122 results on '"Costa, R. H."'
Search Results
2. An N-terminal inhibitory domain modulates activity of FoxM1 during cell cycle
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Park, H J, Wang, Z, Costa, R H, Tyner, A, Lau, L F, and Raychaudhuri, P
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- 2008
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3. REDUCTION OF EXCESS SLUDGE PRODUCTION IN AN ACTIVATED SLUDGE SYSTEM BASED ON LYSIS-CRYPTIC GROWTH, UNCOUPLING METABOLISM AND FOLIC ACID ADDITION
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Velho, V. F., primary, Daudt, G. C., additional, Martins, C. L., additional, Belli Filho, P., additional, and Costa, R. H. R., additional
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- 2016
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4. Simulation and calibration of a full-scale sequencing batch reactor for wastewater treatment
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Oselame, M. C., primary, Fernandes, H., additional, and Costa, R. H. R., additional
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- 2014
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5. Sequencing batch reactor operation for treating wastewater with aerobic granular sludge
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Jungles, M. K., primary, Campos, J. L., additional, and Costa, R. H. R., additional
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- 2014
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6. Cytokine regulation of the liver transcription factor hepatocyte nuclear factor-3β is mediated by the C/EBP family and interferon regulatory factor 1
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Samadani, U., Porcella, A., Pani, L., Peter Johnson, Burch, J. B. E., Pine, R., and Costa, R. H.
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- 1995
7. An N-terminal inhibitory domain modulates activity of FoxM1 during cell cycle
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Park, H J, primary, Wang, Z, additional, Costa, R H, additional, Tyner, A, additional, Lau, L F, additional, and Raychaudhuri, P, additional
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- 2007
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8. Hepatocyte nuclear factor 3/fork head homolog 11 is expressed in proliferating epithelial and mesenchymal cells of embryonic and adult tissues
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Ye, H, primary, Kelly, T F, additional, Samadani, U, additional, Lim, L, additional, Rubio, S, additional, Overdier, D G, additional, Roebuck, K A, additional, and Costa, R H, additional
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- 1997
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9. Thyroid transcription factor-1, hepatocyte nuclear factor-3beta, surfactant protein B, C, and Clara cell secretory protein in developing mouse lung.
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Zhou, L, primary, Lim, L, additional, Costa, R H, additional, and Whitsett, J A, additional
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- 1996
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10. Decreased expression of hepatocyte nuclear factor 3 alpha during the acute-phase response influences transthyretin gene transcription
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Qian, X, primary, Samadani, U, additional, Porcella, A, additional, and Costa, R H, additional
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- 1995
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11. The DNA-binding specificity of the hepatocyte nuclear factor 3/forkhead domain is influenced by amino-acid residues adjacent to the recognition helix
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Overdier, D G, primary, Porcella, A, additional, and Costa, R H, additional
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- 1994
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12. Performance and Kinectics Aspects of Nitrogen Removal in a Biofilm Sequencing Batch Reactor.
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Costa, R. H. R., Wolff, D. B., and Souto, V. S.
- Abstract
A biofilm sequencing batch reactor with a volume of 1.42 m
3 , nylon nets providing a 4,140 m2 /m3 support area for biofilms and an automated operation with 8 hour cycles was studied. The duration of the experiment was 135 days. Removal efficiencies e≧ 80% were obtained for carbonaceous matter, producing an effluent with 31±26.8 mg/L of filtered COD, 7±3.6 mg/L of BOD5 and 12±3.2 mg/L of TOC. The average removal efficiency of ammonium was 77 ± 16.6%, with a mean concentration in the effluent of 14 ± 10.2 mg NH4 -N/L. The denitrification efficiency was 80±14.7%. The effluent characteristics met the requirements of Brazilian environmental standard for discharge to receiving water bodies. A kinetic study of nitrification and denitrification showed that during the aerobic phase the specific rate of ammonium consumption was 0.057 g NH4 -N/g VSS.d and the production of NOx-N was 0.074 g NOx-N/g VSS.d, while the specific rate of NOx-N consumption was 0.05 g NOx-N/g VSS.d during the anoxic phase. The suspended and fixed biomass was composed of 50% ammonium-oxidizing bacteria (AOB). [ABSTRACT FROM AUTHOR]- Published
- 2013
13. Identification of nine tissue-specific transcription factors of the hepatocyte nuclear factor 3/forkhead DNA-binding-domain family.
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Clevidence, D E, primary, Overdier, D G, additional, Tao, W, additional, Qian, X, additional, Pani, L, additional, Lai, E, additional, and Costa, R H, additional
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- 1993
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14. Hepatocyte nuclear factor 3 beta contains two transcriptional activation domains, one of which is novel and conserved with the Drosophila fork head protein.
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Pani, L, primary, Overdier, D G, additional, Porcella, A, additional, Qian, X, additional, Lai, E, additional, and Costa, R H, additional
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- 1992
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15. The restricted promoter activity of the liver transcription factor hepatocyte nuclear factor 3 beta involves a cell-specific factor and positive autoactivation.
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Pani, L, primary, Quian, X B, additional, Clevidence, D, additional, and Costa, R H, additional
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- 1992
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16. Similarities in transthyretin gene expression and differences in transcription factors: liver and yolk sac compared to choroid plexus.
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Costa, R H, primary, Van Dyke, T A, additional, Yan, C, additional, Kuo, F, additional, and Darnell, J E, additional
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- 1990
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17. HNF-3A, a hepatocyte-enriched transcription factor of novel structure is regulated transcriptionally.
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Lai, E, primary, Prezioso, V R, additional, Smith, E, additional, Litvin, O, additional, Costa, R H, additional, and Darnell, J E, additional
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- 1990
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18. Multiple Hepatocyte-Enriched Nuclear Factors Function in the Regulation of Transthyretin and α1-Antitrypsin Genes
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Costa, R H, Grayson, D R, and Darnell, J E
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Binding Sites ,Base Sequence ,Nuclear Proteins ,nutritional and metabolic diseases ,DNA ,Cell Biology ,DNA-Binding Proteins ,Mice ,Enhancer Elements, Genetic ,Gene Expression Regulation ,Liver ,alpha 1-Antitrypsin ,Animals ,Humans ,Prealbumin ,Molecular Biology ,Research Article - Abstract
Transthyretin (TTR) and alpha 1-antitrypsin (alpha 1-AT) are expressed at high levels in the liver and also in at least one other cell type. We report here a detailed analysis of the proximal regulatory region of the TTR gene, which has uncovered two new DNA-binding factors that are present mainly (or only) in hepatocytes. One of these new factors, hepatocyte nuclear factor 3 (HNF-3), binds to two sites that are crucial in TTR expression as well as to two additional sites in the alpha 1-AT proximal enhancer region. The second new factor, HNF-4, binds to two sites in TTR that are required for gene activity. We had previously identified binding sites for another hepatocyte-enriched DNA-binding protein (C/EBP or a relative thereof), and additional promoter-proximal sites for that protein in both TTR and alpha 1-AT are also reported here. From these results it seems clear that cell-specific expression is not simply the result of a single cell-specific factor for each gene but the result of a combination of such factors. The variation and distribution of such factors among different cell types could be an important basis for the coordinate expression of the TTR and alpha 1-AT genes in the liver or the discordant transcriptional activation of these genes in a few other cell types. The identification of such cell-enriched factors is a necessary prelude to understanding the basis for cell specificity.
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- 1989
19. Persistent expression of HNF6 in islet endocrine cells causes disrupted islet architecture and loss of beta cell function.
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Gannon, M, Ray, M K, Van Zee, K, Rausa, F, Costa, R H, and Wright, C V
- Abstract
We used transgenesis to explore the requirement for downregulation of hepatocyte nuclear factor 6 (HNF6) expression in the assembly, differentiation, and function of pancreatic islets. In vivo, HNF6 expression becomes downregulated in pancreatic endocrine cells at 18. 5 days post coitum (d.p.c.), when definitive islets first begin to organize. We used an islet-specific regulatory element (pdx1(PB)) from pancreatic/duodenal homeobox (pdx1) gene to maintain HNF6 expression in endocrine cells beyond 18.5 d.p.c. Transgenic animals were diabetic. HNF6-overexpressing islets were hyperplastic and remained very close to the pancreatic ducts. Strikingly, alpha, delta, and PP cells were increased in number and abnormally intermingled with islet beta cells. Although several mature beta cell markers were expressed in beta cells of transgenic islets, the glucose transporter GLUT2 was absent or severely reduced. As glucose uptake/metabolism is essential for insulin secretion, decreased GLUT2 may contribute to the etiology of diabetes in pdx1(PB)-HNF6 transgenics. Concordantly, blood insulin was not raised by glucose challenge, suggesting profound beta cell dysfunction. Thus, we have shown that HNF6 downregulation during islet ontogeny is critical to normal pancreas formation and function: continued expression impairs the clustering of endocrine cells and their separation from the ductal epithelium, disrupts the spatial organization of endocrine cell types within the islet, and severely compromises beta cell physiology, leading to overt diabetes.
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- 2000
20. A cell-specific enhancer of the mouse alpha 1-antitrypsin gene has multiple functional regions and corresponding protein-binding sites
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Grayson, D R, Costa, R H, Xanthopoulos, K G, and Darnell, J E
- Abstract
We have previously described the isolation and characterization of genomic clones corresponding to the mouse alpha 1-antitrypsin gene (Krauter et al., DNA 5:29-36, 1986). In this report, we have analyzed the DNA sequences upstream of the RNA start site that direct hepatoma cell-specific expression of this gene when incorporated into recombinant plasmids. The 160 nucleotides 5' to the cap site direct low-level expression in hepatoma cells, and sequences between -520 and -160 bp upstream of the RNA start site functioned as a cell-specific enhancer of expression both with the alpha 1-antitrypsin promoter and when combined with a functional beta-globin promoter. Within the enhancer region, three binding sites for proteins present in hepatoma nuclear extracts were identified. The location of each site was positioned, using both methylation protection and methylation interference experiments. Each protein-binding site correlated with a functionally important region necessary for full enhancer activity. These experiments demonstrated a complex arrangement of regulatory elements comprising the alpha 1-antitrypsin enhancer. Significant qualitative differences exist between the findings presented here and the cis-acting elements operative in regulating expression of the human alpha 1-antitrypsin gene (Ciliberto et al., Cell 41:531-540, 1985; De Simone et al., EMBO J. 6:2759-2766, 1987).
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- 1988
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21. Transcriptional control of the mouse prealbumin (transthyretin) gene: both promoter sequences and a distinct enhancer are cell specific
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Costa, R H, Lai, E, and Darnell, J E
- Abstract
The mouse genomic clone for the prealbumin (transthyretin) gene was cloned, and its upstream regulatory regions were analyzed. The 200 nucleotides 5' to the cap site when placed within a recombinant plasmid were sufficient to direct transient expression in HepG2 (human hepatoma) cells, but this DNA region did not support expression in HeLa cells. The sequence of the 200-nucleotide region is highly conserved between mouse and human DNA and can be considered a cell-specific promoter. Deletions of this promoter region identified a crucial element for cell-specific expression between 151 and 110 nucleotides 5' to the RNA start site. A region situated at about 1.6 to 2.15 kilobases upstream of the RNA start site was found to stimulate expression 10-fold in HepG2 cells but not in HeLa cells. This far upstream element was invertible and increased expression from the beta-globin promoter in HepG2 cells. Unlike the simian virus 40 enhancer, the prealbumin enhancer would not stimulate beta-globin synthesis in HeLa cells, and even the simian virus 40 enhancer did not stimulate the prealbumin promoter in HeLa cells. Thus, we identified in the prealbumin gene two DNA elements that respond in a cell-specific manner: a proximal promoter including a crucial sequence between -108 and -151 nucleotides and a distant enhancer element located between 1.6 and 2.15 kilobases upstream.
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- 1986
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22. The winged helix transcriptional activator HFH-8 is expressed in the mesoderm of the primitive streak stage of mouse embryos and its cellular derivatives
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Peterson, R. S., Lim, L., Ye, H., Zhou, H., Overdier, D. G., and Costa, R. H.
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- 1997
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23. The winged helix transcriptional activator HFH-3 is expressed in the distal tubules of embryonic and adult mouse kidney.
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Overdier, D G, Ye, H, Peterson, R S, Clevidence, D E, and Costa, R H
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The hepatocyte nuclear factor-3 (HNF-3)/fork head homolog (HFH) proteins are an extensive family of transcription factors, which share homology in the winged helix DNA binding domain. Members of the HFH/winged helix family have been implicated in cell fate determination during pattern formation, in organogenesis, and in cell-type-specific gene expression. In this study we isolated a full-length HFH-3 cDNA clone from a human kidney library which encoded a 351-amino acid protein containing a centrally located winged helix DNA binding domain. We demonstrate that HFH-3 is a potent transcriptional activator requiring 138 C-terminal residues for activity. We used in situ hybridization to demonstrate that HFH-3 expression is restricted to the epithelium of the renal distal convoluted tubules. We determined the HFH-3 DNA binding consensus sequence by in vitro DNA binding site selection using recombinant HFH-3 protein and used this consensus sequence to identify putative HFH-3 target genes expressed there. These putative HFH-3 target genes include the Na/K-ATPase, Na/H and anion exchangers, E-cadherin, and mineralocorticoid receptor genes as well as genes for the transcription factors HNF-1, vHNF-1, and HNF-4.
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- 1997
24. Differential regulation of interleukin-8 and intercellular adhesion molecule-1 by H2O2 and tumor necrosis factor-alpha in endothelial and epithelial cells.
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Lakshminarayanan, V, Beno, D W, Costa, R H, and Roebuck, K A
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The reactive oxygen intermediate H2O2 can function as a signaling molecule to activate gene expression. In this study, we demonstrate that oxidant stress induced by tumor necrosis factor alpha (TNFalpha) or H2O2 differentially regulates intercellular adhesion molecule-1 (ICAM-1) and interleukin-8 (IL-8) gene expression in endothelial and epithelial cells. Northern blot analysis revealed that TNFalpha induced both ICAM-1 and IL-8 expression in either the A549 lung epithelial cell line or the human microvessel endothelial cell line (HMEC-1). In contrast, H2O2 selectively induced only ICAM-1 in HMEC-1 and only IL-8 in A549. This cell type-specific pattern of IL-8 expression was also observed in several other endothelial and epithelial cells. TNFalpha induced greater IL-8 gene expression as compared with H2O2, but the kinetics of induction were similar. The induction of epithelial IL-8 message was accompanied by a corresponding increase in functional IL-8 protein secretion as determined by a neutrophil motility assay. The increased neutrophil motility stimulated by conditioned media from H2O2- or TNFalpha-exposed A549 cells was completely inhibited by an anti-IL-8 antibody. TNFalpha and H2O2 also induced a differential pattern of CC chemokine expression in A549. While TNFalpha induced both RANTES and MCP-1, H2O2 induced only MCP-1. These data suggest that epithelial cells under oxidant stress contribute to the inflammatory cytokine network by selective production of IL-8, MCP-1, and RANTES, which may critically influence the site-specific recruitment of leukocyte subsets.
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- 1997
25. Characterization of a major late herpes simplex virus type 1 mRNA
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Costa, R H, Devi, B G, Anderson, K P, Gaylord, B H, and Wagner, E K
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A major, late 6-kilobase (6-kb) mRNa mapping in the large unique region of herpes simplex virus type 1 (HSV-1) was characterized by using two recombinant DNA clones, one containing EcoRI fragment G (0.190 to 0.30 map units) in lambda. WES.B (L. Enquist, M. Madden, P. Schiop-Stansly, and G. Vandl Woude, Science 203:541-544, 1979) and one containing HindIII fragment J (0.181 to 0.259 map units) in pBR322. This 6-kb mRNA had its 3' end to the left of 0.231 on the prototypical arrangement of the HSV-1 genome and was transcribed from right to left. It was bounded on both sides by regions containing a large number of distinct mRNA species, and its 3' end was partially colinear with a 1.5-kb mRNA which encoded a 35,000-dalton polypeptide. The 6-kb mRNA encoded a 155,000-dalton polypeptide which was shown to be the only one of this size detectable by hybrid-arrested translation encoded by late polyadenylated polyribosomal RNA. The S1 nuclease mapping experiments indicated that there were no introns in the coding sequence for this mRNA and that its 3' end mapped approximately 800 nucleotides to the left of the BglII site at 0.231, whereas its 5' end extended very close to the BamHI site at 0.266.
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- 1981
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26. Detailed characterization of the mRNA mapping in the HindIII fragment K region of the herpes simplex virus type 1 genome
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Anderson, K P, Frink, R J, Devi, G B, Gaylord, B H, Costa, R H, and Wagner, E K
- Abstract
We have isolated as recombinant DNA clones, in the plasmid pBR322, regions of the herpesvirus type 1 genome spanning the region between 0.53 and 0.6 on the prototypical arrangement. This 11,000-base-pair region corresponds to 10% of the large unique region and encodes five major and several minor mRNA species abundant at different times after infection, which range in length from 7 to 1 kilobase. In this report, we have used RNA transfer blots and S1 nuclease digestion of hybrids between viral DNA and polyribosomal RNA to precisely localize (+/- 0.1 kilobase) these mRNA's. Comparison of neutral and alkaline gels of S1 nuclease-digested hybrids indicates no internal introns in the coding sequences of these mRNA's, although noncontiguous leader sequences near (ca. 0.1 kilobase) the 5' ends of any or all mRNA's could not be excluded. The 5' ends of several late mRNA's that are encoded opposite DNA strands map very close to one another, and the 3' ends of a major late and a major early mRNA, which are partially colinear, terminate in the same region. In vitro translation of the viral mRNA's isolated by hybridization with DNA bound to cellulose and fractionation of mRNA species on denaturing agarose gels allowed us to assign specific polypeptide products to each of the mRNA's characterized. Among other results, it was demonstrated unequivocally that two major late mRNA's, which partially overlap, encode the same polypeptide.
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- 1981
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27. Virus-induced modification of the host cell is required for expression of the bacterial chloramphenicol acetyltransferase gene controlled by a late herpes simplex virus promoter (VP5)
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Costa, R H, Draper, K G, Devi-Rao, G, Thompson, R L, and Wagner, E K
- Abstract
The requirements for expression of genes under the control of early (alkaline exonuclease) and late (VP5) herpes simplex virus type 1 (HSV-1) gene promoters were examined in a transient expression assay, using the bacterial chloramphenicol acetyltransferase gene as an expression marker. Both promoters were induced, resulting in the production of high levels of the enzyme upon low-multiplicity infection by HSV-1. S1 nuclease analysis of hybrids between RNA isolated from infected cells containing HSV-1 promoter constructs and marker gene DNA demonstrated normal transcriptional initiation of the marker gene directed by the viral promoters. Viral DNA sequences no more than 125 bases 5' of the putative transcriptional cap site were sufficient for maximum activity of the late promoter. In contrast to expression controlled by the early gene, the late promoter was not active at a measurable level in uninfected cells until DNA sequences between 75 and 125 bases 5' of the transcriptional cap site were deleted. Cotransfection of cells with the expression marker controlled by HSV promoters and a cosmid containing HSV alpha (immediate-early) genes indicated that full expression of both early and late promoters requires the same virus-induced host cell modifications. Inhibition of viral DNA synthesis results in an increased rate of transient expression of marker genes under control of either early or late promoters in contrast to the situation in normal virus infection. These data provide evidence that the normal course of expression of late HSV genes involves negative modulation of potentially active promoters in the infected cell.
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- 1985
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28. Molecular basis of the glycoprotein-C-negative phenotype of herpes simplex virus type 1 macroplaque strain
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Draper, K G, Costa, R H, Lee, G T, Spear, P G, and Wagner, E K
- Abstract
The basis for the inability of the macroplaque (MP) strain of herpes simplex virus type 1 to express mature glycoprotein C (gC) was examined. RNA transfer (Northern) blot analysis with hybridization probes from the region of the herpes simplex virus type 1 DNA known to encode the gC gene indicated that gC mRNA was produced in MP-infected HeLa cells at levels relative to other mRNAs comparable with that seen in KOS-infected cells. Comparative nucleotide sequence analysis of the gC gene from the MP and KOS strains, coupled with the results of recently reported marker rescue experiments, indicates that the inability of MP to produce gC is due to a frameshift mutation in the gC-coding sequence. Because two different (out-of-phase) open reading frames overlap the gC-coding sequence in the region of the mutation, MP mRNA can encode two gC-related polypeptides. Two polypeptides of the predicted size and precipitable by anti-gC antibodies were produced by in vitro translation of MP mRNA. These polypeptides have not been detected in extracts from infected cells with the same antibodies. Comparative nucleotide sequence analyses led to several corrections in the published sequence for the gC gene and the 17,800-molecular-weight polypeptide gene just to the right in KOS DNA. These relatively minor effects on the predicted amino code sequence of gC are tabulated.
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- 1984
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29. Direct demonstration that the abundant 6-kilobase herpes simplex virus type 1 mRNA mapping between 0.23 and 0.27 map units encodes the major capsid protein VP5
- Author
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Costa, R H, Cohen, G, Eisenberg, R, Long, D, and Wagner, E
- Abstract
The two partially colinear 6-kilobase (kb) and 1.5-kb mRNAs mapping between 0.23 and 0.27 map units on the herpes simplex virus type 1 genome were precisely located. The 5' end of the 6-kb mRNA was located 28 bases downstream of the sequence ATATATT and was 10 bases to the left of the BamHI site at 0.268. This position is ca. 90 bases to the left of our earlier reported sequence (R. J. Frink, K. G. Draper, and E. K. Wagner, Proc. Natl. Acad. Sci. U.S.A. 78:6139-6143, 1981). We used a polyclonal antibody made against purified herpes simplex virus type 1 VP5 to demonstrate that the 155,000-dalton translation product of the 6-kb mRNA is this capsid protein. The antibody did not react with the 35,000-dalton translation product of the 1.5-kb mRNA. We also confirmed our identification of VP5 as the translation product of the 6-kb mRNA by comparison of tryptic peptides of the in vitro-translated protein and authentic VP5.
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- 1984
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30. High-resolution characterization of herpes simplex virus type 1 transcripts encoding alkaline exonuclease and a 50,000-dalton protein tentatively identified as a capsid protein
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Costa, R H, Draper, K G, Banks, L, Powell, K L, Cohen, G, Eisenberg, R, and Wagner, E K
- Abstract
Four partially overlapping mRNAs (1.9, 2.3, 3.9, and 4.5 kilobases [kb]) were located between 0.16 and 0.19 map units on the herpes simplex virus type 1 genome. Their direction of transcription was found to be from right to left. The 2.3-kb mRNA was found to be early (beta), whereas the others were late (beta gamma). Partial sequence analysis of the DNA encoding these genes indicated that the promoter for the 2.3-kb mRNA shares structural features with other early (beta) promoters. In vitro translation of hybrid-selected mRNA indicated that among the proteins these mRNAs encode are an 82,000-dalton (d) polypeptide reactive with a monoclonal antibody against herpes simplex virus type 2 alkaline exonuclease and a 50,000-d polypeptide weakly reactive with a polyclonal antibody made against the capsid protein VP19C. Further experiments suggested that the 2.3-kb mRNA encodes the 82,000-d polypeptide, whereas one (or both) of the larger mRNAs encodes the 50,000-d protein. A novel finding was that the 1.9-kb mRNA appears to share part of the translational reading frame for alkaline exonuclease, but any polypeptide it encodes does not react with the monoclonal antibody to this enzyme.
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- 1983
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31. Herpes simplex virus mRNA species mapping in EcoRI fragment I
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Hall, L M, Draper, K G, Frink, R J, Costa, R H, and Wagner, E K
- Abstract
We described the detailed characterization and high-resolution mapping of nine herpes simplex virus type 1 mRNAs encoded in EcoRI fragment I. Four of these mRNAs are partially colinear and encode the same sized polypeptide in vitro. Nucleotide sequence analysis of the DNA around the 5' ends of these mRNAs suggested that the larger may encode a small (ca. 100-dalton) polypeptide not resolvable by in vitro translation.
- Published
- 1982
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32. A liver-specific DNA-binding protein recognizes multiple nucleotide sites in regulatory regions of transthyretin, alpha 1-antitrypsin, albumin, and simian virus 40 genes.
- Author
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Costa, R H, Grayson, D R, Xanthopoulos, K G, and Darnell, J E
- Abstract
Double-stranded oligodeoxynucleotides that represent protein binding sites in the regulatory regions of the mouse genes encoding transthyretin (TTR) and alpha 1-antitrypsin (alpha 1-AT) bound a nuclear protein factor(s) found mainly in hepatocytes. A site in the regulatory region of the gene encoding rat serum albumin and, surprisingly, a region in the simian virus 40 enhancer also bind the same factor. Oligodeoxynucleotide affinity chromatography (with one of the TTR binding sites) allowed a 500-fold purification of the protein. The purified protein protected similar portions of all the regulatory regions, as well as the simian virus 40 core C enhancer element, from digestion with DNase I. A DNA-binding protein previously purified from liver by virtue of its ability to bind to several virus enhancer sequences also binds to TTR, alpha 1-AT, and albumin regulatory sites. Thus, all these binding sites, which contain only minimal sequence similarity, may bind to a single protein, or a similar family of proteins, that activates liver-specific transcription of coordinately expressed genes.
- Published
- 1988
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33. The cell-specific enhancer of the mouse transthyretin (prealbumin) gene binds a common factor at one site and a liver-specific factor(s) at two other sites
- Author
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Costa, R H, Lai, E, Grayson, D R, and Darnell, J E
- Abstract
We previously defined two distinct cell-specific DNA elements controlling the transient expression of the transthyretin gene in Hep G2 (human hepatoma) cells: a proximal promoter region (-202 base pairs [bp] to the cap site), and a far-upstream cell-specific enhancer located between 1.6 and 2.15 kilobases (kb) 5' of the cap site (R. H. Costa, E. Lai, and J. E. Darnell, Jr., Mol. Cell. Biol. 6:4697-4708, 1986). In this report, we located the effective transthyretin enhancer element within a 100-bp region between 1.96 and 1.86 kb 5' to the mRNA cap site. In Hep G2 nuclear extracts, three protein-binding sites within this minimal enhancer element were identified by gel mobility and methylation protection experiments. Each binding site was required for full enhancer activity in Hep G2 transient expression assays. Competition experiments in protein-binding assays suggested that two of the three sites were recognized by a similar factor and that the protein interaction with the third site was different. The nuclear protein(s) which bound to the two homologous sites was found mainly or only in cells of hepatic origin, suggesting an involvement of this region in the cell-specific function of this enhancer. The nuclear protein(s) recognizing the third enhancer region was also found in HeLa and spleen cells.
- Published
- 1988
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34. Characterization of the genes encoding herpes simplex virus type 1 and type 2 alkaline exonucleases and overlapping proteins
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Draper, K G, Devi-Rao, G, Costa, R H, Blair, E D, Thompson, R L, and Wagner, E K
- Abstract
A detailed sequence analysis of the herpes simplex virus type 1 (HSV-1) and HSV-2 DNA encoding the alkaline exonuclease mRNA clusters has been completed. Three partially colinear mRNAs (2.3, 1.9, and 0.9 kilobases) are completely encoded within the DNA sequence presented. The putative promoter regions of the transcripts were inserted upstream of a plasmid-borne chloramphenicol acetyl transferase (CAT) gene and assayed for their ability to induce transcription of the CAT gene upon low multiplicity of infection with HSV in transient expression assays. We conclude that the expression of all three transcripts appear to be controlled by individual promoters. The 2.3-kilobase mRNA contains an open translational reading frame sufficient to encode 626 amino acids for the HSV-1 alkaline exonuclease enzyme; this value is 620 amino acids for HSV-2. A comparison of the predicted amino acid sequences of the HSV-1 and HSV-2 alkaline exonuclease enzymes revealed significant amino acid differences in the N-terminal portions of the two proteins; however, computer analyses suggest that the three-dimensional structures of the HSV-1 and HSV-2 nuclease enzymes are very similar. The 0.9-kilobase mRNA contains an open reading frame which shares a small amount of out-of-phase overlap with the C-terminal portion of the alkaline nuclease open reading frame. This open reading frame has the capacity to encode a 96-amino-acid polypeptide (10,500 daltons).
- Published
- 1986
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35. The transcriptional activator hepatocyte nuclear factor 6 regulates liver gene expression
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Samadani, U and Costa, R H
- Abstract
The hepatocyte nuclear factor 3(alpha) (HNF-3(alpha)), -3(beta), and -3(gamma) proteins share homology in the winged-helix/fork head DNA binding domain and mediate hepatocyte-enriched transcription of numerous genes whose expression is necessary for organ function. In this work, we identify a liver-enriched transcription factor, HNF-6, which recognizes the -138 to -126 region of the HNF-3(beta) promoter and binds the original HNF-3 site of the transthyretin promoter (-94 to -106). We show that HNF-6 and HNF-3 possess different DNA binding specificities by competition and methylation interference studies and are immunologically distinct. Site-directed mutagenesis of the HNF-6 sites in the HNF-3(beta) and transthyretin promoters diminishes reporter gene expression, suggesting that HNF-6 activates transcription of these promoters. Using the HNF-6 binding sequence DHWATTGAYTWWD (where W = A or T, Y = T or C, H is not G, and D is not C) determined by sequence comparison and methylation interference, we predicted that HNF-6 will bind to 22 additional hepatocyte-enriched genes. Of these potential target genes, we selected seven of the HNF-6 binding sequences and demonstrated that they bind the HNF-6 protein. These include promoter sequences from alpha-2 urinary globulin, alpha-1 antitrypsin, cytochrome P-450 2C13, L-type 6-phosphofructo-2-kinase, mouse major urinary protein, tryptophan oxygenase, and alpha-fetoprotein genes. HNF-6 binding activity was also found in the intestinal epithelial cell line HT29, and potential HNF-6 binding sites were present in intestinal sucrase isomaltase, cdx-2 homeodomain protein, and intestinal fatty acid binding protein promoter regions. These studies suggest that HNF-6 may regulate hepatocyte-specific genes and may play a role in epithelial cell differentiation of gut endoderm via regulation of HNF-3(beta).
- Published
- 1996
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36. An unusual spliced herpes simplex virus type 1 transcript with sequence homology to Epstein-Barr virus DNA
- Author
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Costa, R H, Draper, K G, Kelly, T J, and Wagner, E K
- Abstract
High-resolution transcription mapping localized a spliced 2.7-kilobase herpes simplex virus type 1 mRNA. The 4-kilobase intron of this transcript encodes a nested set of transcripts on the opposite DNA strand. The nucleotide sequence of the DNA encoding the left-hand and right-hand exons of the spliced transcript was determined, and the salient features are presented here. Of major interest is that both exons contained regions within several hundred bases of the splice donor and acceptor sites which showed homology to two regions of the Epstein-Barr virus genome, which are themselves 3 kilobases apart. The spliced herpes simplex virus transcript encoded a translational reading frame which could encode a protein with an approximate size of 75,000 daltons. This value is in agreement with in vitro translation data. The predicted amino acid sequence of the herpes simplex virus protein had significant homology with putative amino acid sequences encoded by the homologous Epstein-Barr virus DNA sequences.
- Published
- 1985
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37. Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes
- Author
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Costa, R H, Grayson, D R, and Darnell, J E
- Abstract
Transthyretin (TTR) and alpha 1-antitrypsin (alpha 1-AT) are expressed at high levels in the liver and also in at least one other cell type. We report here a detailed analysis of the proximal regulatory region of the TTR gene, which has uncovered two new DNA-binding factors that are present mainly (or only) in hepatocytes. One of these new factors, hepatocyte nuclear factor 3 (HNF-3), binds to two sites that are crucial in TTR expression as well as to two additional sites in the alpha 1-AT proximal enhancer region. The second new factor, HNF-4, binds to two sites in TTR that are required for gene activity. We had previously identified binding sites for another hepatocyte-enriched DNA-binding protein (C/EBP or a relative thereof), and additional promoter-proximal sites for that protein in both TTR and alpha 1-AT are also reported here. From these results it seems clear that cell-specific expression is not simply the result of a single cell-specific factor for each gene but the result of a combination of such factors. The variation and distribution of such factors among different cell types could be an important basis for the coordinate expression of the TTR and alpha 1-AT genes in the liver or the discordant transcriptional activation of these genes in a few other cell types. The identification of such cell-enriched factors is a necessary prelude to understanding the basis for cell specificity.
- Published
- 1989
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38. Anaerobic side-stream reactor for excess sludge reduction: 5-year management of a full-scale plant.
- Author
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Velho VF, Foladori P, Andreottola G, and Costa RH
- Subjects
- Aerobiosis, Anaerobiosis, Bioreactors, Sewage, Waste Management methods, Wastewater
- Abstract
The long-term performances of a full-scale anaerobic side-stream reactor (ASSR) aimed at sludge reduction have been monitored for the first time, in comparison with a conventional activated sludge process (CAS). The plant was integrated with an ASSR treatment of 2293-3293 m(3). Operational parameters in the ASSR were: ORP -250 mV, interchange ratio of 7-10%, hydraulic retention time of 7 d. No worsening of effluent quality was observed in the ASSR configuration and removal efficiency of COD and NH4 was above 95%. A slight increase in the Sludge Volume Index did not cause worsening in effluent solids concentration. The observed sludge yield (Yobs) passed from 0.44 kgTSS/kgCOD in the CAS to 0.35 in the ASSR configuration. The reduction of Yobs by 20% is lower than expected from the literature where sythetic wastewater is used, indicating that sludge reduction efficiency is largely affected by inert mass fed with influent real wastewater. An increase by 45% of the ASSR volume did not promote a further reduction of Yobs, because sludge reduction is affected not solely by endogenous decay but also by other factors such as interchange ratio and aerobiosis/anaerobiosis alternation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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39. Human peroxisome proliferator-activated receptor alpha (PPARalpha) supports the induction of peroxisome proliferation in PPARalpha-deficient mouse liver.
- Author
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Yu S, Cao WQ, Kashireddy P, Meyer K, Jia Y, Hughes DE, Tan Y, Feng J, Yeldandi AV, Rao MS, Costa RH, Gonzalez FJ, and Reddy JK
- Subjects
- Animals, Cell Division drug effects, Gene Expression Regulation, Humans, Mice, Mice, Inbred C57BL, Mice, Knockout, Oxidation-Reduction, Peroxisomes drug effects, Pyrimidines pharmacology, RNA, Messenger analysis, Liver metabolism, Peroxisomes metabolism, Receptors, Cytoplasmic and Nuclear physiology, Transcription Factors physiology
- Abstract
Peroxisome proliferators, which function as peroxisome proliferator-activated receptor alpha (PPARalpha) agonists, induce peroxisomal, microsomal, and mitochondrial fatty acid oxidation enzymes, in conjunction with peroxisome proliferation, in liver cells. Sustained activation of PPARalpha leads to the development of liver tumors in rats and mice. The assertion that synthetic PPARalpha ligands pose negligible carcinogenic risk to humans is attributable, in part, to the failure to observe peroxisome proliferation in human hepatocytes. To explore the mechanism(s) of species-specific differences in response to PPARalpha ligands, we determined the functional competency of human PPARalpha in vivo and compared its potency with that of mouse PPARalpha. Recombinant adenovirus that expresses human or mouse PPARalpha was produced and administered intravenously to PPARalpha-deficient mice. Human as well as mouse PPARalpha fully restored the development of peroxisome proliferator-induced immediate pleiotropic responses, including peroxisome proliferation and enhanced expression of genes involved in lipid metabolism as well as nonperoxisomal genes, such as CD36, Ly-6D, Rbp7, monoglyceride lipase, pyruvate dehydrogenase kinase-4, and C3f, that have been identified recently to be up-regulated in livers with peroxisome proliferation. These studies establish that human PPARalpha is functionally competent and is equally as dose-sensitive as mouse PPARalpha in inducing peroxisome proliferation within the context of mouse liver environment and that it can heterodimerize with mouse retinoid X receptor, and this human PPARalpha-mouse retinoid X receptor chimeric heterodimer transcriptionally activates mouse PPARalpha target genes in a manner qualitatively similar to that of mouse PPARalpha.
- Published
- 2001
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40. DDB2 induces nuclear accumulation of the hepatitis B virus X protein independently of binding to DDB1.
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Nag A, Datta A, Yoo K, Bhattacharyya D, Chakrabortty A, Wang X, Slagle BL, Costa RH, and Raychaudhuri P
- Subjects
- G1 Phase, Hepatocytes metabolism, Humans, Protein Subunits, Tumor Cells, Cultured, Viral Regulatory and Accessory Proteins, Cell Nucleus metabolism, DNA-Binding Proteins metabolism, DNA-Binding Proteins physiology, Trans-Activators metabolism
- Abstract
The hepatitis B virus (HBV) X protein (HBx) is critical for the life cycle of the virus. HBx associates with several host cell proteins including the DDB1 subunit of the damaged-DNA binding protein DDB. Recent studies on the X protein encoded by the woodchuck hepadnavirus have provided correlative evidence indicating that the interaction with DDB1 is important for establishment of infection by the virus. In addition, the interaction with DDB1 has been implicated in the nuclear localization of HBx. Because the DDB2 subunit of DDB is required for the nuclear accumulation of DDB1, we investigated the role of DDB2 in the nuclear accumulation of HBx. Here we show that expression of DDB2 increases the nuclear levels of HBx. Several C-terminal deletion mutants of DDB2 that fail to bind DDB1 are able to associate with HBx, suggesting that DDB2 may associate with HBx independently of binding to DDB1. We also show that DDB2 enhances the nuclear accumulation of HBx independently of binding to DDB1, since a mutant that does not bind DDB1 is able to enhance the nuclear accumulation of HBx. HBV infection is associated with liver pathogenesis. We show that the nuclear levels of DDB1 and DDB2 are tightly regulated in hepatocytes. Studies with regenerating mouse liver indicate that during late G1 phase the nuclear levels of both subunits of DDB are transiently increased, followed by a sharp decrease in S phase. Taken together, these results suggest that DDB1 and DDB2 would participate in the nuclear functions of HBx effectively only during the late-G1 phase of the cell cycle.
- Published
- 2001
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41. Increased levels of forkhead box M1B transcription factor in transgenic mouse hepatocytes prevent age-related proliferation defects in regenerating liver.
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Wang X, Quail E, Hung NJ, Tan Y, Ye H, and Costa RH
- Subjects
- Aging genetics, Animals, Cell Division, Cyclin-Dependent Kinases metabolism, Cyclophilins genetics, DNA Replication, Forkhead Box Protein M1, Forkhead Transcription Factors, Hepatocytes physiology, Mice, Mice, Transgenic, Phosphorylation, RNA Probes, RNA, Antisense, Retinoblastoma Protein metabolism, Ribonucleases, S Phase, Transfection, Aging physiology, DNA-Binding Proteins genetics, Hepatocytes cytology, Liver physiology, Liver Regeneration physiology, Transcription Factors genetics
- Abstract
The forkhead box (Fox) family of transcription factors share homology in the winged helix/forkhead DNA-binding domain and play important roles in regulating cellular proliferation, differentiation, longevity, and cellular transformation. Forkhead box M1B (FoxM1B) is a ubiquitously expressed member of the Fox transcription factor family whose expression is restricted to proliferating cells and that mediates hepatocyte entry into DNA synthesis and mitosis during liver regeneration. Recent cDNA microarray studies indicated that age-related defects in cellular proliferation are associated with diminished expression of the FoxM1B transcription factor. Here, we show that increased levels of FoxM1B in regenerating liver of old transgenic mice restore the sharp peaks in hepatocyte DNA replication and mitosis that are the hallmarks of young regenerating mouse liver. Restoration of the young regenerating liver phenotype is associated with increased expression of numerous cell cycle regulatory genes that include cyclin D1, cyclin A2, cyclin F, cyclin B1, cyclin B2, Cdc25B, and p55cdc. Cotransfection assays in the human hepatoma HepG2 cell line demonstrated that FoxM1B protein stimulated expression of both the cyclin B1 and cyclin D1 promoters, suggesting that these cyclin genes are a direct FoxM1B transcriptional target. These results suggest that FoxM1B controls the transcriptional network of genes that are essential for cell division and exit from mitosis. Our results indicate that reduced expression of the FoxM1B transcription factor contributes to the decline in cellular proliferation observed in the aging process.
- Published
- 2001
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42. Defects in pulmonary vasculature and perinatal lung hemorrhage in mice heterozygous null for the Forkhead Box f1 transcription factor.
- Author
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Kalinichenko VV, Lim L, Stolz DB, Shin B, Rausa FM, Clark J, Whitsett JA, Watkins SC, and Costa RH
- Subjects
- Alleles, Animals, Apoptosis, Blotting, Western, Bone Morphogenetic Protein 4, Bone Morphogenetic Proteins metabolism, DNA, Complementary metabolism, Dose-Response Relationship, Drug, Endothelial Growth Factors metabolism, Endothelium metabolism, Forkhead Transcription Factors, Hemorrhage, Heterozygote, Immunohistochemistry, In Situ Nick-End Labeling, Lung pathology, Lymphokines metabolism, Mice, Mice, Knockout, Microscopy, Electron, Models, Genetic, Mutation, Oligonucleotide Array Sequence Analysis, Phenotype, Platelet Endothelial Cell Adhesion Molecule-1 metabolism, RNA, Messenger metabolism, Receptor Protein-Tyrosine Kinases metabolism, Receptors, Growth Factor metabolism, Receptors, Vascular Endothelial Growth Factor, Trans-Activators metabolism, Transcription Factors physiology, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, beta-Galactosidase metabolism, DNA-Binding Proteins, Lung embryology, Lung metabolism, Transcription Factors biosynthesis, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Decreased pulmonary expression of Forkhead Box f1 (Foxf1) transcription factor was associated with lethal alveolar hemorrhage in 55% of the Foxf1 +/- newborn mice. The severity of the pulmonary abnormalities correlates with the levels of Foxf1 mRNA. Defects in alveolarization and vasculogenesis were observed in subsets of the Foxf1 +/- mice with relatively low levels of expression from the normal Foxf1 allele. Lung hemorrhage was coincident with disruption of the mesenchymal-epithelial cell interfaces in the alveolar and bronchiolar regions of the lung parenchyma and was associated with increased apoptosis and reduced surfactant protein B (SP-B) expression. Finally, the lung defect associated with the Foxf1 +/- mutation was accompanied by reduced expression of vascular endothelial growth factor (VEGF), the VEGF receptor 2 (Flk-1), bone morphogenetic protein 4 (Bmp-4), and the transcription factors of the Brachyury T-Box family (Tbx2-Tbx5) and Lung Kruppel-like Factor. Reduction in the level of Foxf1 caused neonatal pulmonary hemorrhage and abnormalities in alveologenesis, implicating this transcription factor in the regulation of mesenchyme-epithelial interaction critical for lung morphogenesis., (Copyright 2001 Academic Press.)
- Published
- 2001
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43. Transcription factors in mouse lung development and function.
- Author
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Costa RH, Kalinichenko VV, and Lim L
- Subjects
- Amino Acid Motifs, Animals, Cell Differentiation, Epithelial Cells cytology, Epithelial Cells metabolism, Gene Expression Regulation, Developmental, Lung cytology, Mesoderm cytology, Mesoderm metabolism, Mice, Mice, Knockout, Mice, Transgenic, Morphogenesis physiology, Phenotype, Transcription Factors genetics, Lung embryology, Lung metabolism, Transcription Factors metabolism
- Abstract
Development of the mouse lung initiates on day 9.5 postcoitum from the laryngotracheal groove and involves mesenchymal-epithelial interactions, in particular, those between the splanchnic mesoderm and epithelial cells (derived from foregut endoderm) that induce cellular proliferation, migration, and differentiation, resulting in branching morphogenesis. This developmental process mediates formation of the pulmonary bronchiole tree and integrates a terminal alveolar region with an extensive endothelial capillary bed, which facilitates efficient gas exchange with the circulatory system. The major function of the mesenchymal-epithelial signaling is to potentiate the activity or expression of cell type-specific transcription factors in the developing lung, which, in turn, cooperatively bind to distinct promoter regions and activate target gene expression. In this review, we focus on the role of transcription factors in lung morphogenesis and the maintenance of differentiated gene expression. These lung transcription factors include forkhead box A2 [also known as hepatocyte nuclear factor (HNF)-3beta], HNF-3/forkhead homolog (HFH)-8 [also known as FoxF1 or forkhead-related activator-1], HNF-3/forkhead homolog-4 (also known as FoxJ1), thyroid transcription factor-1 (Nkx2.1), and homeodomain box A5 transcription factors, the zinc finger Gli (mouse homologs of the Drosophila cubitus interruptus) and GATA transcription factors, and the basic helix-loop-helix Pod1 transcription factor. We summarize the phenotypes of transgenic and knockout mouse models, which define important functions of these transcription factors in cellular differentiation and lung branching morphogenesis.
- Published
- 2001
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44. Differential expression of forkhead box transcription factors following butylated hydroxytoluene lung injury.
- Author
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Kalinichenko VV, Lim L, Shin B, and Costa RH
- Subjects
- Animals, Bronchi metabolism, Bronchi pathology, Butylated Hydroxytoluene, Cell Division physiology, Endothelium metabolism, Endothelium pathology, Forkhead Box Protein M1, Forkhead Transcription Factors, Hepatocyte Nuclear Factor 3-beta, Lung metabolism, Lung pathology, Lung Diseases chemically induced, Lung Diseases pathology, Male, Mesoderm pathology, Mice, Mice, Inbred BALB C, Muscle, Smooth cytology, Muscle, Smooth metabolism, DNA-Binding Proteins metabolism, Lung Diseases metabolism, Nuclear Proteins metabolism, Trans-Activators metabolism, Transcription Factors metabolism
- Abstract
The forkhead box (Fox) proteins are a growing family of transcription factors that have important roles in cellular proliferation and differentiation and in organ morphogenesis. The Fox family members hepatocyte nuclear factor (HNF)-3beta (Foxa2) and HNF-3/forkhead homolog (HFH)-8 (FREAC-1, Foxf1) are expressed in adult pulmonary epithelial and mesenchymal cells, respectively, but these cells display only low expression levels of the proliferation-specific HFH-11B gene (Trident, Foxm1b). The regulation of these Fox transcription factors in response to acute lung injury, however, has yet to be determined. We report here on the use of butylated hydroxytoluene (BHT)-mediated lung injury to demonstrate that HFH-11 protein and RNA levels were markedly increased throughout the period of lung repair. The maximum levels of HFH-11 were observed by day 2 following BHT injury when both bronchiolar and alveolar epithelial cells were undergoing extensive proliferation. Although BHT lung injury did not alter epithelial cell expression of HNF-3beta, a 65% reduction in HFH-8 mRNA levels was observed during the period of mesenchymal cell proliferation. HFH-8-expressing cells were colocalized with platelet endothelial cell adhesion molecule-1-positive alveolar endothelial cells and with alpha-smooth muscle actin-positive peribronchiolar smooth muscle cells.
- Published
- 2001
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45. Elevated levels of hepatocyte nuclear factor 3beta in mouse hepatocytes influence expression of genes involved in bile acid and glucose homeostasis.
- Author
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Rausa FM, Tan Y, Zhou H, Yoo KW, Stolz DB, Watkins SC, Franks RR, Unterman TG, and Costa RH
- Subjects
- ATP Binding Cassette Transporter, Subfamily B metabolism, ATP-Binding Cassette Transporters metabolism, Animals, Base Sequence, Bile Acids and Salts metabolism, Blotting, Western, Carrier Proteins metabolism, Cell Line, DNA Methylation, Glucose metabolism, Glutathione Transferase metabolism, Glycogen metabolism, Hepatocyte Nuclear Factor 3-beta, Hepatocyte Nuclear Factor 6, Homeodomain Proteins metabolism, Immunohistochemistry, Insulin-Like Growth Factor Binding Protein 1 blood, Insulin-Like Growth Factor Binding Protein 1 metabolism, Ligands, Liver embryology, Liver metabolism, Liver pathology, Mice, Mice, Transgenic, Microscopy, Electron, Models, Genetic, Molecular Sequence Data, Organic Anion Transporters, Sodium-Dependent, Phenotype, Prealbumin genetics, Prealbumin metabolism, Promoter Regions, Genetic, Protein Isoforms, Recombinant Proteins metabolism, Symporters, Time Factors, Trans-Activators metabolism, Transcription, Genetic, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Liver cytology, Membrane Transport Proteins, Nuclear Proteins genetics, Nuclear Proteins metabolism, Transcription Factors
- Abstract
The winged helix transcription factor, hepatocyte nuclear factor-3beta (HNF-3beta), mediates the hepatocyte-specific transcription of numerous genes important for liver function. However, the in vivo role of HNF-3beta in regulating these genes remains unknown because homozygous null HNF3beta mouse embryos die in utero prior to liver formation. In order to examine the regulatory function of HNF-3beta, we created transgenic mice in which the -3-kb transthyretin promoter functions to increase hepatocyte expression of the rat HNF-3beta protein. Postnatal transgenic mice exhibit growth retardation, depletion of hepatocyte glycogen storage, and elevated levels of bile acids in serum. The retarded growth phenotype is likely due to a 20-fold increase in hepatic expression of insulin-like growth factor binding protein 1 (IGFBP-1), which results in elevated levels in serum of IGFBP-1 and limits the biological availability of IGFs required for postnatal growth. The defects in glycogen storage and serum bile acids coincide with diminished postnatal expression of hepatocyte genes involved in gluconeogenesis (phosphoenolpyruvate carboxykinase and glycogen synthase) and sinusoidal bile acid uptake (Ntcp), respectively. These changes in gene transcription may result from the disruptive effect of HNF-3beta on the hepatic expression of the endogenous mouse HNF-3alpha,-3beta, -3gamma, and -6 transcription factors. Furthermore, adult transgenic livers lack expression of the canalicular phospholipid transporter, mdr2, which is consistent with ultrastructure evidence of damage to transgenic hepatocytes and bile canaliculi. These transgenic studies represent the first in vivo demonstration that the HNF-3beta transcriptional network regulates expression of hepatocyte-specific genes required for bile acid and glucose homeostasis, as well as postnatal growth.
- Published
- 2000
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46. The nuclear receptor fetoprotein transcription factor is coexpressed with its target gene HNF-3beta in the developing murine liver, intestine and pancreas.
- Author
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Rausa FM, Galarneau L, Bélanger L, and Costa RH
- Subjects
- Animals, Carcinoma, Hepatocellular genetics, DNA-Binding Proteins metabolism, Endoderm, Gene Expression Regulation, Developmental, Hepatocyte Nuclear Factor 3-beta, Intestines growth & development, Liver growth & development, Mice, Nuclear Proteins metabolism, Pancreas growth & development, Receptors, Cytoplasmic and Nuclear genetics, Receptors, Cytoplasmic and Nuclear metabolism, Transcription Factors metabolism, Tumor Cells, Cultured, DNA-Binding Proteins genetics, Intestines embryology, Liver embryology, Nuclear Proteins genetics, Pancreas embryology, Transcription Factors genetics
- Abstract
During organogenesis, the winged helix hepatocyte nuclear factor 3beta (HNF-3beta) protein participates in regulating gene transcription in the developing esophagus, trachea, liver, lung, pancreas, and intestine. Hepatoma cell transfection studies identified a critical HNF-3beta promoter factor, named UF2-H3beta, and here, we demonstrate that UF2-H3beta is identical to the fetoprotein transcription factor (FTF). In situ hybridization studies of mouse embryos demonstrate that FTF expression initiates in the foregut endoderm during liver and pancreatic morphogenesis (day 9) and that earlier expression of FTF is observed in the yolk sac endoderm, branchial arch and neural crest cells (day 8). Abundant FTF hybridization signals are observed throughout morphogenesis of the liver, pancreas, and intestine and its expression continues in the epithelial cells of these adult organs. In day 17 mouse embryos and adult pancreas, however, expression of FTF becomes restricted to the exocrine acinar and ductal epithelial cells.
- Published
- 1999
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47. Premature expression of the winged helix transcription factor HFH-11B in regenerating mouse liver accelerates hepatocyte entry into S phase.
- Author
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Ye H, Holterman AX, Yoo KW, Franks RR, and Costa RH
- Subjects
- Animals, CCAAT-Enhancer-Binding Proteins, Cell Nucleus metabolism, Cyclins biosynthesis, Cyclins genetics, DNA Replication, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins genetics, Forkhead Box Protein M1, Forkhead Transcription Factors, Humans, Liver cytology, Male, Mice, Mice, Transgenic, Mitosis, Nuclear Proteins biosynthesis, Phosphoproteins biosynthesis, Phosphoproteins genetics, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, S Phase, Signal Transduction, Time Factors, Transcription Factors genetics, X-ray Repair Cross Complementing Protein 1, Liver metabolism, Liver Regeneration physiology, Transcription Factors biosynthesis
- Abstract
Two-thirds partial hepatectomy (PH) induces differentiated cells in the liver remnant to proliferate and regenerate to its original size. The proliferation-specific HNF-3/fork head homolog-11B protein (HFH-11B; also known as Trident and Win) is a family member of the winged helix/fork head transcription factors and in regenerating liver its expression is reactivated prior to hepatocyte entry into DNA replication (S phase). To examine whether HFH-11B regulates hepatocyte proliferation during liver regeneration, we used the -3-kb transthyretin (TTR) promoter to create transgenic mice that displayed ectopic hepatocyte expression of HFH-11B. Liver regeneration studies with the TTR-HFH-11B mice demonstrate that its premature expression resulted in an 8-h acceleration in the onset of hepatocyte DNA replication and mitosis. This liver regeneration phenotype is associated with protracted expression of cyclin D1 and C/EBPbeta, which are involved in stimulating DNA replication and premature expression of M phase promoting cyclin B1 and cdc2. Consistent with the early hepatocyte entry into S phase, regenerating transgenic livers exhibited earlier expression of DNA repair genes (XRCC1, mHR21spA, and mHR23B). Furthermore, in nonregenerating transgenic livers, ectopic HFH-11B expression did not elicit abnormal hepatocyte proliferation, a finding consistent with the retention of the HFH-11B transgene protein in the cytoplasm. We found that nuclear translocation of the HFH-11B transgene protein requires mitogenic signalling induced by PH and that its premature availability in regenerating transgenic liver allowed nuclear translocation to occur 8 h earlier than in wild type.
- Published
- 1999
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48. HNF-3/forkhead homologue-4 influences lung morphogenesis and respiratory epithelial cell differentiation in vivo.
- Author
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Tichelaar JW, Lim L, Costa RH, and Whitsett JA
- Subjects
- Animals, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors, Cell Differentiation physiology, Cells, Cultured, Epithelial Cells cytology, Epithelial Cells physiology, Gene Expression Regulation, Developmental physiology, Hepatocyte Nuclear Factor 4, Lung cytology, Mice, Mice, Transgenic, Morphogenesis physiology, Respiratory System cytology, DNA-Binding Proteins, Lung embryology, Phosphoproteins physiology, Transcription Factors physiology
- Abstract
HNF-3/forkhead homologue 4 (HFH-4), a transcription factor of the winged helix/forkhead family, is expressed in various tissues including lung, brain, oviduct, testis, and embryonic kidney. In order to test whether the temporospatial expression of HFH-4 influences lung morphogenesis, HFH-4 was expressed in lungs of transgenic mice under control of the surfactant protein C (SP-C) promoter. The morphology of the lungs from SP-C/HFH-4 embryos (day 18 postconception) was distinctly abnormal, and the severity of the alterations correlated with the level of transgene expression as detected by in situ hybridization. At high levels of expression, HFH-4 altered epithelial cell differentiation and inhibited branching morphogenesis. Atypical cuboidal or columnar cells lined the lung periphery of SP-C/HFH-4 transgenic mice. The atypical epithelial cells seen in the SP-C/HFH-4 mice expressed thyroid transcription factor-1 and hepatocyte nuclear factor 3beta (HNF-3beta). However, surfactant proteins SP-B, SP-C, and Clara cell secretory protein, normally produced by nonciliated epithelial cells in lung parenchyma were lacking. beta-Tubulin IV, a marker of ciliated cells, stained the atypical columnar cells produced by expression of high levels of the SP-C/HFH-4 transgene. Ectopic expression of HFH-4 in developing mouse lung altered epithelial cell differentiation and morphology, restricting the expression of markers typical of nonciliated cells of the distal lung parenchyma., (Copyright 1999 Academic Press.)
- Published
- 1999
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49. HNF-3/forkhead homologue-4 (HFH-4) is expressed in ciliated epithelial cells in the developing mouse lung.
- Author
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Tichelaar JW, Wert SE, Costa RH, Kimura S, and Whitsett JA
- Subjects
- Animals, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins metabolism, Epithelial Cells metabolism, Hepatocyte Nuclear Factor 4, Immunohistochemistry, Mice, Mice, Knockout, Nuclear Proteins biosynthesis, Nuclear Proteins genetics, Phosphoproteins metabolism, Protein Biosynthesis, Thyroid Nuclear Factor 1, Tissue Distribution, Transcription Factors genetics, Transcription Factors metabolism, Tubulin biosynthesis, Tubulin metabolism, Lung embryology, Lung metabolism, Phosphoproteins biosynthesis, Transcription Factors biosynthesis, Uteroglobin
- Abstract
HNF-3/forkhead homologue-4 (HFH-4), a transcription factor of the winged-helix/forkhead family, was detected by immunohistochemistry in tissue of the developing mouse. HFH-4 protein was present in epithelial cells of the lung, trachea, oviduct, and embryonic esophagus, and in ependymal cells lining the spinal column and ventricles of the brain. In lung, trachea, and nose, HFH-4 was expressed in a distinct subset of epithelial cells that also expressed beta-tubulin IV, a ciliated cell marker. Cellular sites of HFH-4 and beta-tubulin IV expression were distinct from that of Clara cell secretory protein (CCSP), which was detected in nonciliated epithelial cells in the conducting airway of the lung. HFH-4 and beta-tubulin IV, but not CCSP, were detected in the respiratory epithelium of thyroid transcription factor-1 (TTF-1) gene-targeted mice. The presence of HFH-4 and beta-tubulin IV in TTF-1 gene-targeted mice demonstrates that differentiation of ciliated epithelium does not require TTF-1. Co-localization of HFH-4 and beta-tubulin IV staining in various tissues during mouse development supports a role for HFH-4 in the differentiation of ciliated cell lineages.
- Published
- 1999
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50. The cut-homeodomain transcriptional activator HNF-6 is coexpressed with its target gene HNF-3 beta in the developing murine liver and pancreas.
- Author
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Rausa F, Samadani U, Ye H, Lim L, Fletcher CF, Jenkins NA, Copeland NG, and Costa RH
- Subjects
- Amino Acid Sequence, Animals, Carcinoma, Hepatocellular pathology, Cell Differentiation, Crosses, Genetic, DNA-Binding Proteins biosynthesis, Female, Fetal Proteins biosynthesis, Fetal Proteins genetics, Hepatocyte Nuclear Factor 3-beta, Hepatocyte Nuclear Factor 6, Homeodomain Proteins biosynthesis, Homeodomain Proteins genetics, Humans, In Situ Hybridization, Intestinal Mucosa embryology, Intestinal Mucosa metabolism, Islets of Langerhans embryology, Islets of Langerhans metabolism, Liver metabolism, Liver Neoplasms pathology, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Multigene Family, Muridae genetics, Nuclear Proteins biosynthesis, Organ Specificity, Pancreas metabolism, Promoter Regions, Genetic, Sequence Alignment, Sequence Homology, Amino Acid, Trans-Activators biosynthesis, Trans-Activators genetics, Transcription, Genetic, Tumor Cells, Cultured, DNA-Binding Proteins genetics, Fetal Proteins physiology, Gene Expression Regulation, Developmental, Genes, Homeobox, Homeodomain Proteins physiology, Liver embryology, Nuclear Proteins genetics, Pancreas enzymology, Trans-Activators physiology, Transcription Factors
- Abstract
Murine hepatocyte nuclear factor-3 beta (HNF-3 beta) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork head DNA binding domain and that participate in embryonic pattern formation. HNF-3 beta also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal and bronchiolar epithelial, and pancreatic acinar cells. We have previously identified a liver-enriched transcription factor, HNF-6, which is required for HNF-3 beta promoter activity and also recognizes the regulatory region of numerous hepatocyte-specific genes. In this study we used the yeast one-hybrid system to isolate the HNF-6 cDNA, which encodes a cut-homeodomain-containing transcription factor that binds with the same specificity as the liver HNF-6 protein. Cotransfection assays demonstrate that HNF-6 activates expression of a reporter gene driven by the HNF-6 binding site from either the HNF-3 beta or transthyretin (TTR) promoter regions. We used interspecific backcross analysis to determine that murine Hnf6 gene is located in the middle of mouse chromosome 9. In situ hybridization studies of staged specific embryos demonstrate that HNF-6 and its potential target gene, HNF-3 beta, are coexpressed in the pancreatic and hepatic diverticulum. More detailed analysis of HNF-6 and HNF-3 beta's developmental expression patterns provides evidence of colocalization in hepatocytes, intestinal epithelial, and in the pancreatic ductal epithelial and exocrine acinar cells. The expression patterns of these two transcription factors do not overlap in other endoderm-derived tissues or the neurotube. We also found that HNF-6 is also abundantly expressed in the dorsal root ganglia, the marginal layer, and the midbrain. At day 18 of gestation and in the adult pancreas, HNF-6 and HNF-3 beta transcripts colocalize in the exocrine acinar cells, but their expression patterns diverge in other pancreatic epithelium. HNF-6, but not HNF-3 beta, expression continues in the pancreatic ductal epithelium, whereas only HNF-3 beta becomes restricted to the endocrine cells of the islets of Langerhans. We discuss these expression patterns with respect to specification of hepatocytes and differentiation of the endocrine and exocrine pancreas.
- Published
- 1997
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