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Análise do comprimento ureteral em cadáveres adultosNovaes, Hugo Fabiano Fernandes de January 2012 (has links)
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Previous issue date: 2012 / Em algumas situações, correlações entre diferentes estruturas do corpo humano poderiam auxiliar no planejamento de cirurgias intra-abdominais. O conhecimento do comprimento do ureter permitiria um planejamento cirúrgico pré-operatório, reduzindo gastos com exames; auxiliaria também na escolha do cateter duplo-J, reduzindo sintomatologia e morbidade, aumentando a aderência ao tratamento. Objetivo: Avaliar o comprimento ureteral em cadáveres adultos e analisar suas correlações com determinadas medidas antropométricas. Desenho do estudo: estudo transversal, descritivo e analítico. Materiais e métodos: realizamos mensuração do comprimento ureteral de cadáveres adultos, encaminhados para necropsia entre abril de 2009 e janeiro de 2012. Adicionalmente, coletamos as seguintes medidas: altura, distância ombro-punho, cotovelo-punho, xifo-umbilical, distância umbigo-púbis, distância xifo-púbica e distância entre espinhas ilíacas. Analisamos as correlações entre o comprimento ureteral e as demais medidas antropométricas. Resultados: Foram dissecados os ureteres de 115 cadáveres adultos no período entre abril/2009 e janeiro/2012. O comprimento ureteral médio não variou o gênero, nem com a estatura. Não foi encontrada correlação entre o comprimento ureteral e as medidas antropométricas pesquisadas na população geral analisada, bem como nos subgrupos analisados. Não se evidenciou diferenças significantes entre as medidas dos ureteres direito e esquerdo. Conclusões: Não há diferença no comprimento ureteral médio entre as diferentes faixas de altura e entre os gêneros masculino e feminino. Não há correlação significante entre o comprimento ureteral e as demais medidas antropométricas.
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Mechanisms of epithelial branching, nephrogenesis, and the role of the Rho-GTPase family in kidney developmentLindström, Nils Olof January 2009 (has links)
The metanephric kidney consists of two types of epithelia; the Wolffian duct-derived ureteric bud and the nephrogenic components that originate from mesenchymal-toepithelial transitions in the metanephric mesenchyme. The ureteric bud forms when inductive signals from the metanephric mesenchyme stimulates the evagination of an epithelial tube from the Wolffian duct into the mesenchyme. Reciprocal signalling between the ureteric bud and the metanephric mesenchyme regulates the branching of the ureteric bud and the induction of nephron formation. Inductive and inhibitory signalling of ureteric bud growth and branching has been shown by several protein families, however, the mechanical aspects of ureteric bud branching and nephrogenesis are largely unknown. I investigated the roles of Rac1-GTPase and Rho-kinase during kidney development. These proteins are important regulators of the cytoskeleton where Rac1 is a promoter of actin filament polymerisation and Rho-kinase directly stimulates the formation and contraction of actin-myosin stress fibres. Using a cell-permeable inhibitor, Rac1 was inhibited with no effects on nephron formation or subsequent segmentation and patterning. Inhibition of active Rac1 significantly reduced the level of ureteric bud branching and also resulted in lower proliferation rates. Rho-kinase was similarly targeted using two inhibitors. Rho-kinase inhibition had important effects on nephron formation and nephron maturation. Inhibition of Rhokinase resulted in decreased levels of nephron formation and severely morphologically abnormal nephrons. The formation of apical-basal polarity was disturbed as was the development of the visceral and parietal epithelia; precursors of the renal corpuscle. Inhibition of Rho-kinase led to abnormal formation of the proximal-distal axis and abnormal segmentation of the nephron. The effects of Rho-kinase inhibition were partially mimicked by direct targeting of actin-myosin contractions using a myosin-ATPase inhibitor. This demonstrated that Rho-kinase is necessary during multiple stages of nephrogenesis and maturation, at least in part, as a result of its ability to regulate actin-myosin contraction. These results show that Rac1 and Rho-kinase play important roles during several aspects of kidney development and highlights the significance of further investigating the mechanisms involved during kidney organogenesis.
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Role of Tshz3 in the development and function of the kidney / Rôle de Tshz3 dans le développement et le fonctionnement du reinSanchez Martin, Irene 18 December 2018 (has links)
Les anomalies du tractus rénal et les troubles du spectre autistique caractérisent le syndrome 19q12 causé par la délétion hétérozygote du gène TSHZ3. Pour identifier des programmes développementaux TSHZ3-dépendants, nous avons comparé le transcriptome (RNA-seq) d’uretères et de reins mutants Tshz3 (KO) et sauvage (WT) d’embryons de souris au stade E12,5. Nous avons identifié des gènes exprimés de façon différentielle connus pour être impliqués dans le développement de l'uretère et/ou des reins, dont 38 ont des orthologues humains associés à des maladies rénales. Corrélativement, les reins E12,5 KO présentent une arborisation anormale de l’uretère.Dans les reins adultes, TSHZ3 est exprimé dans les cellules endothéliales glomérulaires. L'analyse morphologique de reins Tshz3+/lacZ (HET) révèle une diminution de la densité des glomérules et de l'épaisseur de la membrane basale glomérulaire ainsi qu'un phénotype d'effacement des pieds des podocytes. L’analyse du sang et de l'urine de souris adultes HET a permis d’établir des profils spécifiquement associés au génotype HET. En particulier, le protéome urinaire a identifié 33 biomarqueurs qui pourraient constituer la signature d'un processus pathologique. Par ailleurs, l'analyse du transcriptome des reins HET adultes montre un enrichissement pour des voies liées à l'inflammation.Ces résultats confirment le rôle précoce de Tshz3 dans l'uretère ainsi qu'une fonction de Tshz3 dans les reins embryonnaires. La présence de défauts structurels et fonctionnels dans les reins hétérozygotes adultes Tshz3 renforce l'idée que les souris HET modélisent le syndrome TSHZ3 humain. / Renal tract defects and autism spectrum disorder represent the phenotypic core of the 19q12 syndrome caused by heterozygote deletion of the TSHZ3 gene. To identify TSHZ3-dependent developmental programs, we performed a transcriptome analysis (RNA-seq) on E12.5 Tshz3 mutants (KO) and wild type (WT) control mouse ureters and kidneys. This analysis identified differentially expressed genes known to be involved in ureter and/or kidney development, among which 38 have human orthologues associated to renal tract diseases. Correlatively, we found that E12.5 Tshz3 KO kidneys display an abnormal ureteric branching morphogenesis.In adult kidneys, we showed that TSHZ3 is expressed in glomerular endothelial cells. Histological and transmission electron microscopy analysis showed a decreased glomerular density and thickness of the glomerular basement membrane as well as a foot process effacement phenotype in Tshz3+/lacZ (HET) kidneys. To evaluate renal function, we analysed blood and urine samples from HET and WT adult mice. Both analyses generated profiles that specifically associated with HET genotype. In particular, the urine proteome identified 33 biomarkers that might constitute a signature for a pathological process in HET kidneys. Note that transcriptome analysis of adult HET kidneys showed enrichment for inflammation-related pathways.These results support an early role for Tshz3 in the ureter as well as an unanticipated function for Tshz3 in E12.5 embryonic kidneys. The presence of structural and functional defects in Tshz3 heterozygous adult kidneys reinforces the idea that HET mice model the human TSHZ3 disorder.
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Host Responses to Infection of the Upper and Lower Urinary TractBowen, Samantha January 2013 (has links)
<p>Urinary tract infections (UTIs) are the second most common type of infection identified in the clinical setting and disproportionately afflict women. UTIs most frequently manifest in the form of infection of the lower urinary tract, involving the bladder. Uropathogens, particularly uropathogenic E. coli, progressively colonize the urethra and ascend to the bladder, where they initiate cystitis. In some cases, infection further ascends through the ureters and reaches the kidneys, where it causes pyelonephritis. Infection of both the upper and lower urinary tract can have serious ramifications for the host, and this is in large part due not to infection itself but to host-directed responses to bacterial insults. </p><p> In this thesis, I will describe and discuss two distinct aspects of UTIs. In the first study, in vivo work in a mouse model of urinary tract infection revealed a novel role for mast cells, which are tissue-resident granulated innate immune cells, in directing the detachment and death of epithelial cells during cystitis, facilitating the clearance of bacteria from the bladder. An ex vivo porcine bladder infection model suggested a specific role for mast cell granules and the proteases contained therein, which was corroborated with in vitro experiments utlizing isolated mast cell granules and human epithelial cells to demonstrate granule-induced exfoliation and cell death. From this work, it is clear that mast cells play a highly targeted role in modulating urothelial integrity during bladder infection by mediating host-directed epithelial loss.</p><p> In the second study described in this dissertation, the synergistic roles of both pyelonephritis and vesico-ureteric reflux (VUR), a congenital urinary tract defect that results in the improper backflow of urine from the bladder to the kidney, in the development of reflux nephropathy, a fibrotic host response characterized by renal scar formation, were elucidated in a series of in vivo experiments. Specifically, the C3H mouse, which is naturally susceptible to VUR, was utilized to characterize the dynamics of kidney infection and the onset of reflux nephropathy. Renal scarring was dependent on the presence of sustained kidney infection and the accompanying inflammatory response due to VUR, while neither transient infection nor reflux alone were sufficient to provoke nephropathy. Thus, the development of reflux nephropathy is dependent upon the confluence of both infection and VUR. </p><p> This body of work reveals the double-edged sword of the host inflammatory response to urinary tract infection. In the bladder, mast cell activation and degranulation leads to granule-induced epithelial exfoliation and consequently a reduction in the bacterial burden in the bladder. However, the sustained inflammatory response that accompanies pyelonephritis in vesico-ureteric reflux-affected individuals results in significant damage to the kidney without any accompanying reduction in infection. These findings highlight the dueling roles of the host inflammatory response to infection in the upper and lower urinary tract and strongly suggest that differential clinical approaches to cystitis and pyelonephritis are necessary to promote an effective mast cell in the bladder in the former and facilitate the clearance of renal infection while mitigating tissue damage in the latter.</p> / Dissertation
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Expansion of human iPSC-derived ureteric bud organoids with repeated branching potential / 繰り返す分岐形態形成能力を有するヒトiPS細胞由来尿管芽オルガノイドの作製と拡大培養Ryosaka, Makoto 25 January 2021 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22879号 / 医博第4673号 / 新制||医||1047(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 川口 義弥, 教授 柳田 素子, 教授 小川 修 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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Exogenous modulation of embryonic tissue and stem cells to form nephronal structuresSebinger, David Daniel Raphael 04 July 2013 (has links) (PDF)
Renal tissue engineering and regenerative medicine represent a significant clinical objective because of the very limited prospect of cure after classical kidney treatment. Thus, approaches to isolate, manipulate and reintegrate structures or stimulating the selfregenerative potential of renal tissue are of special interest. Such new strategies go back to knowledge and further outcome of developmental biological research. An understanding of extracellular matrix (ECM) structure and composition forms thereby a particularly significant aspect in comprehending the complex dynamics of tissue regeneration. Consequently the reconstruction of these structures offers beneficial options for advanced cell and tissue culture technology and tissue engineering. In an effort to investigate the influence of natural extracellular structures and components on embryonic stem cell and renal embryonic tissue, methodologies which allow the easy application of exogenous signals on tissue in vitro on the one hand and the straight forward evaluation of decellularization methods on the other hand, were developed. Both systems can be used to investigate and modulate behaviour of biological systems and represent novel interesting tools for tissue engineering. The novel technique for culturing tissue in vitro allows the growing of embryonic renal explants in very low volumes of medium and optimized observability, which makes it predestined for testing additives. In particular, this novel culture set up provides an ideal opportunity to investigate renal development and structure formation. Further studies indicated that the set is universally applicable on all kinds of (embryonic) tissue. Following hereon, more than 20 different ECM components were tested for their impact on kidney development under 116 different culture conditions, including different concentrations and being either bound to the substrate or dissolved in the culture medium. This allowed to study the role of ECM constituents on renal structure formation. In ongoing projects, kidney rudiments are exposed to aligned matrix fibrils and hydrogels with first promising results. The insights gained thereof gave rise to a basis for the rational application of exogenous signals in regenerative kidney therapies. Additionally new strategies for decellularization of whole murine adult kidneys were explored by applying different chemical agents. The obtained whole matrices were analysed for their degree of decellularization and their residual content and composition. In a new straight forward approach, a dependency of ECM decellularization efficiency to the different agents used for decellularization could be shown. Moreover the capability of the ECM isolated from whole adult kidneys to direct stem cell differentiation towards renal cell linage phenotypes was proved. The data obtained within this thesis give an innovative impetus to the design of biomaterial scaffolds with defined and distinct properties, offering exciting options for tissue engineering and regenerative kidney therapies by exogenous cues.
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Exogenous modulation of embryonic tissue and stem cells to form nephronal structuresSebinger, David Daniel Raphael 26 April 2013 (has links)
Renal tissue engineering and regenerative medicine represent a significant clinical objective because of the very limited prospect of cure after classical kidney treatment. Thus, approaches to isolate, manipulate and reintegrate structures or stimulating the selfregenerative potential of renal tissue are of special interest. Such new strategies go back to knowledge and further outcome of developmental biological research. An understanding of extracellular matrix (ECM) structure and composition forms thereby a particularly significant aspect in comprehending the complex dynamics of tissue regeneration. Consequently the reconstruction of these structures offers beneficial options for advanced cell and tissue culture technology and tissue engineering. In an effort to investigate the influence of natural extracellular structures and components on embryonic stem cell and renal embryonic tissue, methodologies which allow the easy application of exogenous signals on tissue in vitro on the one hand and the straight forward evaluation of decellularization methods on the other hand, were developed. Both systems can be used to investigate and modulate behaviour of biological systems and represent novel interesting tools for tissue engineering. The novel technique for culturing tissue in vitro allows the growing of embryonic renal explants in very low volumes of medium and optimized observability, which makes it predestined for testing additives. In particular, this novel culture set up provides an ideal opportunity to investigate renal development and structure formation. Further studies indicated that the set is universally applicable on all kinds of (embryonic) tissue. Following hereon, more than 20 different ECM components were tested for their impact on kidney development under 116 different culture conditions, including different concentrations and being either bound to the substrate or dissolved in the culture medium. This allowed to study the role of ECM constituents on renal structure formation. In ongoing projects, kidney rudiments are exposed to aligned matrix fibrils and hydrogels with first promising results. The insights gained thereof gave rise to a basis for the rational application of exogenous signals in regenerative kidney therapies. Additionally new strategies for decellularization of whole murine adult kidneys were explored by applying different chemical agents. The obtained whole matrices were analysed for their degree of decellularization and their residual content and composition. In a new straight forward approach, a dependency of ECM decellularization efficiency to the different agents used for decellularization could be shown. Moreover the capability of the ECM isolated from whole adult kidneys to direct stem cell differentiation towards renal cell linage phenotypes was proved. The data obtained within this thesis give an innovative impetus to the design of biomaterial scaffolds with defined and distinct properties, offering exciting options for tissue engineering and regenerative kidney therapies by exogenous cues.:Table of Contents
LISTS OF FIGURES AND TABLES VI
ACKNOWLEDGEMENTS..................................................................................VII
ABSTRACT ............................................................................................................IX
NOMENCLATURE ................................................................................................X
1 INTRODUCTION...................................................................................................1
2 FUNDAMENTALS..................................................................................................2
2.1 KIDNEY DEVELOPMENT AND REGENERATION ...............................................................................2
2.1.1 Function of the kidney............................................................................................2
2.1.2 Development of the metanephric kidney ................................................................2
2.1.3 Selfregenerative potential of the kidney.................................................................5
2.2 THE EXTRACELLULAR MATRIX AS BIOLOGICAL SCAFFOLD ...............................................................6
2.2.1 Molecular composition of the ECM........................................................................7
2.2.1.1 An overview of the main ECM components..................................................................................8
2.2.2 Cell/tissue-matrix interactions.............................................................................12
2.2.2.1 Biochemical signals....................................................................................................................13
2.2.2.2 Mechanical signals......................................................................................................................14
2.2.2.3 Structural signals........................................................................................................................15
2.3 TISSUE ENGINEERING FOR THERAPEUTIC PURPOSES .....................................................................15
2.3.1 An overview of tissue engineering and regenerative medicine.............................15
2.3.2 Biomaterials for tissue engineering and regenerative medicine...........................18
2.3.2.1 Decellularization approach as tool to extract natural matrices....................................................19
2.3.3 Tissue engineering and regenerative medicine in kidney treatment.....................19
2.4 ORGAN AND TISSUE CULTURE AS TOOL FOR TISSUE ENGINEERING...................................................22
2.4.1 Common organ culture systems............................................................................24
3 OBJECTIVES AND MOTIVATION...................................................................25
4 RESULTS AND DISCUSSION............................................................................27
4.1 A NOVEL, LOW-VOLUME METHOD FOR ORGAN CULTURE OF EMBRYONIC KIDNEYS THAT ALLOWS
DEVELOPMENT OF CORTICO-MEDULLARY ANATOMICAL ORGANIZATION..............................................27
4.1.1 Additional evidences (to Appendix A) for stress reduction of kidney rudiments
cultured in the novel system than those grown in conventional organ culture.....28
4.1.2 Additional evidences (to Appendix A) for corticomedullary zonation and improved
development of kidney rudiments cultured in the novel system for a period of 12
days......................................................................................................................30
4.1.3 Additional evidences (to Appendix A) for the application of the glass based low
volume culture system for other organs................................................................32
4.2 ECM MODULATED EARLY KIDNEY DEVELOPMENT IN ORGAN CULTURE ...........................................34
4.3 ESTABLISHING AND EVALUATING DECELLULARIZATION TECHNIQUES TO ISOLATE WHOLE KIDNEY ECMS
FROM ADULT MURINE KIDNEYS................................................................................................37
4.4 THE ABILITY OF WHOLE DECELLULARIZED ECM CONSTRUCTS TO INFLUENCE MURINE EMBRYONIC STEM
CELL DIFFERENTIATION AND RENAL TISSUE BEHAVIOUR IN A NEW STRAIGHT FORWARD APPROACH..........38
iv
5 SUMMARY AND OUTLOOK.............................................................................39
5.1 SUMMARY..........................................................................................................................39
5.2 OUTLOOK...........................................................................................................................42
6 BIBLIOGRAPHY.................................................................................................49
7 APPENDICES..........................................................................................................I
7.1 APPENDIX A: A NOVEL, LOW-VOLUME METHOD FOR ORGAN CULTURE OF EMBRYONIC KIDNEYS
THAT ALLOWS DEVELOPMENT OF CORTICO-MEDULLARY ANATOMICAL ORGANIZATION......................I
7.2 APPENDIX B: ECM MODULATED EARLY KIDNEY DEVELOPMENT IN EMBRYONIC ORGAN CULTURE ....XIX
7.3 APPENDIX C: THE DEWAXED ECM: AN EASY METHOD TO ANALYZE CELL BEHAVIOUR ON
DECELLULARIZED EXTRACELLULAR MATRICES.......................................................................XLIV
7.4 PUBLICATIONS AND SCIENTIFIC CONTRIBUTIONS......................................................................LXV
7.5 SELBSTSTÄNDIGKEITSERKLÄRUNG......................................................................................LXIX
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