A complacência da bexiga depende de músculos lisos, fibras colágenas, fibras elásiticas e suas relações. O objetivo deste trabalho é determinar a composição da matriz extracelular em amostras de bexigas normais através de análise bioquímica de colágeno e glicosaminoglicanos em amostras obtidas de mulheres em diferentes grupos de idade, analisando separadamente as camadas urotelial e muscular. Avaliamos 17 amostras de bexiga divididas em três grupos: infância (N=5), menacme (N=6) e pós-menopausa (N=6). As bexigas foram analisadas para concentração de GAG total e colágeno e para análise qualitativa de GAG por eletroforese em gel de agarose. Na camada muscular, não houve diferença entre os grupos tanto para GAG quanto para colágeno. Na camada urotelial, a análise da concentração de colágeno não mostrou diferença entre os grupos, mas a concentração de GAG no grupo da pós-menopausa (0.21 0.12 μg de ácido hexurônico/mg de tecido seco) apresentou diferença em relação aos grupos do menacme (1.78 1.62 μg de ácido hexurônico/mg de tecido seco) e da infância ( 2.29 1.32 μg de ácido hexurônico/mg de tecido seco).Nosso trabalho concluiu que a concentração de GAG está substancialmente diminuída na camada urotelial da bexiga de mulheres na pós-menopausa. / Bladder compliance is dependent on smooth muscle, collagen fibers, elastic fiber and their ratios. The luminal surface of the urothelium is covered by an adhering glycosaminoglycan (GAG) layer. The aim of this study was to determine the composition of the extracellular matrix (ECM) in normal samples of women bladders through biochemistry analysis of collagen and GAG on samples obtained from individuals from different age groups, analyzing separately the urothelial and muscular layers. We studied samples taken from bladders of 17 patients divided in three different groups: childhood (N=5), menacme (N=6) and menopause (N=6). Bladders were analyzed for total GAG and collagen concentration per mg dry tissue and for the contents of GAG species, as determined by agarose electrophoresis and reported as the percent of total sulfated GAG. In muscular layer, collagen and GAG concentration showed no difference between groups. In urothelial layer, collagen concentration showed no difference between groups but GAG concentration in menopause ( 0.21 0.12 μg hexuronic acid /mg dry tissue) was different from menacme (1.78 1.62 μg hexuronic acid /mg dry tissue) and childhood ( 2.29 1.32 μg hexuronic acid /mg dry tissue). There was no difference between sulfated GAG in three groups.In conclusion, GAG concentration in urothelial layer was substantially lower in menopause women.

It is a well established phenomenon that the epithelial lining of the urinary bladder (urothelium) when implanted into skeletal muscle induces ectopic ossification. However, despite numerous observations, this reaction is poorly understood. This research further studied this reaction by - (a) demonstrating the reaction in a suitable small animal model; (b) attempting to induce the reaction by implanting urothelial cells purified by cell culture techniques; and (c) comparing the bone forming reaction induced by implanted urothelium to the reaction induced by implanting Bone Marrow Stem Cells (BMSC's) and Osteophyte Stem Cells (OSC's). By demonstrating newly formed bone after the implantation of guinea pig urothelium into the skeletal muscle of a Severe Combined Immuno-Deficient Mouse (SCID-Mouse) this research demonstrated that a suitable small animal model had been established. This is despite inherent difficulties (particularly bacterial contamination) associated with establishing a primary cell culture of guinea pig urothelial cells. Additionally, the intramuscular ectopic osteoinductive potential of human BMSC's (hBMSC's) in the SCID-mouse has also been demonstrated. Confirming that the injection of cultured cells in suspension is an adequate intramuscular delivery technique, this research demonstrates that hBMSC's induce ectopic ossification by non-immunological means. This research has demonstrated a number of differences between urothelium induced ectopic ossification and ectopic ossification induced by BMSC's, suggesting they are two separate processes. This is important because the chemotaxis and subsequent osteogenic differentiation of BMSC's has previously been one of the more popular postulated mechanisms of urothelium induced ectopic ossification. Finally, this research has demonstrated the ectopic osteoinductive potential of stem cells isolated from the marrow of human osteophytes (human Osteophyte Stem Cells, hOSC's). This observation has not been previously reported, and will hopefully provide a valuable contribution to a body of knowledge that has important ramifications in both the treatment of osteoarthritis, and the use of BMSC's in tissue engineering.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 3 uppsatser.

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No. of bitstreams: 1 vasconcellos_al_dr_jabo.pdf: 2492463 bytes, checksum: 31f8926ecfbe121c3aaf71eb9b3e0bf5 (MD5) Previous issue date: 2016-07-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O objetivo do presente estudo foi padronizar a metodologia de citologia de lavado vesical preparado pelo método de citocentrifugação, avaliar o padrão das células epiteliais de transição em cães e aplicabilidade clínica da citopatologia esfoliativa como método complementar de diagnóstico de doenças vesicais em cães. O estudo foi realizado em duas fases. A primeira incluiu 34 pacientes sem anormalidade ou doença vesical (cães adultos, machos e fêmeas), para padronização da metodologia e caracterização da celularidade normal. Na segunda fase foram avaliados 95 pacientes (cães adultos, machos e fêmeas), com ou sem sinais do trato urinário inferior. Em ambas as fases os cães foram avaliados por meio de exames de rotina, com destaque para sedimentoscopia de urina, urocultura e exame ultrassonográfico da bexiga, além da citologia de lavado vesical preparada pelo método de citocentrifugação (colorações de Papanicolaou, Giemsa, Rosenfeld modificado e Gram – somente na segunda fase). Os resultados obtidos indicaram que a técnica empregada no estudo para recuperação celular realizada com barbotagens de três ciclos utilizando 1mL/kg de solução salina, se mostrou eficaz para avaliação de células uroteliais de cães sadios. Com o segundo estudo, que incluiu pacientes com doença vesical, concluiu-se que o conjunto de avaliações para o diagnóstico de doença vesical empregado são complementares e as contribuições de cada exame variam de acordo com o tipo de enfermidade a ser diagnosticada. A citologia de lavado vesical se mostrou como exame tão relevante quanto a sedimentoscopia no que se refere às características de sensibilidade e especificidade para detectar sadios e doentes, mas com as vantagens de conferir refinamento aos diagnósticos ou viabilizar o diagnóstico não concluído pelos exames anteriores. / The objective of this study was to standardize the methodology for cytology bladder washing prepared by cytospin method to evaluate the pattern of epithelial transition cells in dogs and the clinical applicability of exfoliative cytology as a complementary method for of bladder diseases diagnosis in dogs. The study was conducted in two phases. The first one included 34 patients without bladder abnormality or disease (adult dogs, male and female), for standardization of the methodology and characterization of the typical cellularity. In the second phase it was evaluated a group of 95 patients (adult dogs, male and female), with or without lower urinary tract signs. In both phases the dogs were evaluated by routine tests, especially sediment of urine, urine culture and ultrasound bladder examination, as well as cytology of bladder washing prepared by cytospin (stains of Papanicolaou, Giemsa, Rosenfeld modified and Gram - only in the second phase). The results indicated that the technique used in the study for cell recovery, performed with three barbotage cycles using 1 mL/kg of saline, was effective for urothelial cells evaluation from healthy dogs. In the second study, which included patients with bladder disease, it is concluded that the employed set of evaluations for bladder disease diagnosis are complementary and the contributions of each test vary with the type of disease to be diagnosed. The cytology of bladder washing showed was as relevant as the sediment, considering the sensitivity and specificity to detect healthy and sick patients, but with the advantages of giving refinement to enable diagnosis or the diagnosis not completed by previous tests.

A complacência da bexiga depende de músculos lisos, fibras colágenas, fibras elásiticas e suas relações. O objetivo deste trabalho é determinar a composição da matriz extracelular em amostras de bexigas normais através de análise bioquímica de colágeno e glicosaminoglicanos em amostras obtidas de mulheres em diferentes grupos de idade, analisando separadamente as camadas urotelial e muscular. Avaliamos 17 amostras de bexiga divididas em três grupos: infância (N=5), menacme (N=6) e pós-menopausa (N=6). As bexigas foram analisadas para concentração de GAG total e colágeno e para análise qualitativa de GAG por eletroforese em gel de agarose. Na camada muscular, não houve diferença entre os grupos tanto para GAG quanto para colágeno. Na camada urotelial, a análise da concentração de colágeno não mostrou diferença entre os grupos, mas a concentração de GAG no grupo da pós-menopausa (0.21 0.12 μg de ácido hexurônico/mg de tecido seco) apresentou diferença em relação aos grupos do menacme (1.78 1.62 μg de ácido hexurônico/mg de tecido seco) e da infância ( 2.29 1.32 μg de ácido hexurônico/mg de tecido seco).Nosso trabalho concluiu que a concentração de GAG está substancialmente diminuída na camada urotelial da bexiga de mulheres na pós-menopausa. / Bladder compliance is dependent on smooth muscle, collagen fibers, elastic fiber and their ratios. The luminal surface of the urothelium is covered by an adhering glycosaminoglycan (GAG) layer. The aim of this study was to determine the composition of the extracellular matrix (ECM) in normal samples of women bladders through biochemistry analysis of collagen and GAG on samples obtained from individuals from different age groups, analyzing separately the urothelial and muscular layers. We studied samples taken from bladders of 17 patients divided in three different groups: childhood (N=5), menacme (N=6) and menopause (N=6). Bladders were analyzed for total GAG and collagen concentration per mg dry tissue and for the contents of GAG species, as determined by agarose electrophoresis and reported as the percent of total sulfated GAG. In muscular layer, collagen and GAG concentration showed no difference between groups. In urothelial layer, collagen concentration showed no difference between groups but GAG concentration in menopause ( 0.21 0.12 μg hexuronic acid /mg dry tissue) was different from menacme (1.78 1.62 μg hexuronic acid /mg dry tissue) and childhood ( 2.29 1.32 μg hexuronic acid /mg dry tissue). There was no difference between sulfated GAG in three groups.In conclusion, GAG concentration in urothelial layer was substantially lower in menopause women.

No description available.

Background: The success of kidney transplant surgery and ureteral reconstruction requires the preservation of the ureteral blood supply. Because of its potential vulnerability to surgical trauma during trans plant and reconstructive surgery, the ureteral vasculature merits a full anatomical description. Methods: The microvascular anatomy of the ureter was studied in male New Zealand white rabbits by light microscopy and transmission electron microscopy and scanning electron microscopy of vascular corrosion casts and alkali digested tissue. Results: The rabbit ureter is supplied predominantly by a branch of the renal artery proximally (cranial ureteral artery) and by a branch of the vesicular artery distally (caudal ureteral artery). Minor vascular continuities are also present between the capillary beds of the ureter and those of the renal pelvis cranially and the bladder wall caudally. There are no external vascular connections to the middle ureter with the exception of a single, small vein which drains into the inferior vena cava. A single group of longitudinal arteries and veins runs the full length of the ureter within the adventitia. Branches of these longitudinal vessels pass tangentially through the muscularis to supply a vascular complex within the lamina propria. This complex in turn supports a rich, mucosal capillary plexus located at the junction between the transitional epithelium and the lamina propria. In the fixed ureter the capillary plexus lies in grooves formed by displacement of the basal layers of the overlying transitional epithelium. The capillaries are continuous or fenestrated, are often invested with pericytes, and are distributed uniformly around the entire circumference of the ureter. Conclusions: The ureteral vasculature exhibits several unique features related to its function in urine conduction and its ability to accommodate expansion and contraction. The combination of techniques used provides a clear three‐dimensional view of this vasculature. Our findings also confirm that, because of its limited blood supply, the ureter may be very susceptible to injury during renal transplantation or other abdominal surgery.

No description available.

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.

Objectives: The mucosa of human detrusor strips impairs catecholamine-induced relaxation. In order to elucidate which signal transduction pathways are involved in this cross talk between the mucosa and detrusor, we have studied the effects of several pharmacological agonists and antagonists on noradrenaline-mediated relaxation in intact and mucosa-denuded detrusor strips. Patients and Methods: Strips of detrusor tissue were obtained from patients who had undergone cystectomy for bladder cancer and were set up for force measurement. KCl- or carbachol-precontracted strips were relaxed with increasing concentrations of noradrenaline in the absence and in the presence of nitric oxide synthase inhibitor, L-NAME; P2X-receptor antagonist, PPADS; ET A -receptor antagonist, BQ-123; ET B -receptor antagonist, BQ-788; cyclooxygenase inhibitor, diclofenac; AT 1 -receptor antagonist, candesartan; and NK 1 -receptor antagonist, L-703,606. Results: In intact strips, KCl-stimulated force was enhanced by all blockers; carbachol-stimulated force increased with L-703,606. In denuded strips, only L-NAME augmented the KCl-stimulated contraction. Noradrenaline relaxed the precontracted detrusor strips to a significantly larger extent and at lower concentrations in denuded than in intact strips. L-NAME, PPADS and BQ-123/BQ-788 had little effect on noradrenaline-induced relaxation, whereas diclofenac, candesartan and L-703,606 sensitized intact carbachol-stimulated detrusor strips to noradrenaline-induced relaxation. Conclusion: Inhibition of the noradrenaline-induced relaxation of precontracted human detrusor strips by the mucosa is attenuated by diclofenac, candesartan and L-703,606 suggesting the involvement of prostanoids, angiotensin and neurokinin pathways. Further experiments are required to unravel the exact mechanisms.