Urethroplasty Using Autologous Urethral Tissue-embedded Acellular Porcine Bladder Submucosa Matrix Grafts for the Management of Long-Segment Urethral Stricture in a Rabbit Model

We conducted this study to evaluate the combined effect of acellular bladder submucosa matrix (BSM) and autologous urethral tissue for the treatment of long segment urethral stricture in a rabbit model. To prepare the BSM, porcine bladder submucosa was processed, decellularized, configured into a sheet-like shape, and sterilized. Twenty rabbits were randomized to normal control, urethral stricture, urethroplasty using BSM only or BSM/autologous urethral tissue (n=5 per group). Retrograde urethrography was performed at 4, 8, and 12 weeks postoperatively, and the grafted specimens were harvested at week 12 to evaluate urethral reconstruction through histopathologic and immunohistochemical analysis. The mean urethral width of the control, stricture, BSM, and BSM/autologous urethral tissue groups at week 12 was 10.3+/-0.80, 3.8+/-1.35, 8.8+/-0.84, and 9.1+/-1.14 mm, respectively. The histopathologic study revealed that the BSM/autologous urethral tissue graft had a normal area of urethral lumen, compact muscular layers, complete epithelialization, and progressive infiltration by vessels in the regenerated urethra. In contrast, the BSM grafts revealed keratinized epithelium, abundant collagenized fibrous connective tissue, and were devoid of bundles of circular smooth muscle. Nontransected ventral onlay-augmented urethroplasty using an acellular BSM scaffold combined with an autologous urethral tissue graft represents a feasible procedure for urethral reconstruction.

Urethroplasty Using Autologous Urethral Tissue-embedded Acellular Porcine Bladder Submucosa Matrix Grafts for the Management of Long-Segment Urethral Stricture in a Rabbit Model

We conducted this study to evaluate the combined effect of acellular bladder submucosa matrix (BSM) and autologous urethral tissue for the treatment of long segment urethral stricture in a rabbit model. To prepare the BSM, porcine bladder submucosa was processed, decellularized, configured into a sheet-like shape, and sterilized. Twenty rabbits were randomized to normal control, urethral stricture, urethroplasty using BSM only or BSM/autologous urethral tissue (n=5 per group). Retrograde urethrography was performed at 4, 8, and 12 weeks postoperatively, and the grafted specimens were harvested at week 12 to evaluate urethral reconstruction through histopathologic and immunohistochemical analysis. The mean urethral width of the control, stricture, BSM, and BSM/autologous urethral tissue groups at week 12 was 10.3+/-0.80, 3.8+/-1.35, 8.8+/-0.84, and 9.1+/-1.14 mm, respectively. The histopathologic study revealed that the BSM/autologous urethral tissue graft had a normal area of urethral lumen, compact muscular layers, complete epithelialization, and progressive infiltration by vessels in the regenerated urethra. In contrast, the BSM grafts revealed keratinized epithelium, abundant collagenized fibrous connective tissue, and were devoid of bundles of circular smooth muscle. Nontransected ventral onlay-augmented urethroplasty using an acellular BSM scaffold combined with an autologous urethral tissue graft represents a feasible procedure for urethral reconstruction.

Analysis of the reactivity of indirect immunofluorescence in patients with pemphigus foliaceus and pemphigus vulgaris using rat bladder epithelium as a substrate

OBJECTIVES: To evaluate the reactivity of indirect immunofluorescence using rat bladder epithelium as a substrate in patients with pemphigus foliaceus and pemphigus vulgaris from the Department of Dermatology, University of São Paulo Medical School, Brazil. METHODS: Thirty-two patients (8 male and 24 female) from the Department of Dermatology, University of São Paulo Medical School, were selected. Three had mucosal pemphigus vulgaris, 20 had mucocutaneous pemphigus vulgaris, and 9 had pemphigus foliaceus. Patients’ sera were tested by indirect immunofluorescence performed on human foreskin and rat bladder epithelium and by ELISA assays utilizing baculovirus-expressed recombinant desmoglein 3 and desmoglein 1. RESULTS: No patients with mucosal pemphigus vulgaris, 5 of 20 patients with mucocutaneous pemphigus vulgaris (25 percent) and 4 of 9 patients with pemphigus foliaceus (44 percent) had positive indirect immunofluorescence using rat bladder epithelium as a substrate. CONCLUSION: Indirect immunofluorescence using rat bladder epithelium as a substrate is recommended whenever a diagnosis of paraneoplastic pemphigus is considered. The identification of a subset of pemphigus foliaceus and pemphigus vulgaris patients that recognizes desmoplakins by this laboratory tool is critical to avoid the misdiagnosis of paraneoplastic pemphigus.

effectiveness of intravesical Botox in idiopathic and non-idiopathic detrusor over activity

We have evaluated and compared the efficacy of Botox[registered] in the treatment of bladder over activity in Idiopathic detrusor over activity [IDO] and Neuropathic detrusor over activity [NDO] in a tertiary centre. We hypothesised that the outcome of this less invasive treatment would be more promising in patients with IDO than those with NDO. A total of 229 patients were reviewed with overactive bladder proven by urodynamic studies over the last 2 years. Of these, 174 patients received Botox[registered] injection. 132 [75%] patients had IDO and 42 [25%] patients had NDO. 200 IU of Botox[registered] was injected per patient in the IDO group and 300 IU in the neurogenic group. Eighty one of 118 [68.7%] known patients with IDO reported being dry after receiving the Botox[registered] compared to 20/39 [51.3%] patients with NDO. Whereas, 88/118 [68.7%] patients with IDO reported significant satisfaction compared to 25/39 [64.1%] patients with NDO. Botox[registered] is an effective, minimally invasive treatment for drug refractory IDO+NDO. In comparison, patient reported satisfaction and dryness with the procedure was statistically the same. However, the IDO group responded slightly better to treatment. The average mean effect was the same in both groups

Estudio del sistema purinérgico a través de la caracterización de la enzima E-NPP3 y de la secreción de ATP en células uroepiteliales de la vejiga urinaria del ratón / Study of the purinergic system through the characterization of the E-NPP3 enzyme and the secretion of ATP in uroepithelial cells of the urinary bladder of the mouse

El presente trabajo involucra el estudio del sistema purinérgico mediante la purinérgico mediante la purificación y la caracterización bioquimica de la ecto-enzima E-NPP3 soluble, la cual modula la activación de los receptores purinérgicos mediante la hidrólisis de los nucleótidos extracelulares. La purificación de esta enzima, expresada en la línea celular CHO-K1, requirió el empleo de diferentes columnas cromatográficas; comprobándose la pureza, mediante la determinación de la actividad enzimática, la cuantificación de las proteínas y la detección de dicha enzima. Los resultados de la caracterización de la E-NPP3 indican que presentan un ph óptimo alcalino y que su actividad depende de la la concentración de los iones calcio y magnesio: mientras que el imidazol y el DDT ejercen acciones inhibidoras sobre su actividad. La purificación de la enzima E-NPP3 soluble representa un primer paso para futuros estudios que permitan su cristalización lo cual, constituirá una herramienta en la elaboración de agonistas y antagonistas selectivos o de anticuerpos monoclonales contra dicha enzima, útiles en el diagnóstico o el tratamiento de condiciones patológicas como el cáncer de colon y la colangiocarcinoma. Adicionalmente, teniendo en cuenta que se ha demostrado la disminución de la expresión de las ecto-nucleotidasas en el uroepitelio de pacientes que padecen cistitis intersticial; el presente trabajo comprende también, el desarrollo de un modelo de órgano aislado, la vejiga urinaria del ratón, para el estudio de la secreción de ATP desde las células uroepiteliales. Dicho modelo permitió demostrar, mediante estudios electrofisiológicos y el uso de diferentes fármacos, la contriución de los receptores purinérgicos sobre la actividad eléctrica del nervio pélvico cuando la vejiga es sometida a distensión mecánica gradual; sugiriendo la importancia de estos receptores en la transducción mecanosensorial de dicho órgano y permitiendo inferir la posible interrelación con los receptores de vaniloides en la detección de los estímulos sensoriales por parte del uroepitelio

c-fos Expression in Bladder-Specific Spinal Neurons after Spinal Cord Injury Using Pseudorabies Virus

PURPOSE: c-fos expression in spinal neurons that are activated by lower urinary tract stimulation are not organ specific. In this experiment, we demonstrated changes of c-fos expression in bladder-specific preganglionic neurons (PGNs) and interneurons using pseudorabies virus (PRV). MATERIALS AND METHODS: Forty Sprague-Dawley rats were used. We identified the neuronal pathway associated with the bladder by injecting PRV into the detrusor. An immunohistochemical method was used to stain Fos-protein encoded by the c-fos gene. Immunofluorescent staining for PRV was performed to evaluate changes in bladder-specific spinal neurons. RESULTS: Immunofluorescent staining with choline acetyltransferase (ChAT) revealed that the sacral parasympathetic nucleus (SPN) regions contained 9.8 PGNs/ section. In rats with chronic spinal cord injury by intravesical saline instillation, 82.4+/-10.3% of PGNs in SPN exhibited Fos-immunoreactive (IR). Two and a half days after PRV infection, PRV-IR PGNs were observed at 5.4 PGNs/ section, and 2.7+/-1.6% of them exhibited Fos-IR. Unlike ChAT-IR PGNs, PRV-IR PGNs are bladder-specific neurons and PRV-IR and Fos-IR cells found in the back of PRV-IR PGNs are bladder- specific interneurons. Three days after PRV infection, we observed many PRV-IR and Fos-IR cells in the dorsal commissure. These neurons are interneurons distributed in the bladder. CONCLUSION: We confirmed that in chronic spinal cord injury, the patterns of c-fos expression in bladder-specific spinal neurons were similar to those in voiding-reflex related spinal neurons, which had already been demonstrated earlier. We believe that our methodology can be applied to study interactions between voiding and other organs as well, such as the urethra and prostate.

Number, size and distribution of ganglion neurons in urinary bladder of rodents

Whole-mount preparations of urinary bladders stained with a modified Giemsa technique were obtained from three rodent species (Guinea-pig, Calomys callosus and the C57/BLJ isogenic mouse) to identify andestimate the relative number and size of ganglionic neurons within the wall of the organ. The istribution of the ganglia was not uniform among the three species: ganglia were concentrated around the ureteral orifices in the Guinea-pig, they were scattered throughout the organ in Calomys callosus, and they were oncentrated near the internal urethral orifice in the C57/BLJ mouse. In the Guinea-pig, the size of approximately 50 percent of the neurons lie in the range of 200 to 300 microns2. In Calomys callosus, 40 percent of the neurons lie in the range of 200 to 250 microns2, with 28 percnet in the range of 50 to 100 microns2. For the C57/BLJ mouse, approximately 60 percent of the neurons have an area of 250 to 400 microns2

Comparative study between cytology and dot ELISA for early detection of bladder cancer

A newly developed monoclonal antibody to keratinized Grade I squamous cell carcinoma was used in a dot enzyme-linked immunosorbent assay [ELISA] to test urine samples of 118 patients with bladder carcinoma, 291 patients with gentourinary pathology other than bladder carcinoma in addition to 550 healthy controls. The over all sensitivity of the dot ELISA was 90% among 118 patients with bladder carcinoma, 82% of the transitional cell carcinoma, 96% of the squamous cell carcinoma, 70% of the undifferentiated tumors and 100% of the adenocarcinoma were positive with this assay. The specificity was 90% in our sample population. A Comparative study of diagnosis by cytology and dot ELISA was carried out in 57 patients with bladder carcinoma. Dot ELISA, was found to be superior as a screening tool for high risk groups [P value <0.001 using the chi square test]. Cytolog detected 21% of transitional cell carcinoma, 68% of squamous cell carcinoma, 50% of adenocarinoma and 86% of undifferentiated tumors. The dot ELISA assay should be useful for screening high risk groups since it does not require sophisticated equipment, is non- invasive, does not require highly trained staff and can be performed in less than 30 minutes