. NCI, Canada Clin Trials Grp, Kingston, ON, Canada. RI Sparreboom, Alex/B-3247-2008 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 238 BP 4513S EP 4514S PG 2 WC Oncology SC Oncology GA 375NW UT WOS:000165409000197 ER PT J AU Herbst, R Tran, H Hess, K Madden, T Charnsangavej, C Gravel, D Terry, K Guerra, M Taebel, K Ellis, L Pluda, J Hong, WK Abbruzzese, J AF Herbst, R Tran, H Hess, K Madden, T Charnsangavej, C Gravel, D Terry, K Guerra, M Taebel, K Ellis, L Pluda, J Hong, WK Abbruzzese, J TI A phase I clinical trial of recombinant human endostatin (rHE) in patients (PTS) with solid tumors: Pharmacokinetic (PK), safety and efficacy analysis. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. NCI, CTEP, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 259 BP 4518S EP 4518S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000218 ER PT J AU Figg, WD Dahut, W Libutti, SK Carrasquillo, J Bacharach, S Kurdziel, K Huebsch, F Kruger, EA Pluda, J Reed, E AF Figg, WD Dahut, W Libutti, SK Carrasquillo, J Bacharach, S Kurdziel, K Huebsch, F Kruger, EA Pluda, J Reed, E TI Thalidomide, an angiogenesis inhibitor, has activity in metastatic prostate cancer. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. RI Carrasquillo, Jorge/E-7120-2010; Figg Sr, William/M-2411-2016 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 285 BP 4522S EP 4523S PG 2 WC Oncology SC Oncology GA 375NW UT WOS:000165409000244 ER PT J AU Rudek, MA Dyer, V Dahut, W Pluda, JM Reed, E Figg, WD AF Rudek, MA Dyer, V Dahut, W Pluda, JM Reed, E Figg, WD TI Pharmacokinetics (PK) of COL-3, a matrix metalloproteinase (MMP) inhibitor. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Med Branch, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 293 BP 4524S EP 4524S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000251 ER PT J AU Zhai, S Figg, WD Headlee, D Messmann, RA Arbuck, SG Murgo, A Melillo, G Sausville, EA Senderowicz, AM AF Zhai, S Figg, WD Headlee, D Messmann, RA Arbuck, SG Murgo, A Melillo, G Sausville, EA Senderowicz, AM TI Pharmacokinetics (PK) of flavopiridol IV bolus in patients with refractory neoplasms. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Med Branch, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 0 TC 2 Z9 2 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 314 BP 4529S EP 4529S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000272 ER PT J AU Cholody, WM Kosakowska-Cholody, T Hollingshead, M Michejda, CJ AF Cholody, WM Kosakowska-Cholody, T Hollingshead, M Michejda, CJ TI Abrogators of checkpoint controls induce apoptosis in colon cancer cells that are growth arrested by the bisimidazoacridone, WMC26. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Frederick Canc Res & Dev Ctr, Mol Aspects Drug Design Sect, SBL,DBS, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Dev Therapeut Program, Frederick, MD 21702 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 320 BP 4530S EP 4530S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000278 ER PT J AU Dancey, J Price, W Arbuck, S Grochow, L AF Dancey, J Price, W Arbuck, S Grochow, L TI Relationship of platelet (plt) nadir to bleeding in patients (pts) on early clinical trials: Analysis of NCI clinical trials monitoring system (CTMS) database. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Canc Therapy Evaluat Program, Rockville, MD USA. Theradex, Princeton, NJ USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 351 BP 4536S EP 4536S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000309 ER PT J AU Soga, S Shiotsu, Y Sharma, SV Ikuina, Y Murakata, C Tamaoki, T Kurebayasi, J Schulte, TW Neckers, LM Akinaga, S AF Soga, S Shiotsu, Y Sharma, SV Ikuina, Y Murakata, C Tamaoki, T Kurebayasi, J Schulte, TW Neckers, LM Akinaga, S TI Against erbB-2 over-expressing breast cancer cells or chronic myelogenous leukemia cells with bcr-abl translocation thorough destabilization of erbB-2 or bcr-abl oncoproteins. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Kawasaki Med Sch, Kawasaki, Kanagawa, Japan. NCI, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 399 BP 4546S EP 4546S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000357 ER PT J AU Hamilton, A Eder, JP Pavlick, A Clark, JW Chachoua, A Ryan, DP Farrell, K Wasserstrom, H Liebes, L Wright, J Elliott, P Adams, J Muggia, F AF Hamilton, A Eder, JP Pavlick, A Clark, JW Chachoua, A Ryan, DP Farrell, K Wasserstrom, H Liebes, L Wright, J Elliott, P Adams, J Muggia, F TI Proteasome inhibition by PS-341: A phase I study. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NYU, Sch Med, New York, NY USA. Dana Farber Canc Inst, Boston, MA 02115 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. NCI, CTEP, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 417 BP 4549S EP 4549S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000375 ER PT J AU Millard, JW El-sayed, MM Yalkowsky, SH Tabibi, SE AF Millard, JW El-sayed, MM Yalkowsky, SH Tabibi, SE TI Effect of pH control on the solubilization of NSC-674495 by solubilizing agents. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Arizona, Coll Pharm, Tucson, AZ 85721 USA. NCI, DCTD, DTP, PRB, Rockville, MD 20852 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 447 BP 4556S EP 4556S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000405 ER PT J AU Zhao, LW Ran, YQ Tabibi, E Yalkowsky, SH AF Zhao, LW Ran, YQ Tabibi, E Yalkowsky, SH TI Formulation and stability evaluation of stealth liposomes containing the antitumor agent 2-(4 '-amino-3 '-methylphenyl)-benzothiazole, NSC674495, (AMPB). SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Arizona, Coll Pharm, Tucson, AZ 85721 USA. NCI, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 448 BP 4556S EP 4556S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000406 ER PT J AU Demir, AS Tanyeli, C Zimmer, S Kruszewski, S Bom, D Yang, D Chavan, A Pommier, Y Burke, TG AF Demir, AS Tanyeli, C Zimmer, S Kruszewski, S Bom, D Yang, D Chavan, A Pommier, Y Burke, TG TI Development and evaluation of a remote-loadable liposomal camptothecin which contains a polyamine functionality at position 7. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Kentucky, Lexington, KY 40506 USA. NCI, Bethesda, MD 20892 USA. RI Kruszewski, Stefan/H-3265-2014 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 453 BP 4557S EP 4557S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000411 ER PT J AU Newton, DL Mikulski, SM Sausville, EA Rybak, SM AF Newton, DL Mikulski, SM Sausville, EA Rybak, SM TI Targeting the CD22 receptor on human B-cell lymphomas with RNA damaging agents. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 NCI, Frederick Canc Res & Dev Ctr, DCTD, DTP, Frederick, MD 21702 USA. Alfacell Corp, Bloomfield, NJ 07003 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 469 BP 4560S EP 4560S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000427 ER PT J AU Mockel, B Burger, A Sausville, E Schultz, RJ Wilhelm-Ogunbiyi, K Langer, M Eck, J Zinke, H Fiebig, HH Lentzen, H AF Mockel, B Burger, A Sausville, E Schultz, RJ Wilhelm-Ogunbiyi, K Langer, M Eck, J Zinke, H Fiebig, HH Lentzen, H TI Antitumour activity of rViscumin in vitro and in vivo. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 BRAIN GmbH, Zwingenberg, 20892, Germany. Univ Freiburg, Freiburg, Germany. NCI, Bethesda, MD USA. MADAUS AG, Cologne, Germany. VISCUM AG, Zwingenberg, Germany. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA 615 CHESTNUT ST, 17TH FLOOR, PHILADELPHIA, PA 19106-4404 USA SN 1078-0432 EI 1557-3265 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 490 BP 4564S EP 4565S PG 2 WC Oncology SC Oncology GA 375NW UT WOS:000165409000448 ER PT J AU Lans, TE Libutti, S Alexander, HR Wu, P ten Hagen, TLM Eggermont, AMM AF Lans, TE Libutti, S Alexander, HR Wu, P ten Hagen, TLM Eggermont, AMM TI Induction of tumor sensitivity to TNF by means of transfection of tumor with an Endothelial Monocyte Activating Polypeptide (EMAP II) gene. SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 AZR Daniel den Hoed, Dept Surg Oncol, Bethesda, MD USA. NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 512 BP 4569S EP 4569S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000470 ER PT J AU Herbst, R Hess, K Mulliani, N Charnsangavej, C Baker, C Ellis, L Kim, E Bucana, C Pluda, J Fidler, I Abbruzzese, J AF Herbst, R Hess, K Mulliani, N Charnsangavej, C Baker, C Ellis, L Kim, E Bucana, C Pluda, J Fidler, I Abbruzzese, J TI A phase I clinical trial of recombinant human endostatin (rHE) in patients (PTS) with solid tumors: Surrogate analyses to determine a biologically effective dose (BED). SO CLINICAL CANCER RESEARCH LA English DT Meeting Abstract C1 Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. NCI, CTEP, NIH, Bethesda, MD 20892 USA. NR 0 TC 3 Z9 3 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD NOV PY 2000 VL 6 SU S MA 578 BP 4582S EP 4582S PG 1 WC Oncology SC Oncology GA 375NW UT WOS:000165409000534 ER PT J AU Ellahham, SH Charlon, V Abassi, Z Calis, KA Choucair, WK AF Ellahham, SH Charlon, V Abassi, Z Calis, KA Choucair, WK TI Bosentan and the endothelin system in congestive heart failure SO CLINICAL CARDIOLOGY LA English DT Review DE endothelin; bosentan; chronic heart failure; receptor antagonist ID SPONTANEOUSLY HYPERTENSIVE RATS; RECEPTOR ANTAGONIST BOSENTAN; PLASMA ENDOTHELIN; RELAXING FACTOR; ET(B) RECEPTOR; NITRIC-OXIDE; VASOCONSTRICTION; CONTRACTION; EXPRESSION; KIDNEY AB The endothelin system appears to play an important role in the pathophysiology of congestive heart failure (CHF). Endothelin receptor antagonists represent a novel class of agents that are being evaluated for their potential benefits in treating various cardiovascular disorders. Bosentan is an orally active endothelin receptor antagonist that has been studied for the treatment of CHE Early clinical experience with bosentan has confirmed some benefits on hemodynamic parameters in patients with CHE Its role in slowing the progression of the disease and improving survival remains to be elucidated. C1 Washington Hosp Ctr, Div Cardiol, Washington, DC 20010 USA. Hoffmann La Roche, Basel, Switzerland. NIH, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Ellahham, SH (reprint author), Washington Hosp Ctr, Div Cardiol, Room 1103,110 Irving St NW, Washington, DC 20010 USA. NR 60 TC 12 Z9 14 U1 1 U2 1 PU CLINICAL CARDIOLOGY PUBL CO PI MAHWAH PA PO BOX 832, MAHWAH, NJ 07430-0832 USA SN 0160-9289 J9 CLIN CARDIOL JI Clin. Cardiol. PD NOV PY 2000 VL 23 IS 11 BP 803 EP 807 PG 5 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 371QM UT WOS:000165189700004 PM 11097125 ER PT J AU Khripin, Y Myakishev, MV Hu, S Lueders, K Hamer, D AF Khripin, Y Myakishev, MV Hu, S Lueders, K Hamer, D TI A new method for SNP genotyping based on allele-specific PCR with universal FRET primers. SO CLINICAL CHEMISTRY LA English DT Meeting Abstract C1 Intergen, Gaithersburg, MD USA. NCI, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD NOV PY 2000 VL 46 IS 11 MA 18 BP 1877 EP 1877 PG 1 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 372HR UT WOS:000165228700057 ER PT J AU Slavotinek, AM Vacha, SJ Peters, KF Biesecker, LG AF Slavotinek, AM Vacha, SJ Peters, KF Biesecker, LG TI Sudden death caused by pulmonary thromboembolism in Proteus syndrome SO CLINICAL GENETICS LA English DT Article DE Klippel-Trenaunay syndrome; Proteus syndrome; pulmonary thromboembolism ID KLIPPEL-TRENAUNAY-SYNDROME; WEBER-SYNDROME; DIFFERENTIAL-DIAGNOSIS; SOMATIC MOSAICISM; MANAGEMENT; HEMIHYPERTROPHY; MANIFESTATIONS; LIPOMATOSIS; PHENOTYPE; FEATURES AB We report 3 patients with Proteus syndrome (PS) who died suddenly from pulmonary embolism (PE). The first patient was a male diagnosed with PS at 12 years who had varicose veins, portal vein thrombosis, right iliac vein occlusion and recurrent PE. At age 25 years, he was admitted to the hospital with a severe headache. Despite therapeutic doses of warfarin, investigations for an acute episode of breathlessness showed PE and he was unable to be resuscitated. The second case was a 9-year-old male with PS who collapsed at home and could not be revived. Autopsy revealed that the cause of death was a PE associated with thrombosis of the deep veins (DVT). The third patient was a 17-year-old female undergoing inpatient treatment for sinusitis when she unexpectedly arrested. She could not be revived and a full autopsy revealed a large PE with no identified DVT. We conclude that PE is a serious complication of PS and recommend vigilance concerning the signs and symptoms of thrombosis and PE in individuals with PS, including children. Aggressive evaluation and treatment should be considered urgently in patients with PS and signs or symptoms of DVT. C1 NHGRI, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. NHGRI, Off Clin Director, NIH, Bethesda, MD 20892 USA. RP Biesecker, LG (reprint author), NHGRI, Genet Dis Res Branch, NIH, Room 4B75,Bldg 49,49 Convent Dr, Bethesda, MD 20892 USA. NR 39 TC 33 Z9 35 U1 0 U2 0 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0009-9163 J9 CLIN GENET JI Clin. Genet. PD NOV PY 2000 VL 58 IS 5 BP 386 EP 389 DI 10.1034/j.1399-0004.2000.580509.x PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 380LK UT WOS:000165703200008 PM 11140839 ER PT J AU Cuccherini, B Chua, K Gill, V Weir, S Wray, B Stewart, D Nelson, D Fuss, I Strober, W AF Cuccherini, B Chua, K Gill, V Weir, S Wray, B Stewart, D Nelson, D Fuss, I Strober, W TI Bacteremia and skin/bone infections in two patients with X-linked agammaglobulinemia caused by an unusual organism related to Flexispira/Helicobacter species SO CLINICAL IMMUNOLOGY LA English DT Article DE agammaglobulinemia; bacteremia; Helicobacter; immunodeficiency ID HELICOBACTER-CINAEDI BACTEREMIA; FLEXISPIRA-RAPPINI BACTEREMIA; CAMPYLOBACTER-CINAEDI; TYROSINE KINASE; DEFECTIVE GENE; HYPOGAMMAGLOBULINEMIA; IDENTIFICATION; ARTHRITIS; BLOOD; AIDS AB Two patients with Bruton's X-linked agammaglobulinemia are described with bacteremia and skin/bone infection due to an organism which by 16S rRNA gene sequence analysis was most closely related to "Flexispira" rappini land thus designated a Flexispira-like organism, FLO) and more distantly related to the Helicobacter species, The organism required microaerobic conditions and, supplemental H-2 gas for growth and was reliably stained with acridine orange. In common with Helicobacter cinaedi infections, the focus of the FLO infection was in one case in the blood vessels or lymphatics of an extremity and in the other case in the skin and adjacent bone of an extremity. In both cases, prolonged TV antibiotic therapy was necessary to clear the infection. The susceptibility of XLA patients to FLO infection appears to be related to the fact that XLA is associated with severe B cell (humoral) immunodeficiency and thus these patients have difficulty with intravascular or intralymphatic infection. These findings elucidate the nature of FLO infections in humans and point the way to their detection and treatment, (C) 2000 Academic Press. C1 NIAID, Clin Invest Lab, Mucosal Immun Sect, NIH, Bethesda, MD 20892 USA. NCI, Dept Clin Pathol, Microbiol Serv, NIH, Bethesda, MD 20892 USA. NCI, Metab Branch, Immunophysiol Sect, NIH, Bethesda, MD 20892 USA. Med Coll Georgia, Augusta, GA 30912 USA. RP Cuccherini, B (reprint author), NIAID, Clin Invest Lab, Mucosal Immun Sect, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 43 TC 30 Z9 32 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1521-6616 J9 CLIN IMMUNOL JI Clin. Immunol. PD NOV PY 2000 VL 97 IS 2 BP 121 EP 129 DI 10.1006/clim.2000.4932 PG 9 WC Immunology SC Immunology GA 375BX UT WOS:000165380600006 PM 11027452 ER PT J AU Bennett, J AF Bennett, J TI Editorial response: Choosing amphotericin B formulations - Between a reck and a hard place SO CLINICAL INFECTIOUS DISEASES LA English DT Editorial Material ID NEUTROPENIA; FEVER C1 NIAID, Clin Invest Lab, Clin Mycol Sect, NIH, Bethesda, MD 20854 USA. RP Bennett, J (reprint author), NIAID, Clin Invest Lab, Clin Mycol Sect, NIH, Bldg 10, Bethesda, MD 20854 USA. NR 3 TC 10 Z9 10 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD NOV PY 2000 VL 31 IS 5 BP 1164 EP 1165 DI 10.1086/317443 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 380LY UT WOS:000165704400011 PM 11073746 ER PT J AU Hospenthal, DR Bennett, JE AF Hospenthal, DR Bennett, JE TI Persistence of cryptococcomas on neuroimaging SO CLINICAL INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 14th International Society-of-Human-and-Animal-Mycoses World Congress CY MAY, 2000 CL BUENOS AIRES, ARGENTINA SP Soc Human Anim Mycoses ID CENTRAL-NERVOUS-SYSTEM; CNS CRYPTOCOCCOSIS; INTRACRANIAL CRYPTOCOCCOSIS; MR FINDINGS; PATHOLOGICAL CORRELATION; MASS LESIONS; CT; AIDS; MENINGOENCEPHALITIS; INVOLVEMENT AB Three previously normal patients with cryptococcal meningitis had intracranial lesions on computed tomography and magnetic resonance imaging that persisted for >5 years after successful cure with antifungal drugs. Persistence of lesions on neuroimaging should not be misinterpreted as evidence of active cryptococcosis. C1 NIAID, Clin Mycol Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Hospenthal, DR (reprint author), Tripler Army Med Ctr, Infect Dis Serv, MCHK DMI, Honolulu, HI 96859 USA. NR 24 TC 24 Z9 24 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD NOV PY 2000 VL 31 IS 5 BP 1303 EP 1306 DI 10.1086/317434 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 380LY UT WOS:000165704400037 PM 11073772 ER PT J AU Owiny, JR Strandberg, JD AF Owiny, JR Strandberg, JD TI Cholesterol granulomas in mice deficient in apolipoprotein E SO CONTEMPORARY TOPICS IN LABORATORY ANIMAL SCIENCE LA English DT Article ID LESIONS AB Three 12- to 16-month-old female B6,129 apolipoprotein E-deficient mice were presented for necropsy because of bilateral masses in the gluteal region. Histopathologic examination revealed cholesterol granulomas extending from the superficial dermis to the underlying gluteal muscles. Microscopic granulomas were found on the face, perivaginal tissues, mesenteric lymph nodes, spleen, joints, kidneys, and choroid plexus. Other lesions included severe atherosclerosis of the aortic valves, aorta, pulmonary artery, and renal artery. Here we show that macroscopic cholesterol granulomas can develop in apolipoprotein E-deficient that receive a normal rodent diet. C1 Johns Hopkins Univ, Sch Med, Baltimore, MD 21205 USA. NIH, NCRR, Bethesda, MD 20892 USA. RP Owiny, JR (reprint author), Johns Hopkins Univ, Sch Med, 459 Ross Res Bldg,720 Rutland Ave, Baltimore, MD 21205 USA. FU NCRR NIH HHS [T32 RR07002] NR 9 TC 3 Z9 3 U1 0 U2 0 PU AMER ASSOC LABORATORY ANIMAL SCIENCE PI MEMPHIS PA 9190 CRESTWYN HILLS DR, MEMPHIS, TN 38125 USA SN 1060-0558 J9 CONTEMP TOP LAB ANIM JI Contemp. Top. Lab. Anim. Sci. PD NOV PY 2000 VL 39 IS 6 BP 57 EP 58 PG 2 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 384KW UT WOS:000165942400014 PM 11487256 ER PT J AU Sax, CM Kays, WT Salamon, C Chervenak, MM Xu, YS Piatigorsky, J AF Sax, CM Kays, WT Salamon, C Chervenak, MM Xu, YS Piatigorsky, J TI Transketolase gene expression in the cornea is influenced by environmental factors and developmentally controlled events SO CORNEA LA English DT Article DE gene expression; transketolase; light regulation; crystallin ID MAJOR SOLUBLE-PROTEIN; ALDEHYDE DEHYDROGENASE; BOVINE CORNEA; LENS CRYSTALLINS; MOUSE; EYE; IDENTIFICATION; EVOLUTION; TRANSPARENCY; EPITHELIUM AB Purpose. Transketolase (TKT) has been proposed to be a corneal crystallin, and its gene and protein are abundantly expressed in the corneal epithelium of several mammals. A marked up-regulation of TKT gene expression coincides with the time of eyelid opening in the mouse. Here, we examined whether exposure to incident light contributes to the up-regulation of TKT gene expression during cornea maturation. Methods. Mice were raised in either standard light/dark cycling conditions or total darkness. In some cases, subcutaneous injections of epidermal growth factor (EGF) were given beginning on the day of birth to induce early eyelid opening. RNA was prepared from the corneas of mothers and pups and subjected to Northern blot analyses. In addition, the relative levels of TKT mRNA and/or enzyme activity were examined in the corneas of human, bovine, rat, chicken, and zebrafish. Results. TKT mRNA levels were 2.1-fold higher in the corneas of 25-day-old mouse pups (12 days after eyelid opening) that had been born and raised in light/dark conditions compared to pups born and raised in total darkness. By contrast, the level of TKT mRNA in the mature corneas of adult mice maintained in the dark for 2-8 weeks did not vary greatly from those of mice maintained in light/dark conditions. Interestingly, TKT mRNA levels in the corneas of dark-raised mice, although reduced, did exhibit the increase characteristically observed before and after eyelid opening. In addition, TKT mRNA levels were elevated fivefold in the corneas of 28-day-old mice raised in darkness and injected with EGF compared to uninjected mice also deprived of light. The EGF-injected mice opened their eyes 3 days early, and their corneal epithelium did not grossly differ from that of control mice. TKT mRNA and/or enzyme activity was found to be much higher in the corneas than in other tissues of humans, bovines, and rats but was extremely low in the corneas of chicken and zebrafish. Conclusion, Our studies suggest that both exposure to incident light and events surrounding the process of eyelid opening play a role in the up-regulation of TKT gene expression observed during corneal maturation in mice. Light appears to play a less important role in the mature cornea in maintaining high levels of TKT gene expression. The low levels of TKT in the cornea of chicken and zebrafish support the notion that TKT acts as a taxon-specific enzyme-crystallin in mammals. The involvement of environmental signals for this putative, mammalian cornea crystallin contrasts with the purely developmental signals involved in the up-regulation of the crystallin genes of the lens. C1 NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Piatigorsky, J (reprint author), NEI, Mol & Dev Biol Lab, NIH, 6 Ctr Dr,Bldg 6 Room 201, Bethesda, MD 20892 USA. NR 37 TC 23 Z9 24 U1 1 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0277-3740 J9 CORNEA JI Cornea PD NOV PY 2000 VL 19 IS 6 BP 833 EP 841 DI 10.1097/00003226-200011000-00014 PG 9 WC Ophthalmology SC Ophthalmology GA 372JK UT WOS:000165230400013 PM 11095059 ER PT J AU Susla, G Masur, H AF Susla, G Masur, H TI Critical care pharmacists: An essential or a luxury? SO CRITICAL CARE MEDICINE LA English DT Editorial Material DE pharmacists; critical care; pharmacy services; guidelines ID UNIT; INTERVENTIONS; SERVICES; COST C1 NIH, Ctr Clin, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Susla, G (reprint author), NIH, Ctr Clin, Dept Crit Care Med, Bethesda, MD 20892 USA. NR 12 TC 2 Z9 2 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD NOV PY 2000 VL 28 IS 11 BP 3770 EP 3771 DI 10.1097/00003246-200011000-00053 PG 2 WC Critical Care Medicine SC General & Internal Medicine GA 376AQ UT WOS:000165436000043 PM 11098999 ER PT J AU Barrett, AJ AF Barrett, AJ TI Conditioning regimens for allogeneic stem cell transplants SO CURRENT OPINION IN HEMATOLOGY LA English DT Article ID BONE-MARROW TRANSPLANTATION; MIXED HEMATOPOIETIC CHIMERISM; CHRONIC MYELOID-LEUKEMIA; TOTAL-BODY IRRADIATION; PHARMACOLOGICAL IMMUNOSUPPRESSION; MOLECULAR REMISSION; THERAPY; DOGS; ALLOGRAFTS; ANTIGEN AB In addition to providing cytoreduction at myeloablative dose intensity, conditioning regimens for allogeneic transplantation are designed to immunosuppress the recipient to permit donor lymphohematopoietic engraftment and thereby establish a graft-versus-malignancy effect. Increased confidence in the potency of this allogeneic graft-versus-malignancy effect, together with the need to reduce dose intensity to make transplantation safer and more widely applicable in older patients, has led to a conceptual revolution in conditioning regimen design. Novel nonmyeloablative transplant conditioning treatments have low regimen-related toxicity and low transplant-related mortality. The transplants confer a graft-versus-malignancy effect in myeloid and lymphoid malignancies and in metastatic renal cell cancer. Future prospects are for low toxicity conditioning regimens combined with specific antileukemia or antitumor intensification with radioconjugated or unmodified antibodies and the application of highly immunosuppressive but low toxicity conditioning regimens for mismatched transplants. Curr Opin Hematol 2000, 7:339-342 (C) 2000 Lippincott Williams & Wilkins, Inc. C1 NHLBI, Hematol Branch, Allogene Stem Cell Transplant Sect, NIH, Bethesda, MD 20892 USA. RP Barrett, AJ (reprint author), NHLBI, Hematol Branch, Allogene Stem Cell Transplant Sect, NIH, Bldg 10,Room 7C 103,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 27 TC 15 Z9 15 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1065-6251 J9 CURR OPIN HEMATOL JI Curr. Opin. Hematol. PD NOV PY 2000 VL 7 IS 6 BP 339 EP 342 DI 10.1097/00062752-200011000-00003 PG 4 WC Hematology SC Hematology GA 465FV UT WOS:000170579700003 PM 11055505 ER PT J AU Shamim, EA Rider, LG Miller, FW AF Shamim, EA Rider, LG Miller, FW TI Update on the genetics of the idiopathic inflammatory myopathies SO CURRENT OPINION IN RHEUMATOLOGY LA English DT Review ID INCLUSION-BODY MYOSITIS; MAJOR HISTOCOMPATIBILITY COMPLEX; PENICILLAMINE-INDUCED MYOSITIS; JUVENILE RHEUMATOID-ARTHRITIS; SYSTEMIC LUPUS-ERYTHEMATOSUS; NECROSIS-FACTOR-ALPHA; CLASS-II ALLELES; ANTIGEN PM-SCL; AUTOIMMUNE-DISEASES; IMMUNOGENETIC FEATURES AB A number of lines of investigation suggest that, as is likely the case for other autoimmune diseases, the idiopathic inflammatory myopathies (IIM) develop as a result of specific environmental exposures in genetically susceptible individuals. Current data imply that multiple genes are involved in the etiology of these complex disorders. Targeted gene studies and whole genome approaches have begun to identify several genetic risk factors for autoimmune diseases, but the rarity and heterogeneity of the IIM have limited our knowledge of their associated genes. Current findings suggest that human leukocyte antigen (HLA) genes on chromosome 6, particularly HLA DRB1*0301 and the linked allele DQA1*0501, have the strongest associations with all clinical forms of IIM in white patients. Different HLA alleles, however, may confer risk or protection for myositis in distinct ethnic, serologic, and environmental exposure groups. Non-HLA genetic risk factors, which have been documented for other autoimmune diseases, are now being identified for the IIM. These include polymorphic genes encoding immunoglobulin heavy chains (defined by serologic markers known as Gm allotypes), cytokines and their receptors, and certain proteins that accumulate in the myocyte vacuoles of inclusion body myositis patients. Selected allelic polymorphisms of interleukin-1 receptor antagonist variable number tandem repeats and genes for tumor necrosis factor alpha and interleukin-l alpha also have recently been associated with IIM. The pathogenic bases for the differences among the many clinically, pathologically and immunologically defined syndromes known as the IIM will be elucidated through a better understanding of the multiple genes that define risks for their development, as well as through investigations of gene-gene and gene-environment interactions. (C) 2000 Lippincott Williams & Wilkins. Inc. C1 US FDA, Ctr Biol Evaluat & Res, Div Monoclonal Antibodies, Lab Mol & Dev Immunol, Bethesda, MD 20892 USA. RP Shamim, EA (reprint author), US FDA, Ctr Biol Evaluat & Res, Div Monoclonal Antibodies, Lab Mol & Dev Immunol, NIH Bldg 29B,Room 2G11,HFM 561,8800 Rockville Pik, Bethesda, MD 20892 USA. OI Rider, Lisa/0000-0002-6912-2458; Miller, Frederick/0000-0003-2831-9593 NR 87 TC 59 Z9 60 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1040-8711 J9 CURR OPIN RHEUMATOL JI CURR. OPIN. RHEUMATOL. PD NOV PY 2000 VL 12 IS 6 BP 482 EP 491 DI 10.1097/00002281-200011000-00002 PG 10 WC Rheumatology SC Rheumatology GA 367GW UT WOS:000090054200002 PM 11092196 ER PT J AU Lafont, BAP Gloeckler, L D'Hautcourt, JL Gut, JP Aubertin, AM AF Lafont, BAP Gloeckler, L D'Hautcourt, JL Gut, JP Aubertin, AM TI One-round determination of seven leukocyte subsets in rhesus macaque blood by flow cytometry SO CYTOMETRY LA English DT Article DE Macaca mulatta; flow cytometry; blood formula; lymphocyte subsets; SHIV ID SIMIAN IMMUNODEFICIENCY VIRUS; SIMULTANEOUS IDENTIFICATION; ENVELOPE GLYCOPROTEINS; HIV TYPE-1; MONKEYS; INFECTION; AIDS; SIVMAC; ENV; IMMUNOFLUORESCENCE AB Background: Rhesus macaques are frequently used in biomedical research as experimental models for studying infectious diseases and for preclinical vaccination trials. The infection of these monkeys with simian immunodeficiency viruses (SIV) or simian-human immunodeficiency viruses (SHIV) reproduces the clinical and immunological characteristics of human infection by human immunodeficiency virus (HIV) Evolution of the immune response in the infected animals is generally analyzed by determining the lymphocyte subsets on blood samples using now cytometry but requiring multiple, blood consuming, determinations. Methods: Cell subsets present in whole-blood samples were labeled with a combination of anti-human monoclonal antibodies to CD2, CD20, CD4, CD8, and CD14 coupled to FITC or PE and analyzed by flow cytometry. Results: In one round, we obtained the precise determination of macaque blood cell composition by flow cytometry. Monocytes, granulocytes, eosinophils, B lymphocytes, helper, and cytotoxic T lymphocytes were distinguished. Results obtained correlated strongly with those obtained with conventional blood cell differential systems and with separate staining of lymphocytes. The analysis of blood from healthy rhesus macaques and SHIV-infected animals demonstrated the accuracy of the determination even in very pathological situations such as macaques with simian IUDS. Conclusions: Our method allows fast determination of the blood cell composition and will be particularly useful to evaluate the cell subset evolution of macaques involved in large-scale experimental trials. (C) 2000 Wiley-Liss, Inc. C1 INSERM, Unite Rech 74, Inst Virol, Strasbourg, France. Hop Warquignies, CHR Clin St Joseph, Lab Cytometrie, Boussu, Belgium. RP Lafont, BAP (reprint author), NIAID, Mol Microbiol Lab, NIH, Bldg 4,Room 328,4 Ctr Dr, Bethesda, MD 20892 USA. RI Lafont, Bernard/B-7236-2014 NR 24 TC 11 Z9 11 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 0196-4763 J9 CYTOMETRY JI Cytometry PD NOV 1 PY 2000 VL 41 IS 3 BP 193 EP 202 DI 10.1002/1097-0320(20001101)41:3<193::AID-CYTO6>3.0.CO;2-G PG 10 WC Biochemical Research Methods; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 368LX UT WOS:000090119400006 PM 11042616 ER PT J AU Soyal, SM Amleh, A Dean, J AF Soyal, SM Amleh, A Dean, J TI FIG alpha, a germ cell-specific transcription factor required for ovarian follicle formation SO DEVELOPMENT LA English DT Article DE primordial follicles; germ-line transcription factor; factor in the germline; alpha (FIG alpha) ID ZONA-PELLUCIDA GENES; CHROMOSOME SYNAPSIS; COORDINATE EXPRESSION; MOUSE EMBRYO; STEM-CELLS; MICE; DISRUPTION; MEIOSIS; FAILURE; DIFFERENTIATION AB Primordial follicles are formed perinatally in mammalian ovaries and at birth represent the lifetime complement of germ cells, With cyclic periodicity, cohorts enter into a growth phase that culminates in ovulation of mature eggs, but little is known about the regulatory cascades that govern these events. FIG alpha, a transcription factor implicated in postnatal oocyte-specific gene expression, is detected as early as embryonic day 13, Mouse lines lacking FIG alpha were established by targeted mutagenesis in embryonic stem cells. Although embryonic gonadogenesis appeared normal, primordial follicles were not formed at birth, and massive depletion of oocytes resulted in shrunken ovaries and female sterility. Fig alpha (the gene for FIG alpha) null males have normal fertility. The additional observation that null females do not express Zp1, Zp2 or Zp3 indicates that FIG alpha plays a key regulatory role in the expression of multiple oocyte-specific genes, including those that initiate folliculogenesis and those that encode the zona pellucida required for fertilization and early embryonic survival. The persistence of FIG alpha in adult females suggests that it may regulate additional pathways that are essential for normal ovarian development. C1 NIDDK, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Soyal, SM (reprint author), NIDDK, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NR 46 TC 300 Z9 314 U1 0 U2 5 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD NOV PY 2000 VL 127 IS 21 BP 4645 EP 4654 PG 10 WC Developmental Biology SC Developmental Biology GA 376PF UT WOS:000165465400012 PM 11023867 ER PT J AU Wagner, KU Claudio, E Rucker, EB Riedlinger, G Broussard, C Schwartzberg, PL Siebenlist, U Hennighausen, L AF Wagner, KU Claudio, E Rucker, EB Riedlinger, G Broussard, C Schwartzberg, PL Siebenlist, U Hennighausen, L TI Conditional deletion of the Bcl-x gene from erythroid cells results in hemolytic anemia and profound splenomegaly SO DEVELOPMENT LA English DT Article DE Bcl-x; Bax; Bcl2 family; erythropoiesis; hyperplasia; MMTV-LTR; Cre recombinase ID TRANSCRIPTION FACTOR GATA-1; EMBRYONIC STEM-CELLS; TRANSGENIC MICE; DEFICIENT MICE; DEATH; BCL-X(L); ERYTHROPOIETIN; EXPRESSION; BAX; DIFFERENTIATION AB Bcl-x is a member of the Bcl2 family and has been suggested to be important for the survival and maturation of various cell types including the erythroid lineage. To define the consequences of Bcl-x loss in erythroid cells and other adult tissues, we have generated mice conditionally deficient in the Bcl-x gene using the Cre-loxP recombination system. The temporal and spatial excision of the flexed Bcl-x locus was achieved by expressing the Cre recombinase gene under control of the MMTV-LTR. By the age of five weeks, Bcl-x conditional mutant mice exhibited hyperproliferation of megakaryocytes and a decline in the number of circulating platelets. Three-month-old animals suffered from severe hemolytic anemia, hyperplasia of immature erythroid cells and profound enlargement of the spleen. We demonstrate that Bcl-x is only required for the survival of erythroid cells at the end of maturation, which includes enucleated reticulocytes in circulation. The extensive proliferation of immature erythroid cells in the spleen and bone marrow might be the result of a fast turnover of late red blood cell precursors and accelerated erythropoiesis in response to tissue hypoxia. The increase in cell death of late erythroid cells is independent from the proapoptotic factor Bax, as demonstrated in conditional double mutant mice for Bcl-x and Bax. Mice conditionally deficient in Bcl-x permitted us for the first time to study the effects of Bcl-x deficiency on cell proliferation, maturation and survival under physiological conditions in an adult animal. C1 NIDDKD, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, Immune Activat Sect, NIH, Bethesda, MD 20892 USA. NHGRI, Cell Signaling Sect, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. RP Wagner, KU (reprint author), Univ Nebraska, Med Ctr, Eppley Inst Res Canc & Allied Dis, Nebraska Med Ctr 986805, Rm 8009, Omaha, NE 68198 USA. RI Wagner, Kay-Uwe/B-6044-2009; Brugnara, Carlo/A-8041-2010 OI Brugnara, Carlo/0000-0001-8192-8713 NR 37 TC 129 Z9 132 U1 1 U2 3 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD NOV PY 2000 VL 127 IS 22 BP 4949 EP 4958 PG 10 WC Developmental Biology SC Developmental Biology GA 381GN UT WOS:000165754100019 PM 11044408 ER PT J AU Liu, C Knezevic, V Hunter, S Thompson, K Mackem, S AF Liu, C Knezevic, V Hunter, S Thompson, K Mackem, S TI Regulation of limb pattern: New roles for old factors? SO DEVELOPMENTAL DYNAMICS LA English DT Meeting Abstract C1 NCI, Pathol Lab, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1058-8388 J9 DEV DYNAM JI Dev. Dyn. PD NOV PY 2000 VL 219 IS 3 MA 2 BP 429 EP 429 PG 1 WC Anatomy & Morphology; Developmental Biology SC Anatomy & Morphology; Developmental Biology GA 370KL UT WOS:000165122200016 ER PT J AU Clocquet, AR Egan, JM Stoffers, DA Muller, DC Wideman, L Chin, GA Clarke, WL Hanks, JB Habener, JF Elahi, D AF Clocquet, AR Egan, JM Stoffers, DA Muller, DC Wideman, L Chin, GA Clarke, WL Hanks, JB Habener, JF Elahi, D TI Impaired insulin secretion and increased insulin sensitivity in familial maturity-onset diabetes of the young 4 (Insulin promoter factor 1 gene) SO DIABETES LA English DT Article ID NUCLEAR FACTOR-1-ALPHA GENE; PANCREATIC BETA-CELLS; GLUCOKINASE GENE; GLUCOSE; MUTATIONS; TRANSCRIPTION; MELLITUS; DEFECT; MODY3; ACTIVATION AB Diabetes resulting from heterozygosity for an inactivating mutation of the homeodomain transcription factor insulin promoter factor 1 (IPF-1) is due to a genetic defect of beta -cell function referred to as maturity-onset diabetes of the young 4, IPF-1 is required for the development of the pancreas and mediates glucose-responsive stimulation of insulin gene transcription. To quantitate islet cell responses in a family harboring a Pro63fsdelC mutation in IPF-1, we performed a five-step (1-h intervals) hyperglycemic clamp on seven heterozygous members (NM) and eight normal genotype members (NN), During the last 30 min of the fifth glucose step, glucagon-like peptide 1 (GLP-1) was also infused (1.5 pmol . kg(-1) . min(-1)). Fasting plasma glucose levels were greater in the NM group than in the NN group (9.2 vs. 5.9 mmol/l, respectively; P < 0.05). Fasting insulin levels were similar in both groups (72 vs. 105 pmol/l for NN vs. NM, respectively). First-phase insulin and C-peptide responses were absent in individuals in the NM group, who had markedly attenuated insulin responses to glucose alone compared with the NN group, At a glucose level of 16.8 mmol/l above fasting level, GLP-1 augmented insulin secretion equivalently (fold increase) in both groups, but the insulin and C-peptide responses to GLP-1 were sevenfold less in the NM subjects than in the NN subjects. In both groups, glucagon levels fell during each glycemic plateau, and a further reduction occurred during the GLP-1 infusion, Sigmoidal dose-response curves of glucose clearance versus insulin levels during the hyperglycemic clamp in the two small groups showed both a left shift and a lower maximal response in the NM group compared with the NN group, which is consistent with an increased insulin sensitivity in the NM subjects, A sharp decline occurred in the dose-response curve for suppression of nonesterified fatty acids versus insulin levels in the NM group. We conclude that the Pro63fsdelC IPF-1 mutation is associated with a severe impairment of -cell sensitivity to glucose and an apparent increase in peripheral tissue sensitivity to insulin and is a genetically determined cause of beta -cell dysfunction. C1 Massachusetts Gen Hosp, Geriatr Res Lab, Boston, MA 02114 USA. Massachusetts Gen Hosp, Mol Endocrinol Lab, Boston, MA 02114 USA. Harvard Univ, Sch Med, Howard Hughes Med Inst, Boston, MA 02115 USA. NIA, Clin Invest Lab, NIH, Baltimore, MD 21224 USA. Univ Virginia, Hlth Sci Ctr, Dept Med, Charlottesville, VA USA. Univ Virginia, Hlth Sci Ctr, Dept Pediat, Charlottesville, VA USA. Univ Virginia, Hlth Sci Ctr, Dept Surg, Charlottesville, VA 22908 USA. RP Elahi, D (reprint author), Massachusetts Gen Hosp, Geriatr Res Lab, GRB SB 0015,55 Fruit St, Boston, MA 02114 USA. RI Stoffers, Doris/H-6157-2012 FU NIA NIH HHS [AG-00599]; NIDDK NIH HHS [DK-30457, DK-30834] NR 37 TC 44 Z9 49 U1 0 U2 1 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD NOV PY 2000 VL 49 IS 11 BP 1856 EP 1864 DI 10.2337/diabetes.49.11.1856 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 370LC UT WOS:000165123700013 PM 11078452 ER PT J AU Kubota, N Tobe, K Terauchi, Y Eto, K Yamauchi, T Suzuki, R Tsubamoto, Y Komeda, K Nakano, I Miki, H Satoh, S Sekihara, H Sciacchitano, S Lesniak, M Aizawa, S Nagai, R Kimura, S Akanuma, Y Taylor, SI Kadowaki, T AF Kubota, N Tobe, K Terauchi, Y Eto, K Yamauchi, T Suzuki, R Tsubamoto, Y Komeda, K Nakano, I Miki, H Satoh, S Sekihara, H Sciacchitano, S Lesniak, M Aizawa, S Nagai, R Kimura, S Akanuma, Y Taylor, SI Kadowaki, T TI Disruption of insulin receptor substrate 2 causes type 2 diabetes because of liver insulin resistance and lack of compensatory beta-cell hyperplasia SO DIABETES LA English DT Article ID STIMULATES TYROSINE PHOSPHORYLATION; NADH SHUTTLE SYSTEM; GROWTH FACTOR-I; PHOSPHOTYROSINE PROTEIN; MICE LACKING; IRS-2; SECRETION; ACTIVATION; KNOCKOUT; PATHWAY AB To investigate the role of insulin receptor substrate (IRS)-2 in vivo, we generated IRS-2-deficient mice by gene targeting. Although homozygous IRS-2-deficient mice (IRS-2(-/-) mice) had a body weight similar to wildtype mice, they progressively developed type 2 diabetes at 10 weeks. IRS-2(-/-) mice showed insulin resistance and a defect in the insulin-stimulated signaling pathway in liver but not in skeletal muscle. Despite insulin resistance, the amount of beta -cells was reduced to 83% of that in wild-type mice, which was in marked contrast to the 85% increase in the amount of beta -cells in IRS-1-deficient mice (IRS-1(-/-) mice) to compensate for insulin resistance. Thus, IRS-2 plays a crucial role in the regulation of beta -cell mass. On the other hand, insulin secretion by the same number of cells in response to glucose measured ex vivo was significantly increased in IRS-2(-/-) mice compared with wild-type mice but was decreased in IRS-1(-/-) mice. These results suggest that IRS-1 and IRS-S may play different roles in the regulation of beta -cell mass and the function of individual beta -cells. C1 Univ Tokyo, Grad Sch Med, Dept Metab Dis, Bunkyo Ku, Tokyo 1138655, Japan. Tokyo Med Univ, Anim Res Ctr, Div Lab Anim Sci, Tokyo, Japan. Yokohama City Univ, Dept Internal Med 3, Yokohama, Kanagawa 232, Japan. Asahi Life Fdn, Inst Diabet Care & Res, Kumamoto, Japan. Kumamoto Univ, Sch Med, Inst Mol Embryol & Genet, Dept Morphogenesis, Kumamoto 860, Japan. Natl Inst Diabet & Digest Dis, Diabet Branch, NIH, Bethesda, MD USA. RP Kadowaki, T (reprint author), Univ Tokyo, Grad Sch Med, Dept Metab Dis, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1138655, Japan. RI Kubota, Naoto/N-7892-2015 NR 30 TC 327 Z9 350 U1 1 U2 11 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD NOV PY 2000 VL 49 IS 11 BP 1880 EP 1889 DI 10.2337/diabetes.49.11.1880 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 370LC UT WOS:000165123700016 PM 11078455 ER PT J AU Gavrilova, O Marcus-Samuels, B Reitman, ML AF Gavrilova, O Marcus-Samuels, B Reitman, ML TI Lack of Responses to a beta(3)-adrenergic agonist in lipoatrophic A-ZIP/F-1 mice SO DIABETES LA English DT Article ID BROWN ADIPOSE-TISSUE; YELLOW KK MICE; BETA-ADRENOCEPTOR; TRANSGENIC MICE; BETA-3-ADRENERGIC RECEPTORS; INSULIN-SECRETION; MESSENGER-RNA; CL-316,243; OBESITY; ANTIOBESITY AB Stimulation of beta (3)-adrenergic receptors increases metabolic rate via lipolysis in white adipose tissue (WAT) and thermogenesis in brown adipose tissue (BAT), Other acute effects include decreased gastrointestinal motility and food intake and increased insulin secretion. Chronic treatment with a beta (3) agonist ameliorates diabetes and obesity in rodents. We studied the effects of beta (3) stimulation in A-ZIP/F-1 mice, which have virtually no WAT, a reduced amount of BAT, severe insulin resistance, and diabetes. In contrast with wild-type mice, treatment of A-ZIP/F-1 mice with CL316243, a beta (3)-adrenergic agonist, did not increase O-2 consumption. A single dose of CL316243 produced a 2-fold increase in serum free fatty acids, a 53-fold increase in insulin, and a 2.4-fold decrease in glucose levels in wild-type mice but no change in A-ZIP/F-1 animals. The A-ZIP/F-1 mice also did not show reduced gastrointestinal motility or 24-h food intake during beta (3) stimulation. Chronic administration of CL316243 to the A-ZIP/F-1 mice did not improve their thermogenesis, hyperglycemia, or hyperinsulinemia, Thus, all of the beta (3) effects studied were absent in the lipoatrophic A-ZIP/F-1 mice, including the effects on nonadipose tissues. From these results, we suggest that all of the effects of beta (3) agonists are initiated at the adipocyte with the nonadipose effects being secondary events presumably mediated by signals from adipose tissue. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Gavrilova, O (reprint author), NIDDKD, Diabet Branch, NIH, Bldg 10,Room 8N-250, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013 OI Reitman, Marc/0000-0002-0426-9475 NR 38 TC 40 Z9 40 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD NOV PY 2000 VL 49 IS 11 BP 1910 EP 1916 DI 10.2337/diabetes.49.11.1910 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 370LC UT WOS:000165123700020 PM 11078459 ER PT J AU Bray, GA Culbert, IW Champagne, CM Dawson, L Eberhardt, B Greenway, FL Guillory, FG Hebert, AA Jeffirs, ML Kennedy, BM Lovejoy, JC Melancon, LE Morris, LH Reed, L Perault, J Ryan, DH Sanford, DA Smith, KG Smith, LL St Amant, JA Tucker, E Tulley, RT Vicknair, PC Williamson, D Zachwieja, JJ Tobian, JA Matulik, MJ Clarke, B Collins, DA Czech, KB DeSandre, C Ehrmann, DA Geiger, G Harding-Clay, B Hilbrich, RM McNabb, WL Polonsky, KS Semenske, AR Stepp, KA Watson, PG Mendoza, JT Smith, KA Goldstein, BJ Lark, C Liberoni, R Murphy, L Pepe, C Spandorfer, JM Goldberg, RB Rowe, P Calles, J Casanova, P Donahue, RP Florez, HJ Giannella, A McLymont, V Mendez, J O'Hara, P Ojito, J Prineas, R Haffner, SM Montez, MG Miettinen, H Mobley, CM Mykkanen, LA Rozek, MM Hamman, RF Nash, PV Calonge, BN Hill, JO Hoyer, SR Jortberg, BT Miller-Stone, M Regensteiner, JG Seagle, H Steinke, SC Testaverde, L Van-Dorsten, B Horton, ES Lawton, KE Arky, RA Bryant, M Burke, JP Caballero, E Callaghan, KM Ganda, OP Franklin, T Jackson, SD Jacobsen, AM Kula, LM Kocal, M Malloy, MA Nicosia, M Oldmixon, CF Pan, J Quitingon, M Rubtchinsky, S Seely, EW Schweizer, D Simonson, D Smith, F Solomon, CG Warram, J Kahn, SE Montgomery, BK Fujimoto, WY Knopp, RH Lipkin, EW Marr, M McCann, BS Nguyen, TT Palmer, JP Schwartz, RS Kitabchi, AE Murphy, ME Applegate, WB Bryer-Ash, M Crisler, A Cunningham, G Franklin, AW Frieson, SL Green, DL Imseis, R Kennedy, CL Lambeth, HC Lichtermann, LC Oktaei, H O'Toole, ML Ricks, H Rutledge, LMK Sherman, AR Smith, CM Soberman, JE Williams-Cleaves, BJ Metzger, BE Johnson, MK Behrends, C Cook, ML Fitzgibbon, M Heard, D McPherson, D Molitch, ME Moore, M Pitts, T Roston, S Schinleber, PA Nathan, DM McKitrick, C Abbott, K Anderson, E Bissett, L Cagliero, E Crowell, S Delahanty, L Fritz, S Levina, E Michel, T Norman, D O'Keefe, J Poulos, A Ronan, L Rosal, M Salerno, M Shagensky, C Steiner, B Turgeon, H Young, A Olefsky, JM Carrion-Petersen, ML Barrett-Connor, E Beltran, M Caenepeel-Mills, K Edelman, SV Ford, RO Garcia, J Henry, RR Hill, M Horne, J Leos, D Mudaliar, S Pollard, A Torio-Hurley, J Pi-Sunyer, FX Lee, JE Allison, DB Aronoff, NJ Barreras, IM Crandall, JP Orlando, SJ Parkes, K Rooney, ES Van Wye, GEH Viscovich, KA Marrero, DG Kukman-Kelly, MS Dotson, YF Fineberg, SE Guare, JC Hadden, A Ignaut, JM Jackson, MA Kirkman, MS Porter, BD Prince, MJ Wheeler, ML Ratner, RE Youssef, G Boner, A Bronsord, M Brown, E Cheatham, WW Cola, S Comfort, A Boggs, G Evans, C Levetan, C Kellum, T Nair, AK Passaro, MD Rubin, R Shapiro, S Saad, MF Budgett, M Jinagouda, SD Aziz, A Bernaba, B Bodkin, SL Ciobanu, V Cosenza, C Hagar, JE Khan, A Kumar, D Khawaja, Q Lui, G Marston, AR Mehta, V Sharma, AR Szamos, K Vargas, A Vargas, B Zambrana, N Dagogo-Jack, S Santiago, AS Brendle, JS Fisher, EB Gherardini, DC Heins, JM Marsala, JM Raspberry, CC Santiago, JV Saudek, CD Bradley, VL Brancati, FL Cappelli, S Charleston, JB Rubin, RR Stewart, KJ Sullivan, E Schade, DS Adams, KS Atler, LF Bowling, DA Boyle, PJ Burge, MR Butler, L Canady, JL Chai, L Guillen, M Gutierrez, M Hornbeck, D Johannes, C Katz, P King, C McCalman, R Montoya, DA Rassam, A Senter, W Shamoon, H Brown, JO Adames, J Blanco, E Cox, L Duffy, H Engel, S Friedler, A Harroun, T Howard-Century, CJ Kloiber, S Longchamp, N Pompi, D Walker, EA Wylie-Rosett, J Zonszein, J Wing, RR Kramer, MK Barr, S Clifford, L Culyba, R Frazier, M Gilligan, R Harris, L Harrier, S Henderson, W Jeffries, S Koenning, G Maholic, K Mullen, M Noel, A Orchard, TJ Otto, A Smith, M Stapinski, V Viteri, J Wilson, T Williams, KV Zgibor, J Arakaki, RF Latimer, RW Baker-Ladao, NK Beddow, RM Dias, LM Dupont, DA Fukuhara, LL Mau, MK Odom, SK Perry, RU Tokunaga, JS Knowler, WC Cooeyate, NJ Hoskin, MA Percy, CA Acton, KJ Andre, VL Antone, S Baptisto, NM Barber, R Bennett, PH Benson, MB Bird, EC Broussard, BA Chavez, M Dacawyma, TS Duncan, R Ghahate, JM Glass, M Gohdes, D Grant, G Hanson, RL Horse, E Hughte, G Ingraham, LE Jackson, MC Jay, PA Kaskalla, RS Kessler, D Kobus, KM Morgan, T Nashboo, Y Poirier, S Reidy, M Ridpath, S Roumain, J Rowse, DH Roy, RJ Sangster, S Sewemaenewa, J Wilson, C Yazzie, M Fowler, S Bain, R Bamdad, J Brenneman, T Callaghan, J Edelstein, SL Grimes, KL Jones, S Jones, TL Lachin, JM Mucik, P Orlosky, R Stimpson, CE Suiter, C Temprosa, E Van Aerden, C Eastman, R Garfield, S Andres, R Engelgau, MM Narayan, KMV Williamson, DF Herman, WH Marcovina, SM Aldrich, A Chandler, WL Rautaharju, PM Alexander, T Pemberton, NT Prineas, R Rautaharju, FSR Zhang, Z Mayer-Davis, EJ Martin, M O'Leary, DH Funk, LRC O'Leary, KA Polak, JF Stamm, ER Scherzinger, AL Wing, RR Gillis, BP Kriska, AM Huffmyer, C Venditti, EM Kaplan, RM David, KM Sieber, WJ AF Bray, GA Culbert, IW Champagne, CM Dawson, L Eberhardt, B Greenway, FL Guillory, FG Hebert, AA Jeffirs, ML Kennedy, BM Lovejoy, JC Melancon, LE Morris, LH Reed, L Perault, J Ryan, DH Sanford, DA Smith, KG Smith, LL St Amant, JA Tucker, E Tulley, RT Vicknair, PC Williamson, D Zachwieja, JJ Tobian, JA Matulik, MJ Clarke, B Collins, DA Czech, KB DeSandre, C Ehrmann, DA Geiger, G Harding-Clay, B Hilbrich, RM McNabb, WL Polonsky, KS Semenske, AR Stepp, KA Watson, PG Mendoza, JT Smith, KA Goldstein, BJ Lark, C Liberoni, R Murphy, L Pepe, C Spandorfer, JM Goldberg, RB Rowe, P Calles, J Casanova, P Donahue, RP Florez, HJ Giannella, A McLymont, V Mendez, J O'Hara, P Ojito, J Prineas, R Haffner, SM Montez, MG Miettinen, H Mobley, CM Mykkanen, LA Rozek, MM Hamman, RF Nash, PV Calonge, BN Hill, JO Hoyer, SR Jortberg, BT Miller-Stone, M Regensteiner, JG Seagle, H Steinke, SC Testaverde, L Van-Dorsten, B Horton, ES Lawton, KE Arky, RA Bryant, M Burke, JP Caballero, E Callaghan, KM Ganda, OP Franklin, T Jackson, SD Jacobsen, AM Kula, LM Kocal, M Malloy, MA Nicosia, M Oldmixon, CF Pan, J Quitingon, M Rubtchinsky, S Seely, EW Schweizer, D Simonson, D Smith, F Solomon, CG Warram, J Kahn, SE Montgomery, BK Fujimoto, WY Knopp, RH Lipkin, EW Marr, M McCann, BS Nguyen, TT Palmer, JP Schwartz, RS Kitabchi, AE Murphy, ME Applegate, WB Bryer-Ash, M Crisler, A Cunningham, G Franklin, AW Frieson, SL Green, DL Imseis, R Kennedy, CL Lambeth, HC Lichtermann, LC Oktaei, H O'Toole, ML Ricks, H Rutledge, LMK Sherman, AR Smith, CM Soberman, JE Williams-Cleaves, BJ Metzger, BE Johnson, MK Behrends, C Cook, ML Fitzgibbon, M Heard, D McPherson, D Molitch, ME Moore, M Pitts, T Roston, S Schinleber, PA Nathan, DM McKitrick, C Abbott, K Anderson, E Bissett, L Cagliero, E Crowell, S Delahanty, L Fritz, S Levina, E Michel, T Norman, D O'Keefe, J Poulos, A Ronan, L Rosal, M Salerno, M Shagensky, C Steiner, B Turgeon, H Young, A Olefsky, JM Carrion-Petersen, ML Barrett-Connor, E Beltran, M Caenepeel-Mills, K Edelman, SV Ford, RO Garcia, J Henry, RR Hill, M Horne, J Leos, D Mudaliar, S Pollard, A Torio-Hurley, J Pi-Sunyer, FX Lee, JE Allison, DB Aronoff, NJ Barreras, IM Crandall, JP Orlando, SJ Parkes, K Rooney, ES Van Wye, GEH Viscovich, KA Marrero, DG Kukman-Kelly, MS Dotson, YF Fineberg, SE Guare, JC Hadden, A Ignaut, JM Jackson, MA Kirkman, MS Porter, BD Prince, MJ Wheeler, ML Ratner, RE Youssef, G Boner, A Bronsord, M Brown, E Cheatham, WW Cola, S Comfort, A Boggs, G Evans, C Levetan, C Kellum, T Nair, AK Passaro, MD Rubin, R Shapiro, S Saad, MF Budgett, M Jinagouda, SD Aziz, A Bernaba, B Bodkin, SL Ciobanu, V Cosenza, C Hagar, JE Khan, A Kumar, D Khawaja, Q Lui, G Marston, AR Mehta, V Sharma, AR Szamos, K Vargas, A Vargas, B Zambrana, N Dagogo-Jack, S Santiago, AS Brendle, JS Fisher, EB Gherardini, DC Heins, JM Marsala, JM Raspberry, CC Santiago, JV Saudek, CD Bradley, VL Brancati, FL Cappelli, S Charleston, JB Rubin, RR Stewart, KJ Sullivan, E Schade, DS Adams, KS Atler, LF Bowling, DA Boyle, PJ Burge, MR Butler, L Canady, JL Chai, L Guillen, M Gutierrez, M Hornbeck, D Johannes, C Katz, P King, C McCalman, R Montoya, DA Rassam, A Senter, W Shamoon, H Brown, JO Adames, J Blanco, E Cox, L Duffy, H Engel, S Friedler, A Harroun, T Howard-Century, CJ Kloiber, S Longchamp, N Pompi, D Walker, EA Wylie-Rosett, J Zonszein, J Wing, RR Kramer, MK Barr, S Clifford, L Culyba, R Frazier, M Gilligan, R Harris, L Harrier, S Henderson, W Jeffries, S Koenning, G Maholic, K Mullen, M Noel, A Orchard, TJ Otto, A Smith, M Stapinski, V Viteri, J Wilson, T Williams, KV Zgibor, J Arakaki, RF Latimer, RW Baker-Ladao, NK Beddow, RM Dias, LM Dupont, DA Fukuhara, LL Mau, MK Odom, SK Perry, RU Tokunaga, JS Knowler, WC Cooeyate, NJ Hoskin, MA Percy, CA Acton, KJ Andre, VL Antone, S Baptisto, NM Barber, R Bennett, PH Benson, MB Bird, EC Broussard, BA Chavez, M Dacawyma, TS Duncan, R Ghahate, JM Glass, M Gohdes, D Grant, G Hanson, RL Horse, E Hughte, G Ingraham, LE Jackson, MC Jay, PA Kaskalla, RS Kessler, D Kobus, KM Morgan, T Nashboo, Y Poirier, S Reidy, M Ridpath, S Roumain, J Rowse, DH Roy, RJ Sangster, S Sewemaenewa, J Wilson, C Yazzie, M Fowler, S Bain, R Bamdad, J Brenneman, T Callaghan, J Edelstein, SL Grimes, KL Jones, S Jones, TL Lachin, JM Mucik, P Orlosky, R Stimpson, CE Suiter, C Temprosa, E Van Aerden, C Eastman, R Garfield, S Andres, R Engelgau, MM Narayan, KMV Williamson, DF Herman, WH Marcovina, SM Aldrich, A Chandler, WL Rautaharju, PM Alexander, T Pemberton, NT Prineas, R Rautaharju, FSR Zhang, Z Mayer-Davis, EJ Martin, M O'Leary, DH Funk, LRC O'Leary, KA Polak, JF Stamm, ER Scherzinger, AL Wing, RR Gillis, BP Kriska, AM Huffmyer, C Venditti, EM Kaplan, RM David, KM Sieber, WJ CA Diabet Prevention Program Res Grp TI The Diabetes Prevention Program - Baseline characteristics of the randomized cohort SO DIABETES CARE LA English DT Article ID IMPAIRED GLUCOSE-TOLERANCE; 10-YEAR FOLLOW-UP; FAT DISTRIBUTION; PIMA-INDIANS; MELLITUS; INSULIN; MEN; NIDDM; DIET; CLASSIFICATION AB OBJECTIVE - The Diabetes Prevention Program (DPP) is a 27-center randomized clinical trial designed to evaluate the safety and efficacy of interventions that may delay or prevent development of diabetes in people at increased risk for type 2 diabetes. RESEARCH DESIGN AND METHODS - Eligibility requirements were age greater than or equal to 25 years, BMI greater than or equal to 24 kg/m(2) (greater than or equal to 22 kg/m(2) for Asian-Americans), and impaired glucose tolerance plus a fasting plasma glucose of 5.3-6.9 mmol/l (or less than or equal to6.9 mmol for American Indians). Randomization of participants into the DPP over 2.7 years ended in June 1999. Baseline data for the three treatment groups-intensive lifestyle modification, standard care plus metformin, and standard care plus placebo-are presented for the 3,234 participants who have been randomized. RESULTS - Of all participants, 55% were Caucasian, 20% were African-American, 16% were Hispanic, 5% were American Indian, and 4% were Asian-American. Their average age at entry was 51 +/- 10.7 years (mean +/- SD), and 67.7% were women. Moreover, 16% were <40 years of age, and 20% were 60 years of age. Of the women, 48% were postmenopausal. Men and women had similar frequencies of history of hypercholesterolemia (37 and 33%, respectively) or hypertension (29 and 26%, respectively). On the basis of fasting lipid determinations, 54% of men and 40% of women fit National Cholesterol Education Program criteria for abnormal lipid profiles. More men than women were current or former cigarette smokers or had a history of coronary heart disease. Furthermore, 66% of men and 71% of women had a first-degree relative with diabetes. Overall, BMI averaged 34.0 +/- 6.7 kg/m(2) at baseline with 57% of the men and 73% of women having a BMI greater than or equal to 30 kg/m(2). Average fasting plasma glucose (6.0 +/- 0.5 mmol/l) and HbA(1c) (5.9 +/- 0.5%) in men were comparable with values in women (5.9 +/- 0.4 mmol/l and 5.9 +/- 0.5%, respectively). CONCLUSIONS - The DPP has successfully randomized a large cohort of participants with a wide distribution of age, obesity, and ethnic and racial backgrounds who are at high risk for developing type 2 diabetes. The study will examine the effects of interventions on the development of diabetes. C1 George Washington Univ, Diabet Prevent Program Coordinating Ctr, Rockville, MD 20852 USA. George Washington Univ, Ctr Biostat, Rockville, MD 20852 USA. Pennington Biomed Res Ctr, Baton Rouge, LA USA. Univ Chicago, Chicago, IL 60637 USA. Thomas Jefferson Univ, Jefferson Med Coll, Philadelphia, PA 19107 USA. Univ Miami, Coral Gables, FL 33124 USA. Univ Texas, Hlth Sci Ctr, San Antonio, TX 78284 USA. Univ Colorado, Boulder, CO 80309 USA. Joslin Diabet Ctr, Boston, MA 02215 USA. Univ Washington, Seattle, WA 98195 USA. Univ Tennessee, Knoxville, TN 37996 USA. Northwestern Univ, Sch Med, Evanston, IL 60208 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. St Lukes Roosevelt Hosp, New York, NY 10025 USA. Indiana Univ, Bloomington, IN 47405 USA. Medstar Res Inst, New York, NY USA. Univ Calif Los Angeles, Sch Med, Los Angeles, CA 90024 USA. Washington Univ, St Louis, MO USA. Johns Hopkins Univ, Sch Med, Baltimore, MD 21218 USA. Univ New Mexico, Sch Med, Albuquerque, NM 87131 USA. Albert Einstein Coll Med, Bronx, NY 10467 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. Univ Hawaii, Honolulu, HI 96822 USA. George Washington Univ, Ctr Biostat, Data Coordinating Ctr, Washington, DC 20052 USA. NIDDKD, Program Off, Bethesda, MD 20892 USA. NIA, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Atlanta, GA USA. Univ Michigan, Ann Arbor, MI 48109 USA. Wake Forest Univ, Sch Med, Epicare Ctr, Winston Salem, NC 27109 USA. Univ S Carolina, Nutr Coding Ctr, Columbia, SC 29208 USA. New England Med Ctr, Cent Carotid Ultrasound Unit, Boston, MA 02111 USA. Univ Colorado, Hlth Sci Ctr, CT Scan Reading Unit, Boulder, CO 80309 USA. Univ Calif San Diego, Qual Well Being Reading Unit, San Diego, CA 92103 USA. RP Bray, GA (reprint author), George Washington Univ, Diabet Prevent Program Coordinating Ctr, 6110 Execut Blvd,Suite 750, Rockville, MD 20852 USA. RI Narayan, K.M. Venkat /J-9819-2012 OI Narayan, K.M. Venkat /0000-0001-8621-5405 NR 48 TC 125 Z9 127 U1 0 U2 11 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD NOV PY 2000 VL 23 IS 11 BP 1619 EP 1629 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 368FD UT WOS:000090106300004 ER PT J AU Resnick, HE Vinik, AI Schwartz, AV Leveille, SG Brancati, FL Balfour, J Guralnik, JM AF Resnick, HE Vinik, AI Schwartz, AV Leveille, SG Brancati, FL Balfour, J Guralnik, JM TI Independent effects of peripheral nerve dysfunction on lower-extremity physical function in old age - The women's health and aging study SO DIABETES CARE LA English DT Article; Proceedings Paper CT 60th Scientific Sessions of the American-Diabetes-Association CY JUN 09-13, 2000 CL SAN ANTONIO, TEXAS SP Amer Diabet Assoc ID DIABETIC NEUROPATHY; BODY SWAY; DISABILITY; RELIABILITY; CONDUCTION; STRENGTH AB OBJECTIVE - To determine the role of peripheral nerve dysfunction (PND) in the disablement pathway RESEARCH DESIGN AND METHODS - Vibration perception threshold (VPT) was measured in 894 women aged greater than or equal to 65 years, and those with normal peripheral nerve function and with mild, moderate, and severe PND were identified Lower-extremity impairments included quadriceps strength (kilograms) and three progressively difficult balance tasks (able/unable). Functional limitations included rising from a chair (able/unable) and usual place and fast-paced walking speeds (meters/second). Level of PND was related to impairments and functional limitations in linear and logistic regression models that controlled for potentially confounding factors, including reported diabetes. RESULTS - Level of PND was associated with impaired balance (adjusted odds ratios: 2.21, 1.95, and 3.02 for mild, moderate, and severe PND, respectively, relative to normal, P < 0.05). PND was also associated with decrements in both usual and fast-paced walking speeds (-0.08, -0.08, and -0.15 m/s for usual pace and -0.13, -0.12, and -0.24m/s for fast-paced walking speed for women with mild, moderate, and severe PND, respectively, P < 0.01 for all). Reported diabetes was not associated with these outcomes in the presence of PND. Some, but nor all, of the association between PND and functional limitations was explained by the relationship between PND and impairments. CONCLUSIONS - PND is significantly associated with both lower-extremity impairments and functional limitations in older women, and PND appears to have independent effects on functional limitations. The independent effect of diabetes on these outcomes may be limited when PND is considered. Further research is needed to determine if PND is causally related to disability in old age. C1 NIA, Epidemiol Demog & Biometry Program, NIH, Bethesda, MD 20892 USA. Eastern Virginia Med Sch, Strelitz Diabet Inst, Norfolk, VA 23501 USA. Univ Calif San Francisco, Dept Epidemiol & Biostat, San Francisco, CA 94143 USA. Hebrew Rehabil Ctr Aged, Boston, MA 02131 USA. Johns Hopkins Univ, Dept Internal Med, Baltimore, MD USA. Johns Hopkins Univ, Dept Epidemiol, Baltimore, MD USA. RP Resnick, HE (reprint author), Medstar Res Inst, 108 Irving St NW, New York, NY 10010 USA. NR 34 TC 73 Z9 74 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD NOV PY 2000 VL 23 IS 11 BP 1642 EP 1647 DI 10.2337/diacare.23.11.1642 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 368FD UT WOS:000090106300007 PM 11092286 ER PT J AU Giovannone, B Scaldaferri, ML Federici, M Porzio, O Lauro, D Fusco, A Sbraccia, P Borboni, P Lauro, R Sesti, G AF Giovannone, B Scaldaferri, ML Federici, M Porzio, O Lauro, D Fusco, A Sbraccia, P Borboni, P Lauro, R Sesti, G TI Insulin receptor substrate (IRS) transduction system: distinct and overlapping signaling potential SO DIABETES-METABOLISM RESEARCH AND REVIEWS LA English DT Review DE insulin signaling; insulin receptor substrate; IRS-1; IRS-2; IRS-3; IRS-4 ID GROWTH-FACTOR-I; 60-KDA PHOSPHOTYROSINE PROTEIN; RAT ADIPOSE-CELLS; HEMATOPOIETIC-CELLS; GLUT4 TRANSLOCATION; GLUCOSE-TRANSPORT; 3T3-L1 ADIPOCYTES; CARCINOMA-CELLS; MICE; OVEREXPRESSION AB Insulin receptor substrate (IRS) proteins play a central role in maintaining basic cellular functions such as growth and metabolism. They act as an interface between multiple growth factor receptors possessing tyrosine kinase activity, such as the insulin receptor, and a complex network of intracellular signalling molecules containing Src homology 2 (SH2) domains. Four members (IRS-1, IRS-2, IRS-3, IRS-4) of this family have been identified which differ in their subcellular distribution and interaction with SH2 domain proteins. In addition, differential IRS tissue- and developmental-specific expression patterns may contribute to specificity in their signaling potential. Copyright (C) 2000 John Wiley & Sons, Ltd. C1 Univ Roma Tor Vergata, Dipartimento Med Interna, Mol Med Lab, I-00133 Rome, Italy. NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Sesti, G (reprint author), Univ Roma Tor Vergata, Dipartimento Med Interna, Mol Med Lab, Via Tor Vergata 135, I-00133 Rome, Italy. RI Sesti, Giorgio/B-1509-2012; Federici, Massimo/G-9940-2012; OI Federici, Massimo/0000-0003-4989-5194; Lauro, Davide/0000-0002-8597-4415; Sesti, Giorgio/0000-0002-1618-7688 NR 51 TC 94 Z9 102 U1 1 U2 6 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 1520-7552 J9 DIABETES METAB RES JI Diabetes-Metab. Res. Rev. PD NOV-DEC PY 2000 VL 16 IS 6 BP 434 EP 441 DI 10.1002/1520-7560(2000)9999:9999<::AID-DMRR159>3.0.CO;2-8 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 392QV UT WOS:000166423900004 PM 11114102 ER PT J AU Saeki, K Xie, J Notkins, AL AF Saeki, K Xie, J Notkins, AL TI Genomic structure of mouse IA-2: comparison with its human homologue SO DIABETOLOGIA LA English DT Article DE IA-2; insulin-dependent diabetes mellitus; autoantibodies; genomic structure; protein tyrosine phosphatase; intron-exon junctions ID PROTEIN-TYROSINE-PHOSPHATASE; DEPENDENT DIABETES-MELLITUS; TRANSMEMBRANE PROTEIN; INSULIN; IA-2-BETA; GENE; ORGANIZATION; AUTOANTIGEN; SEQUENCE; FAMILY AB Aims/hypothesis. IA-2 is a transmembrane protein with a tyrosine phosphatase (PTP)-like structure and a major autoantigen in Type I (insulin-dependent) diabetes mellitus. Because the nucleotide sequence of human and mouse IA-2 cDNA are closely related, it seemed likely that the genomic organization of the two molecules would be similar. To test this possibility the current experiments were initiated to characterize and compare the genomic structure of mouse and human IA-2. Methods. IA-2 cDNA was used to screen a 129SVJ mouse genomic library. We selected and mapped 7 overlapping clones. The subcloned inserts were used to determine intron-exon junctions by direct sequencing. Polymerase chain reaction and restriction mapping were used to estimate the size of the introns. Results. The mouse IA-2 gene and the 5' upstream regulatory region were isolated and the intron-exon junctions determined. Mouse IA-2 encompasses approximately 20 kb and encodes 23 exons. Both the 3' and 5' ends were mapped by rapid amplification of cDNA ends (RACE) and a 2 kb 5'-upstream region was shown to have functional promoter activity. Conclusion/interpretation. Comparison of the genomic structure of mouse and human IA-2 shows that they have the same number of exons and nearly identical intron-exon junctions. The region around the major transcription start site of mouse IA-2 is similar to human IA-2 and other transmembrane protein tyrosine phosphatases. It is concluded that human and mouse IA-2 are highly conserved and derived from a common ancestral gene. C1 Natl Inst Dent & Craniofacial Res, Expt Med Sect, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Notkins, AL (reprint author), Natl Inst Dent & Craniofacial Res, Expt Med Sect, Oral Infect & Immun Branch, NIH, Bldg 30,Room 121,30 Convent Dr MSC 4322, Bethesda, MD 20892 USA. NR 21 TC 6 Z9 7 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD NOV PY 2000 VL 43 IS 11 BP 1429 EP 1434 DI 10.1007/s001250051550 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 375TD UT WOS:000165416600015 PM 11126414 ER PT J AU Amin, A Finkelstein, R AF Amin, A Finkelstein, R TI Epidermal growth factor receptor signaling activates orthodenticle expression during Drosophila head development SO DNA AND CELL BIOLOGY LA English DT Article ID EMBRYONIC VENTRAL ECTODERM; EGF RECEPTOR; SPITZ GENE; ADULT HEAD; EYE; PATHWAY; WINGLESS; TRANSCRIPTION; HEDGEHOG; PROTEIN AB The relation between signal transduction pathways and the genes that specify regional identity remains poorly understood. We investigated the interaction between the epidermal growth factor receptor (EGFR) pathway and the homeobox gene orthodenticle (otd), which specifies cell fate during head development. Previous studies of head formation in Drosophila melanogaster demonstrated that reducing either EGFR signaling or otd expression in the imaginal primordium of the dorsal head capsule eliminates the ocelli and other dorsal head structures. Here, we show that blocking EGFR signaling reduces otd expression and that activating EGFR signaling outside this primordium induces ectopic otd expression. We also demonstrate that loss of EGFR can be rescued by constitutive otd expression. Our results indicate that otd is a downstream target of the EGFR pathway during head development. C1 NINDS, Bethesda, MD 20892 USA. RP Finkelstein, R (reprint author), NINDS, 6001 Execut Blvd,Room 2142, Bethesda, MD 20892 USA. NR 41 TC 6 Z9 6 U1 1 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1044-5498 J9 DNA CELL BIOL JI DNA Cell Biol. PD NOV PY 2000 VL 19 IS 11 BP 631 EP 638 DI 10.1089/10445490050199036 PG 8 WC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity GA 378CE UT WOS:000165566100001 PM 11098214 ER PT J AU Grant, BF AF Grant, BF TI Theoretical and observed subtypes of DSM-IV alcohol abuse and dependence in a general population sample SO DRUG AND ALCOHOL DEPENDENCE LA English DT Article DE alcohol abuse; alcohol dependence; subtypes of alcohol dependence; subtypes of alcohol abuse; nosology ID UNITED-STATES AB The purpose of this study was to quantify the degree of heterogeneity of the Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-IV) alcohol abuse and dependence categories by comparing the number of theoretically predicted subtypes of each category with those observed in a nationally representative sample of the US general population. Among respondents classified with a past year diagnosis of abuse, only 11 (47.8%) of the 23 theoretically predicted subtypes of abuse were observed, while 53 (53.5%) of the 99 theoretically predicted subtypes of dependence were observed in this general population sample. Approximately 90% of the respondents classified with abuse could be represented by three subtypes of abuse and 70% of the respondents with current diagnoses of dependence could be characterized by six subtypes of dependence, indicating the relative homogeneity of both diagnostic categories. Sociodemographic differentials were also observed including the reduction in the number of observed subtypes of abuse and dependence with age as well as the larger numbers of subtypes associated with males and whites relative to females and blacks, respectively. Implications of these results are discussed in terms of increased physical morbidity and disruption of family life as persons with alcohol use disorders age, the potential role of physiological and impaired control over drinking indicators of dependence as critical features of the disorder, and the future need to examine the conceptual basis of the abuse category and to conduct longitudinal epidemiological research. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 NIAAA, Div Biometry & Epidemiol, Bethesda, MD 20892 USA. RP Grant, BF (reprint author), NIAAA, Div Biometry & Epidemiol, Suite 514,MSC-7003,6000 Execut Blvd, Bethesda, MD 20892 USA. NR 12 TC 28 Z9 28 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0376-8716 J9 DRUG ALCOHOL DEPEN JI Drug Alcohol Depend. PD NOV 1 PY 2000 VL 60 IS 3 BP 287 EP 293 DI 10.1016/S0376-8716(00)00115-0 PG 7 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 368AV UT WOS:000090096100008 PM 11053763 ER PT J AU Hoffmann, C Heine, P Pradel, G Kim, YC Jacobson, KA Zimmermann, H AF Hoffmann, C Heine, P Pradel, G Kim, YC Jacobson, KA Zimmermann, H TI Inhibition of ecto-apyrase and ecto-ATPase by pyridoxal phosphate-related compounds SO DRUG DEVELOPMENT RESEARCH LA English DT Article DE PPADS; enzyme-inhibition; ecto-ATPase; ecto-apyrase; CD39; P2 receptors ID P-2-PURINOCEPTOR ANTAGONISTS; P2-RECEPTOR ANTAGONISTS; NUCLEOTIDASES; BLOCKADE; SUBTYPES; RECEPTORS; DIPHOSPHOHYDROLASE AB Studies of nucleotide receptors (P2-receptors) in cells and tissues are complicated by cleavage of phosphate groups from nucleotide agonist ligands by ecto-nucleotidases. Some P2 receptor antagonists may also inhibit ecto-nucleotidases, making these studies even more complex. In order to systematically approach this problem, we investigated structure-activity relationships of pyridoxal-5'-phosphate-6-azophenyl-2,4-disulfonate (PPADS) and 14 derivatives, many potent as antagonists at P2 receptors, as inhibitors of ecto-nucleotidases. The compounds were tested for their ability to inhibit enzymatic nucleotide breakdown by CHO cells stably transfected with plasmids containing the cDNA for rat ecto-apyrase (NTPDase1) and rat ecto-ATPase (NTPDase2). All inhibitors were tested at a concentration of 100 muM and ATP hydrolysis was quantified by HPLC. Maximal inhibition obtained for ecto-apyrase and ecto-ATPase was 60% and 35%, respectively. Most PPADS analogs were better inhibitors of ecto-apyrase than of ecto-ATPase. Compound 8, a phosphate derivative, inhibited ecto-apyrase with no inhibition evident at ecto-ATPase. Comparison of pharmacological data of PPADS analogs at P2 receptors as previously determined showed that four PPADS analogs exhibited selectivity for P2X nucleotide receptors. None of these compounds inhibited ecto-ATPase, while two inhibited the ecto-apyrase. Compound 14, a bisphosphate derivative, inhibited ecto-ATPase without inhibition of ecto-apyrase. This compound only weakly antagonized P2X(1) receptors and was inactive at P2X(2) and P2Y(1) receptors, thus bearing some selectivity for ecto-ATPase. Compound 7, a 5-methylphosphonate derivative, a potent antagonist of P2X(1) receptors, was inactive at ecto-apyrase and only weakly inhibitory at ecto-ATPase. Thus, PPADS modifications that enhance selectivity among ecto-nucleotidases and P2 receptors have been identified. Drug Dev. Res. 51:153-158, 2000. Published 2001 Wiley-Liss, Inc. C1 NIDDK, Bioorgan Chem Lab, Mol Recognit Sect, NIH, Bethesda, MD 20892 USA. Univ Frankfurt, AK Neurochem, Biozentrum, Frankfurt, Germany. RP Jacobson, KA (reprint author), NIDDK, Bioorgan Chem Lab, Mol Recognit Sect, NIH, Bldg 8A,Rm B1A-19, Bethesda, MD 20892 USA. RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 22 TC 7 Z9 7 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0272-4391 J9 DRUG DEVELOP RES JI Drug Dev. Res. PD NOV PY 2000 VL 51 IS 3 BP 153 EP 158 DI 10.1002/1098-2299(200011)51:3<153::AID-DDR3>3.0.CO;2-X PG 6 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 398PB UT WOS:000166763900003 ER PT J AU Radko, SP Stastna, M Chrambach, A AF Radko, SP Stastna, M Chrambach, A TI Capillary zone electrophoresis of sub-mu m-sized particles in electrolyte solutions of various ionic strengths: Size-dependent electrophoretic migration and separation efficiency SO ELECTROPHORESIS LA English DT Article DE capillary zone electrophoresis; size-dependent separation ID CONCENTRATED COLLOIDAL DISPERSIONS; POLYSTYRENE LATEX; OXIDE PARTICLES; ADSORPTION; SURFACE; CONDUCTIVITY; MOBILITY; MODEL AB To gain insight into the mechanisms of size-dependent separation of microparticles in capillary zone electrophoresis (CZE), sulfated polystyrene latex microspheres of 139, 189, 268, and 381 nm radius were subjected to CZE in Tris-borate buffers of various ionic strengths ranging from 0.0003 to 0.005, at electric field strengths of 100-500 V cm-l. Size-dependent electrophoretic migration of polystyrene particles in CZE was shown to be an explicit function of kappaR, where kappa (-1) and rare the thickness of electric double layer (which can be derived from the ionic strength of the buffer) and particle radius, respectively. Particle mobility depends on kappaR in a manner consistent with that expected from the Overbeek-Booth electrokinetic theory, though a charged hairy layer on the surface of polystyrene latex particles complicates the quantitative prediction and optimization of size-dependent separation of such particles in CZE. However, the Overbeek-Booth theory remains a useful general guide for size-dependent separation of microparticles in CZE. In accordance with it, it could be shown that, for a given pair of polystyrene particles of different sizes, there exists an ionic strength which provides the optimal separation selectivity. Peak spreading was promoted by both an increasing electric field strength and a decreasing ionic strength. When the capillary is efficiently thermostated, the electrophoretic heterogeneity of polystyrene microspheres appears to be the major contributor to peak spreading. Yet, at both elevated electric field strengths (500 V/cm) and the highest ionic strength used (0.005), thermal effects in a capillary appear to contribute significantly to peak spreading or can even dominate it. C1 NICHHD, Macromol Anal Sect, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. Russian Acad Med Sci, Med Genet Res Ctr, Moscow, Russia. RP Chrambach, A (reprint author), NICHHD, Macromol Anal Sect, Lab Cellular & Mol Biophys, NIH, Bldg 10,Rm 9D50, Bethesda, MD 20892 USA. RI Stastna, Miroslava/G-9266-2014 NR 42 TC 49 Z9 52 U1 1 U2 21 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0173-0835 J9 ELECTROPHORESIS JI Electrophoresis PD NOV PY 2000 VL 21 IS 17 BP 3583 EP 3592 DI 10.1002/1522-2683(200011)21:17<3583::AID-ELPS3583>3.0.CO;2-Q PG 10 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 379AV UT WOS:000165619500008 PM 11271475 ER PT J AU Wang, GS Louis, JM Sondej, M Seok, YJ Peterkofsky, A Clore, GM AF Wang, GS Louis, JM Sondej, M Seok, YJ Peterkofsky, A Clore, GM TI Solution structure of the phosphoryl transfer complex between the signal transducing proteins HPr and IIA(Glucose) of the Escherichia coli phosphoenolpyruvate : sugar phosphotransferase system SO EMBO JOURNAL LA English DT Article DE E.coli PEP : sugar PTS; glucose-specific enzyme IIA(Glucose); histidine-containing phosphocarrier protein; NMR ID HISTIDINE-CONTAINING PROTEIN; N-TERMINAL DOMAIN; RESTRAINED MOLECULAR-DYNAMICS; RESIDUAL DIPOLAR COUPLINGS; HIGH-RESOLUTION STRUCTURE; ACTIVE-SITE HISTIDINES; PHOSPHOCARRIER PROTEIN; BACILLUS-SUBTILIS; ENZYME-I; SALMONELLA-TYPHIMURIUM AB The solution structure of the second protein-protein complex of the Escherichia coli phosphoenolpyruvate : sugar phosphotransferase system, that between histidine-containing phosphocarrier protein (HPr) and glucose-specific enzyme IIA(Glucose) (IIA(Glc)), has been determined by NMR spectroscopy, including the use of dipolar couplings to provide long-range orientational information and newly developed rigid body minimization and constrained/restrained simulated annealing methods. A protruding convex surface on HPr interacts with a complementary concave depression on IIA(GlC). Both binding surfaces comprise a central hydrophobic core region surrounded by a ring of polar and charged residues, positive for HPr and negative for IIA(GlC). Formation of the unphosphorylated complex, as well as the phosphorylated transition state, involves little or no change in the protein backbones, but there are conformational rearrangements of the interfacial side chains. Both HPr and IIA(Glc) recognize a variety of structurally diverse proteins. Comparisons with the structures of the enzyme I-HPr and IIA(Glc)-glycerol kinase complexes reveal how similar binding surfaces can be formed with underlying backbone scaffolds that are structurally dissimilar and highlight the role of redundancy and side chain conformational plasticity. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. Seoul Natl Univ, Coll Nat Sci, Dept Microbiol, Seoul 151742, South Korea. RP Clore, GM (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 5, Bethesda, MD 20892 USA. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 83 TC 101 Z9 102 U1 0 U2 11 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 1 PY 2000 VL 19 IS 21 BP 5635 EP 5649 DI 10.1093/emboj/19.21.5635 PG 15 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 372LK UT WOS:000165235000005 PM 11060015 ER PT J AU Tong, WH Rouault, T AF Tong, WH Rouault, T TI Distinct iron-sulfur cluster assembly complexes exist in the cytosol and mitochondria of human cells SO EMBO JOURNAL LA English DT Article DE human; iron; IRP1; iscU; localization ID OXIDATIVE DNA-DAMAGE; SACCHAROMYCES-CEREVISIAE; AZOTOBACTER-VINELANDII; METABOLISM; PROTEINS; YEAST; GENE; GLYCOSYLASES; FERREDOXIN; TRANSPORT AB Iron-sulfur (Fe-S) clusters are cofactors found in many proteins that have important redox, catalytic or regulatory functions. In mammalian cells, almost all known Fe-S proteins are found in the mitochondria, but at least one is found in the cytosol. Here we report cloning of the human homologs to IscU and NifU, iron-binding proteins that play a critical role in Fe-S cluster assembly in bacteria. In human cells, alternative splicing of a common pre-mRNA results in synthesis of two proteins that differ at the N-terminus and localize either to the cytosol (IscU1) or to the mitochondria (IscU2), Biochemical analyses demonstrate that IscU proteins specifically associate with IscS, a cysteine desulfurase that is proposed to sequester inorganic sulfur for Fe-S cluster assembly. Protein complexes containing IscU and IscS can be found in the mitochondria as well as in the cytosol, implying that Fe-S cluster assembly takes place in multiple subcellular compartments in mammalian cells. The possible roles of the IscU proteins in mammalian cells and the potential implications of compartmentalization of Fe-S cluster assembly are discussed. C1 NICHHD, Cell Biol & Metab Branch, Bethesda, MD 20892 USA. RP Rouault, T (reprint author), NICHHD, Cell Biol & Metab Branch, Bethesda, MD 20892 USA. NR 34 TC 128 Z9 133 U1 2 U2 6 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD NOV 1 PY 2000 VL 19 IS 21 BP 5692 EP 5700 DI 10.1093/emboj/19.21.5692 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 372LK UT WOS:000165235000010 PM 11060020 ER PT J AU Whitby, LG Axelrod, J Weil-Malherbe, H AF Whitby, LG Axelrod, J Weil-Malherbe, H TI Endocrine Classic - The fate of H-3-Norepinephrine in animals (Reprinted from The Journal of Pharmacology and Experimental Therapeutics, vol 132, pg 193, 1961) SO ENDOCRINOLOGIST LA English DT Reprint C1 NIMH, Clin Sci Lab, NIH, Bethesda, MD 20892 USA. NIMH, Clin Neuropharmacol Res Ctr, NIH, Bethesda, MD 20892 USA. RP Whitby, LG (reprint author), Univ Cambridge, Dept Biochem, Cambridge CB2 1TN, England. NR 19 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1051-2144 J9 ENDOCRINOLOGIST JI Endocrinologist PD NOV PY 2000 VL 10 IS 6 BP 364 EP 372 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 382XM UT WOS:000165852700003 ER PT J AU Sekar, N Lavoie, HA Veldhuis, JD AF Sekar, N Lavoie, HA Veldhuis, JD TI Concerted regulation of steroidogenic acute regulatory gene expression by luteinizing hormone and insulin (or insulin-like growth factor I) in primary cultures of porcine granulosa-luteal cells SO ENDOCRINOLOGY LA English DT Article ID SIDE-CHAIN CLEAVAGE; FOLLICLE-STIMULATING-HORMONE; LEYDIG TUMOR-CELLS; MESSENGER-RIBONUCLEIC-ACID; PROTEIN STAR GENE; POLYCYSTIC-OVARY-SYNDROME; ELEMENT-BINDING PROTEIN; NUCLEAR RECEPTOR SF-1; C/EBP-BETA; CATALYTIC SUBUNIT AB The steroidogenic acute regulatory (StAR) protein is indispensable for maximal trophic hormone-stimulated steroidogenesis by the adrenal gland, testis, and ovary. Recently, our laboratory developed an in vitro primary culture system of porcine granulosa-luteal cells that retain responsiveness to LH and show LH and insulin [or insulin-like growth factor (IGF-I)] synergy in stimulating StAR messenger RNA accumulation. Here, we examine the mechanisms subserving this LH-insulin (IGF-I) augmentation. We corroborate LH's amplification of insulin as well as IGF-I-stimulated granulosa-luteal cell progesterone and cAMP accumulation (P < 0.001). Insulin or ICF-I elevated LH receptor transcript accumulation, and LH did not alter this effect. To determine the hormonal responsiveness of StAR promoter, truncated regions of the -1423 to +130 bp upstream sequence of the porcine gene were ligated into a firefly luciferase reporter plasmid. Transient transfection of the StAR plasmid containing the full-length porcine 5'-flanking region of StAR (pStAR1423/luc) showed superadditive stimulation by LH and insulin or IGF-I after 24 h. LH, but not insulin or IGF-I alone, stimulated pStAR1423/luc activity. Deletion of the proximal putative steroidogenic factor-1 (-48 to -41) site abolished hormonally driven StAR promoter activity. A stable cAMP analog, 8-bromo-cAMP (1 mM), and insulin/IGF-I also evoked supraadditive StAR promoter expression. To further explore the role of cAMP in LH-insulin (or IGF-I) actions, we cotransfected a Rous sarcoma virus (RSV)-driven minigene encoding the heat-stable inhibitor of the cAMP-dependent protein kinase (RSV/ PKI) or a mutant plasmid (RSV/PKImut) along with the pStAR1423/luc promoter construct. Cotransfection of PKI, but not PKImut, with pStAR1423/luc significantly attenuated LH's stimulation of luciferase activity and also reduced the magnitude of the transcriptional amplification exerted by LH and insulin or IGF-I. In corollary analyses of the protein kinase A (PKA) pathway, cotransfection of full-length pStAR1423/luc and a complementary DNA encoding a constitutively activated PKA catalytic subunit elevated basal and insulin (or IGF-I)-stimulated StAR promoter expression. LH and insulin (or IGF-I) also augmented steady state StAR transcript levels, as assessed by homologous RT-PCR, and StAR protein concentrations, as evaluated by Western blotting. Together, these investigations document a significant role for insulin or IGF-I in enhancing LH-stimulated progesterone and cAMP biosynthesis and endogenous StAR message and protein accumulation and in augmenting cAMP-PKA-dependent transcriptional activation of the exogenous StAR promoter. C1 Univ Virginia, Hlth Sci Ctr, Dept Internal Med, Div Endocrinol & Metab,NIH,Specialized Cooperat C, Charlottesville, VA 22908 USA. Univ S Carolina, Sch Med, Dept Neurosci & Cell Biol, Columbia, SC 29208 USA. RP Veldhuis, JD (reprint author), Univ Virginia, Sch Med, Dept Internal Med, Div Endocrine, Box 202, Charlottesville, VA 22908 USA. EM jdv@virginia.edu NR 62 TC 33 Z9 35 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD NOV PY 2000 VL 141 IS 11 BP 3983 EP 3992 DI 10.1210/en.141.11.3983 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 365VC UT WOS:000089970300009 PM 11089528 ER PT J AU Koshimizu, T Tomic, M Wong, AOL Zivadinovic, D Stojilkovic, SS AF Koshimizu, T Tomic, M Wong, AOL Zivadinovic, D Stojilkovic, SS TI Characterization of purinergic receptors and receptor-channels expressed in anterior pituitary cells SO ENDOCRINOLOGY LA English DT Article ID P2X(2) RECEPTOR; ATP RECEPTORS; EXTRACELLULAR NUCLEOTIDES; HORMONE-RELEASE; CALCIUM; RAT; DESENSITIZATION; ADENOSINE; ACTIVATION; LOCALIZATION AB Purinergic G protein-coupled receptors (P2YR) and ion-conducting receptor-channels (P2XR) are present in the pituitary. However, their identification, expression within pituitary cell subpopulations, and the ability to elevate intracellular Ca2+ concentration ([Ca2+](i)) in response to ATP stimulation were incompletely characterized. Here we show that mixed populations of rat anterior pituitary cells express messenger RNA transcripts for P2Y(2)R, P2X(2a)R, P2X(2b)R, P2X(3)R, P2X(4)R, and P2X(7)R. The transcripts and functional P2Y(2)R were identified in lactotrophs and GH3 cells, but not in somatotrophs and gonadotrophs, and their activation by ATP led to an extracellular Ca2+-independent rise in [Ca2+](i) in about 40% of cells tested. Lactotrophs and GH3 cells, but not somatotrophs, also express transcripts for P2X(7)R, P2X(3)R, and P2X(4)R. Functional P2X(7)R were identified in 74% of lactotrophs, whereas 50% of these cells expressed P2X(3)R and 33% expressed PPX4R. Coexpression of these receptor subtypes in single lactotrophs was frequently observed. Purified somatotrophs expressed transcripts for P2X(2a)R and P2X(2b)R, and functional receptors were identified in somatotrophs and Konadotrophs, but not in lactotrophs. Consistent with the cell-specific expression of transcripts for P2X(2)R and P2X(3)R, the expression of their functional heteromers was not evident in pituitary cells. Receptors differed in their capacities to elevate and sustain Ca2+ influx-dependent rise in [Ca2+](i) during the prolonged ATP stimulation. These results indicate that the purinergic system of anterior pituitary is extremely complex and provides an effective mechanism for generating a cell- and receptor-specific Ca2+ signaling pattern in response to a common agonist. C1 NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Stojilkovic, SS (reprint author), NICHD, Sect Cellular Signaling, ERRB, Bldg 49,Room 6A-36,49 Convent Dr, Bethesda, MD 20892 USA. EM stankos@helix.nih.gov RI Wong, Anderson/A-2735-2010; Tomic, Melanija/C-3371-2016; OI Koshimizu, Taka-aki/0000-0001-5292-7535 NR 29 TC 37 Z9 37 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD NOV PY 2000 VL 141 IS 11 BP 4091 EP 4099 DI 10.1210/en.141.11.4091 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 365VC UT WOS:000089970300021 PM 11089540 ER PT J AU Hauache, OM Hu, JX Ray, K Xie, RY Jacobson, KA Spiegel, AM AF Hauache, OM Hu, JX Ray, K Xie, RY Jacobson, KA Spiegel, AM TI Effects of a calcimimetic compound and naturally activating mutations on the human Ca2+ receptor and on Ca2+ receptor/metabotropic glutamate chimeric receptors SO ENDOCRINOLOGY LA English DT Article ID CALCIUM-SENSING RECEPTOR; CELL-SURFACE EXPRESSION; N-LINKED GLYCOSYLATION; CA2+-SENSING RECEPTOR; SIGNAL-TRANSDUCTION; SPORADIC HYPOPARATHYROIDISM; EXTRACELLULAR DOMAIN; BINDING DOMAIN; LIGAND-BINDING; IDENTIFICATION AB Naturally occurring mutations identified in subjects with autosomal dominant hypocalcemia (ADH) and the calcimimetic compound, R-568, have both been reported to increase Ca2+ sensitivity of the Ca2+ receptor (CaR). To gain insight into their mechanism of action, we studied interactions between four different ADR mutations located in the amino-terminal extracellular domain (ECD) and R-568. We found that R-568 increased the sensitivity of three of the ADH mutant receptors, but the Leu125Pro mutant appeared robe maximally left-shifted in that neither R-568 addition nor combining other ADH mutations with Leu125Pro gave increases in sensitivity comparable to those seen with the three other ADH mutations studied. We also made use of truncation and deletion mutants of the CaR and CaR/metabotropic glutamate receptor type 1 (mGluR1) chimeras to study both the site of action of R-568 and the effect of the Leu125Pro activating mutation. R-568 was effective in receptor constructs containing the seven transmembrane domain (7TM) of the CaR, but not in those containing the mGluR1 7TM. R-568, moreover, imparted Ca2+ responsiveness to CaR constructs lacking all or part of the CaR ECD. The Leu125Pro mutation in contrast conferred no or minimal increase in Ca2+ responsiveness to CaR constructs lacking Dart of the CaR ECD but showed a striking increase in basal activity in the context of chimeras containing an mGluR1 7TM. Our results localize the site of action of NPS-568 specifically to the CaR 7TM. Our results with the Leu125Pro mutant, furthermore, suggest that the mGluR1 7TM domain may be more permissive for activation than the 7TM domain of the CaR. C1 NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Spiegel, AM (reprint author), NIDDK, Metab Dis Branch, NIH, Bldg 31,Room 9A-52, Bethesda, MD 20892 USA. EM spiegela@extra.niddk.nih.gov RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 NR 26 TC 65 Z9 67 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD NOV PY 2000 VL 141 IS 11 BP 4156 EP 4163 DI 10.1210/en.141.11.4156 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 365VC UT WOS:000089970300029 PM 11089548 ER PT J AU Bottner, A Haidan, A Eisenhofer, G Kristensen, K Castle, AL Scherbaum, WA Schneider, H Chrousos, GP Bornstein, SR AF Bottner, A Haidan, A Eisenhofer, G Kristensen, K Castle, AL Scherbaum, WA Schneider, H Chrousos, GP Bornstein, SR TI Increased body fat mass and suppression of circulating leptin levels in response to hypersecretion of epinephrine in phenylethanolamine-N-methyltransferase (PNMT)-overexpressing mice SO ENDOCRINOLOGY LA English DT Article ID SYMPATHETIC-NERVOUS-SYSTEM; TRANSGENIC MICE; ADRENERGIC-RECEPTORS; ADIPOSE-TISSUE; HEART-FAILURE; OBESITY; STRESS; EXPRESSION; LIPOLYSIS; MOUSE AB Epinephrine is a major stress hormone that plays a central role in the control of metabolic function and energy homeostasis. To evaluate the role of epinephrine and the physiological and pathophysiological consequences of sustained elevation of epinephrine on metabolic and endocrine function, we studied several metabolic parameters and circulating leptin levels in a newly developed transgenic mouse model of phenylethanolamine-N-methyltransferase (PNMT) overexpression. A 100-fold overexpression of PNMT and subsequent elevation of epinephrine levels resulted in a marked suppression of circulating leptin levels in the transgenic animals (1.14 +/- 0.05 vs. 2.17 +/- 0.35 ng/ml; P < 0.01), which correlated negatively with plasma epinephrine (r = -0.82; P ( 0.05), thus providing evidence for an inhibitory action of epinephrine on leptin production in vivo. In parallel, we found a marked increase in the body fat content of the transgenic animals (12.54 +/- 1.5 vs. 6.22 +/- 0.2%; P < 0.01) that was accompanied by enlarged adipocytes, indicating an increased lipid storage in PNMT transgenic mice. Interestingly, however, transgenic animals had normal body weight and did not exhibit major alterations in carbohydrate metabolism, as evidenced by analysis of random and fasted blood glucose levels, plasma insulin and C peptide levels, and insulin tolerance test. The metabolic alterations observed were not secondary to changes in food intake or increased activity of the hypothalamic-pituitary-adrenal axis, as there were no differences in these parameters. In summary, sustained primary overproduction of epinephrine resulted in suppression of plasma leptin levels and increased lipid storage in the PNMT transgenic mice. The concerted action of the sympathoadrenal system and reduced leptin may contribute to defending energy reservoirs while maintaining a normal body weight, which may be of vital importance under conditions of stress and energy deficiency. C1 Univ Leipzig, Dept Internal Med 3, D-04103 Leipzig, Germany. NINDS, Intramural Res Program, NIH, Bethesda, MD 20892 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. NICHHD, NIH, Bethesda, MD 20892 USA. Pipeline Biotech AS, Silkeborg, Denmark. Diabet Res Inst, D-40225 Dusseldorf, Germany. Univ Erlangen Nurnberg, Childrens Hosp, D-91054 Erlangen, Germany. RP Bottner, A (reprint author), Univ Leipzig, Dept Internal Med 3, Ph Rosenthal Str 27, D-04103 Leipzig, Germany. EM antje.boettner@gmx.net RI Korner, Antje/B-3988-2015 OI Korner, Antje/0000-0001-6001-0356 NR 46 TC 20 Z9 20 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD NOV PY 2000 VL 141 IS 11 BP 4239 EP 4246 DI 10.1210/en.141.11.4239 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 365VC UT WOS:000089970300039 PM 11089558 ER PT J AU Shalev, A AF Shalev, A TI Discovery of a lipodystrophy gene: one answer, one hundred questions SO EUROPEAN JOURNAL OF ENDOCRINOLOGY LA English DT Article C1 NIDDK, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Shalev, A (reprint author), NIDDK, Diabet Branch, NIH, Bethesda, MD 20892 USA. NR 10 TC 3 Z9 3 U1 0 U2 0 PU BIO SCIENTIFICA LTD PI BRISTOL PA 16 THE COURTYARD, WOODLANDS, BRADLEY STOKE, BRISTOL BS32 4NQ, ENGLAND SN 0804-4643 J9 EUR J ENDOCRINOL JI Eur. J. Endocrinol. PD NOV PY 2000 VL 143 IS 5 BP 565 EP 567 DI 10.1530/eje.0.1430565 PG 3 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 379NG UT WOS:000165648500002 PM 11078977 ER PT J AU Lapointe, R Toso, JF Butts, C Young, HA Hwu, P AF Lapointe, R Toso, JF Butts, C Young, HA Hwu, P TI Human dendritic cells require multiple activation signals for the efficient generation of tumor antigen-specific T lymphocytes SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE dendritic cell; CD40 ligand; tumor immunity; lipopolysaccharide; immunotherapy ID CHEMOKINE RECEPTOR EXPRESSION; INTERFERON-GAMMA; CUTTING EDGE; MATURATION; CYTOKINES; INTERLEUKIN-12; CD40; DIFFERENTIATION; RECRUITMENT; MAINTENANCE AB Dendritic cells (DC) are specialized cells of the immune system responsible for the initiation and regulation of both cellular and humoral responses. DC function is highly dependent on their level of maturation. In this study, we postulated that full DC maturation would require a combination of activating signals. When cultured monocyte-derived DC received stimulation with CD40 ligand (CD40L) and lipopolysaccharide (LPS) together, the IL-12 secretion increased 5-60-fold and the IL-10 secretion increased 5-15-fold when compared with either stimulation alone. In addition, poly I .C, a double-stranded RNA analog that mimics viral infection, also synergized with CD40L to stimulate DC to secrete high levels of IL-12 and IL-10. Flow cytometry revealed an up-regulation in the expression of CD80, CD86 and CD83 following activation with a soluble trimeric form of CD40L (CD40Ls) or LPS. However, no further up-regulation was observed when both CD40Ls and LPS were used together compared with a single stimulatory signal, suggesting that there was no correlation between the expression of these markers and the level of IL-12/IL-10 secretion. Finally, specific cytotoxic T lymphocytes (CTL) were generated using DC pulsed with a modified HLA-A2-restricted peptide epitope derived from the melanoma antigen MART-1. DC activated with a combination of CD40Ls and LPS were more efficient in eliciting MART-specific reactivity compared to DC activated with CD40Ls or LPS alone. These results demonstrate that multiple maturational signals have a positive impact on the ability of DC to secrete IL-12 and IL-10 and more importantly, to generate antigen-specific T lymphocytes. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. Frederick Canc Res & Dev Ctr, Div Basic Sci, Expt Immunol Lab, Frederick, MD USA. RP Hwu, P (reprint author), NCI, Surg Branch, NIH, Bldg 10,Room 2B42,10 Ctr Dr, Bethesda, MD 20892 USA. RI Young, Howard/A-6350-2008 OI Young, Howard/0000-0002-3118-5111 NR 36 TC 77 Z9 80 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD NOV PY 2000 VL 30 IS 11 BP 3291 EP 3298 DI 10.1002/1521-4141(200011)30:11<3291::AID-IMMU3291>3.0.CO;2-2 PG 8 WC Immunology SC Immunology GA 372UU UT WOS:000165253100026 PM 11093145 ER PT J AU Nakajima, H Cella, M Bouchon, A Grierson, HL Lewis, J Duckett, CS Cohen, JI Colonna, M AF Nakajima, H Cella, M Bouchon, A Grierson, HL Lewis, J Duckett, CS Cohen, JI Colonna, M TI Patients with X-linked lymphoproliferative disease have a defect in 2B4 receptor-mediated NK cell cytotoxicity SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE human; NK; X-linked lymphoproliferative; Epstein-Barr virus; SLAM-associated protein ID NATURAL-KILLER-CELLS; EPSTEIN-BARR-VIRUS; PROTEIN-TYROSINE KINASE; T-CELLS; ENCODING GENE; CUTTING EDGE; ACTIVATION; MOLECULE; CD48; SUPERFAMILY AB Patients with the X-linked lymphoproliferative disorder (XLPD) are unable to control Epstein-Barr virus (EBV)-induced infections and lymphoproliferation. This disease is caused by a deficit of SAP, an adapter protein involved in the signal transduction of several cell surface receptors of the CD2 superfamily. One of these receptors, called 2B4, is expressed on NK cells, cytotoxic T cells and myeloid cells and activates NK cell cytotoxicity. Here we show that XLPD patients have a defect of 2B4 receptor-mediated NK cell cytotoxicity. This defect may contribute to the pathogenesis of XLPD by reducing NK cell lysis of EBV-infected B cells. C1 Basel Inst Immunol, CH-4005 Basel, Switzerland. NIH, Med Virol Sect, Clin Invest Lab, Bethesda, MD 20892 USA. NIH, Metab Branch, Bethesda, MD 20892 USA. RP Colonna, M (reprint author), Basel Inst Immunol, Grenzacherstr 487, CH-4005 Basel, Switzerland. RI Cella, Marina/G-8850-2011; OI Colonna, Marco/0000-0001-5222-4987 NR 43 TC 130 Z9 132 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD NOV PY 2000 VL 30 IS 11 BP 3309 EP 3318 DI 10.1002/1521-4141(200011)30:11<3309::AID-IMMU3309>3.0.CO;2-3 PG 10 WC Immunology SC Immunology GA 372UU UT WOS:000165253100028 PM 11093147 ER PT J AU Rossi, J Tomac, A Saarma, M Airaksinen, MS AF Rossi, J Tomac, A Saarma, M Airaksinen, MS TI Distinct roles for GFR alpha 1 and GFR alpha 2 signalling in different cranial parasympathetic ganglia in vivo SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE GDNF; lacrimal; mouse; neurturin; otic ganglia; Ret; salivary; sphenopalatine; submandibular ID SUBMANDIBULAR GANGLION; NEURONS; NEURTURIN; RECEPTOR; GDNF; EXPRESSION; GROWTH; TARGET; MOUSE; RET AB Neurturin (NRTN), signalling via the GDNF family receptor alpha2 (GFR alpha2) and Ret tyrosine kinase, has recently been identified as an essential target-derived factor for many parasympathetic neurons. NRTN is expressed in salivary and lacrimal glands, while GFR alpha2 and Ret are expressed in the corresponding submandibular, otic and sphenopalatine ganglia. Here, we have characterized in more detail the role of GDNF and NRTN signalling in the development of cranial parasympathetic neurons and their target innervation. Gfra1 mRNA was expressed at E12 but not in newborn cranial parasympathetic ganglia, while Gfra2 mRNA and protein were strongly expressed in newborn and adult cranial parasympathetic neurons and their projections, respectively. In newborn GFR alpha1- or Ret-deficient mice, where many submandibular ganglion neurons were still present, the otic and sphenopalatine ganglia were completely missing. In contrast, in newborn GFR alpha2-deficient mice, most neurons in all these ganglia were present. In these mice, the loss and atrophy of the submandibular and otic neurons were amplified postnatally, accompanied by complete loss of innervation in some target regions and preservation in others. Surprisingly, GFR alpha2-deficient sphenopalatine neurons, whose targets were completely uninnervated, were not reduced in number and only slightly atrophied. Thus, GDNF signalling via GFR alpha1/Ret is essential in the early gangliogenesis of some, but not all, cranial parasympathetic neurons, whereas NRTN signalling through GFR alpha2/Ret is essential for the development and maintenance of parasympathetic target innervation. These results indicate that GDNF and NRTN have distinct functions in developing parasympathetic neurons, and suggest heterogeneity among and within different parasympathetic ganglia. C1 Viikki Bioctr, Inst Biotechnol, Program Mol Neurobiol, FIN-00014 Helsinki, Finland. NIH, Lab Mammalian Genes & Dev, Bethesda, MD 20892 USA. RP Airaksinen, MS (reprint author), Viikki Bioctr, Inst Biotechnol, Program Mol Neurobiol, FIN-00014 Helsinki, Finland. OI Rossi, Jari/0000-0002-3728-9800; Airaksinen, Matti/0000-0002-7199-4984 NR 23 TC 30 Z9 30 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD NOV PY 2000 VL 12 IS 11 BP 3944 EP 3952 DI 10.1046/j.1460-9568.2000.00292.x PG 9 WC Neurosciences SC Neurosciences & Neurology GA 378XD UT WOS:000165609900014 PM 11069590 ER PT J AU Stafford, D Rice, KC Lewis, DB Glowa, JR AF Stafford, D Rice, KC Lewis, DB Glowa, JR TI Response requirements and unit dose modify the effects of GBR 12909 on cocaine-maintained behavior SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Article ID ALTERNATIVE NONDRUG REINFORCER; FREELY MOVING RATS; PRICE ANALYSIS; PHARMACOLOGICAL TREATMENTS; NUCLEUS-ACCUMBENS; RHESUS-MONKEYS; DRUG; DOPAMINE; ECONOMICS; FOOD AB Previous studies found that GBR 12909 can decrease cocaine-maintained responding at doses that do not affect food-maintained responding. In this study, the effects of GBR 12909 (0.3-3.0 mg/kg) were further examined by varying the response requirement and unit dose of cocaine. Rhesus monkeys earned food or cocaine under a multiple fixed-ratio (FR) schedule. The FR for food was always 30, but the FR for cocaine was varied from 10-130 and the unit dose was varied from 5.6-56.0 mug/kg per injection. Doses of GBR 12909 were tested in an ascending order, for 5 consecutive sessions each. GBR 12909 selectively decreased cocaine-maintained responding in all monkeys in at least 1 condition. These effects were enhanced with large response requirements and/or small unit doses. The results demonstrate that environmental variables can influence the selectivity of GBR 12909's effects and contribute to a growing debate concerning the evaluation of potential pharmacotherapies for drug abuse. C1 Louisiana State Univ, Med Ctr, Dept Pharmacol & Expt Therapeut, Shreveport, LA 71130 USA. NIDDKD, Med Chem Lab, Bethesda, MD 20892 USA. RP Glowa, JR (reprint author), Louisiana State Univ, Med Ctr, Dept Pharmacol & Expt Therapeut, 1501 Kings Highway, Shreveport, LA 71130 USA. FU NIDA NIH HHS [DA 09820] NR 40 TC 11 Z9 11 U1 5 U2 5 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD NOV PY 2000 VL 8 IS 4 BP 539 EP 548 DI 10.1037//1064-1297.8.4.539 PG 10 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 379BE UT WOS:000165620400009 PM 11127425 ER PT J AU Weingartner, H AF Weingartner, H TI Metaphors and models of executive functioning: Comment on Giancola (2000) SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Editorial Material ID COGNITIVE-DEVELOPMENT; PREFRONTAL CORTEX; TRIAZOLAM; MEMORY; CONSTRUCTION AB Clinical researchers have become increasingly interested in considering the role of normal and impaired executive functioning in psychopathology. The concept of executive functions is, however, often used as a metaphor for a wide range of operations that may, on the one hand, be distinct from one another, but also may be integrated in various ways under different information-processing conditions. A clinician's perspective of executive functioning should rake full advantage of the rich body of data and theory that has developed in contemporary cognitive neuroscience around functions such as forms of inhibition and controlled (in contrast to automatic) functions, in a variety of cognitive domains such as making decisions and tracking performance. C1 Natl Inst Drug Abuse, Div Basic Res, Bethesda, MD 20892 USA. RP Weingartner, H (reprint author), Natl Inst Drug Abuse, Div Basic Res, 6001 Execut Blvd,Room 4282,MSC 9555, Bethesda, MD 20892 USA. NR 17 TC 4 Z9 4 U1 2 U2 2 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD NOV PY 2000 VL 8 IS 4 BP 609 EP 611 DI 10.1037/1064-1297.8.4.609 PG 3 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 379BE UT WOS:000165620400018 PM 11127434 ER PT J AU Rialas, CM Nomizu, M Patterson, M Kleinman, HK Weston, CA Weeks, BS AF Rialas, CM Nomizu, M Patterson, M Kleinman, HK Weston, CA Weeks, BS TI Nitric oxide mediates laminin-induced neurite outgrowth in PC12 cells SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE laminin; nitric oxide; PC12 cells; neurite outgrowth; 1-NAME; d-NAME ID PROTEIN-KINASE-C; PERIPHERAL-NERVE REGENERATION; NEURONAL NO SYNTHASE; TYROSINE PHOSPHORYLATION; SYNTHETIC PEPTIDES; HEPARAN-SULFATE; BINDING PROTEIN; RAT-BRAIN; DEPENDENT MECHANISM; CEREBRAL-CORTEX AB Laminin is a potent stimulator of neurite outgrowth in a variety of primary neurons and neuronal cell lines. Here, we investigate the role of nitric oxide in the signaling mechanism of laminin-mediated neurite outgrowth in the PC12 cell line. Within 8 s of exposure to laminin, PC12 cells produce nitric oxide. Peak laminin-induced nitric oxide levels reach 8 nM within 12 s of exposure to laminin and constitutive nitric oxide production is sustained for 1 min. A neurite outgrowth promoting synthetic peptide (AG73), derived from the laminin-l-alpha globular domain, also stimulated nitric oxide release. The nitric oxide synthase inhibitor, 1-NAME, prevents the formation of nitric oxide and here, 1-NAME inhibited both laminin-mediated and AG73-mediated neurite outgrowth by 88 and 95%, respectively. In contrast, C16, a synthetic peptide derived from the laminin-1-gamma chain, is shown here to promote PC12 cell attachment, but not neurite outgrowth. Interestingly, the C16 peptide did not activate nitric oxide release, suggesting that laminin-induced nitric oxide release in PC12 cells is associated only with neurite outgrowth promoting laminin domains and signals. In addition, the data here show that the nitric oxide released by PC12 cells in response to laminin is required as a part of the mechanism of laminin-mediated neurite outgrowth. (C) 2000 Academic Press. C1 Adelphi Univ, Dept Biol, Div Math & Sci, Garden City, NY 11530 USA. SUNY Coll Old Westbury, Neurosci Res Inst, Old Westbury, NY 11568 USA. Hokkaido Univ, Grad Sch Environm Earth Sci, Sapporo, Hokkaido 0600810, Japan. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. SUNY Stony Brook, Dept Pharmacol, Stony Brook, NY 11794 USA. RP Weeks, BS (reprint author), Adelphi Univ, Dept Biol, Div Math & Sci, 1 South Ave, Garden City, NY 11530 USA. NR 64 TC 24 Z9 24 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD NOV 1 PY 2000 VL 260 IS 2 BP 268 EP 276 DI 10.1006/excr.2000.5017 PG 9 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 371XZ UT WOS:000165205700010 PM 11035921 ER PT J AU Curtis, DJ Jane, SM Hilton, DJ Dougherty, L Bodine, DM Begley, CG AF Curtis, DJ Jane, SM Hilton, DJ Dougherty, L Bodine, DM Begley, CG TI Adaptor protein SKAP55R is associated with myeloid differentiation and growth arrest SO EXPERIMENTAL HEMATOLOGY LA English DT Article DE SRC kinase substrate; signaling; hematopoiesis; tyrosine phosphorylation ID LYN-DEFICIENT MICE; RECEPTOR ALPHA-CHAIN; TYROSINE KINASES; THYMOCYTE DEVELOPMENT; TRANSCRIPTION FACTOR; SIGNAL-TRANSDUCTION; ENFORCED EXPRESSION; AUTOIMMUNE-DISEASE; MOLECULAR-CLONING; PROFOUND BLOCK AB Objective. Activation of the SRC family of protein tyrosine kinases is an important component of intracellular signaling in hematopoiesis, but their critical substrates are less well understood. In this report, we describe the cloning and functional characterization of murine SKAP55R (mSKAP55R), an SRC family kinase substrate, Materials and Methods. Expression of mSKAP55R was examined by Northern blot. Phosphorylation of mSKAP55R was examined by transient transfection of COS cells. For overexpression studies, mSKAP55R was cloned into a bicistronic murine stem cell virus-based retrovirus. Transduced cells (FDC-P1 cell line and murine bone marrow) were FACS isolated by expression of the selectable marker green fluorescent protein. Results. mSKAP55R showed 90% amino acid identity to the recently published human SKAP55R. mSKAP55R contained a central pleckstrin homology domain, a C-terminal SH3 domain, and a putative SRC kinase consensus substrate DEIY260. mSKAP55R was expressed in all hematopoietic lineages, with relative mRNA levels greatest in cells of the myeloid and erythroid lineages. Induced myeloid differentiation of M1 and HL-60 cell lines was associated with an eight-fold increase in mSKAP55R mRNA. Transient expression of mSKAP55R in COS cells demonstrated that tyrosine 260 was the predominant site of phosphorylation by FYN kinase, Furthermore, this phosphotyrosine was essential for coimmunoprecipitation of FYN with mSKAP55R. Enforced expression of mSKAP55R inhibited in vitro growth of the myeloid FDC-P1 cell line and primary hematopoietic progenitors, In contrast, a tyrosine 260 mutant mSKAP55R had no effect on in vitro growth. Conclusion. These studies implicate mSKAP55R in the processes of myeloid differentiation and growth arrest. (C) 2000 International Society for Experimental Hematology. Published by Elsevier Science Inc. C1 NHGRI, Hematopoiesis Sect, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. Royal Melbourne Hosp, Rotary Bone Marrow Res Lab, Melbourne, Vic, Australia. Royal Melbourne Hosp, Cooperat Res Ctr Cellular Growth Factors, Melbourne, Vic, Australia. Royal Melbourne Hosp, Walter & Eliza Hall Inst Med Res, Melbourne, Vic, Australia. Western Australian Inst Med Res, Perth, WA, Australia. RP NHGRI, Hematopoiesis Sect, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. EM dcurtis@nhgri.nih.gov RI Jane, Stephen/D-6659-2011; Hilton, Douglas/C-7250-2013 OI Hilton, Douglas/0000-0002-7698-2392 FU PHS HHS [C-22556] NR 47 TC 20 Z9 21 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0301-472X EI 1873-2399 J9 EXP HEMATOL JI Exp. Hematol. PD NOV PY 2000 VL 28 IS 11 BP 1250 EP 1259 DI 10.1016/S0301-472X(00)00537-3 PG 10 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 374RQ UT WOS:000165358500007 PM 11063873 ER PT J AU Guo, ZH Mattson, MP AF Guo, ZH Mattson, MP TI In vivo 2-deoxyglucose administration preserves glucose and glutamate transport and mitochondrial function in cortical synaptic terminals after exposure to amyloid beta-peptide and iron: Evidence for a stress response SO EXPERIMENTAL NEUROLOGY LA English DT Article DE Alzheimer's disease; amyloid; apoptosis; calorie restriction; cerebral cortex; free radicals; heat-shock protein; mitochondria ID PROTECTS HIPPOCAMPAL-NEURONS; MANGANESE SUPEROXIDE-DISMUTASE; MEMBRANE LIPID-PEROXIDATION; FOCAL CEREBRAL-ISCHEMIA; DIETARY RESTRICTION; ALZHEIMERS-DISEASE; OXIDATIVE STRESS; HEAT-SHOCK; PEROXYNITRITE PRODUCTION; CALCIUM HOMEOSTASIS AB Mild metabolic stress can increase resistance of neurons in the brain to subsequent more severe insults, as exemplified by the beneficial effects of heat shock and ischemic preconditioning. Studies of Alzheimer's disease and other age-related neurodegenerative disorders indicate that dysfunction and degeneration of synapses occur early in the cell death process, and that oxidative stress and mitochondrial dysfunction are central events in this pathological process. It was recently shown that administration of 2-deoxy-D-glucose (2DG), a nonmetabolizable glucose analog that induces metabolic stress, to rats and mice can increase resistance of neurons in the brain to excitotoxic, ischemic, and oxidative injury. We now report that administration of 2DG to adult rats (daily i.p. injections of 100 mg/kg body weight) increases resistance of synaptic terminals to dysfunction and degeneration induced by amyloid beta -peptide and ferrous iron, an oxidative insult. The magnitude of impairment of glucose and glutamate transport induced by amyloid beta -peptide and iron was significantly reduced in cortical synaptosomes from 2DG-treated rats compared to saline-treated control rats. Mitochondrial dysfunction, as indicated by increased levels of reactive oxygen species and decreased transmembrane potential, was significantly attenuated after exposure to amyloid beta -peptide and iron in synaptosomes from 2DG-treated rats. Levels of the stress proteins HSP-70 and GRP-78 were increased in synaptosomes from 2DG-treated rats, suggesting a mechanism whereby 2DG protects synaptic terminals. We conclude that 2DG bolsters cytoprotective mechanisms within synaptic terminals, suggesting novel preventative and therapeutic approaches for neurodegenerative disorders. (C) 2000 Academic Press. C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. Univ Kentucky, Sanders Brown Ctr Aging, Lexington, KY 40536 USA. RP Guo, ZH (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 55 TC 53 Z9 60 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4886 J9 EXP NEUROL JI Exp. Neurol. PD NOV PY 2000 VL 166 IS 1 BP 173 EP 179 DI 10.1006/exnr.2000.7497 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 370JM UT WOS:000165119800015 PM 11031093 ER PT J AU Cho-Chung, YS AF Cho-Chung, YS TI Antisense and therapeutic oligonucleotides: toward a gene-targeting cancer clinic SO EXPERT OPINION ON THERAPEUTIC PATENTS LA English DT Review DE antisense; cancer; chemotherapy; DNA methylase; oligonucleotides; oncogenes; protein kinase; transcription factor decoy ID PROTEIN-KINASE-A; ENDOTHELIAL GROWTH-FACTOR; RI-ALPHA-SUBUNIT; VASCULAR-PERMEABILITY FACTOR; TRANSCRIPTION FACTOR DECOY; NF-KAPPA-B; STRAND-BREAK REPAIR; TUMOR-CELL LINES; C-MYC PROTEIN; IN-VIVO AB Dramatic progress is being made in the identification of genes that are involved in oncogenesis and cancer progression. Novel therapeutic oligonucleotides, such as those that can block the expression of specific target genes (antisense) and those that can interfere with a specific enhancer element-directed transcription in vivo (triple helix-forming or decoy), are potentially powerful tools for the therapeutic manipulation of genes associated with cancer. Such a gene-targeting approach is no longer a wishful dream, but a reality that will drive clinical medicine in the foreseeable future. C1 NCI, Cellular Biochem Sect, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. RP Cho-Chung, YS (reprint author), NCI, Cellular Biochem Sect, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. NR 164 TC 11 Z9 12 U1 0 U2 0 PU ASHLEY PUBLICATIONS LTD PI LONDON PA UNITEC HOUSE, 3RD FL, 2 ALBERT PLACE FINCHLEY CENTRAL, LONDON N3 1QB, ENGLAND SN 1354-3776 J9 EXPERT OPIN THER PAT JI Expert Opin. Ther. Patents PD NOV PY 2000 VL 10 IS 11 BP 1711 EP 1724 DI 10.1517/13543776.10.11.1711 PG 14 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 371FG UT WOS:000165167400005 ER PT J AU Andoh, T Lee, SY Chiueh, CC AF Andoh, T Lee, SY Chiueh, CC TI Preconditioning regulation of bcl-2 and p66(shc) by human NOS1 enhances tolerance to oxidative stress SO FASEB JOURNAL LA English DT Article C1 NIMH, Ctr Clin, NIH, Bethesda, MD 20892 USA. RP Chiueh, CC (reprint author), NIMH, Ctr Clin, NIH, NIH Bldg 10,Room 3D-41, Bethesda, MD 20892 USA. NR 0 TC 67 Z9 67 U1 0 U2 1 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD NOV PY 2000 VL 14 IS 14 BP 2144 EP 2146 PG 3 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 371QZ UT WOS:000165190800006 PM 11023998 ER PT J AU Watanabe, A Fujii, I Tsai, HF Chang, YC Kwon-Chung, KJ Ebizuka, Y AF Watanabe, A Fujii, I Tsai, HF Chang, YC Kwon-Chung, KJ Ebizuka, Y TI Aspergillus fumigatus alb1 encodes naphthopyrone synthase when expressed in Aspergillus oryzae SO FEMS MICROBIOLOGY LETTERS LA English DT Article DE melanin; biosynthesis; polyketide synthase; naphthopyrone; tetrahydroxynaphthalene ID NIDULANS WA-GENE; POLYKETIDE SYNTHASE; MELANIN BIOSYNTHESIS; MOLECULAR CHARACTERIZATION; HETEROLOGOUS EXPRESSION; INVASIVE ASPERGILLOSIS; PRODUCT IDENTIFICATION; FUNGI; TRICYCLAZOLE; MODULATION AB Conidial pigment biosynthesis is an important virulence factor in Aspergillus fumigatus, a human fungal pathogen. Involvement of DHN-melanin pathway in the biosynthesis of A. fumigatus conidial pigment implies that the Alb1p polyketide synthase (PKS) is a 1,3,6,8-tetrahydroxynaphthalene (T4HN) synthase. The Alb1p, however, shows higher sequence similarity to a naphthopyrone synthase than to a T4HN synthase. To clarify the function of Alb1p, the alb1 gene was overexpressed in a heterologous host Aspergillus oryzae. The Alb1p PKS product in this heterologous expression system was identified as heptaketide naphthopyrone instead of pentaketide T4HN. The data suggest that Alb1p is a naphthopyrone synthase. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. C1 Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan. NIAID, Clin Invest Lab, Bethesda, MD 20892 USA. RP Ebizuka, Y (reprint author), Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan. NR 28 TC 46 Z9 49 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1097 J9 FEMS MICROBIOL LETT JI FEMS Microbiol. Lett. PD NOV 1 PY 2000 VL 192 IS 1 BP 39 EP 44 DI 10.1111/j.1574-6968.2000.tb09356.x PG 6 WC Microbiology SC Microbiology GA 367DV UT WOS:000090047200007 PM 11040426 ER PT J AU Howell, C Jeffers, L Hoofnagle, JH AF Howell, C Jeffers, L Hoofnagle, JH TI Hepatitis c in African Americans: Summary of a workshop SO GASTROENTEROLOGY LA English DT Review ID DYNAMICS IN-VIVO; TERM FOLLOW-UP; HEPATOCELLULAR-CARCINOMA; UNITED-STATES; CLINICAL OUTCOMES; VIRUS-INFECTION; VIRAL-HEPATITIS; INTERFERON-ALFA; LIVER-DISEASE; PREVALENCE C1 Univ Maryland, Sch Med, Div Gastroenterol & Hepatol, Baltimore, MD 21201 USA. Univ Miami, Sch Med, Ctr Liver Dis, Miami, FL USA. Vet Adm Med Ctr, Miami, FL 33125 USA. NIDDKD, Div Digest Dis & Nutr, NIH, Bethesda, MD 20892 USA. RP Howell, C (reprint author), Univ Maryland, Sch Med, Div Gastroenterol & Hepatol, 22 Green St,N3W130, Baltimore, MD 21201 USA. NR 40 TC 90 Z9 90 U1 1 U2 3 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD NOV PY 2000 VL 119 IS 5 BP 1385 EP + DI 10.1053/gast.2000.19582 PG 13 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 367KU UT WOS:000090060900028 PM 11054398 ER PT J AU Havlik, RJ Guralnik, J AF Havlik, RJ Guralnik, J TI Antihypertensive drugs and gastrointestinal hemorrhage: Causation or confounding SO GASTROENTEROLOGY LA English DT Letter ID RISK C1 NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. Social & Sci Syst, Bethesda, MD USA. Wake Forest Univ, Sch Med, Sticht Ctr Aging, Winston Salem, NC 27109 USA. Duke Univ, Ctr Med, Ctr Study Aging & Human Dev, Durham, NC 27706 USA. RP Havlik, RJ (reprint author), NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. NR 6 TC 0 Z9 0 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD NOV PY 2000 VL 119 IS 5 BP 1416 EP 1417 PG 2 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 367KU UT WOS:000090060900038 PM 11185465 ER PT J AU Vinores, SA Seo, MS Okamoto, N Ash, JD Wawrousek, EF Xiao, WH Hudish, T Derevjanik, NL Campochiaro, PA AF Vinores, SA Seo, MS Okamoto, N Ash, JD Wawrousek, EF Xiao, WH Hudish, T Derevjanik, NL Campochiaro, PA TI Experimental models of growth factor-mediated angiogenesis and blood-retinal barrier breakdown SO GENERAL PHARMACOLOGY-THE VASCULAR SYSTEM LA English DT Article; Proceedings Paper CT 6th Biannual Meeting on Angiogenesis - Basic Science and Clinical Developments CY JUN, 2001 CL IRAKLION, GREECE DE angiogenesis; blood-retinal barrier; vascular endothelial growth factor; platelet-derived growth factor; retinal neovascularization; interleukin-1beta; transgenic mice ID VASCULAR-PERMEABILITY FACTOR; PROLIFERATIVE DIABETIC-RETINOPATHY; CHOROIDAL NEOVASCULAR MEMBRANES; HUMAN GLIOMA-CELLS; FACTOR VEGF; IN-VIVO; ENDOTHELIAL-CELLS; UP-REGULATION; PDGF-BB; FACTOR EXPRESSION AB Following chronic ischemia, vascular endothelial growth factor (VEGF) is induced primarily in the ganglion cell layer of the retina. This often results in neovascularization (NV) that originates from the vascular bed closest to the ganglion cell layer. To study the effects of VEGF, independent lines of transgenic mice that express VEGF in the lens and in the retina have been generated. Expression in the lens results in excessive proliferation and accumulation of angioblasts and endothelial cells in proximity to the lens. However, VEGF expression is not sufficient to direct blood vessel organization or maturation in the prenatal mouse, Abnormal vessels do form on the retinal surface, but not until the second postnatal week. In transgenic mice expressing VEGF in the photoreceptors, NV originates from the deep capillary bed-the vascular bed closest to the photoreceptors. NV is accompanied by localized blood-retinal barrier breakdown. NV is also induced in PDGF-B transgenic mice. PDGF-B expression in the lens occurs prenatally and, during this time, mainly affects the perilenticular vessels. Postnatally, transgenic mice expressing PDGF-B in the lens or photoreceptors show a similar phenotype. In both models, a highly vascularized cell mass containing endothelial cells, pericytes, and glia forms in the superficial retina, and the formation of the deep capillary bed is inhibited. The phenotype suggests that an additional factor is necessary for the maturation and penetration of vascular endothelial cells into the retina to form the deep capillary bed. (C) 2002 Elsevier Science Inc. All rights reserved. C1 Johns Hopkins Univ, Sch Med, Wilmer Eye Inst, Baltimore, MD 21287 USA. Chonnam Natl Univ, Med Sch & Hosp, Dept Ophthalmol, Kwangju, South Korea. Kitasato Univ, Sch Med, Dept Ophthalmol, Kanagawa, Japan. Univ Oklahoma, Hlth Sci Ctr, Dean A McGee Eye Inst, Oklahoma City, OK USA. NEI, NIH, Bethesda, MD 20892 USA. RP Vinores, SA (reprint author), Johns Hopkins Univ, Sch Med, Wilmer Eye Inst, 825 Maumenee Bldg,600 N Wolfe St, Baltimore, MD 21287 USA. EM svinores@jhmi.edu RI Wawrousek, Eric/A-4547-2008 FU NEI NIH HHS [EY10017, EY12190, EY05951] NR 73 TC 8 Z9 8 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0306-3623 J9 GEN PHARMACOL-VASC S JI Gen. Pharmacol.-The Vasc. Syst. PD NOV PY 2000 VL 35 IS 5 BP 233 EP 239 AR PII S0306-3623(01)00117-3 DI 10.1016/S0306-3623(01)00117-3 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 545YY UT WOS:000175246200003 PM 11888678 ER PT J AU Gemma, A Seike, M Seike, Y Uematsu, K Hibino, S Kurimoto, F Yoshimura, A Shibuya, M Harris, CC Kudoh, S AF Gemma, A Seike, M Seike, Y Uematsu, K Hibino, S Kurimoto, F Yoshimura, A Shibuya, M Harris, CC Kudoh, S TI Somatic mutation of the hBUB1 mitotic checkpoint gene in primary lung cancer SO GENES CHROMOSOMES & CANCER LA English DT Article ID STAGE-I ADENOCARCINOMA; DNA CONTENT; CELL-CYCLE; SURVIVAL; KINASE; LINES; BUB1 AB Mutations in mitotic checkpoint genes have been detected in several human cancers, and these cancers exhibit chromosomal instability. Aneuploid stem cells seem to result from chromosomal instability and have been reported in many lung cancers. To determine whether alteration of mitotic checkpoint regulators is involved in carcinogenesis and tumor progression in primary lung cancer, we screened the genomic DNA sequence of 30 human lung cancer cell lines and 30 primary lung cancer tumors for a mutation in the hBUB1 mitotic checkpoint gene. First, we designed 26 sets of intron-based primers to amplify each of the 25 exons of the hBUB1 gene to examine the entire coding region of the hBUB1 gene. Using these primers, we performed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis as well as direct sequencing in the mutation analysis of the hBUB1 gene. Three different nucleotide substitutions were detected in the coding region of the hBUB1 gene in some of the cancer cell lines and primary tumors as follows. The hBUB1 gene of one adenocarcinoma tumor contained a somatic missense mutation, a cytosine-to-guanine substitution in codon 51 of exon 5 that resulted in a histidine-to-aspartic acid amino acid substitution. The hBUB1 gene of three lung cancer cell lines contained a thymine-to-cytosine substitution in codon 430 of exon 12, which did not result in an amino-acid substitution. We were unable to determine whether the nucleotide substitution in exon 12 was a polymorphism or a silent mutation because matched normal tissue was not available. A polymorphism in codon 93 of exon 4, a guanine-to-thymine substitution, in hBUB1 was found in one lung cancer cell line and one primary lung tumor. This is the first report of a somatic missense mutation of a gene involved in a mitotic checkpoint in primary lung cancer. The presence of a point mutation in the hBUB1 gene is consistent with the hypothesis that alteration of mitotic checkpoint genes is involved in the development of primary lung cancers. Because the frequency of hBUB1 gene mutations was low, future studies should focus on other mechanisms of inactivation of the hBUB1 gene as well as mutation analysis of other mitotic checkpoint genes in lung cancers. (C) 2000 Wiley-Liss, Inc. C1 Nippon Med Sch, Dept Internal Med 4, Bunkyo Ku, Tokyo 113, Japan. NCI, Human Carcinogenesis Lab, Bethesda, MD 20892 USA. RP Gemma, A (reprint author), Nippon Med Sch, Dept Internal Med 4, Bunkyo Ku, 1-1-5 Sendagi, Tokyo 113, Japan. NR 27 TC 67 Z9 71 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 1045-2257 J9 GENE CHROMOSOME CANC JI Gene Chromosomes Cancer PD NOV PY 2000 VL 29 IS 3 BP 213 EP 218 DI 10.1002/1098-2264(2000)9999:9999<::AID-GCC1027>3.0.CO;2-G PG 6 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 362YG UT WOS:000089805400003 PM 10992296 ER PT J AU Therrien, M Morrison, DK Wong, AM Rubin, GM AF Therrien, M Morrison, DK Wong, AM Rubin, GM TI A genetic screen for modifiers of a kinase suppressor of ras-dependent rough eye phenotype in drosophila SO GENETICS LA English DT Article ID PROTEIN-KINASE; SIGNAL-TRANSDUCTION; MAP KINASE; TYROSINE KINASE; KSR INTERACTS; C-ELEGANS; MUTATIONS; SEVENLESS; PATHWAY; MEK AB kinase suppressor of Ras (ksr) encodes a putative protein kinase that by genetic criteria appears to function downstream of RAS in multiple receptor tyrosine kinase (RTK) pathways. While biochemical evidence suggests that the role of KSR is closely linked to the signal transduction mechanism of die MAPK cascade, the precise molecular function of KSR remains unresolved. To further elucidate the role of KSR and to identify proteins that may be required for KSR function, we conducted a dominant modifier screen in Drosophila based on a KSR-dependent phenotype. Overexpression of the KSR kinase domain in a subset of cells during Drosophila eye development blocks photoreceptor cell differentiation and results in the external roughening of the adult eye. Therefore, mutations in genes functioning with KSR might modify the KSR-dependent phenotype. We screened similar to 185,000 mutagenized progeny for dominant modifiers of the KSR-dependent rough eye phenotype. A total of 15 complementation groups of Enhancers and four complementation groups of Suppressors were derived. Ten of these complementation groups correspond to mutations in known components of the Ras1 pathway, demonstrating the ability of the screen to specifically identify loci critical for Ras1 signaling and further confirming a role for KSR in Ras1 signaling. In addition, we have identified 4 additional complementation groups. One of them corresponds to the kismet locus, which encodes a putative chromatin remodeling factor. The relevance of these loci with respect to the function of KSR and the: Ras1 pathway in general is discussed. C1 Univ Calif Berkeley, Howard Hughes Med Inst, Berkeley, CA 94720 USA. Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Mol Basis Carcinogenesis Lab, Frederick, MD 21702 USA. RP Rubin, GM (reprint author), Univ Calif Berkeley, Howard Hughes Med Inst, 545 Life Sci Addit 3200, Berkeley, CA 94720 USA. RI Tang, Amy/L-3226-2016; OI Tang, Amy/0000-0002-5772-2878; Rubin, Gerald/0000-0001-8762-8703 NR 45 TC 59 Z9 60 U1 0 U2 4 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 USA SN 0016-6731 J9 GENETICS JI Genetics PD NOV PY 2000 VL 156 IS 3 BP 1231 EP 1242 PG 12 WC Genetics & Heredity SC Genetics & Heredity GA 370VK UT WOS:000165143200025 PM 11063697 ER PT J AU Dunson, DB Tindall, KR AF Dunson, DB Tindall, KR TI Bayesian analysis of mutational spectra SO GENETICS LA English DT Article ID HISTORICAL CONTROLS; TRANSGENIC MICE; BINARY DATA; TESTS; MODELS; TREND; MUTANT; ASSAY; LACI; VARIABILITY AB Studies that examine both the frequency of gene mutation and the pattern or spectrum of mutational changes can be used to identify chemical mutagens and to explore the molecular mechanisms of mutagenesis. In this article, we propose a Bayesian hierarchical modeling approach for the analysis of mutational spectra. We assume that the total number of independent mutations and the numbers of mutations falling into different response categories, defined by location within a gene and/or type of alteration, follow binomial and multinomial sampling distributions, respectively. We use prior distributions to summarize past information about the overall mutation frequency and the probabilities corresponding to the different mutational categories. These priors call be chosen on the basis of data from previous studies using an approach that accounts for heterogeneity among studies. Inferences about the overall mutation frequency, the proportions of mutations in each response category, and the category-specific mutation frequencies can be based on posterior distributions, which incorporate past and current data on the mutant frequency and on DNA sequence alterations. Methods are described for comparing groups and for assessing dose-related trends. We illustrate our approach using data from the literature. C1 NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Res Triangle Pk, NC 27709 USA. RP Dunson, DB (reprint author), NIEHS, Biostat Branch, MD A3-03,POB 12233, Res Triangle Pk, NC 27709 USA. NR 31 TC 9 Z9 9 U1 0 U2 2 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 USA SN 0016-6731 J9 GENETICS JI Genetics PD NOV PY 2000 VL 156 IS 3 BP 1411 EP 1418 PG 8 WC Genetics & Heredity SC Genetics & Heredity GA 370VK UT WOS:000165143200040 PM 11063712 ER PT J AU Onyango, P Miller, W Lehoczky, J Leung, CT Birren, B Wheelan, S Dewar, K Feinberg, AP AF Onyango, P Miller, W Lehoczky, J Leung, CT Birren, B Wheelan, S Dewar, K Feinberg, AP TI Sequence and comparative analysis of the mouse 1-megabase region orthologous to the human 11p15 imprinted domain SO GENOME RESEARCH LA English DT Article ID BECKWITH-WIEDEMANN-SYNDROME; SNRPN GENE; CPG ISLAND; H19 GENE; METHYLATION; EXPRESSION; IGF2; GENOME; TUMOR; TRANSCRIPT AB A major barrier to conceptual advances in understanding the mechanisms and regulation of imprinting of a genomic region is our relatively poor understanding of the overall organization of genes and of the potentially important cis-acting regulatory sequences that lie in the nonexonic segments that make up 97% of the genome. Interspecies sequence comparison offers an effective approach to identify sequence from conserved functional elements. In this article we describe the successful use of this approach in comparing a similar to1-Mb imprinted genomic domain on mouse chromosome 7 to its orthologous region on human 11p15.5. Within the region, we identified 112 exons of known genes as well as a novel gene identified uniquely in the mouse region, termed Msuit, that was found to be imprinted. In addition to these coding elements, we identified 33 CpG islands and 49 orthologous nonexonic, nonisland sequences that met our criteria as being conserved, and making up 4.1% of the total sequence. These conserved noncoding sequence elements were generally clustered near imprinted genes and the majority were between Igf2 and H19 or within Kvlqt1. Finally, the location of CpG islands provided evidence that suggested a two-island rule for imprinted genes. This study provides the first global view of the architecture of an entire imprinted domain and provides candidate sequence elements for subsequent functional analyses. C1 Johns Hopkins Univ, Sch Med, Inst Med Genet, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. MIT, Ctr Genome Res, Whitehead Inst, Cambridge, MA 02141 USA. Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Oncol, Baltimore, MD 21205 USA. NIH, Ctr Biotechnol Informat, Bethesda, MD 20894 USA. RP Feinberg, AP (reprint author), Johns Hopkins Univ, Sch Med, Inst Med Genet, Baltimore, MD 21205 USA. NR 48 TC 105 Z9 108 U1 0 U2 1 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD NOV PY 2000 VL 10 IS 11 BP 1697 EP 1710 DI 10.1101/gr.161800 PG 16 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 374VH UT WOS:000165364700008 PM 11076855 ER PT J AU Lin, HC Pizer, ES Morin, PJ AF Lin, HC Pizer, ES Morin, PJ TI A frequent deletion polymorphism on chromosome 22q13 identified by representational difference analysis of ovarian cancer SO GENOMICS LA English DT Article ID CONVERTING-ENZYME GENE; DISEASE AB Sizable homozygous deletions (>100 bp) of genomic DNA in cancer cells are typically interpreted as an indication of the location of a tumor suppressor gene. In an effort to identify novel ovarian growth-suppressing genes, we performed representational difference analysis (RDA) of ovarian cancer cells. One of the RDA probes identified a 276-bp region of chromosome 22q deleted in 47% of the ovarian cancer cell lines examined. This small deletion was also found in the genomic DNA of 25% of colon cancer cell lines examined and, surprisingly, in 18% of the blood DNA samples from healthy controls. The deleted allele, which was named u22q, has a frequency of approximately 50% in the population and is in Hardy-Weinberg equilibrium with the intact allele. The deleted DNA sequence is flanked by direct repeats and likely originated through a slipped mispairing mechanism. The deletion did not encompass known transcripts or expressed sequence tags. It therefore appears likely that u22q represents a common polymorphism, often hemizygous in ovarian cancer because of a high rate of LOH of chromosome 22q. These findings provide an example of a sizable homozygous deletion that does not appear to be associated with disease. Such a finding provides a cautionary tale for positional cloning projects initiated exclusively on the basis of the identification of homozygous deletions. The possibility that the deletions in question may be constitutive should always be considered since it is probable that the genome contains a large number deletions/insertions of various sizes. (C) 2000 Academic Press. C1 NIA, Biol Chem Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Johns Hopkins Med Inst, Dept Pathol, Baltimore, MD 21287 USA. RP Morin, PJ (reprint author), NIA, Biol Chem Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 13 TC 9 Z9 9 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD NOV 1 PY 2000 VL 69 IS 3 BP 391 EP 394 DI 10.1006/geno.2000.6357 PG 4 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 371GB UT WOS:000165169500013 PM 11056057 ER PT J AU Owsley, C Burton-Danner, K Jackson, GR AF Owsley, C Burton-Danner, K Jackson, GR TI Aging and spatial localization during feature search SO GERONTOLOGY LA English DT Article DE aging; visual search; vision ID VISUAL-SEARCH; AGE-DIFFERENCES; FEATURE-INTEGRATION; GUIDED SEARCH; USEFUL FIELD; ATTENTION; MODEL; VIEW AB Background: Previous studies indicate that older adults, like young adults, can efficiently search for a briefly presented visual target defined by a single salient feature presented amidst background distracters. However, little is known about older adults' ability to identify the spatial location of targets during this aspect of preattentive processing. Objective: This study examined the extent to which older adults exhibit localization problems during feature search for a target with high conspicuity. Their performance was compared to that of younger adults. Methods: Twenty older adults (mean age 70 years, 8 men and 12 women) and 20 younger adults (mean age 25 years, 6 men and 14 women) with good central and peripheral vision were tested. Subjects were asked to indicate via a computerized touch-screen the location of a briefly presented (80 ms) target presented amidst distracting stimuli (set size 8, 16, or 32). Targets were presented at either 10 degrees, 20 degrees, or 30 degrees eccentricity. The dependent dent measures were percent correct localization and, for trials in which there were errors, the spatial magnitude of the error. Results: Compared to young adults, older adults committed more localization errors during feature search, a problem which was accentuated with increasing target eccentricity. In addition, older adults' mislocalizations deviated from the correct location by greater distances. Conclusions: Older adults have spatial localization problems in preattentive processing during feature search, which could be detrimental to the guidance and deployment of visual attention. Copyright (C) 2000 S. Karger AG, Basel. C1 Univ Alabama, Dept Ophthalmol, Sch Med, Birmingham, AL 35294 USA. NIH, Bethesda, MD 20892 USA. RP Owsley, C (reprint author), Univ Alabama, Dept Ophthalmol, Sch Med, 700 S 18th St,Suite 609, Birmingham, AL 35294 USA. RI Owsley, Cynthia/B-7986-2014 FU NIA NIH HHS [R01-AG04212, R03-AG12787] NR 38 TC 23 Z9 23 U1 2 U2 2 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 0304-324X J9 GERONTOLOGY JI Gerontology PD NOV-DEC PY 2000 VL 46 IS 6 BP 300 EP 305 DI 10.1159/000022181 PG 6 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 369BL UT WOS:000090151900002 PM 11044783 ER PT J AU Fabrega, SL Durand, P Codogno, P Bauvy, C Delomenie, C Henrissat, B Martin, BM McKinney, C Ginns, EI Mornon, JP Lehn, P AF Fabrega, SL Durand, P Codogno, P Bauvy, C Delomenie, C Henrissat, B Martin, BM McKinney, C Ginns, EI Mornon, JP Lehn, P TI Human glucocerebrosidase: heterologous expression of active site mutants in murine null cells SO GLYCOBIOLOGY LA English DT Article DE active site residues; catalytic machinery; Gaucher disease; glycoside hydrolases; human glucocerebrosidase; lysosomal enzymes; site-directed mutagenesis ID HUMAN BETA-GLUCOCEREBROSIDASE; HYDROPHOBIC CLUSTER-ANALYSIS; GLYCOSYL HYDROLASES; GAUCHERS-DISEASE; ACID/BASE CATALYST; IDENTIFICATION; SEQUENCE; GENE; FIBROBLASTS; GLUCOSIDASE AB Using bioinformatics methods, we have previously identified Glu235 and Glu340 as the putative acid/base catalyst and nucleophile? respectively, in the active site of human glucocerebrosidase. Thus, rye undertook site-directed mutagenesis studies to obtain experimental evidence supporting these predictions. Recombinant retroviruses were used to express wild-type and E235A and E340A mutant proteins in glucocerebrosidase-deficient murine cells. In contrast to wild-type enzyme, the mutants were found to be catalytically inactive. We also report the results of various studies (Western blotting, glycosylation analysis, subcellular fractionation, and confocal microscopy) indicating that the wild-type and mutant enzymes are identically processed and sorted to the lysosomes, Thus, enzymatic inactivity of the mutant proteins is not the result of incorrect folding/processing. These findings indicate that Glu235 plays a keg role in the catalytic machinery of human glucocerebrosidase and may indeed be the acidi base catalyst. As concerns Glu340, the results both support our computer-based predictions and confirm, at the biological level, previous identification of Glu340 as the nucleophile by use of active site labeling techniques. Finally, our findings may help to better understand the molecular basis of Gaucher disease, the human lysosomal disease resulting from deficiency in glucocerebrosidase. C1 Hop Robert Debre, INSERM U 458, F-75019 Paris, France. Univ Paris 06, CNRS UMR 7590, Lab Mineral Cristallog, Syst Mol & Biol Struct, F-75252 Paris 5, France. Univ Paris 07, CNRS UMR 7590, Lab Mineral Cristallog, Syst Mol & Biol Struct, F-75252 Paris 5, France. INSERM U 504, F-94807 Villejuif, France. Architecture & Fonct Macromol Biol, CNRS UPR 9039, F-13402 Marseille 20, France. NIMH, Clin Neurosci Branch, Bethesda, MD 20892 USA. RP Lehn, P (reprint author), Hop Robert Debre, INSERM U 458, 48 Bd Serurier, F-75019 Paris, France. RI Henrissat, Bernard/J-2475-2012; Codogno, Patrice/G-1384-2013 OI Codogno, Patrice/0000-0002-5492-3180 NR 43 TC 19 Z9 19 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD NOV PY 2000 VL 10 IS 11 BP 1217 EP 1224 DI 10.1093/glycob/10.11.1217 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 374VA UT WOS:000165364000011 PM 11087714 ER PT J AU Seeff, L AF Seeff, L TI Treatment of hepatitis C and prospects for future treatments SO HAEMOPHILIA LA English DT Article; Proceedings Paper CT XXIVth International Congress of the World-Federation-of-Hemophilia CY JUL 16-21, 2000 CL MONTREAL, CANADA SP World Federat Hemophilia C1 NIDDK, NIH, Bethesda, MD 20892 USA. RP Seeff, L (reprint author), NIDDK, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 1351-8216 J9 HAEMOPHILIA JI Haemophilia PD NOV PY 2000 VL 6 SU 2 BP 21 EP 21 PG 1 WC Hematology SC Hematology GA 382BG UT WOS:000165798100030 ER PT J AU Papoutsi, M Siemeister, G Weindel, K Tomarev, SI Kurz, H Schachtele, C Martiny-Baron, G Christ, B Marme, D Wilting, J AF Papoutsi, M Siemeister, G Weindel, K Tomarev, SI Kurz, H Schachtele, C Martiny-Baron, G Christ, B Marme, D Wilting, J TI Active interaction of human A375 melanoma cells with the lymphatics in vivo SO HISTOCHEMISTRY AND CELL BIOLOGY LA English DT Article DE lymphangiogenesis; tumor; chorioallantoic membrane; VEGF-C; Prox 1; lymphatic invasion; avian embryo ID ENDOTHELIAL GROWTH-FACTOR; CHICK CHORIOALLANTOIC MEMBRANE; VASCULAR-PERMEABILITY FACTOR; BLOOD-VESSEL INVASION; FACTOR-C EXPRESSION; VEGF-C; PROGNOSTIC-SIGNIFICANCE; TUMOR ANGIOGENESIS; TYROSINE KINASES; NODE METASTASIS AB We have used the avian chorioallantoic membrane (CAM) to study the interaction of tumor cells with the lymphatics in vivo. The vascular endothelial growth factor-C (VEGF-C) has been shown to be lymphangiogenic. We have therefore grown VEGF-C-expressing human A375 melanoma cells on the CAM. These tumors induced numerous lymphatics at the invasive front, and compressed or destroyed VEGF receptor (R)-3-positive lymphatics were observed within the sol id tumors. The lymphatics in the CAM and in the A375 melanomas could also be demonstrated with an antibody against Prox 1, a highly specific marker of lymphatic endothelial cells. Proliferation studies revealed a BrdU labeling index of 11.6% of the lymphatic endothelial cells in the tumors and at their margins. A great number of melanoma cells invaded the lymphatics. Such interactions were not observed with VEGF-C-negative Malme 3 M melanoma cells. Lymphangiogenesis was inhibited to some extent when A375 melanoma cells were transfected with cDNA encoding soluble VEGFR-3 (sflt4), and the BrdU labeling index of the lymphatics in these tumors was 3.9%. Invasion of lymphatics and growth of blood vascular capillaries were not inhibited by the transfection. Therefore, tumor-induced lymphangiogenesis seems to be dependent to some extent on VEGF-C/flt4 interactions, but invasion of lymphatics seems to be a distinct mechanism. C1 Univ Freiburg, Inst Anat, Lehrstuhl 2, D-79104 Freiburg, Germany. Univ Freiburg, Inst Mol Med & Nat Forsch, D-79106 Freiburg, Germany. NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Wilting, J (reprint author), Univ Freiburg, Inst Anat, Lehrstuhl 2, Albertstr 17, D-79104 Freiburg, Germany. EM wilting@uni-freiburg.de RI Kurz, Haymo/E-1587-2011 FU NICHD NIH HHS [N01-HD-6-2915] NR 68 TC 64 Z9 67 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0948-6143 J9 HISTOCHEM CELL BIOL JI Histochem. Cell Biol. PD NOV PY 2000 VL 114 IS 5 BP 373 EP 385 PG 13 WC Cell Biology; Microscopy SC Cell Biology; Microscopy GA 381FV UT WOS:000165752400004 PM 11151407 ER PT J AU Gladen, BC Zadorozhnaja, TD Chislovska, N Hryhorczuk, DO Kennicutt, MC Little, RE AF Gladen, BC Zadorozhnaja, TD Chislovska, N Hryhorczuk, DO Kennicutt, MC Little, RE TI Polycyclic aromatic hydrocarbons in placenta SO HUMAN & EXPERIMENTAL TOXICOLOGY LA English DT Article DE polycyclic aromatic hydrocarbons; placenta; birth weight ID UMBILICAL-CORD BLOOD; DNA-ADDUCTS; AIR-POLLUTION; URINARY 1-HYDROXYPYRENE; GENERAL-POPULATION; BREAST-MILK; 2 CITIES; EXPOSURE; BENZOPYRENE; POLAND AB Concentrations of chrysene, benz [a] anthracene, benzo [a]-pyrene, benzo[b]fluoranthene, indeno[1,2,3 -c,d]pyrene, dibenz[a,h] anthracene, and benzo[g,h,i] perylene were measured in placentas from 200 women from two cities in Ukraine, Kyiv and Dniprodzerzhinsk. The participants had no special exposures and were chosen from among subjects in an ongoing study of reproductive health. All seven of the polycyclic aromatic hydrocarbons (PAHs) were found in all placentas, with the sole exception of benzo [a] pyrene in one placenta. Chrysene was present at the highest concentrations, with median 1.38 ng/g dry weight. Dibenz [a,h] anthracene and benzo [g,h,i]perylene had the lowest concentrations; each had median 0.73 ng/g dry weight. Concentrations in Kyiv were slightly higher than those in Dniprodzerzhinsk, but the difference was significant only for dibenz[ a,h]anthracene. Dibenz[a,h] anthracene and benzo[g,h,i]perylene increased significantly with maternal body mass index, but other PAHs showed no such pattern. Placentas from deliveries in autumn or winter had slightly bur not significantly higher concentrations. Concentrations were not related to maternal age. There were too few smokers in the sample for meaningful evaluation. No associations were seen between any of the placental PAH concentrations and birth weight of the infant. C1 NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. Inst Pediat Obstet & Gynecol, Kiev, Ukraine. Univ Illinois, Sch Publ Hlth, Chicago, IL USA. Texas A&M Univ, Geochem & Environm Res Grp, College Stn, TX USA. RP Gladen, BC (reprint author), NIEHS, Biostat Branch, Mail Drop A3-03,POB 12233, Res Triangle Pk, NC 27709 USA. NR 41 TC 32 Z9 35 U1 2 U2 8 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0144-5952 J9 HUM EXP TOXICOL JI Hum. Exp. Toxicol. PD NOV PY 2000 VL 19 IS 11 BP 597 EP 603 DI 10.1191/096032700671433928 PG 7 WC Toxicology SC Toxicology GA 397FX UT WOS:000166683600001 PM 11211237 ER PT J AU Ben-Shaul, V Lomnitski, L Nyska, A Carbonatto, M Peano, S Zurovsky, Y Bergman, M Eldridge, SR Grossman, S AF Ben-Shaul, V Lomnitski, L Nyska, A Carbonatto, M Peano, S Zurovsky, Y Bergman, M Eldridge, SR Grossman, S TI Effect of natural antioxidants and apocynin on LPS-induced endotoxemia in rabbit SO HUMAN & EXPERIMENTAL TOXICOLOGY LA English DT Article DE inflammation; liver; oxidation stress; NADPH oxidase ID TUMOR-NECROSIS-FACTOR; LIPID-PEROXIDATION; SEPTIC SHOCK; BACTERIAL LIPOPOLYSACCHARIDE; ISCHEMIA-REPERFUSION; LABORATORY MODELS; ENDOTHELIAL-CELLS; FREE-RADICALS; NITRIC-OXIDE; LIVER AB The objective of this study was to compare the prophylactic effects of the natural antioxidant from spinach (NAO) and apocynin, on the hepatic oxidative stress and liver damage induced by lipopolysaccharide (LPS). Male New Zealand rabbits were challenged with LPS with or without 8 days of antioxidant pretreatment. Pretreatment with NAG, but not apocynin, significantly (p < 0.05) decreased the levels of hydroperoxides and malondialdehyde (MDA) in the liver cytosolic fraction and the activity of NADPH oxidase-generated superoxide in the microsomal fraction, compared to LPS alone. The activity of glutathione peroxidase (G-POX) was significantly (p < 0.05) increased in the LPS-treated group, whereas treatment with NAG, but not apocynin, significantly (p < 0.05) decreased G-POX activity. Pretreatment with the same antioxidants had no significant effects on superoxide dismutase (SOD) activity, whereas an increased level of catalase (CAT) was obtained in all LPS-treated groups. TUNEL immunohistochemical staining in the LPS- treated animals indicated that there was no increase in apoptosis outside of necrotic foci. However, apoptotic hepatocytes were observed within areas of focal necrosis in animals exposed to LPS alone or LPS plus apocynin. Hepatocyte cell proliferation was tested by the proliferating-cell nuclear antigen (PCNA) tool, which indicated a proliferative effect in the LPS group, whereas the effect disappeared in die antioxidant - treated groups. The prophylactic effect of NAO on liver pathology and the significant decreases in lipid peroxidation products and NADPH oxidase activity suggest the use of NAO as an efficient strategy for treatment of endotoxemia. C1 Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. NIEHS, Res Triangle Pk, NC 27709 USA. RBM, Inst Ric Biomed, LCG, I-10015 Turin, Italy. RP Grossman, S (reprint author), Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. NR 58 TC 15 Z9 17 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0144-5952 J9 HUM EXP TOXICOL JI Hum. Exp. Toxicol. PD NOV PY 2000 VL 19 IS 11 BP 604 EP 614 DI 10.1191/096032700666138364 PG 11 WC Toxicology SC Toxicology GA 397FX UT WOS:000166683600002 PM 11211238 ER PT J AU Vogel, JC AF Vogel, JC TI Nonviral skin gene therapy SO HUMAN GENE THERAPY LA English DT Article; Proceedings Paper CT Meeting on Cutaneous Gene Therapy: Problems and Prospects CY MAR 23-24, 2000 CL NIH, BETHESDA, MARYLAND SP Adv Tissue Sci, AGI Dermat, CIRD GALDERMA, Genzyme Corp, Ist Detmatopat Immacolata, Johnson & Johnson, Maxygen, Organogenesis Inc, Procter & Gamble HO NIH ID IMMUNOSTIMULATORY DNA-SEQUENCES; DENDRITIC CELLS; IN-VIVO; BACTERIAL-DNA; IMMUNE-RESPONSE; CPG MOTIFS; NAKED DNA; LIPOSOME COMPLEXES; LEISHMANIA-MAJOR; MURINE MELANOMA AB Nonviral skin gene therapy is an effective method to directly deliver and transiently express genes in the skin. Several different nonviral delivery methods have been successfully used and are analyzed here for their efficiency and efficacy in achieving specific therapeutic applications. For one important and frequently used application of nonviral skin gene therapy, genetic immunization, the types of resulting immune responses (Th1 versus Th2) will depend on which delivery method is used. In addition, we discuss the contributions of DNA as an immunostimulatory adjuvant in genetic immunization and how activation of skin dendritic cells and induction of IL-12 expression are mechanistically important in this process. Nonviral skin gene therapy has also been successfully used to enhance tumor regression in animal models, frequently by inducing a specific immune response against the tumor. In the future, nonviral skin gene therapy may be successfully used for the treatment of additional skin diseases if genes can be selectively delivered and expressed in specific skin cells, and if increased level and duration of gene expression can be achieved. C1 NCI, Dermatol Branch, NIH, Bethesda, MD 20892 USA. RP Vogel, JC (reprint author), NCI, Dermatol Branch, NIH, 10 Ctr Dr,Bldg 10, Bethesda, MD 20892 USA. NR 56 TC 55 Z9 56 U1 0 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1043-0342 J9 HUM GENE THER JI Hum. Gene Ther. PD NOV 1 PY 2000 VL 11 IS 16 BP 2253 EP 2259 DI 10.1089/104303400750035780 PG 7 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 374DG UT WOS:000165328700007 PM 11084683 ER PT J AU Parker, LL Do, MT Westwood, JA Wunderlich, JR Dudley, ME Rosenberg, SA Hwu, P AF Parker, LL Do, MT Westwood, JA Wunderlich, JR Dudley, ME Rosenberg, SA Hwu, P TI Expansion and characterization of T cells transduced with a chimeric receptor against ovarian cancer SO HUMAN GENE THERAPY LA English DT Article ID TUMOR-INFILTRATING LYMPHOCYTES; FOLATE-BINDING PROTEIN; MONOCLONAL-ANTIBODY; ANTITUMOR-ACTIVITY; IMMUNE-RESPONSES; GENE-TRANSFER; INTERLEUKIN-2; MELANOMA; THERAPY; GAMMA AB Adoptive immunotherapy with genetically modified T lymphocytes is being utilized in clinical trials for the treatment of a broad range of diseases including cancer and HIV infection. To improve on these treatments, and to better understand their mechanisms of action, it is necessary to develop techniques to generate large numbers of cells and characterize the functional heterogeneity of the cells produced. In this study, patient peripheral blood lymphocytes were transduced with a chimeric antigen receptor (MOv-gamma) derived from a mouse monoclonal antibody against folate-binding protein, which is overexpressed on many ovarian cancers. Thus, irrespective of their original specificity, normal human T lymphocytes were redirected to react against ovarian cancer cells. Lymphocytes from five patients were transduced and grown to large numbers, with a median expansion of more than 7000-fold. When proliferation was inadequate, the cells were expanded by stimulation utilizing anti-CD3, IL-2, and irradiated allogeneic PBMCs. The cells maintained their functional ability to recognize ovarian cancer over several months. Cloning of transduced cells was undertaken to determine the level of gene expression and function of individual cells making up the bulk population. Transduced CD4(+) and CD8(+) cell clones were isolated from the bulk and demonstrated antitumor activity. These clones had a diverse repertoire with respect to secretion of cytokines, and individual clones maintained their cytokine profile on subsequent expansion. These studies establish the feasibility of consistently generating large numbers of gene-modified tumor-reactive lymphocytes, with a stable and diverse cytokine repertoire, that could be utilized for patient treatment. C1 NCI, Tumor Immunol Sect, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Hwu, P (reprint author), NCI, Tumor Immunol Sect, Surg Branch, NIH, Bldg 10,Room 2B42,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 39 TC 36 Z9 37 U1 0 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1043-0342 J9 HUM GENE THER JI Hum. Gene Ther. PD NOV PY 2000 VL 11 IS 17 BP 2377 EP 2387 DI 10.1089/104303400750038480 PG 11 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 376AV UT WOS:000165436400006 PM 11096442 ER PT J AU Wolford, JK Blunt, D Ballecer, C Prochazka, M AF Wolford, JK Blunt, D Ballecer, C Prochazka, M TI High-throughput SNP detection by using DNA pooling and denaturing high performance liquid chromatography (DHPLC) SO HUMAN GENETICS LA English DT Article ID MUTATION DETECTION; PIMA-INDIANS; POLYMORPHISMS AB One of the critical steps in the positional cloning of a complex disease gene involves association analysis between a phenotype and a set of densely spaced diallelic markers, typically single nucleotide repeats (SNPs), covering the region of interest. However, the effort and cost of detecting sufficient numbers of SNPs across relatively large physical distances represents a significant rate-limiting step. We have explored DNA pooling, in conjunction with denaturing high performance liquid chromatography (DHPLC), as a possible strategy for augmenting the efficiency, economy, and throughput of SNP detection. DHPLC is traditionally used to detect variants in polymerase chain reaction products containing both allelic forms of a polymorphism (e.g., heterozygotes or a 1:1 mix of both alleles) via heteroduplex separation and thereby requires separate analyses of multiple individual test samples. We have adapted this technology to identify variants in pooled DNA. To evaluate the utility and sensitivity of this approach, we constructed DNA pools comprised of 20 previously genotyped individuals with a frequency representation of 0%-50% for the variant allele. Mutation detection was performed by using temperature-modulated heteroduplex formation/DHPLC and dye-terminator sequencing. Using DHPLC, we could consistently detect SNPs at lower than 5% frequency, corresponding to the detection of one variant allele in a pool of 20 alleles. In contrast, fluorescent sequencing detected variants in the same pools only if the frequency of the less common allele was at least 10%. We conclude that DNA pooling of samples for DHPLC analysis is an effective way to increase throughput efficiency of SNP detection. C1 NIDDKD, Clin Diabet & Nutr Sect, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ 85016 USA. RP Wolford, JK (reprint author), NIDDKD, Clin Diabet & Nutr Sect, Phoenix Epidemiol & Clin Res Branch, NIH, 4212 N 16Th St, Phoenix, AZ 85016 USA. NR 20 TC 88 Z9 106 U1 1 U2 5 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD NOV PY 2000 VL 107 IS 5 BP 483 EP 487 DI 10.1007/s004390000396 PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 381FP UT WOS:000165751900008 PM 11140946 ER PT J AU Winterer, G Smolka, M Samochowiec, J Mulert, C Ziller, M Mahlberg, R Wuebben, Y Gallinat, J Rommelspacher, H Herrmann, WM Sander, T AF Winterer, G Smolka, M Samochowiec, J Mulert, C Ziller, M Mahlberg, R Wuebben, Y Gallinat, J Rommelspacher, H Herrmann, WM Sander, T TI Association analysis of GABA(A) beta 2 and gamma 2 gene polymorphisms with event-related prefrontal activity in man SO HUMAN GENETICS LA English DT Article ID ELECTROMAGNETIC TOMOGRAPHY; ALCOHOL DEPENDENCE; EEG; BRAIN; HIPPOCAMPUS; POTENTIALS; VARIANTS; SUBUNIT; CLUSTER AB Gamma-aminobutyric acid (GABA)(A)-receptors play a crucial role in the generation of electroencephalogram (EEC) oscillations and evoked potentials (ERPs). The present association study was designed to test whether EEG and ERPs are modulated by genetic variations of the human GABA(A) beta2 (GABRB2) and gamma2 (GABRG2) genes on chromosome 5q33. The genotypes of two nucleotide substitution polymorphisms of the GABRB2 and GABRG2 genes were assessed in 95 psychiatrically healthy subjects of German descent. Neurophysiological phenotyping was performed with four factorized EEG/ERP parameters: EEG activation, anterior and posterior EEG synchronization, and event-related activity (N100/P200-complex). No genotypic association was found for the GABRB2 nucleotide exchange polymorphism with any electrophysiological parameter. A significant association was found between the genotype of the intronic GABRG2 G-->A nucleotide exchange and the event-related N100/P200 (ANOVA: F=3.81; df=2; P=0.026). A comparison of homozygous subjects carrying either the G/G or A/A genotype of the GABRG2 polymorphism consistently revealed an even stronger difference in the effect-size (ANOVA: F=11.13; df=1; P=0.002). Post hoc analysis of this association with current density analysis in three-dimensional neuroanatomic Talairach space-time showed a reduction in the event-related signal power after 120 ms in the right dorsolateral prefrontal cortex. Taking into account the risk of false-positive association findings attributable to multiple testing, our results encourage further replication studies to examine the phenotype-genotype relationship of GABRG2 gene variants and event-related prefrontal activity. C1 Free Univ Berlin, Hosp Benjamin Franklin, Dept Psychiat, Berlin, Germany. Humboldt Univ, Univ Hosp, Charite, Dept Neurol, Berlin, Germany. NIMH, Clin Brain Disorder Branch, NIH, Bethesda, MD 20892 USA. RP Winterer, G (reprint author), Free Univ Berlin, Hosp Benjamin Franklin, Dept Psychiat, Berlin, Germany. RI Smolka, Michael/B-4865-2011; Mulert, Christoph/F-2576-2012; Samochowiec, Jerzy/G-8175-2014 OI Smolka, Michael/0000-0001-5398-5569; Samochowiec, Jerzy/0000-0003-1438-583X NR 32 TC 18 Z9 18 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD NOV PY 2000 VL 107 IS 5 BP 513 EP 518 DI 10.1007/s004390000401 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 381FP UT WOS:000165751900013 PM 11140951 ER PT J AU Lin, BC Pan, CJ Chou, JY AF Lin, BC Pan, CJ Chou, JY TI Human variant glucose-6-phosphate transporter is active in microsomal transport SO HUMAN GENETICS LA English DT Article ID DISEASE TYPE 1B; TRANSMEMBRANE TOPOLOGY AB Glycogen storage disease type 1b (GSD-1b) is caused by deficiencies in the glucose-6-phosphate transporter (G6PT), which works together with glucose-6-phosphatase to maintain glucose homeostasis. In humans, there are two alternatively spliced transcripts, G6PT and variant G6PT (vG6PT), differing by the inclusion of a 66-bp exon-7 sequence in vG6PT. We have previously shown that the G6PT protein functions as a microsomal glucose-6-phosphate (G6P) transporter, which is anchored to the endoplasmic reticulum by ten transmembrane helices. Here, we demonstrate that vG6PT is also active in microsomal G6P transport. The additional 22 amino acids in vG6PT is predicted to constitute a part of the luminal loop 4. Our data indicate that this loop plays no vital role in microsomal: G6P transport. Further, we show that G6PT mRNA is expressed in all organs and tissues examined, but that the vG6PT transcript is expressed exclusively in the brain, heart, and skeletal muscle. These results raise the possibility that mutations in exon-7 of the G6PT gene, which would not perturb glucose homeostasis, might have other deleterious effects. C1 NICHHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Chou, JY (reprint author), NICHHD, Heritable Disorders Branch, NIH, Bldg 10,Room 9S241, Bethesda, MD 20892 USA. RI Lin, Baochuan/A-8390-2009 OI Lin, Baochuan/0000-0002-9484-0785 NR 8 TC 19 Z9 19 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD NOV PY 2000 VL 107 IS 5 BP 526 EP 529 DI 10.1007/s004390000404 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 381FP UT WOS:000165751900015 PM 11140953 ER PT J AU Di Muzio, M Spoletini, L Strizzi, L Vianale, G Fontana, V Orengo, MA Tassi, G Casalini, A Mutti, L Procopio, A AF Di Muzio, M Spoletini, L Strizzi, L Vianale, G Fontana, V Orengo, MA Tassi, G Casalini, A Mutti, L Procopio, A TI Prognostic significance of presence and reduplication of basal lamina in malignant pleural mesothelioma SO HUMAN PATHOLOGY LA English DT Article DE mesothelioma; basal lamina; prognosis ID ADENOID CYSTIC CARCINOMA; ULTRASTRUCTURAL DISTINCTION; CONSECUTIVE PATIENTS; CANCER; DIFFERENTIATION; SURVIVAL AB The prognosis of patients with malignant pleural mesothelioma (MM) is dependent more on tumor extension and differentiation than on therapeutic effects. Reduplication of the basal lamina (RBL) is an ultrastructural feature of some benign and malignant tumors that has been inversely correlated with aggressiveness and was recently described in MM. To investigate whether RBL is important for predicting the survival of patients with MM, transmission electron microscopy was used to identify the presence of basal lamina or RBL in biopsy specimens obtained by thoracoscopy from 35 patients. Cox's regression analysis was used to study the relation of these ultrastructural features to survival. Better outcomes were found for patients whose tumors expressed either basal lamina (HR 0.48; 95% CI, 0.09-2.47) or RBL (HR 0.38; 95% CI 0.12-1.22) compared with the reference category, where basal lamina or RBL was not found. The expression of basal lamina and RBL is an important novel prognostic factors in MM. HUM PATHOL 31:141-1345. Copyright (C) 2000 by W.B. Saunders Company. C1 Univ G DAnnunzio, Dipartimento Oncol & Neurosci, Sez Patol Clin, I-66013 Chieti, Italy. Natl Canc Inst, Dept Environm Epidemiol & Biostat, Genoa, Italy. Natl Canc Inst, Liguria Canc Registry Sect, Genoa, Italy. Valle Camon Hosp, Div Resp Dis, Esine, Italy. Rasori Hosp, Bronchol & Thoracoscopy Serv, Parma, Italy. S Maugeri Fdn, Inst Hlth & Res, Pavia, Italy. RP Procopio, A (reprint author), Univ G DAnnunzio, Dipartimento Oncol & Neurosci, Sez Patol Clin, Edificio C,3 Piano,Via Dei Vestini 31, I-66013 Chieti, Italy. NR 33 TC 9 Z9 9 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0046-8177 J9 HUM PATHOL JI Hum. Pathol. PD NOV PY 2000 VL 31 IS 11 BP 1341 EP 1345 PG 5 WC Pathology SC Pathology GA 376JJ UT WOS:000165454200002 PM 11112207 ER PT J AU Mueller, SO Tahara, H Barrett, JC Korach, KS AF Mueller, SO Tahara, H Barrett, JC Korach, KS TI Immortalization of mammary cells from estrogen receptor alpha knock-out and wild-type mice SO IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL LA English DT Letter ID SMOOTH-MUSCLE DIFFERENTIATION; KERATIN GENE-EXPRESSION; TELOMERASE ACTIVITY; PRIMARY CULTURE; BREAST-CANCER; IN-VITRO; MOUSE; GROWTH; DISRUPTION; PROLIFERATION C1 NIEHS, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Korach, KS (reprint author), NIEHS, Reprod & Dev Toxicol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. EM korach@niehs.nih.gov NR 33 TC 1 Z9 1 U1 0 U2 0 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 1071-2690 EI 1543-706X J9 IN VITRO CELL DEV-AN JI In Vitro Cell. Dev. Biol.-Anim. PD NOV-DEC PY 2000 VL 36 IS 10 BP 620 EP 624 DI 10.1290/1071-2690(2000)036<0620:IOMCFE>2.0.CO;2 PG 5 WC Cell Biology; Developmental Biology SC Cell Biology; Developmental Biology GA 404GN UT WOS:000167089200002 PM 11229591 ER PT J AU Ruhl, S Cisar, JO Sandberg, AL AF Ruhl, S Cisar, JO Sandberg, AL TI Identification of polymorphonuclear leukocyte and HL-60 cell receptors for adhesins of Streptococcus gordonii and Actinomyces naeslundii SO INFECTION AND IMMUNITY LA English DT Article ID INFLUENZA-A VIRUS; PROTEIN-TYROSINE-PHOSPHATASE; TYPE-2 FIMBRIAL LECTIN; ACID-BINDING LECTINS; COMMON ANTIGEN CD45; HUMAN-NEUTROPHILS; SIALOPHORIN CD43; SURFACE SIALOGLYCOPROTEIN; SALIVARY GLYCOPROTEINS; POLYACRYLAMIDE GELS AB Interactions of oral streptococci and actinomyces with polymorphonuclear leukocytes (PMNs), mediated by sialic acid- and Gal/GalNAc-reactive adhesins, respectively, result in activation of the PMNs and thereby may contribute to the initiation of oral inflammation. Sialidase treatment of PMNs or HL-60 cells abolished adhesion of Streptococcus gordonii but was required for adhesion of Actinomyces naeslundii, The same effects of sialidase were noted for adhesion of these bacteria to a major 150-kDa surface glycoprotein of either PMNs or undifferentiated HL-60 cells and to a 130-kDa surface glycoprotein of differentiated HL-60 cells. These glycoproteins were both identified as leukosialin (CD43) by immunoprecipitation with a specific monoclonal antibody (MAb), Adhesion of streptococci and actinomyces to a 200-kDa minor PMN surface glycoprotein was also detected by bacterial overlay of untreated and sialidase-treated nitrocellulose transfers, respectively. This glycoprotein was identified as leukocyte common antigen (CD45) by immunoprecipitation with a specific MAb. CD43 and CD45 both possess extracellular mucinlike domains in addition to intracellular domains that are implicated in signal transduction, Consequently, the interactions of streptococci and actinomyces with the mucinlike domains of these mammalian cell surface glycoproteins result not only in adhesion but, in addition, may represent the initial step in PMN activation by these bacteria. C1 Natl Inst Dent & Craniofacial Res, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. RP Sandberg, AL (reprint author), Bldg 45,Room 4AN-24A, Bethesda, MD 20892 USA. NR 57 TC 36 Z9 36 U1 1 U2 4 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD NOV PY 2000 VL 68 IS 11 BP 6346 EP 6354 DI 10.1128/IAI.68.11.6346-6354.2000 PG 9 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 366LN UT WOS:000090007000035 PM 11035744 ER PT J AU Langer, RC Hayward, RE Tsuboi, T Tachibana, M Torii, M Vinetz, JM AF Langer, RC Hayward, RE Tsuboi, T Tachibana, M Torii, M Vinetz, JM TI Micronemal transport of Plasmodium ookinete chitinases to the electron-dense area of the apical complex for extracellular secretion SO INFECTION AND IMMUNITY LA English DT Article ID TRANSMISSION-BLOCKING ANTIBODIES; SURFACE PROTEIN; PARASITE CHITINASE; MALARIAL PARASITE; MOSQUITO; BERGHEI; GENE; GALLINACEUM; FALCIPARUM; ANTIGENS AB Plasmodium ookinetes secrete chitinases to penetrate the acellular, chitin-containing peritrophic matrix of the mosquito midgut en route to invasion of the epithelium. Chitinases are potentially targets that can be used to block malaria transmission. We demonstrate here that chitinases of Plasmodium falciparum and P. gallinaceum are concentrated at the apical end of ookinetes. The chitinase PgCHT1 of P. gallinaceum is present within ookinete micronemes and subsequently becomes localized in the electron-dense area of the apical complex. These observations suggest a pathway by which ookinetes secrete proteins extracellularly. C1 Univ Texas, Med Branch, WHO Collaborating Ctr Trop Dis, Dept Pathol, Galveston, TX 77555 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. Ehime Univ, Sch Med, Dept Mol Parasitol, Shigenobu, Ehime 7910295, Japan. RP Torii, M (reprint author), Univ Texas, Med Branch, WHO Collaborating Ctr Trop Dis, Dept Pathol, Keiller 2-138,301 Univ Blvd, Galveston, TX 77555 USA. OI Vinetz, Joseph/0000-0001-8344-2004 FU NIAID NIH HHS [T32-AI07536, T32 AI007536, R01-AI 45999, R01 AI045999] NR 26 TC 21 Z9 21 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD NOV PY 2000 VL 68 IS 11 BP 6461 EP 6465 DI 10.1128/IAI.68.11.6461-6465.2000 PG 5 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 366LN UT WOS:000090007000051 PM 11035760 ER PT J AU Chronwall, BM Sands, SA Cummings, KC Schwartz, JP AF Chronwall, BM Sands, SA Cummings, KC Schwartz, JP TI Glial somatostatin-14 expression in the rat pituitary intermediate lobe: a possible neurotrophic function during development? SO INTERNATIONAL JOURNAL OF DEVELOPMENTAL NEUROSCIENCE LA English DT Article DE glia-derived neurotrophic factor; axonal guidance; immunohistochemistry; RT-PCR ID TRANSIENT EXPRESSION; GFAP EXPRESSION; MESSENGER-RNA; INSITU HYBRIDIZATION; NEURITE OUTGROWTH; GENE-EXPRESSION; NEONATAL RAT; HYPOTHALAMUS; ONTOGENY; SYSTEM AB Somatostatin-14 was first detected on gestational day 17 in radially-oriented, bipolar cells spanning the width of the intermediate lobe of the rat pituitary. Cells were prominent, and constituted approximately 50% of the lobe area. The presence of vimentin, the cellular shape, and the localization identified these cells as glia. At postnatal day 6, somatostatin-14 and vimentin staining appeared in stellate-shaped cells. This is in agreement with the change from bipolar to stellate shape these glia undergo after the onset of innervation ([13] Gary et al. Int. J. Devl. Neurosci. 13, 555-565, 1995). Glia were more abundant, relative to melanotropes, throughout embryonic and early postnatal development compared to adulthood. Reverse transcription-polymerase chain reaction data showed a high level of prosomatostatin mRNA in the intermediate lobe: compared to the anterior and neural lobes fr om postnatal day 2 animals, and a significant drop in intermediate lobe content in the adult. The correlation between the number of glia and high expression of somatostatin in neonatal relative to adult tissue, together with the close apposition of incoming axons to the abundant, radially oriented glia during innervation of the lobe, support a neurotrophic function of glia-derived somatostatin. Published by Elsevier Science Ltd on behalf of ISDN. C1 Univ Missouri, Sch Biol Sci, Kansas City, MO 64110 USA. NINDS, Neurotroph Factors Sect, Bethesda, MD 20892 USA. RP Chronwall, BM (reprint author), Univ Missouri, Sch Biol Sci, 5100 Rockhill Rd, Kansas City, MO 64110 USA. EM chronwall@umkc.edu FU NINDS NIH HHS [NS 28019] NR 38 TC 7 Z9 7 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0736-5748 J9 INT J DEV NEUROSCI JI Int. J. Dev. Neurosci. PD NOV PY 2000 VL 18 IS 7 BP 685 EP 692 DI 10.1016/S0736-5748(00)00035-6 PG 8 WC Developmental Biology; Neurosciences SC Developmental Biology; Neurosciences & Neurology GA 360KE UT WOS:000089665900009 PM 10978847 ER PT J AU Hori, M Takechi, K Arai, Y Yomo, H Itabashi, M Shimazaki, J Inagawa, S Hori, M AF Hori, M Takechi, K Arai, Y Yomo, H Itabashi, M Shimazaki, J Inagawa, S Hori, M TI Comparison of macroscopic appearance and estrogen receptor-alpha regulators after gene alteration in human endometrial cancer SO INTERNATIONAL JOURNAL OF GYNECOLOGICAL CANCER LA English DT Article DE endometrial cancer; MDM2; p53; ras oncogene ID P53 PROTEIN; PROGNOSTIC INDICATOR; CARCINOMA; MDM2; EXPRESSION; MUTATIONS; OVEREXPRESSION; TRANSCRIPTS; ACTIVATION; SURVIVAL AB The existence of two types of endometrial cancer (hyperplasia-associated type [type I] and atrophy-associated type [type III) is well established. To test if different molecular genetic pathways are involved in the pathogenesis of type I and II disease, we examined pathologic features and the genetic alterations of K-Ras, MDM2 and p53 (which are considered to be involved in regulation of the estrogen receptor-alpha) in human endometrial tissue samples using several modified PCR methods. We found a significant difference in histologic grade (P < 0.001), degree of invasion (P < 0.001), stage grouping (P < 0.001) and estrogen receptor status (P < 0.01) between type I and II cases. There was a tendency for cases with K-Ras point mutations to be of type I, and for cases with the p53 point mutation to be of type II; however, we found that these mutations or alternative splicing of MDM2 was rarely involved and there was no significant difference in frequency of these alterations between types I and II. There must therefore be another structural or functional difference of Ras, MDM2 or p53 between type I and type II cancer. These unknown factors may be responsible for the difference between estrogen-dependent and estrogen-independent growth in human endometrial cancer. C1 Ibaraki Prefectural Cent Hosp & Canc Ctr, Dept Pathol, Ibaraki, Osaka 3091703, Japan. NIAID, Immunopathol Lab, NIH, Bethesda, MD 20892 USA. RP Hori, M (reprint author), Ibaraki Prefectural Cent Hosp & Canc Ctr, Dept Pathol, 6528 Koibuchi, Ibaraki, Osaka 3091703, Japan. EM ms-hori@chubyoin.pref.ibaraki.jp NR 30 TC 4 Z9 4 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1048-891X J9 INT J GYNECOL CANCER JI Int. J. Gynecol. Cancer PD NOV-DEC PY 2000 VL 10 IS 6 BP 469 EP 476 DI 10.1046/j.1525-1438.2000.00076.x PG 8 WC Oncology; Obstetrics & Gynecology SC Oncology; Obstetrics & Gynecology GA 394MK UT WOS:000166527700007 ER PT J AU Blegen, H Einhorn, N Sjovall, K Roschke, A Ghadimi, BM McShane, LM Nilsson, B Shah, K Ried, T Auer, G AF Blegen, H Einhorn, N Sjovall, K Roschke, A Ghadimi, BM McShane, LM Nilsson, B Shah, K Ried, T Auer, G TI Prognostic significance of cell cycle proteins and genomic instability in borderline, early and advanced stage ovarian carcinomas SO INTERNATIONAL JOURNAL OF GYNECOLOGICAL CANCER LA English DT Article DE carcinoma; cell cycle; genomic instability; ovary; prognosis ID BREAST-CANCER; CHROMOSOME-ABERRATIONS; P53 ACCUMULATION; TUMORS; SURVIVAL; EXPRESSION; P27(KIP1); P27 AB Disturbed cell cycle-regulating checkpoints and impairment of genomic stability are key events during the genesis and progression of malignant tumors. We analyzed 80 epithelial ovarian tumors of benign (n = 10) and borderline type (n = 18) in addition to carcinomas of early (n = 26) and advanced (n = 26) stages for the expression of Ki67, cyclin A and cyclin E, p21(WAF-1), p27(KIP-1) and p53 and correlated the results with the clinical course. Genomic instability was assessed by DNA ploidy measurements and, in 35 cases, by comparative genomic hybridization. Overexpression of cyclin A and cyclin E was observed in the majority of invasive carcinomas, only rarely in borderline tumors and in none of the benign tumors. Similarly, high expression of p53 together with undetectable p21 or loss of chromosome arm 17p were frequent events only in adenocarcinomas. Both borderline tumors and adenocarcinomas revealed a high number of chromosomal gains and losses. However, regional chromosomal amplifications were found to occur 13 times more frequently in the adenocarcinomas than in the borderline tumors. The expression pattern of low p27 together with high Ki67 was found to be an independent predictor of poor outcome in invasive carcinomas. The results provide a link between disturbed cell cycle regulatory proteins, chromosomal aberrations and survival in ovarian carcinomas. C1 Karolinska Hosp, Dept Pathol & Oncol, SE-17176 Stockholm, Sweden. Karolinska Hosp, Dept Gynecol, SE-17176 Stockholm, Sweden. Karolinska Hosp, Dept Gynecol Oncol, SE-17176 Stockholm, Sweden. NCI, Dept Genet, Div Clin Sci, Bethesda, MD 20892 USA. NCI, Biometr Res Branch, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Baltimore, MD USA. RP Auer, G (reprint author), Karolinska Hosp, Dept Pathol & Oncol, CCK R8-04, SE-17176 Stockholm, Sweden. NR 30 TC 36 Z9 36 U1 0 U2 4 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 1048-891X J9 INT J GYNECOL CANCER JI Int. J. Gynecol. Cancer PD NOV-DEC PY 2000 VL 10 IS 6 BP 477 EP 487 DI 10.1046/j.1525-1438.2000.00077.x PG 11 WC Oncology; Obstetrics & Gynecology SC Oncology; Obstetrics & Gynecology GA 394MK UT WOS:000166527700008 ER PT J AU Reitman, ML Gavrilova, O AF Reitman, ML Gavrilova, O TI A-ZIP/F-1 mice lacking white fat: a model for understanding lipoatrophic diabetes SO INTERNATIONAL JOURNAL OF OBESITY LA English DT Article; Proceedings Paper CT Symposium on Recent Advances in Adipose Tissue Biology and Their Implications for Obesity CY NOV 19-20, 1999 CL QUEBEC CITY, CANADA SP Hoffmann La Roche Canada DE lipoatrophy/lipodystrophy; diabetes; insulin resistance; adipose tissue; fasting; torpor; leptin ID CONGENITAL GENERALIZED LIPODYSTROPHY; ADIPOSE-TISSUE; INSULIN-RESISTANCE; LEPTIN; MELLITUS; GENE AB The A-ZIP/F-1 mouse is lacking virtually all white adipose tissue. Like humans with extensive deficiencies of adipose tissue, the A-ZIP/F-1 mice develop a severe form of insulin resistant diabetes. We have studied the physiology of the A-ZIP/F-1 mice. Their adaptation to fasting is notable for its rapidity and the use of torpor, a hibernation-like state, to minimize energy needs. Transplantation of adipose tissue reversed the metabolic manifestations in the mice, demonstrating that the lack of adipose tissue is the cause of the insulin resistance. Leptin replacement is not very effective in reversing the diabetes of the A-ZIP/F-1 mice, which contrasts with its efficacy in the aP2-SREBP-1c mouse. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Reitman, ML (reprint author), NIDDKD, Diabet Branch, NIH, Bldg 10,Room 8N-250, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013 OI Reitman, Marc/0000-0002-0426-9475 NR 19 TC 27 Z9 28 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0307-0565 J9 INT J OBESITY JI Int. J. Obes. PD NOV PY 2000 VL 24 SU 4 BP S11 EP S14 PG 4 WC Endocrinology & Metabolism; Nutrition & Dietetics SC Endocrinology & Metabolism; Nutrition & Dietetics GA 374CX UT WOS:000165327800005 PM 11126232 ER PT J AU Junker, K Moravek, P Podhola, M Weirich, G Hindermann, W Janitzky, V Schubert, J AF Junker, K Moravek, P Podhola, M Weirich, G Hindermann, W Janitzky, V Schubert, J TI Genetic alterations in metastatic renal cell carcinoma detected by comparative genomic hybridization: Correlation with clinical and histological data SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE kidney cancer; metastasis; genetics; comparative genomic hybridization ID ALLELIC LOSS; TUMORS; CLASSIFICATION; CANCERS; STAGE; GRADE; DNA; 14Q AB In order to optimize the management of patients with renal cell carcinoma (RCC) it is important to define the genetic risk for metastatic disease. In this study we performed comparative genomic hybridization (CGH) on metastatic tumors aiming at the identification of genetic alterations associated with metastatic disease. We analyzed 46 renal tumors along with their metastases, and 15 non-metastatic renal tumors. Tumors were classified pathologically according to the Heidelberg classification of RCC, and staged according to the TNM-system. Standard CGH was performed using microdissected archival tissues and DOP-PCR. The average numbers of chromosomal aberrations per tumor were 3.0, 2.1 and 3.9 in patients without metastasis, in patients who developed metastases after a two-year latency period (late onset of metastatic disease) and in patients who developed metastases within two years after therapy of the primary tumor (early onset of metastatic disease). CGH revealed chromosomal aberrations in 91% of primary metastatic tumors. Deletions or losses of chromosomes 9 (26% vs 6%), 10 (21% vs 6%) and 18 (23% vs 0) and 17 (28% vs 7%) occurred more often in metastatic tumors than in nonmetastatic tumors. Furthermore, these aberrations were more common in patients with early metastases. CGH analysis of 40 pairs of primary RCCs and their corresponding metastasis revealed similar aberrations in 70% of cases. In 30%, however, metastases showed additional chromosomal aberrations not detected in the corresponding primary tumors. In conclusion, we identified genetic alterations associated with metastatic disease in RCC which could be useful for predicting prognosis. Genetic changes leading to metastases occurred early in tumorigenesis of metastatic tumors. C1 Dept Urol, D-07743 Jena, Germany. Charles Univ, Inst Pathol, Hradec Kralove 50005 11, Czech Republic. Charles Univ, Dept Urol, Hradec Kralove 50005 11, Czech Republic. NCI, LIB, FCRDC, Frederick, MD 21702 USA. Inst Pathol, D-07743 Jena, Germany. RP Junker, K (reprint author), Dept Urol, Lessingstr 1, D-07743 Jena, Germany. NR 19 TC 24 Z9 24 U1 0 U2 1 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD NOV PY 2000 VL 17 IS 5 BP 903 EP 908 PG 6 WC Oncology SC Oncology GA 363FW UT WOS:000089825300005 PM 11029490 ER PT J AU Vodovotz, Y Waksman, R Cook, JA Kim, WH Chan, R Seabron, R Collins, SD Pierre, A Bramwell, O Wink, D Mitchell, JB Leon, MB AF Vodovotz, Y Waksman, R Cook, JA Kim, WH Chan, R Seabron, R Collins, SD Pierre, A Bramwell, O Wink, D Mitchell, JB Leon, MB TI S-nitrosoglutathione reduces nonocclusive thrombosis rate following balloon overstretch injury and intracoronary irradiation of porcine coronary arteries SO INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS LA English DT Article DE restenosis; radiation; swine; nitric oxide; thrombosis ID INHIBITS NEOINTIMA FORMATION; NITRIC-OXIDE SYNTHASE; SMOOTH-MUSCLE CELLS; RADIATION-THERAPY; PLATELET ACTIVATION; VASCULAR INJURY; INTIMAL INJURY; ANGIOPLASTY; RESTENOSIS; PROLIFERATION AB Purpose: Intracoronary radiation (IR) suppresses neointima formation following balloon injury in animal models. High doses of radiation exacerbate thrombosis and delay re-endothelialization. The free radical nitric oxide (NO) has been reported to inhibit platelet aggregation, reduce neointimal hyperplasia, and stimulate re-endothelialization. This study examined the effects of a chemical NO donor on neointima formation, thrombosis, and healing of irradiated porcine coronary arteries. Methods and Materials: Vascular lesions were created in the coronary arteries of 59 domestic swine by overstretch balloon injury. Arteries were then left untreated or were treated with intracoronary gamma -radiation using Iridium-192 in each artery to deliver 5 or 15 Gy at 2 mm from the center of the source, The chemical NO donor S-nitrosoglutathione (GSNO) was infused i.v. at a rate of 250 mug/min for 10 min before injury, followed by a continuous infusion for 60 min, Animals were euthanized at 14 days and their arteries were analyzed for histomorphometric indices of proliferation and thrombosis. Results: A dose of 15 Gy reduced the ratio of intimal area to medial fracture length (IA/FL) versus control (0.06 +/- 0.05 0.54 +/- 0.10 [p < 0.001]) but increased the nonocclusive thrombosis rate compared to controls (85% vs, 30%; p < 0.05). A low dose of 5 Gy did not affect neointima formation, Treatment with GSNO reduced thrombosis in all treated groups: control, 15%; 5 Gy, 18%; and 15 Gy, 35% (p < 0.05) without affecting neointima formation, Conclusion: Systemic administration of GSNO during balloon injury and IR was tolerated well by the sine and resulted in reduction of the thrombosis rate, especially at high doses, without apparent effect on neointima formation. (C) 2000 Elsevier Science Inc. C1 Washington Hosp Ctr, Inst Cardiovasc Res, Dept Med, Washington, DC 20010 USA. Medstar Res Inst, Washington, DC USA. NCI, Radiat Biol Branch, Bethesda, MD 20892 USA. Washington Hosp Ctr, Dept Radiat Oncol, Washington, DC 20010 USA. RP Waksman, R (reprint author), Washington Hosp Ctr, Inst Cardiovasc Res, Dept Med, 100 Irving St NW,Suite 4B-1, Washington, DC 20010 USA. NR 46 TC 9 Z9 9 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0360-3016 J9 INT J RADIAT ONCOL JI Int. J. Radiat. Oncol. Biol. Phys. PD NOV 1 PY 2000 VL 48 IS 4 BP 1167 EP 1174 DI 10.1016/S0360-3016(00)00730-6 PG 8 WC Oncology; Radiology, Nuclear Medicine & Medical Imaging SC Oncology; Radiology, Nuclear Medicine & Medical Imaging GA 372NB UT WOS:000165238800032 PM 11072176 ER PT J AU Brown, AP Morrissey, RL Tolhurst, TA Crowell, JA Levine, BS AF Brown, AP Morrissey, RL Tolhurst, TA Crowell, JA Levine, BS TI Oral toxicity of 1,2-dithiole-3-thione, a potential cancer chemopreventive agent, in the rat SO INTERNATIONAL JOURNAL OF TOXICOLOGY LA English DT Article DE anemia; cancer chemoprevention; chemopreventive agent; dithiolethione; gastric toxicity; liver ID OLTIPRAZ; DITHIOLETHIONES; PREVENTION; INDUCTION; TRIAL AB This study examined the toxicity of 1,2-dithiole-3-thione (D3T) in rats following 14 days of daily oral (gavage) administration. D3T, an extensive inducer of hepatic phase II drug-metabolizing enzymes, has demonstrated cancer preventive efficacy in rodent models of tumorigenesis and is a candidate drug for cancer prevention. Male and female CD rats (5/sex/dose group) received D3T at dose levels of 0 (corn oil vehicle control), 2, 6, 20, and 60 mg/kg/day. Oral administration of D3T for 14 days at greater than or equal to 20 mg/kg/day resulted in decreased activity, lethargy, rough coat, and piloerection, and toxicologically significant lesions in the stomach, characterized by apoptotic necrosis and hyperplasia of the glandular mucosa. Administration of D3T at 60 mg/kg/day produced anemia in females, decreased body weight gain in males, and increased vacuolation of adrenal cortical cells. Increased liver weights, vacuolation of hepatocytes, and serum chemistry changes, indicative of altered liver function, were observed at greater than or equal to6 mg/kg/day, which were likely due to a pharmacologic effect of D3T on the liver and not considered to be toxicologically significant. Under the conditions of the study, the no-observed-adverse effect level (NOAEL) was 6 mg/kg/day. C1 Univ Illinois, Dept Pharmacol MC 868, Toxicol Res Lab, Chicago, IL 60612 USA. Pathol Associates Int, Chicago, IL USA. Univ Illinois, Dept Pharmaceut, Drug Disposit Res Lab, Chicago, IL USA. NCI, Div Canc Prevent & Control, NIH, Rockville, MD USA. RP Levine, BS (reprint author), Univ Illinois, Dept Pharmacol MC 868, Toxicol Res Lab, Chicago, IL 60612 USA. NR 20 TC 3 Z9 3 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1091-5818 J9 INT J TOXICOL JI Int. J. Toxicol. PD NOV-DEC PY 2000 VL 19 IS 6 BP 375 EP 381 PG 7 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 390GD UT WOS:000166286600001 ER PT J AU Grem, JL AF Grem, JL TI 5-Fluorouracil: forty-plus and still ticking. A review of its preclinical and clinical development SO INVESTIGATIONAL NEW DRUGS LA English DT Review DE antimetabolites; pyrimidine analogs; thymidylate synthase ID COLON-CARCINOMA-CELLS; DIHYDROPYRIMIDINE DEHYDROGENASE-ACTIVITY; ADVANCED COLORECTAL-CANCER; HIGH-DOSE LEUCOVORIN; METASTATIC GASTROINTESTINAL ADENOCARCINOMA; CONTINUOUS-INFUSION 5-FLUOROURACIL; THYMIDYLATE SYNTHASE INHIBITION; BLOOD MONONUCLEAR-CELLS; MOUSE LEUKEMIA-CELLS; PHASE-I TRIAL AB 5-Fluorouracil (5-FU) and 5-fluoro-2'-deoxyuridine (FdUrd) are pyrimidine analogs that have been part of the therapeutic armamentarium for a variety of solid tumors for over forty years. 5-FU has customarily required intravenous administration due to poor and erratic oral bioavailability, while FdUrd has generally been employed for regional administration to the liver or the peritoneal cavity. A great deal of knowledge has been gained concerning the cellular pharmacology and mechanism of action of 5-FU since it was first synthesized in the late 1950's. A more thorough understanding of the factors influencing the metabolic activation of 5-FU and its cellular effects has generated considerable interest in combining it with both modulatory agents such as leucovorin and methotrexate that enhance its metabolism or cytotoxic effects. In addition, 5-FU has also been employed to enhance the therapeutic activity of other antineoplastic agents or modalities such as cisplatin and ionizing radiation with which it can synergize. Appreciation of the clinical pharmacology of 5-FU and FdUrd have led to a variety of schedules that are clinically useful. The preelinical and clinical pharmacology of 5-FU is reviewed to provide a basis for exploring the novel approaches to permit oral administration of 5-FU or its prodrugs that will be described in other articles in this issue. C1 NCI, Cellular & Clin Pharmacol Sect, Dev Therapeut Dept, Med Branch,Natl Naval Med Ctr, Bethesda, MD 20889 USA. RP Grem, JL (reprint author), NCI, Cellular & Clin Pharmacol Sect, Dev Therapeut Dept, Med Branch,Natl Naval Med Ctr, 8901 Wisconsin Ave,Bldg 8,Rm 5101, Bethesda, MD 20889 USA. NR 127 TC 159 Z9 171 U1 2 U2 11 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0167-6997 J9 INVEST NEW DRUG JI Invest. New Drugs PD NOV PY 2000 VL 18 IS 4 BP 299 EP 313 DI 10.1023/A:1006416410198 PG 15 WC Oncology; Pharmacology & Pharmacy SC Oncology; Pharmacology & Pharmacy GA 354UT UT WOS:000089348600002 PM 11081567 ER PT J AU Organisciak, DT Darrow, RM Barsalou, L Kutty, RK Wiggert, B AF Organisciak, DT Darrow, RM Barsalou, L Kutty, RK Wiggert, B TI Circadian-dependent retinal light damage in rats SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article; Proceedings Paper CT Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology CY MAY 11-17, 1997 CL FT LAUDERDALE, FLORIDA SP Assoc Res Vis & Ophthalmol ID ROD OUTER SEGMENT; MESSENGER-RNA; DIURNAL EXPRESSION; PHOTIC INJURY; VISIBLE-LIGHT; CELL-DEATH; IN-VIVO; MELATONIN; RHYTHM; DIMETHYLTHIOUREA AB PURPOSE. TO determine the relative susceptibility of rats to retinal Light damage at different times of the day or night. METHODS. Rats maintained in a dim cyclic light or dark environment were exposed to a single dose of intense green light beginning at various rimes. Normally, Light exposures were for 8 or 3 hours, respectively, although longer and shorter periods were also used. Some animals were treated with the synthetic antioxidant dimethylthiourea (DMTU) before or after the onset of light. The extent of visual cell loss was estimated from measurements of rhodopsin and retinal DNA levels 2 weeks after Light treatment. The time course of retinal DNA fragmentation, and the expression profiles of heme oxygenase-1 (HO-1) and interphotoreceptor retinol binding protein (IRBP) were determined I to 2 days after exposure. RESULTS. When dark-adapted cyclic light-reared or dark-reared rats were exposed to intense Light during normal nighttime hours (2000-0800) the loss of rhodopsin or photoreceptor cell DNA was approximately twofold greater than that found in rats exposed to light during the day (0800-2000). The relative degree of light damage susceptibility persisted in cyclic Light-reared rats after dark adaptation for up to 3 additional days. For rats reared in a reversed light cycle, the light-induced loss of rhodopsin was also reversed. Longer duration Light treatments revealed that dim cyclic light-reared rats were three- to fourfold more susceptible to light damage at 0100 than at 1700 and that dark-reared animals were approximately twofold more susceptible. Intense light exposure at 0100 resulted in greater retinal DNA fragmentation and the earlier appearance of apoptotic DNA ladders than at 1700. The extent of retinal DNA damage also correlated with an induction of retinal HO-I mRNA and with a reduction in IRBP transcription. Antioxidant treatment with DMTU was effective in preventing retinal light damage when given before but not after the onset of Light. CONCLUSIONS. These results confirm earlier work showing greater retinal light damage in rats exposed at night rather than during the day and extend those findings by demonstrating that a single, relatively short, intense Light exposure causes a circadian-dependent, oxidatively induced loss of phororeceptor cells. The light-induced loss of photoreceptor cells is preceded by DNA fragmentation and by alterations in the normal transcriptional events in the retina and within the photoreceptors. The expression profile of an intrinsic retinal factor(s) at the onset of light exposure appears to be important in determining light damage susceptibility. C1 Wright State Univ, Sch Med, Dept Biochem & Mol Biol, Dayton, OH 45435 USA. Wright State Univ, Sch Med, Petticrew Res Lab, Dayton, OH 45435 USA. Wright State Univ, Sch Med, Dept Ophthalmol, Dayton, OH 45435 USA. NEI, Retinal Cell & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Organisciak, DT (reprint author), Wright State Univ, Sch Med, Dept Biochem & Mol Biol, 3640 Colonel Glenn Highway, Dayton, OH 45435 USA. FU NEI NIH HHS [EY-01959] NR 48 TC 91 Z9 94 U1 0 U2 3 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD NOV PY 2000 VL 41 IS 12 BP 3694 EP 3701 PG 8 WC Ophthalmology SC Ophthalmology GA 369LH UT WOS:000165066400005 PM 11053264 ER PT J AU Tran, MT Lausch, RN Oakes, JE AF Tran, MT Lausch, RN Oakes, JE TI Substance P differentially stimulates IL-8 synthesis in human corneal epithelial cells SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID NF-KAPPA-B; HUMAN ASTROCYTOMA-CELLS; ALPHA GENE-EXPRESSION; PROTEIN-KINASE-C; MESSENGER-RNA; MAST-CELLS; PROINFLAMMATORY CYTOKINES; 3'-UNTRANSLATED REGION; INFLAMMATORY CYTOKINES; TACHYKININ RECEPTORS AB PURPOSE. To determine whether substance P (SP), a neuropeptide with proinflammatory properties, specifically interacts with human corneal epithelial cells to stimulate synthesis of the chemokines interleukin (IL)-8, monocyte chemo-attractant protein (MCP)-1, and regulated on activation normal T-cell expressed and secreted (RANTES) protein. METHODS. Primary cultures of human corneal epithelial cells were established from human corneas. Expression of the SP receptor neurokinin (NK)-1 was determined by both the reverse transcription-polymerase chain reaction (RT-PCR) and radiolabeled saturation binding experiments. Synthesis of chemokine-specific RNA in cells stimulated with SP was analyzed by RT-PCR, and quantitation of chemokine protein synthesis was achieved by enzyme-linked immunosorbent assay. RESULTS. Human corneal epithelial cells expressed NK-1 mRNA and bound SP with a K-d characteristic of NK-1. Exposure of cells to SP had no effect on IL-8-specific mRNA synthesis, whereas it increased the half-life of IL-8 transcripts by more than twofold, resulting in significant enhancement of IL-8 synthesis. The capacity of SP to bind to corneal epithelial cells and to induce IL-8 synthesis was abrogated in the presence of a specific NK-1 receptor antagonist. In contrast to IL-8, exposure of cells to SP did not stimulate synthesis of MCP-1 or RANTES. CONCLUSIONS. The results suggest that human corneal cells express NK-1 receptors that specifically bind SP and induce IL-8 synthesis by stabilizing the chemokine's transcripts. C1 Univ S Alabama, Coll Med, Dept Microbiol & Immunol, Mobile, AL 36688 USA. NCI, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Oakes, JE (reprint author), Univ S Alabama, Coll Med, Dept Microbiol & Immunol, MSB 2096, Mobile, AL 36688 USA. FU NEI NIH HHS [EY12713] NR 53 TC 35 Z9 38 U1 0 U2 0 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD NOV PY 2000 VL 41 IS 12 BP 3871 EP 3877 PG 7 WC Ophthalmology SC Ophthalmology GA 369LH UT WOS:000165066400029 PM 11053288 ER PT J AU Lizak, MJ Datiles, MB Aletras, AH Kador, PF Balaban, RS AF Lizak, MJ Datiles, MB Aletras, AH Kador, PF Balaban, RS TI MRI of the human eye using magnetization transfer contrast enhancement SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID DOG AB PURPOSE. To determine the feasibility of using magnetization transfer contrast-enhanced magnetic resonance imaging (MRI) to track cataractous lens changes. METHODS. A fast spin-echo sequence was modified to include a magnetization transfer contrast (MTC) preparation pulse train. This consisted of twenty 8.5-msec sine pulses, 1200 Hz upfield from the water resonance and 1.2-Hz power. The MTC preparation pulse was followed by acquisition through fast spin-echo imaging. The imaging parameters were number of excitations (NEX) = 1, echo time (TE) = 14 msec, recovery time (TR) = 2 sec, echo train length of eight echos, and a matrix size of 256 x 160. To reduce motion artifacts, the volunteers were asked to fixate on a blinking LED. Normal and MTC-enhanced images were acquired from normal volunteers and volunteers with nuclear or cortical cataracts. RESULTS. The eye was adequately imaged, with few motion artifacts appearing. The lens was well resolved, despite the short T-2. The cornea and ciliary body were also clearly visible. In the lens, resolution of the epithelium and cortex were enhanced with MTC. In addition, contrast-to-noise ratios were measured for each image. Examination of the contrast-to-noise ratio confirmed that MTC increased the contrast between the nucleus and cortex. Unenhanced MRIs showed significant differences between the cortex of normal volunteers and volunteers with cataracts. MTC-enhanced images improved the sensitivity to changes in the nucleus. CONCLUSIONS. In this preliminary study, we were able to use MTC-enhanced MRI to obtain high-contrast images of the human lens. Regular and enhanced MRIs detected statistically significant differences between normal and cataractous lenses. C1 NEI, Lab Ocular Therpaeut, NIH, Bethesda, MD 20892 USA. NEI, Ophthalm Genet & Clin Serv Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Lizak, MJ (reprint author), NEI, Lab Ocular Therpaeut, NIH, 10 Ctr Dr,MSC 1850, Bethesda, MD 20892 USA. RI Balaban, Robert/A-7459-2009; OI Balaban, Robert/0000-0003-4086-0948; Datiles, Manuel III B./0000-0003-4660-1664; Aletras, Anthony/0000-0002-3786-3817 NR 8 TC 13 Z9 14 U1 0 U2 1 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD NOV PY 2000 VL 41 IS 12 BP 3878 EP 3881 PG 4 WC Ophthalmology SC Ophthalmology GA 369LH UT WOS:000165066400030 PM 11053289 ER PT J AU Bulterys, M Burchett, SK Culnane, M Cunningham-Schrader, B Dominguez, K Dunkle, L Draper, L Fowler, MG Hanson, C Kpamegan, E Lindegren, ML Martin-Carpenter, L McIntosh, K McNamara, J McSherry, G Mitchell, WG Mofenson, LM Oleske, JM Rhodes, P Shapiro, DE Smith, ME Styrt, B AF Bulterys, M Burchett, SK Culnane, M Cunningham-Schrader, B Dominguez, K Dunkle, L Draper, L Fowler, MG Hanson, C Kpamegan, E Lindegren, ML Martin-Carpenter, L McIntosh, K McNamara, J McSherry, G Mitchell, WG Mofenson, LM Oleske, JM Rhodes, P Shapiro, DE Smith, ME Styrt, B CA Perinatal Safety Review Working Gr TI Nucleoside exposure in the children of HIV-infected women receiving antiretroviral drugs: Absence of clear evidence for mitochondrial disease in children who died before 5 years of age in five United States cohorts SO JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES LA English DT Article DE HIV; mothers; infants; mitochondrial dysfunction; antiretroviral drugs ID INFANT-DEATH-SYNDROME; PERINATAL TRANSMISSION; ZIDOVUDINE TREATMENT; MUSCLE MITOCHONDRIA; LACTIC-ACIDOSIS; RISK-FACTORS; TOXICITY; ANALOGS; SKELETAL; IMPACT AB Background: Nucleoside reverse transcriptase inhibitors (NRTIs) have been associated with mitochondrial toxicity in individuals receiving treatment. A report of two deaths in Europe attributed to mitochondrial dysfunction in HIV-uninfected infants with perinatal NRTI exposure prompted a review of five U.S. cohorts. Methods: Deaths in HIV-exposed children <60 months of age and HIV-uninfected or indeterminate were reviewed. Review included birth history; perinatal antiretroviral drug exposure; hospital, laboratory, and clinic records; death reports; autopsy results; and local physician queries. Deaths were classified as unrelated (Class 1), unlikely related (Class 2), possibly related (Class 3), or highly suggestive or proven relationship (Class 4), to mitochondrial dysfunction; sudden infant death syndrome (SIDS) was categorized separately. Results and Conclusions: Among over 20,000 children of HIV-infected women, over half of whom had been exposed to NRTIs, 223 died. In HIV-uninfected children, 26 deaths were attributed to Class 1, and 4 were attributed to SIDS. In HIV-indeterminate children, 141, 10, 3, and 0 were Classes 1, 2, 3, and 4, respectively; 33 were due to SIDS and 6 could not be classified. There was no indication that antiretroviral exposure was associated with Class 2 or 3 deaths, or deaths from SIDS. A. search for mitochondrial dysfunction among living children in these cohorts is ongoing. C1 Harvard Univ, Childrens Hosp, Sch Med, Div Infect Dis, Boston, MA 02115 USA. US Ctr Dis Control & Prevent, Div HIV AIDS Prevent, Epidemiol Branch, Atlanta, GA USA. NIAID, Div Aids, Pediat Med Branch, NIH, Bethesda, MD 20892 USA. Frontier Sci & Technol Res Fdn Inc, Amherst, NY USA. Bristol Myers Squibb Co, Pharmaceut Res Inst, Wallingford, CT 06492 USA. Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. Clin Trials & Surveys Corp, Baltimore, MD USA. CDC, Div HIV AIDS Prevent Surveillance & Epidemiol, Surveillance Branch, Atlanta, GA 30333 USA. Glaxo Wellcome Res & Dev Ltd, Res Triangle Pk, NC USA. Univ Med & Dent New Jersey, Sch Med, Dept Pediat, Newark, NJ 07103 USA. Univ So Calif, Sch Med, Keck Sch Med, Los Angeles, CA USA. Childrens Hosp Los Angeles, Los Angeles, CA 90027 USA. NICHHD, Pediat Adolescent & Mat AIDS Branch, NIH, Bethesda, MD 20892 USA. CDC, Div HIV AIDS Prevent, Stat & Data Management Branch, Atlanta, GA 30333 USA. Harvard Univ, Sch Publ Hlth, Ctr Biostat AIDS Res, Boston, MA 02115 USA. US FDA, CDER, Div Antiviral Drug Prod, Rockville, MD 20857 USA. RP McIntosh, K (reprint author), Harvard Univ, Childrens Hosp, Sch Med, Div Infect Dis, Enders 609,300 Longwood Ave, Boston, MA 02115 USA. RI Oleske, James/C-1951-2016; OI Oleske, James/0000-0003-2305-5605; Mofenson, Lynne/0000-0002-2818-9808 NR 36 TC 86 Z9 88 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1525-4135 J9 J ACQ IMMUN DEF SYND JI J. Acquir. Immune Defic. Syndr. PD NOV 1 PY 2000 VL 25 IS 3 BP 261 EP 268 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 381DE UT WOS:000165746400008 ER PT J AU Li, R El-Mallakh, RS Herman, MM Kleinman, JE AF Li, R El-Mallakh, RS Herman, MM Kleinman, JE TI Trinucleotide repeat expansion in the beta 1 subunit of the sodium pump in manic-depression illness: a negative study SO JOURNAL OF AFFECTIVE DISORDERS LA English DT Article DE trinucleotide; beta 1 subunit; Na,K-ATPase ID BIPOLAR AFFECTIVE-DISORDER; EXPANDED CAG REPEATS; SCHIZOPHRENIA; ANTICIPATION; PSYCHOSIS AB Background: Trinucleotide repeats have become a recognized molecular abnormality in a variety of neuropsychiatric conditions. Bipolar illness has been purported to be a possible trinucleotide repeat-associated disease. Since abnormalities in the expression and regulation of the sodium- and potassium-activated adenosine triphosphatase (Na,K-ATPase) have been documented in bipolar patients and since the pi subunit of this pump contains a heterogenous GCC repeat, we decided to investigate the possibility of a repeat expansion in beta 1 subunit of Na,K-ATPase in bipolar patients. Methods: DNA from postmortem temporal cerebral cortex tissue of five bipolar subjects and five matched normal controls and five lyumphoblastoid cells lines from the Old Order Amish bipolar pedigrees and match normal controls were used for this study. The GCC rich region of beta 1 DNA was amplified by polymerase chain reaction (PCR) and sequenced. Results: The range of GCC repeat in the beta 1 gene is between 7 and 9 in our population. This is not different in bipolar patients from normal controls. Limitations: This study examined a small number of patients and examined a very limited portion of the locus. Conclusion: It appears that there is not an expansion of the GCC repeat in the beta 1 gene in bipolar patients. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Louisville, Sch Med, Dept Psychiat & Behav Sci, Mood Disorders Res Program, Louisville, KY 40292 USA. NIMH, Ctr Neurosci, Clin Brain Disorders Branch, Washington, DC USA. RP El-Mallakh, RS (reprint author), Univ Louisville, Sch Med, Dept Psychiat & Behav Sci, Mood Disorders Res Program, Louisville, KY 40292 USA. NR 15 TC 5 Z9 5 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-0327 J9 J AFFECT DISORDERS JI J. Affect. Disord. PD NOV PY 2000 VL 60 IS 2 BP 131 EP 136 DI 10.1016/S0165-0327(99)00170-6 PG 6 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 353EU UT WOS:000089262500007 PM 10967372 ER PT J AU Simon-Nobbe, B Probst, G Kajava, AV Oberkofler, H Susani, M Crameri, R Ferreira, F Ebner, C Breitenbach, M AF Simon-Nobbe, B Probst, G Kajava, AV Oberkofler, H Susani, M Crameri, R Ferreira, F Ebner, C Breitenbach, M TI IgE-binding epitopes of enolases, a class of highly conserved fungal allergens SO JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY LA English DT Article DE enolase; structure; epitope; mold; allergy; Cladosporium herbarum; Alternaria alternata; Aspergillus fumigatus; Candida albicans; Saccharomyces cerevisiae ID CANDIDA-ALBICANS; MOLECULAR CHARACTERIZATION; SACCHAROMYCES-CEREVISIAE; CLADOSPORIUM-HERBARUM; ALTERNARIA-ALTERNATA; 2.25-A RESOLUTION; MAJOR ALLERGEN; YEAST ENOLASE; ALPHA-ENOLASE; PROTEIN AB Background: Cladosporium herbarum and Alternaria alternata are two of the most prominent fungal species inducing type I allergy. Previously, we have demonstrated that enolase (Cla h 6) is the second most important allergen of C herbarum in terms of frequency of sensitization. Objective: IgE-reactive B-cell epitopes of C herbarum enolase were analyzed, and cross-reactivity between fungal enolases was investigated. Methods: Cla h 6 glutathione-S-transferase fusion peptides were constructed by means of PCR cloning, A alternata enolase (Alt a 5) was isolated by screening a complementary (c)DNA expression library with a C herbarum enolase DNA probe. Results: Mapping of Cla h 6 IgE-binding epitopes identified a peptide with a length of 69 amino acids (peptide 9), which bound IgE from 8 of 8 patients. Analysis of the conformation of peptide 9 revealed that it does not form a compact structure but rather spans the whole length of the protein, with side chains exposed to solvent at 3 locations. Peptide 9 in the context of Escherichia coli glutathione-S-transferase not only binds IgE but also competitively inhibits IgE binding to Alt a 5. This result indicates that the epitope or epitopes on peptide 9 constitute a major cross-reacting epitope or epitopes on the enolases from C herbarum and A alternata in the case of the! one patient tested. Conclusions: We demonstrated that the glycolytic enzyme enolase is an allergen not only in C herbarum but also in A alternata. Additionally, enolase was shown to exhibit high cross-reactivity to other fungal enolases. On the basis of the results presented here, we propose the use of recombinant Cia h 6 or maybe even peptide 9 of Cia h 6 for diagnosis and possibly therapy of mold allergy. C1 Salzburg Univ, Inst Genet & Gen Biol, A-5020 Salzburg, Austria. Univ Vienna, Dept Pathophysiol, Vienna, Austria. Swiss Inst Allergy & Asthma Res, CH-7270 Davos, Switzerland. Adv Biol Syst Inc, Salzburg, Austria. Landeskrankenanstalten Salzburg, Dept Lab Med, Salzburg, Austria. NIH, Ctr Mol Modeling, CIT, Bethesda, MD 20892 USA. Austrian Acad Sci, Inst Mol Biol, A-5020 Salzburg, Austria. RP Simon-Nobbe, B (reprint author), Salzburg Univ, Inst Genet & Gen Biol, Hellbrunnerstr 34, A-5020 Salzburg, Austria. RI Ferreira, Fatima/E-4889-2011; Kajava, Andrey/E-1107-2014 OI Ferreira, Fatima/0000-0003-0989-2335; Kajava, Andrey/0000-0002-2342-6886 NR 30 TC 68 Z9 73 U1 1 U2 5 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0091-6749 J9 J ALLERGY CLIN IMMUN JI J. Allergy Clin. Immunol. PD NOV PY 2000 VL 106 IS 5 BP 887 EP 895 DI 10.1067/mai.2000.110799 PG 9 WC Allergy; Immunology SC Allergy; Immunology GA 417ZK UT WOS:000167865200013 PM 11080711 ER PT J AU Hull, SC Kass, NE AF Hull, SC Kass, NE TI Adults with cystic fibrosis and (in)fertility: How has the health care system responded? SO JOURNAL OF ANDROLOGY LA English DT Editorial Material ID PREGNANCY C1 NHGRI, Dept Bioeth, Bioeth Sect, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Bioeth Inst, Program Law Eth & Hlth, Baltimore, MD USA. RP Hull, SC (reprint author), NHGRI, Dept Bioeth, Bioeth Sect, NIH, 10 Ctr Dr,Room 10-1C118, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z99 HG999999] NR 13 TC 5 Z9 5 U1 0 U2 0 PU AMER SOC ANDROLOGY, INC PI LAWRENCE PA C/O ALLEN PRESS, INC PO BOX 368, LAWRENCE, KS 66044 USA SN 0196-3635 J9 J ANDROL JI J. Androl. PD NOV-DEC PY 2000 VL 21 IS 6 BP 809 EP 813 PG 5 WC Andrology SC Endocrinology & Metabolism GA 368WJ UT WOS:000090140200004 PM 11105906 ER PT J AU Rapoport, JL Inoff-Germain, G Weissman, MM Greenwald, S Narrow, WE Jensen, PS Lahey, BB Canino, G AF Rapoport, JL Inoff-Germain, G Weissman, MM Greenwald, S Narrow, WE Jensen, PS Lahey, BB Canino, G TI Childhood obsessive-compulsive disorder in the NIMH MECA study: Parent versus child identification of cases SO JOURNAL OF ANXIETY DISORDERS LA English DT Article DE obsessive-compulsive disorder; prevalence; children; adolescents; parent report; child report; detection of cases ID AUTOIMMUNE NEUROPSYCHIATRIC DISORDERS; DIAGNOSTIC INTERVIEW SCHEDULE; STREPTOCOCCAL INFECTIONS; PSYCHIATRIC-DISORDERS; COMMUNITY SAMPLE; ADOLESCENTS; PREVALENCE; EPIDEMIOLOGY; RELIABILITY; SYMPTOMS AB Because as many as 50% of obsessive-compulsive disorder (OCD) cases have had onset by age 15, interest in its detection in childhood is strong. Clinical experience indicates that children often try to keep their OCD secret and that parental report may give marked underestimates. The authors examined the prevalence of childhood OCD in the NIMH Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study, a four-site community survey which allowed comparison of both parent and child report of the child's OCD and related symptoms and disorders. OCD cases, based on structured interviews (DISC-2.3 with DSM-III-X criteria) with 1,285 caretaker-child pairs, were identified separately for parent and child (aged 9 through 17) informants from the MECA database. Cases were then examined for demographic characteristics, for obsessive-compulsive symptoms and other diagnoses reported in cases "missed" by one reporter, and for comorbid disorders. Of a total of 35 (2.7%) identified cases, four (0.3%) were identified by the parent and 32 (2.5%) were identified by the child, with only one overlapping case. In general, when OCD cases were "missed" by one reporter, that reporter did not substitute another disorder. These findings support clinical data that children with OCD often hide their illness and underscore the importance of child interviews for its detection. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. Columbia Univ, Coll Phys & Surg, Dept Psychiat, Div Child Psychiat, New York, NY USA. New York State Psychiat Inst, New York, NY 10032 USA. NIMH, Off Director, Bethesda, MD 20892 USA. Univ Chicago, Dept Psychiat, Chicago, IL 60637 USA. Univ Puerto Rico, Sch Med, Behav Sci Res Inst, San Juan, PR 00936 USA. RP Rapoport, JL (reprint author), NIMH, Child Psychiat Branch, Room 3N202,10 Ctr Dr,MSC 1600, Bethesda, MD 20892 USA. OI Jensen, Peter/0000-0003-2387-0650; Weissman, Myrna/0000-0003-3490-3075 FU NIMH NIH HHS [U01 MH46717, U01 MH46718, U01 MH46725, U01 MH46732] NR 33 TC 75 Z9 75 U1 9 U2 14 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0887-6185 J9 J ANXIETY DISORD JI J. Anxiety Disord. PD NOV-DEC PY 2000 VL 14 IS 6 BP 535 EP 548 DI 10.1016/S0887-6185(00)00048-7 PG 14 WC Psychology, Clinical; Psychiatry SC Psychology; Psychiatry GA 388BP UT WOS:000166157800001 PM 11918090 ER PT J AU Karos, M Chang, YC McClelland, CM Clarke, DL Fu, J Wickes, BL Kwon-Chung, KJ AF Karos, M Chang, YC McClelland, CM Clarke, DL Fu, J Wickes, BL Kwon-Chung, KJ TI Mapping of the Cryptococcus neoformans MAT alpha locus: Presence of mating type-specific mitogen-activated protein kinase cascade homologs SO JOURNAL OF BACTERIOLOGY LA English DT Article ID SACCHAROMYCES-CEREVISIAE; SCHIZOPHYLLUM-COMMUNE; RECEPTOR GENE; VIRULENCE; PHEROMONE; CONTAINS; FUNGI; YEAST; TRANSCRIPTION; CONSERVATION AB In this study we investigated the relationship between the MAT alpha locus of Cryptococcus neoformans and several MAT alpha-specific mitogen-activated protein (MAP) kinase signal transduction cascade genes, including STE12 alpha, STE11 alpha, and STE20 alpha. To resolve the location of the genes, we screened a cosmid library of the MAT alpha strain B-4500 (JEC21), which was chosen for the C. neoformans genome project. We isolated several overlapping cosmids spanning a region of about 71 kb covering the entire MAT alpha locus. It was found that STE12 alpha, STE11 alpha, and STE20 alpha are imbedded within the locus rather than closely linked to the locus. Furthermore, three copies of MF alpha, the mating type alpha-pheromone gene, a MAT alpha-specific myosin gene, and a pheromone receptor (CPR alpha) were identified within the locus. We created a physical map, based on the restriction enzyme BamHI, and identified both borders of the MAT alpha locus. The MAT alpha locus of C. neoformans is approximately 50 kb in size and is one of the largest mating type loci reported among fungi with a one-locus, two-allele mating system. C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. Univ Texas, Hlth Sci Ctr, Dept Microbiol, San Antonio, TX 78229 USA. RP Kwon-Chung, KJ (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10,Room 11C304, Bethesda, MD 20892 USA. FU NIAID NIH HHS [R29 AI043522, R29AI43522] NR 28 TC 39 Z9 40 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD NOV PY 2000 VL 182 IS 21 BP 6222 EP 6227 DI 10.1128/JB.182.21.6222-6227.2000 PG 6 WC Microbiology SC Microbiology GA 364MQ UT WOS:000089896500036 PM 11029445 ER PT J AU Miller, JC Bono, JL Babb, K El-Hage, N Casjens, S Stevenson, B AF Miller, JC Bono, JL Babb, K El-Hage, N Casjens, S Stevenson, B TI A second allele of eppA in Borrelia burdorferi strain B31 is located on the previously undetected circular plasmid cp9-2 SO JOURNAL OF BACTERIOLOGY LA English DT Article ID LYME-DISEASE SPIROCHETE; OSPC GENE; MOLECULAR-CLONING; ETIOLOGIC AGENT; BURGDORFERI B31; PROTEINS; INFECTIVITY; EXPRESSION; HOMOLOGY; SEQUENCE AB Although sequence analysis of Borrelia burgdorferi isolate B31 was recently declared "complete," we found that cultures of this strain can contain a novel 9-kb circular plasmid, cp9-2. The newly described plasmid contains both sequence similarities with and differences from the previously identified B31 plasmid cp9-1 (formerly cp9). cp9-1 and cp9-2 each encode a unique allele of EppA, a putative membrane protein synthesized by B. burgdorferi during mammalian infection. C1 Univ Kentucky, Albert B Chandler Med Ctr, Coll Med, Dept Microbiol & Immunol, Lexington, KY 40536 USA. NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. Univ Utah, Dept Oncol Sci, Div Mol Biol & Genet, Salt Lake City, UT 84132 USA. RP Stevenson, B (reprint author), Univ Kentucky, Albert B Chandler Med Ctr, Coll Med, Dept Microbiol & Immunol, MS 415, Lexington, KY 40536 USA. FU NIAID NIH HHS [AI44254, R56 AI044254, R01 AI044254] NR 48 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 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD NOV PY 2000 VL 182 IS 21 BP 6254 EP 6258 DI 10.1128/JB.182.21.6254-6258.2000 PG 5 WC Microbiology SC Microbiology GA 364MQ UT WOS:000089896500043 PM 11029452 ER PT J AU Duarte, M Graham, K Daher, A Battisti, PL Bannwarth, S Segeral, E Jeang, KT Gatignol, A AF Duarte, M Graham, K Daher, A Battisti, PL Bannwarth, S Segeral, E Jeang, KT Gatignol, A TI Characterization of TRBP1 and TRBP2 - Stable stem-loop structure at the 5 ' end of TRBP2 mRNA resembles HIV-1 TAR and is not found in its processed pseudogene SO JOURNAL OF BIOMEDICAL SCIENCE LA English DT Article DE TRBP; 5 ' mRNA; pseudogene; HIV LTR ID RNA-BINDING-PROTEIN; POLYMERASE-II; SPLIT GENES; KINASE PKR; CYCLIN T1; MOUSE; TRANSACTIVATION; ACTIVATOR; PROGRAMS; DOMAIN AB TRBP1 and TRBP2 cDNAs have been isolated based on the ability of the protein that they encode to bind HIV-1 TAR RNA. The two cDNAs have different 5' end-termini resulting in 21 additional amino acids for TRBP2 protein compared to TRBP1, The corresponding gene is conserved in mammalian species, By PCR amplification of a human library, we have isolated an additional 22 nucleotides in the 5' end of TRBP2 cDNA, Based on the addition of these 22 new nucleotides, the first 87 nucleotides of TRBP2 mRNA can fold into a stable stem-loop structure that resembles TAR RNA. We have also isolated the DNA sequence that represents the TRBP processed pseudogene. The absence of full alignment between TRBP2 full-length cDNA and this sequence suggests that the stem-loop structure could have prevented a complete reverse transcription during pseudogene formation. Using different antibodies, three forms of TRBP can be identified in primate cells at 40, 43 and 50 kD, suggesting a differential expression from the cDNAs and post-translational modifications. Both TRBP1 and TRBP2 activate the basal and the Tat-activated level of the HIV-1 LTR in human and murine cells. Our data indicate that TRBP proteins act at a level prior to Tat function, TRBP could contribute to improved HIV expression in murine models. Copyright (C) 2000 National Science Council, ROC and S. Karger As. Basel. C1 Lady Davis Inst Med Res, McGill AIDS Ctr, Mol Oncol Grp, Montreal, PQ H3T 1E2, Canada. NIAID, Mol Microbiol Lab, Mol Virol Sect, NIH, Bethesda, MD USA. Inst Cochin Genet Mol, U332, F-75014 Paris, France. Inst Cochin Genet Mol, U529 INSERM, F-75014 Paris, France. RP Gatignol, A (reprint author), Lady Davis Inst Med Res, McGill AIDS Ctr, Mol Oncol Grp, 3755 Cote Ste Catherine, Montreal, PQ H3T 1E2, Canada. RI Jeang, Kuan-Teh/A-2424-2008 NR 43 TC 23 Z9 25 U1 0 U2 1 PU KARGER PI BASEL PA ALLSCHWILERSTRASSE 10, CH-4009 BASEL, SWITZERLAND SN 1021-7770 J9 J BIOMED SCI JI J. Biomed. Sci. PD NOV-DEC PY 2000 VL 7 IS 6 BP 494 EP 506 PG 13 WC Cell Biology; Medicine, Research & Experimental SC Cell Biology; Research & Experimental Medicine GA 373EV UT WOS:000165276200007 PM 11060498 ER PT J AU Li, H Yamada, H Akasaka, K Gronenborn, AM AF Li, H Yamada, H Akasaka, K Gronenborn, AM TI Pressure alters electronic orbital overlap in hydrogen bonds SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE GB1; H-bonds; J coupling; pressure; proteins ID STREPTOCOCCAL PROTEIN-G; IMMUNOGLOBULIN-BINDING DOMAIN; PANCREATIC TRYPSIN-INHIBITOR; SCALAR COUPLINGS; CHEMICAL-SHIFTS; NMR MEASUREMENT; SPECTROSCOPY; (3H)J(NC'); LENGTH; IDENTIFICATION AB Pressure-induced changes in (3h)J(NC)' scalar couplings through hydrogen bonds were investigated in the immunoglobulin binding domain of streptococcal protein G. H-1, N-15 and C-13 triple-resonance NMR spectroscopy coupled with the on-line high pressure cell technique was used to monitor (3h)J(NC)' scalar couplings at 30 and 2000 bar in uniformly labeled N-15 and C-13 protein isotopes. Both increased and decreased (3h)J(NC)' scalar couplings were observed at high pressure. No correlation with secondary structure was apparent. The difference in coupling constants as well as pressure-induced chemical shift data suggests a compaction of the helix ends and an increase of the helix pitch at its center in response to pressure. Our data provides the first direct evidence that the electronic orbital overlap in protein backbone hydrogen bonds is altered by pressure. C1 Kobe Univ, Grad Sch Sci & Technol, Kobe, Hyogo, Japan. Kobe Univ, Fac Sci, Kobe, Hyogo, Japan. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Gronenborn, AM (reprint author), Kobe Univ, Grad Sch Sci & Technol, Kobe, Hyogo, Japan. NR 39 TC 31 Z9 32 U1 1 U2 6 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD NOV PY 2000 VL 18 IS 3 BP 207 EP 216 DI 10.1023/A:1026537609584 PG 10 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA 376YQ UT WOS:000165485200004 PM 11142511 ER PT J AU Chou, JJ Li, SP Bax, A AF Chou, JJ Li, SP Bax, A TI Study of conformational rearrangement and refinement of structural homology models by the use of heteronuclear dipolar couplings SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE calmodulin; dipolar coupling; heteronuclear NMR; homology model; liquid crystal; refinement ID CALCIUM-FREE CALMODULIN; X-RAY STRUCTURES; PEPTIDE COMPLEX; CHEMICAL-SHIFT; NMR; PROTEIN; BINDING; RECOGNITION; RESOLUTION; ANISOTROPY AB For an increasing fraction of proteins whose structures are being studied, sequence homology to known structures permits building of low resolution structural models. It is demonstrated that dipolar couplings, measured in a liquid crystalline medium, not only can validate such structural models, but also refine them. Here, experimental H-1-N-15, H-1(alpha)-C-13(alpha), and C-13'-C-13(alpha) dipolar couplings are shown to decrease the backbone rmsd between various homology models of calmodulin (CaM) and its crystal structure. Starting from a model of the Ca2+-saturated C-terminal domain of CaM, built from the structure of Ca2+-free recoverin on the basis of remote sequence homology, dipolar couplings are used to decrease the rmsd between the model and the crystal structure from 5.0 to 1.25 Angstrom. A better starting model, built from the crystal structure of Ca2+-saturated parvalbumin, decreases in rmsd from 1.25 to 0.93 Angstrom. Similarly, starting from the structure of the Ca2+-ligated CaM N-terminal domain, experimental dipolar couplings measured for the Ca2+-free form decrease the backbone rmsd relative to the refined solution structure of apo-CaM from 4.2 to 1.0 Angstrom. C1 NIDDKD, Chem Phys Lab, Bethesda, MD 20892 USA. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Chou, JJ (reprint author), NIDDKD, Chem Phys Lab, Bldg 2, Bethesda, MD 20892 USA. RI Chou, James/N-9840-2013 NR 40 TC 86 Z9 87 U1 0 U2 8 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD NOV PY 2000 VL 18 IS 3 BP 217 EP 227 DI 10.1023/A:1026563923774 PG 11 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA 376YQ UT WOS:000165485200005 PM 11142512 ER PT J AU Ho, NC Jia, LB Driscoll, CC Gutter, EM Francomano, CA AF Ho, NC Jia, LB Driscoll, CC Gutter, EM Francomano, CA TI A skeletal gene database SO JOURNAL OF BONE AND MINERAL RESEARCH LA English DT Article DE Skeletal Gene Database; Skeletal Genome Anatomy Project (SGAP) AB Systematic organization of documented data coupled with ready accessibility is of great value to research. Catalogs and databases are created specifically to meet this purpose. The Skeletal Gene Database evolves as part of the Skeletal Genome Anatomy Project (SGAP), an ongoing multi-institute collaborative effort, to study the functional genome of bone and other skeletal tissues. The primary objective of the Skeletal Gene Database is to create a contemporary list of skeletal-related genes, offering the following information for each gene: gene name, protein name, cellular function, disease(s) caused by mutation of the corresponding gene, chromosomal location, LocusLink number, gene size, exon/intron numbers, messenger RNA (mRNA) coding region size, protein size/molecular weight, Online Mendelian Inheritance in Man (OMIM) number of the gene, UniGene assignment, and PubMed reference. The database includes genes already known and published in the literature as well as novel genes not yet characterized but known to be expressed in skeletal tissue. It will be posted on the web for easy access and swift referencing. The data will be updated in tempo with current and future research, thereby providing an invaluable service to the scientific community interested in obtaining information on bone-related genes. C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. Univ Calif Berkeley, Dept Integrat Biol, Berkeley, CA 94720 USA. Ohio Univ, Dept Biol Sci, Athens, OH 45701 USA. RP Francomano, CA (reprint author), NHGRI, Med Genet Branch, NIH, Rm 10C101, Bethesda, MD 20892 USA. NR 8 TC 41 Z9 44 U1 0 U2 0 PU AMER SOC BONE & MINERAL RES PI WASHINGTON PA 1200 19TH ST, N W, STE 300, WASHINGTON, DC 20036-2422 USA SN 0884-0431 J9 J BONE MINER RES JI J. Bone Miner. Res. PD NOV PY 2000 VL 15 IS 11 BP 2095 EP 2122 DI 10.1359/jbmr.2000.15.11.2095 PG 28 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 366YQ UT WOS:000090034200004 PM 11092392 ER PT J AU Patel, MS Cole, DEC Smith, JD Hawker, GLA Wong, B Trang, H Vieth, R Meltzer, P Rubin, LA AF Patel, MS Cole, DEC Smith, JD Hawker, GLA Wong, B Trang, H Vieth, R Meltzer, P Rubin, LA TI Alleles of the estrogen receptor alpha-gene and an estrogen receptor cotranscriptional activator gene, amplified in breast cancer-1 (AIB1), are associated with quantitative calcaneal ultrasound SO JOURNAL OF BONE AND MINERAL RESEARCH LA English DT Article DE osteoporosis; genetics; ultrasound; calcaneus; estrogen receptor; AIB1 ID BONE-MINERAL-DENSITY; VITAMIN-D-RECEPTOR; POSTMENOPAUSAL WOMEN; MASS ACCUMULATION; CAUCASIAN WOMEN; ELDERLY WOMEN; HIP FRACTURE; FEMORAL-NECK; COACTIVATORS; ATTENUATION AB Quantitative bone ultrasound (QUS) has a significant heritable component. Because estrogen is required for attainment of peak bone mass, we studied alleles of two genes, estrogen receptor cr (ERI) and amplified in breast cancer-1 (AIB1), for their association with QUS, In a volunteer sample of 663 white women aged 18-35 years, bone ultrasound attenuation (BUA), Speed of sound (SOS), and heel stiffness index (SI), the latter consisting of the component measures of BUA and SOS, were measured at the right calcaneus by QUS, Subjects were genotyped for the ERI polymorphisms Xba I and Pvu II and for the AIB1 polyglutamine tract polymorphism. In a multiple regression analysis, ERI genotype was an independent predictor of QUS-SI (p = 0.03), Because AIB1 and ERI enhance gene expression in a coordinate manner, we also searched for interactions. A gene-by-gene interaction effect was seen for QUS-SI (p = 0.009), QUS-BUA (p = 0.03), and QUS-SOS (p = 0.004). These remained significant after the inclusion of clinically relevant variables into the final regression model, Overall, these clinical and genetic factors accounted for up to 16% of the variance in peak QUS; the genetic markers alone accounted for 4-7%, This is the first demonstration of specific genetic effects on calcaneal QUS encoded by alleles of genes directly involved in mediating estrogen effects on bone. C1 Hosp Sick Children, Dept Pediat Genet, Toronto, ON M5G 1X8, Canada. Univ Toronto, Dept Med, Toronto, ON, Canada. Univ Toronto, Dept Lab Med & Pathobiol, Toronto, ON, Canada. Toronto Gen Hosp Genet Resp, Toronto, ON, Canada. Datasmith Solut, Brampton, ON, Canada. Sunnybrook & Womens Coll Hlth Sci Ctr, Div Rheumatol, Toronto, ON, Canada. NHGRI, NIH, Bethesda, MD 20892 USA. RP Rubin, LA (reprint author), St Michaels Hosp, 30 Bond St, Toronto, ON M5B 1W8, Canada. NR 44 TC 32 Z9 34 U1 0 U2 0 PU AMER SOC BONE & MINERAL RES PI WASHINGTON PA 1200 19TH ST, N W, STE 300, WASHINGTON, DC 20036-2422 USA SN 0884-0431 J9 J BONE MINER RES JI J. Bone Miner. Res. PD NOV PY 2000 VL 15 IS 11 BP 2231 EP 2239 DI 10.1359/jbmr.2000.15.11.2231 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 366YQ UT WOS:000090034200016 PM 11092404 ER PT J AU Bryan, JT Morasso, MI AF Bryan, JT Morasso, MI TI The Dlx3 protein harbors basic residues required for nuclear localization, transcriptional activity and binding to Msx1 SO JOURNAL OF CELL SCIENCE LA English DT Article DE Dlx3; homeodomain; transactivation; DNA binding; bipartite nuclear localization signal ID HOMEODOMAIN-DNA COMPLEX; MOUSE EPIDERMAL-CELLS; HOMEOBOX GENES; EXPRESSION; TISSUE; DIFFERENTIATION; IDENTIFICATION; GROWTH; SPECIFICITY; REPRESSION AB The murine Dlx3 protein is a putative transcriptional activator that has been implicated during development and differentiation of epithelial tissue. D1x3 contains homeodomain and mutational analysis has revealed two regions, one N-terminal and one C-terminal to the homeodomain, that act as transcriptional activators in a yeast one-hybrid assay. In addition to transactivation, data are presented to demonstrate specific DNA binding and an association between Dlx3 and the Msx1 protein in vitro, Immunohistochemical analysis confirmed coexpression of Dlx3 and Msx1 proteins in the differentiated layers of murine epidermal tissues. Transcription factor function requires nuclear localization. In this study, the intracellular localization of the green fluorescent protein fused to Dlx3 was examined in keratinocytes induced to differentiate by calcium and is shown to localize to the nucleus. A bipartite nuclear localization signal (NLS) was identified by mutational analysis and shown to be sufficient for nuclear localization. This was demonstrated by insertion of the Dlx3 bipartite NLS sequence into a cytoplasmic fusion protein, GFP-keratin 14, which functionally redirected GFP-keratin 14 expression to the nucleus. Further analysis of Dlx3 NLS mutants revealed that the Dlx3 NLS sequences are required for specific DNA binding, transactivation potential and interactions with the Msx1 protein. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. RP Morasso, MI (reprint author), NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. FU NIAMS NIH HHS [Z01 AR041124-06] NR 39 TC 30 Z9 31 U1 0 U2 0 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 NOV PY 2000 VL 113 IS 22 BP 4013 EP 4023 PG 11 WC Cell Biology SC Cell Biology GA 383FJ UT WOS:000165872200014 PM 11058088 ER PT J AU Lampson, MA Racz, A Cushman, SW McGraw, TE AF Lampson, MA Racz, A Cushman, SW McGraw, TE TI Demonstration of insulin-responsive trafficking of GLUT4 and vpTR in fibroblasts SO JOURNAL OF CELL SCIENCE LA English DT Article DE insulin action; trafficking; GLUT4 ID RAT ADIPOSE-CELLS; GLUCOSE-TRANSPORT ACTIVITY; REGULATED MEMBRANE AMINOPEPTIDASE; HAMSTER OVARY CELLS; SKELETAL-MUSCLE; 3T3-L1 CELLS; SUBCELLULAR TRAFFICKING; PLASMA-MEMBRANE; PHOTOLABELING TECHNIQUE; INTERNALIZATION MOTIF AB Insulin-responsive trafficking of the GLUT4 glucose transporter and the insulin-regulated aminopeptidase (IRAP) in adipose and muscle cells is well established. Insulin regulation of GLUT4 trafficking in these cells underlies the role that adipose tissue and muscle play in the maintenance of whole body glucose homeostasis. GLUT4 is expressed in a very limited number of tissues, most highly in adipose and muscle, while IRAP is expressed in many tissues. IRAP's physiological role in any of the tissues in which it is expressed, however, is unknown. The fact that IRAP, which traffics by the same insulin-regulated pathway as GLUT4, is expressed in 'non-insulin responsive' tissues raises the question of whether these other cell types also have a specialized insulin-regulated trafficking pathway, The existence of an insulin-responsive pathway in other cell types would allow regulation of IRAP activity at the plasma membrane as a potentially important physiological function of insulin. To address this question we use reporter molecules for both GLUT4 and IRAP trafficking to measure insulin-stimulated translocation in undifferentiated cells by quantitative fluorescence microscopy, One reporter (vpTR), a chimera between the intracellular domain of IRAP and the extracellular and transmembrane domains of the transferrin receptor, has been previously characterized. The other is a GLUT4 construct with an exofacial HA epitope and a C-terminal GFP, By comparing these reporters to the transferrin receptor, a marker for general endocytic trafficking, we demonstrate the existence of a specialized, insulin-regulated trafficking pathway in two undifferentiated cell types, neither of which normally express GLUT4, The magnitude of translocation in these undifferentiated cells (approximately threefold) is similar to that reported for the translocation of GLUT4 in muscle cells. Thus, undifferentiated cells have the necessary retention and translocation machinery for an insulin response that is large enough to be physiologically important. C1 Cornell Univ, Weill Grad Sch Med Sci, Dept Biochem, New York, NY 10021 USA. Cornell Univ, Weill Grad Sch Med Sci, Program Physiol Biophys & Mol Med, New York, NY 10021 USA. UCSF, Dept Med, San Francisco, CA USA. NIDDK, Expt diabet Metab & Nutr Sect, NIH, Bethesda, MD USA. RP McGraw, TE (reprint author), Cornell Univ, Weill Grad Sch Med Sci, Dept Biochem, New York, NY 10021 USA. FU NIDDK NIH HHS [DK52852] NR 58 TC 66 Z9 70 U1 0 U2 5 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 NOV PY 2000 VL 113 IS 22 BP 4065 EP 4076 PG 12 WC Cell Biology SC Cell Biology GA 383FJ UT WOS:000165872200019 PM 11058093 ER PT J AU Puri, PL Sartorelli, V AF Puri, PL Sartorelli, V TI Regulation of muscle regulatory factors by DNA-binding, interacting proteins, and post-transcriptional modifications SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Review ID HELIX-LOOP-HELIX; LARGE T-ANTIGEN; HUMAN RHABDOMYOSARCOMA CELLS; SERUM RESPONSE FACTOR; TPA-INDUCED DIFFERENTIATION; SENESCENT HUMAN FIBROBLASTS; CYCLIN-DEPENDENT KINASE-5; ALPHA-ACTIN PROMOTER; MYOD BASIC REGION; MYOGENIC DIFFERENTIATION AB Skeletal muscle differentiation is influenced by multiple pathways, which regulate the activity of myogenic regulatory factors (MRFs)-the myogenic basic helix-loop-helix proteins and the MEF2-family members-in positive or negative ways. Here we wi II review and discuss the network of signals that regulate MRF function during myocyte proliferation, differentiation, and post-mitotic growth. Elucidating the mechanisms governing muscle-specific transcription will provide important insight in better understanding the embryonic development of muscle at the molecular level and will have important implications in setting out strategies aimed at muscle regeneration. Since the activity of MRFs are compromised in tumors of myogenic derivation-the rhabdomyosarcomas-the studies summarized in this review can provide a useful tool to uncover the molecular basis underlying the formation of these rumors. (C) 2000 Wiley-Liss, Inc. C1 Univ Calif San Diego, Dept Biol, La Jolla, CA 92093 USA. NIAMS, Phys Biol Lab, NIH, Bethesda, MD USA. RP Puri, PL (reprint author), Univ Calif San Diego, Dept Biol, La Jolla, CA 92093 USA. NR 234 TC 209 Z9 215 U1 2 U2 8 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 NOV PY 2000 VL 185 IS 2 BP 155 EP 173 DI 10.1002/1097-4652(200011)185:2<155::AID-JCP1>3.0.CO;2-Z PG 19 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 361LT UT WOS:000089725100001 PM 11025438 ER PT J AU Jung, DW Hecht, D Ho, SW O'Connell, BC Kleinman, HK Hoffman, MP AF Jung, DW Hecht, D Ho, SW O'Connell, BC Kleinman, HK Hoffman, MP TI PKC and ERK1/2 regulate amylase promoter activity during differentiation of a salivary gland cell line SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Article ID GROWTH-FACTOR RECEPTOR; KINASE-C-DELTA; PAROTID ACINAR-CELLS; SIGNAL-TRANSDUCTION; SUBMANDIBULAR-GLAND; TYROSINE PHOSPHORYLATION; PROTEIN-KINASES; GENE-EXPRESSION; EGF RECEPTOR; INTEGRIN AB The addition of transforming growth factor alpha (TGF alpha) to a human submandibular gland cell line (HSG) cultured on basement membrane extract Matrigel, synergistically activates the acinar cell-specific salivary amylase promoter. Signaling through beta 1 integrins and increased phosphorylation of ERK1/2 are involved in the increased promoter activity. Phorbol-12-myristate-13-acetate (PMA) and thapsigargin increase amylase promoter activity, suggesting that phorbol ester and calcium-dependent protein kinase C (PKC) pathways are also involved. The combination of specific inhibitors of PKC and MEK1 inhibits the amylase promoter. Inhibitors of the calcium-dependent PKC isoforms alpha, beta, and gamma decrease the promoter activity; however, PKC beta is not detectable in HSG cells. TGF alpha alters the cellular localization of PKC alpha but not -gamma, suggesting PKC alpha is involved in TGF alpha upregulation of the amylase promoter. Furthermore, rottlerin, a PKC delta-specific inhibitor, increases the promoter activity, suggesting PKC isoforms differentially regulate the amylase promoter. In conclusion, beta 1-integrin and TGF alpha signaling pathways regulate the amylase promoter activity in HSG cells. in response to Matrigel and TGF alpha, the activation of both PKC alpha and phosphorylation of ERK1/2 results in synergistic activation of the amylase promoter. Published 2000 Wiley-Liss, Inc.(dagger) C1 NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD USA. RP Hoffman, MP (reprint author), NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, Bldg 30,Room 430,30 Convent Dr, Bethesda, MD 20892 USA. OI O'Connell, Brian/0000-0003-4529-7664 NR 33 TC 14 Z9 15 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 NOV PY 2000 VL 185 IS 2 BP 215 EP 225 DI 10.1002/1097-4652(200011)185:2<215::AID-JCP6>3.0.CO;2-L PG 11 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 361LT UT WOS:000089725100006 PM 11025443 ER PT J AU Robbins, J AF Robbins, J TI Editorial: New ideas in thyroxine-binding globulin biology SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Editorial Material ID PROTEASE INHIBITORS; HOMOLOGY; LEUKOCYTES; SERPINS; SITE C1 NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. RP Robbins, J (reprint author), NIDDKD, Genet & Biochem Branch, NIH, 9000 Rockville Pike,Bldg 10,Room 6C 201A, Bethesda, MD 20892 USA. NR 20 TC 6 Z9 7 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD NOV PY 2000 VL 85 IS 11 BP 3994 EP 3995 DI 10.1210/jc.85.11.3994 PG 2 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000009 PM 11095420 ER PT J AU Pattanaungkul, S Riggs, BL Yergey, AL Vieira, NE O'Fallon, WM Khosla, S AF Pattanaungkul, S Riggs, BL Yergey, AL Vieira, NE O'Fallon, WM Khosla, S TI Relationship of intestinal calcium absorption to 1,25-dihydroxyvitamin D [1,25(OH)(2)D] levels in young versus elderly women: Evidence for age-related intestinal resistance to 1,25(OH)(2)D action SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID VITAMIN-D METABOLISM; BONE GLA-PROTEIN; PARATHYROID-HORMONE; DIETARY CALCIUM; ADVANCING AGE; D-RECEPTOR; INCREASES; HUMANS; RESORPTION; SECRETION AB Intestinal calcium absorption decreases with aging, but it is unclear whether this is attributable to an age-related intestinal resistance to 1,25-dihydroxyvitamin D [1,25(OH)(2)D] action. Thus, we assessed the in vivo dose response of active intestinal calcium absorption to a broad range of circulating 1,25(OH)(2)D levels in elderly [age (mean +/- SD), 72.5 +/-3.0 yr] vs. young women (age, 28.7 +/- 5.3 yr; n = 20 per group), who were stratified into 5 subgroups: group 1 was given a high calcium intake of 75 mmol/day, suppressing 1,25(OH)(2)D levels; group 2 was even a normal calcium diet of 15-30 mmol/day, representing basal 1,25(OH)(2)D levels; group 3 was given a low-calcium diet of 5 mmol/day to stimulate endogenous 1,25(OH)(2)D production; group 4 was given the low-calcium diet plus 1 mug/day 1,25(OH)(2)D; and group 5 was given a low-calcium diet plus 2 mug/day 1,25(OH)(2)D. After 7 days of diet and/or 1,25(OH)(2)D treatment, fasting fractional calcium absorption (FCA) was assessed by a double-tracer method using stable calcium isotopes. Serum 1,25(OH)(2)D and vitamin D-binding protein levels were measured concurrently, and the free 1,25(OH)(2)D index [molar ratio of 1,25(OH)(2)D to DBP] was calculated. FCA was significantly correlated with the free 1,25(OH)(2)D index in the young (R = 0.63, P = 0.003) but not in the elderly women (R = 0.27, P = 0.25). Moreover, the slope of the relationship between FCA and free 1,25(OH)(2)D index (representing intestinal sensitivity to 1,25(OH)(2)D) was significantly greater in the young (compared with the elderly) women [mean +/- SEM, 0.15 +/- 0.04 (young) vs. 0.03 +/- 0.02, elderly, P = 0.03]. Thus, using an experimental design that allowed us to assess FCA over a wide range of 1,25(OH)(2)D levels, we demonstrate that elderly women have a resistance to 1,25(OH)(2)D action that may contribute to their negative calcium balance, secondary hyperparathyroidism, and bone loss. C1 Mayo Clin & Mayo Fdn, Endocrine Res Unit, Rochester, MN 55905 USA. NICHHD, Sect Metab Anal & Mass Spectrometry, Bethesda, MD 20892 USA. Mayo Clin & Mayo Fdn, Biostat Sect, Rochester, MN 55905 USA. RP Khosla, S (reprint author), Mayo Clin & Mayo Fdn, Endocrine Res Unit, 200 1st St SW,5-194 Joseph, Rochester, MN 55905 USA. EM khosla.sundeep@mayo.edu OI Khosla, Sundeep/0000-0002-2936-4372 FU NCRR NIH HHS [RR-00585]; NIA NIH HHS [AG-04875] NR 37 TC 74 Z9 79 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 NOV PY 2000 VL 85 IS 11 BP 4023 EP 4027 DI 10.1210/jc.85.11.4023 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000016 PM 11095427 ER PT J AU Feuillan, PP Jones, JV Barnes, KM Oerter-Klein, K Cutler, GB AF Feuillan, PP Jones, JV Barnes, KM Oerter-Klein, K Cutler, GB TI Boys with precocious puberty due to hypothalamic hamartoma: Reproductive axis after discontinuation of gonadotropin-releasing hormone analog therapy SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID HISTOLOGY AB Hypothalamic hamartoma is an important cause of precocious puberty in boys. Although the GnRH analogs are known to be effective therapy, there are few studies of the recovery of the pituitary-gonadal aids following long-term treatment. To this end, we studied 11 boys with HH after 8.8 +/- 3.2 yr (range, 4.0-12.6) of treatment with the GnRH agonist D-Trp(6),Prog(9),NEt-LHRH. The patients' levels of LH and FSH, testosterone, testis volume, and body mass index were compared with those of six normal boys in pubertal stage IV-V. We found that the patients' mean +/- SD peak GnRH-stimulated LH and FSH had returned to the normal range by 1 yr after stopping therapy. Whereas testosterone returned to normal levels by 1 yr, the patients' testis volume remained smaller than normal until 2 yr after therapy. Ultrasonography revealed diffuse, punctate, echogenic foci in the testicular parenchyma of two patients; these were first observed during GnRH agonist therapy and persisted unchanged after discontinuation of treatment. Neither of these two patients reported pain or testicular discomfort, no mass or irregularity was detected by manual examination in either patient at any time, and levels of beta -hCG and alpha1-fetoprotein were normal. By 4 yr after therapy, all patients had pubertal stage V pubic hair; their body mass index was not different from that of the normal boys at any time point. The dimensions of the patients' hamartomas did not change during or after therapy, and no patient reported new neurological symptoms or signs suggestive of an enlarging lesion at any time during or after discontinuation of treatment. Two families did report episodes of emotional lability and truancy as the patients reentered puberty after discontinuation of treatment. C1 NIH, Warren G Magnuson Clin Ctr, Dev Endocrinol Branch, Bethesda, MD 20892 USA. RP Feuillan, PP (reprint author), NIH, Warren G Magnuson Clin Ctr, Dev Endocrinol Branch, Bethesda, MD 20892 USA. NR 8 TC 13 Z9 15 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 NOV PY 2000 VL 85 IS 11 BP 4036 EP 4038 DI 10.1210/jc.85.11.4036 PG 3 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000018 PM 11095429 ER PT J AU Goebel, SU Peghini, PL Goldsmith, PK Spiegel, AM Gibril, F Raffeld, M Jensen, RT Serrano, J AF Goebel, SU Peghini, PL Goldsmith, PK Spiegel, AM Gibril, F Raffeld, M Jensen, RT Serrano, J TI Expression of the calcium-sensing receptor in gastrinomas SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID ZOLLINGER-ELLISON-SYNDROME; NEOPLASIA-TYPE-I; EXTRACELLULAR CALCIUM; LONG-TERM; (CA-O(2+))-SENSING RECEPTOR; CA-0(2+)-SENSING RECEPTOR; CA2+-SENSING RECEPTOR; POTENTIAL MEDIATOR; MOLECULAR-CLONING; PROTEIN-KINASE AB Extracellular calcium levels are able to influence the secretion of gastrin by gastrinomas and possibly affect the growth pattern. The molecular mechanisms of these functions are not known. The purpose of the present study was to investigate the presence of the calcium-sensing receptor (CaR) in 10 gastrinomas and determine the extent of expression in the tumors. The amounts of CaR messenger ribonucleic acid in eight tumors were determined by quantitative RT-PCR. Protein expression was analyzed by Western blot and immunohistochemistry using a monoclonal antibody (ADD). CaR messenger ribonucleic acid was detected in all gastrinomas with levels ranging from 0.04-3.16 times the amount of beta -actin transcripts. The Western blot showed a major immunoreactive band at 250 kDa and a minor at 140 kDa, corresponding to the receptor dimer and monomer, respectively. Immunohistochemistry demonstrated variable membranous staining in all gastrinomas and normal pancreatic islets. No staining was observed in the normal liver, lymph node, or exocrine pancreas. We conclude that the CaR is present in all gastrinomas, with expression varying by 80-fold. It probably contributes to the calcium-stimulated gastrin release by gastrinomas. Whether the density of the CaR is a determining factor of the magnitude of this gastrin release or plays a role in regulating the growth pattern of the gastrinoma, as it does in other cells, remains unclear at present. C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. NCI, Hematopathol Sect, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Jensen, RT (reprint author), NIDDKD, Digest Dis Branch, NIH, Bldg 10,Room 9C-103,10 Ctr Dr,MSC 1804, Bethesda, MD 20892 USA. NR 61 TC 37 Z9 37 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD NOV PY 2000 VL 85 IS 11 BP 4131 EP 4137 DI 10.1210/jc.85.11.4131 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000033 PM 11095444 ER PT J AU Vythilingam, M Anderson, GM Owens, MJ Halaszynski, TM Bremner, JD Carpenter, LL Heninger, GR Nemeroff, CB Charney, DS AF Vythilingam, M Anderson, GM Owens, MJ Halaszynski, TM Bremner, JD Carpenter, LL Heninger, GR Nemeroff, CB Charney, DS TI Cerebrospinal fluid corticotropin-releasing hormone in healthy humans: Effects of yohimbine and naloxone SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID POSTTRAUMATIC-STRESS-DISORDER; FACTOR-LIKE IMMUNOREACTIVITY; PITUITARY-ADRENAL AXIS; LOCUS-COERULEUS; PARAVENTRICULAR NUCLEUS; CORTISOL SECRETION; ANXIETY DISORDERS; RAT HYPOTHALAMUS; NOREPINEPHRINE; ACTIVATION AB CRH neurons projecting from the paraventricular nucleus (PVN) of the hypothalamus to the median eminence control hypothalamic-pituitary-adrenal (HPA) axis activity. However, CRH neurons outside the PVN as well as PVN neurons projecting to sites other than the median eminence also contribute to the stress response and may play a role in mood and anxiety disorders. We have attempted to investigate possible noradrenergic and opioid regulation of these non-HPA CRH neurons. We hypothesized that yohimbine (an alpha (2)-adrenergic antagonist) would have stimulatory action on non-HPA CRH neurons, whereas naloxone (a mu -opioid receptor antagonist) would not have this effect. Adult normal volunteers received iv yohimbine (n = 5; 0.4 mug/kg), naloxone (n = 4; 125 mug/kg), or placebo (n = 3; 0.9% saline). Cerebrospinal fluid (CSF) was collected continuously, and concentrations of CSF CRH, CSF norepinephrine (NE), and plasma cortisol were measured. Administration of either yohimbine or naloxone caused significant increases in plasma cortisol concentrations over time. Although yohimbine robustly increased CSF NE levels and appeared to increase CSF CRH levels, these effects were not seen after naloxone or placebo administration. Intraindividual correlations were not observed between the measured concentrations of plasma cortisol and CSF CRH for any of the subjects. The results support the idea that CSF CRH concentrations reflect the activity of non-HPA CRH neurons. Although both yohimbine and naloxone stimulated the HPA axis, only yohimbine appeared to have stimulatory effects on central NE and non-HPA CRH. C1 Yale Univ, Sch Med, Dept Psychiat, New Haven, CT 06519 USA. Yale Univ, Sch Med, Dept Child Psychiat, New Haven, CT 06519 USA. Yale Univ, Sch Med, Dept Anesthesiol, New Haven, CT 06519 USA. Emory Univ, Sch Med, Dept Psychiat & Behav Sci, Atlanta, GA 30322 USA. RP Vythilingam, M (reprint author), NIMH, Mood & Anxiety Disorders Res Program, 9000 Rockville Pike,Bldg 10,Room 4N222, Bethesda, MD 20892 USA. EM Vythim@nih.gov RI Owens, Michael/G-5191-2012 FU NIDA NIH HHS [DA-08705]; NIMH NIH HHS [MH-42088, MH-302929] NR 52 TC 32 Z9 33 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD NOV PY 2000 VL 85 IS 11 BP 4138 EP 4145 DI 10.1210/jc.85.11.4138 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000034 PM 11095445 ER PT J AU Serrano, J Goebel, SU Peghini, PL Lubensky, IA Gibril, F Jensen, RT AF Serrano, J Goebel, SU Peghini, PL Lubensky, IA Gibril, F Jensen, RT TI Alterations in the p16(INK4a)/CDKN2A tumor suppressor gene in gastrinomas SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Review ID ZOLLINGER-ELLISON-SYNDROME; ENDOCRINE NEOPLASIA TYPE-1; HUMAN PITUITARY-TUMORS; SOMATOSTATIN RECEPTOR SCINTIGRAPHY; STRAND CONFORMATION POLYMORPHISM; NON-HODGKINS-LYMPHOMAS; METASTATIC GASTRINOMAS; NEUROENDOCRINE TUMORS; SPORADIC GASTRINOMAS; PROTEIN EXPRESSION AB The p16(INK4a)/CDKN2A gene (p16(INK4a)) is frequently altered by homozygous deletion, mutation, or methylation in many nonendocrine tumors, and these alterations may be predictive of recurrence, tumor growth, or aggressiveness. Whether this is true of neuroendocrine tumors such as gastrinomas is unclear. To address this question we analyzed the gastrinomas from 44 patients for p16(INK4a) gene mutations and correlated the results to the tumor's biological behavior, growth pattern, and aggressiveness. No gastrinomas had mutations of exon 1 or exon 2 of the p16(INK4a) gene, although polymorphisms were found in 54%. No homozygous deletions were found. In 52% of the gastrinomas, hypermethylation of a 5'-CpG island of the p16(INK4a) gene promoter was found. To assess the growth behavior of the gastrinomas, all patients were assessed yearly with at least three conventional imaging studies (computed tomography scan, magnetic resonance imaging, and ultrasound), and since 1994 have been assessed with radionuclide scanning using [(111)In-diethylenetriamine pentaacetic acid,DPhe(1)] octreotide. The mean follow-up was 5.1 +/- 0.4 yr (range, 1.2-11.7). The presence or absence of methylation of the p16(INK4a) gene did not correlate with clinical characteristics of the gastrinoma, biological behavior (gastrin release and basal or maximal acid output), the presence or absence of known prognostic factors (tumor size, gastrinoma location, lymph node metastases, liver metastases, and curability), or growth pattern of the gastrinoma postresection. These results indicate that methylation of the p16(INK4a) gene is the most common gene alteration described to date in gastrinomas. Furthermore, because it is independent of disease stage it is probably an early event in the pathogenesis and because it is independent of the primary gastrinoma location, which is now thought to have different origins, methylation of the p16(INK4a) gene is probably a central process in the molecular pathogenesis of these tumors. C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Jensen, RT (reprint author), NIDDKD, Digest Dis Branch, NIH, Bldg 10,Room 9C-103,10 Ctr Dr,MSC 1804, Bethesda, MD 20892 USA. EM robertj@bdg10.niddk.nih.gov NR 101 TC 75 Z9 75 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 NOV PY 2000 VL 85 IS 11 BP 4146 EP 4156 DI 10.1210/jc.85.11.4146 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000035 PM 11095446 ER PT J AU Garcia-Borreguero, D Wehr, TA Larrosa, O Granizo, JJ Hardwick, D Chrousos, GP Friedman, TC AF Garcia-Borreguero, D Wehr, TA Larrosa, O Granizo, JJ Hardwick, D Chrousos, GP Friedman, TC TI Glucocorticoid replacement is permissive for rapid eye movement sleep and sleep consolidation in patients with adrenal insufficiency SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article; Proceedings Paper CT 15th Congress of the European-Sleep-Research-Society CY SEP, 1998 CL MADRID, SPAIN SP European Sleep Res Soc ID CORTICOTROPIN-RELEASING HORMONE; SLOW-WAVE SLEEP; GROWTH-HORMONE; DELTA-SLEEP; STRESS SYSTEM; CORTISOL; HUMANS; PLASMA; SECRETION; PEPTIDE AB There is a well described temporal relation between hormonal secretion and sleep phase, with hormones of the hypothalamic-pituitary-adrenal (HPA) axis possibly playing a role in determining entry into and duration of different sleep stages. In this study sleep features were studied in primary Addison's patients with undetectable levels of cortisol treated in a double blind, randomized, cross-over fashion with either hydrocortisone or placebo supplementation. We found that REM latency was significantly decreased in Addison's patients when receiving hydrocortisone at bedtime, whereas REM sleep time was increased. There was a trend toward an increase in the percentage of time in REM sleep and the number of REM sleep episodes. Waking time after sleep onset was increased, whereas no differences were observed between the two conditions when total sleep time or specific non-REM sleep parameters were evaluated. Our results suggest that in Addison's patients, cortisol plays a positive, permissive role in REM sleep regulation and may help to consolidate sleep. These effects may be mediated either directly by the central effects of glucocorticoids and/or indirectly through CRH and/or ACTH. C1 Charles R Drew Univ Med & Sci, Univ Calif Sch Med, Dept Med, Div Endocrinol, Los Angeles, CA 90059 USA. NIMH, Clin Psychobiol Branch, NIH, Bethesda, MD 20892 USA. NICHHD, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. Fdn Jimenez Diaz, Dept Neurol, Sleep Disorders Unit, E-28040 Madrid, Spain. Univ Calif Los Angeles, Sch Med, Cedars Sinai Res Inst, Res & Epidemiol Unit, Los Angeles, CA 90048 USA. Univ Calif Los Angeles, Sch Med, Cedars Sinai Res Inst, Dept Med,Div Endocrinol, Los Angeles, CA 90048 USA. RP Friedman, TC (reprint author), Charles R Drew Univ Med & Sci, Univ Calif Sch Med, Dept Med, Div Endocrinol, 1721 E 120th St, Los Angeles, CA 90059 USA. EM friedmant@hotmail.com FU NCRR NIH HHS [U54-RR-14616-01] NR 29 TC 38 Z9 38 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD NOV PY 2000 VL 85 IS 11 BP 4201 EP 4206 DI 10.1210/jc.85.11.4201 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000043 PM 11095454 ER PT J AU Harman, SM Metter, EJ Blackman, MR Landis, PK Carter, HB AF Harman, SM Metter, EJ Blackman, MR Landis, PK Carter, HB TI Serum levels of insulin-like growth factor I (IGF-I), IGF-II, IGF-binding protein-3, and prostate-specific antigen as predictors of clinical prostate cancer SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID TUMOR SUPPRESSOR GENE; PEPTIDE ANALOGS; FACTOR AXIS; HYPERPLASIA; CELLS; RECEPTOR; RISK; PLASMA; INSULIN-LIKE-GROWTH-FACTOR-1; RADIOIMMUNOASSAY AB Insulin-like growth factors (IGFs) may play a role in prostate growth, hyperplasia, and malignancy. High plasma IGF-I has been associated with increased prostate cancer risk. In a prospective, cohort, case-control study in the Baltimore Longitudinal Study on Aging population, we examined prostate volume by magnetic resonance imaging, and prostate-specific antigen (PSA), IGF-I, IGF-II, and IGF-binding protein-3 (IGFBP-3) in sera obtained approximately 9 yr before diagnosis of prostate cancer in cases (n = 72) or age-matched controls (n = 127) and in 76 additional Baltimore Longitudinal Study on Aging men (normal subjects) with measured prostate volumes and no prostate cancer. We calculated adjusted odds ratios (OR) by logistic regression, relative risks for significant ORs, and receiver operator curves for prostate cancer, using serum measures alone and in combination. Adjusted ORs for the high vs, low tertile were: for IGF-I, 3.1 [confidence interval (CI), 1.1-8.7]; for IGF-II, 0.2 (CI, 0.07-0.6); for IGFBP-3, 0.71 (CI, 0.3-1.7); and for PSA, 12.5 (CI, 3.8-40.9). For significant ORs, relative risk estimates remained significant at 2.0 for IGF-I, 0.3 for IGF-II, and 5.5 for PSA. Receiver operator curves showed PSA to be the most powerful predictor of prostate cancer. Adding IGF-II to PSA improved prediction. IGF-II was significantly and inversely related (r = -0.219; P < 0.01) and PSA was directly and significantly related (r = 0.461; P < 0.0001) to prostate volume, whereas IGF-I and IBFBP-3 were not. High IGF-I and low IGF-II are independently associated with increased risk of prostate cancer, but PSA level is a much stronger predictor of prostate cancer in the ensuing 10 yr than either IGF-I or IGF-II. The absence of a relationship of IGF-I to prostate size is inconsistent with increased ascertainment in men with large prostates as the source of greater prostate cancer risk associated with IGF-I. Our data suggest that IGF-II may inhibit both prostate growth and development of prostate cancer. C1 NIA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21287 USA. Johns Hopkins Univ, Sch Med, Dept Urol, Baltimore, MD 21287 USA. RP Harman, SM (reprint author), Kronos Res Fdn, 4455 E Camelback Rd,Suite B135, Phoenix, AZ 85018 USA. EM harman@thekronosgroup.com NR 53 TC 164 Z9 166 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 NOV PY 2000 VL 85 IS 11 BP 4258 EP 4265 DI 10.1210/jc.85.11.4258 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000053 PM 11095464 ER PT J AU Stratakis, CA Papageorgiou, T Premkumar, A Pack, S Kirschner, LS Taymans, SE Zhuang, ZP Oelkers, WH Carney, JA AF Stratakis, CA Papageorgiou, T Premkumar, A Pack, S Kirschner, LS Taymans, SE Zhuang, ZP Oelkers, WH Carney, JA TI Ovarian lesions in Carney complex: Clinical genetics and possible predisposition to malignancy SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID PEUTZ-JEGHERS-SYNDROME; SPOTTY SKIN PIGMENTATION; SEX CORD TUMOR; ENDOCRINE OVERACTIVITY; ADENOMA-MALIGNUM; ANNULAR TUBULES; ALLELE LOSS; MYXOMAS; CANCER; SCHWANNOMAS AB Carney complex (CNC) is a familial multiple neoplasia and lentiginosis syndrome (OMIM 160980, http ://www.ncbi.nlm.nih.gov/omim) with features overlapping those of other multiple endocrine neoplasias and hamartomatoses, Peutz-Jeghers syndrome (PJS) in particular. Although a number of patients with CNC and ovarian tumors have been described in individual patient reports, it is unclear whether ovarian lesions constitute a component of the syndrome or are coincidental events. We investigated 18 women with CNC [age at first evaluation, 31.3 +/- 12.1 yr (mean +/- SD)] prospectively for the development of ovarian tumors over a period of 35.7 +/- 30.6 months by physical examination and pelvic ultrasonography. They were compared with 11 women (age at first evaluation, 32.9 +/- 17 yr) who were enrolled under the same protocol (follow up, 32.3 +/- 25.1 months) and served as a control group. In addition, a registry of 178 women from among a total of 309 patients with CNC was searched retrospectively for any having ovarian tumors. Seven available histological specimens were rereviewed. None of the CNC patients had ovarian tumors analogous to those of PJS. Two patients with CNC in the prospective group developed ovarian tumors and were operated upon. One had bilateral oophorectomy for asynchronous serous cystadenomas. The second patient had a unilateral serous cystadenoma. Resected tumor tissue from both patients was tested for genetic abnormalities of the chromosomal regions to which CNC genetic loci have been mapped. Both showed genomic amplification of chromosomal region 2p16. An additional 10 patients had at least 1 sonogram positive for ovarian cysts. Only 1 of the patients in the control group was found to have a persistent, simple ovarian cyst by ultrasonography. The registry of 178 CNC patients included 4 who had undergone surgery for ovarian tumors. The diagnoses included endometrioid adenocarcinoma (1 patient) and metastatic mucinous adenocarcinoma (the primary site was probably ovarian; 1 patient). In addition, 7 of 12 patients (58%) with CNC, who died of other causes, had ovarian lesions at autopsy. In conclusion, although the same stromal tumor, large-cell calcifying Sertoli cell tumor; affects the testes in CNC and PJS, we did not find such tumors in a small population of CNC patients that was studied prospectively or a larger group of CNC patients that was studied retrospectively. The results of our study also suggested that women with CNC commonly develop ovarian cysts and may be at risk for ovarian carcinoma. The chromosome 2p16 CNC locus was involved in ovarian pathology with apparent copy number gain, suggesting that at least molecularly there is some involvement of the CNC gene(s) in these lesions. Although ovarian tumors do not seem to be a major manifestation of CNC, sonography of the ovaries may be part of the initial evaluation for this genetic syndrome in women with CNC; follow-up of any identified lesion is recommended because of the possible risk for malignancy. C1 NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NCI, Dept Diagnost Radiol, Warren G Magnuson Clin Ctr, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Free Univ Berlin, Dept Endocrinol, D-12200 Berlin, Germany. Mayo Clin & Mayo Fdn, Dept Lab Med & Pathol, Rochester, MN 55905 USA. RP Stratakis, CA (reprint author), NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bldg 10,Room 10N262,10 Center Dr,MSC1862, Bethesda, MD 20892 USA. EM stratakc@cc1.nichd.nih.gov RI Pack, Svetlana/C-2020-2014 NR 46 TC 37 Z9 41 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 NOV PY 2000 VL 85 IS 11 BP 4359 EP 4366 DI 10.1210/jc.85.11.4359 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000069 PM 11095480 ER PT J AU Chung, DC Brown, SB Graeme-Cook, F Seto, M Warshaw, AL Jensen, RT Arnold, A AF Chung, DC Brown, SB Graeme-Cook, F Seto, M Warshaw, AL Jensen, RT Arnold, A TI Overexpression of cyclin D1 occurs frequently in human pancreatic endocrine tumors SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID EPIDERMAL GROWTH-FACTOR; SUPPRESSOR GENE; BREAST-CANCER; CELL-CYCLE; NEUROENDOCRINE TUMORS; INCREASED EXPRESSION; ESOPHAGEAL CANCER; TRANSGENIC MICE; PROTEIN; ONCOGENE AB The molecular pathogenesis of human pancreatic endocrine tumors (PETs) is poorly understood. Three independent animal models have pointed to the pivotal role of the G(1)/S cell cycle transition in pancreatic endocrine cell proliferation. We thus hypothesized that the cell cycle regulator cyclin D1 may contribute to the pathogenesis of human PETs. Overexpression of cyclin D1 was identified in 43% of cases, and no correlation was observed with clinical phenotype. The novel observation of frequent overexpression of cyclin D1 suggests that this established oncogene may be implicated in the pathogenesis of human PETs. The absence of detectable alterations in cyclin D1 genomic structure suggests that the mechanism for its oncogenic activation in PETs may be transcriptional or posttranscriptional. C1 Massachusetts Gen Hosp, Gastrointestinal Unit, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Med, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Pathol, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Surg, Boston, MA 02114 USA. Harvard Univ, Sch Med, Boston, MA 02114 USA. Univ Connecticut, Ctr Hlth, Ctr Mol Med, Farmington, CT 06030 USA. Aichi Canc Ctr, Lab Chemotherapy, Nagoya, Aichi 464, Japan. NIH, Digest Dis Branch, Bethesda, MD 20892 USA. RP Chung, DC (reprint author), Massachusetts Gen Hosp, Gastrointestinal Unit, GRJ 825,55 Fruit St, Boston, MA 02114 USA. EM d_chung@helix.mgh.harvard.edu FU NIDDK NIH HHS [DK-01410-11] NR 45 TC 59 Z9 59 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 NOV PY 2000 VL 85 IS 11 BP 4373 EP 4378 DI 10.1210/jc.85.11.4373 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 402ML UT WOS:000166992000071 PM 11095482 ER PT J AU Cooper, GS Ephross, SA Sandler, DP AF Cooper, GS Ephross, SA Sandler, DP TI Menstrual patterns and risk of adult-onset diabetes mellitus SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article; Proceedings Paper CT 32nd Annual Meeting of the Society-for-Epideminologic-Research CY JUN 10-12, 1999 CL BALTIMORE, MARYLAND SP Soc Epideminol Res DE diabetes mellitus (type II); menarche; menstrual cycles; oligomenorrhea; menstrual bleeding; menorrhagia ID WOMEN AB We examined the association between menstrual patterns and risk of developing adult-onset diabetes in a prospective study of 668 white, college-educated women who completed menstrual diaries throughout their reproductive years. We calculated summary measures of cycle length and variability and bleeding duration for ages less than or equal to 22, 23-27, 28-32, and 33-37 years. The analysis included 35,418 person-years of follow-up and 49 self-reported cases of diabetes (median age at diagnosis, 63 years). There was no association between diabetes risk and age at menarche, mean cycle length, cycle variability, or frequency of long cycles (>42 days). Longer bleeding periods in the mid- and late reproductive years were somewhat associated with an increased risk of diabetes (adjusted rate ratio 1.4, 95% confidence interval 1.0-1.8 per day increase in bleeding duration for menses during ages 28-32). These results do not support the association of long or irregular menstrual cycles with post-menopausal diabetes incidence, but do suggest a possible association of longer bleeding duration with subsequent onset of diabetes. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NIEHS, Epidemiol Branch A305, Res Triangle Pk, NC 27709 USA. Glaxo Wellcome Inc, Worldwide Epidemiol Dept, Res Triangle Pk, NC 27709 USA. RP Cooper, GS (reprint author), NIEHS, Epidemiol Branch A305, POB 12233, Res Triangle Pk, NC 27709 USA. OI Sandler, Dale/0000-0002-6776-0018 FU NCRR NIH HHS [M01-RR00064] NR 11 TC 20 Z9 21 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD NOV PY 2000 VL 53 IS 11 BP 1170 EP 1173 DI 10.1016/S0895-4356(00)00240-7 PG 4 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 376TB UT WOS:000165471900013 PM 11106892 ER PT J AU Liu, J Litman, D Rosenberg, MJ Yu, SH Biesecker, LG Weinstein, LS AF Liu, J Litman, D Rosenberg, MJ Yu, SH Biesecker, LG Weinstein, LS TI A GNAS1 imprinting defect in pseudohypoparathyroidism type IB SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID ALBRIGHT HEREDITARY OSTEODYSTROPHY; NUCLEOTIDE REGULATORY PROTEIN; GS-ALPHA GENE; PARATHYROID-HORMONE; DEFICIENT ACTIVITY; MESSENGER-RNA; RESISTANCE; ERYTHROCYTES; MUTATIONS; G(S)ALPHA AB Pseudohypoparathyroidism type IB (PHPIB) is characterized by renal resistance to parathyroid hormone (PTH) and the absence of other endocrine or physical abnormalities. Familial PHPIB has been mapped to 20q13, near GNAS1, which encodes G(s)alpha, the G protein alpha -subunit required for receptor-stimulated cAMP generation. However, G(s)alpha function is normal in blood cells from PHPIB patients, ruling out mutations within the G(s)alpha coding region. In mice G(s)alpha is expressed only from the maternal allele in renal proximal tubules (the site of PTH action) but is biallelically expressed in most other tissues. Studies in patients with Albright hereditary osteodystrophy suggest a similar G(s)alpha imprinting pattern in humans. Here we identify a region upstream of the G(s)alpha promoter that is normally methylated on the maternal allele and unmethylated on the paternal allele, but that is unmethylated on both alleles in all 13 PHPIB patients studied. Within this region is an alternative promoter and first exon (exon 1A), generating transcripts that are normally expressed only from the paternal allele, but that are biallelically expressed in PHPIB patients. Therefore, PHPIB is associated with a paternal-specific imprinting pattern of the exon 1A region on both alleles, which may lead to decreased G(s)alpha expression in renal proximal tubules. We propose that loss of exon 1A imprinting is the cause of PHPIB. C1 NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. NHGRI, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. RP Weinstein, LS (reprint author), NIDDKD, Metab Dis Branch, NIH, Bldg 10,Room 8C101, Bethesda, MD 20892 USA. OI Weinstein, Lee/0000-0002-1899-5152 NR 37 TC 162 Z9 169 U1 0 U2 0 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD NOV PY 2000 VL 106 IS 9 BP 1167 EP 1174 DI 10.1172/JCI10431 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 371PE UT WOS:000165186700012 PM 11067869 ER PT J AU Chao, L Marcus-Samuels, B Mason, MM Moitra, J Vinson, C Arioglu, E Gavrilova, O Reitman, ML AF Chao, L Marcus-Samuels, B Mason, MM Moitra, J Vinson, C Arioglu, E Gavrilova, O Reitman, ML TI Adipose tissue is required for the antidiabetic, but not for the hypolipidemic, effect of thiazolidinediones SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID ACTIVATED RECEPTOR GENE; PPAR-GAMMA; DIABETES-MELLITUS; INSULIN-RESISTANCE; ALPHA-ISOFORM; MICE; TROGLITAZONE; EXPRESSION; OBESITY; DIFFERENTIATION AB There is uncertainty about the site(s) of action of the antidiabetic thiazolidinediones (TZDs). These drugs are agonist ligands of the transcription factor PPAR gamma, which is abundant in adipose tissue but is normally present at very low levels in liver and muscle. We have studied the effects of TZDs in A-ZIP/F-1 mice, which lack white adipose tissue. The A-ZIP/F-1 phenotype strikingly resembles that of humans with severe lipoatrophic diabetes, including the lack of fat, marked insulin resistance and hyperglycemia, hyperlipidemia, and fatty liver. Rosiglitazone or troglitazone treatment did not reduce glucose or insulin levels, suggesting that white adipose tissue is required for the antidiabetic effects of TZDs. However, TZD treatment was effective in lowering circulating triglycerides and increasing whole body fatty acid oxidation in the A-ZIP/F-1 mice, indicating that this effect occurs via targets other than white adipose tissue. A-ZIP/F-1 mice have markedly increased liver PPAR gamma mRNA levels, which may be a general property of fatty livers. Rosiglitazone treatment increased the triglyceride content of the steatotic Livers of A-ZIP/F-1 and ob/ob mice, but not the "lean" livers of fat-transplanted A-ZIP/F-1 mice. In light of this evidence that rosiglitazone acts differently in steatotic livers, the effects of rosiglitazone, particularly on hepatic triglyceride levels, should be examined in humans with hepatic steatosis. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Reitman, ML (reprint author), NIDDKD, Diabet Branch, NIH, Bldg 10,Room 8N-250, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013; OI Reitman, Marc/0000-0002-0426-9475; Oral, Elif/0000-0002-9171-1144 NR 48 TC 264 Z9 273 U1 0 U2 7 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD NOV PY 2000 VL 106 IS 10 BP 1221 EP 1228 DI 10.1172/JCI11245 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 377CD UT WOS:000165493600006 PM 11086023 ER PT J AU Fauci, AS AF Fauci, AS TI Association of American Physicians Presidential Address - The AIDS model: Scientific and policy lessons for the 21st century SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Editorial Material C1 NIAID, NIH, Bethesda, MD 20892 USA. RP Fauci, AS (reprint author), NIAID, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD NOV PY 2000 VL 106 IS 9 BP S23 EP S27 PG 5 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 371PE UT WOS:000165186700019 ER PT J AU Marques, AR Stock, F Gill, V AF Marques, AR Stock, F Gill, V TI Evaluation of a new culture medium for Borrelia burgdorferi SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID POLYMERASE CHAIN-REACTION; EARLY LYME-DISEASE; ERYTHEMA-MIGRANS; REACTION AMPLIFICATION; CEREBROSPINAL-FLUID; SKIN-LESIONS; DIAGNOSIS; DNA; CULTIVATION AB We evaluated the new MPM medium for the growth of Borrelia burgdorferi. All 18 blood samples from 17 patients with Lyme disease were negative. Growth studies showed that by day 4, most organisms in MPM were not viable. Our results reinforce the use of BSK medium as the primary choice for growing B. burgdorferi. C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. NIH, Warren Grant Magnuson Clin Ctr, Clin Pathol Dept, Bethesda, MD 20892 USA. RP Marques, AR (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10,Room 11N228,10 Ctr Dr, Bethesda, MD 20892 USA. NR 18 TC 31 Z9 32 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD NOV PY 2000 VL 38 IS 11 BP 4239 EP 4241 PG 3 WC Microbiology SC Microbiology GA 400VT UT WOS:000166892900055 PM 11060098 ER PT J AU Gutierrez, M Chabner, BA Pearson, D Steinberg, SM Jaffe, ES Cheson, BD Fojo, A Wilson, WH AF Gutierrez, M Chabner, BA Pearson, D Steinberg, SM Jaffe, ES Cheson, BD Fojo, A Wilson, WH TI Role of a doxorubicin-containing regimen in relapsed and resistant lymphomas: An 8-year follow-up study of EPOCH SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID NON-HODGKINS-LYMPHOMA; BONE-MARROW TRANSPLANTATION; P-GLYCOPROTEIN EXPRESSION; CLINICAL DRUG-RESISTANCE; HIGH-DOSE CHEMOTHERAPY; PHASE-II; ETOPOSIDE; VINCRISTINE; TUMORS; TRIAL AB Purpose: Curative up-front regimens for non-Hodgkin's lymphomas contain doxorubicin, vincristine, and cyclophosphamide, whereas salvage regimens generally contain non-cross-resistant agents. We hypothesized that up-front agents may be highly effective for salvage and developed an infusional regimen based on in vitro evidence of increased efficacy. Patients and Methods: A prospective phase II study of etoposide, vincristine, and doxorubicin over 96 hours with bolus cyclophosphamide and oral prednisone (EPOCH) was performed in 131 patients with relapsed or resistant lymphoma. Results: Seventy-nine percent of patients herd aggressive histologies, 46% were considered high risk by the International Prognostic Index, and 34% herd resistant disease. Eighty-eight percent of patients held received at least four of the agents in EPOCH, and 94% had received doxorubicin. In 125 assessable patients, 29 (24%) achieved complete responses and 60 (50%) achieved partial responses. Among 42 patients with resistant disease, 57% responded, and in 28 patients with relapsed aggressive de novo lymphomas, 89% responded with 54% complete responses. With a median follow-up of 76 months, the overall and event-free survivals (EFS) were 17.5 and 7 months, respectively. In 33 patients with sensitive aggressive disease who did not receive stem-cell transplantation, EFS was 19% at 36 months. Toxicity was primarily hematologic, with an 18% incidence of febrile neutropenia. No clinically significant cardiac toxicity was observed, despite no maximum cumulative doxorubicin dose. Conclusion: EPOCH is highly effective in patients who had previously received most/all of the same drugs and produces durable remissions in curable subtypes. Salvage regimens need not contain non-crossresistant agents, and infusional schedules may partially reverse drug resistance and reduce toxicity. (C) 2000 by American Society of Clinical Oncology. C1 NCI, Med Branch, Div Clin Sci, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. RP Wilson, WH (reprint author), NCI, Med Branch, Div Clin Sci, Bldg 10,Room 12N-226,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 34 TC 77 Z9 86 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD NOV 1 PY 2000 VL 18 IS 21 BP 3633 EP 3642 PG 10 WC Oncology SC Oncology GA 370RJ UT WOS:000165135800006 PM 11054436 ER PT J AU Tabak, J Murphey, CR Moore, LE AF Tabak, J Murphey, CR Moore, LE TI Parameter estimation methods for single neuron models SO JOURNAL OF COMPUTATIONAL NEUROSCIENCE LA English DT Article DE model identification; time domain; frequency domain; sensitivity analysis ID MEMBRANE-PROPERTIES; CABLE PROPERTIES; PYRAMIDAL NEURONS; GRANULE CELLS; NMDA AB With the advancement in computer technology, it has become possible to fit complex models to neuronal data. In this work, we test how two methods can estimate parameters of simple neuron models (passive soma) to more complex ones (neuron with one dendritic cylinder and two active conductances). The first method uses classical voltage traces resulting from current pulses injection (time domain), while the second uses measures of the neuron's response to sinusoidal stimuli (frequency domain). Both methods estimate correctly the parameters in all cases studied. However, the time-domain method is slower and more prone to estimation errors in the cable parameters than the frequency-domain method. Because with noisy data the goodness of fit does not distinguish between different solutions, we suggest that running the estimation procedure a large number of times might help find a good solution and can provide information about the interactions between parameters. Also, because the formulation used for the model's response in the frequency domain is analytical, one can derive a local sensitivity analysis for each parameter. This analysis indicates how well a parameter is likely to be estimated and helps choose an optimal stimulation protocol. Finally, the tests suggest a strategy for fitting single-cell models using the two methods examined. C1 Univ Rennes 1, Equipe Neurobiol, CNRS, URA 256, F-35014 Rennes, France. NINDS, Neural Control Lab, NIH, Bethesda, MD 20892 USA. Univ Texas, Med Branch, Dept Physiol & Biophys, Galveston, TX 77550 USA. Univ Paris 05, Lab Neurobiol Reseaux Sensorimoteurs, F-75270 Paris 06, France. RP Tabak, J (reprint author), Univ Rennes 1, Equipe Neurobiol, CNRS, URA 256, F-35014 Rennes, France. EM joel@spine.ninds.nih.gov RI tabak, joel/K-1549-2013 NR 29 TC 16 Z9 16 U1 0 U2 3 PU SPRINGER PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0929-5313 J9 J COMPUT NEUROSCI JI J. Comput. Neurosci. PD NOV PY 2000 VL 9 IS 3 BP 215 EP 236 DI 10.1023/A:1026531603628 PG 22 WC Mathematical & Computational Biology; Neurosciences SC Mathematical & Computational Biology; Neurosciences & Neurology GA 379HV UT WOS:000165637400001 PM 11139040 ER PT J AU O'Fallon, LR Collman, GW Dearry, A AF O'Fallon, LR Collman, GW Dearry, A TI The National Institute of Environmental Health Sciences' research program on children's environmental health SO JOURNAL OF EXPOSURE ANALYSIS AND ENVIRONMENTAL EPIDEMIOLOGY LA English DT Review DE asthma; childhood development; children; community-based research; pesticides; respiratory disease; translational research AB This article highlights the wide array of research programs supported by die National Institute of Environmental Health Sciences (NIEHS) that address issues related to children's environmental health. Special attention is given to the interagency, collaborative Centers for Children's Environmental Health and Disease Prevention Research program. A brief description of each of the eight centers highlights scientific foci and research efforts to date. In addition to discussing NIEHS-supported research programs, the article emphasizes the NIEHS' commitment to the promotion of translating basic research findings into public health knowledge so that culturally sensitive and applicable interventions may be developed. C1 NIEHS, Div Extramural Res & Training, Res Triangle Pk, NC 27709 USA. RP Dearry, A (reprint author), NIEHS, Div Extramural Res & Training, MD EC-21,POB 12233, Res Triangle Pk, NC 27709 USA. NR 11 TC 9 Z9 10 U1 1 U2 2 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1053-4245 J9 J EXPO ANAL ENV EPID JI J. Expo. Anal. Environ. Epidemiol. PD NOV-DEC PY 2000 VL 10 IS 6 BP 630 EP 637 PN 2 PG 8 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 385PY UT WOS:000166013900002 PM 11138655 ER PT J AU Phillips, TM AF Phillips, TM TI Assessing environmental exposure in children: immunotoxicology screening SO JOURNAL OF EXPOSURE ANALYSIS AND ENVIRONMENTAL EPIDEMIOLOGY LA English DT Article DE allergy; cytokines; immune dysregulation; inflammation ID DRIED-BLOOD SPOTS; FILTER-PAPER; CYTOKINES; INSECTICIDES; EXPRESSION; RISK AB Environmental exposure to a number of xenobiotics, including pesticides, can have serious effects on the immune system of children, thus rendering them susceptible to infections or other disease states. To study this problem, a recycling chromatographic system for assessing cytokine profiles in humans has been developed and used for the measurement of immune system function in children with documented exposure to residential pesticides. The system is capable of measuring 30 different analytes in a single sample thus enabling the same time examination of representative markers of immune differentiation and function. In the present study, a cohort of 25 exposed children were examined and shown to exhibit a number of features; all subjects demonstrated some abnormalities in cytokines associated with hematopoiesis. Additionally, elevations in pro-inflammatory cytokines and neuropeptides indicated a state of generalized and neurogenic inflammation. Further analysis indicated that a depression of the cellular arm of the immune system that correlated with clinical indicators of lowered host resistance to infection could also be detected in a subgroup of the exposed subjects. All exposed children demonstrated evidence of hyperstimulation of the humoral immune system as indicated by elevated IL-5 concentrations and clinical allergy. The degree of immune dysregulation in the exposed children was found to be quite marked when compared to similar studies performed on age-matched controls. C1 George Washington Univ, Med Ctr, Immunochem Lab, Washington, DC 20037 USA. RP Phillips, TM (reprint author), NIH, Ultramicro Analyt Immunichem Resource Bioengn & P, ORS, OD, Bldg 13,Room 3N17, Bethesda, MD 20892 USA. NR 28 TC 14 Z9 14 U1 0 U2 5 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1053-4245 J9 J EXPO ANAL ENV EPID JI J. Expo. Anal. Environ. Epidemiol. PD NOV-DEC PY 2000 VL 10 IS 6 BP 769 EP 775 PN 2 PG 7 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 385PY UT WOS:000166013900016 PM 11138669 ER PT J AU Winterfield, JR Swartz, KJ AF Winterfield, JR Swartz, KJ TI A hot spot for the interaction of gating modifier toxins with voltage-dependent ion channels SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Article DE calcium channels; mutagenesis; agatoxin; voltage-sensing domain ID SENSITIVE CALCIUM-CHANNEL; CARDIAC SODIUM-CHANNELS; OMEGA-GRAMMOTOXIN-SIA; ANTHOPLEURIN-A TOXIN; SCORPION TOXIN; K+ CHANNEL; CENTRUROIDES-SCULPTURATUS; RECEPTOR-SITE; DOMAIN-IV; BETA-SUBUNIT AB The gating modifier toxins are a large family of protein toxins that modify either activation or inactivation of voltage-gated ion channels. omega -Aga-IVA is a gating modifier toxin from spider venom that inhibits voltage-gated Ca2+ channels by shifting activation to more depolarized voltages. We identified two Glu residues near the COOH-terminal edge of S3 in the alpha (1A) Ca2+ channel (one in repeat I and the other in repeat IV) that align with Glu residues previously implicated in for-cling the binding sites for gating modifier toxins on K+ and Na+ channels. We found that mutation of the Glu residue in repeat I of the Ca2+ channel had no significant effect on inhibition by omega -Aga-IVA, whereas the equivalent mutation of the Glu in repeat TV disrupted inhibition by the toxin. These results suggest that the COOH-terminal end of 53 within repeat IV contributes to forming a receptor for omega -Aga-IVA. The strong predictive value of previous mapping studies for K+ and Na+ channel toxins argues for a conserved binding motif for gating modifier toxins within the voltage-sensing domains of voltage-gated ion channels. C1 NINDS, Mol Physiol & Biophys Unit, NIH, Bethesda, MD 20892 USA. RP Swartz, KJ (reprint author), NINDS, Mol Physiol & Biophys Unit, NIH, Bldg 36,Room 2C19,36 Convent Dr,MSC 4066, Bethesda, MD 20892 USA. FU Intramural NIH HHS [ZIA NS002945-13] NR 51 TC 76 Z9 80 U1 0 U2 1 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 NOV PY 2000 VL 116 IS 5 BP 637 EP 644 DI 10.1085/jgp.116.5.637 PG 8 WC Physiology SC Physiology GA 372LA UT WOS:000165234100004 PM 11055992 ER PT J AU An, P Luo, H Lu, T O'Brien, SJ Winkler, C AF An, P Luo, H Lu, T O'Brien, SJ Winkler, C TI Genetic heterogeneity and molecular epidemiology of GB virus C hepatitis G virus in China SO JOURNAL OF HUMAN VIROLOGY LA English DT Article DE GB virus C/hepatitis G virus; hepatitis C virus; non-A-E hepatitis; phylogenetic analysis ID A-E HEPATITIS; LIVER-DISEASE; VERTICAL TRANSMISSION; PHYLOGENETIC ANALYSIS; 5'-TERMINAL REGION; BLOOD-DONORS; INFECTION; GENOTYPES; SEQUENCES; IDENTIFICATION AB Objective: The inter- and intrapatient genetic variation of GB virus C (GBV-C)/hepatitis G virus (HGV) was investigated to characterize the molecular epidemiologic profile of GBV-C/HGV infection in China, an area endemic for viral hepatitis. The intrapatient variation of hepatitis C virus (HCV) from the same patients was compared to that of GBV-C/HGV. Study Design/Methods: GB virus C/HGV RNA was amplified by polymerase chain reaction in 88 patients with hepatitis C, hepatitis B or presumed non-A-E hepatitis from three cities in China. Five clones of the GBV-C/HGV NS3 region were sequenced from each GBV-C/HGV RNA-positive patient. The corresponding region of HCV was also sequenced from patients co-infected with HCV. Representative sequences of the GBV-C/HGV NS3 region from each patient and those of isolates from other continents were subjected to phylogenetic analyses. Results: GB virus C/HGV was detected in 22 (25.25%) of 88 patients: 9 (21.4%) of 42 patients with presumed non-A-E hepatitis, 10 (27.7%) of 36 patients with hepatitis C, 3 (30.0%) in 10 patients with hepatitis B and C, and in none of 60 volunteer blood donors. The extent of nucleotide variation was less between Chinese isolates (2.4-17%; median, 10.4%) than between Chinese isolates and seven isolates from outside China (10.5-19.5%; median, 15.3%). Intrapatient sequence variation ranged from 0 to 1.75%, with a mean of 0.57 +/- 0.51%. Phylogenetic analysis grouped most Chinese isolates into four geographically specific clusters with a divergence of 10% to 16% from each other. The ratio of nonsynonymous to synonymous substitutions of GBV-C/HGV (Ka/Ks 0.019) was much lower than for HCV (0.071) in the same patients. Conclusion: Chinese isolates of GBV-C/HGV are genetically distinct. There are local strains as well as shared strains between different locales. The extent of amino acid sequence conservation suggests strong selection against nonsynonymous substitutions in the GBV-C/HGV genome. (C) Lipponcott Williams & Wilkins, Inc. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, Lab Genom Divers, Frederick, MD 21701 USA. Med Coll Ohio, Dept Pharmacol, Toledo, OH 43699 USA. RP Winkler, C (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC, Intramural Res Support Program, Bldg 560,Room 11-85, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 32 TC 0 Z9 0 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1090-9508 J9 J HUMAN VIROL JI J. Human Virol. PD NOV-DEC PY 2000 VL 3 IS 6 BP 299 EP 305 PG 7 WC Infectious Diseases; Virology SC Infectious Diseases; Virology GA 373FT UT WOS:000165278200001 PM 11100910 ER PT J AU Muthukkumar, S Goldstein, J Stein, KE AF Muthukkumar, S Goldstein, J Stein, KE TI The ability of B cells and dendritic cells to present antigen increases during ontogeny SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INFLUENZAE TYPE-B; C CAPSULAR POLYSACCHARIDE; CD4(+) T-CELLS; HEMOPHILUS-INFLUENZAE; CONJUGATE VACCINE; NEISSERIA-MENINGITIDIS; GROUP-A; IMMUNE-RESPONSE; IN-VIVO; CLINICAL-EVALUATION AB The immune response to polysaccharide (PS) Ags in mice is delayed during ontogeny even when administered in a thymus-dependent (TD) form. In this study, Neisseria meningitidis group C IS-tetanus toroid conjugate (MCPS-TT) vaccine was used to examine whether the delay in the development of Ab responses to TD PS conjugate vaccines in neonatal mice is due to defective Ag presentation. The results show that B cells and dendritic cells (DC) from 3- and 7-day-old mice were severely defective in presenting TT and MCPS-TT to Ag-specific T cell clones. The ability of these cells to present Ag reaches adult levels by 4 wk, The development of anti-MCPS and anti-TT Abs in neonatal mice parallels the functional ability of their APC to present Ag. DC from neonatal mice expressed very low levels of MHC class II, costimulatory molecules B7.1, B7.2, and CD11c but high levels of monocyte-specific markers F4/80 and CD11b and granulocyte marker, Ly6G, Significant changes in the expression of these markers were observed as the age of the mice increased. MHC class II, B7.1 and B7.2, and CD11c all increased with age, reaching adult levels between 3 and 3 wk, concurrent with the function of APC, These results demonstrate that one reason neonates fail to produce high titers of anti-PS Abs even when immunized in a TD form is that their B cells and DC are not fully functional. C1 NIH, US FDA, Ctr Biol Evaluat & Res, Div Monoclonal Antibodies, Bethesda, MD 20892 USA. RP Stein, KE (reprint author), NIH, US FDA, Ctr Biol Evaluat & Res, Div Monoclonal Antibodies, Bldg 29B Room 3NN16,29 Lincoln Dr, Bethesda, MD 20892 USA. NR 67 TC 61 Z9 61 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD NOV 1 PY 2000 VL 165 IS 9 BP 4803 EP 4813 PG 11 WC Immunology SC Immunology GA 367RV UT WOS:000090076000007 PM 11046003 ER PT J AU Blazar, BR Lees, CJ Martin, PJ Noelle, RJ Kwon, B Murphy, W Taylor, PA AF Blazar, BR Lees, CJ Martin, PJ Noelle, RJ Kwon, B Murphy, W Taylor, PA TI Host T cells resist graft-versus-host disease mediated by donor leukocyte infusions SO JOURNAL OF IMMUNOLOGY LA English DT Article ID BONE-MARROW TRANSPLANTATION; CHRONIC MYELOGENOUS LEUKEMIA; NATURAL SUPPRESSOR CELLS; CHRONIC MYELOID-LEUKEMIA; CYTOTOXIC LYMPHOCYTES-T; VETO CELLS; ALLOGENEIC MARROW; MONOCLONAL-ANTIBODIES; RADIATION CHIMERAS; IN-VIVO AB Delayed lymphocyte infusions (DLIs) are used to treat relapse occurring post bone marrow transplantation (BMT) and to increase the donor chimerism in recipients receiving nonmyeloablative conditioning. As compared with donor lymphocytes given early post-BMT, DLIs are associated with a reduced risk of graft-vs-host disease (GVHD), The mechanism(s) responsible for such resistance have remained incompletely defined. We now have observed that host T cells present 3 wk after lethal total body irradiation, at the time of DLI, contribute to DLI-GVHD resistance. The infusion of donor splenocytes on day 0, a time when host bone marrow (BM)-derived T cells are absent, results in greater expansion than later post-BMT when host and donor BM-derived T cells coexist, Selective depletion of host T cells with wanti-Thy1 allelic mAb increased the GVHD risk of DLI, indicating that a Thy1(+) host T cell regulated DLI-GVHD lethality. The conditions by which host T cells are required for optimal DLI resistance were determined. Recipients unable to express CD28 or 4-1BB were as susceptible to DLI-GVHD as anti-Thy1 allelic mAb-treated recipients, indicating that CD28 and 4-1BB are critical to DLI-GVHD resistance. Recipients deficient in both perforin and Fas Ligand but not individually were highly susceptible to DLI-GVHD, Recipients that cannot produce IFN-gamma were more susceptible to DLI-GVHD whereas those deficient in IL-12 or p55 TNFRI were not. Collectively, these data indicate that host T cells, which are capable of generating antidonor CTL effector cells, are responsible for the impaired ability of DLI to induce GVHD, These same mechanisms may limit the efficacy of DLI in cancer therapy under some conditions. C1 Univ Minnesota, Ctr Canc, Minneapolis, MN 55455 USA. Dept Pediat, Div Bone Marrow Transplantat, Minneapolis, MN 55455 USA. Fred Hutchinson Canc Res Ctr, Dept Med, Div Clin Res, Seattle, WA 98195 USA. Dartmouth Med Coll, Dept Microbiol, Hanover, NH 03756 USA. Louisiana State Univ, Med Ctr, Dept Ophthalmol, New Orleans, LA 70112 USA. NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Frederick, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Lab Leukocyte Biol, Frederick, MD 21702 USA. RP Blazar, BR (reprint author), Univ Minnesota Hosp, Box 109 Mayo Bldg,420 SE Delaware St, Minneapolis, MN 55455 USA. FU NHLBI NIH HHS [HL-56067, R01 HL-63452]; NIAID NIH HHS [R01 AI-34495] NR 75 TC 61 Z9 65 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD NOV 1 PY 2000 VL 165 IS 9 BP 4901 EP 4909 PG 9 WC Immunology SC Immunology GA 367RV UT WOS:000090076000019 PM 11046015 ER PT J AU Silver, PB Hathcock, KS Chan, CC Wiggert, B Caspi, RR AF Silver, PB Hathcock, KS Chan, CC Wiggert, B Caspi, RR TI Blockade of costimulation through B7/CD28 inhibits experimental autoimmune uveoretinitis, but does not induce long-term tolerance SO JOURNAL OF IMMUNOLOGY LA English DT Article ID T-CELL CLONES; RETINOID-BINDING PROTEIN; IN-VIVO; CD28 COSTIMULATION; NEGATIVE REGULATOR; CTLA-4 ENGAGEMENT; ANTIGEN; ACTIVATION; ANERGY; MOLECULES AB It has been reported that costimulation blockade can result in T cell anergy, We investigated the effects of blocking costimulatory molecules in vivo on the development of experimental autoimmune uveoretinitis (EAU), a model for autoimmune uveitis in humans that is induced in mice by immunization with the retinal Ag interphotoreceptor retinoid binding protein. B10.A mice immunized with a uveitogenic regimen of interphotoreceptor retinoid-binding protein were treated with Abs to B7.1 and B7.2 for 2 wk. Evaluation of EAU and immunological responses 1 wk later showed that disease had been abrogated, and cellular responses were suppressed. To determine whether the costimulation blockade resulted in tolerance, adult-thymectomized mice immunized for uveitis and treated with anti-B7 or anti-CD28 were rechallenged for uveitis induction 5 wk after the initial immunization. Although confirmed to be disease free after the initial immunization, both anti-B7- and anti-CD28-treated mice developed severe EAU and elevated cellular responses after the rechallenge, equivalent to those of control mice. We conclude that in this model costimulatory blockade in vivo prevents the development of autoimmune disease, but does not result in long-term tolerance. The data are compatible with the interpretation that B7/CD28 blockade prevents generation of effector, but not of memory, T cells. C1 NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. NEI, Retinal Cell & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Caspi, RR (reprint author), NEI, Immunol Lab, NIH, Bldg 10,Room 10N 222,10 Ctr Dr, Bethesda, MD 20892 USA. NR 49 TC 41 Z9 44 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 NOV 1 PY 2000 VL 165 IS 9 BP 5041 EP 5047 PG 7 WC Immunology SC Immunology GA 367RV UT WOS:000090076000037 PM 11046033 ER PT J AU Zapata, JM Krajewska, M Krajewski, S Kitada, S Welsh, K Monks, A McCloskey, N Gordon, J Kipps, TJ Gascoyne, RD Shabaik, A Reed, JC AF Zapata, JM Krajewska, M Krajewski, S Kitada, S Welsh, K Monks, A McCloskey, N Gordon, J Kipps, TJ Gascoyne, RD Shabaik, A Reed, JC TI TNFR-associated factor family protein expression in normal tissues and lymphoid malignancies SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NECROSIS-FACTOR RECEPTOR; NF-KAPPA-B; LYMPHOTOXIN-BETA RECEPTOR; TUMOR-CELL LINES; IN-VIVO PATTERNS; TERMINAL KINASE; SIGNALING COMPLEX; IMMUNOHISTOCHEMICAL ANALYSIS; TARGETED DISRUPTION; STRUCTURAL BASIS AB TNFR-associated factors (TRAFs) constitute a family of adapter proteins that associate with particular TNF family receptors, Humans and mice contain six TRAF genes, but little is known about their in vivo expression at the single cell level, The in vivo locations of TRAF1, TRAF2, TRAF5, and TRAF6 were determined in human and mouse tissues by immunohistochemistry. Striking diversity was observed in the patterns of immunostaining obtained for each TRAF family protein, suggesting their expression is independently regulated in a cell type-specific manner, Dynamic regulation of TRAFs was observed in cultured PBLs, where anti-CD3 Abs, mitogenic lectins, and ILs induced marked increases in the steady-state levels of TRAF1, TRAF2, TRAF5, and TRAF6. TRAF1 was also highly inducible by CD40 ligand in cultured germinal center B cells, whereas THAF2, TRAF3, TRAF5, and TRAF6 were relatively unchanged. Analysis of 83 established human tumor cell lines by semiquantitative immunoblotting methods revealed tendencies of certain cancer types to express particular TRAFs, For example, expression of TRAF1 was highly restricted, with B cell lymphomas consistently expressing this TRAF family member. Consistent with results from tumor cell lines, immunohistochemical analysis of 232 non-Hodgkin lymphomas revealed TRAF1 overexpression in 112 (48%) cases, TRAF1 protein levels were also elevated in circulating B cell chronic lymphocytic leukemia specimens (n = 49) compared with normal peripheral blood B cells (p = 0.01), as determined by immunoblotting, These findings contribute to an improved understanding of the cell-specific roles of TRAFs in normal tissues and provide evidence of altered TRAF1 expression in lymphoid malignancies. C1 Burnham Inst, Program Apoptosis & Cell Death Regulat, La Jolla, CA 92037 USA. British Columbia Canc Agcy, Dept Pathol, Vancouver, BC V5Z 4E6, Canada. NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Frederick, MD USA. Univ Birmingham, MRC, Ctr Immune Regulat, Birmingham, W Midlands, England. Univ Calif San Diego, Dept Med Hematol & Oncol, San Diego, CA 92093 USA. Univ Calif San Diego, Dept Pathol, San Diego, CA 92093 USA. RP Reed, JC (reprint author), Burnham Inst, Program Apoptosis & Cell Death Regulat, 10901 N Torrey Pines Rd, La Jolla, CA 92037 USA. RI Zapata, Juan/J-6304-2014; OI Zapata, Juan/0000-0002-0110-0009; Shabaik, Ahmed/0000-0003-1987-3453 FU NCI NIH HHS [CA 69381]; NINDS NIH HHS [NS36821] NR 63 TC 98 Z9 101 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 NOV 1 PY 2000 VL 165 IS 9 BP 5084 EP 5096 PG 13 WC Immunology SC Immunology GA 367RV UT WOS:000090076000043 PM 11046039 ER PT J AU Kirken, RA Erwin, RA Wang, LH Wang, YL Rui, H Farrar, WL AF Kirken, RA Erwin, RA Wang, LH Wang, YL Rui, H Farrar, WL TI Functional uncoupling of the janus kinase 3-Stat5 pathway in malignant growth of human T cell leukemia virus type 1-transformed human T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NF-KAPPA-B; TRANSFORMS RAT FIBROBLASTS; CONSTITUTIVE ACTIVATION; TAX PROTEIN; HTLV-I; V-SRC; SUBSTRATE RECOGNITION; HERPESVIRUS SAIMIRI; STAT3 ACTIVATION; SERINE RESIDUES AB Human T cell leukemia virus type 1 (HTLV-1) transforms cytokine-dependent T lymphocytes and causes adult T cell leukemia, Janus tyrosine kinase (Jak)3 and transcription factors Stat5a and Stat5b are essential for the proliferation of normal T cells and are constitutively hyperactivated in both HTLV-1-transformed human T cell lines and lymphocytes isolated from HTLV-1-infected patients; therefore, a critical role for the Jak3-Stat5 pathway in the progression of this disease has been postulated. We recently reported that tyrphostin AG-490 selectively blocked IL-2 activation of Jak3/Stat5 and growth of murine T cell lines. Here we demonstrate that disruption of Jak3/Stat5a/b signaling with AG-490 (50 muM) blocked the proliferation of primary human T lymphocytes, but paradoxically failed to inhibit the proliferation of HTLV-1-transformed human T cell lines, HuT-102 and MT-2, Structural homologues of AG-490 also inhibited the proliferation of primary human T cells, but not HTLV-1-infected cells. Disruption of constitutive Jak3/Stat5 activation by AG-490 was demonstrated by inhibition of 1) tyrosine phosphorylation of Jak3, Stat5a (Tyr(694)), and Stat5b (Tyr(699)); 2) serine phosphorylation of Stat5a (Ser(726)) as determined by a novel phosphospecific Ab; and 3) Stat5a/b DNA binding to the Stat5-responsive beta -casein promoter. In contrast, AG-490 had no effect on DNA binding by p50/p65 components of NF-kappaB, a transcription factor activated by the HTLV-1-encoded phosphoprotein, Tax. Collectively, these data suggest that the Jak3-Stat5 pathway in HTLV-1-transformed T cells has become functionally redundant for proliferation. Reversal of this functional uncoupling may be required before Jak3/Stat5 inhibitors will be useful in the treatment of this malignancy. C1 Univ Texas, Hlth Sci Ctr, Dept Integrat Biol & Pharmacol, Houston, TX 77030 USA. Sci Applicat Int Corp, IRSP, Frederick, MD USA. NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Div Basic Sci,Cytokine Mol Mech Sect, Frederick, MD 21702 USA. Uniformed Serv Univ Hlth Sci, Dept Pathol, Bethesda, MD 20814 USA. RP Kirken, RA (reprint author), Univ Texas, Hlth Sci Ctr, Dept Integrat Biol, Med Sci Bldg,Room 4-218, Houston, TX 77030 USA. EM rkirken@farmr1.med.uth.tmc.edu NR 55 TC 39 Z9 40 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD NOV 1 PY 2000 VL 165 IS 9 BP 5097 EP 5104 PG 8 WC Immunology SC Immunology GA 367RV UT WOS:000090076000044 PM 11046040 ER PT J AU Bennink, JR AF Bennink, JR TI A novel vaccine approach to stimulate class I restricted T cells SO JOURNAL OF IMMUNOTHERAPY LA English DT Editorial Material ID TOXIC LYMPHOCYTES-T; PROCESSING PATHWAY; EXOGENOUS ANTIGENS; SOLUBLE-PROTEIN; PEPTIDE; RECOGNITION; MOLECULES; VIRUS C1 NIAID, Viral Dis Lab, Bethesda, MD 20892 USA. RP Bennink, JR (reprint author), NIAID, Viral Dis Lab, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 17 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD NOV-DEC PY 2000 VL 23 IS 6 BP 611 EP 612 DI 10.1097/00002371-200011000-00001 PG 2 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 376AF UT WOS:000165435100001 PM 11186148 ER PT J AU Akhyani, N Berti, R Brennan, MB Soldan, SS Eaton, JM McFarland, HF Jacobson, S AF Akhyani, N Berti, R Brennan, MB Soldan, SS Eaton, JM McFarland, HF Jacobson, S TI Tissue distribution and variant characterization of human herpesvirus (HHV)-6: Increased prevalence of HHV-6A in patients with multiple sclerosis SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID POLYMERASE-CHAIN-REACTION; BLOOD MONONUCLEAR-CELLS; PERIPHERAL-BLOOD; MEASLES-VIRUS; LYMPHOTROPIC VIRUS; EXANTHEM-SUBITUM; HEALTHY-ADULTS; DNA; HUMAN-HERPESVIRUS-6; INFECTION AB Human herpesvirus (HHV)-6 has been associated with the pathogenesis of multiple sclerosis (MS) on the basis of serologic, molecular, and histopathologic studies. This study sought to determine the distribution of HHV-6 in different MS body fluids, including serum, saliva, urine, and peripheral blood lymphocytes. The study results extend the observation of an increased frequency of HHV-6 DNA in serum of patients with MS to the unique detection of viral sequences in urine of a subset of patients with MS. Moreover, the HHV-6 identified in these cell-free compartments was predominantly the HHV-6A variant, which has been reported to be neurotropic. These results support the hypothesis that HHV-6 may contribute to the MS disease process. C1 NINDS, Viral Immunol Sect, NIH, Bethesda, MD 20892 USA. George Washington Univ, Dept Genet, Inst Biomed Sci, Washington, DC USA. RP Jacobson, S (reprint author), NINDS, Viral Immunol Sect, NIH, Bldg 10,Rm 5B-16, Bethesda, MD 20892 USA. NR 44 TC 100 Z9 100 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1321 EP 1325 DI 10.1086/315893 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000005 PM 11023456 ER PT J AU Wright, PF Karron, RA Belshe, RB Thompson, J Crowe, JE Boyce, TG Halburnt, LL Reed, GW Whitehead, SS Anderson, EL Wittek, AE Casey, R Eichelberger, M Thumar, B Randolph, VB Udem, SA Chanock, RM Murphy, BR AF Wright, PF Karron, RA Belshe, RB Thompson, J Crowe, JE Boyce, TG Halburnt, LL Reed, GW Whitehead, SS Anderson, EL Wittek, AE Casey, R Eichelberger, M Thumar, B Randolph, VB Udem, SA Chanock, RM Murphy, BR TI Evaluation of a live, cold-passaged, temperature-sensitive, respiratory syncytial virus vaccine candidate in infancy SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT Keystone Symposium on Molecular Approaches to Human Viral Vaccines CY APR, 1999 CL SNOWBIRD, UTAH ID ANTIBODY-RESPONSE; F-GLYCOPROTEIN; SERONEGATIVE CHIMPANZEES; IMMUNE-RESPONSE; VIRAL-INFECTION; YOUNG-CHILDREN; L-GENE; RSV; DISEASE; ADULTS AB A live-attenuated, intranasal respiratory syncytial virus (RSV) candidate vaccine, cpts-248/404, was tested in phase 1 trials in 114 children, including 37 1-2-month-old infants-a target age for RSV vaccines. The cpts-248/404 vaccine was infectious at 104 and 105 plaque-forming units in RSV-naive children and was broadly immunogenic in children >6 months old. Serum and nasal antibody responses in 1-2 month olds were restricted to IgA, had a dominant response to RSV G protein, and had no increase in neutralizing activity. Nevertheless, there was restricted virus shedding on challenge with a second vaccine dose and preliminary evidence for protection from symptomatic disease on natural reexposure. The cpts-248/404 vaccine candidate did not cause fever or lower respiratory tract illness. In the youngest infants, however, cpts-248/404 was unacceptable because of upper respiratory tract congestion associated with Dealt virus recovery. A live attenuated RSV vaccine for the youngest infant will use cpts-248/404 modified by additional attenuating mutations. C1 Vanderbilt Univ, Sch Med, Dept Pediat, Vanderbilt Vaccine Ctr,Div Infect Dis, Nashville, TN 37232 USA. Vanderbilt Univ, Sch Med, Dept Prevent Med, Nashville, TN 37232 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Int Hlth, Ctr Immunizat Res, Baltimore, MD USA. St Louis Univ, Hlth Sci Ctr, Dept Med, Div Infect Dis, St Louis, MO 63103 USA. St Louis Univ, Hlth Sci Ctr, Dept Pediat, Div Infect Dis, St Louis, MO 63103 USA. NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. Wyeth Lederle Vaccines & Pediat, Pearl River, NY USA. RP Wright, PF (reprint author), Vanderbilt Univ, Sch Med, Dept Pediat, Vanderbilt Vaccine Ctr,Div Infect Dis, D7219 MCN, Nashville, TN 37232 USA. RI Crowe, James/B-5549-2009 OI Crowe, James/0000-0002-0049-1079 FU NIAID NIH HHS [AI-000030, AI-000087, AI-15095] NR 47 TC 188 Z9 199 U1 3 U2 9 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1331 EP 1342 DI 10.1086/315859 PG 12 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000007 PM 11010838 ER PT J AU Schooley, RT Spino, C Kuritzkes, D Walker, BD Valentine, FT Hirsch, MS Cooney, E Friedland, G Kundu, S Merigan, TC McElrath, MJ Collier, A Plaeger, S Mitsuyasu, R Kahn, J Haslett, P Uherova, P deGruttola, V Chiu, S Zhang, B Jones, G Bell, D Ketter, N Twadell, T Chernoff, D Rosandich, M AF Schooley, RT Spino, C Kuritzkes, D Walker, BD Valentine, FT Hirsch, MS Cooney, E Friedland, G Kundu, S Merigan, TC McElrath, MJ Collier, A Plaeger, S Mitsuyasu, R Kahn, J Haslett, P Uherova, P deGruttola, V Chiu, S Zhang, B Jones, G Bell, D Ketter, N Twadell, T Chernoff, D Rosandich, M CA AIDS Clin Trials Grp 209 214 Study TI Two double-blinded, randomized, comparative trials of 4 human immunodeficiency virus type 1 (HIV-1) envelope vaccines in HIV-1-infected individuals across a spectrum of disease severity: AIDS clinical trials groups 209 and 214 SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 4th National Conference on Retroviruses and Opportunistic Infections CY JAN 22-26, 1998 CL CHICAGO, ILLINOIS ID CYTOTOXIC T-LYMPHOCYTES; ANTIRETROVIRAL THERAPY; IMMUNE-RESPONSES; IMMUNOLOGICAL RESPONSES; RECOMBINANT GP120; CELL RESPONSES; INFECTION; IMMUNOGENICITY; SAFETY; VIREMIA AB The potential role of human immunodeficiency virus type 1 (HIV-1)-specific immune responses in controlling viral replication in vivo has stimulated interest in enhancing virus-specific immunity by vaccinating infected individuals with HIV-1 or its components, These studies were undertaken to define patient populations most likely to respond to vaccination, with the induction of novel HIV-1-specific cellular immune responses, and to compare the safety and immunogenicity of several candidate recombinant HIV-1 envelope vaccines and adjuvants, New lymphoproliferative responses (LPRs) developed in <30% of vaccine recipients. LPRs were elicited primarily in study participants with a CD4 cell count >350 cells/mm(3) and were usually strain restricted. Responders tended to be more likely than nonresponders to have an undetectable level of HIV-1 RNA at baseline (P = .067), Induction of new cellular immune responses by HIV-1 envelope vaccines is a function of the immunologic stage of disease and baseline plasma HIV-1 RNA level and exhibits considerable vaccine strain specificity. C1 Univ Colorado, Hlth Sci Ctr, Div Infect Dis, Denver, CO 80262 USA. Harvard Univ, Sch Publ Hlth, Stat & Data Anal Ctr, Boston, MA 02115 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. NYU, Sch Med, New York, NY USA. Yale Univ, Sch Med, New Haven, CT USA. Univ Washington, Sch Med, Seattle, WA USA. Stanford Univ, Sch Med, Palo Alto, CA 94304 USA. Univ Calif Los Angeles, Los Angeles, CA USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Genentech Inc, S San Francisco, CA 94080 USA. Chiron Corp, Emeryville, CA 94608 USA. Social & Sci Syst, Rockville, MD USA. NIAID, Div AIDS, NIH, Bethesda, MD 20892 USA. RP Schooley, RT (reprint author), Univ Colorado, Hlth Sci Ctr, Div Infect Dis, B-168,4200 E 9th Ave, Denver, CO 80262 USA. FU NIAID NIH HHS [AI-28568, AI-17665, AI-32770] NR 29 TC 33 Z9 34 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1357 EP 1364 DI 10.1086/315860 PG 8 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000010 PM 11023459 ER PT J AU Gulick, RM Hu, XJ Fiscus, SA Fletcher, CV Haubrich, R Cheng, HL Acosta, E Lagakos, SW Swanstrom, R Freimuth, W Snyder, S Mills, C Fischl, M Pettinelli, C Katzenstein, D AF Gulick, RM Hu, XJ Fiscus, SA Fletcher, CV Haubrich, R Cheng, HL Acosta, E Lagakos, SW Swanstrom, R Freimuth, W Snyder, S Mills, C Fischl, M Pettinelli, C Katzenstein, D CA AIDS Clin Trials Grp Study 359 Pro TI Randomized study of saquinavir with ritonavir or nelfinavir together with delavirdine, adefovir, or both in human immunodeficiency virus-infected adults with virologic failure on indinavir: AIDS Clinical Trials Group study 359 SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 2nd International Workshop on Salvage Therapy for HIV Infection CY MAY 19-21, 1999 CL TORONTO, CANADA ID PROTEASE INHIBITOR THERAPY; ACTIVE ANTIRETROVIRAL THERAPY; COMBINATION THERAPY; CONTAINING REGIMEN; PLUS SAQUINAVIR; HIV-1 INFECTION; FOLLOW-UP; COHORT; ZIDOVUDINE; LAMIVUDINE AB This study compared antiretroviral activity among 6 "salvage" therapy regimens. The study was a prospective, randomized, 2 X 3 factorial, multicenter study of the AIDS Clinical Trials Group. The study enrolled 277 human immunodeficiency virus (HIV)-infected patients naive to nonnucleoside analogues who had taken indinavir >6 months. The patients had 2000-200,000 HIV RNA copies/mL, Patients received saquinavir with ritonavir or nelfinavir together with delavirdine and/or adefovir and were followed for safety and antiretroviral response between baseline and week 16, At week 16, 30% (77/254) of patients had less than or equal to 500 HIV RNA copies/mL, Virologic response did not differ significantly between pooled ritonavir and nelfinavir groups (28% vs. 33%; P = .50) or between pooled delavirdine and delavirdine/adefovir groups (40% vs. 33%; P = .42), Pooled delavirdine groups had a greater virologic response rate than did adefovir groups (40% vs. 18%; P = .002), Overall, one-third of patients who experienced virologic failure on an indinavir-containing regimen suppressed virus load levels while they were taking a new salvage regimen. C1 Cornell Univ, Weill Med Coll, Dept Med, Div Int Med & Infect Dis,Cornell Clin Trials Unit, New York, NY 10021 USA. Harvard Univ, Sch Publ Hlth, Stat & Data Anal Ctr, Boston, MA 02115 USA. Univ N Carolina, Chapel Hill, NC USA. Univ Minnesota, Minneapolis, MN USA. Univ Calif San Diego, San Diego, CA 92103 USA. Stanford Univ, Med Ctr, Palo Alto, CA 94304 USA. Pharmacia & Upjohn Inc, Kalamazoo, MI 49001 USA. Social & Sci Syst, Rockville, MD USA. NIAID, Div AIDS, NIH, Bethesda, MD 20892 USA. Ohio State Univ, Columbus, OH 43210 USA. Univ Miami, Sch Med, Miami, FL USA. RP Gulick, RM (reprint author), Cornell Univ, Weill Med Coll, Dept Med, Div Int Med & Infect Dis,Cornell Clin Trials Unit, Box 566,525 E 68th St, New York, NY 10021 USA. FU NIAID NIH HHS [AI-38858] NR 33 TC 75 Z9 75 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1375 EP 1384 DI 10.1086/315867 PG 10 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000012 PM 11023461 ER PT J AU Lindsey, JC Hughes, MD McKinney, RE Cowles, MK Englund, JA Baker, CJ Burchett, SK Kline, MW Kovacs, A Moye, J AF Lindsey, JC Hughes, MD McKinney, RE Cowles, MK Englund, JA Baker, CJ Burchett, SK Kline, MW Kovacs, A Moye, J TI Treatment-mediated changes in human immunodeficiency virus (HIV) type 1 RNA and CD4 cell counts as predictors of weight growth failure, cognitive decline, and survival in HIV-infected children SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 7th Conference on Retroviruses and Opportunistic Infections CY JAN 29-FEB 03, 2000 CL SAN FRANCISCO, CALIFORNIA ID DISEASE PROGRESSION; VIRAL LOAD; INFANTS; ZIDOVUDINE; THERAPY; LIFE; DIDANOSINE; RISK AB This meta-analysis of 5 large studies of the Pediatric AIDS Clinical Trials Group was undertaken to evaluate the predictive value of antiretroviral treatment-mediated changes in 3 markers of human immunodeficiency virus (HIV) type 1 disease progression-HIV-1 RNA level, CD4 cell count, and CD4 percentage-for weight growth failure, cognitive decline, and survival in HIV-infected children. Proportional hazards models were used to assess the prognostic value of the markers at baseline and after 24 weeks of treatment, with data from 1089 children. Among children receiving nucleoside with or without nonnucleoside reverse-transcriptase inhibitors, higher immunologic and lower virologic markers at baseline and after 24 weeks were significant independent predictors of survival, whereas virologic markers were significant predictors of weight growth and cognitive failure in children >1 year old. The finding of differential age effects on pediatric-specific clinical outcomes emphasizes the need for continued investigation of treatment effects in children. C1 Harvard Univ, Sch Publ Hlth, Ctr Biostat AIDS Res, Boston, MA 02115 USA. Harvard Univ, Sch Publ Hlth, Dept Biostat, Boston, MA 02115 USA. Harvard Univ, Sch Med, Childrens Hosp, Boston, MA USA. Duke Univ, Med Ctr, Pediat AIDS Clin Trials Unit, Durham, NC USA. Univ Iowa, Dept Stat & Actuarial Sci, Iowa City, IA 52242 USA. Baylor Coll Med, Dept Microbiol & Immunol, Houston, TX 77030 USA. Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. Univ So Calif, Med Ctr, Maternal Child Program, Los Angeles, CA USA. NICHHD, NIH, Rockville, MD USA. RP Lindsey, JC (reprint author), Harvard Univ, Sch Publ Hlth, Ctr Biostat AIDS Res, 651 Huntington Ave, Boston, MA 02115 USA. OI moye, john/0000-0001-9976-8586 FU NIAID NIH HHS [AI-41110] NR 27 TC 36 Z9 37 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1385 EP 1393 DI 10.1086/315865 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000013 PM 11010839 ER PT J AU Hoe, NP Kordari, P Cole, R Liu, MY Palzkill, T Huang, WZ McLellan, D Adams, GJ Hu, M Vuopio-Varkila, J Cate, TR Pichichero, ME Edwards, KM Eskola, J Low, DE Musser, JM AF Hoe, NP Kordari, P Cole, R Liu, MY Palzkill, T Huang, WZ McLellan, D Adams, GJ Hu, M Vuopio-Varkila, J Cate, TR Pichichero, ME Edwards, KM Eskola, J Low, DE Musser, JM TI Human immune response to streptococcal inhibitor of complement, a serotype M1 group A Streptococcus extracellular protein involved in epidemics SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 100th Annual Meeting of the American-Society-for-Microbiology CY MAY 21-25, 2000 CL LOS ANGELS, CALIFORNIA SP Amer Soc Microbiol ID GROUP-A STREPTOCOCCUS; TOXIC-SHOCK-SYNDROME; CYSTEINE PROTEASE; GENETIC DIVERSITY; INFECTIONS; PYOGENES; STRAINS; SURFACE; PATHOGENESIS; ANTIBODIES AB Streptococcal inhibitor of complement (Sic) is a highly polymorphic extracellular protein made by serotype M1 group A Streptococcus strains that contributes to bacterial persistence in the mammalian upper respiratory tract. New variants of the Sic protein arise very rapidly by positive selection in human populations during M1 epidemics. The human antibody response to Sic was analyzed, Of 636 persons living in diverse localities, 43% had anti-Sic serum antibodies, but only 16.4% had anti-M1 protein serum antibody. Anti-Sie antibody was also present in nasal wash specimens in high frequency. Linear B cell epitope mapping showed that serum antibodies recognized epitopes located in structurally variable regions of Sic and the amino terminal hypervariable region of the M1 protein. Phage display analyses confirmed that the polymorphic regions of Sic are primary targets of host antibodies. These results support the hypothesis that selection of Sic variants occurs on mucosal surfaces by a mechanism that involves acquired host antibody. C1 NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA. Baylor Coll Med, Dept Microbiol & Immunol, Houston, TX 77030 USA. Baylor Coll Med, Dept Med, Houston, TX 77030 USA. Vet Affairs Res Serv, Memphis, TN USA. Univ Tennessee, Dept Med, Memphis, TN 38104 USA. Vanderbilt Univ, Sch Med, Dept Pediat, Div Infect Dis, Nashville, TN 37212 USA. ID Vaccine Corp, Bothell, WA USA. Univ Rochester, Med Ctr, Dept Microbiol & Immunol, Rochester, NY 14642 USA. Natl Publ Hlth Inst, Dept Bacteriol, Helsinki, Finland. Natl Publ Hlth Inst, Dept Vaccines, Helsinki, Finland. Mt Sinai Hosp, Dept Microbiol, Toronto, ON M5G 1X5, Canada. Univ Toronto, Toronto, ON, Canada. RP Musser, JM (reprint author), NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, 903 S 4th St, Hamilton, MT 59840 USA. RI Low, Donald/B-1726-2012 FU NIAID NIH HHS [AI-33119, AI-45842] NR 42 TC 28 Z9 28 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1425 EP 1436 DI 10.1086/315882 PG 12 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000018 PM 11015234 ER PT J AU Nardin, EH Oliveira, GA Calvo-Calle, JM Castro, ZR Nussenzweig, RS Schmeckpeper, B Hall, BF Diggs, C Bodison, S Edelman, R AF Nardin, EH Oliveira, GA Calvo-Calle, JM Castro, ZR Nussenzweig, RS Schmeckpeper, B Hall, BF Diggs, C Bodison, S Edelman, R TI Synthetic malaria peptide vaccine elicits high levels of antibodies in vaccinees of defined HLA genotypes SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 48th Annual Meeting of the American-Society-of-Tropical-Medicine-and-Hygiene CY NOV 28-DEC 02, 1999 CL WASHINGTON, D.C. SP Amer Soc Trop Med & Hyg ID FALCIPARUM CIRCUMSPOROZOITE PROTEIN; T-CELL EPITOPES; PLASMODIUM-FALCIPARUM; B-CELL; MONOCLONAL-ANTIBODIES; PROTECTIVE IMMUNITY; CONJUGATE VACCINE; MELANOMA PATIENTS; CS-PROTEIN; IMMUNOGENICITY AB A multiple antigen peptide (MAP) malaria vaccine containing minimal Plasmodium falciparum circumsporozoite protein repeat epitopes was assessed for safety and immunogenicity in volunteers of known class II genotypes. The MAP/alum/QS-21 vaccine formulation elicited high levels of parasite-specific antibodies in 10 of 12 volunteers expressing DQB1*0603, DRB1*0401, or DRB1*1101 class II molecules. In contrast, volunteers of other HLA genotypes were low responders or nonresponders. A second study of 7 volunteers confirmed the correlation of class II genotype and high responder phenotype. This is the first demonstration in humans that a peptide vaccine containing minimal T and B cell epitopes composed of only 5 amino acids (N, A, V, D, and P) can elicit antibody titers comparable to multiple exposures to irradiated P falciparum-infected mosquitoes. Moreover, the high-responder phenotypes were predicted by analysis of peptide/HLA interactions in vitro, thus facilitating the rational design of epitope-based peptide vaccines for malaria, as well as for other pathogens. C1 NYU, Sch Med, Dept Med & Mol Parasitol, New York, NY 10010 USA. Johns Hopkins Univ, Immunogenet Labs, Baltimore, MD USA. Univ Maryland, Sch Med, Ctr Vaccine Dev, Baltimore, MD 21201 USA. NIAID, NIH, Bethesda, MD 20892 USA. Univ Maryland, Univ Hlth Ctr, College Pk, MD 20742 USA. USAID, Malaria Vaccine Dev Program, Washington, DC USA. RP Nardin, EH (reprint author), NYU, Sch Med, Dept Med & Mol Parasitol, 341 E 25th St, New York, NY 10010 USA. FU NIAID NIH HHS [AI-25085, N01-AI-45242, N01-AI-45251] NR 40 TC 103 Z9 110 U1 0 U2 5 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1486 EP 1496 DI 10.1086/315871 PG 11 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000026 PM 11023472 ER PT J AU DeHovitz, JA Kovacs, A Feldman, JG Anastos, K Young, M Cohen, M Gange, SJ Melnick, S Greenblatt, RM AF DeHovitz, JA Kovacs, A Feldman, JG Anastos, K Young, M Cohen, M Gange, SJ Melnick, S Greenblatt, RM TI The relationship between virus load response to highly active antiretroviral therapy and change in CD4 cell counts: A report from the women's interagency HIV study SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 37th Annual Meeting of the Infectious-Diseases-Society-of-America CY NOV 18-21, 1999 CL PHILADELPHIA, PENNSYLVANIA SP Infect Dis Soc Amer ID VIROLOGICAL FAILURE; ZIDOVUDINE; LAMIVUDINE; INDINAVIR; INFECTION; ADULTS AB The relationship between the pattern of virus load response to highly active antiretroviral therapy (HAART) and CD4 lymphocyte response was assessed in a cohort of 249 human immunodeficiency virus (HIV) type 1-infected women at 3 times: 1 before and 2 after initiation of therapy, with follow-up of 6-12 months. Patients with a durable response to HAART (i.e >1 log decrease in HIV-1 RNA sustained for the study periods) had a continuous and significant increase in CD4 cell counts over time, whereas those with no response (<0.5 log decrease in HIV-1 RNA) had a sight decline. Patients with a mixed response (initial decrease >1 log, followed by a subsequent decrease <0.5 log) had an increase in CD4 cell count, followed by a plateau. The trend in CD4 cell count differed significantly by response to HAART, with those patients who experienced a durable response having significantly higher CD4 cell counts than others. C1 Suny Downstate Med Ctr, Dept Prevent Med & Community Hlth, Brooklyn, NY 11201 USA. Suny Downstate Med Ctr, Dept Med, Brooklyn, NY 11201 USA. Catholic Med Ctr New York, Dept Med, New York, NY USA. Univ So Calif, Div Pediat Infect Dis, Los Angeles, CA USA. Univ Calif San Francisco, Dept Med, San Francisco, CA USA. Georgetown Univ, Dept Med, Washington, DC USA. Cook Cty Hosp, Dept Med, Chicago, IL 60612 USA. Johns Hopkins Sch Hyg & Publ Hlth, Dept Epidemiol, Baltimore, MD USA. NCI, Analyt Epidemiol Res Branch, Bethesda, MD 20892 USA. RP DeHovitz, JA (reprint author), Suny Downstate Med Ctr, Dept Prevent Med & Community Hlth, 450 Clarkson Ave,Box 1240, Brooklyn, NY 11201 USA. OI Gange, Stephen/0000-0001-7842-512X FU NIAID NIH HHS [UO1-AI-35004, UO1-AI-31834, UO1-AI-34994] NR 15 TC 13 Z9 13 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1527 EP 1530 DI 10.1086/315875 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000033 PM 11010840 ER PT J AU Fernandez, J Levine, OS Sanchez, J Balter, S LaClaire, L Feris, J Romero-Steiner, S AF Fernandez, J Levine, OS Sanchez, J Balter, S LaClaire, L Feris, J Romero-Steiner, S CA Hib Vaccine Evaluation Team TI Prevention of Haemophilus influenzae type b colonization by vaccination: Correlation with serum anti-capsular IgG concentration SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 9th International Conference of Infectious Diseases CY APR, 2000 CL BUENOS AIRES, ARGENTINA ID CONJUGATE VACCINE; PNEUMONIA; CARRIAGE; TRIAL AB Concentrations of serum anti-Haemophilus influenzae type b (anti-Hib) capsular polysaccharide (CPS) greater than or equal to0.15 and greater than or equal to1.0 mug/mL are widely used as surrogates for protection against invasive Hib disease. However, the relationship between serum anti-Hib CPS following immunization and protection against colonization is not known, making it difficult to evaluate new Hib vaccines or combination vaccines. In the Dominican Republic, nasopharyngealswabs were collected from 546 9-month-old infants who had received Hib conjugate vaccine at ages 2, 4, and 6 months and from 600 unvaccinated infants of the same age. The prevalence of Hib colonization was lower among vaccinated infants than among unvaccinated infants (0.9% vs. 2,3%), Among vaccinated infants, protection against colonization was significantly correlated with anti-Hib CPS concentrations greater than or equal to5 mug/mL 1 month following the third dose of vaccine. These results suggest that the concentration of serum anti-Hib CPS needed for protection against colonization is greater than that needed for protection for invasive disease. C1 Clin Infantil Robert Reid Cabral, Santo Domingo, Dominican Rep. Ctr Dis Control & Prevent, Resp Dis Branch, Natl Ctr Infect Dis, Atlanta, GA USA. RP Levine, OS (reprint author), NIAID, Resp Dis Branch, DMID, 6700-B Rockledge Dr,Rm 3255,MSC 7630, Bethesda, MD 20892 USA. OI Romero-Steiner, Sandra/0000-0003-4128-7768 NR 12 TC 52 Z9 54 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1553 EP 1556 DI 10.1086/315870 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000039 PM 11023481 ER PT J AU Biggar, RJ Black, F Whitby, D AF Biggar, RJ Black, F Whitby, D TI Human herpesvirus 8 in Brazilian Amerindians: Does vertical transmission play a role? Reply SO JOURNAL OF INFECTIOUS DISEASES LA English DT Letter ID HUMAN-HERPESVIRUS-8; CHILDREN; ANTIBODIES; INFECTION C1 NCI, Viral Epidemiol Branch, Bethesda, MD 20852 USA. Yale Univ, Sch Publ Hlth, Dept Epidemiol, New Haven, CT USA. RP Biggar, RJ (reprint author), NCI, Viral Epidemiol Branch, 6120 Execut Blvd,Rm 8014, Bethesda, MD 20852 USA. NR 13 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1574 EP 1575 DI 10.1086/315896 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000045 ER PT J AU Trollfors, B Schneerson, R Robbins, JB AF Trollfors, B Schneerson, R Robbins, JB TI Are only antibody levels involved in the protection against pertussis in acellular pertussis vaccine recipients? Reply SO JOURNAL OF INFECTIOUS DISEASES LA English DT Letter ID WHOOPING-COUGH C1 NICHHD, NIH, Bethesda, MD 20892 USA. Goteborg Primary Hlth Care Syst, Goteborg Pertussis Vaccine Study, Gothenburg, Sweden. RP Robbins, JB (reprint author), NICHHD, NIH, Bldg 6,Rm 424, Bethesda, MD 20892 USA. NR 11 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD NOV PY 2000 VL 182 IS 5 BP 1576 EP 1577 DI 10.1086/315900 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 368FA UT WOS:000090106000047 ER PT J AU Fleischmajer, R Kuroda, K Hazan, R Gordon, RE Lebwohl, MG Sapadin, AN Unda, F Iehara, N Yamada, Y AF Fleischmajer, R Kuroda, K Hazan, R Gordon, RE Lebwohl, MG Sapadin, AN Unda, F Iehara, N Yamada, Y TI Basement membrane alterations in psoriasis are accompanied by epidermal overexpression of MMP-2 and its inhibitor TIMP-2 SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE cell adhesion; MMP-2; psoriasis; TIMP-2 ID TRANSFORMING GROWTH FACTOR-BETA-1; TISSUE INHIBITOR; IV COLLAGENASE; MONOCLONAL-ANTIBODIES; METALLOPROTEINASES-2 TIMP-2; MATRIX METALLOPROTEINASE-2; DIFFERENTIAL EXPRESSION; INTEGRIN EXPRESSION; HUMAN FIBROBLASTS; HUMAN SKIN AB Psoriasis is most probably an inherited disease characterized by cell proliferation, angiogenesis, and an inflammatory process. The pathophysiology remains unknown, although an alteration in cell-cell and cell-matrix adhesion versus an autoimmune process has been proposed as the primary defect. Here, we show evidence of a new mechanism involving basement membrane alterations accompanied by keratinocyte overexpression of matrix metalloproteinase (MMP) 2 and tissue inhibitor of MMP-2 (TIMP-2) in both uninvolved and involved psoriatic skin. Immunocytochemistry with antibodies against collagen IV (alpha1, alpha2 chains) and laminins (alpha2, alpha5, beta1, gamma1 chains) revealed gaps, folding, and reduplication of the epidermo-dermal basement membrane. There was overexpression of MMP-2 in the cytoplasm of suprabasal keratinocytes. Gelatin zymography revealed pro-MMP-2 and its activated form, a-MMP-2, in both uninvolved and involved psoriatic skin, whereas pro-MMP-9 was only present in involved skin. TIMP-2 was expressed at the cell surface of psoriatic involved suprabasal keratinocytes whereas it was restricted to basal keratinocytes in uninvolved areas. Western blots showed a marked increase in a-MMP-2 and TIMP-2 in uninvolved and involved psoriatic skin although it was more pronounced in the latter. MT1-MP, known to activate pro-MMP-2, was increased in involved areas. In situ hybridization revealed strong signals of MMP-2 mRNA in both uninvolved and involved psoriatic epidermis. The overexpression of MMP-2 in uninvolved and involved psoriatic epidermis supports the concept that the primary alteration may reside in the keratinocyte. In addition, the presence of the activated form of MMP-2 could be responsible for cell-cell and cell-matrix changes noted in psoriatic epidermis. C1 Mt Sinai Med Ctr, Dept Dermatol, New York, NY 10029 USA. Mt Sinai Med Ctr, Ctr Canc, New York, NY 10029 USA. Mt Sinai Med Ctr, Dept Pathol, New York, NY 10029 USA. Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. RP Fleischmajer, R (reprint author), Mt Sinai Med Ctr, Dept Dermatol, Box 1047, New York, NY 10029 USA. NR 52 TC 41 Z9 48 U1 0 U2 1 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD NOV PY 2000 VL 115 IS 5 BP 771 EP 777 DI 10.1046/j.1523-1747.2000.00138.x PG 7 WC Dermatology SC Dermatology GA 372DL UT WOS:000165219000001 PM 11069613 ER PT J AU Diamond, I Owolabi, T Marco, M Lam, C Glick, A AF Diamond, I Owolabi, T Marco, M Lam, C Glick, A TI Conditional gene expression in the epidermis of transgenic mice using the tetracycline-regulated transactivators tTA and rTA linked to the keratin 5 promoter SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE epidermis; hair follicle; tetracycline regulatory system; transgenic mice ID MOUSE SKIN; MAMMALIAN-CELLS; TUMOR PROMOTION; CARCINOGENESIS; SYSTEM; 12-O-TETRADECANOYLPHORBOL-13-ACETATE; DOXYCYCLINE; PAPILLOMAS; ONCOGENE AB To produce conditional expression of genes in the mouse epidermis we have generated transgenic mouse lines in which the tetracycline-regulated transcriptional transactivators, tTA and rTA, are linked to the bovine keratin 5 promoter. The transactivator lines were crossed with the tetOlacZ indicator line to test for transactivation in vivo. In the absence of doxycycline, the K5/tTA line induced beta -galactosidase enzyme activity in the epidermis at a level 500-fold higher than controls, and oral and topical doxycycline caused a dose- and time-dependent suppression of beta -galactosidase mRNA levels and enzyme activity. In the K5/rTA lines, doxycycline induced beta -galactosidase activity between 3- and 50-fold higher depending on the founder line, and this occurred within 24-48 h after dosing. Histochemical analysis of all lines localized beta -galactosidase expression to the basal layer of the epidermis and the outer root sheath of the hair follicle, as well as other keratin 5 positive tissues. In several K5/rTA lines, skin-specific transactivation was restricted to the hair follicle. Treatment of these double transgenic mice with 12-O-tetradecanoyl-phorbol-13-acetate caused rapid migration of beta -galactosidase marked cells from the hair follicle through the interfollicular epidermis, demonstrating the usefulness of this specific double transgenic for fate mapping cells in the epidermis. These results show that the tetracycline regulatory system produces effective conditional gene expression in the mouse epidermis, and suggest that it should be amenable to suppression and activation of foreign genes during development and specific pathologic conditions relevant to the epidermis. C1 NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bethesda, MD 20892 USA. RP Glick, A (reprint author), NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bldg 37, Bethesda, MD 20892 USA. NR 33 TC 130 Z9 132 U1 1 U2 2 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD NOV PY 2000 VL 115 IS 5 BP 788 EP 794 DI 10.1046/j.1523-1747.2000.00144.x PG 7 WC Dermatology SC Dermatology GA 372DL UT WOS:000165219000003 PM 11069615 ER PT J AU Fumelli, C Marconi, A Salvioli, S Straface, E Malorni, W Offidani, AM Pellicciari, R Schettini, G Giannetti, A Monti, D Franceschi, C Pincelli, C AF Fumelli, C Marconi, A Salvioli, S Straface, E Malorni, W Offidani, AM Pellicciari, R Schettini, G Giannetti, A Monti, D Franceschi, C Pincelli, C TI Carboxyfullerenes protect human keratinocytes from ultraviolet-B-induced apoptosis SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE apoptosis; carboxyfullerenes; keratinocytes; ultraviolet radiation ID DEOXY-D-RIBOSE; CULTURED HUMAN KERATINOCYTES; BLOOD MONONUCLEAR-CELLS; ALPHA-TOCOPHEROL; OXIDATIVE STRESS; N-ACETYLCYSTEINE; BCL-2 PROTEIN; SUNBURN CELL; SKIN; DEATH AB Carboxyfullerene, a water-soluble carboxylic acid derivative of a fullerene, which acts as a free-radical scavenger, was investigated as a protective agent against ultraviolet-light-induced damage in human keratinocytes. First, we demonstrate that carboxyfullerene is not cytotoxic for these cells. In addition, this compound significantly reduces the ultraviolet-B-induced inhibition of keratinocyte proliferation and protects keratinocytes from apoptosis caused by ultraviolet B irradiation in a time- and dose-dependent fashion. Furthermore, the percentage of cells with depolarized mitochondria is significantly lower in ultraviolet-B-irradiated keratinocytes pretreated with carboxyfullerene than in cells provided with diluent alone. Carboxyfullerene also protects human keratinocytes from apoptosis induced by exposure to deoxy-D-ribose, a sugar that causes cell death through a pathway involving oxidative stress. On the other hand, ultraviolet B downregulates bcl-2 levels in human keratinocytes, and carboxyfullerene fails to prevent this effect. These results suggest that carboxy- fullerene protects human keratinocytes from ultraviolet B damage possibly via a mechanism interfering with the generation of reactive oxygen species from depolarized mitochondria without the involvement of bcl-2. C1 Univ Modena & Reggio Emilia, Dept Neuropsychosensorial Pathol, Dermatol Sect, I-41100 Modena, Italy. Univ Modena & Reggio Emilia, Dept Biomed Sci, Sect Gen Pathol, I-41100 Modena, Italy. Ist Super Sanita, Dept Ultrastruct, I-00161 Rome, Italy. Univ Ancona, Dept Dermatol, Ancona, Italy. Univ Perugia, I-06100 Perugia, Italy. Univ Genoa, Dept Oncol, Natl Canc Inst, Unit Pharmacol & Neurosci, Genoa, Italy. Univ Florence, Dept Pathol & Expt Oncol, Florence, Italy. Univ Bologna, Dept Expt Pathol, I-40126 Bologna, Italy. RP Pincelli, C (reprint author), Univ Modena & Reggio Emilia, Dept Neuropsychosensorial Pathol, Dermatol Sect, Via Pozzo 71, I-41100 Modena, Italy. RI Marconi, Alessandra/A-5389-2012; Monti, Daniela/G-9556-2012; Malorni, Walter/G-5874-2016; Pincelli, Carlo/A-7288-2012; Salvioli, Stefano/K-6663-2016 OI Marconi, Alessandra/0000-0002-5667-5766; Monti, Daniela/0000-0001-8651-8123; Pincelli, Carlo/0000-0003-4416-2637; Salvioli, Stefano/0000-0002-6191-1225 NR 66 TC 48 Z9 53 U1 2 U2 6 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD NOV PY 2000 VL 115 IS 5 BP 835 EP 841 DI 10.1046/j.1523-1747.2000.00140.x PG 7 WC Dermatology SC Dermatology GA 372DL UT WOS:000165219000009 PM 11069621 ER PT J AU Ortaldo, JR Winkler-Pickett, R Wiegand, G AF Ortaldo, JR Winkler-Pickett, R Wiegand, G TI Activating Ly-49D NK receptors: expression and function in relation to ontogeny and Ly-49 inhibitor receptors SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE Ly-49; NK; ontogeny; product rule ID NATURAL-KILLER-CELLS; T-CELLS; TYROSINE PHOSPHATASE; MOUSE; CYTOTOXICITY; PHOSPHORYLATION; DIFFERENTIATION; RECRUITMENT; RECOGNITION; POPULATION AB Developmental changes in the repertoire of activating Ly-49 family members have not been examined previously. In the present study, we have examined the expression and function of the activating Ly-49s (D and H) from birth through 8 weeks of age, We demonstrate that 1) activating Ly-49s are expressed early, 2) their expression intensity is not different from adult NK cells, and 3) activating receptors are functional. Examination of the inhibitory Ly-49s also demonstrated functional capacity immediately upon expression. To examine the kinetics of expression of the repertoire of activating Ly-49 members, we utilized five- and six-color flow cytometric analyses of NK cells from birth through adulthood, Previous studies examining the inhibitory Ly-49 repertoire have proposed that expression is regulated by the product rule. Our results indicated that Ly-49D, which recognizes H-2D(d), had a discordantly high coexpression of the inhibitory Ly-49s that recognized H-2D(d) (Ly-49A and Ly-49G2). The product rule of Ly-49 expression does not explain the coexpression of selected activating and inhibitory receptors, This high level of coexpression of H-2D(d) recognizing activating and inhibitory Ly-49s suggests an in vivo selection or regulated coexpression. C1 NCI, Expt Immunol Lab, DBS, FCRDC, Frederick, MD 21702 USA. NCI, SAIC, FCRDC, Frederick, MD 21702 USA. RP Ortaldo, JR (reprint author), NCI, Expt Immunol Lab, DBS, FCRDC, Bldg 560,Rm 31-93, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 28 TC 16 Z9 16 U1 0 U2 0 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 NOV PY 2000 VL 68 IS 5 BP 748 EP 756 PG 9 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 370PQ UT WOS:000165131800022 PM 11073116 ER PT J AU Makrigiannis, AP Etzler, J Winkler-Pickett, R Mason, A Ortaldo, JR Anderson, SK AF Makrigiannis, AP Etzler, J Winkler-Pickett, R Mason, A Ortaldo, JR Anderson, SK TI Identification of the Ly49L protein: evidence for activating counterparts to inhibitory Ly49 proteins SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE NK cells; Ly49; cell surface receptors; signal transduction ID NATURAL-KILLER-CELLS; NK CELLS; TYROSINE PHOSPHORYLATION; SIGNAL-TRANSDUCTION; LY-49D RECEPTOR; LYMPHOCYTES-T; EXPRESSION; CLONING; DAP12; GENES AB Previous studies ha ve indicated that NK cells from different strains of inbred mice may express distinct Ly49 repertoires. Screening of Nh cells from the CBA/J mouse for inhibitory and activating Ly49s revealed a novel DAP12-associated receptor that was immunoprecipitated with the Ly49G-specific mAb 4D11. Degenerate primers were designed to amplify and clone Ly49 cDNAs from CBA/J NK cells. A novel activating Ly49 cDNA was identified, which bears strong homology to the partially sequenced Ly49l gene found in C57BL/6 mice, Transfection of Ly49l into a DAP12(+) cell. line and subsequent immunoprecipitation experiments showed that Ly49L is likely the activating Ly49 detected by the 4D11 antibody in CBA/J NK cells. Antibody-mediated cross-linking of Ly49L induced DAP12 phosphorylation, providing evidence that Ly49L is a functional activating receptor, Comparison of the extracellular domains of Ly49 family members indicates that all known activating members have an inhibitory counterpart with a highly related extracellular region. C1 NCI, Expt Immunol Lab, Div Basic Sci, SAIC Frederick,FCRDC, Frederick, MD 21702 USA. NCI, Intramural Res Support Program, SAIC Frederick, FCRDC, Frederick, MD 21702 USA. RP Anderson, SK (reprint author), NCI, Expt Immunol Lab, Div Basic Sci, SAIC Frederick,FCRDC, Bldg 560,Rm 31-93, Frederick, MD 21702 USA. RI Anderson, Stephen/B-1727-2012 OI Anderson, Stephen/0000-0002-7856-4266 FU NCI NIH HHS [N01CO56000] NR 32 TC 25 Z9 25 U1 0 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 NOV PY 2000 VL 68 IS 5 BP 765 EP 771 PG 7 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 370PQ UT WOS:000165131800024 PM 11073118 ER PT J AU Dagher, AP Aletras, A Choyke, P Balaban, RS AF Dagher, AP Aletras, A Choyke, P Balaban, RS TI Imaging of urea using chemical exchange-dependent saturation transfer at 1.5 T SO JOURNAL OF MAGNETIC RESONANCE IMAGING LA English DT Article DE kidney function; metabolism; tissue characterization; magnetization transfer; chemical exchange ID WATER AB The purpose of this study was to screen for slow proton chemical exchange between water and kidney metabolites using a standard clinical 1.5-T scanner. Imaging was performed using a fast spin-echo sequence with a magnetization transfer (MT) preparation pulse train. Off-resonance saturation ranging from +/- 50 to +/- 1000 Hz was used on urea and urine phantoms and normal human subjects imaged through the kidneys. The positive frequency was used as the control for each frequency pair. Results of frequency sweeps show an asymmetric NIT effect peaking at approximately 100 Hz (similar to1 ppm) for urea, urine, and renal parenchyma. Varying differences (50%-25%) occurred with different human subjects. Few differences were observed from phantom water or subject muscle tissue. Chemical exchange is detectable in the kidney near 1 ppm at 1.5 T, attributable to urea. This technique was used to produce in vivo distribution maps of this metabolite in vivo. (C) 2000 Wiley-Liss, Inc. C1 NIH, Dept Diagnost Radiol, Bethesda, MD 20892 USA. RP Dagher, AP (reprint author), NIH, Dept Diagnost Radiol, Bldg 10,Rm 1C640, Bethesda, MD 20892 USA. RI Balaban, Robert/A-7459-2009; OI Balaban, Robert/0000-0003-4086-0948; Aletras, Anthony/0000-0002-3786-3817 NR 4 TC 26 Z9 28 U1 1 U2 3 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1053-1807 J9 J MAGN RESON IMAGING JI J. Magn. Reson. Imaging PD NOV PY 2000 VL 12 IS 5 BP 745 EP 748 DI 10.1002/1522-2586(200011)12:5<745::AID-JMRI12>3.0.CO;2-H PG 4 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 477QH UT WOS:000171295400012 PM 11050645 ER PT J AU Marsiglio, W Amato, P Day, RD Lamb, ME AF Marsiglio, W Amato, P Day, RD Lamb, ME TI Scholarship on fatherhood in the 1990s and beyond SO JOURNAL OF MARRIAGE AND THE FAMILY LA English DT Review DE father-child relationship; fatherhood; father identify; fathering; father involvement ID CHILDRENS BEHAVIOR PROBLEMS; NONRESIDENT FATHERS; LIFE SATISFACTION; DOMESTIC VIOLENCE; DIVORCED FATHERS; INVOLVEMENT; FAMILY; MOTHERS; PARENT; PERSPECTIVE AB Throughout the 1990s, scholars interested in fatherhood have generated a voluminous, rich, and diverse body of work. We selectively review this literature with an eve toward prominent theoretical, methodological, and substantive issues, This burgeoning literature, complemented by social policy makers' heightened interest in fathers and families, focuses on fatherhood in at least 4 key ways. First, theorists have studied fatherhood as a cultural representation that is expressed through different sociocultural processes and embedded in a larger ecological context. Second, researchers have conceptualized and examined the diverse forms of fatherhood and father involvement. Third, attempts have been made to identify the linkages between dimensions of the father-child relationship and developmental outcomes among children and fathers. Fourth, scholars have explored the father identity as part of a reciprocal process negotiated by men, children, mothers, and other interested parties, Our review highlights research that examines the relationships between dimensions of the father-child relationship and children's well-being and development. We conclude by discussing promising avenues of scholarship for the next generation of research on fatherhood. C1 Univ Florida, Dept Sociol, Gainesville, FL 32611 USA. Penn State Univ, Dept Sociol, University Pk, PA 16803 USA. Brigham Young Univ, Dept Marriage Family & Human Dev, Provo, UT 84604 USA. NICHHD, Bethesda, MD 20892 USA. RP Marsiglio, W (reprint author), Univ Florida, Dept Sociol, Gainesville, FL 32611 USA. NR 175 TC 351 Z9 357 U1 4 U2 33 PU NATL COUNCIL FAMILY RELATIONS PI MINNEAPOLIS PA 3989 CENTRAL AVE NE #550, MINNEAPOLIS, MN 55421 USA SN 0022-2445 J9 J MARRIAGE FAM JI J. Marriage Fam. PD NOV PY 2000 VL 62 IS 4 BP 1173 EP 1191 DI 10.1111/j.1741-3737.2000.01173.x PG 19 WC Family Studies; Sociology SC Family Studies; Sociology GA 373DN UT WOS:000165273300018 ER PT J AU Roth, JS Wang, J Kelley, JA AF Roth, JS Wang, J Kelley, JA TI Determination of lodenosine and its major metabolite in human plasma by liquid chromatography/electrospray ionization tandem mass spectrometry SO JOURNAL OF MASS SPECTROMETRY LA English DT Article DE lodenosine; tandem mass spectrometry; plasma; electrospray ionization; pharmacokinetics ID 2'-BETA-FLUORO-2',3'-DIDEOXYADENOSINE; NUCLEOSIDE; HIV; PHARMACOKINETICS; SPECTRA; AGENTS; URINE; DRUG AB A sensitive and selective method for the determination of 2'-beta -fluoro-2',3'-dideoxyadenosine (lodenosine, F-ddA), an experimental anti-AIDS drug, and its major metabolite, 2'-beta -fluoro-2',3'-dideoxyinosine (F-ddI), in human plasma was developed and validated. The procedure employs two internal standards and a simple ultrafiltration step followed by chromatography on a Betasil C-18 minibore column. An in-line valve is used to remove salts before reaching the ion source. Detection is by electrospray ionization tandem mass spectrometry with selected reaction monitoring. The method has a limit of quantitation of 4 ng ml(-1) (16 nM) for F-ddA and 8 ng ml(-1) (32 nM) for F-ddI with a linear range up to 2000 ng ml(-1) (7.9 muM) for each. Predicted concentrations from a three-day validation study were within 5% of the nominal values for F-ddA and 16% for F-ddI, Intra- and interassay precision, as measured by relative standard deviation, was 13% or better for both compounds. To achieve good reproducibility, many variables related to the electrospray ionization were optimized for both precision and sensitivity. The method was successfully employed to analyze samples and evaluate plasma pharmacokinetics from a Phase I clinical trial. Copyright (C) 2000 John Wiley & Sons, Ltd. C1 NCI, Frederick Canc Res & Dev Ctr, Med Chem Lab, Div Basic Sci, Frederick, MD 21702 USA. RP Roth, JS (reprint author), NIH, Lab Neurotoxicol, MSC1262,10 Ctr Dr, Bethesda, MD 20892 USA. NR 23 TC 2 Z9 2 U1 0 U2 0 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 1076-5174 J9 J MASS SPECTROM JI J. Mass Spectrom. PD NOV PY 2000 VL 35 IS 11 BP 1313 EP 1319 DI 10.1002/1096-9888(200011)35:11<1313::AID-JMS65>3.0.CO;2-V PG 7 WC Biophysics; Chemistry, Organic; Spectroscopy SC Biophysics; Chemistry; Spectroscopy GA 380WF UT WOS:000165725900011 PM 11114090 ER PT J AU Sraiva, E Fampa, P Cedeno, V Bergoin, M Mialhe, E Miller, LH AF Sraiva, E Fampa, P Cedeno, V Bergoin, M Mialhe, E Miller, LH TI Expression of heterologous promoters in Lutzomyia longipalpis and Phlebotomus papatasi (Diptera : psychodidae) cell lines SO JOURNAL OF MEDICAL ENTOMOLOGY LA English DT Article DE Lutzomyia; Phlebotomus; sand fly cells; heterologous promoters; luciferase expression ID OYSTER CRASSOSTREA-GIGAS; YELLOW-FEVER MOSQUITO; TRANSGENIC PLANTS; AEDES-AEGYPTI; TRANSIENT EXPRESSION; GENETIC MANIPULATION; ANOPHELES-GAMBIAE; VECTOR; TRANSFORMATION; PROSPECTS AB To establish a transient expression system for genes introduced into sand fly cell lines, we tested the expression of the luciferase reporter gene under control of different promoters. Towards this end, we lipofected cell lines obtained from New and Old World sand flies, LL-5 from Lutzomyia longipalpis Lutz & Neiva and PP-9 from Phlebotomus papatasi Scopoli, respectively. The relative levels of luciferase expression were studied under control of Drosophila melanogaster Meigen heal shock protein 70 (hsp70), human cytomegalovirus, simian virus 40 or Junonia coenia (Hubner) densovirus (P9) promoters. The Drosophila heat shock protein 70 promoter, originating from insect genes, functioned as a strong promoter in both cell lines. Promoters From the different virus genes also were capable of driving transgene expression in both cell lines. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. Univ Fed Rio de Janeiro, Dept Imunol, BR-21949970 Rio De Janeiro, Brazil. INRA, UA CNRS 1184, Stn Rech Pathol Comparee, F-30380 St Christol Les Ales, France. Escuela Politecn Litoral, Cenaim, Guayaquil, Ecuador. RP Sraiva, E (reprint author), NIAID, Parasit Dis Lab, NIH, 9000 Rockville Pike,Bldg 4,Room 126, Bethesda, MD 20892 USA. RI Lima, Patricia/A-5165-2014 NR 27 TC 1 Z9 1 U1 0 U2 5 PU ENTOMOL SOC AMER PI LANHAM PA 9301 ANNAPOLIS RD, LANHAM, MD 20706 USA SN 0022-2585 J9 J MED ENTOMOL JI J. Med. Entomol. PD NOV PY 2000 VL 37 IS 6 BP 802 EP 806 PG 5 WC Entomology; Veterinary Sciences SC Entomology; Veterinary Sciences GA 380JT UT WOS:000165698600004 ER PT J AU Lafferty, AR Torpy, DJ Stowasser, M Taymans, SE Lin, JP Huggard, P Gordon, RD Stratakis, CA AF Lafferty, AR Torpy, DJ Stowasser, M Taymans, SE Lin, JP Huggard, P Gordon, RD Stratakis, CA TI A novel genetic locus for low renin hypertension: familial hyperaldosteronism type II maps to chromosome 7 (7p22) SO JOURNAL OF MEDICAL GENETICS LA English DT Article DE chromosome 7; aldosterone; familial hyperaldosteronism type II; hypertension ID PRIMARY ALDOSTERONISM; STEROID 11-BETA-HYDROXYLASE; LINKAGE ANALYSIS; PHYSICAL MAP; CYP11B2 GENE; HUMAN GENOME; SYNTHASE; BIOSYNTHESIS; MUTATIONS; P450C11AS AB Familial hyperaldosteronism type II (FH-II) is caused by adrenocortical hyperplasia or aldosteronoma or both and is frequently transmitted in an autosomal dominant fashion. Unlike FH type I (FI-I-I), which results from fusion of the CYP11B1 and CYP11B2 genes, hyperaldosteronism in FH-II is not glucocorticoid remediable. A large family with FH-II was used for a genome wide search and its members were evaluated by measuring the aldosterone:renin ratio. In those with an increased ratio, FH-II was confirmed by fludrocortisone suppression testing. After excluding most of the genome, genetic linkage was identified with a maximum two point lod score of 3.26 at theta =0, between FH-II in this family and the polymorphic markers D7S511, D7S517, and GATA24F03 on chromosome 7,a region that corresponds to cytogenetic band 7p22. This is the first identified locus for FH-II; its molecular elucidation may provide further insight into the aetiology of primary aldosteronism. C1 NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, Bethesda, MD 20892 USA. Univ Queensland, Greenslopes Hosp, Dept Med, Brisbane, Qld 4120, Australia. NHLBI, Off Biostat Res, Div Epidemiol & Clin Applicat, NIH, Bethesda, MD 20892 USA. RP Stratakis, CA (reprint author), NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, Bldg 10,Room 10N262,10 Ctr Dr,MSC1862, Bethesda, MD 20892 USA. RI Stowasser, Michael/F-4121-2010; Gordon, Richard/K-2555-2012 NR 33 TC 94 Z9 99 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0022-2593 J9 J MED GENET JI J. Med. Genet. PD NOV PY 2000 VL 37 IS 11 BP 831 EP 835 DI 10.1136/jmg.37.11.831 PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 374LD UT WOS:000165345900003 PM 11073536 ER PT J AU Park, KY Dalakas, MC Goebel, HH Ferrans, VJ Semino-Mora, C Litvak, S Takeda, K Goldfarb, LG AF Park, KY Dalakas, MC Goebel, HH Ferrans, VJ Semino-Mora, C Litvak, S Takeda, K Goldfarb, LG TI Desmin splice variants causing cardiac and skeletal myopathy SO JOURNAL OF MEDICAL GENETICS LA English DT Article DE cardiac and skeletal myopathy; desmin splice site mutations; expression study; genotype-phenotype correlation ID FAMILIAL CARDIOMYOPATHY; INTERMEDIATE FILAMENTS; MYOFIBRILLAR MYOPATHY; GENE; MUTATION; SEQUENCE; CELLS; CONSEQUENCES; POSITIVITY; EXPRESSION AB Desmin myopathy is a hereditary or sporadic cardiac and skeletal myopathy characterised by intracytoplasmic accumulation of desmin reactive deposits in muscle cells. We have characterised novel splice site mutations in the gene desmin resulting in deletion of the entire exon 3 during the pre-mRNA splicing. Sequencing of cDNA and genomic DNA identified a heterozygous de novo A to G change at the +3 position of the splice donor site of intron 3 (IVS3+3A-->G) in a patient with sporadic skeletal and cardiac myopathy. A G to A transition at the highly conserved -1 nucleotide position of intron 2 affecting the splice acceptor site (IVS2-1G-->A) was found in an unrelated patient with a similar phenotype. Expression of genomic DNA fragments carrying the IVS3+3A-->G and IVS2-1G-->A mutations confirmed that these mutations cause exon 3 deletion. Aberrant splicing leads to an in frame deletion of 32 complete codons and is predicted to result in mutant desmin lacking 32 amino acids from the 1B segment of the alpha helical rod. Functional analysis of the mutant desmin in SW13 (vim-) cells showed aggregation of abnormal coarse clumps of desmin positive material dispersed throughout the cytoplasm. This is the first report on the pathogenic potentials of splice site mutations in the desmin gene. C1 NINDS, Clin Neurogenet Unit, NIH, Bethesda, MD 20892 USA. NINDS, Neuromuscular Disorders Sect, NIH, Bethesda, MD 20892 USA. Univ Mainz, Med Ctr, Dept Neuropathol, D-55131 Mainz, Germany. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Goldfarb, LG (reprint author), NINDS, Clin Neurogenet Unit, NIH, Bldg 10,Room 4B37,10 Ctr Dr,MSC 1361, Bethesda, MD 20892 USA. NR 29 TC 48 Z9 52 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0022-2593 J9 J MED GENET JI J. Med. Genet. PD NOV PY 2000 VL 37 IS 11 BP 851 EP 857 DI 10.1136/jmg.37.11.851 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 374LD UT WOS:000165345900006 PM 11073539 ER PT J AU She, MD Trimble, D Yu, LC Chalovich, JM AF She, MD Trimble, D Yu, LC Chalovich, JM TI Factors contributing to troponin exchange in myofibrils and in solution SO JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY LA English DT Article ID RABBIT PSOAS MUSCLE; THIN FILAMENT; SKELETAL-MUSCLE; MYOSIN SUBFRAGMENT-1; CARDIAC TROPONIN; SKINNED FIBERS; CONTRACTION; TROPOMYOSIN; COMPLEX; ACTIN AB The troponin complex in a muscle fiber can be replaced with exogenous troponin by using a gentle exchange procedure in which the actin-tropomyosin complex is never devoid of a full complement of troponin (Brenner et al. (1999) Biophys J 77: 2677-2691). The mechanism of this exchange process and the factors that influence this exchange are poorly understood. In this study, the exchange process has now been examined in myofibrils and in solution. In myofibrils under rigor conditions, troponin exchange occurred preferentially in the region of overlap between actin and myosin when the free Ca2+ concentration was low. At higher concentrations of Ca2+, the exchange occurred uniformly along the actin. Ca2+ also accelerated troponin exchange in solution but the effect of S1 could not be confirmed in solution experiments. The rate of exchange in solution was insensitive to moderate changes in pH or ionic strength. Increasing the temperature resulted in a two-fold increase in rate with each 10 degreesC increase in temperature. A sequential two step model of troponin binding to actin-tropomyosin could simulate the observed association and dissociation transients. In the absence of Ca2+ or rigor S1, the following rate constants could describe the binding process: k(1) = 7.12 muM(-1)s(-1), k(-1) = 0.65 s(-1), k(2) = 0.07 s(-1), k(-2) = 0.0014 s(-1). The slow rate of detachment of troponin from actin (k(-2)) limits the rate of exchange in solution and most likely contributes to the slow rate of exchange in fibers. C1 NIAMSD, Phys Biol Lab, NIH, Bethesda, MD 20892 USA. E Carolina Univ, Sch Med, Dept Biochem, Greenville, NC 27834 USA. RP Yu, LC (reprint author), NIAMSD, Phys Biol Lab, NIH, Bethesda, MD 20892 USA. OI Chalovich, Joseph/0000-0002-1243-4055 FU NIAMS NIH HHS [AR 44 504] NR 29 TC 18 Z9 18 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0142-4319 J9 J MUSCLE RES CELL M JI J. Muscle Res. Cell Motil. PD NOV PY 2000 VL 21 IS 8 BP 737 EP 745 DI 10.1023/A:1010300802980 PG 9 WC Cell Biology SC Cell Biology GA 419DV UT WOS:000167934800004 PM 11392555 ER PT J AU Groweiss, A Cardellina, JH Boyd, MR AF Groweiss, A Cardellina, JH Boyd, MR TI HIV-inhibitory prenylated xanthones and flavones from Maclura tinctoria SO JOURNAL OF NATURAL PRODUCTS LA English DT Article ID NATURAL-PRODUCTS; CUDRANIA-TRICUSPIDATA; ROOT BARK; MORACEAE PLANTS; COMPONENTS; PRENYLFLAVONES; BENZOPHENONES; CONSTITUENTS AB The organic extract of the plant Maclura tinctoria exhibited moderate anti-HIV activity. Seven prenylated phenolic derivatives were isolated from the active fractions and characterized by spectroanalytical methods. New compounds macluraxanthone B (1), macluraxanthone C (2), and dihydrocudraflavone B (8) were identified. C1 NCI, Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag, Lab Drug Discovery Res & Dev,Dev Therapeut Progra, Frederick, MD 21702 USA. RP Boyd, MR (reprint author), NCI, Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag, Lab Drug Discovery Res & Dev,Dev Therapeut Progra, Frederick, MD 21702 USA. NR 19 TC 92 Z9 98 U1 1 U2 14 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 NOV PY 2000 VL 63 IS 11 BP 1537 EP 1539 DI 10.1021/np000175m PG 3 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 377VY UT WOS:000165548000020 PM 11087602 ER PT J AU Rall, JE AF Rall, JE TI Mystery of mysteries: Is evolution a social construction? SO JOURNAL OF NERVOUS AND MENTAL DISEASE LA English DT Book Review C1 NIH, Bethesda, MD 20892 USA. RP Rall, JE (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3018 J9 J NERV MENT DIS JI J. Nerv. Ment. Dis. PD NOV PY 2000 VL 188 IS 11 BP 788 EP 789 DI 10.1097/00005053-200011000-00011 PG 2 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 372VN UT WOS:000165254900011 ER PT J AU Schomerus, C Korf, HW Laedtke, E Weller, JL Klein, DC AF Schomerus, C Korf, HW Laedtke, E Weller, JL Klein, DC TI Selective adrenergic/cyclic AMP-dependent switch-off of proteasomal proteolysis alone switches on neural signal transduction: An example from the pineal gland SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE arylalkylamine N-acetyltransferase; biological rhythms; neural signaling; noradrenaline; pineal gland; proteasomal proteolysis ID SEROTONIN N-ACETYLTRANSFERASE; MELATONIN SYNTHESIS; MESSENGER-RNA; RAT PINEALOCYTES; CEREBROSPINAL-FLUID; NOCTURNAL INCREASE; RHYTHM; PHOSPHORYLATION; EXPRESSION; UBIQUITIN AB The molecular processes underlying neural transmission are central issues in neurobiology. Here we describe a novel mechanism through which noradrenaline (NA) activates its target cells, using the mammalian pineal organ as a model. In this neuroendocrine transducer, NA stimulates arylalkylamine N-acetyltransferase (AANAT; EC 2.3.1.87), the key enzyme regulating the nocturnal melatonin production. In rodents, AANAT protein accumulates as a result of enhanced transcription, but in primates and ungulates, the AANAT mRNA level fluctuates only marginally, indicating that other mechanisms regulate AANAT protein and activity. These were investigated in cultured bovine pinealocytes. AANAT mRNA was readily detectable in unstimulated pinealocytes, and levels did not change following NA treatment. In contrast, NA increased AANAT protein levels in parallel with AANAT activity, apparently through a cyclic AMP-mediated mechanism. Immunocytochemistry revealed that the changes in AANAT protein levels occurred in virtually all pinealocytes. Inhibition of AANAT degradation by proteasomal proteolysis alone was found to switch-on enzyme activity by increasing AANAT protein levels five- to 10-fold. Accordingly, under unstimulated conditions AANAT protein is continually synthesized and immediately destroyed by proteasomal proteolysis. NA appears to act via cyclic AMP to protect AANAT from proteolytic destruction, resulting in accumulation of the protein. These findings show that tightly regulated control of proteasomal proteolysis of a specific protein alone can play a pivotal role in neural regulation. C1 Univ Frankfurt, Inst Anat 2, D-60590 Frankfurt, Germany. NICHHD, Sect Neuroendocrinol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Korf, HW (reprint author), Univ Frankfurt, Inst Anat 2, Theodor Stern Kai 7, D-60590 Frankfurt, Germany. FU NIMH NIH HHS [N0-IMH30003] NR 46 TC 65 Z9 68 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD NOV PY 2000 VL 75 IS 5 BP 2123 EP 2132 DI 10.1046/j.1471-4159.2000.0752123.x PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 365AA UT WOS:000089925200037 PM 11032902 ER PT J AU Gallant, PE AF Gallant, PE TI Axonal protein synthesis and transport SO JOURNAL OF NEUROCYTOLOGY LA English DT Article ID SQUID GIANT-AXON AB Recent evidence has challenged our ideas about the nature of axonal protein synthesis and transport. Previous metabolic labeling evidence supported the idea that all axonal proteins were synthesized in the cell body and then transported as formed cytoplasmic structures into the axon. Recent evidence suggests that neither the synthesis nor the transport of axonal proteins is that simple. Though most axonal proteins do appear to be synthesized in the neuronal cell body, a small amount of protein appears to be synthesized intra-axonally in some axons. Though small in amount, intra-axonal protein synthesis may be important functionally in some axons. Recent experiments have also begun to identify the presence of a rich array of transport motors in axons, including many members of the kinesin, dynein and myosin families. Progress is being made in identifying which cargoes are being transported by which of these motors. Finally, recent experiments have addressed an old question about whether axoplasmic proteins are transported as filamentous polymers or as soluble components in axons. The answer is that both mechanism can be used in axons. For example, neurofilament protein can move in its particulate or polymeric state, while tubulin can move in its soluble or unpolymerized state. C1 NINDS, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Gallant, PE (reprint author), NINDS, Neurobiol Lab, NIH, Bldg 36 Rm 2A-21, Bethesda, MD 20892 USA. NR 12 TC 27 Z9 28 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0300-4864 J9 J NEUROCYTOL JI J. Neurocytol. PD NOV PY 2000 VL 29 IS 11-12 BP 779 EP 782 DI 10.1023/A:1010961022455 PG 4 WC Cell Biology; Neurosciences SC Cell Biology; Neurosciences & Neurology GA 452NG UT WOS:000169864000001 PM 11466469 ER PT J AU Grant, P Pant, HC AF Grant, P Pant, HC TI Neurofilament protein synthesis and phosphorylation SO JOURNAL OF NEUROCYTOLOGY LA English DT Review ID CYCLIN-DEPENDENT KINASE-5; NERVE GROWTH-FACTOR; ROOT GANGLION NEURONS; SQUID GIANT-AXON; MICROTUBULE-ASSOCIATED PROTEINS; INTERMEDIATE FILAMENT PROTEIN; SUBUNIT NF-H; MONOCLONAL-ANTIBODIES DISTINGUISH; SITE-SPECIFIC PHOSPHORYLATION; CULTURED HIPPOCAMPAL-NEURONS AB Neurofilament proteins, a major intermediate filament component of the neuronal cytoskeleton, are organized as 10 nm thick filaments in axons and dendrites. They are large, abundantly phosphorylated proteins with numerous phosphate acceptor sites, up to 100 in some cases, organized as numerous repeat motifs. Together with other cytoskeletal components such as microtubules, MAPs, actin and plectin-like linking molecules, they make up a dynamic lattice that sustains neuronal function from neuronal "birthday" to apoptotic cell death. The activity of the neuronal cytoskeleton is regulated by phosphorylation, dephosphorylation reactions mediated by numerous associated kinases, phosphatases and their regulators. Factors regulating multisite phosphorylation of NFs are topographically localized, with maximum phosphorylation of NF proteins consigned to axons. Phosphorylation defines the nature of NF interactions with one another and with other cytoskeletal components such as microtubules, MAPs and actin. To understand how these functional interactions are regulated by phosphorylation we attempt to identify the relevant kinases and phosphatases, their specific targets and the factors modulating their activity. As an initial working model we propose that NF phosphorylation is regulated topographically in neurons by compartment-specific macromolecular complexes of substrates, kinases and phosphatases. This implies that axonal complexes differ structurally and functionally from those in cell bodies and dendrites. Such protein assemblies, by virtue of conformational changes within proteins, facilitate ordered, sequential multisite phosphorylations that modulate dynamic cytoskeletal interactions. C1 NINDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Pant, HC (reprint author), NINDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. NR 266 TC 117 Z9 118 U1 2 U2 10 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0300-4864 J9 J NEUROCYTOL JI J. Neurocytol. PD NOV PY 2000 VL 29 IS 11-12 BP 843 EP 872 DI 10.1023/A:1010999509251 PG 30 WC Cell Biology; Neurosciences SC Cell Biology; Neurosciences & Neurology GA 452NG UT WOS:000169864000007 PM 11466475 ER PT J AU Galbraith, JA Gallant, PE AF Galbraith, JA Gallant, PE TI Axonal transport of tubulin and actin SO JOURNAL OF NEUROCYTOLOGY LA English DT Review ID EXPERIMENTAL DIABETIC NEUROPATHY; RAT SCIATIC-NERVE; CYTOSKELETAL PROTEIN-TRANSPORT; RETINAL GANGLION-CELLS; MOUSE OPTIC-NERVE; SLOW COMPONENT-B; SQUID GIANT-AXON; GROWING AXONS; FLUORESCENT MICROTUBULES; NEURONAL CENTROSOME AB Axonal transport is responsible for supplying the axonal processes with proteins that are synthesized in the cell body. Among the proteins that are moved by this mechanism are tubulin and actin, two major components of the cytoskeleton. Observation of the movement of metabolically labeled tubulin and actin in-vivo has demonstrated that tubulin and actin transport are reduced in various diseases and with age, but transport is increased during axonal growth and regeneration. These metabolic studies have also raised questions about the underlying mechanisms of slow axonal transport such as: what is the polymerization state of tubulin and actin during transport, what motors and tracks are responsible for their movement down the axon, and how are the transport motors coupled to tubulin and actin during transport ? Since experiments using metabolically labeled tubulin and actin have not effectively addressed these questions, a variety of new in-vitro fluorescent microscopy techniques have been devised to investigate these questions. These fluorescent microscopy experiments have suggested that tubulin can be transported in the unpolymerized soluble state and that such transport of soluble tubulin relies on the presence of formed microtubule tracks. It is not yet known what motor or motors are responsible for tubulin or actin transport in axons or how such a motor(s) might be coupled to such an abundant soluble cargo. C1 NINDS, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Galbraith, JA (reprint author), NINDS, Neurobiol Lab, NIH, Bldg 36 Rm 2A-18, Bethesda, MD 20892 USA. NR 122 TC 25 Z9 27 U1 2 U2 4 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0300-4864 J9 J NEUROCYTOL JI J. Neurocytol. PD NOV PY 2000 VL 29 IS 11-12 BP 889 EP 911 DI 10.1023/A:1010903710160 PG 23 WC Cell Biology; Neurosciences SC Cell Biology; Neurosciences & Neurology GA 452NG UT WOS:000169864000009 PM 11466477 ER PT J AU Le, YY Hu, JY Gong, WH Shen, WP Li, BQ Dunlop, NM Halverson, DO Blair, DG Wang, JM AF Le, YY Hu, JY Gong, WH Shen, WP Li, BQ Dunlop, NM Halverson, DO Blair, DG Wang, JM TI Expression of functional formyl peptide receptors by human astrocytoma cell lines SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE astrocytoma cells; chemotaxis; calcium mobilization; formyl peptide receptors; cytokines ID PROTEIN-COUPLED RECEPTOR; OXIDE-MEDIATED MECHANISM; LIPOXIN A(4) RECEPTOR; TUMOR-NECROSIS-FACTOR; AMYLOID-BETA-PEPTIDE; FORMYLPEPTIDE RECEPTOR; 7-TRANSMEMBRANE; MONOCYTES; CYTOKINES; DISEASE AB Activation of astrocytes is important in the pathogenesis of a variety of diseases in the central nervous system, such as infection and neurodegeneration. We found that the bacterial chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (fMLF) induced potent migration and Ca2+ mobilization in human astrocytoma cell lines. The effect of fMLF was pertussis toxin-sensitive, suggesting the involvement of seven transmembrane, G protein-coupled receptor(s) for fMLF. Scatchard analyses revealed that astrocytoma cell Lines express both high- and low-affinity binding sites for [H-3]fMLF. RT-PCR confirmed the expression of transcripts of fMLF receptors, the high-affinity FPR and the low-affinity FPRL1 by these cells. Both fMLF and F peptide, a synthetic peptide domain of HIV-1 envelope protein which specifically activates FPRL1, increased secretion of IL-6 by astrocytoma cells. Our study demonstrates for the first time that FPR and FPRL1 expressed by astrocytoma cell lines are functional, and suggests a molecular basis for the involvement of these receptors in host defense in the brain. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Basic Res Lab, Frederick, MD 21702 USA. RP Wang, JM (reprint author), NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Mol Immunoregulat Lab, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 33 TC 68 Z9 71 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-5728 J9 J NEUROIMMUNOL JI J. Neuroimmunol. PD NOV 1 PY 2000 VL 111 IS 1-2 BP 102 EP 108 DI 10.1016/S0165-5728(00)00373-8 PG 7 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 373YU UT WOS:000165318300013 PM 11063827 ER PT J AU Muraro, PA Leist, T Bielekova, B McFarland, HF AF Muraro, PA Leist, T Bielekova, B McFarland, HF TI VLA-4/CD49d downregulated on primed T lymphocytes during interferon-beta therapy in multiple sclerosis SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE T lymphocytes; multiple sclerosis; interferon-beta; adhesion molecules; VLA-4 ID DIFFERENTIAL EXPRESSION; CELL-ADHESION; MEMORY; MECHANISMS; INTEGRINS; SUBSETS; VLA-4 AB Effects on adhesion molecules of immune cells might contribute to the mode of action of interferon-beta (IFN-beta) in multiple sclerosis (MS). We have serially monitored the cell surface expression of integrins CD49d (VLA-4) and CD11a (LFA-I) on fresh T lymphocyte subpopulations from 5 MS patients monthly for 2 months prior to treatment and for 3 months on treatment with IFN-beta 1b. In parallel, we assessed inflammatory disease activity by monthly contrast-enhanced magnetic resonance imaging (MRI). IFN-beta treatment specifically downregulated CD49d expression on CD8+ and CD4+/CD45RO+ 'memory' T lymphocytes and differentially modulated the proportion of CD4+, CD8+ and CD27+ T cells. These effects may play an important role in the reduction of central nervous system cell trafficking and inflammation in MS. (C) 2000 Elsevier Science BN. All rights reserved. C1 NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Univ G DAnnunzio, Sch Med, Dept Oncol & Neurosci, I-66013 Chieti, Italy. RP Muraro, PA (reprint author), NINDS, Neuroimmunol Branch, NIH, 10 Ctr Dr MSC1400, Bethesda, MD 20892 USA. OI Muraro, Paolo/0000-0002-3822-1218 NR 25 TC 46 Z9 46 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-5728 J9 J NEUROIMMUNOL JI J. Neuroimmunol. PD NOV 1 PY 2000 VL 111 IS 1-2 BP 186 EP 194 DI 10.1016/S0165-5728(00)00362-3 PG 9 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 373YU UT WOS:000165318300023 PM 11063837 ER PT J AU Wenner, P O'Donovan, MJ Matise, MP AF Wenner, P O'Donovan, MJ Matise, MP TI Topographical and physiological characterization of interneurons that express engrailed-1 in the embryonic chick spinal cord SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID RECURRENT INHIBITION; NERVOUS-SYSTEM; MOTOR-ACTIVITY; MOTONEURONS; GENE; NEURONS; SPECIFICATION; POPULATION; MOUSE; TRANSMISSION AB A number of homeodomain transcription factors have been implicated in controlling the differentiation of various types of neurons including spinal motoneurons. Some of these proteins are also expressed in spinal interneurons, but their function is unknown. Progress in understanding the role of transcription factors in interneuronal development has been slow because the synaptic connections of interneurons, which in part define their identity, are difficult to establish. Using whole cell recording in the isolated spinal cord of chick embryos, we assessed the synaptic connections of lumbosacral interneurons expressing the Engrailed-1 (En1) transcription factor. Specifically we established whether En1-expressing interneurons made direct connections with motoneurons and whether they constitute a single interneuron class. Cells were labeled with biocytin and subsequently processed for En1 immunoreactivity. Our findings indicate that the connections of En1-expressing cells with motoneurons and with sensory afferents were diverse, suggesting that the population was heterogeneous. In addition, the synaptic connections we tested were similar in interneurons that expressed the En1 protein and in many that did not. The majority of sampled En1 cells did, however, exhibit a direct synaptic connection to motoneurons that is likely to be GABAergic. Because our physiological methods underestimate the number of direct connections with motoneurons, it is possible that the great majority, perhaps all, En1-expressing cells make direct synaptic connections with motoneurons. Our results raise the possibility that En1 could be involved in interneuron-motoneuron connectivity but that its expression is not restricted to a distinct functional subclass of ventral interneuron. These findings constrain hypotheses about the role of En-1 in interneuron development and function. C1 Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Neurosci & Cell Biol, Piscataway, NJ 08854 USA. NINDS, NIH, Neural Control Lab, Bethesda, MD 20892 USA. RP Wenner, P (reprint author), Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Neurosci & Cell Biol, 675 Hoes Lane, Piscataway, NJ 08854 USA. RI o'donovan, michael/A-2357-2015; OI o'donovan, michael/0000-0003-2487-7547; Wenner, Peter/0000-0002-7072-2194 NR 31 TC 52 Z9 54 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD NOV PY 2000 VL 84 IS 5 BP 2651 EP 2657 PG 7 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 371CW UT WOS:000165161200043 PM 11068006 ER PT J AU Li, Q Wichems, C Heils, A Lesch, KP Murphy, DL AF Li, Q Wichems, C Heils, A Lesch, KP Murphy, DL TI Reduction in the density and expression, but not G-protein coupling, of serotonin receptors (5-HT1A) in 5-HT transporter knock-out mice: Gender and brain region differences SO JOURNAL OF NEUROSCIENCE LA English DT Article DE 5-HT1A receptors; 5-HT1A mRNA; 5-HT transporter knock-out mice; G-protein coupling; hypothermia; gender difference; autoradiography; competitive RT-PCR; in situ hybridization ID MESSENGER-RNA EXPRESSION; ANXIETY-RELATED TRAITS; NEUROENDOCRINE RESPONSES; RAT-BRAIN; REPEATED INJECTIONS; RHESUS MACAQUES; G(O) PROTEINS; FLUOXETINE; DESENSITIZATION; BINDING AB The aim of the present study was to investigate the mechanisms underlying the desensitization of 5-HT1A receptors in the dorsal raphe and hypothalamus of serotonin (5-HT) transporter knockout mice (5-HTT -/-). The density of 5-HT1A receptors in the dorsal raphe was reduced in both male and female 5-HTT -/- mice. This reduction was more extensive in female than in male 5-HTT -/- mice. 8-OH-DPAT-induced hypothermia was absent in female 5-HTT -/- and markedly attenuated in 5-HTT -/- mice. The density of 5-HT1A receptors also was decreased significantly in several nuclei of the hypothalamus, amygdala, and septum of female 5-HTT -/- mice. 5- HT1A receptor mRNA was reduced significantly in the dorsal raphe region, but not in the hypothalamus or hippocampus, of female 5-HTT +/- and 5- HTT -/- mice. G-protein coupling to 5- HT1A receptors and G-protein levels in most brain regions were not reduced significantly, except that G(o) and G(i1) proteins were reduced modestly in the midbrain of 5-HTT -/- mice. These data suggest that the desensitization of 5-HT1A receptors in 5-HTT -/- mice may be attributable to a reduction in the density of 5-HT1A receptors. This reduction is brain region-specific and more extensive in the female mice. The reduction in the density of 5-HT1A receptors may be mediated partly by reduction in the gene expression of 5-HT1A receptors in the dorsal raphe, but also by other mechanisms in the hypothalamus of 5-HTT -/- female mice. Finally, alterations in G-protein coupling to 5-HT1A receptors are unlikely to be involved in the desensitization of 5-HT1A receptors in 5-HTT -/- mice. C1 NIMH, Clin Sci Lab, NIH, Ctr Clin, Bethesda, MD 20892 USA. Univ Wurzburg, Dept Psychiat, D-97080 Wurzburg, Germany. RP Li, Q (reprint author), NIMH, Clin Sci Lab, NIH, Ctr Clin, Room 3D41,10 Ctr Dr,MSC 1264, Bethesda, MD 20892 USA. RI Lesch, Klaus-Peter/J-4906-2013 OI Lesch, Klaus-Peter/0000-0001-8348-153X NR 41 TC 156 Z9 161 U1 0 U2 3 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 NOV 1 PY 2000 VL 20 IS 21 BP 7888 EP 7895 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 366KB UT WOS:000090003600006 PM 11050108 ER PT J AU Blondel, O Collin, C McCarran, WJ Zhu, SP Zamostiano, R Gozes, I Brenneman, DE McKay, RDG AF Blondel, O Collin, C McCarran, WJ Zhu, SP Zamostiano, R Gozes, I Brenneman, DE McKay, RDG TI A glia-derived signal regulating neuronal differentiation SO JOURNAL OF NEUROSCIENCE LA English DT Article DE glia; synapse; hippocampus; ADNF; NT-3; VIP ID VASOACTIVE-INTESTINAL-PEPTIDE; CULTURED HIPPOCAMPAL-NEURONS; MINIATURE SYNAPTIC CURRENTS; LONG-TERM POTENTIATION; DEPENDENT NEUROTROPHIC FACTOR; NEUROPROTECTIVE PEPTIDE; NMDA RECEPTORS; IN-VITRO; SYNAPSES; MICE AB Astrocytes are present in large numbers in the nervous system, are associated with synapses, and propagate ionic signals. Astrocytes influence neuronal physiology by responding to and releasing neurotransmitters, but the mechanisms that establish the close interaction between these cells are not defined. Here we use hippocampal neurons in culture to demonstrate that vasoactive intestinal polypeptide (VIP) promotes neuronal differentiation through activity-dependent neurotrophic factor (ADNF), a protein secreted by VIP-stimulated astroglia. ADNF is produced by glial cells and acts directly on neurons to promote glutamate responses and morphological development. ADNF causes secretion of neurotrophin 3 (NT-3), and both proteins regulate NMDA receptor subunit 2A (NR2A) and NR2B. These data suggest that the VIP-ADNF-NT-3 neuronal-glial pathway regulates glutamate responses from an early stage in the synaptic development of excitatory neurons and may also contribute to the known effects of VIP on learning and behavior in the adult nervous system. C1 NINDS, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NICHHD, Sect Dev & Mol Pharmacol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP McKay, RDG (reprint author), NINDS, Mol Biol Lab, NIH, Bldg 36,Room 3D02, Bethesda, MD 20892 USA. NR 49 TC 156 Z9 163 U1 0 U2 5 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 NOV 1 PY 2000 VL 20 IS 21 BP 8012 EP 8020 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 366KB UT WOS:000090003600020 PM 11050122 ER PT J AU Ratliff, JK Oldfield, EH AF Ratliff, JK Oldfield, EH TI Multiple pituitary adenomas in Cushing's disease SO JOURNAL OF NEUROSURGERY LA English DT Article DE pituitary; adenoma; microadenoma; transsphenoidal surgery; Cushing's disease ID INTRAOPERATIVE ULTRASOUND; DIFFERENTIAL-DIAGNOSIS; PROLACTIN; HYPERPROLACTINEMIA; MICROADENOMAS; CORTICOTROPIN; MANAGEMENT; AUTOPSY; HORMONE; SURGERY AB Object. Clinically evident multiple pituitary adenomas rarely occur. The authors assess the incidence and clinical relevance of multiple adenomas in Cushing's disease. Methods. A prospective clinical database of 660 pituitary surgeries was analyzed to assess the incidence of multiple pituitary adenomas in Cushing's disease. Relevant radiographic scans, medical records, and histopathological reports were reviewed. Thirteen patients with at least two separate histopathologically confirmed pituitary adenomas were identified. Prolactinomas (nine patients) were the most common incidental tumors. Other incidental tumors included secretors of growth hormone ([GH], one patient) and GH and prolactin (two patients), and a null-cell tumor (one patient). In two patients, early repeated surgery was performed because the initial operation failed to correct hypercortisolism, in one instance because the tumor excised at the initial surgery was a prolactinoma, not an adrenocorticotropic hormone-secreting tumor. One patient had three distinct tumors. Conclusions. Multiple pituitary adenomas are rare, but may complicate management of patients with pituitary disease. C1 NINCDS, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Oldfield, EH (reprint author), NINCDS, Surg Neurol Branch, NIH, Bldg 10,Room 5D37, Bethesda, MD 20892 USA. NR 38 TC 34 Z9 35 U1 0 U2 1 PU AMER ASSOC NEUROLOGICAL SURGEONS PI CHARLOTTESVILLE PA UNIV VIRGINIA, 1224 WEST MAIN ST, STE 450, CHARLOTTESVILLE, VA 22903 USA SN 0022-3085 J9 J NEUROSURG JI J. Neurosurg. PD NOV PY 2000 VL 93 IS 5 BP 753 EP 761 DI 10.3171/jns.2000.93.5.0753 PG 9 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 366YL UT WOS:000090033800004 PM 11059654 ER PT J AU Oldfield, EH AF Oldfield, EH TI Multiple pituitary adenomas - Response SO JOURNAL OF NEUROSURGERY LA English DT Letter ID CUSHINGS-DISEASE; INTRAOPERATIVE ULTRASOUND C1 NIH, Bethesda, MD 20892 USA. RP Oldfield, EH (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 9 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC NEUROLOGICAL SURGEONS PI CHARLOTTESVILLE PA UNIV VIRGINIA, 1224 WEST MAIN ST, STE 450, CHARLOTTESVILLE, VA 22903 USA SN 0022-3085 J9 J NEUROSURG JI J. Neurosurg. PD NOV PY 2000 VL 93 IS 5 BP 910 EP 911 PG 2 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 366YL UT WOS:000090033800035 ER PT J AU Lodge, MA Carson, RE Carrasquillo, JA Whatley, M Libutti, SK Bacharach, SL AF Lodge, MA Carson, RE Carrasquillo, JA Whatley, M Libutti, SK Bacharach, SL TI Parametric images of blood flow in oncology PET studies using [O-15]water SO JOURNAL OF NUCLEAR MEDICINE LA English DT Article DE PET; [O-15]water; parametric image; tumor blood flow ID POSITRON-EMISSION TOMOGRAPHY; QUANTITATIVE MEASUREMENT; COMPUTED-TOMOGRAPHY; GLUCOSE-UTILIZATION; LIVER METASTASES; CARDIAC PET; QUANTIFICATION; WATER; (H2O)-O-15; BRAIN AB PET was used to measure tumor blood flow, which is potentially valuable for diagnosis and assessing the effects of therapy. To help visualize regional differences in blood flow and to improve the accuracy of region-of-interest placement, a parametric imaging approach was developed and compared with the standard region-of-interest method. Methods: Five patients with renal cell metastases in the thorax were studied using [O-15]water and dynamic PET. To assess the reproducibility of the blood flow measurements, multiple water studies were performed on each patient. Model fitting was done on a pixel-by-pixel basis using several different formulations of the standard single-compartment model. Results: The tumors studied spanned a wide range of blood flows, varying from 0.4 to 4.2 mL/min/g. These values were generally high compared with those of most other tissues, which meant that the tumors could be readily identified in parametric images of flow. The different model formulations produced images with different characteristics, and no model was entirely valid throughout the field of view. Although tumor blood flow measured from the parametric images was largely unbiased with respect to a standard regional method, large errors were observed with certain models in regions of low flow. The most robust model throughout the field of view had only 1 free parameter and, compared with a regional method, gave rise to a flow bias of 0.3% +/- 3.1% for tumor and 16% +/- 11% for low-flow soft tissue (muscle plus fat). With this model, tumor blood flow was measured with an SD of 7.6% +/- 4.0%. Conclusion: Parametric imaging provides a convenient way of visualizing regional changes in blood flow, which may be valuable in studies of tumor blood flow. C1 NIH, Ctr Clin, Bethesda, MD 20892 USA. NCI, NIH, Bethesda, MD 20892 USA. RP Bacharach, SL (reprint author), NIH, Ctr Clin, Rm 1C401,Bldg 10, Bethesda, MD 20892 USA. RI Carrasquillo, Jorge/E-7120-2010; Carson, Richard/H-3250-2011; OI Carson, Richard/0000-0002-9338-7966; Carrasquillo, Jorge/0000-0002-8513-5734 NR 34 TC 36 Z9 37 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD NOV PY 2000 VL 41 IS 11 BP 1784 EP 1792 PG 9 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 371XM UT WOS:000165204600006 PM 11079484 ER PT J AU Hoffman, JM Welsh-Bohmer, KA Hanson, M Crain, B Hulette, C Earl, N Coleman, RE AF Hoffman, JM Welsh-Bohmer, KA Hanson, M Crain, B Hulette, C Earl, N Coleman, RE TI FDG PET imaging in patients with pathologically verified dementia SO JOURNAL OF NUCLEAR MEDICINE LA English DT Article DE dementia; FDG PET; Alzheimer's disease ID POSITRON EMISSION TOMOGRAPHY; PROBABLE ALZHEIMERS-DISEASE; PROGRESSIVE SUPRANUCLEAR PALSY; DIMINISHED GLUCOSE-TRANSPORT; ADRDA WORK GROUP; DIFFERENTIAL-DIAGNOSIS; CLINICAL-DIAGNOSIS; CONSORTIUM; ESTABLISH; CRITERIA AB The purpose of this study was to confirm with pathologic verification 2 beliefs related to Alzheimer's disease (AD): (a) the long-standing impression that bilateral temporo-parietal hypometabolism, as noted on FDG PET imaging, is the metabolic abnormality associated with Alzheimer's disease (AD) and (b) that the sensitivity, specificity, and diagnostic accuracy of the metabolic pattern of bilateral temporo-parietal hypometabolism allows differentiation between other degenerative causes of dementia. Methods: Twenty two individuals (8 women, 14 men) with difficult-to-characterize memory loss or dementia (using standard clinical criteria), and who eventually received pathologic confirmation of diagnosis, were evaluated. FDG PET brain scans were obtained and visually graded by an experienced nuclear medicine physician as to the presence of classic bilateral temporo-parietal hypometabolism as seen in Alzheimer's type dementia. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of the metabolic pattern of bilateral temporo-parietal hypometabolism were determined using pathologic diagnosis as the gold standard. Results: The clinical diagnosis of possible or probable AD was determined as the primary cause of dementia in 12 patients. The sensitivity and specificity of the clinical diagnosis for probable AD were 63% and 100%, respectively. The sensitivity and specificity of the clinical diagnosis for possible and probable AD were 75% and 100%, respectively. The sensitivity, specificity, and diagnostic accuracy of bilateral temporo-parietal hypometabolism being associated with AD were 93%, 63%, and 82%, respectively. Conclusion: This study confirms that bilateral temporo-parietal hypometabolism is indeed the classic metabolic abnormality associated with AD. Furthermore, in individuals with dementia whose FDG PET scans indicated a metabolic pattern other than bilateral temporo-parietal hypometabolism, a cause of dementia other than AD should be suspected. These observations may be of clinical importance in differentiating dementia syndromes. The sensitivity, specificity, and diagnostic accuracy of FDG PET are acceptable as tests to be used in the evaluation of dementia and particularly to confirm the clinical suspicion of AD. C1 NCI, DCTT, DIP, EPN800, Bethesda, MD 20892 USA. Duke Univ, Joseph & Kathleen Bryan Alzheimers Dis Res Ctr, Durham, NC USA. Duke Univ, Med Ctr, Dept Psychiat & Behav Sci, Durham, NC USA. Duke Univ, Med Ctr, Dept Radiol, Durham, NC USA. Johns Hopkins Univ, Dept Pathol, Baltimore, MD USA. Duke Univ, Med Ctr, Dept Pathol, Durham, NC 27710 USA. RP Hoffman, JM (reprint author), NCI, DCTT, DIP, EPN800, 6130 Execut Blvd,MSC 7440, Bethesda, MD 20892 USA. FU NIA NIH HHS [P50 AG 05128-0751, 2P50 AG05128-07] NR 60 TC 185 Z9 192 U1 0 U2 7 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 NOV PY 2000 VL 41 IS 11 BP 1920 EP 1928 PG 9 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 371XM UT WOS:000165204600027 PM 11079505 ER PT J AU Michalowicz, BS Diehl, SR Gunsolley, JC Sparks, BS Brooks, CN Koertge, TE Califano, JV Burmeister, JA Schenkein, HA AF Michalowicz, BS Diehl, SR Gunsolley, JC Sparks, BS Brooks, CN Koertge, TE Califano, JV Burmeister, JA Schenkein, HA TI Evidence of a substantial genetic basis for risk of adult periodontitis SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE disease susceptibility; diseases in twins; twins; periodontitis/etiology; periodontitis/genetics ID EARLY-ONSET PERIODONTITIS; DWELLING OLDER ADULTS; TWIN DATA; ATTACHMENT LOSS; DISEASE; VARIANCE; GENOTYPE; ZYGOSITY AB Background: A few previous studies have suggested that risk for adult periodontitis (AP) has a genetic (heritable) component. We estimated genetic and environmental variances and heritability for gingivitis and adult periodontitis using data from twins reared together. Methods: One hundred seventeen (117) pairs of adult twins (64 monozygotic [MZ] and 53 dizygotic [DZ] pairs) were recruited. Probing depth (PD), attachment loss (AL), plaque, and gingivitis (GI) were assessed on all teeth by two examiners. Measurements were averaged over all sites, teeth, and examiners. Extent of disease in subjects was defined at four thresholds: the percentage of teeth with AL greater than or equal to2, AL greater than or equal to3, PD greater than or equal to4, or PD greater than or equal to5 mm. Genetic and environmental variances and heritability were estimated using path models with maximum likelihood estimation techniques. Results: MZ twins were more similar than DZ twins for all clinical measures. Statistically significant genetic variance was found for both the severity and extent of disease. AP was estimated to have approximately 50% heritability, which was unaltered following adjustments for behavioral variables including smoking. In contrast, while MZ twins were also more similar than DZ twins for gingivitis scores, there was no evidence of heritability for gingivitis after behavioral covariates such as utilization of dental care and smoking were incorporated into the analyses. Conclusions: These results confirm previous studies and indicate that approximately half of the variance in disease in the population is attributed to genetic variance. The basis for the heritability of periodontitis appears to be biological and not behavioral in nature. C1 Univ Minnesota, Sch Dent, Dept Prevent Sci, Minneapolis, MN 55455 USA. Natl Inst Dent & Craniofacial Res, Craniofacial Epidemiol & Genet Branch, Bethesda, MD USA. Virginia Commonwealth Univ, Sch Dent, Clin Res Ctr Periodontal Dis, Richmond, VA USA. Univ Maryland, Sch Dent, Dept Periodont, Baltimore, MD 21201 USA. RP Michalowicz, BS (reprint author), 7-368 Moos Tower,515 Delaware St SE, Minneapolis, MN 55455 USA. FU NIDCR NIH HHS [DE00622, P30DE09737, P50DE10703] NR 37 TC 260 Z9 271 U1 3 U2 14 PU AMER ACAD PERIODONTOLOGY PI CHICAGO PA 737 NORTH MICHIGAN AVENUE, SUITE 800, CHICAGO, IL 60611-2690 USA SN 0022-3492 J9 J PERIODONTOL JI J. Periodont. PD NOV PY 2000 VL 71 IS 11 BP 1699 EP 1707 DI 10.1902/jop.2000.71.11.1699 PG 9 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 411GQ UT WOS:000167490000005 PM 11128917 ER PT J AU Hasinoff, BB Abram, ME Chee, GL Huebner, E Byard, EH Barnabe, N Ferrans, VJ Yu, ZX Yalowich, JC AF Hasinoff, BB Abram, ME Chee, GL Huebner, E Byard, EH Barnabe, N Ferrans, VJ Yu, ZX Yalowich, JC TI The catalytic DNA topoisomerase II inhibitor dexrazoxane (ICRF-187) induces endopolyploidy in Chinese hamster ovary cells SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID MEDIATED GROWTH-INHIBITION; CYCLE PROGRESSION; MAMMALIAN-CELLS; INDUCTION; PROTEIN; BISDIOXOPIPERAZINES; POLYPLOIDIZATION; LINE; DIFFERENTIATION; MICROTUBULES AB The bisdioxopiperazines, including dexrazoxane (ICRF-187), are catalytic or noncleavable complex-forming inhibitors of DNA topoisomerase II that do not produce DNA strand breaks. In this study we show that dexrazoxane inhibits the division of Chinese hamster ovary (CHO) cells resulting in marked increases in cell size (up to 80 mum in diameter), volume (up to 150-fold greater), and ploidy (as high as 32N). This last result indicates that the dexrazoxane-induced DNA reduplication was restricted to once per cell cycle. Kinetic analysis of the flow cytometry data indicated that the conversion between successively higher ploidy levels was progressively slowed at longer times of exposure to dexrazoxane. Both the protein and DNA content of dexrazoxane-treated CHO cells increased linearly over time in the same proportion. Light and electron microscopic studies of dexrazoxane-treated cells showed ring-like multilobulated nuclei. Immunohistochemical staining of dexrazoxane-treated cells showed that F-actin and acetylated alpha -tubulin were present in large, highly organized networks. Immunohistochemical staining of the dexrazoxane-treated CHO cells also showed that the topoisomerase II a colocalized with the DNA of the multilobulated nuclei. Staining of gamma -tubulin revealed that the dexrazoxane-treated cells contained multiple centrosomes, indicating that dexrazoxane prevents cytokinesis but not centrosome reduplication. It is concluded that dexrazoxane inhibits CHO cytokinesis in cells by virtue of its ability to inhibit topoisomerase II. C1 Univ Manitoba, Fac Pharm, Winnipeg, MB R3T 2N2, Canada. Univ Manitoba, Dept Zool, Winnipeg, MB R3T 2N2, Canada. Univ Winnipeg, Dept Biol, Winnipeg, MB R3B 2E9, Canada. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. Univ Pittsburgh, Sch Med, Dept Pharmacol, Pittsburgh, PA 15261 USA. Pittsburgh Canc Inst, Pittsburgh, PA 15213 USA. RP Hasinoff, BB (reprint author), Univ Manitoba, Fac Pharm, Winnipeg, MB R3T 2N2, Canada. FU NCI NIH HHS [CA77468, CA74972] NR 42 TC 17 Z9 17 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 NOV PY 2000 VL 295 IS 2 BP 474 EP 483 PG 10 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 366TZ UT WOS:000090021800006 PM 11046078 ER PT J AU Hoare, SRJ Usdin, TB AF Hoare, SRJ Usdin, TB TI Tuberoinfundibular peptide (7-39) [TIP(7-39)], a novel, selective, high-affinity antagonist for the parathyroid hormone-1 receptor with no detectable agonist activity SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID TERMINAL REGION; PTH2 RECEPTOR; BINDING; INVIVO; HYPERCALCEMIA; PROTEIN; RAT; CELLS; IDENTIFICATION; EXPRESSION AB The parathyroid hormone (PTH)-1 receptor mediates the pathophysiological effects of PTH in hyperparathyroidism and PTH-related protein (PTHrP) in humoral hypercalcemia of malignancy. A PTH1 receptor antagonist may be of therapeutic utility in these disorders. We recently identified a novel antagonist, tuberoinfundibular peptide (7-39) [TIP( 7-39)], derived from the likely endogenous ligand for the PTH2 receptor TIP39. In this study its in vitro profile is evaluated and compared with that of [D-Trp(12),Tyr(34)]bPTH(7-34) and PTHrP(7-34), representing the two previously known structural classes of PTH1 receptor antagonists. TIP(7-39) binds with higher affinity (6.2 nM) to the PTH1 receptor than [D-Trp(12),Tyr(34)]bPTH(7-34) (45 nM) and PTHrP(7-34) (65 nM) and displays a 5.5-fold greater PTH1/PTH2 receptor selectivity. TIP(7-39) does not stimulate cAMP accumulation via the PTH1 receptor [in a sensitive assay that detects the activity of the weak partial agonist [Nle(8,18),Tyr(34)]bPTH(3-34)] and does not increase intracellular calcium. Schild analysis for TIP(7-39) was consistent with purely competitive antagonism of PTH(1-34)'s stimulation of cAMP accumulation (slope = 0.99 +/- 0.24). The pK(B) for TIP(7-39) (7.1 +/- 0.3) was higher than that for [D-Trp(12),Tyr(34)]bPTH(7-34) (6.5 +/- 0.0) and PTHrP(7-34) (6.0 +/- 0.1). Binding of I-125-TIP(7-39) to the PTH1 receptor could be measured (K-D = 1.3 +/- 0.1 nM, B-max = 1.3 +/- 0.1 pmol/ mg), whereas binding of I-125[Nle(8,18),D-Trp(12),Tyr(34)]bPTH(7-34) could not be detected. Kinetic analysis indicated that I-125-TIP(7-39) dissociates much more slowly (t(1/2) = 14 min) than [D-Trp(12),Tyr(34)]bPTH(7-34) (13 s) and PTHrP(7-34) (9 s). The novel antagonist TIP(7-39) therefore displays a more favorable in vitro pharmacological profile than antagonists derived from PTH and PTHrP and may be useful for demonstrating the utility of PTH1 receptor antagonism in the treatment of hypercalcemia. C1 NIMH, Cell Biol Unit, Genet Lab, NIH, Bethesda, MD 20892 USA. RP Usdin, TB (reprint author), NIMH, Cell Biol Unit, Genet Lab, NIH, Rm 3D06,Bldg 36,36 Convent Dr,MSC4094, Bethesda, MD 20892 USA. NR 40 TC 22 Z9 23 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 NOV PY 2000 VL 295 IS 2 BP 761 EP 770 PG 10 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 366TZ UT WOS:000090021800044 PM 11046116 ER PT J AU Diri, E Lipsky, PE Berggren, RE AF Diri, E Lipsky, PE Berggren, RE TI Emergence of systemic lupus erythematosus after initiation of highly active antiretroviral therapy for human immunodeficiency virus infection SO JOURNAL OF RHEUMATOLOGY LA English DT Article DE autoimmunity; lupus; human immunodeficiency virus; immune reconstitution disease; antiretroviral therapy ID CONTROLLED TRIAL; HIV; DISEASE; PATIENT; COEXISTENCE; RITONAVIR AB Systemic lupus erythematosus (SLE) may be influenced by human immunodeficiency virus type-1 (HIV) infection. It has been suggested that the immunosuppression resulting from HIV infection can prevent the emergence of SLE. There appear to be fewer cases of SLE in the HIV infected population than would be predicted, based on the overall incidence of SLE. We describe a person with HIV infection who developed SLE after the initiation of highly active antiretroviral therapy. C1 Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Div Rheumat Dis, Dallas, TX 75390 USA. NIAMSD, NIH, Bethesda, MD 20892 USA. RP Berggren, RE (reprint author), Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis, 5323 Harry Hines Blvd, Dallas, TX 75390 USA. FU NIAID NIH HHS [1R21AI42642] NR 21 TC 48 Z9 49 U1 0 U2 0 PU J RHEUMATOL PUBL CO PI TORONTO PA 920 YONGE ST, SUITE 115, TORONTO, ONTARIO M4W 3C7, CANADA SN 0315-162X J9 J RHEUMATOL JI J. Rheumatol. PD NOV PY 2000 VL 27 IS 11 BP 2711 EP 2714 PG 4 WC Rheumatology SC Rheumatology GA 371XK UT WOS:000165204400033 PM 11093459 ER PT J AU Hickel, KW AF Hickel, KW TI "Justice and the highest kind of equality require discrimination": Citizenship, dependency and conscription in the South, 1917-1919 SO JOURNAL OF SOUTHERN HISTORY LA English DT Article C1 Natl Lib Med, Bethesda, MD 20209 USA. RP Hickel, KW (reprint author), Natl Lib Med, Bethesda, MD 20209 USA. NR 25 TC 7 Z9 7 U1 0 U2 0 PU SOUTHERN HISTORICAL ASSOC PI ATHENS PA UNIV GEORGIA, HISTORY DEPT, ATHENS, GA 30602 USA SN 0022-4642 J9 J SOUTHERN HIST JI J. South. Hist. PD NOV PY 2000 VL 66 IS 4 BP 749 EP 780 DI 10.2307/2588010 PG 32 WC History SC History GA 373PA UT WOS:000165296800002 ER PT J AU Borkowf, CB AF Borkowf, CB TI On multidimensional contingency tables with categories defined by the empirical quantiles of the marginal data SO JOURNAL OF STATISTICAL PLANNING AND INFERENCE LA English DT Article DE empirical multivariate quantile-partitioned (EMQP) distribution; intraclass correlation; nonparametric statistics; nutritional epidemiology; quantiles; ranks ID DIETARY ASSESSMENT INSTRUMENTS; FOOD FREQUENCY QUESTIONNAIRE; REPRODUCIBILITY; VALIDITY; DISTRIBUTIONS AB Epidemiologists sometimes collect multivariate continuous data on a number of subjects and then wish to analyze the relationships and the agreement between these measurements. They may choose to compute the empirical quantiles (e.g., medians or quintiles) of each of these measurements and use these quantities to define the marginal categories of a multidimensional contingency table. Such contingency tables have the empirical multivariate quantile-partitioned (EMQP) distribution. an extension of the bivariate case studied by Borkowf et al. (1997, Biometrics 53, 1054-1069). In this paper, we present the general asymptotic theory for the EMQP distribution and nonparametric large sample procedures to estimate the variances of such tables from observed multivariate data. We define several statistics of interest to epidemiologists that can be calculated from EMQP tables, including the intraclass correlation, and present numerical results for these statistics where the original data come from certain underlying trivariate distributions. We also discuss the relationship of the EMQP distribution to other distributions commonly used in categorical data analysis. In addition, we apply EMQP methods to analyze an example from nutritional epidemiology. Finally, we discuss how EMQP theory and methods can unify the distribution theory and estimation procedures for diverse nonparametric statistics calculated from multivariate ranks or quantile-categories. (C) 2000 Elsevier Science B.V. All rights reserved. MSC: 62G20; 62G30; 92C60. C1 NCI, Div Clin Sci, Canc Prevent Studies Branch, Bethesda, MD 20892 USA. NR 22 TC 3 Z9 3 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-3758 J9 J STAT PLAN INFER JI J. Stat. Plan. Infer. PD NOV 1 PY 2000 VL 91 IS 1 BP 33 EP 51 DI 10.1016/S0378-3758(00)00127-0 PG 19 WC Statistics & Probability SC Mathematics GA 363GZ UT WOS:000089828400003 ER PT J AU Adachi, M Sugawara, M Iwasa, KH AF Adachi, M Sugawara, M Iwasa, KH TI Effect of turgor pressure on outer hair cell motility SO JOURNAL OF THE ACOUSTICAL SOCIETY OF AMERICA LA English DT Article ID COCHLEAR AMPLIFIER; FORCE GENERATION; MEMBRANE; TRANSDUCTION; CAPACITANCE; MOTOR AB The motor in the outer hair cell converts electrical energy directly into mechanical energy. There are two possible mechanisms for such a motor: one depends on changes in the membrane area ("area motor") and the other on changes in the stiffness ("stiffness motor"). These two mechanisms, which are not mutually exclusive, give different predictions on turgor pressure dependence of the amplitude. It was found that an increased pressure shifts the voltage dependence but does not change the amplitude of length changes. This observation is incompatible with a pure stiffness motor model. It was also confirmed that length changes are closely related to charge movements by monitoring the membrane capacitance. It can be concluded that hair cell motility is primarily based on area changes, and not changes in the elastic moduli. [S0001-4966(00)02811-3]. C1 Natl Inst Deafness & Other Commun Disorders, Lab Cellular Biol, Biophys Sect, NIH, Bethesda, MD 20892 USA. RP Iwasa, KH (reprint author), Natl Inst Deafness & Other Commun Disorders, Lab Cellular Biol, Biophys Sect, NIH, Bethesda, MD 20892 USA. OI Iwasa, Kuni/0000-0002-9397-7704 NR 30 TC 13 Z9 13 U1 0 U2 0 PU AMER INST PHYSICS PI MELVILLE PA 2 HUNTINGTON QUADRANGLE, STE 1NO1, MELVILLE, NY 11747-4501 USA SN 0001-4966 J9 J ACOUST SOC AM JI J. Acoust. Soc. Am. PD NOV PY 2000 VL 108 IS 5 BP 2299 EP 2306 DI 10.1121/1.1314396 PN 1 PG 8 WC Acoustics; Audiology & Speech-Language Pathology SC Acoustics; Audiology & Speech-Language Pathology GA 372GR UT WOS:000165226400035 PM 11108370 ER PT J AU Prasad, A Husain, S Schenke, W Mincemoyer, R Epstein, N Quyyumi, AA AF Prasad, A Husain, S Schenke, W Mincemoyer, R Epstein, N Quyyumi, AA TI Contribution of bradykinin receptor dysfunction to abnormal coronary vasomotion in humans SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID ENDOTHELIUM-DEPENDENT RELAXATIONS; ANGIOTENSIN-CONVERTING ENZYME; SENSITIVE G-PROTEIN; ARTERIES IN-VIVO; NITRIC-OXIDE; INDUCED VASODILATION; RISK-FACTORS; INHIBITION; ACETYLCHOLINE; HYPERCHOLESTEROLEMIA AB OBJECTIVES The aim of our study was to investigate coronary vascular kinin receptor function in patients with atherosclerosis or its risk factors. BACKGROUND Although acetylcholine (ACH) is used as a probe for testing vascular function in vivo, endogenous bradykinin (BK) regulates resting and flow-mediated epicardial tone. METHODS In 53 patients with mild atherosclerosis or its risk factors and 9 control subjects, endothelium-dependent vasomotion was tested with intracoronary ACH (30 mug/min) and BK (62.5 ng/min and 4 mug/min), and endothelium independent function with sodium nitroprusside. Metabolic vasodilation was assessed during cardiac pacing (n = 19). Correlation with serum angiotensin-converting enzyme (ACE) levels and the ACE insertion/deletion genotype was performed. RESULTS There was progressive impairment in AGH-mediated microvascular dilation with increasing numbers of risk factors (p = 0.025, analysis of variance). By contrast, BK- and sodium nitroprusside-mediated microvascular dilation was similar in all groups. Similarly, there was no correlation between epicardial coronary responses to ACH and BK; segments that constricted or dilated with ACH had similar dilator responses with BK. Bradykinin, but not ACH-mediated vasomotion, was depressed in epicardial segments that constricted with pacing. Finally, epicardial BK responses were depressed in patients with high ACE levels and in those with the ACE DD genotype. CONCLUSIONS Endothelial dysfunction in atherosclerosis appears to be receptor-specific, involving the muscarinic receptor with relative sparing of the kinin receptor pathways. Abnormal reactivity of epicardial coronary arteries during physiologic stress is better represented by BK and not by ACH responses. Bradykinin activity and, hence, physiologic coronary vasomotion appears to be influenced by serum ACE levels and the ACE insertion/deletion genotype. (C) 2000 by the American College of Cardiology. C1 NHLBI, NIH, Cardiol Branch, Bethesda, MD 20892 USA. RP Quyyumi, AA (reprint author), NHLBI, NIH, Cardiol Branch, Bldg 10,Rm 7B15,10 Ctr Dr,MSC 1650, Bethesda, MD 20892 USA. NR 30 TC 19 Z9 19 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD NOV 1 PY 2000 VL 36 IS 5 BP 1467 EP 1473 DI 10.1016/S0735-1097(00)00892-5 PG 7 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 371GY UT WOS:000165171500003 PM 11079644 ER PT J AU Cardillo, C Kilcoyne, CM Cannon, RO Panza, JA AF Cardillo, C Kilcoyne, CM Cannon, RO Panza, JA TI Increased activity of endogenous endothelin in patients with hypercholesterolemia SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID CONVERTING ENZYME-INHIBITION; NITRIC-OXIDE; RECEPTOR ANTAGONISM; GENE-EXPRESSION; BLOOD-PRESSURE; VASCULAR TONE; ET(A); IMMUNOREACTIVITY; VESSELS; POTENT AB OBJECTIVE We sought to assess the activity of endogenous endothelin-l (ET-1) in hypercholesterolemic patients using antagonists of ET-1 receptors. BACKGROUND Endothelial dysfunction in hypercholesterolemic patients may contribute to their risk of premature atherosclerosis. Endothelin, a peptide released by endothelial cells, may be involved in this process by activating smooth muscle cell. mitogenesis and leukocyte adhesion. METHODS Forearm blood flow (FBF) responses (strain-gauge plethysmography) to intra-arterial infusion of a selective blocker of ETA receptors (BQ-123) and, on a separate occasion, to ET-1 were measured in 12 hypercholesterolemic patients and 12 normal control subjects. In addition, on a different day, six hypercholesterolemic patients received co-infusion of BQ-123 and BQ-788 (a selective blocker of ETB receptors). RESULTS In normal subjects, BQ-123 did not significantly modify FBF from baseline (p = 0.78); however, in hypercholesterolemic patients, BQ-123 administration resulted in a significant vasodilator response (p ( 0.001). Administration of exogenous ET-1 resulted in similar vasoconstrictor responses in patients (37%) and control subjects (35%) (p = 0.83). In hypercholesterolemic patients, the vasodilator response to selective ETA blockade was reversed by nonselective blockade of ET-1 receptors obtained by co-infusion of BQ-123 and BQ-788. CONCLUSIONS The vascular activity of endogenous ET-1 is enhanced in hypercholesterolemic patients, whereas their sensitivity to exogenous ET-1 is unchanged. These findings suggest increased production of ET-1, which may participate in the pathophysiology of vascular disease characteristic of hypercholesterolemia. (C) 2000 by the American College of Cardiology. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Panza, JA (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,10 Ctr Dr,MSC 1650,Room 7B-15, Bethesda, MD 20892 USA. NR 32 TC 51 Z9 54 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD NOV 1 PY 2000 VL 36 IS 5 BP 1483 EP 1488 DI 10.1016/S0735-1097(00)00910-4 PG 6 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 371GY UT WOS:000165171500005 PM 11079646 ER PT J AU Prasad, A Narayanan, S Waclawiw, MA Epstein, N Quyyumi, AA AF Prasad, A Narayanan, S Waclawiw, MA Epstein, N Quyyumi, AA TI The insertion/deletion polymorphism of the angiotensin-converting enzyme gene determines coronary vascular tone and nitric oxide activity SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID ISCHEMIC-HEART-DISEASE; PLASMA ACE LEVELS; DELETION POLYMORPHISM; MYOCARDIAL-INFARCTION; LINKAGE ANALYSIS; RISK-FACTORS; GENOTYPE; ARTERIES; RAT; VASOCONSTRICTION AB OBJECTIVES We investigated whether the insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene modulates vasomotor tone and endothelial function. BACKGROUND The deletion allele of the ACE I/D polymorphism has been associated with increased incidence of cardiovascular pathology. The risk is synergistically increased in patients who also possess the C allele at position 1,166 of the angiotensin type I (AT1) receptor gene. METHODS In 177 patients with coronary atherosclerosis or its risk factors, we investigated endothelial function with intracoronary acetylcholine (ACH), endothelium-independent smooth muscle function with sodium nitroprusside (SNP) and basal nitric oxide activity with L-N-G monomethyl arginine. RESULTS Compared with ACE II genotype, patients with the ACE DD genotype had lower coronary microvascular and epicardial responses with SNP (coronary blood flow increase 196 +/- 26% vs. 121 +/- 11%, p = 0.003, and diameter increase 21.9 +/- 2% vs. 17 +/- 1%, p = 0.03, ACE II vs. DD, respectively). L-N-G monomethyl arginine induced greater constriction in patients with the ACE DD compared with ACE II genotype (coronary blood flow -10 +/- 4% vs. 11 +/- 5%, p = 0.003, ACE DD vs. II and diameter constriction -6.3 +/- 1.2% vs. -1.9 +/- 1.2%, p = 0.01, respectively, in patients with atherosclerosis). No difference in ACH mediated vasomotion was detected between the three ACE genotypes. The AT1 receptor polymorphism did not influence responses to tither SNP or ACH. CONCLUSIONS Patients possessing the D allele of the ACE gene have increased vascular smooth muscle tone. The enhanced tone appears tu be counterbalanced by an increase in basal nitric oxide activity in patients with atherosclerosis. (C) 2000 by the American College of Cardiology. C1 NHLBI, NIH, Cardiol Branch, Bethesda, MD 20892 USA. RP Quyyumi, AA (reprint author), NHLBI, NIH, Cardiol Branch, Bldg 10,Rm 7B15,10 Ctr Dr,MSC 1650, Bethesda, MD 20892 USA. NR 38 TC 29 Z9 30 U1 0 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD NOV 1 PY 2000 VL 36 IS 5 BP 1579 EP 1586 DI 10.1016/S0735-1097(00)00902-5 PG 8 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 371GY UT WOS:000165171500020 PM 11079661 ER PT J AU Libutti, SK Bartlett, DL Fraker, DL Alexander, HR AF Libutti, SK Bartlett, DL Fraker, DL Alexander, HR TI Technique and results of hyperthermic isolated hepatic perfusion with tumor necrosis factor and melphalan for the treatment of unresectable hepatic malignancies SO JOURNAL OF THE AMERICAN COLLEGE OF SURGEONS LA English DT Article ID ISOLATED LIVER PERFUSION; ISOLATED LIMB PERFUSION; COLORECTAL-CANCER; FACTOR-ALPHA; METASTASES; MELANOMA; CHEMOTHERAPY; EXPERIENCE; CARCINOMA; TNF AB Background: For a variety of histologies, the liver represents the only or the dominant site of metastatic disease. Regional treatment of the liver has the theoretic advantage of maximizing drug delivery while minimizing systemic toxicity. This article describes the technique of isolated hepatic perfusion using tumor necrosis factor and melphalan under conditions of moderate hyperthermia for the treatment of unresectable liver tumors. Study Design: Fifty patients with biopsy-proved unresectable primary or metastatic cancer to the liver were treated. Isolated hepatic perfusion was performed for 60 minutes under conditions of moderate hyperthermia during a laparotomy with inflow through the gastroduodenal artery and outflow through an isolated segment of inferior vena cava. During isolated hepatic perfusion portal and infrahepatic blood flow were shunted externally by a centrifugal pump to the axillary vein. Complete vascular isolation was confirmed intraoperatively using a continuous I-131-labeled serum albumin leak monitoring system. Operative and perfusion parameters were recorded. Results: By using a standard operative technique to achieve complete vascular isolation of the liver during perfusion, there was no leak of perfusate detected in 48 of 50 patients as determined by the continuous leak monitoring system and absence of detectable systemic tumor necrosis factor levels. Operating time, transfusion requirements, and blood loss were within the range expected for a major operative procedure. Stable hemodynamic and perfusion parameters were achieved consistently and all patients successfully completed the 60-minute perfusion. Two patients (4%) died as a result of treatment and significant tumor regression was observed in 75%. Conclusions: Isolated hepatic perfusion is a technique that can be used to deliver high doses of chemotherapy or biologic therapy regionally and without systemic exposure. By using a standard operative technique, continuous intraoperative leak monitoring, and an external veno-veno bypass circuit, this procedure can be done safely and with acceptable morbidity and mortality. This article demonstrates that sustained and complete vascular isolation of the liver can be achieved and indicates further study is warranted to better define the role of isolated hepatic perfusion in the treatment of unresectable liver tumors. (J Am Coll Surg 2000;191: 519-530. (C) 2000 by the American College of Surgeons). C1 NCI, Surg Metab Sect, Surg Branch, NIH, Bethesda, MD 20892 USA. Univ Penn, Dept Surg, Philadelphia, PA 19104 USA. RP Alexander, HR (reprint author), NCI, Surg Metab Sect, Surg Branch, NIH, Bldg 10,Room 2B07, Bethesda, MD 20892 USA. NR 30 TC 44 Z9 46 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1072-7515 J9 J AM COLL SURGEONS JI J. Am. Coll. Surg. PD NOV PY 2000 VL 191 IS 5 BP 519 EP 530 DI 10.1016/S1072-7515(00)00733-X PG 12 WC Surgery SC Surgery GA 369KG UT WOS:000165064000006 PM 11085732 ER PT J AU Penninx, BWJH Guralnik, JM Bandeen-Roche, K Kasper, JD Simonsick, EM Ferrucci, L Fried, LP AF Penninx, BWJH Guralnik, JM Bandeen-Roche, K Kasper, JD Simonsick, EM Ferrucci, L Fried, LP TI The protective effect of emotional vitality on adverse health outcomes in disabled older women SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Article DE emotional vitality; psychological well-being; disability; mortality; longitudinal study ID DEPRESSIVE SYMPTOMS; MORTALITY; DISABILITY; AGE; POPULATION AB OBJECTIVE: Although the adverse physical health consequences of negative emotions have been studied extensively, much less is known about the potential impact of positive emotions. This study examines whether emotional vitality protects against progression of disability and mortality in disabled older women. DESIGN: A community-based study, The Women's Health and Aging Study. PARTICIPANTS: A total of 1002 moderately to severely disabled women aged 65 years and older living in the community. MEASUREMENTS: Emotional vitality was defined as having a high sense of personal mastery, being happy, and having low depressive symptomatology and anxiety. The onset of new disability was determined by semiannual assessments of disability in performing activities of daily living (ADLs), walking across a room, walking 1/4 mile, and lifting/carrying 10 pounds. Mortality status was determined by proxy interviews and linkage with death certificates. Survival analyses with time to onset of specific disabilities (among those not disabled at baseline) and time to mortality were performed and adjusted for age, baseline level of difficulty, physical performance, and chronic conditions. RESULTS: Three hundred fifty-one of the 1002 older disabled women studied were emotionally vital. Among women without the specific disability at baseline, emotional vitality was associated with a significantly decreased risk for incident disability performing ADLs (RR = 0.81, 95% CI = 0.66-0.99), for incident disability walking one-quarter mile (RR = 0.73, 95% CI = 0.59-0.92), and for incident disability lifting/carrying 10 pounds (RR = 0.77, 95% CI = 0.63-0.95). Emotional vitality was also associated with a lower risk of dying (RR = 0.56, 95% CI = 0.39-0.80). These results were not simply caused by the absence of depression since protective health effects remained when emotionally vital women were compared with 334 women who were not emotionally vital and not depressed. CONCLUSIONS: Emotional vitality in older disabled women reduces the risk for subsequent new disability and mortality. Our findings suggest that positive emotions can protect older persons against adverse health outcomes. C1 Wake Forest Univ, Sch Med, Sticht Ctr Aging, Winston Salem, NC 27157 USA. NIA, Epidemiol Demog & Biomety Program, Bethesda, MD 20892 USA. Johns Hopkins Hyg & Publ Hlth, Dept Biostat, Baltimore, MD USA. Ist Nazl Ric & Cura Per Gli Anziani, Geriatr Dept 1, Florence, Italy. Johns Hopkins Med Inst, Dept Med, Baltimore, MD 21205 USA. Johns Hopkins Med Inst, Dept Epidemiol, Baltimore, MD 21205 USA. RP Penninx, BWJH (reprint author), Wake Forest Univ, Sch Med, Sticht Ctr Aging, Med Ctr Blvd, Winston Salem, NC 27157 USA. FU NIA NIH HHS [AG-09834, N01-AG-1-2112] NR 27 TC 75 Z9 77 U1 1 U2 10 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD NOV PY 2000 VL 48 IS 11 BP 1359 EP 1366 PG 8 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373EY UT WOS:000165276500001 PM 11083309 ER PT J AU Reuben, DB Ferrucci, L Wallace, R Tracy, RP Corti, C Heimovitz, H Harris, TB AF Reuben, DB Ferrucci, L Wallace, R Tracy, RP Corti, C Heimovitz, H Harris, TB TI The prognostic value of serum albumin in healthy older persons with low and high serum interleukin-6 (IL-6) levels SO JOURNAL OF THE AMERICAN GERIATRICS SOCIETY LA English DT Article DE inflammation; cytokines; albumin; interleukin-6; malnutrition ID LOWER-EXTREMITY FUNCTION; MORTALITY; DISABILITY; ASSOCIATION; PREDICTOR; HYPOCHOLESTEROLEMIA; HYPOALBUMINEMIA AB OBJECTIVE: We sought to determine the prognostic value of serum albumin for 4-year mortality among high-functioning persons who did or did not have evidence of inflammation as indicated by high interleukin-6 (IL-6) levels. DESIGN: We used a case-cohort design of healthy, nondisabled older persons who had serum albumin and plasma IL-6 measured at baseline. Crude and multiply adjusted (for sociodemographics and chronic diseases) proportional hazards models were used to identify the effect of baseline levels of serum albumin level on 4-year mortality among those with higher and lower levels of IL-6. RESULTS: Among subjects without evidence of IL-6-mediated inflammation (IL-6 < 3.20 pg/mL), having a lower (4.4 g/dL) albumin level was associated with a multiply adjusted relative risk of 2.1 for 4-year mortality compared with those with higher albumin. In the presence of inflammation (IL-6 greater than or equal to 3.20 pg/mL), higher and lower serum albumin levels had similar risks (adjusted relative risks 4.0 and 3.8, respectively) compared with the referent group (higher albumin and low IL-6). CONCLUSIONS: High serum albumin has a protective effect in healthy older persons who do not have evidence of cytokine-mediated inflammation. This protective effect is not conferred in presence of inflammation. The mechanisms by which inflammation eliminates the protective effect of high albumin remain to be determined. C1 Univ Calif Los Angeles, Sch Med, Multicampus Program Geriatr Med & Gerontol, Los Angeles, CA 90095 USA. Italian Natl Res Council Aging, Florence, Italy. Univ Iowa, Coll Med, Iowa City, IA USA. Univ Vermont, Burlington, VT USA. Univ Padua, Padua, Italy. Sytel, Bethesda, MD USA. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. RP Reuben, DB (reprint author), Univ Calif Los Angeles, Sch Med, Multicampus Program Geriatr Med & Gerontol, 10945 Le Conte Ave,Suite 2339, Los Angeles, CA 90095 USA. FU NIA NIH HHS [AG10415-01] NR 15 TC 36 Z9 38 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0002-8614 J9 J AM GERIATR SOC JI J. Am. Geriatr. Soc. PD NOV PY 2000 VL 48 IS 11 BP 1404 EP 1407 PG 4 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373EY UT WOS:000165276500007 PM 11083315 ER PT J AU Marzilli, LA Golden, TR Cotter, RJ Woods, AS AF Marzilli, LA Golden, TR Cotter, RJ Woods, AS TI Peptide sequence information derived by Pronase Digestion and ammonium sulfate in-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry SO JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY LA English DT Article ID PARTIAL ACID-HYDROLYSIS; DESORPTION IONIZATION; CARBOXYPEPTIDASE-Y; PROTEINS; IDENTIFICATION; FRAGMENTATION; EXOPEPTIDASES; DEGRADATION; REFLECTRON; MALDI AB We present the use of Pronase digestion and in-source decay in the presence of ammonium sulfate as complementary techniques to confirm the amino acid sequence of a peptide. Pronase, a commercial preparation from Steptomyces griseus, is a combination of proteolytic enzymes. It produces carboxypeptidase and aminopeptidase ladders using a single Pronase digestion and represents an inexpensive, nonspecific, and fast supplement to traditional sequencing enzymes. However, N-terminal peptidase activity appears dependent on the terminal amino acid residue. We also introduce the use of saturated ammonium sulfate as an "on-slide" sample additive to promote in-source fragmentation of peptides. Use of saturated ammonium sulfate resulted in a simple way to increase peptide backbone fragmentation and essentially produced either a c(n) or y(n) ion series. Together these techniques provide useful supplements to existing methods for peptide sequence information, (C) 2000 American Society for Mass Spectrometry. C1 Johns Hopkins Univ, Sch Med, Dept Pharmacol & Mol Sci, Baltimore, MD 21218 USA. RP NIDA, Intramural Program, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. EM awoods@intra.nida.nih.gov FU NIGMS NIH HHS [GM 54882] NR 38 TC 15 Z9 16 U1 1 U2 3 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 1044-0305 EI 1879-1123 J9 J AM SOC MASS SPECTR JI J. Am. Soc. Mass Spectrom. PD NOV PY 2000 VL 11 IS 11 BP 1000 EP 1008 DI 10.1016/S1044-0305(00)00170-7 PG 9 WC Biochemical Research Methods; Chemistry, Analytical; Chemistry, Physical; Spectroscopy SC Biochemistry & Molecular Biology; Chemistry; Spectroscopy GA 367WR UT WOS:000090084900009 PM 11073263 ER PT J AU Lemley, KV Blouch, K Abdullah, I Boothroyd, DB Bennett, PH Myers, BD Nelson, RG AF Lemley, KV Blouch, K Abdullah, I Boothroyd, DB Bennett, PH Myers, BD Nelson, RG TI Glomerular permselectivity at the onset of nephropathy in type 2 diabetes mellitus SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article ID PIMA-INDIANS; RENAL-DISEASE; OVERT NEPHROPATHY; ALBUMIN EXCRETION; SIZE-SELECTIVITY; URINARY ALBUMIN; PROTEINURIA; PROGRESSION; CHARGE; MICROALBUMINURIA AB The development of microalbuminuria in individuals with type 2 diabetes mellitus is associated with a 10-fold increase in the risk of progression to overt nephropathy and eventual end-stage renal failure, The present study reports long-term (up to 8 yr) follow-up of 43 Pima Indians with type 2 diabetes detected on screening to have microalbuminuria. The natural history of albuminuria in these individuals included progression to overt proteinuria (urinary albumin excretion greater than or equal to 300 mg/d) in half of the participants by 7 yr of follow-up. The size selectivity of the glomerular barrier was also investigated using dextran sieving and pore theory. Whereas a comparison group of macroalbuminuric Pima Indians had an excess of large pores that, served as a macromolecular "shunt," individuals with microalbuminuria had a shunt size no different from long-term diabetic, normoalbuminuric control subjects. An abrupt transition from little or no relationship to a highly significant and positive relationship between increasing albuminuria and shunt size occurred at an albumin-to-creatinine ratio of approximately 3000 mg/g. Shunt size in macroalbuminuric individuals correlated with the extent of foot process broadening. Podocyte foot processes in microalbuminuric participants were not different from those in control subjects. In conclusion, although microalbuminuria in type 2 diabetic Pima Indians often heralds progressive glomerular injury, it is not the result of defects in the size permselectivity of the glomerular barrier but rather of changes in either glomerular charge selectivity or tubular handling of filtered proteins or of a combination of these two factors. C1 Stanford Univ, Med Ctr, Div Pediat Nephrol, Sch Med, Stanford, CA 94305 USA. Stanford Univ, Sch Med, Div Nephrol, Stanford, CA 94305 USA. Stanford Univ, Sch Med, Dept Stat, Stanford, CA 94305 USA. NIDDKD, Phoenix Epidemiol & Clin Res Branch, Phoenix, AZ 85016 USA. RP Lemley, KV (reprint author), Stanford Univ, Med Ctr, Div Pediat Nephrol, Sch Med, Room G306, Stanford, CA 94305 USA. RI Nelson, Robert/B-1470-2012 FU NCRR NIH HHS [M01-RR00070]; NIDDK NIH HHS [1R01-DK54600] NR 29 TC 59 Z9 63 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 NOV PY 2000 VL 11 IS 11 BP 2095 EP 2105 PG 11 WC Urology & Nephrology SC Urology & Nephrology GA 368VQ UT WOS:000090138500017 PM 11053486 ER PT J AU McKee, JA Kumar, S Ecelbarger, CA Fernandez-Llama, P Terris, J Knepper, MA AF McKee, JA Kumar, S Ecelbarger, CA Fernandez-Llama, P Terris, J Knepper, MA TI Detection of Na+ transporter proteins in urine SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article ID DIABETES-INSIPIDUS; AQUAPORIN-2; EXCRETION; VASOPRESSIN; CHANNEL; RAT; EXPRESSION AB Previous studies have established that the vasopressin-regulated water channel of the collecting duct, aquaporin-2, is excreted in the urine, providing a means for assessment of regulation and dysregulation of aquaporin-2 in humans. This article addresses the hypothesis that membrane transporters from upstream nephron segments are normally detectable in urine. The experiments employed rabbit polyclonal antibodies against the major Na transporters of the proximal tubule (the type 3 Na-H exchanger [NHE3]), the thick ascending limb of Henle's loop (the bumetanide-sensitive Na-K-2Cl cotransporter [NKCC2]), and the distal convoluted tubule (the thiazide-sensitive Na-CI cotransporter [NCC]) in immunoblotting experiments. All three of these transporters were readily detectable as high molecular weight complexes present in low-density membrane fractions from urine of normal rats. Cross linking studies of NHE3, NKCC2, and NCC revealed that high molecular weight complexes are normally present in renal tissue. The molecular weights of the complexes in urine matched those of the cross-linked complexes in native kidney tissue. The presence in urine of integral membrane proteins representative of each nephron segment raises the possibility that limited or comprehensive proteomic analysis of urine samples may be useful in clinical settings. C1 NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Dept Physiol, Bethesda, MD 20814 USA. Whitehead Inst Biomed Engn, Cambridge, MA USA. Howard Hughes Med Inst, Cambridge, MA USA. RP Knepper, MA (reprint author), NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bldg 10,Room 6N260,10 Ctr Dr MSC 1603, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z01 HL001285-21, Z99 HL999999]; NHLBI NIH HHS [Z01-HL-01282-KE] NR 20 TC 60 Z9 61 U1 1 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 NOV PY 2000 VL 11 IS 11 BP 2128 EP 2132 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 368VQ UT WOS:000090138500021 PM 11053490 ER PT J AU Flood, A Schatzkin, A AF Flood, A Schatzkin, A TI Colorectal cancer: Does it matter if you eat your fruits and vegetables? SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Editorial Material ID COLON-CANCER; RISK; MEN C1 NCI, Nutrit Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Schatzkin, A (reprint author), NIH, Execut Plaza N,Rm 211,6130 Execut Blvd, Rockville, MD 20852 USA. NR 16 TC 20 Z9 20 U1 0 U2 2 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD NOV 1 PY 2000 VL 92 IS 21 BP 1706 EP 1707 DI 10.1093/jnci/92.21.1706 PG 2 WC Oncology SC Oncology GA 368HC UT WOS:000090110800002 PM 11058607 ER PT J AU Mark, SD Qiao, YL Dawsey, SM Wu, YP Katki, H Gunter, EW Fraumeni, JF Blot, WJ Dong, ZW Taylor, PR AF Mark, SD Qiao, YL Dawsey, SM Wu, YP Katki, H Gunter, EW Fraumeni, JF Blot, WJ Dong, ZW Taylor, PR TI Prospective study of serum selenium levels and incident esophageal and gastric cancers SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID GLUTATHIONE-PEROXIDASE ACTIVITY; NUTRITION INTERVENTION TRIALS; DISEASE-SPECIFIC MORTALITY; SUBSEQUENT RISK; LUNG-CANCER; VITAMIN-E; FINNISH MEN; GASTROINTESTINAL CANCER; MINERAL SUPPLEMENTATION; SEROLOGIC PRECURSORS AB Background: From March 1986 through May 1991, we conducted a randomized nutritional intervention trial, the General Population Trial, in Linxian, China, a region with epidemic rates of squamous esophageal and adenomatous gastric cardia cancers, We found that participants who received selenium, beta -carotene, and vitamin E had significantly lower cancer mortality rates than those who did not. In the current study, we examined the relationship between selenium levels measured in pretrial (1985) sera from participants and the subsequent risk of developing squamous esophageal, gastric cardia, and gastric non-cardia cancers during the trial. Methods: This study was designed and analyzed in accord with a stratified case-cohort sampling scheme, with the six strata defined by sex and three age categories. We measured serum selenium levels in 590 case subjects with esophageal cancer, 402 with gastric cardia cancers, and 87 with gastric non-cardia cancers as well as in 1062 control subjects. Relative risks (RRs), absolute risks, and population attributable risk for cancers were estimated on the basis of the Cox proportional hazards models. All statistical tests are two-sided. Results: We found highly significant inverse associations of serum selenium levels with the incidence of esophageal (P for trend <10(-4)) and gastric cardia (P for trend <10(-6)) cancers. The RR and 95% confidence interval (CI) for comparison of highest to lowest quartile of serum selenium was 0.56 (95% CI = 0.44-0.71) for esophageal cancer and 0.47 (95% CI = 0.33-0.65) for gastric cardia cancer. The population proportion of these cancers that is attributable to low selenium levels was 26.4% (95% CI = 14.45-38.36), We found no evidence for a gradient of serum selenium associated with incidence of gastric non-cardia cancer (P for trend = .96), with an RR of 1.07 (95% CI = 0.55-2.08) for the highest to lowest quartile of serum selenium, Conclusions: Our study supports findings from previous prospective studies and randomized trials that variations in selenium levels affect the incidence of certain cancers. In the United States, where intervention trials of selenium are in the planning stages, consideration should be given to including populations at high risk for squamous esophageal and gastric cardia cancers. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NCI, Div Clin Sci, Bethesda, MD 20892 USA. Chinese Acad Med Sci, Inst Canc, Dept Epidemiol, Beijing 100021, Peoples R China. Ctr Dis Control & Prevent, Natl Hlth & Nutr Examinat Survey Lab, Atlanta, GA USA. Int Epidemiol Inst, Rockville, MD USA. RP Mark, SD (reprint author), NIH, 6120 Execut Blvd,Rm 8036,MSC 7244, Rockville, MD 20852 USA. RI Qiao, You-Lin/B-4139-2012; Katki, Hormuzd/B-4003-2015 OI Qiao, You-Lin/0000-0001-6380-0871; NR 62 TC 161 Z9 171 U1 0 U2 5 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD NOV 1 PY 2000 VL 92 IS 21 BP 1753 EP 1763 DI 10.1093/jnci/92.21.1753 PG 11 WC Oncology SC Oncology GA 368HC UT WOS:000090110800012 PM 11058618 ER PT J AU French, JE Saulnier, M AF French, JE Saulnier, M TI Benzene leukemogenesis: An environmental carcinogen-induced tissue-specific model of neoplasia using genetically altered mouse models SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article; Proceedings Paper CT Benzene State-of-the-Science Workshop CY DEC 16-17, 1998 CL UNIV OTTAWA, OTTAWA, CANADA SP Amer Petr Inst, Canadian Petr Prod Inst, NIEHS, US EPA, Western States Petr Assoc HO UNIV OTTAWA ID MICE; EXPOSURE; LEUKEMIA; GENE C1 NIEHS, NIH, Res Triangle Pk, NC 27709 USA. N Carolina State Univ, Raleigh, NC 27695 USA. RP French, JE (reprint author), NIEHS, NIH, POB 12233,MD F1-05, Res Triangle Pk, NC 27709 USA. NR 16 TC 7 Z9 8 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD NOV PY 2000 VL 61 IS 5-6 BP 377 EP 379 DI 10.1080/00984100050166389 PG 3 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 369VA UT WOS:000165086700008 PM 11086942 ER PT J AU Hayes, RB Yin, SN Rothman, N Dosemeci, M Li, GL Travis, LT Smith, MT Linet, MS AF Hayes, RB Yin, SN Rothman, N Dosemeci, M Li, GL Travis, LT Smith, MT Linet, MS TI Benzene and lymphohematopoietic malignancies in China SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article; Proceedings Paper CT Benzene State-of-the-Science Workshop CY DEC 16-17, 1998 CL UNIV OTTAWA, OTTAWA, CANADA SP Amer Petr Inst, Canadian Petr Prod Inst, NIEHS, US EPA, Western States Petr Assoc HO UNIV OTTAWA ID ACUTE MYELOID-LEUKEMIA; EXPOSED WORKERS; OCCUPATIONAL EXPOSURE; HEMATOPOIETIC MALIGNANCIES; RETROSPECTIVE COHORT; MULTIPLE-MYELOMA; PLIOFILM COHORT; RISK; MORTALITY; DISORDERS C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Chinese Acad Prevent Med, Beijing, Peoples R China. Univ Calif Berkeley, Sch Publ Hlth, Div Environm Hlth Sci, Berkeley, CA 94720 USA. RP Hayes, RB (reprint author), NCI, Div Canc Epidemiol & Genet, EPS 8114, Bethesda, MD 20892 USA. NR 39 TC 31 Z9 33 U1 0 U2 2 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD NOV PY 2000 VL 61 IS 5-6 BP 419 EP 432 DI 10.1080/00984100050166442 PG 14 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 369VA UT WOS:000165086700014 PM 11086948 ER PT J AU Smith, MT Rothman, N AF Smith, MT Rothman, N TI Biomarkers in the molecular epidemiology of benzene-exposed workers SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article; Proceedings Paper CT Benzene State-of-the-Science Workshop CY DEC 16-17, 1998 CL UNIV OTTAWA, OTTAWA, CANADA SP Amer Petr Inst, Canadian Petr Prod Inst, NIEHS, US EPA, Western States Petr Assoc HO UNIV OTTAWA ID IN-SITU HYBRIDIZATION; LONG ARM DELETION; HUMAN-LYMPHOCYTES; DNA PROBES; LEUKEMIA; MUTATION; RISK; HYDROQUINONE; CHROMOSOME-5; ABERRATIONS C1 Univ Calif Berkeley, Sch Publ Hlth, Div Environm Hlth Sci, Berkeley, CA 94720 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Smith, MT (reprint author), Univ Calif Berkeley, Sch Publ Hlth, Div Environm Hlth Sci, 216 Warren Hall, Berkeley, CA 94720 USA. FU NIEHS NIH HHS [P30 ES01896, P42 ES04705, R01 ES06721] NR 25 TC 18 Z9 19 U1 0 U2 1 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD NOV PY 2000 VL 61 IS 5-6 BP 439 EP 445 DI 10.1080/00984100050166460 PG 7 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 369VA UT WOS:000165086700016 PM 11086950 ER PT J AU Bayliss, D Sonawane, B AF Bayliss, D Sonawane, B TI Issues for discussion: Benzene-induced leukemia-human studies SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article; Proceedings Paper CT Benzene State-of-the-Science Workshop CY DEC 16-17, 1998 CL UNIV OTTAWA, OTTAWA, CANADA SP Amer Petr Inst, Canadian Petr Prod Inst, NIEHS, US EPA, Western States Petr Assoc HO UNIV OTTAWA ID RETROSPECTIVE COHORT; EXPOSED WORKERS; CHINA; MORTALITY C1 NCI, Natl Ctr Environm Assessment, US EPA, Chinese Epidemiol Study, Bethesda, MD 20892 USA. RP Bayliss, D (reprint author), NCI, Natl Ctr Environm Assessment, US EPA, Chinese Epidemiol Study, Bethesda, MD 20892 USA. NR 12 TC 1 Z9 1 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD NOV PY 2000 VL 61 IS 5-6 BP 467 EP 470 DI 10.1080/00984100050166497 PG 4 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 369VA UT WOS:000165086700019 PM 11086953 ER PT J AU Portier, CJ Parham, FM AF Portier, CJ Parham, FM TI Comments on a biochemical model of cyclophosphamide hematotoxicity SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article; Proceedings Paper CT Hematotoxicity Modeling Workshop CY DEC 15, 1998 CL UNIV OTTAWA, OTTAWA, CANADA SP Amer Petr Inst HO UNIV OTTAWA C1 NIEHS, Lab Computat Biol & Risk Anal, Res Triangle Pk, NC 27709 USA. RP Portier, CJ (reprint author), NIEHS, Lab Computat Biol & Risk Anal, POB 12233, Res Triangle Pk, NC 27709 USA. RI Portier, Christopher/A-3160-2010 OI Portier, Christopher/0000-0002-0954-0279 NR 8 TC 0 Z9 0 U1 1 U2 1 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD NOV PY 2000 VL 61 IS 5-6 BP 525 EP 528 DI 10.1080/00984100050166596 PG 4 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 369VA UT WOS:000165086700029 PM 11086963 ER PT J AU Leach, FS Velasco, A Hsieh, JT Sagalowsky, AI McConnell, JD AF Leach, FS Velasco, A Hsieh, JT Sagalowsky, AI McConnell, JD TI The mismatch repair gene hMSH2 is mutated in the prostate cancer cell line LNCaP SO JOURNAL OF UROLOGY LA English DT Article DE base pair mismatch; DNA repair; microsatellite repeats; prostatic neoplasms ID NONPOLYPOSIS COLORECTAL-CANCER; MICROSATELLITE INSTABILITY; SOMATIC MUTATIONS; EXPRESSION; SEQUENCES; HOMOLOG; COLON; MSH2 AB Purpose: Mismatch repair genes are responsible for the coordinated correction of misincorporated nucleotides formed during DNA replication. Inactivating and inherited mutations in the prototypic mismatch repair gene hMSH2 have been described in a cancer predisposition syndrome known as hereditary nonpolyposis colon cancer. Patients with hereditary nonpolyposis colon cancer are at increased risk for colon cancer and extracolonic cancers such as upper tract transitional cell carcinoma but not prostate cancer. We investigated expression of hMSH2 in prostate cancer cell lines using genetic and molecular analysis. Materials and Methods: We used the 3 well described prostate cancer cell lines, DU145, LNCaP and PC3. Western blot analysis with monoclonal antibody to hMSH2 was used to assess expression. Southern blot and polymerase chain reaction of genomic DNA were used to identify genetic alterations in the hMSH2 gene. Single cell cloning, dinucleotide repeats and BAT-26 were used to assess the cell lines for microsatellite instability. Results: The prostate cancer cell line LNCaP did not express hMSH2 and was found to have a homozygous deletion of hMSH2 exons 9 to 16, resulting in truncation of the protein. While microsatellite analysis did not reveal alterations at the BAT-26 locus, single cell cloning produced several LNCaP subclones with alteration at 1 dinucleotide repeat. Conclusions: The well described prostate cancer cell line LNCaP has a mutation in the hMSH2 gene, resulting in loss of expression and possible evidence of microsatellite instability. To our knowledge our finding is the first demonstration of a genetic alteration in hMSH2 in a prostate cancer cell line. C1 NCI, Urol Oncol Branch, NIH, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Urol, Dallas, TX USA. RP Leach, FS (reprint author), NCI, Urol Oncol Branch, NIH, 10 Ctr Dr,Bldg 10 Room 2B47, Bethesda, MD 20892 USA. FU NCI NIH HHS [T32-CA66187-01A1] NR 24 TC 34 Z9 34 U1 0 U2 2 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 NOV PY 2000 VL 164 IS 5 BP 1830 EP 1833 DI 10.1016/S0022-5347(05)67115-7 PG 4 WC Urology & Nephrology SC Urology & Nephrology GA 363DT UT WOS:000089820300114 PM 11025778 ER PT J AU Pinto, VB Prasad, S Yewdell, J Bennink, J Hughes, SH AF Pinto, VB Prasad, S Yewdell, J Bennink, J Hughes, SH TI Restricting expression prolongs expression of foreign genes introduced into animals by retroviruses SO JOURNAL OF VIROLOGY LA English DT Article ID RECOMBINANT ADENOVIRUSES; IMMUNE-RESPONSES; FACTOR-VIII; IN-VIVO; TRANSGENIC MICE; DNA-SYNTHESIS; VIRAL GENES; FACTOR-IX; T-CELLS; VECTORS AB If foreign genes are ubiquitously expressed in mice using a viral vector, expression is abrogated by CD8(+) cells in 2 to 4 weeks. However, if the expression of the genes is confined to skeletal muscle cells, the CD8(+) T-cell response is much weaker and expression is maintained for more than 6 weeks. These data show that restricting the expression of foreign genes to skeletal muscle cells and presumably to other cells that are inefficient at antigen presentation can prolong the expression of a foreign gene product. C1 NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Frederick, MD 21702 USA. NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Hughes, SH (reprint author), NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, POB B,Bldg 539, Frederick, MD 21702 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012 NR 38 TC 14 Z9 15 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD NOV PY 2000 VL 74 IS 21 BP 10202 EP 10206 DI 10.1128/JVI.74.21.10202-10206.2000 PG 5 WC Virology SC Virology GA 363DN UT WOS:000089819900043 PM 11024149 ER PT J AU Halvas, EK Svarovskaia, ES Pathak, VK AF Halvas, EK Svarovskaia, ES Pathak, VK TI Role of murine leukemia virus reverse transcriptase deoxyribonucleoside triphosphate-binding site in retroviral replication and in vivo fidelity SO JOURNAL OF VIROLOGY LA English DT Article ID DNA-SYNTHESIS; CATALYTIC MECHANISM; ANGSTROM RESOLUTION; MUTATIONAL ANALYSIS; GENETIC-VARIATION; HIV-1; RESISTANCE; MUTAGENESIS; COMBINATION; THERAPY AB Retroviral populations exhibit a high evolutionary potential, giving rise to extensive genetic variation. Error-prone DNA synthesis catalyzed by reverse transcriptase (RT) generates variation in retroviral populations. Structural features within RTs are likely to contribute to the high rate of errors that occur during reverse transcription. We sought to determine whether amino acids within murine leukemia virus (MLV) RT that contact the deoxyribonucleoside triphosphate (dNTP) substrate are important for in vivo fidelity of reverse transcription. We utilized the previously described ANGIE P encapsidating cell line, which expresses the amphotropic MLV envelope and a retroviral vector (pGA-1). pGA-1 expresses the bacterial P-galactosidase gene (lacZ), which serves as a reporter of mutations. Extensive mutagenesis was performed on residues likely to interact with the dNTP substrate, and the effects of these mutations on the fidelity of reverse transcription were determined. As expected, most substitution mutations of amino acids that directly interact with the cNTP substrate significantly reduced viral titers (>10,000-fold), indicating that these residues played a critical role in catalysis and viral replication. However, the D153A and A154S substitutions, which are predicted to affect the interactions with the triphosphate, resulted in statistically significant increases in the mutation rate. In addition, the conservative substitution F155W, which may affect interactions with the base and the ribose, increased the mutation rate 2.8-fold. Substitutions of residues in the vicinity of the dNTP-binding site also resulted in statistically significant decreases in fidelity (1.3- to 2.4-fold). These results suggest that mutations of residues that contact the substrate dNTP can affect viral replication as well as alter the fidelity of reverse transcription. C1 NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Frederick, MD 21702 USA. W Virginia Univ, Mary Babb Randolph Canc Ctr, Morgantown, WV 26506 USA. W Virginia Univ, Dept Biochem, Morgantown, WV 26506 USA. RP Pathak, VK (reprint author), NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Bldg 535,Rm 334, Frederick, MD 21702 USA. FU NCI NIH HHS [CA58875] NR 50 TC 17 Z9 17 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 NOV PY 2000 VL 74 IS 22 BP 10349 EP 10358 DI 10.1128/JVI.74.22.10349-10358.2000 PG 10 WC Virology SC Virology GA 367WE UT WOS:000090083800010 PM 11044079 ER PT J AU Silverstein, PS Mackay, GA Mukherjee, S Li, Z Piatak, M Lifson, JD Narayan, O Kumar, A AF Silverstein, PS Mackay, GA Mukherjee, S Li, Z Piatak, M Lifson, JD Narayan, O Kumar, A TI Pathogenic simian/human immunodeficiency virus SHIVKU inoculated into immunized macaques caused infection, but virus burdens progressively declined with time SO JOURNAL OF VIROLOGY LA English DT Article ID PIG-TAILED MACAQUES; CD4(+) T-CELLS; BETA-CHEMOKINES; RHESUS-MONKEYS; AIDS; VACCINE; PROTECTION; LIVE; LYMPHOCYTES; INDUCTION AB Using the simian immunodeficiency virus/human immunodeficiency virus (SHIV)-macaque model of AIDS, we had shown in a previous report that a live, nonpathogenic strain of SHIV,further attenuated by deletion of the vpu gene and inoculated orally into adult macaques, had effectively prevented AIDS following vaginal inoculation with pathogenic SHIVKU. Examination of lymph nodes from the animals at 18 weeks postchallenge had shown that all six animals were persistently infected with challenge virus. We report here on a 2-year follow-up study on the nature of the persistent infections in these animals. DNA of the vaccine virus was present in the lymph nodes at all time points tested, as far as 135 weeks postchallenge. In contrast, the DNA of SHIVKU became undetectable in one animal by week 55 and in three others by week 63. These four macaques have remained negative for SHIVKU DNA as far as the last time point examined at week 135. Quantification of the total viral DNA concentration in lymph nodes during the observation period showed a steady decline. All animals developed neutralizing antibody and cytotoxic-T-lymphocyte responses to SHIVKU that persisted throughout the observation period. Vaccine-like viruses were isolated from two animals, and a SHIVKU-like virus was isolated from one of the two macaques that remained positive for SHIVKU DNA. There was no evidence of recombination between the vaccine and the challenge viruses. Thus, immunization with the live vaccine not only prevented disease but also contributed to the steady decline in the virus burdens in the animals. C1 Univ Kansas, Med Ctr, Dept Microbiol Mol Genet & Immunol, Marion Merrell Dow Lab Viral Pathogenesis, Kansas City, KS 66160 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, AIDS Vaccine Program, Frederick, MD 21702 USA. RP Kumar, A (reprint author), Univ Kansas, Med Ctr, Dept Microbiol Mol Genet & Immunol, Marion Merrell Dow Lab Viral Pathogenesis, 3901 Rainbow Blvd, Kansas City, KS 66160 USA. FU NCRR NIH HHS [R01 RR006753, RR-13152, RR-06753]; NIAID NIH HHS [AI-29382] NR 31 TC 21 Z9 21 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 NOV PY 2000 VL 74 IS 22 BP 10489 EP 10497 DI 10.1128/JVI.74.22.10489-10497.2000 PG 9 WC Virology SC Virology GA 367WE UT WOS:000090083800024 PM 11044093 ER PT J AU Hallak, LK Spillmann, D Collins, PL Peeples, ME AF Hallak, LK Spillmann, D Collins, PL Peeples, ME TI Glycosaminoglycan sulfation requirements for respiratory syncytial virus infection SO JOURNAL OF VIROLOGY LA English DT Article ID ATTACHMENT GLYCOPROTEIN-G; SURFACE HEPARAN-SULFATE; HERPES-SIMPLEX VIRUS; SH GENE; NUCLEOTIDE-SEQUENCE; BINDING DOMAIN; A27L PROTEIN; CELL-FUSION; SUBGROUP-B; IN-VITRO AB Glycosaminoglycans (GAGs) on the surface of cultured cells are important in the first step of efficient respiratory syncytial virus (RSV) infection. We evaluated the importance of sulfation, the major biosynthetic modification of GAGs, using an improved recombinant green fluorescent protein-expressing RSV (rgRSV) to assay infection. Pretreatment of HEp-2 cells with 50 mM sodium chlorate, a selective inhibitor of sulfation, for 48 h prior to inoculation reduced the efficiency of rgRSV infection to 40%. Infection of a CHO mutant cell line deficient in N-sulfation was three times less efficient than infection of the parental CHO cell line, indicating that N-sulfation is important. In contrast, infection of a cell line deficient in 2-O-sulfation was as efficient as infection of the parental cell line, indicating that 2-O-sulfation is not required for RSV infection. Incubating RSV with the purified soluble heparin, the prototype GAG, before inoculation had previously been shown to neutralize its infectivity. Here we tested chemically modified heparin chains that lack their N-, C6-O-, or C2-O-sulfate groups. Only heparin chains lacking the N-sulfate group lost the ability to neutralize infection, confirming that N-sulfation, but not C6-O- or C2-O-sulfation, is important for RSV infection. Analysis of heparin fragments identified the 10-saccharide chain as the minimum size that can neutralize RSV infectivity. Taken together, these results show that, while sulfate modification is important for the ability of GAGs to mediate RSV infection, only certain sulfate groups are required. This specificity indicates that the role of cell surface GAGs in RSV infection is not based on a simple charge interaction between the virus and sulfate groups but instead involves a specific GAG structural configuration that includes N-sulfate and a minimum of 10 saccharide subunits. These elements, in addition to iduronic acid demonstrated previously (L. K. Hallak, P. L. Collins, W Knudson, and M. E. Peeples, Virology 271:264-275, 2000), partially define cell surface molecules important for RSV infection of cultured cells. C1 Rush Presbyterian St Lukes Med Ctr, Dept Immunol Microbiol, Chicago, IL 60612 USA. Biomed Ctr, Dept Med Biochem & Microbiol, Sect Med Biochem, SE-75123 Uppsala, Sweden. NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Peeples, ME (reprint author), Rush Presbyterian St Lukes Med Ctr, Dept Immunol Microbiol, W Congress Pkwy, Chicago, IL 60612 USA. NR 43 TC 157 Z9 163 U1 0 U2 6 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 NOV PY 2000 VL 74 IS 22 BP 10508 EP 10513 DI 10.1128/JVI.74.22.10508-10513.2000 PG 6 WC Virology SC Virology GA 367WE UT WOS:000090083800026 PM 11044095 ER PT J AU Wang, SW Kozlowski, PA Schmelz, G Manson, K Wyand, MS Glickman, R Montefiori, D Lifson, JD Johnson, RP Neutra, MR Aldovini, A AF Wang, SW Kozlowski, PA Schmelz, G Manson, K Wyand, MS Glickman, R Montefiori, D Lifson, JD Johnson, RP Neutra, MR Aldovini, A TI Effective induction of simian immunodeficiency virus-specific systemic and mucosal immune responses in primates by vaccination with proviral DNA producing intact but noninfectious virions SO JOURNAL OF VIROLOGY LA English DT Article ID LYMPH-NODE IMMUNIZATION; CYTOTOXIC T-LYMPHOCYTES; FEMALE RHESUS MACAQUES; NONHUMAN-PRIMATES; REVERSE-TRANSCRIPTASE; ANTIBODY-RESPONSES; MONONUCLEAR-CELLS; VAGINAL CHALLENGE; GENITAL-TRACT; SEX WORKERS AB We report a pilot evaluation of a DNA vaccine producing genetically inactivated simian immunodeficiency virus (SIV) particles in primates, with a focus on eliciting mucosal immunity. Our results demonstrate that DNA vaccines can be used to stimulate strong virus-specific mucosal immune responses in primates. The levels of immunoglobulin A (IgA) detected in rectal secretions of macaques that received the DNA vaccine intradermally and at the rectal mucosa were the most striking of all measured immune responses and were higher than usually achieved through natural infection. However, cytotoxic T lymphocyte responses were generally low and sporadically present in different animals. Upon rectal challenge with cloned SIVmac239, resistance to infection was observed, but some animals with high SIV-specific IgA levels in rectal secretions became infected. Our results suggest that high levels of IgA alone are not sufficient to prevent the establishment of chronic infection, although mucosal IgA responses may have a role in reducing the infectivity of the initial viral inoculum. C1 Childrens Hosp, Dept Med, Boston, MA 02115 USA. Harvard Univ, Sch Med, Dept Pediat, Boston, MA 02115 USA. Primedica, Worcester, MA USA. Duke Univ, Durham, NC USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, AIDS Vaccine Program,Retroviral Pathogenesis Lab, Frederick, MD USA. Harvard Univ, New England Reg Primate Res Ctr, Sch Med, Southborough, MA 01772 USA. Massachusetts Gen Hosp, Partners AIDS Res Ctr, Charlestown, MA USA. Massachusetts Gen Hosp, Infect Dis Unit, Charlestown, MA USA. RP Aldovini, A (reprint author), Childrens Hosp, Dept Med, Enders 609,300 Longwood Ave, Boston, MA 02115 USA. RI Wang, Shainn-Wei/B-9644-2009 FU NCRR NIH HHS [K26 RR000168, P51 RR000168, RR00168]; NIAID NIH HHS [AI34757, R01 AI041365, R56 AI041365, AI41365] NR 54 TC 52 Z9 56 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 NOV PY 2000 VL 74 IS 22 BP 10514 EP 10522 DI 10.1128/JVI.74.22.10514-10522.2000 PG 9 WC Virology SC Virology GA 367WE UT WOS:000090083800027 PM 11044096 ER PT J AU Reiser, J Lai, ZN Zhang, XY Brady, RO AF Reiser, J Lai, ZN Zhang, XY Brady, RO TI Development of multigene and regulated lentivirus vectors SO JOURNAL OF VIROLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; VIVO GENE DELIVERY; NONDIVIDING CELLS; HIV-1 VECTOR; IN-VIVO; EFFICIENT TRANSDUCTION; RETROVIRAL VECTORS; CD34(+) CELLS; EXPRESSION; TYPE-1 AB Previously we described safe and efficient three-component human immunodeficiency virus type I (HIV-1)-based gene transfer systems for delivery of genes into nondividing cells (H. Mochizuki, J. P. Schwartz, K. Tanaka, R. O. Brady, and J. Reiser, J. Virol. 72:8873-8883, 1998). To apply such vectors in anti-HIV gene therapy strategies and to express multiple proteins in single target cells, we have engineered HIV-1 vectors for the concurrent expression of multiple transgenes. Single-gene vectors, bicistronic vectors, and multigene vectors expressing up to three exogenous genes under the control of two or three different transcriptional units, placed within the viral gag-pol coding region and/or the viral nef and env genes, were designed. The genes encoding the enhanced version of green fluorescent protein (EGFP), mouse heat-stable antigen (HSA), and bacterial neomycin phosphotransferase were used as models whose expression was detected by fluorescence-activated cell sorting, fluorescence microscopy, and G418 selection. Coexpression of these reporter genes in contact-inhibited primary human skin fibroblasts (HSFs) persisted for at least 6 weeks in culture. Coexpression of the HSA and EGFP reporter genes was also achieved following cotransduction of target cells using two separate lentivirus vectors encoding HSA and EGFP, respectively. For the regulated expression of transgenes, tetracycline (Tet)-regulatable lentivirus vectors encoding the reverse Tet transactivator (rtTA) and EGFP controlled by a Tet-responsive element (TRE) were constructed. A binary HIV-l-based vector system consisting of a lentivirus encoding rtTA and a second lentivirus harboring a TRE driving the EGFP reporter gene was also designed. Doxycycline-modulated expression of the EGFP transgene was confirmed in transduced primary HSFs. These versatile vectors can potentially be used in a wide range of gene therapy applications. C1 Louisiana State Univ, Sch Med, Dept Med, New Orleans, LA 70112 USA. Louisiana State Univ, Sch Med, Dept Microbiol Immunol & Parasitol, New Orleans, LA 70112 USA. Louisiana State Univ, Sch Med, Gene Therapy Program, New Orleans, LA 70112 USA. NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Reiser, J (reprint author), Louisiana State Univ, Hlth Sci Ctr, LSU Gene Therapy Program, MEB 3205,1901 Perdido St, New Orleans, LA 70112 USA. NR 69 TC 87 Z9 92 U1 2 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD NOV PY 2000 VL 74 IS 22 BP 10589 EP 10599 DI 10.1128/JVI.74.22.10589-10599.2000 PG 11 WC Virology SC Virology GA 367WE UT WOS:000090083800034 PM 11044103 ER PT J AU Zheng, ZM Reid, ES Baker, CC AF Zheng, ZM Reid, ES Baker, CC TI Utilization of the bovine papillomavirus type 1 late-stage-specific nucleotide 3605 3 ' splice site is modulated by a novel exonic bipartite regulator but not by an intronic purine-rich element SO JOURNAL OF VIROLOGY LA English DT Article ID PRE-MESSENGER-RNA; IMMUNODEFICIENCY-VIRUS TYPE-1; FIBRONECTIN EDA EXON; SR-PROTEINS; ALTERNATIVE EXON; BRANCH POINT; IN-VIVO; FACTOR U2AF(35); TAT EXON-2; U2 SNRNA AB Bovine papillomavirus type 1 (BPV-1) late gene expression is regulated at both transcriptional and posttranscriptional levels. Maturation of the capsid protein (L1) pre-mRNA requires a switch in 3' splice site utilization. This switch involves activation of the nucleotide (nt) 3605 3' splice site, which is utilized only in fully differentiated keratinocytes during late stages of the virus life cycle, Our previous studies of the mechanisms that regulate BPV-1 alternative splicing identified three cis-acting elements between these two splice sites. Two purine-rich exonic splicing enhancers, SE1 and SE2, are essential for preferential utilization of the nt 3225 3' splice site at early stages of the virus life cycle. Another cis-acting element, exonic splicing suppressor 1 (ESS1), represses use of the nt 3225 3' splice site, In the present study, we investigated the late-stage-specific nt 3605 3' splice site and showed that it has suboptimal features characterized by a nonconsensus branch point sequence and a weak polypyrimidine track with interspersed purines. In vitro and in vivo experiments showed that utilization of the nt 3605 3' splice site was not affected by SE2, which is intronically located with respect to the nt 3605 3' splice site. The intronic location and sequence composition of SE2 are similar to those of the adenovirus IIIa repressor element, which has been shown to inhibit use of a downstream 3' splice site. Further studies demonstrated that the nt 3605 3' splice site is controlled by a novel exonic bipartite element consisting of an AC-rich exonic splicing enhancer (SE4) and an exonic splicing suppressor (ESS2) with a UGGU motif. Functionally, this newly identified bipartite element resembles the bipartite element composed of SE1 and ESS1, SE4 also functions on a heterologous 3' splice site. In contrast, ESS2 functions as an exonic splicing suppressor only in a 3'-splice-site-specific and enhancer-specific manner. Our data indicate that BPV-1 splicing regulation is very complex and is likely to be controlled by multiple splicing factors during keratinocyte differentiation. C1 NCI, Div Basic Sci, Basic Res Lab, NIH, Bethesda, MD 20892 USA. RP Zheng, ZM (reprint author), NCI, DCS, HAMB, Bldg 10,Room 13N240,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 61 TC 25 Z9 25 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD NOV PY 2000 VL 74 IS 22 BP 10612 EP 10622 DI 10.1128/JVI.74.22.10612-10622.2000 PG 11 WC Virology SC Virology GA 367WE UT WOS:000090083800036 PM 11044105 ER PT J AU Feng, YX Moore, SP Garfinkel, DJ Rein, A AF Feng, YX Moore, SP Garfinkel, DJ Rein, A TI The genomic RNA in Ty1 virus-like particles is dimeric SO JOURNAL OF VIROLOGY LA English DT Article ID YEAST RETROTRANSPOSON TY1; ACID-CHAPERONE ACTIVITY; PRIMER-BINDING-SITE; SACCHAROMYCES-CEREVISIAE; ELEMENT TRANSPOSITION; EXTENDED DUPLEX; RECOMBINATION; DIMERIZATION; PROTEASE; MATURATION AB The yeast retrotransposon Ty1 resembles retroviruses in a number of important respects but also shows several fundamental differences from them. We now report that, as in retroviruses, the genomic RNA in Ty1 virus-like particles is dimeric. The Ty1 dimers also resemble retroviral dimers in that they are stabilized during the proteolytic maturation of the particle. The statbilization of the dimer suggests that one of the cleavage products of TyA1 possesses nucleic acid chaperone activity. C1 NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Gene Regulat & Chromosome Biol Lab, Frederick, MD 21702 USA. RP Rein, A (reprint author), NCI, Frederick Canc Res & Dev Ctr, HIV Drug Resistance Program, Bldg 535,Room 211, Frederick, MD 21702 USA. NR 24 TC 29 Z9 31 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 NOV PY 2000 VL 74 IS 22 BP 10819 EP 10821 DI 10.1128/JVI.74.22.10819-10821.2000 PG 3 WC Virology SC Virology GA 367WE UT WOS:000090083800061 PM 11044130 ER PT J AU Ivey, FM Roth, SM Ferrell, RE Tracy, BL Lemmer, JT Hurlbut, DE Martel, GF Siegel, EL Fozard, JL Metter, EJ Fleg, JL Hurley, BF AF Ivey, FM Roth, SM Ferrell, RE Tracy, BL Lemmer, JT Hurlbut, DE Martel, GF Siegel, EL Fozard, JL Metter, EJ Fleg, JL Hurley, BF TI Effects of age, gender, and myostatin genotype on the hypertrophic response to heavy resistance strength training SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID SKELETAL-MUSCLE ADAPTATIONS; OLDER MEN; BODY-COMPOSITION; ELDERLY PEOPLE; RISK-FACTORS; WOMEN; MASS; GENE; ACTIVATION; CATTLE AB Background. Because of the scarcity of data available from direct comparisons of age and gender groups using the same relative training stimulus, it is unknown whether older individuals can increase their muscle mass as much as young individuals and whether women can increase as much as men in response to strength training (ST). In addition, little is known about whether the hypertrophic response to ST is affected by myostatin genotype, a candidate gene for muscle hypertrophy. Methods, Eleven young men (25 +/- 3 years, range 21-29 years), 11 young women (26 +/- 2 years, range 23-28 years), 12 older men (69 +/- 3 years, range 65-75 years), and 11 older women (68 +/- 2 years, range 65-73 pears) had bilateral quadriceps muscle volume measurements performed using magnetic resonance imaging (MRI) before and after ST and detraining. Training consisted of knee extension exercises of the dominant leg three times per week for 9 weeks. The contralateral limb was left untrained throughout the ST program. Following the unilateral training period, the subjects underwent 31 weeks of detraining during which no regular exercise was performed. Myostatin genotype was determined in a subgroup of 32 subjects, of which five female subjects were carriers of a myostatin gene variant. Results. A significantly greater absolute increase in muscle volume was observed in men than in women (204 +/- 20 vs 101 +/- 13 cm(3), p < .01), but there was no significant difference in muscle volume response to ST between young and older individuals. The gender effect remained after adjusting for baseline muscle volume. In addition, there was a significantly greater loss of absolute muscle volume after 31 weeks of detraining in men than in women (151 +/- 13 vs 88 +/- 7 cm(3), p < .05), but no significant difference between young and older individuals.;Myostatin genotype did not explain the hypertrophic response to ST when all 32 subjects were assessed. However, when only women were analyzed, those with the less common myostatin allele exhibited a 68% Larger increase in muscle volume in response to ST (p = .056). Conclusions. Aging does not affect the muscle mass response to either ST or detraining, whereas gender does, as men increased their muscle volume about twice as much in response to ST as did women and experienced larger losses in response to detraining than women. Young men were the only group that maintained muscle volume adaptation after 31 weeks of detraining. Although myostatin genotype may not explain the observed gender difference in the hypertrophic response to ST, a role for myostatin genotype may be indicated in this regard for women, but future studies are needed with larger subject numbers in each genotype group to confirm this observation. C1 Univ Maryland, Coll Hlth & Human Performance, Dept Kinesiol, College Pk, MD 20742 USA. Univ Maryland, Sch Med, Dept Gerontol, Baltimore, MD 21201 USA. Univ Pittsburgh, Dept Human Genet, Pittsburgh, PA USA. Univ Colorado, Dept Kinesiol & Appl Physiol, Boulder, CO 80309 USA. Tufts Univ, USDA, Human Nutr Res Ctr Aging, Boston, MA 02111 USA. Univ Maryland Eastern Shore, Dept Phys Therapy, Princess Anne, MD 21853 USA. Baltimore VA Med Ctr, Dept Radiol, Baltimore, MD USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. Morton Plant Mease Hlth Care, Florida Geriat Res Program, Clearwater, FL USA. RP Hurley, BF (reprint author), Univ Maryland, Coll Hlth & Human Performance, Dept Kinesiol, College Pk, MD 20742 USA. RI Fozard, James Leonard/B-3660-2009 FU NIA NIH HHS [AG-00268, AG42148]; PHS HHS [T3200219] NR 35 TC 122 Z9 128 U1 0 U2 11 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 NOV PY 2000 VL 55 IS 11 BP M641 EP M648 PG 8 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373WH UT WOS:000165312700011 PM 11078093 ER PT J AU Penninx, BWJH Ferrucci, L Leveille, SG Rantanen, T Pahor, M Guralnik, JM AF Penninx, BWJH Ferrucci, L Leveille, SG Rantanen, T Pahor, M Guralnik, JM TI Lower extremity performance in nondisabled older persons as a predictor of subsequent hospitalization SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID PHYSICAL PERFORMANCE; FUNCTIONAL DEPENDENCE; DISABILITY; EXERCISE; FRAILTY; ADULTS; RISK; MACARTHUR; STRENGTH AB Background. This study examines, in initially nondisabled older persons, the impact of reduced lower extremity performance on subsequent hospitalizations. Methods. A 4-year prospective cohort study was conducted among 3381 persons, aged 71 years and older, who initially reported no disability. At baseline, lower extremity performance was measured by an assessment of standing balance, a timed 2.4-m walk, and a timed test of rising from a chair five times. Data on subsequent hospital admissions and discharge diagnoses over 4 years were obtained from the Medicare database. Results. During the follow-up period, nondisabled persons with poor lower extremity performance spent significantly more days in the hospital (17.7 days) than those with intermediate and high performance (11.6 and 9.7 days, respectively). Poor lower extremity performance in nondisabled persons significantly predicted subsequent hospitalization over 4 years (relative risk for hospitalization in those with poor vs high performance: 1.78; 95% confidence interval, 1.45-2.17). This increased hospitalization risk could not be explained by several indicators of baseline health status. Increased hospitalization risks were especially found for geriatric conditions, such as dementia, detubitus ulcer, hip fractures, other fractures, pneumonia, dehydration, and acute infections. Conclusions. Even in persons who are currently nondisabled, a simple measure of lower extremity performance is predictive of subsequent hospitalization, especially for geriatric conditions. C1 Wake Forest Univ, Bowman Gray Sch Med, Sticht Ctr Aging, Winston Salem, NC 27157 USA. NIA, Epidemiol Demog & Biometry Program, NIH, Bethesda, MD 20892 USA. I Fraticini Natl Res Inst, INRCA, Dept Geriatr, Florence, Italy. Univ Jyvaskyla, Dept Hlth Sci, SF-40351 Jyvaskyla, Finland. RP Penninx, BWJH (reprint author), Wake Forest Univ, Bowman Gray Sch Med, Sticht Ctr Aging, Med Ctr Blvd, Winston Salem, NC 27157 USA. RI Rantanen, Taina/O-6579-2016 OI Rantanen, Taina/0000-0002-1604-1945 FU NIA NIH HHS [N01-AG-02106, N01-AG-02107, N01-AG-0215] NR 25 TC 193 Z9 195 U1 0 U2 6 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 NOV PY 2000 VL 55 IS 11 BP M691 EP M697 PG 7 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373WH UT WOS:000165312700018 PM 11078100 ER PT J AU Pieper, CF Rao, KMK Currie, MS Harris, TB Cohen, HJ AF Pieper, CF Rao, KMK Currie, MS Harris, TB Cohen, HJ TI Age, functional status, and racial differences in plasma D-dimer levels in community-dwelling elderly persons SO JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES LA English DT Article ID INCIDENT CARDIOVASCULAR-DISEASE; PLASMINOGEN-ACTIVATOR INHIBITOR; CORONARY-ARTERY DISEASE; DEGRADATION PRODUCT-D; MYOCARDIAL-INFARCTION; ENDOTHELIAL-CELLS; ACUTE STROKE; RISK-FACTORS; FIBRINOGEN; INTERLEUKIN-1 AB Background. Dysregulation of immunologic and coagulation systems is common in elderly persons and is associated with many diseases of aging. Thrombotic events are a major cause of morbidity and mortality in the elderly population. This study assesses whether D-dimer, a marker of fibrinolytic activity, varies systematically by demographic, health, and functional measures, and derives a prediction model for Factors related to D-dimer in a sample of community-dwelling elderly persons. Methods. D-dimer levels were assessed in a random sample of 1,727 community-dwelling elderly persons from five rural and urban counties in North Carolina in 1992, as part of the Established Populations for the Epidemiologic Studies of the Elderly (Duke University). All subjects were 72 years or older at the time of the blood draw. In addition, all subjects were surveyed yearly by telephone or in person each year from 1986 to 1992 for a variety of health, functional, and social Factors. Levels of D-dimer in 1992 were related cross-sectionally to demographics (age, race, education, income, gender, smoking), function (Nagi, Rosow-Breslau, Katz, Older Americans Resources and Services procedures instrumental activities of daily living), life satisfaction and self-rated health, self-reported diseases (heart attack, cancer, stroke, diabetes, and hypertension), and weight change from 1986 to 1992. Results. D-dimer levels increased with increasing age and functional disability. Among the health variables, only high blood pressure was predictive of D-dimer level. D-dimer levels were dramatically higher in blacks. Blacks were nearly four times more likely to have an extreme value of D-dimer (>600 mug/l) than whites when high D-dimer (yes/no) was analyzed, and blacks had an average level that was nearly 408 higher than whites in analyses of the continuous version of the outcome. This racial effect was not substantively affected in multivariable analyses with demographic and socioeconomic variables controlled. Race, age, functional status, current smoking, high blood pressure, and weight loss were related to level of D-dimer, and race, age, and functional status were related to the presence of a high D-dimer level (in the top 10% of the sample). Conclusions. Black, older, and functionally impaired persons had significantly higher levels of D-dimer in this sample of community-dwelling elderly persons. The findings for race were particularly striking and persisted even after controlling for smoking and other factors known to be related to thrombosis and were not mediated by social factors. This result may contribute to our understanding of the increased levels of thrombotic events found in these groups. C1 Duke Univ, Med Ctr, Ctr Aging & Human Dev, Durham, NC 27710 USA. Duke Univ, Med Ctr, Dept Community & Family Hlth, Div Biometr, Durham, NC 27710 USA. Duke Univ, Med Ctr, Claude D Pepper Older Amer Independence Ctr, Durham, NC 27710 USA. VA Med Ctr, Ctr Geriatr Res Educ & Clin, Durham, NC USA. Lewis Gale Clin, Salem, VA USA. NIA, NIH, Washington, DC USA. RP Pieper, CF (reprint author), Duke Univ, Med Ctr, Ctr Aging & Human Dev, Box 3003, Durham, NC 27710 USA. FU NIA NIH HHS [N01-AG1-2102, AG112618] NR 48 TC 63 Z9 66 U1 0 U2 3 PU GERONTOLOGICAL SOCIETY AMER PI WASHINGTON PA 1275 K STREET NW SUITE 350, WASHINGTON, DC 20005-4006 USA SN 1079-5006 J9 J GERONTOL A-BIOL JI J. Gerontol. Ser. A-Biol. Sci. Med. Sci. PD NOV PY 2000 VL 55 IS 11 BP M649 EP M657 PG 9 WC Geriatrics & Gerontology; Gerontology SC Geriatrics & Gerontology GA 373WH UT WOS:000165312700012 PM 11078094 ER PT J AU Allon, M Ornt, DB Schwab, SJ Rasmussen, C Delmez, JA Greene, T Kusek, JW Martin, AA Minda, S AF Allon, M Ornt, DB Schwab, SJ Rasmussen, C Delmez, JA Greene, T Kusek, JW Martin, AA Minda, S CA Hemodialysis HEMO Study Grp TI Factors associated with the prevalence of arteriovenous fistulas in hemodialysis patients in the HEMO Study SO KIDNEY INTERNATIONAL LA English DT Article DE angioaccess; age; race; diabetes; gender; vascular access; chronic renal failure ID VASCULAR ACCESS; POLYTETRAFLUOROETHYLENE; COMPLICATIONS; ANGIOACCESS; PREDICTORS; MORBIDITY; VEIN AB Background. Arteriovenous (AV) fistulas are the vascular access of choice for hemodialysis patients, but only about 20% of hemodialysis patients in the United States dialyze with fistulas. There is little information known about the factors associated with this low prevalence of fistulas. Methods. Multiple logistic regression analysis was used to evaluate the independent contribution of factors associated with AV fistula use among patients enrolled in the HEMO Study. The analysis was conducted in 1824 patients with fistulas or grafts at 45 dialysis units (15 clinical centers). Results. Thirty-four percent of the patients had fistulas. The prevalence of fistulas varied markedly from 4 to 77% among the individual dialysis units (P < 0.001). Multiple regression analysis revealed five demographic and clinical factors that were each independently associated with a lower likelihood of having a fistula, even after adjustment for dialysis unit. Specifically, the prevalence of fistulas was lower in females than males [adjusted odds ratio (AOR) 0.37, 95% CI, 0.28 to 0.48], lower in patients with peripheral vascular disease than in those without (AOR 0.55, 95% CI, 0.38 to 0.79), lower in blacks than in non-blacks (AOR 0.64, 95% CI, 0.46 to 0.89), lower in obese patients (AOR per 5 kg/m(2) body mass index, 0.76, 95% CI, 0.65 to 0.87), and lower in older patients (AOR per 10 years, 0.85, 95% CI, 0.78 to 0.94). The differences in the prevalence of fistulas among the dialysis units remained statistically significant (P < 0.001) after adjustment for these demographic and clinical factors. Finally, there were substantial variations in the prevalence of fistulas even among dialysis units in a single metropolitan area. Conclusions. Future efforts to increase the prevalence of fistulas in hemodialysis patients should be directed at both hemodialysis units and patient subpopulations with a low fistula prevalence. C1 Univ Alabama, Div Nephrol, Birmingham, AL 35294 USA. Univ Rochester, Rochester, NY USA. Duke Univ, Durham, NC 27706 USA. Cleveland Clin Fdn, Cleveland, OH 44195 USA. Washington Univ, St Louis, MO USA. NIDDKD, NIH, Bethesda, MD 20892 USA. New England Med Ctr, Boston, MA 02111 USA. RP Allon, M (reprint author), Univ Alabama, Div Nephrol, 1900 Univ Blvd,S,THT 647, Birmingham, AL 35294 USA. NR 23 TC 122 Z9 125 U1 0 U2 1 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD NOV PY 2000 VL 58 IS 5 BP 2178 EP 2185 DI 10.1046/j.1523-1755.2000.00391.x PG 8 WC Urology & Nephrology SC Urology & Nephrology GA 369CP UT WOS:000090154500037 PM 11044239 ER PT J AU Kopp, JB AF Kopp, JB TI BMP receptors in kidney SO KIDNEY INTERNATIONAL LA English DT Editorial Material DE bone morphogenetic protein; injury; TCF-beta gene super-family; acute renal failure ID BONE MORPHOGENETIC PROTEIN-7; MESENCHYME; EYE C1 NIDDK, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. RP Kopp, JB (reprint author), NIDDK, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 11 TC 14 Z9 17 U1 0 U2 0 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD NOV PY 2000 VL 58 IS 5 BP 2237 EP 2238 DI 10.1046/j.1523-1755.2000.00402.x PG 2 WC Urology & Nephrology SC Urology & Nephrology GA 369CP UT WOS:000090154500046 PM 11044248 ER PT J AU Bidus, KA Thomas, GR Ludlow, CL AF Bidus, KA Thomas, GR Ludlow, CL TI Effects of adductor muscle stimulation on speech in abductor spasmodic dysphonia SO LARYNGOSCOPE LA English DT Article; Proceedings Paper CT Biennial Speech Motor Control Conference CY FEB 03, 2000 CL SAN ANTONIO, TEXAS DE voice; muscle stimulation; spasmodic dysphonia ID LARYNGEAL NERVE STIMULATOR; BOTULINUM TOXIN; SPASTIC DYSPHONIA; ELECTRICAL-STIMULATION; ELECTRONIC CONTROL; DYSTONIA; INJECTIONS; CANINE AB Objective: To determine whether adductor laryngeal muscle stimulation might be a beneficial treatment alternative for abductor spasmodic dysphonia (ABSD), Study Design: Baseline comparisons were made on measures of voiceless consonant and syllable duration between patients with ABSD and normal control subjects, and speech and voice production with and without muscle stimulation were compared within 10 patients with ABSD, Methods: Baseline group comparisons were conducted on measures of syllable and voiceless consonant duration between the patients and the control subjects. Neuromuscular stimulation was applied to the thyroarytenoid or lateral cricoarytenoid muscles in the patients during extended phonation, and measures were made of fundamental frequency and sound pressure level in the stimulated and nonstimulated conditions. Voiceless consonant duration was compared with and without adductor laryngeal muscle stimulation during syllable repetitions and sentences in the patients, Results: Before stimulation, the patients had increased syllable durations in comparison with control subjects (P = .003), Repeated within-patient comparisons with and without stimulation demonstrated significant (P < .008) reductions in voiceless consonant durations during syllable repetition. The more severely affected patients had the greatest reductions in voiceless consonant duration during sentence production. Conclusions: Adductor muscle stimulation improved speech production in patients with ABSD, and the improvement was greatest in the most severely affected patients. Therefore adductor muscle stimulation has potential for benefiting patients with ABSD. C1 NINDS, Med Neurol Branch, Laryngeal & Speech Sect, NIH, Bethesda, MD 20892 USA. Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, NIH, Bethesda, MD USA. RP Ludlow, CL (reprint author), NINDS, Med Neurol Branch, Laryngeal & Speech Sect, NIH, 10 Ctr Dr,MSC 1416,Bldg 10,Room 5D38, Bethesda, MD 20892 USA. OI Ludlow, Christy/0000-0002-2015-6171 NR 22 TC 11 Z9 11 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0023-852X J9 LARYNGOSCOPE JI Laryngoscope PD NOV PY 2000 VL 110 IS 11 BP 1943 EP 1949 DI 10.1097/00005537-200011000-00033 PG 7 WC Medicine, Research & Experimental; Otorhinolaryngology SC Research & Experimental Medicine; Otorhinolaryngology GA 371EL UT WOS:000165165200032 PM 11081615 ER PT J AU Haga, SB Fu, S Karp, JE Ross, DD Williams, DM Hankins, WD Behm, F Ruscetti, FW Chang, M Smith, BD Becton, D Raimondi, SC Berg, PE AF Haga, SB Fu, S Karp, JE Ross, DD Williams, DM Hankins, WD Behm, F Ruscetti, FW Chang, M Smith, BD Becton, D Raimondi, SC Berg, PE TI BP1, a new homeobox gene, is frequently expressed in acute leukemias SO LEUKEMIA LA English DT Article DE homeobox; BP1; leukemia; clonogenicity; CD34 ID ACUTE MYELOID-LEUKEMIA; HUMAN HEMATOPOIETIC-CELLS; ACUTE LYMPHOBLASTIC-LEUKEMIA; TRANSCRIPTION FACTORS; C-MYB; CHROMOSOME-TRANSLOCATION; RESTRICTED EXPRESSION; HOX GENES; MLL; PROTEIN AB Aberrant expression of homeobox genes has been described in primary leukemia blasts. We recently cloned a new cDNA, BP1, which is a member of the homeobox gene family. BP1 expression was investigated in bone marrow samples from acute myeloid leukemia (AML), acute T cell lymphocytic leukemia (ALL) and pre-B cell ALL. Expression levels of two apparent isoforms of BP1, DLX7 and DLX4, were measured in the same samples. They are weakly if at all detectable in normal bone marrow, PHA-stimulated T cells or B cells. BP1 RNA was highly expressed in 63% of AML cases, including 81% of the pediatric and 47% of the adult cases, and in 32% of T-ALL cases, but was not found in any of the pre-B ALL cases. Coexpression of BP1, DLX7 and DLX4 occurred in a significant number of leukemias. Our data, including co-expression of BP1 with c-myb and GATA-1, markers of early progenitors, suggest that BP1 expression occurs in primitive cells in AML. Analysis of CD34(+) and CD34(-) normal bone marrow cells revealed BP1 is expressed in CD34- cells and virtually extinguished in CD34(+) cells. Ectopic expression of BP1 in the leukemia cell line K562 increased clonogenicity, consistent with a role for BP1 in leukemogenesis. The presence of BP1 RNA in leukemic blasts may therefore be a molecular marker for primitive cells and/or may indicate that BP1 is an important upstream factor in an oncogenic pathway. C1 George Washington Univ, Med Ctr, Dept Biochem & Mol Biol, Washington, DC 20037 USA. Univ Maryland, Sch Med, Div Human Genet, Baltimore, MD 21201 USA. Univ Maryland, Sch Med, Greenebaum Canc Ctr, Baltimore, MD 21201 USA. Dept Vet Affairs, Baltimore Vet Med Ctr, Baltimore, MD USA. Johns Hopkins Univ, Dept Med, Baltimore, MD USA. ProED Inc, Bethesda, MD USA. St Jude Childrens Res Hosp, Dept Pathol & Lab Med, Memphis, TN 38105 USA. NCI, Frederick Canc Res & Dev Ctr, Lab Leukocyte Biol, Frederick, MD USA. Univ Florida, Stat Off, Gainesville, FL USA. Johns Hopkins Univ, Johns Hopkins Oncol Ctr, Baltimore, MD USA. Univ Arkansas Med Sci, Dept Pediat, Little Rock, AR 72205 USA. RP Berg, PE (reprint author), George Washington Univ, Med Ctr, Dept Biochem & Mol Biol, Ross Bldg,Room 533,2300 Eye St NW, Washington, DC 20037 USA. FU NCI NIH HHS [CA-21765, 5P30CA06973]; NIDDK NIH HHS [R01 DK53533] NR 51 TC 38 Z9 47 U1 0 U2 2 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0887-6924 J9 LEUKEMIA JI Leukemia PD NOV PY 2000 VL 14 IS 11 BP 1867 EP 1875 DI 10.1038/sj.leu.2401912 PG 9 WC Oncology; Hematology SC Oncology; Hematology GA 370LU UT WOS:000165125200002 PM 11069021 ER PT J AU Ward, KM Balaban, RS AF Ward, KM Balaban, RS TI Determination of pH using water protons and chemical exchange dependent saturation transfer (CEST) SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE spin lattice relaxation rate; gadolinium; contrast agent; phosphate; MRI; MRS ID INTRACELLULAR PH; MAGNETIC-RESONANCE; ERYTHROCYTES; METABOLITES; H-1-NMR AB Solution pH was measured using water proton NMR via chemical exchange dependent saturation transfer (CEST) with selected chemical exchange sites. Several useful pH-sensitive proton chemical exchange agents were found: 5,6-dihydrouracil, 5-hydroxytryptophan, and a combination of 5-hydroxytryptophan and 2-imidazolidinethione. A ratiometric approach was developed that permitted pH determinations that were independent of water T-1 or exchange site concentration. Published 2000 Wiley-Liss, Inc.(dagger) C1 NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Balaban, RS (reprint author), NHLBI, Cardiac Energet Lab, NIH, Bldg 10,Room B1D-416,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Balaban, Robert/A-7459-2009 OI Balaban, Robert/0000-0003-4086-0948 NR 16 TC 195 Z9 200 U1 0 U2 32 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD NOV PY 2000 VL 44 IS 5 BP 799 EP 802 DI 10.1002/1522-2594(200011)44:5<799::AID-MRM18>3.0.CO;2-S PG 4 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 371DW UT WOS:000165163600018 PM 11064415 ER PT J AU Zuberi, AR Church, D AF Zuberi, AR Church, D TI Mouse chromosome 2 SO MAMMALIAN GENOME LA English DT Article C1 Pennington Biomed Res Ctr, Baton Rouge, LA 70808 USA. NIH, DHHS, Natl Lib Med, NCBI, Bethesda, MD 20892 USA. RP Zuberi, AR (reprint author), Pennington Biomed Res Ctr, 6400 Perkins Rd, Baton Rouge, LA 70808 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 944 EP 945 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300003 ER PT J AU Liddell, RA Meek, CE Mock, BA AF Liddell, RA Meek, CE Mock, BA TI Mouse chromosome 4 SO MAMMALIAN GENOME LA English DT Article C1 NCI, Genet Lab, DBS, NIH, Bethesda, MD 20892 USA. RP Mock, BA (reprint author), NCI, Genet Lab, DBS, NIH, Bldg 37,Room 2B-08,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 946 EP 947 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300005 ER PT J AU Kozak, CA Stephenson, D AF Kozak, CA Stephenson, D TI Mouse chromosome 5 SO MAMMALIAN GENOME LA English DT Article C1 NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. McLaughlin Res Inst Biomed Sci, Stowers Inst Med Res, Great Falls, MT 59405 USA. RP Kozak, CA (reprint author), NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 947 EP 947 PG 1 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300006 ER PT J AU Bryda, EC Noben-Trauth, K Bureau, JF AF Bryda, EC Noben-Trauth, K Bureau, JF TI Mouse chromosome 10 SO MAMMALIAN GENOME LA English DT Article C1 Marshall Univ, Sch Med, Dept Microbiol Mol Genet & Immunol, Huntington, WV 25704 USA. Natl Inst Deafness & Other Commun Disorders, NIH, Rockville, MD 20850 USA. Inst Pasteur, Unite Virus Lents, F-75724 Paris, France. RP Bryda, EC (reprint author), Marshall Univ, Sch Med, Dept Microbiol Mol Genet & Immunol, 1542 Spring Valley Dr, Huntington, WV 25704 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 951 EP 952 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300011 ER PT J AU Stephenson, DA Lueders, KK AF Stephenson, DA Lueders, KK TI Mouse chromosome 13 SO MAMMALIAN GENOME LA English DT Article C1 McLaughlin Res Inst, Great Falls, MT 59405 USA. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Stephenson, DA (reprint author), McLaughlin Res Inst, 1520 23rd St S, Great Falls, MT 59405 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 954 EP 954 PG 1 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300014 ER PT J AU Cabin, DE Reeves, RH AF Cabin, DE Reeves, RH TI Mouse chromosome 16 SO MAMMALIAN GENOME LA English DT Article C1 NHGRI, Genet Dis Res Branch, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Physiol, Baltimore, MD 21205 USA. RP Cabin, DE (reprint author), NHGRI, Genet Dis Res Branch, 49 Convent Dr,494B67,MSC 4472, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 955 EP 956 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300017 ER PT J AU Huppi, K Spear, B Siwarski, D Letts, V AF Huppi, K Spear, B Siwarski, D Letts, V TI Mouse chromosome 15 SO MAMMALIAN GENOME LA English DT Article C1 NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. Univ Kentucky, Dept Microbiol & Immunol, Lexington, KY 40536 USA. Jackson Lab, Bar Harbor, ME 04609 USA. RP Huppi, K (reprint author), NCI, Genet Lab, NIH, Bldg 37,Rm 2B-21, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD NOV PY 2000 VL 11 IS 11 BP 955 EP 955 PG 1 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 367HU UT WOS:000090056300016 ER PT J AU Cope, JU AF Cope, JU TI A viral etiology for Ewing's sarcoma SO MEDICAL HYPOTHESES LA English DT Article ID SOFT-TISSUE SARCOMAS; CHILDHOOD-CANCER; BONE CANCER; CHILDREN; EPIDEMIOLOGY; TUMOR; RISK AB Despite the finding of characteristic somatic mutations in the tumor tissue and efforts to identify risk factors, the etiology of Ewing's sarcoma (ES) is still unknown. ES is very different from other childhood bone cancers. it rarely occurs in the black population and has no animal model. Recently studies indicate that ES may have a neural, not mesenchymal, origin. It has a distinctive unimodal age-incidence peak at adolescence. Because its incidence curve pattern has a striking resemblance to that of DES-related clear cell adenocarcinoma of the vagina, an in utero exposure might be considered. Although in utero chemical and hormonal exposures have not been found to be associated with ES in epidemiologic studies, we suggest that its etiology could be an in utero viral infection. We hypothesize that the epidemiological characteristics of ES suggest an association with cytomegalovirus (CMV). (C) 2000 Harcourt Publishers Ltd. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Cope, JU (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS Room 7007, Bethesda, MD 20892 USA. NR 36 TC 10 Z9 10 U1 0 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0306-9877 J9 MED HYPOTHESES JI Med. Hypotheses PD NOV PY 2000 VL 55 IS 5 BP 369 EP 372 DI 10.1054/mehy.2000.1069 PG 4 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 378XK UT WOS:000165610500001 PM 11058415 ER PT J AU Keith, BD AF Keith, BD TI Ursodeoxycholic acid to inhibit the growth of hepatic metastases SO MEDICAL HYPOTHESES LA English DT Article ID TRANSFORMING GROWTH-FACTOR-BETA-1; INDUCED APOPTOSIS; CELL-DEATH; RECEPTOR SUPERFAMILY; RAT HEPATOCYTES; FACTOR BETA-1; BILE-SALTS; ANTI-FAS; EXPRESSION; LIVER AB Cancer can be resistant to apoptosis. If such cancers had apoptotic stimuli in their microenvironment, these stimuli might induce apoptosis in surrounding host cells. The majority of host cells at the peritumoral margins of liver metastases are undergoing apoptosis. Damage to the bile duct system may result in bile acid release, which may cause apoptosis in surrounding host cells. Metastatic cells may be releasing substances, such as transforming growth factor-beta 1 (TGF-beta 1), that cause apoptosis in surrounding host tissue. Ursodeoxycholic acid might inhibit the growth of hepatic metastases that are resistant to apoptotic stimuli such as bile acids and TGF-beta 1. Ursodeoxycholic acid decreases apoptosis caused by other bile acids and TGF-beta 1. Chemotherapy of hepatic metastases resistant to apoptosis might cause apoptosis more in peritumoral host cells than in cancer cells. Antiapoptotic therapy might be effective in cancer sensitive to apoptosis depending on its interactions with chemotherapy and tumor cells. (C) 2000 Harcourt Publishers Ltd. C1 NCI, Med Branch, Natl Naval Med Ctr, Bethesda, MD 20889 USA. RP Keith, BD (reprint author), NCI, Med Branch, Natl Naval Med Ctr, Bldg 8,Room 5101, Bethesda, MD 20889 USA. NR 35 TC 1 Z9 2 U1 0 U2 0 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0306-9877 J9 MED HYPOTHESES JI Med. Hypotheses PD NOV PY 2000 VL 55 IS 5 BP 379 EP 382 DI 10.1054/mehy.2000.1071 PG 4 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 378XK UT WOS:000165610500003 PM 11058416 ER PT J AU Roy, PK Venzon, DJ Shojamanesh, H Abou-Saif, A Peghini, P Doppman, JL Gibril, F Jensen, RT AF Roy, PK Venzon, DJ Shojamanesh, H Abou-Saif, A Peghini, P Doppman, JL Gibril, F Jensen, RT TI Zollinger-Ellison syndrome - Clinical presentation in 261 patients SO MEDICINE LA English DT Review ID MULTIPLE ENDOCRINE NEOPLASIA; SOMATOSTATIN-RECEPTOR SCINTIGRAPHY; ISLET-CELL TUMORS; GASTRIC-ACID HYPERSECRETION; SECRETIN INJECTION TEST; LONG-TERM FOLLOW; SERUM GASTRIN; CHRONIC-PANCREATITIS; HELICOBACTER-PYLORI; BARRETTS-ESOPHAGUS C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. NCI, Biostat & Data Management Sect, Bethesda, MD 20892 USA. NIH, Warren Grant Magnuson Clin Ctr, Dept Radiol, Bethesda, MD 20892 USA. RP Jensen, RT (reprint author), NIDDKD, Digest Dis Branch, NIH, Bldg 10,Rm 9C-103,10 Ctr Dr,MSC 1804, Bethesda, MD 20892 USA. RI Venzon, David/B-3078-2008 NR 265 TC 161 Z9 167 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0025-7974 EI 1536-5964 J9 MEDICINE JI Medicine (Baltimore) PD NOV PY 2000 VL 79 IS 6 BP 379 EP 411 DI 10.1097/00005792-200011000-00004 PG 33 WC Medicine, General & Internal SC General & Internal Medicine GA 380LD UT WOS:000165702600004 PM 11144036 ER PT J AU Casanova, JL Holland, SM AF Casanova, JL Holland, SM TI Current focus: genetic susceptibility to bacterial infection: Mendelian predisposition to mycobacterial infection in man - Introduction SO MICROBES AND INFECTION LA English DT Editorial Material C1 Necker Med Sch, Lab Human Genet Infect Dis, F-75015 Paris, France. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Casanova, JL (reprint author), Necker Med Sch, Lab Human Genet Infect Dis, 156 Rue Vaugirard, F-75015 Paris, France. NR 0 TC 0 Z9 0 U1 0 U2 0 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 1286-4579 J9 MICROBES INFECT JI Microbes Infect. PD NOV PY 2000 VL 2 IS 13 BP 1547 EP 1547 DI 10.1016/S1286-4579(00)01309-5 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 380WY UT WOS:000165727600001 PM 11113372 ER PT J AU Holland, SM AF Holland, SM TI Treatment of infections in the patient with Mendelian susceptibility to mycobacterial infection SO MICROBES AND INFECTION LA English DT Article DE IFN-gamma; IFN-alpha; IL-12; GM-CSF; TNF ID TUMOR-NECROSIS-FACTOR; COLONY-STIMULATING FACTOR; MULTIDRUG-RESISTANT TUBERCULOSIS; RECOMBINANT INTERFERON-GAMMA; AVIUM COMPLEX INFECTION; CELL-MEDIATED-IMMUNITY; ACTIVATES HUMAN MACROPHAGES; ERYTHEMA-NODOSUM LEPROSUM; BACILLE CALMETTE-GUERIN; JAK-STAT PATHWAY AB Cytokines are increasingly used for the therapy of infections in patient populations with special defects in immunity (chemotherapy, bone marrow transplantation, chronic granulomatous disease). The recognition of multiple defects in the systems of the interferon-gamma (IFN-gamma) receptor, interleukin-12 (IL-12) receptor and IL-12 p40 emphasizes the critical roles that cytokines play in preventing and clearing infection. The cases of patients with partially responsive IFN-gamma receptors (autosomal dominant and partial defects) are ideal candidates for successful cytokine prophylaxis and therapy. Better understanding of the critical elements of the cytokine pathways may show us ways to circumvent these defects with complementary cytokine cascades. (C) 2000 NIH. Published by Editions scientifiques et medicales Elsevier SAS. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Holland, SM (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,11N103,10 Ctr Dr,MSC 1886, Bethesda, MD 20892 USA. NR 98 TC 19 Z9 21 U1 0 U2 1 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 1286-4579 J9 MICROBES INFECT JI Microbes Infect. PD NOV PY 2000 VL 2 IS 13 BP 1579 EP 1590 DI 10.1016/S1286-4579(00)01314-9 PG 12 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 380WY UT WOS:000165727600006 PM 11113377 ER PT J AU Kamhawi, S AF Kamhawi, S TI The biological and immunomodulatory properties of sand fly saliva and its role in the establishment of Leishmania infections SO MICROBES AND INFECTION LA English DT Review DE sand fly saliva; Leishmania; immunomodulation; infectivity; presensitization ID CYCLASE-ACTIVATING PEPTIDE; NITRIC-OXIDE PRODUCTION; CHEMOKINE GENE-EXPRESSION; I RECEPTOR AGONIST; LUTZOMYIA-LONGIPALPIS; VECTOR SALIVA; PHLEBOTOMUS-PAPATASI; PARASITE TRANSMISSION; VASODILATORY PEPTIDE; MOUSE MACROPHAGES AB Sand fly saliva contains a rich array of pharmacologically active compounds whose primary function is to prevent the hemostatic mechanisms of the host. Several studies have ascribed immunosuppressive properties to sand fly saliva as well as an exacerbative effect on Leishmania infectivity for their mammalian hosts. This review provides a comprehensive account of sand fly salivary components, the immunomodulatory properties exhibited by some of its molecules, and describes che findings concerning the influence of saliva on Leishmania infections. The potential use of saliva as part of an anti-Leishmania vaccine for the mammalian host is also addressed. (C) 2000 Editions scientifiques et medicales Elsevier SAS. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Kamhawi, S (reprint author), NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. NR 73 TC 80 Z9 82 U1 0 U2 2 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 1286-4579 J9 MICROBES INFECT JI Microbes Infect. PD NOV PY 2000 VL 2 IS 14 BP 1765 EP 1773 DI 10.1016/S1286-4579(00)01331-9 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 391DJ UT WOS:000166338600010 PM 11137049 ER PT J AU Leak, LV Saunders, M Day, AA Jones, M AF Leak, LV Saunders, M Day, AA Jones, M TI Stimulation of plasminogen activator and inhibitor in the lymphatic endothelium SO MICROVASCULAR RESEARCH LA English DT Article DE lymphatic endothelium; tissue plasminogen activator, tPA; plasminogen inhibitor; PAI-1; tPA mRNA; PIA-1 mRNA ID PROTEIN-KINASE-C; CLINICAL ASPECTS; TISSUE-TYPE; UROKINASE; CELLS; GENE; IDENTIFICATION; FIBRINOLYSIS; EXPRESSION; RELEASE AB Chromogenic assays, immunoblotting, and Northern blot hybridization methods were employed to assess the effects of various agonists on the production of tissue plasminogen activator (t similar to PA) and plasminogen activator inhibitor type 1 (PAI-1) by the lymphatic endothelium (LEC). Fibrin autography showed that plasminogen-dependent fibrinolytic activity occurred at M-r of 110 kDa, which represents a complex of tPA with PAI-1, and 65- and 55-kDa bands corresponding to tPA and uPA, respectively. The fractionation of lymph collected from ovine lymphatic vessels also produced a prominent lytic band of similar to 110 kDa, suggesting the formation of PA/PAI complexes in lymph. The stimulationn of various agonists produced large-scale increases in tPA mRNA, as shown by Northern blot hybridization analyses. The effects of ECGF, histamine, and LPS on the presence of tPA and on enhancing the levels of mRNA reached maximum activity at 4 h and declined to levels below that of controls by 8 h. However, phorbol-treated cells exhibited reduced levels of tPA mRNA at 4 h, but was significantly increased by 8 h. A large-scale increase in PAI-1 mRNA steady-state levels was also stimulated by the agonists used in these studies. Both the 3.4- and 2.4-kb species of PAI-1 mRNA were increased. These observations demonstrated that tPA and PAI-1 are produced and secreted by LEC monolayer cultures and are also present in lymph. (C) 2000 Academic Press. C1 Howard Univ, Coll Med, E E Just Lab Cellular Biol, Washington, DC 20059 USA. Boston Univ, Dept Orthoped Surg, Boston, MA 02118 USA. Howard Univ, Coll Med, Ctr Canc, Washington, DC 20059 USA. Howard Univ, Dept Microbiol, Washington, DC 20059 USA. NHLBI, Lab Anim Med & Surg, Bethesda, MD 20892 USA. RP Leak, LV (reprint author), Howard Univ, Coll Med, E E Just Lab Cellular Biol, Washington, DC 20059 USA. FU NIGMS NIH HHS [S06GM08016] NR 38 TC 9 Z9 9 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0026-2862 J9 MICROVASC RES JI Microvasc. Res. PD NOV PY 2000 VL 60 IS 3 BP 201 EP 211 DI 10.1006/mvre.2000.2262 PG 11 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 374KA UT WOS:000165343300002 PM 11078636 ER PT J AU Lin, SK Wang, WG Wilson, GM Yang, XL Brewer, G Holbrook, NJ Gorospe, M AF Lin, SK Wang, WG Wilson, GM Yang, XL Brewer, G Holbrook, NJ Gorospe, M TI Down-regulation of cyclin D1 expression by prostaglandin A(2) is mediated by enhanced cyclin D1 mRNA turnover SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID MESSENGER-RNA DEGRADATION; AU-RICH ELEMENT; 3' UNTRANSLATED REGION; CELL-CYCLE; 3'-UNTRANSLATED REGION; CANCER-CELLS; ANTINEOPLASTIC PROSTAGLANDINS; RETINOBLASTOMA PROTEIN; MAMMALIAN-CELLS; BINDING PROTEIN AB Prostaglandin A(2) (PGA(2)), an experimental chemotherapeutic agent, causes growth arrest associated with decreased cyclin D1 expression in several cancer cell lines. Here, using human non-small-cell lung carcinoma H1299 cells, we investigated the mechanisms whereby PGA(2) down-regulates cyclin D1 expression. Transcription rates of the cyclin D1 gene, studied using a cyclin D1 promoter-luciferase construct and nuclear run-on assays, were not affected by PGA(2) treatment. Instead, the cyclin D1 mRNA was rendered unstable after exposure to PGA(2). Since the stability of labile mRNA is modulated through binding of proteins to specific mRNA sequences, we sought to identify protein(s) recognizing the cyclin D1 mRNA. In electrophoretic mobility-shift assays using radiolabeled RNA probes derived from different regions of cyclin D1 mRNA,we observed that (i) lysates prepared from PGA(2)-treated cells exhibited enhanced protein-cyclin D1 RNA complex formation; (ii) the kinetics of complex formation correlated closely,vith that of cyclin D1 mRNA loss; and (iii) binding occurred within a 390-base cyclin D1 3' untranslated region (UTR) (K12). This binding activity could be cross-linked, revealing proteins ranging from 30 to 37 kDa. The RNA-binding protein AUF1, previously associated with the degradation of target mRNAs, bound cyclin D1 mRNA, because anti-AUF1 antibodies were capable of supershifting or immunoprecipitating cyclin D1 mRNA protein complexes. Finally, insertion of K12 in the 3'UTR of reporter genes markedly reduced the expression and half-life of the resulting chimeric mRNAs in transfected, PGA(2)-treated cells. Our data demonstrate that PGA(2) down-regulates cyclin D1 expression by decreasing cyclin D1 mRNA stability and implicates a 390-base element in the 3'UTR in this regulation. C1 NIA, Biol Chem Lab, GRC, NIH, Baltimore, MD 21224 USA. Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Mol Genet & Microbiol, Piscataway, NJ 08854 USA. RP Gorospe, M (reprint author), NIA, Biol Chem Lab, GRC, NIH, Box 12,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Lin, Shan-Kung/B-1062-2010 FU NCI NIH HHS [R01 CA052443, R01CA52443] NR 64 TC 109 Z9 112 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 NOV PY 2000 VL 20 IS 21 BP 7903 EP 7913 DI 10.1128/MCB.20.21.7903-7913.2000 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 362TK UT WOS:000089794200007 PM 11027261 ER PT J AU Avedisov, SN Krasnoselskaya, I Mortin, M Thomas, BJ AF Avedisov, SN Krasnoselskaya, I Mortin, M Thomas, BJ TI Roughex mediates G(1) arrest through a physical association with cyclin A SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID DEPENDENT-KINASE INHIBITOR; CELL-CYCLE; S-PHASE; DROSOPHILA CYCLIN; PATTERN-FORMATION; MITOTIC CYCLINS; DOWN-REGULATION; PROTEOLYSIS; EXPRESSION; PROTEIN AB Differentiation in the developing Drosophila eye requires synchronization of cells in the G(1) phase of the cell cycle. The rougher gene product plays a key role in this synchronization by negatively regulating cyclin A protein levels in G(1). We show here that coexpressed Rougher and cyclin A physically interact in viva, Rougher is a nuclear protein, while cyclin A was previously shown to be exclusively cytoplasmic during interphase in the embryo. In contrast, we demonstrate that in interphase cells in the eye imaginal disk cyclin A Is present in both the nucleus and the cytoplasm. In the presence of ectopic Rougher, cyclin A becomes strictly nuclear and is later degraded. Nuclear targeting of both Rougher and cyclin A under these conditions is dependent on a C-terminal nuclear localization signal in Rougher. Disruption of this signal results in cytoplasmic localization of both Rougher and cyclin it confirming a physical interaction between these molecules. Cyclin A interacts with both Cdc2 and Cdc2c, the Drosophila Cdk2 homolog, and Rougher inhibits the histone H1 kinase activities of both cyclin A-Cdc2 and cyclin A-Cdc2c complexes in whole-cell extracts. Two-hybrid experiments suggested that the inhibition of kinase activity by Rougher results from competition with the cyclin-dependent kinase subunit for binding to cyclin A. These findings suggest that Rougher can influence the intracellular distribution of cyclin A and define Rougher as a distinct and specialized cell cycle inhibitor for cyclin A-dependent kinase activity. C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Thomas, BJ (reprint author), NCI, Biochem Lab, NIH, Bldg 37,Room 4C10,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. RI Mortin, Mark/B-4251-2008 NR 43 TC 24 Z9 24 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 NOV PY 2000 VL 20 IS 21 BP 8220 EP 8229 DI 10.1128/MCB.20.21.8220-8229.2000 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 362TK UT WOS:000089794200037 PM 11027291 ER PT J AU Li, M Damania, B Alvarez, X Ogryzko, V Ozato, K Jung, JU AF Li, M Damania, B Alvarez, X Ogryzko, V Ozato, K Jung, JU TI Inhibition of p300 histone acetyltransferase by viral interferon regulatory factor SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID SARCOMA-ASSOCIATED HERPESVIRUS; KAPOSIS-SARCOMA; TRANSCRIPTIONAL ACTIVATION; ONCOPROTEIN E1A; ENCODES; COACTIVATOR; CBP; CBP/P300; PROTEIN; HOMOLOG AB Kaposi's sarcoma-associated herpesvirus (KSHV) has been consistently identified in Kaposi's sarcomas, body cavity-based lymphomas, and some forms of Castleman's disease. The K9 open reading frame of KSHV encodes a viral interferon regulatory factor (vIRF) which functions as a repressor far cellular interferon-mediated signal transduction and as an oncogene to induce cell gran th transformation. We demonstrate that KSHV vIRF directly interacts with cellular transcriptional coactivator p300 and displaces p300/CBP-associated factor from p300 complexes. This interaction inhibits the histone acetyltransferase activity of p300, resulting in drastic reduction of nucleosomal histone acetylation and alteration of chromatin structure. As a consequence, vIRF expression markedly alters cellular cytokine expression, which is regulated by acetylation of nucleosomal histones. These results demonstrate that KSHV vIRF interacts with and inhibits the p300 transcriptional coactivator to circumvent the host antiviral immune response and to induce a global alteration of cellular gene expression. These studies also illustrate how a cellular gene captured by a herpesvirus has evolved several functions that suit the needs of the virus. C1 Harvard Univ, Sch Med, New England Reg Primate Res Ctr, Dept Microbiol & Mol Genet, Southborough, MA 01772 USA. Harvard Univ, Sch Med, New England Reg Primate Res Ctr, Dept Pathol, Southborough, MA 01772 USA. NICHD, Lab Mol Growth Regulat, Bethesda, MD 20892 USA. RP Jung, JU (reprint author), Harvard Univ, Sch Med, New England Reg Primate Res Ctr, Dept Microbiol & Mol Genet, 1 Pine Hill Dr, Southborough, MA 01772 USA. RI Ogryzko, Vasily/M-6665-2015 OI Ogryzko, Vasily/0000-0002-8548-1389 FU NCI NIH HHS [CA31363, CA82057, CA86841, R01 CA031363, R01 CA082057]; NCRR NIH HHS [K26 RR000168, P51 RR000168] NR 52 TC 96 Z9 101 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 NOV PY 2000 VL 20 IS 21 BP 8254 EP 8263 DI 10.1128/MCB.20.21.8254-8263.2000 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 362TK UT WOS:000089794200040 PM 11027294 ER PT J AU Nakazato, M Chung, HK Ulianich, L Grassadonia, A Suzuki, K Kohn, LD AF Nakazato, M Chung, HK Ulianich, L Grassadonia, A Suzuki, K Kohn, LD TI Thyroglobulin repression of thyroid transcription factor 1 (TTF-1) gene expression is mediated by decreased DNA binding of nuclear factor I proteins which control constitutive TTF-1 expression SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID TISSUE-SPECIFIC EXPRESSION; THYROTROPIN RECEPTOR GENE; NA+/I SYMPORTER GENE; FOLLICULAR THYROGLOBULIN; POTENTIAL ROLE; PROMOTER REGION; REPLICATION; SITE; ENHANCER; CELLS AB Follicular thyroglobulin (TG) selectively suppresses the expression of thyroid-restricted transcription factors, thereby altering the expression of thyroid-specific proteins. In this study, we investigated the molecular mechanism by which TG suppresses the prototypic thyroid-restricted transcription factor, thyroid transcription factor 1 (TTF-1), in rat FRTL-5 thyrocytes. We show that the region between bp -264 and -153 on the TTF-1 promoter contains two nuclear factor I (NFI) elements whose function is involved in TG-mediated suppression. Thus, NFI binding to these elements is critical for constitutive expression of TTF-1; TG decreases NFI binding to the NFI elements in association with TG repression. NFI is a family of transcription factors that is ubiquitously expressed and contributes to constitutive and cell-specific gene expression. In contrast to the contribution of NFI proteins to constitutive gene expression in other systems, we demonstrate that follicular TG transcriptionally represses all NFI RNAs (NFI-A, -B, -C, and -X) in association with decreased NFI binding and that the RNA levels decrease as early as 4 h after TG treatment. Although TG treatment for 48 h results in a decrease in NFI protein-DNA complexes measured in DNA mobility shift assays, NFI proteins are still detectable by Western analysis. We show, however, that the binding of all NFI proteins is redox regulated. Thus, diamide treatment of nuclear extracts strongly reduces the binding of NFI proteins, and the addition of higher concentrations of dithiothreitol to nuclear extracts from TG-treated cells restores NFI-DNA binding to levels in extracts from untreated cells. We conclude that NFI binding to two NFI elements, at bp -265 to -153, positively regulates TTF-1 expression and controls constitutive TTF-1 levels. TG mediates the repression of TTF-1 gene expression by decreasing NFI RNA and protein levels, as well as by altering the binding activity of NFI, which is redox controlled. C1 NIDDK, Cell Regulat Sect, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Kohn, LD (reprint author), NIDDK, Cell Regulat Sect, Metab Dis Branch, NIH, Bldg 10,Room 9C101B, Bethesda, MD 20892 USA. NR 59 TC 14 Z9 15 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 NOV PY 2000 VL 20 IS 22 BP 8499 EP 8512 DI 10.1128/MCB.20.22.8499-8512.2000 PG 14 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 368AH UT WOS:000090094800019 PM 11046146 ER PT J AU Nicot, C Harrod, R AF Nicot, C Harrod, R TI Distinct p300-responsive mechanisms promote caspase dependent apoptosis by human T-cell lymphotropic virus type 1 Tax protein SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID CREB-BINDING-PROTEIN; NF-KAPPA-B; TRANSCRIPTIONAL INTEGRATOR P300; LEUKEMIA-VIRUS; GLUCOCORTICOID RECEPTOR; TRANSFORMING PROTEIN; NUCLEAR INTEGRATION; TRANSGENIC MICE; COACTIVATOR CBP; C-JUN AB The dysregulation of cellular apoptosis pathways has emerged as a critical early event associated with the development of many types of human cancers. Numerous viral and cellular oncogenes, aside from their inherent transforming properties, are known to induce programmed cell death, consistent with the hypothesis that genetic defects are required to support tumor survival. Here, we report that nuclear expression of the CREB-binding protein (CBP)/p300-binding domain of the human T-cell lymphotropic virus type 1 (HTLV-1) transactivator, Tax, triggers an apoptotic death-inducing signal during short-term clonal analyses, as well as in transient cell death assays. Coexpression of the antiapoptotic factor Bcl-2 increased serum stimulation; incubation with the chemical caspase inhibitor z-Val-Ala-DL-Asp fluoromethylketone antagonized Tax-induced cell death. The CBP/p300-binding defective Tax mutants K88A and V89A exhibited markedly reduced cytotoxic effects compared to the wild-type Tax protein. Importantly, nuclear expression of the minimal CBP/p300-binding peptide of Tax induced apoptosis in the absence of Tax-dependent transcriptional activities, while its K88A counterpart did not cause cell death. Further, Tax-mediated apoptosis was effectively prevented by ectopic expression of the p300 coactivator. We also report that activation of the NF-kappaB transcription pathway by Tax, under growth arrest conditions, results in apoptosis that occurs independent of direct Tax coactivator effects. Our results allude to a novel pivotal role for the transcriptional coactivator p300 in determining cell fate and raise the possibility that dysregulated coactivator usage may pose an early barrier to transformation that must be selectively overcome as a prerequisite for the initiation of neoplasia. C1 NCI, Basic Res Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Nicot, C (reprint author), NCI, Basic Res Lab, Div Basic Sci, NIH, Bldg 41,Room D804,9000 Rockville Pike, Bethesda, MD 20892 USA. FU NCI NIH HHS [R01CA48709, R01 CA048709, R01CA/GM75688, R01 CA075688]; NIGMS NIH HHS [F32 GM075688] NR 60 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 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD NOV PY 2000 VL 20 IS 22 BP 8580 EP 8589 DI 10.1128/MCB.20.22.8580-8589.2000 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 368AH UT WOS:000090094800026 PM 11046153 ER PT J AU Li, JJ Cao, Y Young, MR Colburn, NH AF Li, JJ Cao, Y Young, MR Colburn, NH TI Induced expression of dominant-negative c-jun downregulates NF kappa B and AP-1 target genes and suppresses tumor phenotype in human keratinocytes SO MOLECULAR CARCINOGENESIS LA English DT Article DE nuclear factor kappa B; activator protein 1; neoplastic progression; human keratinocytes; inducible transgene ID MOUSE SKIN CARCINOGENESIS; INDUCED TRANSFORMATION; RETINOIC ACID; TRANSCRIPTION FACTORS; HUMAN STROMELYSIN; MAMMALIAN-CELLS; TRANSGENIC MICE; MESSENGER-RNA; ACTIVATION; MUTANT AB Neoplastically transformed mouse and human keratinocytes elevate transactivation of both activator protein 1 (AP-1) and nuclear factor kappaB (NF kappaB) transcription factors. The present study addresses the question of whether elevated NF kappaB in addition to elevated AP-1-dependent gene expression is necessary for maintaining the tumor cell phenotype. When a tetracycline-regulatable dominant-negative c-jun (TAM67, having a truncated transactivation domain) was expressed in tumorigenic human keratinocytes, AP-1- and NF kappaB- but not p53-dependent reporter activity was inhibited by 40-60%. Tumor phenotype, as measured by anchorage-independent growth, was inhibited by 90%. Neither AP-1/NF kappaB activation nor expression of tumor phenotype was inhibited in TAM67-harboring keratinocytes under noninducing conditions. Electrophoretic mobility shift analysis showed that induction of TAM67 expression slightly increased AP-1- but reduced NF kappaB DNA-binding activity. Immunoprecipitation showed that TAM67 interacted in keratinocyte nuclei with NF kappaB p65, suggesting that inhibition of NF kappaB by TAM67 is mediated by direct protein-protein interactions, possibly producing decreased binding to DNA or inactivating p65. To analyze the putative effector genes that may be targeted by TAM67, expression of genes responsive to AP-1 or NF kappaB was measured by reverse transcriptase-polymerase chain reaction in TAM67 transfectants with or without TAM67 induction. Induction of TAM67 inhibited or reduced the expression of collagenase I, stromelysin I (AP-1 responsive), and interleukins 1 and 6 (NF kappaB responsive). These results indicate that genes controlled by NF kappaB and by AP-1 may be transformation-relevant targets of TAM67 and that TAM67 may inhibit NF kappaB activation through direct interaction with NF kappaB p65. Moreover, the findings provide proof for the principle of using inducible TAM67 as a gene therapy to suppress tumor phenotype in human carcinoma cells. Published 2000 Wiley-Liss. Inc. C1 NCI, Frederick Canc Res & Dev Ctr, Lab Biochem Physiol, Gene Regulat Sect,NIH, Frederick, MD 21702 USA. RP Colburn, NH (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Biochem Physiol, Gene Regulat Sect,NIH, Box B,Bldg 560, Frederick, MD 21702 USA. NR 53 TC 67 Z9 68 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 NOV PY 2000 VL 29 IS 3 BP 159 EP 169 DI 10.1002/1098-2744(200011)29:3<159::AID-MC5>3.0.CO;2-W PG 11 WC Biochemistry & Molecular Biology; Oncology SC Biochemistry & Molecular Biology; Oncology GA 379PE UT WOS:000165651000005 PM 11108661 ER PT J AU Romanienko, PJ Camerini-Otero, RD AF Romanienko, PJ Camerini-Otero, RD TI The mouse Spo11 gene is required for meiotic chromosome synapsis SO MOLECULAR CELL LA English DT Article ID SYNAPTONEMAL COMPLEXES; YEAST MEIOSIS; RECOMBINATION; DMC1; MICE; SPERMATOGENESIS; HOMOLOGS; FAMILY; RAD51; FORM AB The Spell protein initiates meiotic recombination by generating DNA double-strand breaks (DSBs) and is required for meiotic synapsis in S. cerevisiae. Surprisingly, Spoil homologs are dispensable for synapsis in C. elegans and Drosophila yet required for meiotic recombination. Disruption of mouse Spell results in infertility. Spermatocytes arrest prior to pachytene with little or no synapsis and undergo apoptosis. We did not detect Rad51/Dmc1 foci in meiotic chromosome spreads, indicating DSBs are not formed. Cisplatin-induced DSBs restored Rad51/Dmc1 foci and promoted synapsis. Spell localizes to discrete foci during leptotene and to homologously synapsed chromosomes. Other mouse mutants that arrest during meiotic prophase (Atm (-/-), Dmc1 (-/-), mei1, and Morc (-/-)) showed altered Spell protein localization and expression. We speculate that there is an additional role for Spell, after it generates DSBs, in synapsis. C1 NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. RP Camerini-Otero, RD (reprint author), NIDDKD, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. NR 51 TC 401 Z9 410 U1 4 U2 17 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell. PD NOV PY 2000 VL 6 IS 5 BP 975 EP 987 DI 10.1016/S1097-2765(00)00097-6 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 376TD UT WOS:000165472100001 PM 11106738 ER PT J AU Sarno, RJ David, VA Franklin, WL O'Brien, SJ Johnson, WE AF Sarno, RJ David, VA Franklin, WL O'Brien, SJ Johnson, WE TI Development of microsatellite markers in the guanaco, Lama guanicoe: utility for South American camelids SO MOLECULAR ECOLOGY LA English DT Article DE artiodactyl; guanaco; microsatellite; South American camelid ID DNA C1 NCI, Lab Genome Divers, FCRDC, Frederick, MD 21702 USA. Iowa State Univ, Dept Anim Ecol, Ames, IA 50011 USA. RP Sarno, RJ (reprint author), NCI, Lab Genome Divers, FCRDC, Frederick, MD 21702 USA. RI Johnson, Warren/D-4149-2016 OI Johnson, Warren/0000-0002-5954-186X NR 5 TC 26 Z9 26 U1 0 U2 3 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0962-1083 J9 MOL ECOL JI Mol. Ecol. PD NOV PY 2000 VL 9 IS 11 BP 1922 EP 1924 DI 10.1046/j.1365-294x.2000.01077-3.x PG 3 WC Biochemistry & Molecular Biology; Ecology; Evolutionary Biology SC Biochemistry & Molecular Biology; Environmental Sciences & Ecology; Evolutionary Biology GA 375MX UT WOS:000165404800026 PM 11091331 ER PT J AU Kawamoto, T Kakizaki, S Yoshinari, K Negishi, M AF Kawamoto, T Kakizaki, S Yoshinari, K Negishi, M TI Estrogen activation of the nuclear orphan receptor CAR (Constitutive active receptor) in induction of the mouse Cyp2b10 gene SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID RESPONSIVE ENHANCER MODULE; PHENOBARBITAL RESPONSIVENESS; LIVER AB The nuclear orphan receptor CAR (constitutively active receptor or constitutive androstane receptor) can be activated in response to xenochemical exposure, such as activation by phenobarbital of a response element called NR1 found in the CYP2B gene. Here various steroids were screened for potential endogenous chemicals that may activate CAR, using the NR1 enhancer and Cyp2b10 induction in transfected HepG2 cell and/or in mouse primary hepatocytes as the experimental criteria. 17 beta -Estradiol and estrone activated NR1, whereas estriol, estetrol, estradiol sulfate, and the synthetic estrogen diethylstilbestrol did not. On the other hand, progesterone and androgens repressed NR1 activity in HepG2 cells, and the repressed NR1 activity was fully restored by estradiol. Moreover, estrogen treatment elicited nuclear accumulation of CAR in the mouse livers, as well as primary hepatocytes, and induced the endogenous Cyp2b10 gene. Ovariectomy did not affect either the basal or induced level of CAR in the nucleus of the female livers, while castration slightly increased the basal and greatly increased the induced levels in the liver nucleus of male mice. Thus, endogenous estrogen appears not to regulate CAR in female mice, whereas endogenous androgen may be the repressive factor in male mice. Estrogen at pharmacological levels is an effective activator of CAR in both female and male mice, suggesting a biological and/or toxicological role of this receptor in estrogen metabolism. In addition to mouse CAR, estrogens activated rat CAR, whereas human CAR did not respond well to the estrogens under the experimental conditions. 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. RI Kawamoto, Takeshi/D-7938-2015 OI Kawamoto, Takeshi/0000-0003-3337-1775 NR 19 TC 111 Z9 118 U1 0 U2 5 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 NOV PY 2000 VL 14 IS 11 BP 1897 EP 1905 DI 10.1210/me.14.11.1897 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 368UR UT WOS:000090136300015 PM 11075820 ER PT J AU Akyurek, LM Yang, ZY Aoki, K San, H Nabel, GJ Parmacek, MS Nabel, EG AF Akyurek, LM Yang, ZY Aoki, K San, H Nabel, GJ Parmacek, MS Nabel, EG TI SM22 alpha promoter targets gene expression to vascular smooth muscle cells in vitro and in vivo SO MOLECULAR MEDICINE LA English DT Article ID HEAVY-CHAIN GENE; TRANSGENIC MICE; ARTERIAL; ATHEROSCLEROSIS; PROTEINS; REGION AB Background: Gene transfer into vascular smooth muscle cells (vsmcs) holds promise for studying the pathogenesis of arterial disorders. However, a potential limitation of vectors with heterologous promoters is organ toxicity resulting from unrestricted transgene expression. Vascular smooth muscle cell-specific gene expression could increase the safety of vectors for vascular diseases. Materials and Methods: To develop vectors that target gene expression to vsmcs, we constructed vectors encoding, human placental alkaline phosphatase (hpAP) and chloramphenicol transferase (CAT) driven by a 441-bp region of the murine SM22 alpha promoter (AdSM22 alpha -hpAP). Results: Transfection of AdSM22 alpha -hpAP into vascular and nonvascular cells resulted in the expression of alkaline phosphatase (AP) in primary arterial and venous smcs, but not in primary endothelial cells or National Institutes of Health (NIH) 3T3 cells. Expression of AP was observed on 32.5 +/- 1.4% of primary pig vsmcs-infected AdSM22 alpha -hpAP at a multiplicity of infection (MOI) of 500; whereas, infection with AdCMV-hpAP resulted in 100 +/- 0.0% expression at a MOI of 250. In vitro, expression from the heterologous cytomegalovirus (CMV) promoter was approximately 10(3)-fold higher in vsmcs, compared with the SM22 alpha promoter. Following introduction of AdSM22 alpha -hpAP vectors into balloon-injured pig arteries, AP recombinant protein was detected in neointimal (2.23 +/- 1.14%) and medial (0.56 +/- 0.21%) smcs, but not in endothelial or adventitial cells. In contrast, AdCMV-hpAP vectors led to AP expression in intimal endothelial and smcs cells (39.14 +/- 10.09%) and medial smcs (2.84 +/- 1.05%). AP expression was not observed in endothelial or vsmcs following transfection with the control vector, adenoviral vector lacking E1 (Ad Delta E1). Conclusions: The SM22 alpha promoter programs recombinant gene expression exclusively to vascular smcs in vitro and in vivo. Although expression levels are lower than with heterologous promoters, these vectors may provide a safe and effective tool for gene therapy of vascular diseases. C1 NHLBI, Vasc Biol Branch, NIH, Bethesda, MD 20892 USA. NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA. Univ Penn, Sch Med, Dept Med, Philadelphia, PA 19104 USA. RP Nabel, EG (reprint author), NHLBI, Vasc Biol Branch, NIH, Bldg 10,Room 8C103, Bethesda, MD 20892 USA. FU PHS HHS [R01 56915] NR 24 TC 17 Z9 18 U1 1 U2 2 PU JOHNS HOPKINS UNIV PRESS PI BALTIMORE PA JOURNALS PUBLISHING DIVISION, 2715 NORTH CHARLES ST, BALTIMORE, MD 21218-4319 USA SN 1076-1551 J9 MOL MED JI Mol. Med. PD NOV PY 2000 VL 6 IS 11 BP 983 EP 991 PG 9 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 379CX UT WOS:000165624300006 PM 11147575 ER PT J AU Slayden, RA Lee, RE Barry, CE AF Slayden, RA Lee, RE Barry, CE TI Isoniazid affects multiple components of the type II fatty acid synthase system of Mycobacterium tuberculosis SO MOLECULAR MICROBIOLOGY LA English DT Article ID CATALASE-PEROXIDASE GENE; MYCOLIC ACIDS; OXIDATIVE-STRESS; ESCHERICHIA-COLI; RESISTANCE; TARGET; INHA; TRICLOSAN; THIOLACTOMYCIN; BIOSYNTHESIS AB Genetic and biochemical evidence has implicated two different target enzymes for isoniazid (INH) within the unique type II fatty acid synthase (FAS) system involved in the production of mycolic acids. These two components are an enoyl acyl carrier protein (ACP) reductase, InhA, and a beta -ketoacyl-ACP synthase, KasA. We compared the consequences of INH treatment of Mycobacterium tuberculosis (MTB) with two inhibitors having well-defined targets: triclosan (TRC), which inhibits InhA; and thiolactomycin (TLM), which inhibits KasA. INH and TLM, but not TRC, upregulate the expression of an operon containing five FAS II components, including kasA and acpM. Although all three compounds inhibit mycolic acid synthesis, treatment with INH and TLM, but not with TRC, results in the accumulation of ACP-bound lipid precursors to mycolic acids that were 26 carbons long and fully saturated. TLM-resistant mutants of MTB were more cross-resistant to INH than TRC-resistant mutants. Overexpression of KasA conferred more resistance to TLM and INH than to TRC. Overexpression of InhA conferred more resistance to TRC than to INH and TLM. Co-overexpression of both InhA and KasA resulted in strongly enhanced levels of INH resistance, in addition to cross-resistance to both TLM and TRC. These results suggest that these components of the FAS II complex are not independently regulated and that alterations in the expression level of InhA affect expression levels of KasA. Nonetheless, INH appeared to resemble TLM more closely in overall mode of action, and KasA levels appeared to be tightly correlated with INH sensitivity. C1 NIAID, TB Res Sect, Host Def Lab, Rockville, MD 20852 USA. RP Barry, CE (reprint author), NIAID, TB Res Sect, Host Def Lab, Twinbrook 2,Room 239,12441 Parklawn Dr, Rockville, MD 20852 USA. RI Barry, III, Clifton/H-3839-2012; Lee, Richard/J-4997-2013; Slayden, Richard/O-8626-2016 OI Lee, Richard/0000-0002-2397-0443; Slayden, Richard/0000-0001-6857-7277 FU Intramural NIH HHS [Z01 AI000783-11] NR 46 TC 107 Z9 111 U1 1 U2 11 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD NOV PY 2000 VL 38 IS 3 BP 514 EP 525 DI 10.1046/j.1365-2958.2000.02145.x PG 12 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 375MR UT WOS:000165404000009 PM 11069675 ER PT J AU Rodionov, DA Mironov, AA Rakhmaninova, AB Gelfand, MS AF Rodionov, DA Mironov, AA Rakhmaninova, AB Gelfand, MS TI Transcriptional regulation of transport and utilization systems for hexuronides, hexuronates and hexonates in gamma purple bacteria SO MOLECULAR MICROBIOLOGY LA English DT Article ID ESCHERICHIA-COLI K-12; COMPLETE GENOME SEQUENCE; ERWINIA-CHRYSANTHEMI; BINDING-SITES; GLUCONATE UPTAKE; GENE; CATABOLISM; PROTEINS; OPERON; PECTINOLYSIS AB The comparative approach is a powerful tool for the analysis of gene regulation in bacterial genomes. It can be applied to the analysis of regulons that have been studied experimentally as well as that of regulons for which no known regulatory sites are available. It is assumed that the set of co-regulated genes and the regulatory signal itself are conserved in related genomes, Here, we use genomic comparisons to study the regulation of transport and utilization systems for sugar acids in gamma purple bacteria Escherichia coli, Salmonella typhi, Klebsiella pneumoniae, Yersinia pestis, Erwinia chrysanthemi, Haemophilus influenzae and Vibrio cholerae, The variability of the operon structure and the location of the operator sites for the main transcription factors are demonstrated. The common metabolic map is combined with known and predicted regulatory interactions, It includes all known and predicted members of the GntR, UxuR/ExuR, KdgR, UidR and IdnR regulons, Moreover, most members of these regulons seem to be under catabolite repression mediated by CRP, The candidate UxuR/ExuR signal is proposed, the KdgR consensus is extended, and new operators for all transcription factors are identified in all studied genomes, Two new members of the KdgR regulon, a hypothetical ATP-dependent transport system OgtABCD and YjgK protein with unknown function, are detected. The former is likely to be the transport system for the products of pectin degradation, oligogalacturonides. C1 State Sci Ctr GosNIIGenetika, Moscow 113545, Russia. NIH, Natl Ctr Biotechnol Informat, Bethesda, MD 20205 USA. RP Rodionov, DA (reprint author), State Sci Ctr GosNIIGenetika, Moscow 113545, Russia. RI Mironov, Andrey/C-8024-2012; Gelfand, Mikhail/F-3425-2012 NR 35 TC 38 Z9 39 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD NOV PY 2000 VL 38 IS 4 BP 673 EP 683 DI 10.1046/j.1365-2958.2000.02115.x PG 11 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 383VM UT WOS:000165904800002 PM 11115104 ER PT J AU Zhou, YY Yang, D Zhu, WZ Zhang, SJ Wang, DJ Rohrer, DK Devic, E Kobilka, BK Lakatta, EG Cheng, HP Xiao, RP AF Zhou, YY Yang, D Zhu, WZ Zhang, SJ Wang, DJ Rohrer, DK Devic, E Kobilka, BK Lakatta, EG Cheng, HP Xiao, RP TI Spontaneous activation of beta 2-but not beta(1)-adrenoceptors expressed in cardiac myocytes from beta(1)beta(2) double knockout mice SO MOLECULAR PHARMACOLOGY LA English DT Article ID 3RD INTRACELLULAR LOOP; DOPAMINE D1B RECEPTOR; BETA(2)-ADRENERGIC RECEPTOR; TRANSGENIC MICE; HEART-FAILURE; CONSTITUTIVE ACTIVITY; VENTRICULAR MYOCYTES; INVERSE AGONISTS; CA2+ CURRENT; G-PROTEIN AB Although ligand-free, constitutive beta(2)-adrenergic receptor (AR) signaling has been demonstrated in naive cell lines and in transgenic mice overexpressing cardiac beta(2)-AR, it is unclear whether the dominant cardiac beta-AR subtype, beta(1)-AR, shares the ability of spontaneous activation. In the present study, we expressed human beta(1) -or beta(2)-AR via recombinant adenoviral infection in ventricular myocytes isolated from beta(1)beta(2)-AR double knockout mice, creating pure beta(1)-AR and beta(2)-AR systems with variable receptor densities. A contractile response to a nonselective beta-AR agonist, isoproterenol, was absent in double knockout mouse myocytes but was fully restored after adenoviral beta(1)-AR or adenoviral beta(2)-AR infection. Increasing the titer of adenoviral vectors (multiplicity of infection 10-1000) led to a dose-dependent expression of beta(1)- or beta(2)-AR with a maximal density of 1207 +/- 173 (36-fold over the wild-type control value) and 821 +/- 38 fmol/ mg protein (69-fold), respectively. Using confocal immunohistochemistry, we directly visualized the cellular distribution of beta(1)-AR and beta(2)-AR and found that both subtypes were distributed on the cell surface membrane and transverse tubules, resulting in a striated pattern. In the absence of ligand, beta(2)-AR expression resulted in graded increases in baseline cAMP and contractility up to 428% and 233% of control, respectively, at the maximal beta(2)-AR density. These effects were specifically reversed by a beta(2)-AR inverse agonist, ICI 118,551 (10(-7) M). In contrast, overexpression of beta(1)-AR, even at a greater density, failed to enhance either basal cAMP or contractility; the alleged beta(1)-AR inverse agonist, CGP 20712A (10(-6) M), had no significant effect on basal contraction in these cells. Thus, we conclude that acute beta(2)-AR overexpression in cardiac myocytes elicits significant physiological responses due to spontaneous receptor activation; however, this property is beta-AR subtype specific because beta(1)-AR does not exhibit agonist-independent spontaneous activation. C1 NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Peking Univ, Natl Lab Biomembrane & Membrane Biotechnol, Beijing, Peoples R China. Stanford Univ, Med Ctr, Howard Hughes Med Inst, Stanford, CA 94305 USA. RP Xiao, RP (reprint author), NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 37 TC 38 Z9 40 U1 1 U2 6 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 NOV PY 2000 VL 58 IS 5 BP 887 EP 894 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 364XG UT WOS:000089918900002 PM 11040034 ER PT J AU Koshimizu, TA Van Goor, F Tomic, M Wong, AOL Tanoue, A Tsujimoto, G Stojilkovic, SS AF Koshimizu, TA Van Goor, F Tomic, M Wong, AOL Tanoue, A Tsujimoto, G Stojilkovic, SS TI Characterization of calcium signaling by purinergic receptor-channels expressed in excitable cells SO MOLECULAR PHARMACOLOGY LA English DT Article ID GATED ION CHANNELS; P2X RECEPTORS; ATP RECEPTOR; DESENSITIZATION; ACTIVATION; NEURONS; CLONING; FAMILY; CDNA AB ATP-gated purinergic receptors (P2XRs) are a family of cationpermeable channels that conduct Ca(2+) and facilitate voltage-sensitive Ca(2+) entry in excitable cells. To study Ca(2+) signaling by P2XRs and its dependence on voltage-sensitive Ca(2+) influx, we expressed eight cloned P2XR subtypes individually in gonadotropin-releasing hormone-secreting neurons. In all cases, ATP evoked an inward current and a rise in [Ca(2+)](i). P2XR subtypes differed in the peak amplitude of [Ca(2+)](i) response independently of the level of receptor expression, with the following order: P2X(1)R, P2X(3)R, P2X(4)R, P2X(2b)R < P2X(2a)R, P2X(7)R. During prolonged agonist stimulation, Ca(2+) signals desensitized with different rates: P(2)X(3)R. P2X(1)R > P2X(2b)R > P2X(4)R >> P2X(2a)R >> P2X(7)R. The pattern of [Ca(2+)](i) response for each P2XR subtype was highly comparable with that of the depolarizing current, but the activation and desensitization rates were faster for the current than for [Ca(2+)](i). The P2X(1)R, P2X(3)R, and P2X(4)R-derived [Ca(2+)](i) signals were predominantly dependent on activation of voltage-sensitive Ca(2+) influx, both voltage-sensitive and -insensitive Ca(2+) entry pathways equally contributed to [Ca(2+)](i) responses in P2X(2a)R- and P2X(2b)R-expressing cells, and P2X(7)R operated as a nonselective pore capable of conducting larger amounts of Ca(2+) independently on the status of voltage-gated Ca(2+) channels. Thus, Ca(2+) signaling by homomeric P2XRs expressed in an excitable cell is subtype-specific, which provides an effective mechanism for generating variable [Ca(2+)](i) patterns in response to a common agonist. C1 NICHHD, Sect Cellular Signaling, ERRB, NIH, Bethesda, MD 20892 USA. Natl Childrens Med Res Ctr, Dept Mol Cellular Pharmacol, Tokyo, Japan. RP Stojilkovic, SS (reprint author), NICHHD, Sect Cellular Signaling, ERRB, NIH, Bldg 49,Room 6A-36,49 Convent Dr, Bethesda, MD 20892 USA. EM stankos@helix.nih.gov RI Wong, Anderson/A-2735-2010; Tomic, Melanija/C-3371-2016; OI Koshimizu, Taka-aki/0000-0001-5292-7535 NR 25 TC 53 Z9 55 U1 0 U2 4 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD NOV PY 2000 VL 58 IS 5 BP 936 EP 945 PG 10 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 364XG UT WOS:000089918900008 PM 11040040 ER PT J AU O'Keefe, BR Shenoy, SR Xie, D Zhang, WT Muschik, JM Currens, MJ Chaiken, I Boyd, MR AF O'Keefe, BR Shenoy, SR Xie, D Zhang, WT Muschik, JM Currens, MJ Chaiken, I Boyd, MR TI Analysis of the interaction between the HIV-Inactivating protein cyanovirin-N and soluble forms of the envelope glycoproteins gp120 and gp41 SO MOLECULAR PHARMACOLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; OLIGOSACCHARIDE STRUCTURES; FUSION MECHANISM; PEPTIDE; INHIBITION; BINDING; CYANOBACTERIUM; INFECTIVITY; DISCOVERY; ANTIBODY AB The novel virucidal protein cyanovirin-N (CV-N) binds with equally high affinity to soluble forms of either H9 cell-produced or recombinant glycosylated HIV-1 gp120 (sgp120) or gp160 (sgp160). Fluorescence polarization studies showed that CV-N is also capable of binding to the glycosylated ectodomain of the HIV-envelope protein gp41 (sgp41) (as well as SIV glycoprotein 32), albeit with considerably lower affinity than the sgp120/CV-N interaction. Pretreatment of CV-N with either sgp120 or sgp41 abrogated the neutralizing activity of CV-N against intact, infectious HIV-1 virions. Isothermal calorimetry and optical biosensor binding studies showed that CV-N bound to recombinant sgp120 with a K-d value ranging from 2 to 45 nM and to sgp41 with a K-d value of 606 nM; furthermore, they indicated an approximate 5:1 stoichiometry for CV-N binding to sgp120 and a 1:1 stoichiometry for CV-N binding to sgp41. Circular dichroism studies additionally illuminated the binding of CV-N with both sgp120 and sgp41, providing the first direct evidence that conformational changes are a consequence of CV-N interactions with both HIV-1 envelope glycoproteins. C1 Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag, Dev Therapeut Program, Lab Drug Discovery Res & Dev,SAIC Frederick, Frederick, MD 21702 USA. Frederick Canc Res & Dev Ctr, SAIC Frederick, Struct Biochem Program, Frederick, MD 21702 USA. Univ Penn, Sch Med, Dept Med, Philadelphia, PA 19104 USA. RP Boyd, MR (reprint author), Frederick Canc Res & Dev Ctr, Div Canc Treatment & Diag, Dev Therapeut Program, Lab Drug Discovery Res & Dev,SAIC Frederick, Bldg 1052,Rm 121, Frederick, MD 21702 USA. FU NIGMS NIH HHS [P01-GM56550-01] NR 32 TC 59 Z9 65 U1 0 U2 4 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 NOV PY 2000 VL 58 IS 5 BP 982 EP 992 PG 11 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 364XG UT WOS:000089918900013 PM 11040045 ER PT J AU Olivares-Reyes, JA Jayadev, S Hunyady, L Catt, KJ Smith, RD AF Olivares-Reyes, JA Jayadev, S Hunyady, L Catt, KJ Smith, RD TI Homologous and heterologous phosphorylation of the AT(2) angiotensin receptor by protein kinase C SO MOLECULAR PHARMACOLOGY LA English DT Article ID AGONIST-INDUCED INTERNALIZATION; II TYPE-2 RECEPTORS; COUPLED RECEPTOR; TYROSINE PHOSPHATASE; MOLECULAR-CLONING; AT(1A) RECEPTOR; AT2 RECEPTORS; CELLS; IDENTIFICATION; INHIBITION AB The angiotensin AT(2) receptor is an atypical seven transmembrane domain receptor that is coupled to activation of tyrosine phosphatase and inhibition of MAP kinase, and does not undergo agonist-induced internalization. An investigation of the occurrence and nature of AT(2) receptor phosphorylation revealed that phorbol ester-induced activation of protein kinase C (PKC) in HA-AT(2) receptor-expressing COS-7 cells caused rapid and specific phosphorylation of a single residue (Ser(354))located in the cytoplasmic tail of the receptor. Agonist activation of AT(2) receptors by angiotensin II (Ang II) also caused rapid PKC-dependent phosphorylation of Ser(354) that was prevented by the AT(2) antagonist, PD123177, and by inhibitors of PKC. In cells coexpressing AT(1) and AT(2) receptors, Ang II-induced phosphorylation of the AT(2) receptor was reduced by either PD123177 or the AT(1) receptor antagonist, DuP753, and was abolished by treatment with both antagonists or with PKC inhibitors. These findings indicate that the AT(2) receptor is rapidly phosphorylated via PKC during homologous activation by Ang II, and also undergoes heterologous PKC-dependent phosphorylation during activation of the AT(1) receptor. The latter process may regulate the counteracting effects of AT(2) receptors on growth responses to AT(1) receptor activation. C1 NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Semmelweis Univ, Dept Physiol, H-1085 Budapest, Hungary. RP NICHHD, Endocrinol & Reprod Res Branch, NIH, Bldg 49,Rm 6A-36,49 Convent Dr, Bethesda, MD 20892 USA. EM catt@helix.nih.gov FU PHS HHS [979004] NR 38 TC 11 Z9 11 U1 0 U2 0 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0026-895X EI 1521-0111 J9 MOL PHARMACOL JI Mol. Pharmacol. PD NOV PY 2000 VL 58 IS 5 BP 1156 EP 1161 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 364XG UT WOS:000089918900033 PM 11040065 ER PT J AU Manji, HK Moore, GJ Rajkowska, G Chen, G AF Manji, HK Moore, GJ Rajkowska, G Chen, G TI Neuroplasticity and cellular resilience in mood disorders SO MOLECULAR PSYCHIATRY LA English DT Review DE atrophy; cell death; neuroprotection; lithium; bcl-2; MAP kinase; neurite; neurotrophic; neurogenesis; N-acetylaspartate; gray matter ID MAGNETIC-RESONANCE SPECTROSCOPY; GLYCOGEN-SYNTHASE KINASE-3; SUBGENUAL PREFRONTAL CORTEX; REDUCES TAU-PHOSPHORYLATION; BRAIN PHOSPHORUS-METABOLISM; FAMILY TRANSCRIPTION FACTOR; RECURRENT MAJOR DEPRESSION; CHRONIC LITHIUM TREATMENT; CEREBELLAR GRANULE CELLS; TRKB MESSENGER-RNA AB Although mood disorders have traditionally been regarded as good prognosis diseases, a growing body of data suggests that the long-term outcome for many patients is often much less favorable than previously thought, Recent morphometric studies have been investigating potential structural brain changes in mood disorders, and there is now evidence from a variety of sources demonstrating significant reductions in regional CNS volume, as well as regional reductions in the numbers and/or sizes of glia and neurons. Furthermore, results from recent clinical and preclinical studies investigating the molecular and cellular targets of mood stabilizers and antidepressants suggest that a reconceptualization about the pathophysiology and optimal long-term treatment of recurrent mood disorders may be warranted. It is proposed that impairments of neuroplasticity and cellular resilience may underlie the pathophysiology of mood disorders, and further that optimal long-term treatment for these severe illnesses may only be achieved by the early and aggressive use of agents with neurotrophic/neuroprotective effects, It is noteworthy that lithium, valproate and antidepressants indirectly regulate a number of factors involved in cell survival pathways including CREB, BDNF, bcl-2 and MAP kinases, and may thus bring about some of their delayed long-term beneficial effects via underappreciated neurotrophic effects. The development of novel treatments which more directly target molecules involved in critical CNS cell survival and cell death pathways have the potential to enhance neuroplasticity and cellular resilience, and thereby modulate the long-term course and trajectory of these devastating illnesses. C1 NIMH, Mol Pathophysiol Lab, Bethesda, MD 20892 USA. Wayne State Univ, Sch Med, Mol Pathophysiol Lab, Detroit, MI 48202 USA. Wayne State Univ, Sch Med, Cellular & Clin Neurobiol Program, Dept Psychiat & Behav Neurosci, Detroit, MI 48202 USA. Wayne State Univ, Sch Med, Dept Pharmacol, Detroit, MI 48201 USA. Wayne State Univ, Sch Med, Dept Radiol, Detroit, MI USA. Univ Mississippi, Sch Med, Dept Psychiat & Human Behav, University, MS 38677 USA. RP Manji, HK (reprint author), NIMH, Mol Pathophysiol Lab, 10 Ctr Dr,10-4N-222 MSC 1381, Bethesda, MD 20892 USA. RI Moore, Gregory/E-7184-2010; Chen, Guang/A-2570-2017 OI Moore, Gregory/0000-0001-8541-3194; FU NIMH NIH HHS [MH57743, MH59107]; PHS HHS [55872] NR 163 TC 223 Z9 230 U1 4 U2 12 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1359-4184 J9 MOL PSYCHIATR JI Mol. Psychiatr. PD NOV PY 2000 VL 5 IS 6 BP 578 EP 593 DI 10.1038/sj.mp.4000811 PG 16 WC Biochemistry & Molecular Biology; Neurosciences; Psychiatry SC Biochemistry & Molecular Biology; Neurosciences & Neurology; Psychiatry GA 372MC UT WOS:000165236600005 PM 11126389 ER PT J AU Jabbari, B Molloy, FM Erickson, M Floeter, MK AF Jabbari, B Molloy, FM Erickson, M Floeter, MK TI Bilateral painful hand-moving fingers: Electrophysiological assessment of the central nervous system oscillator SO MOVEMENT DISORDERS LA English DT Article DE painful hand-moving fingers; painful leg-moving toes; EMG ID LEGS; TOES; INHIBITION AB We describe a 35-year-old woman who presented with the syndrome of painful hand-moving fingers on the right side. Eight months later, she developed similar finger movements and hand discomfort on the left side. She had a history of hand trauma and recurrent shoulder dislocation on the right side. Kinesiologic electromyography suggested a common central oscillator for finger movements in both hands. Electrophysiological assessment of spinal alpha motor neuron excitability, reciprocal inhibition, and Renshaw cell inhibition failed to show any abnormalities. Somatosensory evoked potential test showed marked attenuation of N20 potential recorded from the left somatosensory cortex; paired transcortical magnetic stimulation of the left motor cortex suggested failure of cortical facilitation. The data suggest that the central oscillator responsible for finger movements is located above the spinal cord level in this patient. C1 Uniformed Serv Univ Hlth Sci, Dept Neurol, Bethesda, MD 20814 USA. NINDS, EMG Sect, NIH, Bethesda, MD 20892 USA. RP Jabbari, B (reprint author), Uniformed Serv Univ Hlth Sci, Dept Neurol, 4301 Jones Bridge Rd, Bethesda, MD 20814 USA. NR 15 TC 8 Z9 8 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0885-3185 J9 MOVEMENT DISORD JI Mov. Disord. PD NOV PY 2000 VL 15 IS 6 BP 1259 EP 1263 DI 10.1002/1531-8257(200011)15:6<1259::AID-MDS1032>3.0.CO;2-5 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 372MD UT WOS:000165236700032 PM 11104217 ER PT J AU Aragon-Alcaide, L Strunnikov, AV AF Aragon-Alcaide, L Strunnikov, AV TI Functional dissection of in vivo interchromosome association in Saccharomyces cerevisiae SO NATURE CELL BIOLOGY LA English DT Article ID MEIOTIC PROPHASE; MITOTIC RECOMBINATION; BUDDING YEAST; NUCLEAR-ORGANIZATION; GENE-EXPRESSION; DNA; CENTROMERE; PROTEINS; CELLS; TRANSVECTION AB Homologue pairing mediates both recombination and segregation of chromosomes at meiosis I. The recognition of nucleic-acid-sequence homology within the somatic nucleus has an impact on DNA repair and epigenetic control of gene expression. Here we investigate interchromosomal interactions using a non-invasive technique that allows tagging and visualization of DNA sequences in vegetative and meiotic live yeast cells. In non-meiotic cells, chromosomes are ordered in the nucleus, but preferential pairing between homologues is not observed. Association of tagged chromosomal domains occurs irrespective of their genomic location, with some preference for similar chromosomal positions. Here we describe a new phenomenon that promotes associations between sequence-identical ectopic tags with a tandem-repeat structure. These associations, termed interchromosome trans-associations, may underlie epigenetic phenomena. C1 NICHD, Unit Chromosome Struct & Funct, NIH, Bethesda, MD 20892 USA. RP Aragon-Alcaide, L (reprint author), NICHD, Unit Chromosome Struct & Funct, NIH, 18T Lib Dr, Bethesda, MD 20892 USA. OI Strunnikov, Alexander/0000-0002-9058-2256 FU Intramural NIH HHS [Z99 AI999999, Z01 HD001903-11] NR 41 TC 37 Z9 37 U1 0 U2 1 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 1465-7392 J9 NAT CELL BIOL JI Nat. Cell Biol. PD NOV PY 2000 VL 2 IS 11 BP 812 EP 818 PG 7 WC Cell Biology SC Cell Biology GA 371YU UT WOS:000165207500015 PM 11056536 ER PT J AU Tong, ZB Gold, L Pfeifer, KE Dorward, H Lee, E Bondy, CA Dean, J Nelson, LM AF Tong, ZB Gold, L Pfeifer, KE Dorward, H Lee, E Bondy, CA Dean, J Nelson, LM TI Mater, a maternal effect gene required for early embryonic development in mice SO NATURE GENETICS LA English DT Article ID MOUSE EMBRYO; TRANSCRIPTION; ACTIVATION; PROTEINS; EXPRESSION; TRANSITION; DIVERSITY; FAMILY; STAGE C1 NIDDK, Cellular & Dev Biol Lab, NIH, Bethesda, MD USA. NICHD, Dev Endocrinol Branch, NIH, Bethesda, MD USA. NICHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD USA. Georgetown Univ, Med Ctr, Dept Pediat, Washington, DC 20007 USA. RP Dean, J (reprint author), NIDDK, Cellular & Dev Biol Lab, NIH, Bethesda, MD USA. OI Pfeifer, Karl/0000-0002-0254-682X NR 15 TC 317 Z9 337 U1 1 U2 6 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD NOV PY 2000 VL 26 IS 3 BP 267 EP 268 PG 2 WC Genetics & Heredity SC Genetics & Heredity GA 371JU UT WOS:000165176500008 PM 11062459 ER PT J AU Bamford, RN Roessler, E Burdine, RD Saplakoglu, U dela Cruz, J Splitt, M Towbin, J Bowers, P Marino, B Schier, AF Shen, MM Muenke, M Casey, B AF Bamford, RN Roessler, E Burdine, RD Saplakoglu, U dela Cruz, J Splitt, M Towbin, J Bowers, P Marino, B Schier, AF Shen, MM Muenke, M Casey, B TI Loss-of-function mutations in the EGF-CFC gene CFC1 are associated with human left-right laterality defects SO NATURE GENETICS LA English DT Article ID LEFT-RIGHT AXIS; ONE-EYED PINHEAD; GASTRULATION; ZEBRAFISH; MALFORMATIONS; MESODERM AB Ail vertebrates display a characteristic asymmetry of internal organs with the cardiac apex, stomach and spleen towards the left, and the liver and gall bladder on the right(1-3). Left-right (L-R) axis abnormalities or laterality defects are common in humans (1 in 8,500 live births). Several genes (such as Nodal, Ebaf and Pitx2) have been implicated in L-R organ positioning in model organisms(2-4), In humans, relatively few genes have been associated with a small percentage of human situs defects. These include ZIC3 (ref. 5), LEFTB (formerly LEFTY2; ref. 6) and ACVR2B (encoding activin receptor IIB; ref, 7). The EGF-CFC genes(8), mouse Cfc1 (encoding the Cryptic protein; ref. 9) and zebrafish one-eyed pinhead (oep; refs 10,11) are essential for the establishment of the L-R axis(12,13). EGF-CFC proteins act as co-factors for Nodal-related signals(11), which have also been implicated in L-R axis development(4). Here we identify loss-of-function mutations in human CFC1 (encoding the CRYPTIC protein) in patients with heterotaxic phenotypes (randomized organ positioning). The mutant proteins have aberrant cellular localization in transfected cells and are functionally defective in a zebrafish oepmutant rescue assay. Our findings indicate that the essential role of EGF-CFC genes and Nodal signalling in left-right axis formation is conserved from fish to humans. Moreover, our results support a role for environmental and/or genetic modifiers in determining the ultimate phenotype in humans. C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. NYU, Sch Med, Dept Cell Biol, Skirball Inst Biomol Med,Dev Genet Program, New York, NY 10016 USA. Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Ctr Adv Biotechnol & Med, Piscataway, NJ 08854 USA. Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Pediat, Piscataway, NJ 08854 USA. Univ Newcastle Upon Tyne, Dept Human Genet, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England. Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. Yale Univ, Sch Med, Dept Pediat, New Haven, CT 06510 USA. Bambino Gesu Childrens Hosp, Dept Cardiol, Rome, Italy. Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA. RP Muenke, M (reprint author), NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. RI Shen, Michael/C-7546-2009; OI Shen, Michael/0000-0002-4042-1657; Burdine, Rebecca/0000-0001-6620-5015 NR 22 TC 206 Z9 216 U1 1 U2 13 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 NOV PY 2000 VL 26 IS 3 BP 365 EP 369 PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 371JU UT WOS:000165176500031 PM 11062482 ER PT J AU Di Prospero, NA Tagle, DA AF Di Prospero, NA Tagle, DA TI Normal and mutant huntingtin: Partners in crime? SO NATURE MEDICINE LA English DT Editorial Material ID NEURONAL INTRANUCLEAR INCLUSIONS; DISEASE C1 NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. RP Di Prospero, NA (reprint author), NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. NR 12 TC 3 Z9 3 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 NOV PY 2000 VL 6 IS 11 BP 1208 EP 1209 PG 2 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 370GH UT WOS:000165114800015 PM 11062523 ER EF