Functional Expression of Transient Receptor Potential and Piezo1 Channels in Cultured Interstitial Cells of Human-Bladder Lamina Propria

The interstitial cells in bladder lamina propria (LP-ICs) are believed to be involved in sensing/afferent signaling in bladder mucosa. Transient receptor potential (TRP) cation channels act as mechano- or chemo-sensors and may underlie some of the sensing function of bladder LP-ICs. We aimed to investigate the molecular and functional expression of TRP channels implicated in bladder sensory function and Piezo1/Piezo2 channels in cultured LP-ICs of the human bladder. Bladder tissues were obtained from patients undergoing cystectomy. LP-ICs were isolated and cultured, and used for real-time reverse transcription-quantitative polymerase chain reaction, immunocytochemistry, and calcium-imaging experiments. At the mRNA level, TRPA1, TRPV2, and Piezo1 were expressed most abundantly. Immunocytochemical staining showed protein expression of TRPA1, TRPV1, TRPV2, TRPV4, TRPM8, as well as Piezo1 and Piezo2. Calcium imaging using channel agonists/antagonists provided evidence for functional expression of TRPA1, TRPV2, TRPV4, Piezo1, but not of TRPV1 or TRPM8. Activation of these channels with their agonist resulted in release of adenosine triphosphate (ATP) from LP-ICs. Inhibition of TRPV2, TRPV4 and Piezo1 blocked the stretch induced intracellular Ca2+ increase. Whereas inhibition of TRPA1 blocked H2O2 evoked response in LP-ICs. Our results suggest LP-ICs of the bladder can perceive stretch or chemical stimuli via activation of TRPV2, TRPV4, Piezo1 and TRPA1 channels. LP-ICs may work together with urothelial cells for perception and transduction of mechanical or chemical signals in human-bladder mucosa.

Intraoperative Problems and Solutions in Pneumovesicum Laparoscopic Cross-trigonal Ureteral Reimplantation in Children by a Beginner Surgeon

Objective: Many beginner surgeons feel anxious when first doing the procedure. Some may encounter many intraoperative difficulties or problems, resulting in abandoning the technique. We will demonstrate our methods and the solutions to major intraoperative problems.Materials and Methods: A beginner surgeon performed the operation on 13 children with VUR (20 ureters) who met the indications for surgery between October 2016 and August 2017. Age ranged from 2 to 7 years. Each operation comprised 2 main steps: anchoring the urinary bladder wall to the anterior abdominal wall under cystoscopic vision, followed by a cross-trigonal ureteral reimplantation under pneumovesicum laparoscopy. The intraoperative problems, postoperative care, and follow-up periods were recorded to identify surgical outcomes.Results: Most significant, intraoperative problems were air leakage, bleeding, tear of the bladder mucosa above the tunnel, and inability to insert a tube into the ureter pre- and post-reimplantation. Most problems could be managed. Only one case had to be converted to open reimplantation due to uncontrolled air leakage. Postoperatively, 2 patients had hydroureteronephrosis at 4 weeks, but it eventually spontaneously regressed. One patient had cystitis, treated with oral antibiotics. Between the 1-year and 4-year follow-up, no patients had hydroureteronephrosis or urinary tract infections (UTI).Conclusion: Pneumovesicum laparoscopic ureteral reimplantation is a feasible technique for beginner surgeons. Although many intraoperative problems may be encountered, most can be managed, resulting in the completion of the laparoscopic procedure.

Cystoscopic features of the ureteral orifices in children with vesicoureteral reflux

Purpose – to establish cystoscopic prognostic criteria for vesicoureteral reflux (VUR) in children. Materials and methods. Clinical material covers 270 patients with VUR II–IV grades aged 9 months to 14 years and 22 healthy children. The study included patients with VUR in the period of clinical and laboratory remission without symptoms of neurogenic bladder. During cystoscopy, the condition of the bladder mucosa was assessed; location, shape, hydrodistention degree, and ureteral orifices contractility. Results. Patients with VUR were diagnosed ureteral orifices in the form of: horseshoes – 127 (47.04%) patients, stadium – 106 (39.26%) and golf holes – 37 (13.7%). They were in the zones: A – 13 (4.81%) children, B – 154 (57.04%), C – 67 (24.81%), D – 36 (13.33%), and were characterized by the hydrodistention degree: H0 – 7 (2.59%) patients, H1 – 173 (64.07%), H2 – 60 (22.22%) and H3 – 30 (11.11%). In children with VUR, sluggish peristalsis of the ureter orifices clearly prevailed – 252 (93.33%) cases, relative to active peristalsis in only 18 (6.67%) patients. Conclusions. For ureteral orifices in the form of a stadium and with more pronounced signs of deepening, which are shifted to zone B and laterally to the sidewall of the bladder, with a hydrodistention degree above H1 has a positive association with VUR at the highest specificity of tests. Unfavorable prognostic diagnostic markers for effective minimally invasive interventions in patients with VUR should be considered ureteral orifices, which combine such morpho-topographic characteristics as pronounced signs of deepening to the shape of a golf hole, lateralization to the sidewall of the bladder in zone D, and hydrodistention H3 degree. The research was carried out in accordance with the principles of the Helsinki declaration. The study protocol was approved by the Local ethics committee of all participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interest was declared by the authors. Key words: vesicoureteral reflux, ureteral orifice, cystoscopy, children.

A rare cause of bladder outlet obstruction: Vesical diverticular calculus

Vesical diverticula are herniations of the bladder mucosa and submucosa through the muscular wall of the bladder. A calculus in diverticulum is an unusual finding. It is usually secondary to bladder outlet obstruction [1]. The patient typically presents with lower urinary tract symptoms, lower abdominal pain or abdominal distention. The present case is a 64-year-old male with a giant vesical diverticulum calculus. Cystolithotomy was done with primary closure of bladder, with repair of bladder diverticulum. Post-operatively, he recovered well with improved lower urinary tract function.

Effect of Electroacupuncture on Bladder Dysfunction via Regulation of MLC and MLCK Phosphorylation in a Rat Model of Type 2 Diabetes Mellitus

Previous studies observed have reported that electroacupuncture (EA) is effective in relieving diabetic bladder dysfunction (DBD); however, little is known about the mechanism. Therefore, we explored the effects and mechanisms of EA on DBD in streptozotocin–high-fat diet- (STZ–HFD-) induced diabetic rats. The Sprague-Dawley male rats were divided randomly into four groups: normal group, diabetes mellitus group (DM group), DM with EA treatment group (EA group), and DM with sham EA treatment group (sham EA group). After 8 weeks of EA treatment, the body weight, serum glucose, bladder weight, and cystometrogram were evaluated. The bladder wall thickness was examined by abdominal ultrasound imaging. After the transabdominal ultrasound measurements, hematoxylin-eosin (HE) staining was used to observe the bladder mucosa layer. The bladder detrusor smooth muscle cells (SMCs) and fibroblasts were observed under transmission electron microscopy (TEM). The phospho-myosin light chain (p-MLC), phospho-myosin light chain kinase (p-MLCK), and phospho-myosin phosphatase target subunit 1 (p-MYPT1) levels in the bladder were examined using Western blot. The bladder weight, serum glucose, bladder wall thickness, volume threshold for micturition, and postvoid residual (PVR) volume in the diabetic rats were significantly higher than those in the control animals. EA treatment significantly reduced the bladder weight, bladder wall thickness, volume threshold for micturition, and PVR volume in diabetic rats. EA caused a significant increase in the MLC dephosphorylation and MLCK phosphorylation levels in the group compared to the sham EA and model groups. EA reduced the infiltration of inflammatory cells in the bladder mucosa layer of diabetic rats. In addition, EA repaired the damaged bladder detrusor muscle of diabetic rats by reducing mitochondrial damage of the SMCs and fibroblasts. Therefore, EA could reduce the bladder hypertrophy to ameliorate DBD by reversing the impairment in the mucosa layer and detrusor SMCs, which might be mainly mediated by the regulation of p-MLC and p-MLCK levels.

Detection and Quantification of the E6 Oncogene in Bovine Papillomavirus Types 2 and 13 From Urinary Bladder Lesions of Cattle

Bovine papillomavirus types 2 and 13 can induce tumors in both the cutaneous and mucosal epithelia of cattle. These viral types are associated with the development of benign cutaneous papillomas and malignant lesions in the urinary bladders of cattle, with the latter being known as bovine enzootic hematuria. Among the viral oncoproteins encoded by Deltapapillomavirus DNA, the E6 oncoprotein has an important role in cell proliferation and might be related to cancer initiation and promotion. The aim of this study was to present a standardized SYBR Green-based quantitative PCR for detection and quantification of the bovine papillomavirus 2 and 13 E6 oncogenes in urinary bladder samples from cattle. Twenty-four urinary bladders from cattle displaying tumors (n = 12) and normal bladder mucosa (n = 12) were tested by quantitative PCR. Of the 12 urinary bladders with tumors, six presented bovine papillomavirus 2 DNA concentrations ranging from 1.05 × 104 to 9.53 × 103 copies/μL, while two had bovine papillomavirus 13 DNA amplified at concentrations of 1.30 × 104 to 1.23 × 104 copies/μL. The healthy bladder mucosa samples were negative for both bovine papillomaviruses. Once the results were confirmed by conventional PCR and direct sequencing, the quantitative PCR assay developed in this study was shown to be a sensitive and specific tool for detecting and quantifying the E6 ORF of bovine papillomavirus 2 and 13 in a variety of clinical samples. Our findings of identification of bovine papillomavirus 2 and 13 DNA in urothelial tumors from cattle suffering from bovine enzootic hematuria agree with data from previous studies, representing the first detection of bovine papillomavirus 13 DNA in malignant bladder lesions of cattle from Brazil.

Tadalafil ameliorates bladder overactivity by restoring insulin-activated detrusor relaxation via the bladder mucosal IRS/PI3K/AKT/eNOS pathway in fructose-fed rats

The pathophysiologies of metabolic syndrome (MS) and overactive bladder (OAB) might overlap. Using fructose-fed rats (FFRs) as a rodent model of MS we investigated the effects of tadalafil (a phosphodiesterase type 5 inhibitor) on the dysregulated insulin signalling in the bladder mucosa and bladder overactivity. Micturition behaviour was evaluated. Concentration–response curves on detrusor relaxation to insulin stimulation were examined. Expression and phosphorylation of proteins in the insulin signalling pathway were evaluated by Western blotting. Levels of detrusor cGMP and urinary nitrite and nitrate (NOx) were measured. We observed FFRs exhibited metabolic traits of MS, bladder overactivity, and impaired insulin-activated detrusor relaxation in organ bath study. A high-fructose diet also impeded insulin signalling, reflected by overexpression of IRS1/pIRS1Ser307 and pIRS2Ser731 and downregulation of PI3K/pPI3KTyr508, AKT/pAKTSer473, and eNOS/peNOSSer1177 in the bladder mucosa, alongside decreased urinary NOx and detrusor cGMP levels. Tadalafil treatment restored the reduced level of mucosal peNOS, urinary NOx, and detrusor cGMP, improved the insulin-activated detrusor relaxation, and ameliorated bladder overactivity in FFRs. These results suggest tadalafil may ameliorate MS-associated bladder overactivity by restoring insulin-activated detrusor relaxation via molecular mechanisms that are associated with preservation of IR/IRS/PI3K/AKT/eNOS pathway in the bladder mucosa and cGMP production in the bladder detrusor.

Epidemiological and Diagnostic Aspects of Bladder Bilharziomas in the Urology Department of Idrissa Pouye General Hospital (HOGIP)

Objective. The aim of our study is to assess the diagnostic aspects of bladder bilharzioma in the Urology Department of Idrissa Pouye General Hospital (Senegal). Materials and Methods. It is a descriptive study from January 2013 to December 2018. The patients included in the study were those who had anatomopathological examination of bladder biopsy that showed a schistosomiasis pseudotumor of the bladder. The variables studied were sociodemographic, clinical symptoms, imaging findings, histology, and treatment. The data have been saved and analyzed by the 2013 Excel software. Results. Thirteen patients were included in our study. The average age was 27 ± 12.1 years. Sex ratio was 1.6. The majority of the patients were from the northern part of Senegal. Hematuria was the main symptom for all the patients. Cystoscopy was performed for all the patients and showed 5 granulomas and 8 fibrocalcic polyps. A transurethral resection of the bladder was performed, and treatment with praziquantel (40 mg/kg of bodyweight) has been carried out. One patient presented precancerous lesions such as metaplasia and dysplasia of the bladder mucosa. After a median follow-up of 40 months (6–57 months), ten patients had a favorable clinical and endoscopic outcome. Conclusion. Granulomas and fibrocalcic polyps of the bladder mucosa are, respectively, confused with squamous cell carcinoma and bladder lithiasis in endemic areas of schistosomiasis. Good cystoscopy interpretation can provide the diagnosis of bladder bilharzioma and start the treatment.

Sheep urinary tract architecture is not affected by acute urethral obstruction

Urolithiasis affects the urinary tract of small ruminants, thereby requiring the animal to prematurely terminate breeding. Morphometric study of organs can be used as a diagnostic method. Thus, this study aimed to describe the macroscopic, histopathological, and histomorphometric changes in the urinary tract of sheep with urolithiasis. For this purpose, 14 healthy male Santa Inês sheep, approximately 90 days old, were studied and fed an experimental diet. After the development of urolithiasis, the animals were reorganized into two groups, D1 (without urolithiasis) and D2 (with urolithiasis) for comparative data analysis. Sheep were necropsied to evaluate the pathological changes, followed by macroscopic morphometric analysis, and the histopathological and histomorphometric characteristics were described. Urethral necrosis and a full urinary bladder were observed in all animals that developed the disease. The comparison between sheep with and without urolithiasis showed no significant difference (P < 0.05) in the evaluated macroscopic morphometric variables, except for the right ureter width. Regarding the histopathological evaluation, multifocal areas of mild to moderate congestion within the glomerular tufts and protein in the tubular lumen of the kidneys were observed. In the liver, mild to moderate fatty degeneration was noted in the centrolobular regions, and an ulcerated focal area in the bladder mucosa was observed in only one animal. The present study demonstrated that the formulated diet was effective in inducing clinical disease. In acute obstructive urolithiasis in sheep tissue, lesions in the liver and urinary tract were observed, although there were no significant histomorphometric changes.