Thesis (Ph.D.)--University of Adelaide, Dept. of Surgery, 1999? / Errata is tipped in behind bibliography. Bibliography: leaves 191-215.

Thesis (Ph.D.)--University of Adelaide, Dept. of Surgery, 2003. / "June 2003" Errata slip inserted in back. Includes bibliographical references (leaves 268-300). Also available online.

Thesis (Ph.D.)--University of Adelaide, Dept. of Surgery, 2003. / Title from screen page; viewed 8 Feb 2005. "June 2003" Includes bibliographical references. Also available in print form.

Die Deckzellen des Urothels sind die einzigen Zellen, die mit dem Urin über einen längeren Zeitraum in direktem Kontakt stehen. Ihre apikale Plasmamembran weist ein facettiertes Aussehen auf und hat dicht gepackte Uroplakin III Moleküle eingelagert, die durch Ausbildung von hexagonalen Proteinkomplexen entscheidend sind für die hohe chemische Resistenz, gegen die zum Teil zellschädigende Zusammensetztung des Urins. Unterhalb der apikalen Plasmamembran befinden sich zahlreiche diskuide Vesikel. Diese entsprechen in ihrem molekularen Membranaufbau der apikalen Membran und können als präapikale Reservemembran angesehen werden, die bei Bedarf in die apikale Plasmamembran eingebaut werden kann. In der vorliegenden Arbeit ist untersucht worden, in wieweit SNARE-Proteine für die Fusion zwischen den diskuiden Vesikeln und der apikalen Plasmamembran von Bedeutung sind. In Immunoblotanalysen wurde das Vorkommen von Synaptobrevin, Syntaxin, SNAP23, NSF und alpha,beta-SNAP gezeigt und es konnte in Immunopräzipitationen die Ausbildung des SNARE-Haftkomplexes nachgewiesen werden. Immunfluoreszenzuntersuchungen und Immunogoldmarkierungen bestätigten das Vorhandensein von SNARE-Proteinen und wiesen durch das subzelluläre Verteilungsmuster der Proteine auf einen kombinierten homotypischen und heterotypischen Fusionsmechanismus hin. Hinweise auf eine mit der Fusion der Vesikel gleichgeschaltete Endozytose wurden nicht gefunden, das Abknospen von Membranbestandteilen der apikalen Zellmembran konnte jedoch ultrastrukturell gezeigt werden. Ein weiterer Hinweis auf eine möglicherweise kontinuierliche Erneuerung der apikalen Zellmembran der Deckzellen konnte durch den Nachweis von Uroplakin III im Urin von gesunden Probanden gebracht werden. In einem Zentrifugationsexperiment wurde Uroplakin III in Fraktionen von Membranaggregaten sehr unterschiedlicher Größe nachgewiesen, was die ultrastrukturellen Befunde, die das Abknospen sowohl von einzelnen Vesikeln, wie auch von größeren Membranbestandteilen zeigten, bestätigt. / The luminal surface of the bladder epitehlium is continuously exposed to the urine that differs in ionic composition and osmolarlity from blodd. The apical plasma membrane of facet cells, facing the urine, is covered with rigid-looking plaques consisting of hexagonal uroplakin-particles. Together with tight junctions these plaques form a specialized membrane compartment that represents one of the tightest and most impermeable barrier of the body. Plaques also occur in the membrane of cytoplasmic discuid vesicles. Here we show that synaptobrevin, SNAP23, syntaxin, NSF and alpha/beta-SNAP are perfectly colocalized with uroplakin III at the apical plasma membrane and with the membrane of discuid vesicles. This distribution suggest that discuid vesicles of a homotypic and heterotypic fusion events. Furthermore we detected uroplakin III containing membranes of different size in the urine of healthy humans and rats. probably facet cells maintain their permeability barrier by a process of membrane renewal where pieces of the apical membrane are continuously taken off by freshly fused discuid vesicles.

No description available.

Uroplakin 1B (UPKIB) is an integral structural protein interacting with uroplakins 1A, 2 and 3 to form hexameric plaques along the bladder lumen in the asymmetric unit membrane of urothelial umbrella cells in humans and other mammals. UPKIB mRNA expression is deregulated in transitional cell carcinomas (TCCs), however the mechanisms of regulation of UPKIB have not been established. Using genome databases, a Xenopus UPKIB homologue was identified. Maximum Parsimony and BAMBE (Bayesian Analysis in Molecular Biology and Evolution) data support a close evolutionary relationship between mammalian and amphibian UPKIB mRNA. Using Unigene, UPKIB human expressed sequence tags were identified in tissues including brain, skeletal muscle and liver, suggesting the relatively widespread distribution of this membrane protein. The UPKIB genomic structure was also deduced using genome databases. Contig AC083800, identified in a high throughput genomic sequence database, spanned UPKIB and 9 exons and 8 introns were defined. A 67bp 5' untranslated region was identified using 5' rapid amplification of cDNA ends. This product was sequenced and a putative UPKIB promoter and transcription start site was deduced. Contig AC083800 spanned the transcription start site and putative promoter. Transcription factor binding motif prediction programs detected no TATA box, but did predict a CCAAT box and several binding motifs including 4 Sp-1 sites and a NFKB site. A weak CpG island was identified within a 0.5kb region including the putative promoter, exon 1 and intron 1, which was 54% GC rich with CpG:GpC ratio of 0.46, containing 15 CpG dinucleotides. Seven TCC cell lines and five peripheral blood lymphocyte samples were analysed for UPKIB expression using RT-PCR and two cell lines expressed UPKIB transcripts. Eleven CpG sites in the putative promoter were investigated for methylation using bisulfite modification analysis in normal PBL, TCC cell lines and patient TCC samples. An inverse correlation was established in TCC cell lines between UPKIB mRNA expression and degree of methylation. 5-Aza-2'deoxycytidine induced UPKIB mRNA expression in T24 cells, previously observed not to express UPKIB. Sequence analysis of patient samples revealed more complex CpG methylation patterns, reflecting tumour heterogeneity. In summary, the uroplakin 1B gene has been characterised and one mechanism of regulation of gene expression involves methylation. / Thesis (Ph.D.)--Dept of Surgery, 2003.

Errata is tipped in behind bibliography. / Bibliography: leaves 191-215. / xiv, 216, [29] leaves, [81] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Reports the partial cloning and characterisation of the human uroplakin 1B gene which has allowed analysis and characterisation of the gene with regard to its structure, chromosomal localisation and integrity. / Thesis (Ph.D.)--University of Adelaide, Dept. of Surgery, 1999?

Uroplakin 1B (UPKIB) is an integral structural protein interacting with uroplakins 1A, 2 and 3 to form hexameric plaques along the bladder lumen in the asymmetric unit membrane of urothelial umbrella cells in humans and other mammals. UPKIB mRNA expression is deregulated in transitional cell carcinomas (TCCs), however the mechanisms of regulation of UPKIB have not been established. Using genome databases, a Xenopus UPKIB homologue was identified. Maximum Parsimony and BAMBE (Bayesian Analysis in Molecular Biology and Evolution) data support a close evolutionary relationship between mammalian and amphibian UPKIB mRNA. Using Unigene, UPKIB human expressed sequence tags were identified in tissues including brain, skeletal muscle and liver, suggesting the relatively widespread distribution of this membrane protein. The UPKIB genomic structure was also deduced using genome databases. Contig AC083800, identified in a high throughput genomic sequence database, spanned UPKIB and 9 exons and 8 introns were defined. A 67bp 5' untranslated region was identified using 5' rapid amplification of cDNA ends. This product was sequenced and a putative UPKIB promoter and transcription start site was deduced. Contig AC083800 spanned the transcription start site and putative promoter. Transcription factor binding motif prediction programs detected no TATA box, but did predict a CCAAT box and several binding motifs including 4 Sp-1 sites and a NFKB site. A weak CpG island was identified within a 0.5kb region including the putative promoter, exon 1 and intron 1, which was 54% GC rich with CpG:GpC ratio of 0.46, containing 15 CpG dinucleotides. Seven TCC cell lines and five peripheral blood lymphocyte samples were analysed for UPKIB expression using RT-PCR and two cell lines expressed UPKIB transcripts. Eleven CpG sites in the putative promoter were investigated for methylation using bisulfite modification analysis in normal PBL, TCC cell lines and patient TCC samples. An inverse correlation was established in TCC cell lines between UPKIB mRNA expression and degree of methylation. 5-Aza-2'deoxycytidine induced UPKIB mRNA expression in T24 cells, previously observed not to express UPKIB. Sequence analysis of patient samples revealed more complex CpG methylation patterns, reflecting tumour heterogeneity. In summary, the uroplakin 1B gene has been characterised and one mechanism of regulation of gene expression involves methylation. / Thesis (Ph.D.)--Dept of Surgery, 2003.

Made available in DSpace on 2016-12-23T13:53:40Z (GMT). No. of bitstreams: 1 Maria Aparecida da Silva.pdf: 3366725 bytes, checksum: 59d418ef608b4e75da9a6ba6a1fdb233 (MD5) Previous issue date: 2012-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Bovine enzootic hematuria (BEH) is caused by chronic ingestion of Pteridium aquilinum and is characterized by the presence of blood in the urine and development of lesions in the urinary bladder and is responsible for economic losses. Poisoning by this plant can also occur in humans. The objective was to evaluate the lesions in bladders of animals with BEH in the south region of the Espírito Santo. For this, were evaluated 350 bladders of bovines in a slaughterhouse and, of these, selected 46 that had macroscopic lesions and/or hematuria. Samples of each bladder were fixed in formalin 10% submitted to histological processing and classified by histomorphology. The immunohistochemistry was performed with anti-vimentin, anti-cytokeratin, anti-CD31, anti-VEGF and anti-uroplakin only in the 26 bladders that presented neoplastic lesions. Non-neoplastic lesions were observed in 100% of samples and the neoplastic in 56.52%. The presence of tumors was significant (p<0.05) in the caudal portion of the bladder. Detected neoplastic types were urothelial carcinoma, in situ carcinoma, adenocarcinoma, hemangioma, myxoma and hemangiosarcoma. There was a higher frequency of dysplasia, clear cell metaplasia, inflammation and vascular thickening in bladders with neoplasm. The expression of cytokeratin was significant (p<0.05) in epithelial neoplasms and vimentin in mesenchymal neoplasms. Uroplakin III differed in varied types of neoplastic lesions and showed to be typical and atypical while that of CD31 was significantly (p <0.05) in vascular mesenchymal neoplasms. A significant difference (p <0.05) in the number of vessels extratumorais stained by VEGF between myxomas and adenocarcinomas, and in intratumoral vessels stained by CD31 and VEGF in the different tumor types. Positive correlation existed between the number of intra- and extratumoral vessels in hemangiomas, hemangiosarcomas, and myxomas stained by CD31; between hemangiomas, myxomas, and adenocarcinomas stained with VEGF; between the expression of vimentin and CD31 and between cytokeratin and uroplakin. It is concluded that bladders from bovines with BEH have non-neoplastic and neoplastic lesions, isolated or associated. Biomarkers aid in the differentiation of the histogenesis of epithelial and vascular mesenchymal neoplasms. Uroplakin demonstrated to be effective for the assessment of integrity urothelial, and vascular markers (CD31 and VEGF) for endothelial integrity and for prognosis / A hematúria enzoótica bovina (HEB) é causada pela ingestão crônica de Pteridium aquilinum e caracteriza-se pela presença de sangue na urina e desenvolvimento de lesões na bexiga, sendo responsável por grandes perdas econômicas. A intoxicação por esta planta também pode ocorrer em humanos. Objetivou-se avaliar lesões de bexigas de animais com HEB na região sul do Espírito Santo. Para isto, foram avaliadas 350 bexigas de bovinos em matadouro frigorífico e, destas, selecionadas 46 que apresentavam lesões macroscópicas e/ou hematúria. Amostras de cada bexiga foram fixadas em formol a 10% submetidas ao processamento histológico de rotina e classificadas histomorfologicamente. A imunoistoquímica foi realizada com anti-vimentina, anti-citoqueratina, anti-CD31, anti-VEGF e anti-uroplaquina apenas nas 26 bexigas que revelaram neoplasia. Lesões não neoplásicas foram observadas em 100% das amostras e neoplásicas em 56,52%. A presença de neoplasias foi significativa (p<0,05) na porção caudal da bexiga. As neoplasias encontradas foram carcinoma urotelial; carcinoma in situ, adenocarcinoma, hemangioma, mixoma e hemangiossarcoma. Houve maior frequência de displasia, metaplasia de células claras, inflamação e espessamento vascular em bexigas com neoplasia. A expressão de citoqueratina foi significativa (p<0,05) nas neoplasias epiteliais e vimentina nas mesenquimais. A marcação da uroplaquina III diferiu nos diversos tipos neoplásicos e revelou-se típica e atípica enquanto que a do CD31 foi significativa (p<0,05) nas neoplasias mesenquimais vasculares. Houve diferença significativa (p<0,05) na quantidade de vasos extratumorais marcados pelo VEGF entre os mixomas e adenocarcinomas e nos vasos intratumorais marcados por CD31 e VEGF nos diferentes tipos tumorais. Houve correlação positiva entre os vasos extra e intratumorais nos hemangiomas, hemangiossarcomas e mixomas marcados pelo CD31; nos hemangiomas, mixomas e adenocarcinomas marcados pelo VEGF; entre a expressão de vimentina e CD31 e entre citoqueratina e uroplaquina. Conclui-se que bexigas de bovinos com HEB apresentam lesões não neoplásicas e neoplásicas, isoladas ou associadas. Os biomarcadores auxiliam na diferenciação da histogênese das neoplasias epiteliais e mesenquimais vasculares. Uroplaquina demonstrou-se efetiva para a avaliação da integridade urotelial e os marcadores vasculares (CD31 e VEGF) para a integridade endotelial e para o prognóstico

No description available.