Urothelial Injury Research Articles

Urinary antimicrobial peptides: Potential novel biomarkers of obstructive uropathy

Antimicrobial peptides (AMPs) have historically been evaluated for their role in protecting against uropathogens. However, there is mounting evidence to support their expression in noninfectious injury, with unclear meaning as to their function. It is possible that AMPs represent urothelial injury. Urinary tract obstruction is known to alter the urothelium; however, AMPs have not been evaluated for expression in this noninfectious injury. A pilot study to compare urinary AMP expression in children undergoing surgical intervention for ureteropelvic junction obstruction (UPJO) with nonobstructed controls. Bladder urine was collected from consenting/assenting pediatric patients with UPJO at intervention. Control bladder urines were obtained from age-matched and sex-matched healthy children without known obstruction or infection. Enzyme-linked immunosorbent assays were run for the following AMPs: β defense 1 (BD-1), neutrophil gelatinase-associated lipocalin (NGAL), cathelicidin (LL-37), hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein (HIP/PAP), and human α defensin 5 (HD-5); and normalized to urine creatinine. Results were analyzed with Student's t-test or Mann-Whitney U test, when appropriate, and receiver operating characteristic curves. A P-value of <0.05 was considered significant. Thirty bladder urine samples were obtained from children with UPJO at the time of decompressive intervention. Mean patient age was 4.7 years (range 0.3-18.4); 20 (67%) patients were male. Fifteen bladder urine samples were obtained from age-matched and sex-matched controls. Urinary AMP levels were significantly higher in UPJO patients than controls for BD-1 (P=0.015), NGAL (P<0.001), LL-37 (P<0.001), and HIP/PAP (P=0.046). Optimal threshold values of these AMPs were determined, with each demonstrating significant odds ratios of predicting urinary obstruction. Certain urinary AMPs are altered even in noninfectious urinary tract pathology. This represents a novel induction of AMP expression, as the current study is the first to report elevations in BD-1 and HIP/PAP in urinary tract obstruction. This suggests other roles for these AMPs outside of their antimicrobial properties, and likely is a reflection of the urothelial and tubular stress resulting from obstructive uropathy. Induction of AMPs BD-1, NGAL, LL-37, and HIP/PAP was found to occur in urinary tract obstruction. Further evaluation of AMP expression as a biomarker of uroepithelial injury outside of infection is indicated.

The anti-tumor effect of intravesical administration of normal urothelial cells on bladder cancer

Background aimsUrothelial bladder cancer (UBC) is the second most common cancer of the genitourinary tract and for advanced forms of the disease it has a high mortality rate. There are no approved new molecularly targeted agents or chemotherapeutics for advanced UBC beyond cisplatin-based chemotherapy except the recently approved anti-programmed death ligand 1 (anti-PD-1/PD-L1) antibody. With complex genetic and epigenetic alterations in tumors, despite several druggable targets identified, to cure UBC is still a challenging unmet medical need. Like other cancers, UBC to the host body is considered as a wound, aging stem cell disease and immunosuppressive disorder. Therefore, we proposed a novel cellular approach to target the host body by intravesical instilling of normal urothelial cells that could repair the injury and reduce inflammation by activating body-reparative capacity and because non-self cells are transplanted, host body immune responses could be induced in the tumor microenvironment of UBC to restrain and even eliminate tumor cells. MethodsIn this study, we isolated and expanded normal male murine urothelial cells and intravesically administered them into the bladders of female mice of two orthotopic bladder tumor models and one urothelial injury model. ResultsWe showed that the instillation of normal urothelial cells containing stem/progenitor cell population into bladders could have anti-tumor effect in orthotopic tumor models, possibly by activating immune responses and helping injured urothelium tissue recovery in a chemically induced urothelial injury model. ConclusionsOur findings could lead to an innovative and revolutionary cell therapy modality with normal urothelial cells as an effective and safe therapeutic option for UBC.

Carbenoxolone inhibits TRPV4 channel-initiated oxidative urothelial injury and ameliorates cyclophosphamide-induced bladder dysfunction.

Carbenoxolone (CBX) is a clinically prescribed drug for the treatment of digestive ulcer and inflammation. It is also a widely used pharmacological inhibitor of several channels in basic research. Given that the overactivity of several channels, including those inhibitable by CBX, underlies bladder dysfunction, we tested the potential therapeutic application and mechanism of CBX in the treatment of voiding dysfunction. In a mouse model of cystitis induced by cyclophosphamide (CYP), CBX administration prevented the CYP‐elicited increase in bladder weight, oedema, haemorrhage, and urothelial injury. CBX also greatly improved micturition pattern, as manifested by the apparently decreased micturition frequency and increased micturition volume. Western blot results showed that CBX suppressed CYP‐induced increase in protein carbonyls, COX‐2, and iNOS. Further analysis using cultured urothelial cells revealed that acrolein, the major metabolite of CYP, caused protein oxidation, p38 activation, and urothelial injury. These effects of acrolein were reproduced by TRPV4 agonists and significantly prevented by antioxidant NAC, p38 inhibitor SB203580, TRPV4 antagonist RN‐1734, and CBX. Further studies showed that CBX potently suppressed TRPV4 agonist‐initiated calcium influx and subsequent cell injury. CBX attenuated CYP‐induced cystitis in vivo and reduced acrolein‐induced cell injury in vitro, through mechanisms involving inhibition of TRPV4 channels and attenuation of the channel‐mediated oxidative stress. CBX might be a promising agent for the treatment of bladder dysfunction.

Urinary uroplakin expression in cyclophosphamide-induced rat cystitis model.

Cyclophosphamide (CYP) induces urothelial injury and causes excretion of cellular exudates at 24 h, followed by rapid restoration at 72 h. We investigated the role of urinary uroplakin II (UPII) levels in a CYP-induced cystitis model. For the purpose of this study, 10 controls and 26 CYP-injected female Sprague Dawley rats were killed at 24 h and 72 h postinjection. The vesical weight, severity of hematuria, and expression of UPII in the urinary bladder and urine were measured. CYP decreased the level of vesical UPII messenger RNA at 24 h, followed by rapid recovery at 72 h. Contrary to the negligible levels of urinary UPII and hematuria in controls, CYP treatment abruptly increased the excretion of urinary UPII at 24 h. The excretion had subsided at 72 h. Similarly, severe hematuria was observed at 24 h, with improvement at 72 h. However, some rats still exhibited hematuria at 72 h. CYP caused increase in vesical weight. The vesical weight at 24 h after CYP injection was negatively correlated with the vesical UPII level. Rats with significant hematuria demonstrated higher urinary UPII levels than those with insignificant hematuria. Vesical UPII could be an important barrier for early CYP-related injury, while the levels of urinary UPII may be associated with the severity of hematuria during dynamic periods in the urothelium.

Comparison of surgical intervention and non-surgery result in early prostate cancer

s / Urological Science 26 (2015) S50eS81 S57 The operation began with 7ecm midline incision above the umbilicus. A small hole of cystic wall was created and more than 6 L of yellowish clear fluid was drained. The hand-assisted device was placed in midline wound. While other two 10-mm trocar were placed in left lower abdomen and left anterior axillary line subcostally. The membrane of cyst was dissected as much as possible. Nephropexy was done to attach migrating left kidney to the back of the abdominal wall. The postoperative course was uneventful and the patient was discharged 3 days after the operation. Intravenous urography done 3 months later showed normal kidney position and bilateral patent ureters. Discussion: A huge renal cyst measuring more than 15 cm is an extremely rare. Cases presenting simply with progressive abdominal distention can lead to misdiagnosis, such as obesity or ascites. In our case, huge left renal cyst caused left kidney migration to right side, leading to initial misdiagnosis of right acute pyelonephritis. Surgery has been considered the mainstay of treatment for giant cyst. We performed hand assisted laparoscopic renal cyst unroofing and nephropexy with uneventful postoperative course and good outcome. NDP028: UNUSUAL URETHRAL FOREIGN BODIES Wei-Chung Hsiao , Sung-Lang Chen , Yu-Lin Kao , Wen-Jung Chen , Tzuo-Yi Hsieh , Shao-Chuan Wang . Division of Urology, Chung Shan Medical University Hospital, Taichung, Taiwan; 2 Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan Purpose: Urethral foreign bodies are uncommon clinical conditions. Most of the foreign bodies are self-introduced under the circumstances of autoerotic impulses, sexual curiosity, psychiatric illness, or rarely migration from adjacent organs. Retrieval of the urethral foreign body is casespecific due to wide variety of foreign bodies. Materials and Methods: A 75-year-old man was sent to emergency department after a motorcycle accident. Serial examinations including pelvic radiography and computed tomography discovered 5 fishhooks within the urethra unexpectedly. Ingrowth into urethral mucosa of the fishhooks with related inflammatory process was noted. Due to complexity of the fishhook position and pelvic fracture condition, cystourethroscopic retrieval of the foreign bodies was performed successfully in a two-step manner. Results: Self-inflicted urethral foreign bodies have a very low incidence. Major complications of urethral foreign bodies include trauma, infection, urine retention, and urethral stricture. Most of the foreign bodies of lower urinary tract can be treated with endourological modalities. The key point of urethral foreign body extraction relies on its physical attributes and morphology with the purpose of minimal urothelial injury. Conclusion: The urethral foreign bodies could be removed successfully by cystourethroscopic procedure without any complication, even in the presentation of mucosal ingrowth.

Diagnosing inflammation and infection in the urinary system via proteomics.

BackgroundCurrent methodology for the diagnosis of diseases in the urinary system includes patient symptomology, urine analysis and urine culture. Asymptomatic bacteriuria from urethral colonization or indwelling catheters, sample contamination from perineal or vaginal sources, and non-infectious inflammatory conditions can mimic UTIs, leading to uncertainty on medical treatment decisions.MethodsInnovative shotgun metaproteomic methods were used to analyze urine sediments from 120 patients also subjected to conventional urinalysis for various clinical reasons including suspected UTIs. The proteomic data were simultaneously searched for the presence of microbial agents, inflammation, immune responses against pathogens, and evidence of urothelial tissue injury. Hierarchical clustering analysis was performed to identify host protein patterns discerning UTI from urethral colonization and vaginal contamination of urine samples.ResultsOrganisms causing more than 98% of all UTIs and commensal microbes of the urogenital and perineal area were identified from 76 urine sediments with detection sensitivities estimated to be similar to urine culture. Proteomic data permitted a thorough evaluation of inflammatory and antimicrobial immune responses. Hierarchical clustering of the data revealed that high abundances of proteins from activated neutrophils were associated with pathogens in most cases, and correlated well with leukocyte esterase activities and leukocyte counts via microscopy. Proteomic data also allowed assessments of urothelial injury, by quantifying proteins highly expressed in red blood cells and contributing to the acute phase response. Lactobacillus and Gardnerella vaginalis were frequently identified suggesting urethral colonization and/or vaginal contamination of urine.ConclusionsA metaproteomic approach of interest for routine urine clinical diagnostics is presented. As compared to urinalysis and urine culture methods, the data are derived from a single experiment for a given sample and provide additional insights into presence or absence of inflammatory responses and vaginal contamination of urine specimens.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-015-0475-3) contains supplementary material, which is available to authorized users.

MP20-06 OPTICAL MONITORING OF DETRUSOR TISSUE OXYGEN SATURATION IN ACUTE LOWER URINARY TRACT INFECTION: A CASE CONTROL COMPARISON

You have accessJournal of UrologyInfections/Inflammation of the Genitourinary Tract: Kidney & Bladder II1 Apr 2015MP20-06 OPTICAL MONITORING OF DETRUSOR TISSUE OXYGEN SATURATION IN ACUTE LOWER URINARY TRACT INFECTION: A CASE CONTROL COMPARISON Babak Shadgan, Lynn Stothers, Andrew Macnab, Mark Nigro, and A Kajbafzadeh Babak ShadganBabak Shadgan More articles by this author , Lynn StothersLynn Stothers More articles by this author , Andrew MacnabAndrew Macnab More articles by this author , Mark NigroMark Nigro More articles by this author , and A KajbafzadehA Kajbafzadeh More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.978AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The diagnosis of lower urinary tract infection (LUTI) is based on history and clinical symptoms confirmed by positive urine culture. However, reliance on the clinical symptoms and history is problematic in certain clinical scenarios such as in young children and individuals with neurogenic bladder (e.g. spinal cord injury) where history and / or sensation of dysuria are lacking. The purpose of this study was to determine if a quantifiable measure of detrusor oxygen saturation, derived using a transcutaneous near-infrared spectroscopy (NIRS) system could distinguish between subjects with and without LUTI. METHODS Children referred to a pediatric urology clinic for evaluation of acute LUTI, and a matched control group, were studied. The presence or absence of UTI was confirmed by urine culture. Participants had transcutaneous measurement of an absolute measure of tissue oxygen saturation (TSI%) in their bladder wall, and a quadriceps muscle control site. The measurement was made using a miniature spatially-resolved wireless NIRS device (Portamon, Artinis Medical Technologies) applied over the bladder and on a quadriceps control site, with data collected at 10Hz for 1 minute with the patient in the supine position. NIRS wavelengths were 760 and 850 nanometers. To ensure that NIRS light penetrated bladder tissue the thickness of the supra-pubic fat layer was measured using ultrasound. Average measures of bladder wall TSI% (B.TSI%) and quadriceps TSI% (Q.TSI%) and their differences (TSI.diff) were calculated and compared between those with LUTI and controls by performing a two-way ANOVA. RESULTS Forty-eight patients met the inclusion criteria (24 LUTI and 24 controls). Comparing LUTI to controls B.TSI% and TSI.diff values were significantly higher in the LUTI group (p<0.0001), while Q.TSI% values were not significantly different. In all UTI patients the SR NIRS-derived TSI.diff value was equal to or higher than 5.2, while in all non-UTI subjects the TSI.diff was lower than 2.6. Table – Group comparison CONCLUSIONS NIRS optical monitoring of an absolute measure of bladder wall oxygenation may offer a means of screening for LUTI where history and/or clinical signs are not available or adequate. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e225 Peer Review Report Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Babak Shadgan More articles by this author Lynn Stothers More articles by this author Andrew Macnab More articles by this author Mark Nigro More articles by this author A Kajbafzadeh More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP20-05 BLADDER INSTILLATION THEARPY OF MESENCYMAL STEM CELL AND THE ADDITION OF MANNOSE IN A RAT CYSTITIS MODEL- MULTIPLE SUPPRESSIVE EFFECTS FOR EXCESS CYTOKINES

You have accessJournal of UrologyInfections/Inflammation of the Genitourinary Tract: Kidney & Bladder II1 Apr 2015MP20-05 BLADDER INSTILLATION THEARPY OF MESENCYMAL STEM CELL AND THE ADDITION OF MANNOSE IN A RAT CYSTITIS MODEL- MULTIPLE SUPPRESSIVE EFFECTS FOR EXCESS CYTOKINES Tokunori Yamamoto, Yasuto Funahashi, Majima Tsuyoshi, Takai Syun, Yoshihisa Matsukawa, Hideki Mizuno, and Momokazu Gotoh Tokunori YamamotoTokunori Yamamoto More articles by this author , Yasuto FunahashiYasuto Funahashi More articles by this author , Majima TsuyoshiMajima Tsuyoshi More articles by this author , Takai SyunTakai Syun More articles by this author , Yoshihisa MatsukawaYoshihisa Matsukawa More articles by this author , Hideki MizunoHideki Mizuno More articles by this author , and Momokazu GotohMomokazu Gotoh More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.977AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Mesencymal stem cell(MSC) is used as a clinical regeneration therapeutic agent for various organs. We examined the healing effects of intravesical MSC on damaged urothelium in a rat model of chemically induced cystitis. METHODS Rat(r) and human(h)-MSC instillation in the bladder of female Sprague Dawley® and nude rats to cyclophosphamide induce cystitis respectively. After 8 hour r-MSC(n=6) ,r and h MSC with mannose(MN) (n=6),vehicle(n=6) were administered in the bladder for 48 hour. Histopathology, urothelial permeability, cystometrogram and nociceptive behaviors were evaluated on day 2. Bladder inflammation were evaluated the cytokaines of bladder urine and tissue measured by multipulex system and urine myeloperoxidase(MPO) measured by ELISA. RESULTS MN increased cell activity of MSC in vitro. Bladder histological evaluation revealed polymorphological inflammatory cell infiltration and increase in inflammatory and protect of damaged urothelium(Fig) and cytokines in urine and the tissue and MPO in urine. h-MSC and h-MSC with MN were homing in interstitialis of bladder of nude rats(Fig). Evans blue over absorption in the bladder wall were decreased in MSC and MSC with MN treated rats. These findings, which were associated with urothelial injury and increased permeability. Cystometrogram demonstrated that the intercontraction interval were shorter in MSC and MSC with MN treated rats furhthermore. CONCLUSIONS MSC and MSC with MN accelerated via multiple cytokines suppressive effects for excess cytokines the repair of damaged urothelium, protected urothelial barrier function and suppressed bladder overactivity and nociception respectively. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e224-e225 Peer Review Report Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tokunori Yamamoto More articles by this author Yasuto Funahashi More articles by this author Majima Tsuyoshi More articles by this author Takai Syun More articles by this author Yoshihisa Matsukawa More articles by this author Hideki Mizuno More articles by this author Momokazu Gotoh More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Multiple impacted urethral metallic needles and screws (foreign bodies) associated with polyembolokoilamania.

This study aims to present the challenges faced in the management of multiple impacted foreign bodies, needles, and screws from the penile and bulbar urethra. A young man presented with complaint of a hard perineal swelling and passage of metallic nails per urethra. Pelvic radiograph revealed multiple foreign bodies (nails) in the penile and bulbar urethra. Successful cystoscopic removal of 11 foreign bodies comprising four large metallic screws and seven nail-like large sewing needles was done in two sessions. The most prevalent motivation for self-insertion of urethral foreign bodies is autoerotism/psychological impairment. Appropriate surgical technique guided by physical examination/ imaging with endoscopic removal is often successful, depending on the object's physical attributes and morphology while minimizing urothelial trauma and preserving voiding and erectile function. Follow-up cystourethroscopy is important for diagnosing any complications and urothelial injuries.

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Background: Homeostatic maintenance and repair of the bladder urothelium has been attributed to proliferation of keratin 5‐expressing basal cells (K5‐BC) with subsequent differentiation into superficial cells. Recent evidence, however, suggests that the intermediate cell layer harbors a population of progenitor cells. We use label‐retaining cell (LRC) methodology in conjunction with a clinically relevant model of uropathogenic Escherichia coli (UPEC)‐induced injury to characterize urothelial ontogeny during development and in response to diffuse urothelial injury. Results: In the developing urothelium, proliferating cells were dispersed throughout the K5‐BC and intermediate cells layers, becoming progressively concentrated in the K5‐BC layer with age. When 5‐bromo‐2‐deoxyuridine (BrdU) was administered during urothelial development, LRCs in the adult were found within the K5‐BC, intermediate, and superficial cell layers, the location dependent upon time of labeling. UPEC inoculation resulted in loss of the superficial cell layer followed by robust proliferation of K5‐BCs and intermediate cells. LRCs within the K5‐BC and intermediate cell layers proliferated in response to injury. Conclusions: Urothelial development and regeneration following injury relies on proliferation of K5‐BC and intermediate cells. The existence and proliferation of LRCs within both the K5‐BC and intermediate cell layers suggests the presence of two populations of urothelial progenitor cells. Developmental Dynamics 243::988–998, 2014. © 2014 Wiley Periodicals, Inc.

Intravesical Application of Rebamipide Promotes Urothelial Healing in a Rat Cystitis Model

Rebamipide is used as a topical therapeutic agent for various organs. We examined the healing effects of intravesical rebamipide on damaged urothelium in a rat model of chemically induced cystitis. Hydrochloride was injected in the bladder of female Sprague Dawley® rats to induce cystitis. On days 1 and 4 rebamipide (1 or 10mM) or vehicle was administered in the bladder and maintained for 1 hour. Histopathology, urothelial permeability, cystometrogram and nociceptive behaviors were evaluated on day 7. Also, tissue rebamipide concentrations after the 1-hour bladder instillation were quantified using high performance liquid chromatography. Intravesically administered rebamipide permeated the bladder, particularly in hydrochloride treated rats, and the pharmacologically effective tissue dose remained for greater than 6 hours. Bladder histological evaluation revealed polymorphological inflammatory cell infiltration and decreased positive staining for uroplakin 3A in hydrochloride treated rats. Scanning electron microscopy showed damaged tight junctions in the hydrochloride group. Evans blue absorption in the bladder wall was increased in hydrochloride treated rats. These findings, which were associated with urothelial injury and increased permeability, were dependently suppressed by the rebamipide treatment dose. Cystometrogram demonstrated that the intercontraction interval was shorter in hydrochloride treated rats but prolonged by rebamipide. The increased nociceptive behaviors observed after intravesical resiniferatoxin administration werealso suppressed by rebamipide. Intravesical rebamipide accelerated the repair of damaged urothelium, protected urothelial barrier function and suppressed bladder overactivity and nociception.

A population of progenitor cells in the basal and intermediate layers of the murine bladder urothelium contributes to urothelial development and regeneration.

Homeostatic maintenance and repair of the bladder urothelium has been attributed to proliferation of keratin 5-expressing basal cells (K5-BC) with subsequent differentiation into superficial cells. Recent evidence, however, suggests that the intermediate cell layer harbors a population of progenitor cells. We use label-retaining cell (LRC) methodology in conjunction with a clinically relevant model of uropathogenic Escherichia coli (UPEC)-induced injury to characterize urothelial ontogeny during development and in response to diffuse urothelial injury. In the developing urothelium, proliferating cells were dispersed throughout the K5-BC and intermediate cells layers, becoming progressively concentrated in the K5-BC layer with age. When 5-bromo-2-deoxyuridine (BrdU) was administered during urothelial development, LRCs in the adult were found within the K5-BC, intermediate, and superficial cell layers, the location dependent upon time of labeling. UPEC inoculation resulted in loss of the superficial cell layer followed by robust proliferation of K5-BCs and intermediate cells. LRCs within the K5-BC and intermediate cell layers proliferated in response to injury. Urothelial development and regeneration following injury relies on proliferation of K5-BC and intermediate cells. The existence and proliferation of LRCs within both the K5-BC and intermediate cell layers suggests the presence of two populations of urothelial progenitor cells.

Nephrogenic adenoma of the urethra presenting as hematuria.

Nephrogenic adenoma (NA) is a form of nephrogenic metaplasia with formation of benign epithelial tumours. NA arise from the urothelium as a result of reaction to urothelial injury like trauma, indwelling catheters, previous surgery, renal transplants or calculi. The exact aetiology of this lesion however still remains uncertain [1]. In view of its propensity to cause hematuria, NA needs to be considered and treated especially in the relevant background of previous surgical trauma in the urinary tract. We highlight the presentation and management of this lesion in the following case report.

Protamine Sulfate Induced Bladder Injury Protects from Distention Induced Bladder Pain

Bladder pain is a debilitating symptom of many urological conditions. There is no generally effective treatment. Abnormal urothelial turnover is common to multiple disease states but the specific components of urothelial injury and the resulting molecular signals that lead to bladder pain are unknown. We examined mouse models of bladder injury induced by uropathogenic Escherichia coli, protamine sulfate (Sigma®) and bacterial lipopolysaccharide to identify cellular and molecular correlates underlying pain sensitization in response to the stimuli. C57BL/6 female mice (Jackson Laboratory, Bar Harbor, Maine) were given intravesicular protamine sulfate, lipopolysaccharide or uropathogenic E. coli. The impact of each on nociception was determined by measuring the evoked visceromotor response to bladder distention 24 hours after inoculation. Levels of pyuria and tissue inflammation were examined by urinary cytology and tissue histology. Quantitative polymerase chain reaction and gene expression analysis were used to identify injury profiles associated with nociception. Protamine sulfate treatment was significantly analgesic upon bladder distention. Protamine treated bladders did not show pyuria or extensive tissue damage. Protamine injury was associated with a global decrease in the expression of inflammation associated genes. In contrast, uropathogenic E. coli injury significantly increased the nociceptive response to bladder distention. Lipopolysaccharide treatment did not affect nociception. Finally, injury induced expression of inflammation associated genes correlated with nociceptive responses. Protamine treatment of the bladder is analgesic and tissue protective, and it suppresses the inflammatory cytokine expression normally associated with nociception. Also, the injury modalities that result in differential tissue response patterns provide an innovative method for identifying mediators of visceral pain.

Hyperplasia as a mechanism for rapid resealing urothelial injuries and maintaining high transepithelial resistance

When the urothelial barrier, i.e., the blood-urine barrier, is injured, rapid resealing of the injury is crucial for the normal functioning of the organism. In order to investigate the mechanisms required for rapid resealing of the barrier, we established in vitro models of hyperplastic and normoplastic urothelia. We found that hyperplastic urothelia achieve significantly higher transepithelial resistance (TER) than normoplastic urothelia. However, the expression of cell junctional (claudin-8, occludin, E-cadherin) and differentiation-related proteins (cytokeratin 20 and uroplakins) is weaker in hyperplastic urothelia. Further investigation of cell differentiation status at the ultrastructural level confirmed that superficial urothelial cells (UCs) in hyperplastic urothelial models achieve a lower differentiation stage than superficial UCs in normoplastic urothelial models. With the establishment of such in vitro models and the aid of TER measurements, flow cytometry, molecular and ultrastructural analysis, we here provide unequivocal evidence that the specific cell-cycle distribution and, consequently, the number of cell layers have a significant influence on the barrier function of urothelia. We demonstrate the importance of hyperplasia for the rapid restoration of the urothelial barrier and the maintenance of high TER until the UCs reach a highly differentiated stage and restoration of the urothelial barrier after injury is complete. The information that this approach provides is unique and we expect that further exploitation of hyperplastic and normoplastic urothelial models in future studies may advance our understanding of blood-urine barrier development and functionality.

Complications following 573 Percutaneous Renal Radiofrequency and Cryoablation Procedures

Purpose To review complications related to percutaneous renal tumor ablation. Materials and Methods Prospectively collected data related to renal radiofrequency (RF) ablation and cryoablation procedures performed from May 2000 through November 2010 were reviewed. This included 573 renal ablation procedures performed in 533 patients to treat 633 tumors. A total of 254 RF ablation and 311 cryoablation procedures were performed; eight patients underwent simultaneous RF ablation and cryoablation. The mean age of patients at the time of the procedure was 70 years (range, 24–93 y), and 382 of 573 procedures (67%) were performed in male patients. Complications were recorded according to the Clavien–Dindo classification scheme. Duration of hospitalization was also documented. Results Of the 573 procedures, 63 produced complications (11.0% overall complication rate). There were 66 reported complications, of which 38 (6.6% of total procedures) were Clavien–Dindo grade II–IV major complications; there were no deaths. Major complication rates did not differ statistically ( P = .15) between cryoablation (7.7%; 24 of 311) and RF ablation (4.7%; 12 of 254). Of the complications related to cryoablation, bleeding and hematuria were most common. Bleeding during cryoablation was associated with advanced age, increased tumor size, increased number of cryoprobes, and central position ( P < .05). Of those treated with RF ablation, nerve and urothelial injury were most common. Mean hospitalization duration was 1 day for RF ablation and cryoablation. Conclusions Complications related to percutaneous renal ablation are infrequent. Recognition of potential complications and associated risk factors can allow optimization of periprocedural care.

Uropathogenic E. coli Promote a Paracellular Urothelial Barrier Defect Characterized by Altered Tight Junction Integrity, Epithelial Cell Sloughing and Cytokine Release

The urinary bladder is a common site of bacterial infection with a majority of cases attributed to uropathogenic Escherichia coli. Sequelae of urinary tract infections (UTIs) include the loss of urothelial barrier function and subsequentclinical morbidity secondary to the permeation of urine potassium, urea and ammonia into the subepithelium.To date there has been limited research describing the mechanism by which this urothelial permeabilitydefect develops. The present study models acute uropathogenic E. coli infection invitro using intact canine bladder mucosa mounted in Ussing chambers to determine whether infection induces primarily a transcellular or paracellular permeability defect. The Ussing chamber sustains tissue viability while physically separatingsubmucosal and lumen influences, so this model is ideal for quantitative measurement of transepithelial electrical resistance (TER) to assess alterations of urothelial barrier function. Using this model, changes in both tissue ultrastructure and TER indicated that uropathogenic E. coli infection promotes a paracellular permeability defect associated with the failure of umbrella cell tight junction formation and umbrella cell sloughing. In addition, bacterial interaction with the urothelium promoted secretion of cytokines from the urinarybladder with bioactivity capable of modulating epithelial barrier function including tumour necrosis factor-α, interleukin (IL)-6 and IL-15. IL-15 secretion by the infected bladder mucosa is a novel finding and, because IL-15 plays key roles in reconstitution of tight junction function in damaged intestine, this study points to a potential role for IL-15 in UTI-induced urothelial injury.

Nephrogenic Adenoma of the Bladder in a Prune Belly Syndrome Patient: Case Report and Review of the Literature

Nephrogenic adenoma (NA) is a rare lesion of the urinary tract widely considered to be a metaplastic response to urothelial injury. Herein, we present the case of an 8-year-old male with prune belly syndrome who presented with gross hematuria. Investigation revealed a bladder mass; however, upon cystoscopic examination, multiple polypoid lesions were identified. Microscopic examination revealed NA of the bladder. To our knowledge, this is the second reported case of NA of the bladder in association with prune belly syndrome.

An EGFR-ERK-SOX9 Signaling Cascade Links Urothelial Development and Regeneration to Cancer

Like many carcinomas, urothelial carcinoma (UroCa) is associated with chronic injury. A better understanding of this association could inform improved strategies for preventing and treating this disease. We investigated the expression, regulation, and function of the transcriptional regulator SRY-related high-mobility group box 9 (Sox9) in urothelial development, injury repair, and cancer. In mouse bladders, Sox9 levels were high during periods of prenatal urothelial development and diminished with maturation after birth. In adult urothelial cells, Sox9 was quiescent but was rapidly induced by a variety of injuries, including exposure to the carcinogen cyclophosphamide, culture with hydrogen peroxide, and osmotic stress. Activation of extracellular signal-regulated kinases 1/2 (ERK1/2) was required for Sox9 induction in urothelial injury and resulted from activation of the epidermal growth factor receptor (Egfr) by several Egfr ligands that were dramatically induced by injury. In UroCa cell lines, SOX9 expression was constitutively upregulated and could be suppressed by EGFR or ERK1/2 blockade. Gene knockdown showed a role for SOX9 in cell migration and invasion. Accordingly, SOX9 protein levels were preferentially induced in invasive human UroCa tissue samples (n = 84) compared with noninvasive cancers (n = 56) or benign adjacent urothelium (n = 49). These results identify a novel, potentially oncogenic signaling axis linking urothelial injury to UroCa. Inhibiting this axis is feasible through a variety of pharmacologic approaches and may have clinical utility.

Irreversible electroporation (IRE): a novel method for renal tissue ablation

• To evaluate the effects of irreversible electroporation (IRE) on renal parenchyma and the renal collecting system in a porcine model. • Eight female Yorkshire pigs underwent a series of laparoscopic ablations using either monopolar or bipolar IRE (Angiodynamics, Queensbury, NY, USA). • The pigs were killed between 10 min and 14 days after IRE, and the kidneys were harvested for gross and histological analysis, including NADH staining for cellular viability. • In all, 24 ablations were performed and all the pigs survived without complications. • Initial gross lesions were diffusely haemorrhagic, decreasing progressively in size (30-40%) to small white scars over the 14-day period. • Immediately after IRE, ablated tissue was characterized by diffuse tubular desquamation, eosinophilia, and nuclear pyknosis, with absence of cellular viability by NADH. • At 7 days after IRE, there was diffuse cellular necrosis with early peripheral granulation changes, and by 14 days there was marked tissue granulation, chronic inflammation, and dystrophic calcification with early fibrosis and cellular contraction. • Initial patchy urothelial injury and ulceration showed signs of repair and viability by 14 days after IRE. • Renal IRE in the porcine kidney leads to predictable histological changes characteristic of cellular death within 1 h of ablation, with relative urothelial sparing. • Further animal studies are warranted to determine safety and efficacy of this novel ablation technology.