Bladder Weight Research Articles

PD01-02 GENE THERAPY WITH REPLICATION-DEFICIENT HERPES SIMPLEX VIRUS (HSV) VECTORS ENCODING PORELESS TRPV1 (PL) OR PROTEIN PHOSPHATASE 1α (PP1α) IN A RAT MODEL OF HYDROGEN PEROXIDE-INDUCED CYSTITIS

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Interstitial Cystitis I1 Apr 2017PD01-02 GENE THERAPY WITH REPLICATION-DEFICIENT HERPES SIMPLEX VIRUS (HSV) VECTORS ENCODING PORELESS TRPV1 (PL) OR PROTEIN PHOSPHATASE 1α (PP1α) IN A RAT MODEL OF HYDROGEN PEROXIDE-INDUCED CYSTITIS Shun Takai, Tsuyoshi Majima, Takahiro Shimizu, Naoki Wada, Nobutaka Shimizu, Takahisa Suzuki, Eiichiro Takaoka, Momokazu Gotoh, William Goins, Joseph Glorioso, and Naoki Yoshimura Shun TakaiShun Takai More articles by this author , Tsuyoshi MajimaTsuyoshi Majima More articles by this author , Takahiro ShimizuTakahiro Shimizu More articles by this author , Naoki WadaNaoki Wada More articles by this author , Nobutaka ShimizuNobutaka Shimizu More articles by this author , Takahisa SuzukiTakahisa Suzuki More articles by this author , Eiichiro TakaokaEiichiro Takaoka More articles by this author , Momokazu GotohMomokazu Gotoh More articles by this author , William GoinsWilliam Goins More articles by this author , Joseph GloriosoJoseph Glorioso More articles by this author , and Naoki YoshimuraNaoki Yoshimura More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.182AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Increased afferent excitability is considered to be an important pathophysiological basis of interstitial cystitis/bladder pain syndrome (IC/BPS) and overactive bladder (OAB). Also, transient receptor potential vanilloid 1 (TRPV1) receptors are known to be involved in afferent sensitization. Therefore, we investigated the effect of gene therapy with HSV vectors encoding PL or PP1α, a negative regulator of TRPV1, using a rat model of long-lasting cystitis induced by hydrogen peroxide (HP). METHODS HSV vectors encoding green fluorescent protein (GFP), PL or PP1α were injected into the bladder wall of female SD rats. One week later, 1% HP or normal saline (NS) was administered into the bladder. Awake cystometry (CMG), resiniferatoxin (RTX)-induced nociceptive behaviors such as licking and freezing, NGF mRNA expression in the bladder, bladder weight and histology were evaluated 2 weeks after viral injection. RESULTS In CMG, the GFP/HP group showed a significant decrease in intercontraction intervals compared to the GFP/NS group, which were significantly prolonged by 57.8% and 68.0% in PL/HP and PP1α/HP groups (p<0.01), respectively. The number of freezing behavior was significantly lower in PL/HP and PP1α/HP groups by 86.1% and 93.5%, respectively, compared to the GFP/HP group. Compared with the GFP/NS group, the GFP/HP group had significantly heavier bladder weight, whereas the PL/HP group and PP1α/HP group showed significantly lighter bladder weight than the GFP/HP group. Hematoxylin and eosin staining of bladder sections showed substantial inflammation characterized by inflammatory cell infiltration, and detrusor hypertrophy in the bladder in the GFP/HP group compared with GFP/NS group, which were alleviated in PL/HP and PP1α/HP groups. In RT-PCR, the GFP/HP group showed higher significantly (p<0.05) expression of NGF mRNA in the bladder mucosa than the GFP/NS group, which was significantly decreased in the PL/HP and PP1α/HP groups (p < 0.05). CONCLUSIONS HSV vectors-mediated gene delivery of PL or PP1α significantly reduced bladder overactivity and enhanced bladder pain sensitivity in HP cystitis rats. Also, both treatments can reduce bladder inflammatory changes and increased bladder weight at least in part through amelioration of NGF overexpression in the bladder mucosa. Thus, HSV-mediated TRPV1-targeting gene therapy could be effective for the treatment of IC/BPS including Hunner-type IC that is often associated with bladder inflammation. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e46 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Shun Takai More articles by this author Tsuyoshi Majima More articles by this author Takahiro Shimizu More articles by this author Naoki Wada More articles by this author Nobutaka Shimizu More articles by this author Takahisa Suzuki More articles by this author Eiichiro Takaoka More articles by this author Momokazu Gotoh More articles by this author William Goins More articles by this author Joseph Glorioso More articles by this author Naoki Yoshimura More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP85-03 THE THERAPEUTIC EFFECT OF TRPV4 ACTIVATION IN THE BLADDER ON UNDERACTIVE BLADDER IN RATS WITH PELVIC NERVE CRUSH

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Neurogenic Voiding Dysfunction I1 Apr 2017MP85-03 THE THERAPEUTIC EFFECT OF TRPV4 ACTIVATION IN THE BLADDER ON UNDERACTIVE BLADDER IN RATS WITH PELVIC NERVE CRUSH Ei-ichiro Takaoka, Shun Takai, Takahisa Suzuki, Nobutaka Shimizu, Joonbeom Kwon, Hiroki Okada, Hiroyuki Nishiyama, Naoki Yoshimura, and Christopher J. Chermansky Ei-ichiro TakaokaEi-ichiro Takaoka More articles by this author , Shun TakaiShun Takai More articles by this author , Takahisa SuzukiTakahisa Suzuki More articles by this author , Nobutaka ShimizuNobutaka Shimizu More articles by this author , Joonbeom KwonJoonbeom Kwon More articles by this author , Hiroki OkadaHiroki Okada More articles by this author , Hiroyuki NishiyamaHiroyuki Nishiyama More articles by this author , Naoki YoshimuraNaoki Yoshimura More articles by this author , and Christopher J. ChermanskyChristopher J. Chermansky More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2665AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Incomplete bladder emptying due to detrusor underactivity (DU) is a significant urological problem underlying underactive bladder (UAB). Also, TRPV4 has been reported to be one of the mechanosensitive channels expressed in the bladder. In this study, we sought to produce a consistent rat model of UAB with the modification of our previous model of pelvic nerve crush (PNC) (2016 AUA) and evaluated the therapeutic effect of intravesical application of a TRPV4 agonist on the UAB condition. METHODS In female Sprague-Dawley rats, the visceral branches of bilateral pelvic nerves were identified near the internal iliac vessels and bilateral PNC was made by two times of nerve compression of either side with each 20-seconds duration using sharp forceps. After 10 days, awake cystometrograms (CMG) were recorded in sham control and PNC rats. Then, in both groups, a TRPV4 agonist (GSK1016790A) was continuously administered into the bladder and the CMG parameters were compared before and after intravesical drug administration. RESULTS The bladder weight was significantly increased in PNC rats vs. control rats. In CMG, PNC rats showed significant increases in voided volume, post-void residual urine volume, and residual urine rate compared to control rats. PNC rats also revealed the significant increases in intercontraction intervals (ICI), a number of non-voiding contractions, and threshold pressure while the amplitude during voiding was significantly decreased (table). In the TRPV4 administration study, intravesical application of 1.5µM of GSK 1016790A significantly decreased ICI, voided volume, and post-void residual urine volume in PNC rats while it did not significantly affect any CMG parameters in control rats. CONCLUSIONS Rats with pelvic nerve injury induced by the modified PNC method, which showed the characteristics of DU, seem to be an appropriate model for evaluation of peripheral neurogenic mechanisms of UAB. Also TRPV4 that reduced the bladder capacity and residual urine volume could be a potential target for the treatment of UAB. © 2017FiguresReferencesRelatedDetailsCited byKaplan S (2017) Re: Qualitative Exploration of the Patient Experience of Underactive BladderJournal of Urology, VOL. 199, NO. 3, (588-589), Online publication date: 1-Mar-2018. Volume 197Issue 4SApril 2017Page: e1148-e1149 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Ei-ichiro Takaoka More articles by this author Shun Takai More articles by this author Takahisa Suzuki More articles by this author Nobutaka Shimizu More articles by this author Joonbeom Kwon More articles by this author Hiroki Okada More articles by this author Hiroyuki Nishiyama More articles by this author Naoki Yoshimura More articles by this author Christopher J. Chermansky More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP94-16 OXIDATIVE STRESS-RELATED ALTERATIONS IN THE BLADDER OF A SHORT-PERIOD DIABETES TYPE-2 RAT MODEL

You have accessJournal of UrologyBladder & Urethra: Anatomy, Physiology & Pharmacology II1 Apr 2017MP94-16 OXIDATIVE STRESS-RELATED ALTERATIONS IN THE BLADDER OF A SHORT-PERIOD DIABETES TYPE-2 RAT MODEL Panagiota Tsounapi, Masashi Honda, Fotios Dimitriadis, Yusuke Kimura, Shogo Shimizu, Bunya Kawamoto, Katsuya Hikita, Motoaki Saito, Nikolaos Sofikitis, and Atsushi Takenaka Panagiota TsounapiPanagiota Tsounapi More articles by this author , Masashi HondaMasashi Honda More articles by this author , Fotios DimitriadisFotios Dimitriadis More articles by this author , Yusuke KimuraYusuke Kimura More articles by this author , Shogo ShimizuShogo Shimizu More articles by this author , Bunya KawamotoBunya Kawamoto More articles by this author , Katsuya HikitaKatsuya Hikita More articles by this author , Motoaki SaitoMotoaki Saito More articles by this author , Nikolaos SofikitisNikolaos Sofikitis More articles by this author , and Atsushi TakenakaAtsushi Takenaka More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2919AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Diabetes type-2 accounts for almost 90% of diabetes cases worldwide. Diabetes among other complications induces bladder dysfunction. In the current study we aimed to create a short-period diabetes type-2 model in order to investigate oxidative stress-related alterations in the bladder in the initiation of the disease. Additionally antioxidant treatment with resveratrol or taurine was provided in order to examine whether it is possible to prevent these alterations in the very beginning of the disease. METHODS Diabetes was induced in 8-week-old male Wistar rats with a single dose of streptozotocin (40mg/kg) intraperitoneally (i.p.). The next day they were randomly separated into 3 groups and 14 days feeding with a high fat diet followed. One group received no treatment (DM group), another group received orally resveratrol (10mg/Kg; Resv group) and the third one received taurine i.p. (1g/Kg; Tau group). Age matched control animals were used and were fed with normal diet (Control group). Two weeks later animals were sacrificed and bladder were processed for histological evaluation, measurements of malondialdehyde (MDA) and immunohistochemistry (IHC) for oxidative stress markers. RESULTS At the end of 2 weeks all diabetic groups had significantly lower body weight compared to the Control group. DM group demonstrated significantly higher ratio of bladder weight to body weight compared to the Control (Figure 1). Histological evaluation demonstrated mild damage of the bladder tissue in the DM group such as abruption of the mucosa from the muscularis as well as edema in the transitional epithelium. All these alterations were not observed in the treatment groups. MDA levels in the bladder were significantly larger in the DM group compared with Control, Resv or Tau group (Figure 1). Fourteen days of diabetes without treatment in the DM group induced moderate to strong intensity and positivity for oxidative stress marker MDA, 4-Hydroxynonenal and DNA oxidative stress marker 8-deoxyguanosine compared with the other three groups. CONCLUSIONS The prompt diagnosis of diabetes can be crucial for the progression of the disease. Specifically in the bladder, it appears that both mild damage in structural level as well as oxidative damage in molecular level can be prevented by resveratrol or taurine treatment. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1252 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Panagiota Tsounapi More articles by this author Masashi Honda More articles by this author Fotios Dimitriadis More articles by this author Yusuke Kimura More articles by this author Shogo Shimizu More articles by this author Bunya Kawamoto More articles by this author Katsuya Hikita More articles by this author Motoaki Saito More articles by this author Nikolaos Sofikitis More articles by this author Atsushi Takenaka More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

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.

A Chinese Medicine Formula “Xian‐Jia‐Tang” for Treating Bladder Outlet Obstruction by Improving Urodynamics and Inhibiting Oxidative Stress through Potassium Channels

The aim of this study is to investigate efficacy of a traditional Chinese medicine formula (named Xian-Jia-Tang, XJT) on bladder outlet obstruction (BOO) in rats and explore its mechanisms. Total 80 BOO model rats were established and randomly divided into 4 groups: physiological saline, XJT, Cesium Chloride (CC), and XJT and CC groups. Meanwhile, 12 rats were used as normal control. Bladder weight and urodynamics were measured. Oxidative stress level and mRNA expressions of potassium channels gene were detected in detrusor. The mRNA and protein levels of hypoxia inducible factor-α (HIF-α) in detrusor were detected by RT-PCR and Western blot. BOO model rats showed significantly higher bladder weight and abnormal urodynamics. XJT significantly improved the abnormal urodynamics and inhibited the oxidative stress and changes of mRNA levels of potassium channels genes in detrusor of BOO model rats. Moreover, KATP and SK2/3 mRNA were overexpressed in BOO model rats treated by XJT. Besides, the significantly increased levels of HIF-α mRNA and protein were also inhibited by XJT. However, these inhibition effects of XJT were weakened by CC. XJT could effectively improve the urodynamics and inhibit the oxidative stress caused by hypoxia through suppressing the role of potassium channels in BOO model rats.

Effect of filling rate on cystometric parameters in young and middle aged mice.

OBJECTIVESTo investigate the effect of changing the bladder filling rate during cystometry in younger (2–3 months) and older (13–14 months) C57BL/6J male mice.METHODSCystometry was performed on mice under anesthesia. Voiding cycles were established in each mouse at a pump delivery rate of 17 μl/min. After 30 min, the rate was increased sequentially to 25, 33, 41 and 49 μl/min. Each rate was maintained for 30 min. The following cystometric parameters were quantified: peak pressure amplitude, intercontractile interval (ICI), compliance, micturition pressure threshold and voiding efficiency.RESULTSBladder weights were significantly greater in older mice (42 mg vs. 27 mg, P < 0.01), but functional capacities were not different. The pressure amplitudes did not change as filling rate increased, nor did they differ between the 4-month and 13-month-old males. ICIs were not significantly different between young and mature mice. However, both groups exhibited a non-linear reduction in ICI with increasing filling rate, best described by a power curve (R2 > 0.93). Compliance was higher in the older mice at low filling rates (17 and 25 μl/min) but this difference diminished at higher rates. Compliance decreased with increasing flow rate in a non-linear manner, again with greater effects at low filling rates. Micturition pressure thresholds increased with increasing flow rate in a linear manner and older mice began voiding at higher pressures than younger. Both young and old mice exhibited voiding efficiencies of ~70%.CONCLUSIONSThe rate of volume delivery has complex effects on the timing of voiding and compliance. These findings argue for greater standardization of cystometry protocols and further investigation into afferent signaling to higher centers at different filling rates.

Protective Effects of Antimuscarinics on the Bladder Remodeling After Bladder Outlet Obstruction

Background/Aims: Overactive bladder associated with bladder outlet obstruction (BOO) is a highly prevalent condition, which is usually treated with antimuscarinics. However, the potential effects of antimuscarinics on the structure and function of bladder have not been investigated thus far. Methods: Sprague-Dawley(R) rats accepted bladder neck obstruction surgery or sham surgery, and then received treatment of three different antimuscarinics (Solifenacin, Darifenacin, and Tolterodine) or vehicle. After 3, 6 and 12 weeks, the bladder function and structure were measured. The effect of antimuscarinics on cellular alteration in vitro was observed under mechanical stimulation. Bladder morphology were examined by immunohistochemistry, and the bladder function were investigated by cystometry and strip contractility test. The expression of muscarinic receptors and inflammatory cytokines were measured by PCR and Western blotting. Results: Here we demonstrate, both in vitro and in vivo, that antimuscarinics are protective regulators for the bladder structure and function. Antimuscarinics decrease the weight of bladders with BOO. Antimuscarinics improve the voiding parameter and enhance the contraction of bladder smooth muscle. The results also show that antimuscarinics inhibit the proliferation of bladder smooth muscle cells both in vivo and in vitro, it can reduce the collagen deposition and inflammatory cytokines in bladders with BOO. During this process, the expression of M2 and M3 receptors was altered by antimuscarinics. Conclusion: Antimuscarinics could reverse the structural and functional changes of BOO bladder wall at cellular and tissue level, and the alteration of M2 and M3 receptors may be involved in this biological process.

AB287. SPR-14 High fat diet leads to impaired mitochondrial respiration, increased hydrogen peroxide emission and greater nerve-mediated purinergic contraction in mouse bladder

ObjectiveChronic high fat diet (HFD) is associated with the development of metabolic syndrome and type 2 diabetes, known risk factors for lower urinary tract symptoms, overactive bladder, and incontinence. HFD induced increases in oxidative stress and impaired mitochondrial integrity is a possible mechanism underlying this pathology. Our objectives were to determine bladder function following long-term HFD and characterize mitochondrial oxidant emission and respiratory function. We hypothesized that chronic HFD would lead to increased cholinergic mediated contraction and impaired mitochondrial respiration.MethodsMale C57BL/6J mice were maintained on a control diet (CD: 4% fat, 60% carbohydrates, 26% protein) or a HFD (45% fat, 35% carbohydrates, 20% protein) from 6 weeks of age for 20 weeks. A week prior to sacrifice, lower urinary tract function was assessed via void spot assay. Animals were euthanized and bladders and visceral fat depots were collected and weighed. Bladders were denuded of urothelium, cut into 8×2 mm strips and placed in tissue baths. Increasing concentrations of carbachol (cholinergic mediated) contraction and adrenergic relaxation to norepinephrine (NE) were measured. Electric field stimulated (EFS) contractions with and without atropine (cholinergic antagonist) were assessed. A separate group of bladders were denuded, homogenized and mitochondria quantified via citrate synthase activity. High-resolution respirometry measured oxygen consumption of individual electron transport chain complexes. Mitochondrial hydrogen peroxide (H2O2) emission was measured fluorimetrically using Amplex Ultra Red. Data were compared using a student t-test.ResultsHFD mice had a greater number of voids (P<0.005) and a larger void area (P<0.005). Blood glucose was greater in HFD (CD: 85±6.1; HFD: 139±5.9) as were body and overall visceral fat weights (P<0.0005). In contrast, HFD did not impact bladder weight. HFD did not change detrusor cholinergic contraction or adrenergic relaxation. However, detrusor contraction to EFS was markedly increased with HFD in both the presence and absence of atropine. Cholinergic inhibition blunted EFS contraction to a greater extent in CD bladders (CD: 75.7%±2.65; HFD: 42.3%±2.82, P<0.05) indicating that HFD had shifted EFS contraction to greater purinergic release. Citrate synthase activity indicated similar amounts of intact mitochondria. HFD resulted in a decreased respiratory capacity across 4 of 6 mitochondrial complexes investigated (P<0.05). Complex I and Complex II in state 4 conditions were unchanged. Additionally, mitochondrial H2O2 emission was increased in HFD bladders (CD: 11.4 pmols/min/mg ±0.53; HFD: 18.6±0.66, P<0.05).ConclusionsProlonged HFD leads to increased urinary frequency and volume, increased detrusor purinergic EFS contraction and mitochondrial dysfunction including decreased respiratory capacity and increased H2O2 emission. Further investigation of the role of mitochondrial dysfunction and resulting detrusor dysfunction following HFD is merited.Funding Source(s)None

Post-injury bladder management strategy influences lower urinary tract dysfunction in the mouse model of spinal cord injury.

To examine the effects of a different number of daily bladder squeezes on bladder dysfunction in mice with spinal cord injury (SCI). Spinal cord was transected at the Th8/9 in female C57BL/6N mice. Their bladders were manually squeezed to eliminate urine inside every day for 4 weeks. The mice were divided into three groups depending on the number of bladder squeezes; A: once daily, B: twice daily, C: three times daily. Four weeks after transection, single-filling cystometry were performed under an awake condition. NGF in the bladder mucosa and mRNA expression of P2X receptors and TRP channels in L6/S1 dorsal root ganglia (DRG) were measured. Bladder weight in group C was less than that of group A. Bladder capacity, post-void residual, and the number of non-voiding contractions during the storage phase were significantly larger in group A compared to group B or C. The level of NGF in groups C were lower compared to group A or B. The expression of P2X3 and TRPA1 in groups B and C was decreased compared to group A. The expression of P2X2 was decreased in groups B compared to group A. The post-injury bladder management after SCI with an increased number of daily bladder emptying improves the storage and voiding bladder dysfunction associated with the reduction of NGF in the bladder as well as P2X and TRP transcripts in lumbosacral DRG.

Non-invasive ultrasound measurements in male patients with LUTS and benign prostatic obstruction: implication for diagnosis and treatment

To evaluate the role of the ultrasound assessments of the bladder/detrusor wall thickness (BWT/DWT) and the ultrasound estimated bladder weight (UEBW) in lower urinary tract symptoms diagnosis and treatment. A systematic literature search from January 1995 until February 2016 was performed on PubMed using the following terms: "bladder weight", "bladder wall thickness", "detrusor wall thickness", "LUTS", "bladder outlet obstruction" (BOO), "benign prostatic obstruction". Additional references were obtained from the reference list of full-text manuscripts. Abstracts presented at the annual congresses of the European Association of Urology, American Urology Association and the International Continence Society were also included. BWT and DWT are accurate predictors of BOO. Despite several differences in the technique assessment, population characteristics and study design, the BWT/DWT accuracy for BOO was close to 90%. Furthermore, the ultrasound evaluation of bladder/detrusor thickness correlates positively with symptoms score and the main clinical parameters in BPH patients. Lastly, BWT and DWT have proved to be optimal predictor of response to medical/surgical treatment of lower urinary tract symptoms/benign prostatic obstruction (LUTS/BPO). However, the lack of standardization among the available studies marks these methods promising but still experimental and further studies are needed to clarify the standard methodology. The ultrasound evaluation of bladder/detrusor thickness appears to be simple, highly accurate and non-invasive technique to predict BOO and to evaluate the clinical outcomes after medical/surgical treatments for LUTS/BPH. The implementation of these techniques and their standardization will probably better define their role in the diagnostic algorithms of patients with LUTS and possibly reduce the number of unnecessary pressure flow-studies.

Evaluation of the effects of omega-3 & interferon alpha-2b administration on partial bladder outlet obstruction in a rat model.

Background & objectives:In bladder outlet obstruction-induced rat models, the transforming growth factor-beta (TGF-β) and collagen ratios have been shown to be increased. Increased TGF-β leads to fibrosis. In this study, the effect of omega-3 and interferon alpha-2b (IFN α-2b) was investigated on oxidative stress, inflammation and fibrosis in bladder structure in a partial bladder outlet obstruction (PBOO) rat model.Methods:A total of 35 male Wistar albino rats, weighing 300-350 g, were used in the study. The rats were randomly divided into five groups. At the end of the experimental period, bladders were harvested from all the rats, and pathological analysis of the rat bladder tissues was performed. In addition, investigations were carried out with enzymatic and non-enzymatic antioxidant systems to study the antioxidant properties of omega-3 fatty acid and IFN alpha-2b.Results:Increased bladder weight in the PBOO group, in comparison to the control group, was decreased by the administration of omega-3 and IFN α-2b (P=0.002). Significantly higher superoxide dismutase (SOD) levels were detected in group 2 in comparison to the control group. It was also detected that serum SOD, glutathione peroxidase and nitric oxide (NO) levels were significantly higher in group 2 when compared to the control group (P<0.05). In the pathologic evaluation, group 2 showed significantly increased inflammation and fibrosis compared to the control group. Omega-3 treatment significantly decreased inflammation. It was shown that IFN α-2b application partially decreased inflammation.Interpretation & conclusions:The results of the present study showed that in addition to the standard primary approaches to prevent the damage to the upper urinary tract secondary to PBOO, omega-3 fatty acid and IFN α-2b could be beneficial as adjunct treatment in clinical practice. However, this needs to be further investigated with prospective, randomized clinical trials with larger sample sizes.

Toll-Like Receptor 4 Activation Contributes to Diabetic Bladder Dysfunction in a Murine Model of Type 1 Diabetes.

Diabetic bladder dysfunction (DBD) is a common urological complication of diabetes. Innate immune system activation via Toll-like receptor 4 (TLR4) leads to inflammation and oxidative stress and was implicated in diabetes pathophysiology. We hypothesized that bladder hypertrophy and hypercontractility in DBD is mediated by TLR4 activation. Wild-type (WT) and TLR4 knockout (TLR4KO) mice were made diabetic by streptozotocin (STZ) treatment, and bladder contractile function and TLR4 pathway expression were evaluated. Immunohistochemistry confirmed the expression of TLR4 in human and mouse bladder. Recombinant high-mobility group box protein 1 (HMGB1) increased bladder TLR4 and MyD88 expression and enhanced contractile response to electrical field stimulation. Bladder expression of TLR4 and MyD88 and serum expression of HMGB1 were increased in STZ compared with control mice. Carbachol (CCh)-mediated contraction was increased in bladders from STZ mice, and TLR4 inhibitor CLI-095 attenuated this increase. Induction of diabetes by STZ in WT mice increased bladder weight and contractile responses to CCh and to electrical field stimulation. TLR4KO mice were not protected from STZ-induced diabetes; however, despite levels of hyperglycemia similar to those of WT STZ mice, TLR4KO STZ mice were protected from diabetes-induced bladder hypertrophy and hypercontractility. These data suggest that TLR4 activation during diabetes mediates DBD-associated bladder hypertrophy and hypercontractility.

Protective effects of the selective alpha1A-adrenoceptor antagonist silodosin against cyclophosphamide-induced cystitis in rats

We investigated the protective effects of a selective α1A-adrenoceptor antagonist, silodosin (Silod) on urinary bladder function in cyclophosphamide (CYP)-induced cystitis rats, with and without desensitization of the capsaicin (CAP)-sensitive afferent nerve pathway. Male Wistar rats (310–400 g) were pretreated with Silod (0, 100, or 300 μg/kg/day, p.o.) for 1 week before cystometry, and were administered either CYP (150 mg/kg, i.p.) or saline 2 days before the experiment. In another experiment, the rats were treated with CAP (125 mg/kg, s.c.) 4 days before the cystometry. The rat bladders were harvested, weighed, and evaluated histologically. The cystometric evaluation showed significant reductions in the intercontraction interval (ICI), single voided volume (SVV), and bladder compliance in CYP-treated rats compared to those in the vehicle-treated rats. High-dose Silod or CAP treatment significantly increased the ICI and SVV in the CYP rats. However, high-dose Silod treatment did not increase the ICI and SVV in CAP-treated CYP rats. Treatment with Silod did not improve the bladder weight, edema, and leukocyte infiltration resulting from the CYP-induced bladder inflammation. These data suggest that blockade of α1-adrenoceptors by Silod inhibited the CAP-sensitive afferent pathway in rats with cystitis.

Effect of Doxazosin on Autonomic Nervous Control and Urodynamics of Rat Urinary Bladder during Modeled Infravesical Obstruction.

The therapeutic effect of doxazosin (40 μg/kg/day over one month) on urinary bladder was examined in female rats with modeled chronic infravesical obstruction (IVO) produced by graduated mechanical constriction of the proximal urethral segment. In one month, IVO induced a pronounced vesical hypertrophy both in treated and untreated rats that manifested in increased bladder weight and capacity, the latter increment being pronouncedly greater in treated rats. In untreated IVO rats, infusion cystometry revealed elevated basal intravesical pressure of void bladder P0, markedly increased maximal (premicturitional) pressure Pmax, and increased amplitude of spontaneous oscillations of intravesical pressure ΔPdet in filled bladder. Doxazosin produced no significant effect on Pmax rise during IVO, but prevented elevation of P0 and increment of ΔPdet in filled bladder. During gradual filling of urinary bladder in control (intact) rats, the parasympathetic vesical influences increased progressively, while in untreated IVO rats, the adrenergic influences prevailed even at maximal filling of the bladder. In IVO rats, doxazosin prevented the bias of the sympathetic-parasympathetic balance in the filled bladder in favor of sympathetic influences, but did not prevent this bias in a void bladder. It is hypothesized that α-adrenoblockers improve micturition during IVO caused by benign prostatic hyperplasia not only by decreasing the urethral resistance to urine flow due to down-regulation of prostate smooth muscle tone, but also by a direct action of these blockers on detrusor adrenergic receptors and central structures involved in urinary bladder control.

Can Bladder and Prostate Sonomorphology Be Used for Detecting Bladder Outlet Obstruction in Patients With Symptomatic Benign Prostatic Hyperplasia?

To investigate the accuracy of the bladder and prostate sonomorphologic parameters for the diagnosis of bladder outlet obstruction (BOO) in patients with lower urinary tract symptoms (LUTS)/benign prostatic hyperplasia (BPH). First-visit men, aged ≥ 50 years, with LUTS/BPH were prospectively enrolled. Added to the initial basic evaluation, all patients underwent pelvic ultrasonography and pressure flow study. The pressure flow study was used as a reference standard for BOO, and according to its results, patients were divided into BOO and non-BOO groups. The sonomorphologic findings were compared between both groups, and the diagnostic accuracy of the significant parameters was determined. In total, 157 patients were included. Of these, 48 (30.57%) had BOO and 109 (69.43%) did not. Bladder wall thickness (BWT), ultrasound estimated bladder weight (UEBW), and intravesical prostatic protrusion (IPP) were the sonomorphologic parameters that differed significantly between both groups (P < .001). By receiver operating characteristic curve analysis, the optimal cutoff values distinguishing patients with BOO were BWT of 3.7 mm (area under the curve [AUC]: 0.940), UEBW of 31.5 g (AUC: 0.835), and IPP of 10.9 mm (AUC: 0.874). The sensitivity, specificity, and accuracy of BWT, UEBW, and IPP were 95.00%, 90.91%, and 93.55%; 75.44, 57.97%, and 65.62%; and 87.90%, 76.43%, and 82.17%, respectively. BWT, UEBW, and IPP measurements can be used to detect BOO in LUTS/BPH patients. The simple estimation of these sonomorphologic parameters by suprapubic approach makes these measurements potentially suitable methods to judge BOO noninvasively during the routine clinical evaluation of BPH.

Abstract 5241: Nitric oxide-releasing naproxen prevents muscle invasive bladder cancer

Abstract Bladder cancer is the second most common and a leading cause of death among genitourinary cancers worldwide. Particularly, untreated muscle invasive bladder cancer has high mortality (&amp;gt;85% patients) leading to death within 2 years of diagnosis. Preventing this deadly form is highly imperative since the currently available options to patients with invasive disease remained essentially unchanged and no effective drugs have been approved in past two decades. Several non-steroidal anti-inflammatory agents (NSAIDs) have shown promising chemopreventive activity in many cancers. The common adverse events with NSAIDs, especially gastrointestinal (GI) morbidities, including complications in both upper and lower GI tract drives the need for development of safer agents. To overcome this various nitric oxide (NO)-linked NSAIDs have been synthesized. Here we investigated the NO-releasing Naproxen (NO-Naproxen) with proven anti-inflammatory and GI protecting effects for its efficacy in preventing bladder cancer. Transgenic mouse model (UPII-SV40T; n = 30/group) that develop muscle invasive urothelial tumors were generated, genotyped and fed modified AIN-76A diet containing NO-naproxen (0, 300 and 600 ppm) starting at 6 weeks of age. At 40 weeks age, control (0 ppm) and experimental diet (300 and 600 ppm) fed mice were euthanized and urinary bladders were analyzed. Control diet fed male and female transgenic mice developed high grade, invasive transitional cell carcinoma (TCC) of bladder resulting in significant increase in bladder weights (140.2±9.8 mg; p&amp;lt;0.0001 and 34.2+0.8 mg; p&amp;lt;0.0001 respectively) compared with wild type mice (27.3±0.8 mg and 14.8±0.53 mg). These tumors had a significant disregulation of proliferation, cell cycle markers and antioxidant enzymes similar to human tumors. NO-Naproxen administered mice had normal body weight gain; and gross tissue analysis and showed no signs of overt toxicities. Treatment of transgenic mice with NO-Naproxen led to significant suppression of bladder weight in both genders (up to 58% in males, p&amp;lt;0.0001; up to 21% in females, p&amp;lt;0.005) compared to control group. While there was no dose-dependent increase in tumor inhibition, mice on NO-Naproxen diet had developed significantly less muscle invasive tumors suggesting inhibitory effect of treatment on disease progression. Urothelial tumor progression to invasive TCC was inhibited in both male (up to 54%; p&amp;lt;0.005) and females mice (up to 85%; p&amp;lt;0.0001) of the experimental diet groups. Molecular analysis of urothelial tumors via real-time PCR, IHC and/or western blotting showed inhibitory effect of NO-Naproxen on proliferation and inflammatory markers (PCNA, Cyclins, COX2, and IL1b) and showed modulation of antioxidant enzymes (CAT, GPx, GST, NQO1, and SOD3). Our results suggest that NO-Naproxen may be a promising agent for preventing urothelial TCC and warrants further investigation. (Supported in part by NCI-CN53300) Citation Format: Venkateshwar Madka, Altaf Mohammed, Qian Li, Yuting Zhang, Stanley Lightfoot, Xue-Ru Wu, Vernon Steele, Levy Kopelovich, Chinthalapally V. Rao. Nitric oxide-releasing naproxen prevents muscle invasive bladder cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5241.

Hydronephrosis and Urinary Bladder Hypertrophy in ROMK Bartter's Mouse

The renal outer medullary potassium channel (ROMK; Kir1.1) plays an important role in Na+ and K+ homeostasis. ROMK knockout mice (KO) show a similar phenotype to Bartter's syndrome of salt wasting and dehydration due to reduced Na‐2Cl‐K‐cotransporter activity. ROMK KO mice also show hydronephrosis; however, the mechanism of this phenotype has not been understood. We investigated ROMK expression and regulation by high and low K intake in mouse bladder, by Western blot and immunofluorescence (IF) staining. We also compared the bladder weight, bladder capacity, and thickness of urothelium and smooth muscle layer between WT and ROMK KO mice. Western blotting detected ROMK expression at 45~48 kDA, and IF staining showed that the luminal membrane of umbrella cells were strongly stained in WT compared to KO mice. The expression of ROMK in the luminal membrane disappeared and signal intensity of the detrusor muscle was decreased by low potassium diet (LK) in WT mice. In contrast, there was no significant change in ROMK intensity, but the thickness of urothelium was increased after mice were treated with HK diet. The thickness of urothelium was 168.1±25.8 μm in normal K (NK) and was 300.7±83.6 μm (p&lt;0.001) in the HK treated group compared with NK in WT mice at the same age. The bladder weight, bladder capacity and the thickness of urothelium in ROMK KO showed average increases 2~4 fold greater than WT at both 2 months and 12 months of age. The thickness of the urothelium was 648.8±33.2 μm vs 302.7±16.5 μm (p&lt;0.001) and the detrusor muscle 1940.7±98.9 μm vs 1308.2±102.1 μm (p=0.013) respectively in 12‐month ROMK KO mice. In conclusion, ROMK expressed in the urinary bladder can be regulated by K+ intake; significant enlargement and hypertrophy of the bladder may contribute to hydronephrosis in ROMK KO mice.Support or Funding InformationNIH RO1 DK099284 and PO1 DK 17433

MP68-20 LUMBAR TO SACRAL ROOT REROUTING TO RESTORE VOIDING FUNCTION IN A FELINE SPINAL CORD INJURY MODEL: URODYNAMIC AND RETROGRADE NERVE TRACING RESULTS FROM A PILOT STUDY

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology II1 Apr 2016MP68-20 LUMBAR TO SACRAL ROOT REROUTING TO RESTORE VOIDING FUNCTION IN A FELINE SPINAL CORD INJURY MODEL: URODYNAMIC AND RETROGRADE NERVE TRACING RESULTS FROM A PILOT STUDY Ornella Lam van ba, Mary F. Barbe, Romain Caremel, Shachar Aharony, Oleg Loutochin, Line Jacques, Matthew Woods, Gerald F. Tuite, Michael R. Ruggieri, Lysanne Campeau, and Jacques Corcos Ornella Lam van baOrnella Lam van ba More articles by this author , Mary F. BarbeMary F. Barbe More articles by this author , Romain CaremelRomain Caremel More articles by this author , Shachar AharonyShachar Aharony More articles by this author , Oleg LoutochinOleg Loutochin More articles by this author , Line JacquesLine Jacques More articles by this author , Matthew WoodsMatthew Woods More articles by this author , Gerald F. TuiteGerald F. Tuite More articles by this author , Michael R. RuggieriMichael R. Ruggieri More articles by this author , Lysanne CampeauLysanne Campeau More articles by this author , and Jacques CorcosJacques Corcos More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1357AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Following complete spinal cord injury (SCI), lumbar to sacral rerouting surgery has the potential to allow voiding via a new skin-central nervous system-bladder reflex pathway. We aimed to assess whether this surgery is effective in treating neurogenic bladder in a feline SCI model METHODS Eight cats underwent SCT at T9-T10. Unilateral L7-S1 ventral root anastomosis was performed 1 month later in 6 cats. Remaining dorsal and ventral roots below L7 were transected, bilaterally. Bladders were checked three times per day and emptied with the Crede maneuver. The new neurological pathway was evaluated at 3, 5, 7 and 9 months by electrical and manual cutaneous stimulation of L7 dermatomes and urodynamics. At 9 months, cats were euthanized 2 weeks after bladder wall Fluorogold retrograde tracer injection and an assessor-blinded pathological exam of the L7-S1 root anastomosis site and lumbosacral cord ventral horns for presence of Fluorogold labeled cell bodies was performed. Outcomes included: 1) urodynamic responses to cutaneous or electrical stimulation the L7 dermatome 2) axonal growth from L7 and S1 ventral horns to the bladder; and 3) incidence of bladder hypertrophy RESULTS At 9 months, 3 of the 6 rerouted cats exhibited increased detrusor pressure provoked by unilateral cutaneous stimulation, and one cat bilaterally. Ipsilateral cutaneous stimulation produced a voiding stream in 2 cats. SCI controls did not respond. Mean bladder volume or weight was not different between rerouted animals and SCI controls. Postmortem studies determined the anastomosed ventral roots were one-sided L7 to S1 (3 right-sided rerouting and 3 left-sided). One right-rerouted cat also showed the left L7 ventral root was annealed to the right S1 root. Although some aberrant axonal regrowth was observed, all 6 rerouted animals showed regrowth of axons from the L7 ventral horn segment to the bladder using retrograde tracing, (Figure 1) including the animal with bilateral input to S1 CONCLUSIONS L7-to-S1 ventral root rerouting surgery below the level of SCT showed successful axonal regrowth to the bladder in all rerouted animals and induced urodynamic responses to cutaneous stimulation in a subset. This feasibility study paves the way for future studies for bladder re-innervation © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e897 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Ornella Lam van ba More articles by this author Mary F. Barbe More articles by this author Romain Caremel More articles by this author Shachar Aharony More articles by this author Oleg Loutochin More articles by this author Line Jacques More articles by this author Matthew Woods More articles by this author Gerald F. Tuite More articles by this author Michael R. Ruggieri More articles by this author Lysanne Campeau More articles by this author Jacques Corcos More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

High Sensitivity of an Ha-RAS Transgenic Model of Superficial Bladder Cancer to Metformin Is Associated with ∼240-Fold Higher Drug Concentration in Urine than Serum.

While pharmacoepidemiologic and laboratory studies have supported the hypothesis that the antidiabetic drug metformin may be useful in treating or preventing cancer, there is limited evidence to suggest which specific cancer sites may be particularly sensitive. Sensitivity likely is determined both by features of tumor pathophysiology and by pharmacokinetic factors. We used UPII-mutant Ha-ras transgenic mice that develop hyperplasia and low-grade, papillary urothelial cell carcinoma to determine whether metformin has activity in a model of superficial bladder cancer. Metformin significantly improved survival, reduced urinary tract obstruction, reduced bladder weight (a surrogate for tumor volume), and led to clear activation of AMP α kinase and inhibition of mTOR signaling in neoplastic tissue. We investigated the basis of the unusual sensitivity of this model to metformin, and observed that following oral dosing, urothelium is exposed to drug concentrations via the urine that are approximately 240-fold higher than those in the circulation. In addition, we observed that bladder cancer cell lines (RT4, UMUC-3, and J82) with homozygous deletion of either TSC1 or PTEN are more sensitive to metformin than those (TEU2, TCCSUP, and HT1376) with wild-type TSC1 and PTEN genes. Our findings provide a strong rationale for clinical trials of oral metformin in treatment of superficial bladder cancer.

In Utero and Lactational TCDD Exposure Increases Susceptibility to Lower Urinary Tract Dysfunction in Adulthood.

Benign prostatic hyperplasia, prostate cancer, and changes in the ratio of circulating testosterone and estradiol often occur concurrently in aging men and can lead to lower urinary tract (LUT) dysfunction. To explore the possibility of a fetal basis for the development of LUT dysfunction in adulthood, Tg(CMV-cre);Nkx3-1(+/-);Pten(fl/+) mice, which are genetically predisposed to prostate neoplasia, were exposedin uteroand during lactation to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 1 μg/kg po) or corn oil vehicle (5 ml/kg) after a single maternal dose on 13 days post coitus, and subsequently were aged without further manipulation, or at 8 weeks of age were exposed to exogenous 17 β-estradiol (2.5 mg) and testosterone (25 mg) (T+E2) via slow release subcutaneous implants.In uteroand lactational (IUL) TCDD exposure in the absence of exogenous hormone treatment reduced voiding pressure in adult mice, but otherwise had little effect on mouse LUT anatomy or function. By comparison, IUL TCDD exposure followed by exogenous hormone treatment increased relative kidney, bladder, dorsolateral prostate, and seminal vesicle weights, hydronephrosis incidence, and prostate epithelial cell proliferation, thickened prostate periductal smooth muscle, and altered prostate and bladder collagen fiber distribution. We propose a 2-hit model whereby IUL TCDD exposure sensitizes mice to exogenous-hormone-induced urinary tract dysfunction later in life.