Bladder Weight Research Articles

Changes of neuregulin-1(NRG-1) expression in a rat model of overactive bladder induced by partial urethral obstruction: is NRG-1 a new biomarker of overactive bladder?

BackgroundTo determine whether neuregulin-1(NRG-1) is a potential new biomarker of overactive bladder (OAB) induced by partial urethral obstruction in a rat model of OAB and to evaluate the urothelium as a therapeutic target of OAB.MethodsFemale Sprague–Dawley rats were separated into three 20-animal groups: normal, OAB, and 5-hydroxymethyl tolterodine (5-HMT)-treated OAB. In the OAB and OAB + 5-HMT groups, the urethra of each animal was partially obstructed; the OAB + 5-HMT group received intravenous 5-HMT for 3 weeks. At the conclusion of the 5-HMT dosing, the rats in each group underwent cystometrography, and the bladders were histologically evaluated. The expression of brain derived-neurotrophic factor (BDNF) and NRG-1 were evaluated in the urothelium.ResultsCompared with the control group, the OAB group showed a markedly increased bladder weight and a significant decrease in the micturition interval and volume; rats in the OAB + 5-HMT group showed decreased bladder weights and an improved micturition interval and volume. BDNF and NRG-1 were expressed at significantly higher levels in the OAB group, and were significantly reduced in the OAB + 5-HMT group compared with the control group.ConclusionsThe study suggests that NRG-1 is a potential new biomarker of OAB; the urothelium might be a therapeutic target for OAB treatment.

Synergistic effect of melamine in combination with cyanuric acid on urinary tract toxicity in rats

The present study investigated the urinary tract toxicity of melamine alone or in combination with cyanuric acid in rats. Male rats were orally administered melamine alone (800 mg/kg) or melamine plus cyanuric acid (50 mg/kg each) for 3 consecutive days. Although melamine treatment did not show any significant difference in body weight, kidney or urinary bladder weight, serum blood urea nitrogen (BUN) or creatinine levels, it caused a decrease in urinary pH and mild histopathological alterations in the kidney and urinary bladder. In contrast, co-administration of melamine and cyanuric acid induced a decrease in body weight, an increase in kidneys and urinary bladder weights, and an elevation in serum BUN and creatinine levels, which were not observed in animals treated with melamine alone. Histopathological examination showed that the incidence and severity of histopathological lesions in the kidney and urinary bladder were much higher than those in the melamine group. Urinalysis revealed an increase in urine occult blood, leukocytes and protein and a decrease in urinary pH. These results indicate that concomitant administration of cyanuric acid synergistically potentiated the urinary tract toxicity induced by melamine in rats. In this study, we first demonstrated the synergistic toxic effects of melamine in combination with cyanuric acid on urinary bladder and urinalysis in rats.

Limitaciones de los fármacos dependientes de insulina para el tratamiento de la diabetes mellitus tipo 2

In this study, we review the efficacy and safety limitations of insulin-dependent oral antidiabetic agents. In terms of efficiency, the main drawback of metformin, sulfonylureas, gliptins and –to a lesser extent–glitazones is durability. No drug per se is able to maintain stable blood glucose control for years.Metformin, sulfonylureas and gliptins have demonstrated safety. Experience with the first two drug groups is more extensive. The main adverse effect of metformin is gastrointestinal discomfort. Major concerns related to the use of sulfonylureas are hypoglycemia and weight gain. The use of pioglitazone has been associated with an increased risk of bladder cancer, edema, heart failure, weight gain, and distal bone fractures in postmenopausal women. The most common adverse reactions associated with glucagon-like peptide-1 agonists are gastrointestinal discomfort that sometimes leads to treatment discontinuation.

Subchronic Toxicity Evaluation of Anthraquinone in Fischer 344 Rats

Female F344 rats were exposed to anthraquinone (AQ) by dietary feed at concentrations of 0, 50, 150, 469, 938, 1875, or 3750 ppm for 2 or 13 weeks. End points evaluated included clinical observations, body weights, serum chemistry, blood AQ, gross pathology, organ weights, and select tissue histopathology. Mean body weight and food consumption were 5% to 10% lower than control values in rats of the ≥938 ppm group during study weeks 2 through 13. Occasional decreases in body weight means were also observed in rats of the 150 and 469 ppm groups. Increases in liver, kidney, and spleen weights were observed in rats exposed to AQ diet concentrations ≥150 ppm for 13 weeks. Urinary bladder weights were increased at ≥469 ppm. Liver and spleen weights were also increased following 2 weeks of exposure. Liver weight increases were clearly dependent on AQ concentration. At 2 weeks, decreases in serum aspartate aminotransferase (AST), blood urea nitrogen, and creatinine concentrations were observed in higher AQ exposure groups, and AST was decreased at 13 weeks (≥1875ppm). Microscopic alterations were observed in the liver (mild centrilobular hypertrophy), spleen (mild hematopoietic cell proliferation and pigmentation), and kidneys (minimal hyaline droplets) of rats exposed to AQ for 13 weeks. Blood AQ concentrations ranged from 0.75 to 14.8 µg/mL in rats of the 150 to 3750 ppm groups, respectively, and were similar in value following either 2 weeks or 13 weeks of exposure. A no observed adverse effect level of 469 ppm AQ (31.3 mg/kg/d) was selected based on the absence of liver histopathology.

P53-stabilizing Agent CP-31398 Prevents Growth and Invasion of Urothelial Cancer of the Bladder in Transgenic UPII-SV40T Mice

The high prevalence of bladder cancer and its recurrence make it an important target for chemoprevention. About half of invasive urothelial tumors have mutations in p53. We determined the chemopreventive efficacy of a p53-stabilizing agent, CP-31398, in a transgenic UPII-SV40T mouse model of bladder transitional cell carcinoma (TCC) that strongly resembles human TCC. After genotyping, six-week-old UPII-SV40T mice (n = 30/group) were fed control (AIN-76A) or experimental diets containing 150 or 300 ppm of CP-31398 for 34 weeks. Progression of bladder cancer growth was monitored by magnetic resonance imaging. At 40 weeks of age, all mice were killed; urinary bladders were collected to determine weights, tumor incidence, and histopathology. There was a significant increase in bladder weights of transgenic versus wild-type mice (male: 140.2 mg vs 27.3 mg, P < .0001; female: 34.2 mg vs 14.8 mg, P < .0001). A significant decrease in the bladder tumor weights (by 68.6–80.2%, P < .0001 in males and by 36.9–55.3%, P < .0001 in females) was observed in CP-31398-treated mice. Invasive papillary TCC incidence was 100% in transgenic mice fed control diet. Both male and female mice exposed to CP-31398 showed inhibition of invasive TCC. CP-31398 (300 ppm) completely blocked invasion in female mice. Molecular analysis of the bladder tumors showed an increase in apoptosis markers (p53, p21, Bax, and Annexin V) with a decrease in vascular endothelial growth factor in transgenic mice fed CP-31398. These results suggest that p53-modulating agents can serve as potential chemopreventive agents for bladder TCC.

Sacral Foraminal Osteochondroma Causing Radiculopathy

Osteochondromas are the most common benign tumor of bone. These tumors are composed of a histologically normal osseous projection and a cartilaginous cap. The lesion is commonly metaphyseal, likely arising from a physis. Clinically, osteochondromas usually present as a painless protrusion or bump. The most common age of presentation is between ten and twenty years of age. These tumors are rarely found in the spinal column1. Only 1% to 4% of primary osteochondromas occur in the spine2,3 and, of these, less than 0.5% originate in the sacrum4. We present a case of an anterior sacral osteochondroma that extended into the neural foramen and caused radiculopathy in a young athletic man. Although sacral osteochondromas have been previously reported, this case demonstrates the ability of these tumors to extend into and through the neural foramen, producing radiculopathy. The patient was informed that data concerning the case would be submitted for publication, and he provided consent. ### Clinical History and Diagnosis An otherwise healthy twenty-year-old male recreational soccer player presented with the symptom of six months of right leg pain. The history was noteworthy for prior back pain from a mild lumbar disc herniation on the right at the L4-L5 level, diagnosed with magnetic resonance imaging (MRI), which had been managed conservatively. For the six months prior to presentation, he reported experiencing a tingling and stabbing pain in the right buttock, posterior thigh, calf, and lateral aspect of the foot. He also had new onset of a “pins-and-needles” sensation in the lateral aspect of the right foot. The symptoms worsened with activity and prolonged sitting on hard surfaces. The patient stopped playing soccer secondary to severe stabbing leg pain. He denied muscle weakness, saddle anesthesia, bowel or bladder incontinence, weight loss, or night pain. Initial treatment was with physical therapy, nonsteroidal …

Macrophage migratory inhibitory factor promotes bladder cancer progression via increasing proliferation and angiogenesis

Macrophage migratory inhibitory factor (MIF) is a proinflammatory cytokine shown to promote tumorigenesis. Using the N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) model of bladder cancer, we previously showed that MIF knockout mice display decreased angiogenesis and invasion compared with wild-type. This study examines the role of MIF in bladder cancer via use of oral inhibitors of MIF. In vitro, high-grade bladder cancer cells were treated with recombinant human MIF +/- (rhMIF+/-) inhibitor. Measurements included cell counts, proliferation by (3)H-thymidine incorporation (TdR), extracellular signal-regulated kinase (ERK) phosphorylation by western blot analysis, messenger RNA (mRNA) expression by quantitative PCR and protein secretion by enzyme-linked immunosorbent assay. Treatment with rhMIF increased ERK phosphorylation, cell counts, TdR and mRNA expression and protein secretion of vascular endothelial growth factor, which were blocked by specific inhibitors of ERK and MIF. In vivo, 3-month-old male C57Bl/6 mice were given BBN for 22 and 16 weeks in study 1 and study 2, respectively. Mice (n = 8-10 per group) were gavaged with vehicle or doses of MIF inhibitors daily from weeks 16-22 in both studies. Average bladder weights, reflecting tumor mass, tumor stage/burden, mitotic rate and proliferation indices, and microvessel densities were reduced in inhibitor groups versus controls. In summary, MIF promotes bladder cancer via increasing cell proliferation and angiogenesis and oral inhibitors of MIF may prove useful in treatment of this disease.

Effect of ovariectomy, 17-beta estradiol, and progesterone on histology and estrogen receptors of bladder in female partial bladder outlet obstruction rat model

The aim of this study was to investigate the effect of bilateral ovariectomy (OVX), 17-beta estradiol (E2), and progesterone (P4) on the histology and estrogen receptor (ER) expression of the bladder using a female partial bladder outlet obstruction (pBOO) rat model. A total of 60 female Sprague-Dawley rats were evenly assigned into six groups of 10 each. Group A served as the control. Groups B-F underwent induced pBOO. Groups C-F underwent OVX. Groups D-F were given E2 (0.1 mg/kg/day), Group E was given P4 (1 mg/kg/day), and Group F was given P4 and dehydroepiandrosterone (DHEA) (300 μg/kg/day) by an Alzet pump. Four weeks later, serum E2 and P4 levels were evaluated. Each rat was anesthetized and the urinary bladder was removed for weighing and histological study. Expression of ER-β was not significantly different between the control group and the other study groups. pBOO was shown to increase both bladder weight and detrusor muscle thickness. OVX had an additive effect to BOO on increased blood vessel density in the bladder. E2 was shown to increase blood vessel density, while P4 supplementation decreased blood vessel density. DHEA did not cause any significant effects on blood vessel density. Hormone therapy did not change the expression of ER in bladder outlet obstruction. Estradiol stimulated the increased angiogenesis of the bladder detrusor but P4 decreased the angiogenesis of the bladder detrusor. DHEA had no effect on the bladder detrusor.

Gap junction expression and the effects of gap junction inhibitors in overactive bladder models: does ovariectomy have a role?

In this study, gap junction expression and the effects of estrogen deficiency and gap junction inhibitors were investigated in overactive bladder models which were created by bladder outlet obstruction. In our study, we created four groups as control, ovariectomy, bladder outlet obstruction (BOO), and ovariectomy + BOO. We investigated the effects of oxybutynin and 18-alpha glycyrrhetinic acid (18-α-GA) which is a gap junction blocker on isolated detrusor strips. Western blot method was used to measure the level of connexin-43 in detrusor. Bladder weights were significantly increased in the BOO and ovariectomy + BOO groups (p < 0.05). There was no statistically significant difference in the maximal contraction responses to carbachol between ovariectomy and control groups. In BOO and ovariectomy + BOO groups, contractile responses were significantly prominent with higher doses of carbachol. Oxybutynin-induced relaxant responses of BOO and ovariectomy + BOO groups were significantly higher than control group (p < 0.05). The relaxation effect of 18-a-GA was more effective in the obstruction groups. Among those two groups, the relaxation observed in BOO group was higher than ovariectomy + BOO group in higher doses of 18-a-GA. Connexin-43 expression was increased in BOO group compared with the control group (p = 0.006). Ovariectomy did not change connexin-43 expression alone; however, when combined with BOO, connexin-43 expression decreased significantly (p = 0.023). Ovariectomy had no effect on the gap junctions in the bladder and bladder overactivity alone. Therefore, obstruction is the main factor that increases the amount of gap junctions, and gap junction blockers are thus more effective in obstruction. However, ovariectomy was shown to decrease the expression of gap junctions and relaxation effect of gap junction blockers, when combined with BOO.

Chemoprevention of BBN-Induced Bladder Carcinogenesis by the Selective Estrogen Receptor Modulator Tamoxifen

Bladder cancer is the fifth most frequent tumor in men and ninth in women in the United States. Due to a high likelihood of recurrence, effective chemoprevention is a significant unmet need. Estrogen receptors (ERs), primarily ERβ, are expressed in normal urothelium and urothelial carcinoma, and blocking ER function with selective ER modulators such as tamoxifen inhibits bladder cancer cell proliferation in vitro. Herein, the chemoprotective potential of tamoxifen was evaluated in female mice exposed to the bladder-specific carcinogen, N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). Carcinogen treatment resulted in a 76% tumor incidence and increased mean bladder weights in comparison to controls. In contrast, mice receiving tamoxifen concurrent (8–20 weeks) or concurrent and subsequent (8–32 weeks) to BBN administration had no change in bladder weight and only 10% to 14% incidence of tumors. Non-muscle-invasive disease was present in animals treated with tamoxifen before (5–8 weeks) or after (20–32 weeks) BBN exposure, while incidence of muscle-invasive bladder carcinoma was reduced. ERβ was present in all mice and thus is a potential mediator of the tamoxifen chemoprotective effect. Surprisingly, ERα expression, which was detected in 74% of the mice exposed to BBN alone but not in any controlmice, was correlated with tumor incidence, indicating a possible role for this receptor in carcinogen-induced urothelial tumorigenesis. Thus, these data argue that both ERα and ERβ play a role in modulating carcinogen-induced bladder tumorigenesis. Administration of tamoxifen should be tested as a chemopreventive strategy for patients at high risk for bladder cancer recurrence.

SU‐E‐T‐653: Predicting Objective Function Weights for IMRT Prostate Treatment Planning Using Patient Anatomy

Purpose: To develop a prediction model for objective function weights for intensity‐modulated radiation therapy (IMRT) prostate treatment planning with multiple objectives using geometry information from patient anatomy. Methods: A previously developed inverse optimization method (IOM) was used to reverse‐engineer optimal objective function weights (inverse weights) from an observed treatment plan. We developed a regression model to predict the weights for IMRT prostate treatment planning using patient anatomy from 25 patients. The ratio of the overlap volumes of the rectum and bladder with the planning target volume expanded by 1cm was used to predict the bladder and rectum weights. The femoral head weights were included in the model as a small fixed weight (1%). The model was validated using leave‐one‐out cross‐validation. We evaluated the model by comparing the treatment plans generated through inverse planning using the inverse weights from IOM and the predicted weights from the regression model. Results: On average, V54Gy for the bladder was 36.1% using the inverse weights and 36.6% using the predicted weights. V70Gy for the bladder was 23.2% (inverse) and 23.5% (predicted). For the rectum, V54Gy was 34.6% (inverse) and 33.9% (predicted), and V70Gy was 22.6% (inverse) and 22.3% (predicted). For each criterion, the difference between the inverse and predicted metrics was not statistically significant. All treatment plans from the predicted weights satisfied the clinical criteria. Conclusion: Our results show that objective function weights are well‐predicted by the regression model. This approach may support the genesis of personalized weights in IMRT treatment planning. This research was supported in part by the Natural Sciences and Engineering Research Council of Canada (NSERC) and Ontario Graduate Scholarship (OGS).

Myocardin and microRNA‐1 modulate bladder activity through connexin 43 expression during post‐natal development

Overactive bladder (OAB) is a pervasive clinical problem involving alterations in both neurogenic and myogenic activity. While there has been some progress in understanding neurogenic inputs to OAB, the mechanisms controlling myogenic bladder activity are unclear. We report the involvement of myocardin (MYOCD) and microRNA-1 (miR-1) in the regulation of connexin 43 (GJA1), a major gap junction in bladder smooth muscle, and the collective role of these molecules during post-natal bladder development. Wild-type (WT) mouse bladders showed normal development from early post-natal to adult including increases in bladder capacity and maintenance of normal sensitivity to cholinergic agents concurrent with down-regulation of MYOCD and several smooth muscle cell (SMC) contractile genes. Myocardin heterozygous-knockout mice exhibited reduced expression of Myocd mRNA and several SMC contractile genes concurrent with bladder SMC hypersensitivity that was mediated by gap junctions. In both cultured rat bladder SMC and in vivo bladders, MYOCD down-regulated GJA1 expression through miR-1 up-regulation. Interestingly, adult myocardin heterozygous-knockout mice showed normal increases in bladder and body weight but lower bladder capacity compared to WT mice. These results suggest that MYOCD down-regulates GJA1 expression via miR-1 up-regulation, thereby contributing to maintenance of normal sensitivity and development of bladder capacity.

Bladder function in 17β-estradiol-induced nonbacterial prostatitis model in Wister rat

To investigate bladder function in a model of nonbacterial prostatitis (NBP) induced in castrated rats by 17β-estradiol injection. Ten-month-old male Wistar rats were divided into two groups, sham and NBP (both N = 8). NBP was induced by castration followed by daily subcutaneous injection of 17β-estradiol for 30 days. On the 31st day after surgery, we investigated (1) voiding behavior, (2) bladder blood flow (BBF), (3) prostate and bladder weight, and proinflammatory cytokines (TNF-α and CXCL1) levels and (4) bladder contractile responses to electrical field stimulation (EFS), carbachol and KCl. (1) Voiding behavior (average micturition volume, total urine volume and number of micturitions) and (2) BBF were not significantly different between the sham and NBP groups. (3) NBP led to a significant decrease in prostatic weight and increase in proinflammatory cytokine levels in the prostate, but NBP did not cause a significant change in bladder weight or proinflammatory cytokine levels in the bladder. (4) Bladder contractile forces in response to EFS, carbachol and KCl were not significantly affected by NBP. In this rat model, NBP did not cause a significant change in the level of proinflammatory cytokines in the bladder and affect bladder function.

Abstract 3690: p53 stabilizing agent CP-31398 prevents urinary bladder tumor growth and invasion in transgenic UPII-SV40T mice.

Abstract The United States will have approximately 73,510 new cases of bladder cancer in 2012 with 14,880 deaths. More than 50% of the invasive (lamina and muscularis) bladder tumors have mutations in p53 and/or pRB. p53 appears to play a major role in the etiology of human bladder cancer, particularly in the high grade invasive type. Structural and functional defects in p53 are found in majority of human transitional cell carcinomas (TCC). The high prevalence of bladder cancer and the frequency of tumor recurrence make it an important target for chemoprevention. In UPII-SV40T transgenic mice, induced bladder TCC bears strong resemblance to that of human TCC in phenotype and in the mode of progression. In the present study, we determined the chemopreventive efficacy of a p53 stabilizing agent, CP-31398, using transgenic UPII-SV40T mouse model. After genotyping, six-week old UPII-SV40T mice (n=32/group) were fed control (AIN-76A) or experimental diets containing 150 or 300 ppm CP-31398 for ∼34 weeks. Progression of bladder cancer was monitored by MRI imaging. At 40 weeks of age, all mice were euthanized; urinary bladders were collected to determine weights, tumor incidence and multiplicity; and histopathology. There was a significant increase in bladder weights of transgenic vs. wild type mice (male 140.2 mg vs. 27.3 mg, p&amp;lt;0.0001; female 34.2 mg vs, 14.8 mg p&amp;lt;0.0001, respectively). A significant decrease in the tumor growth as evidenced by decreased bladder weights (by 52-65%, p&amp;lt;0.000 in males; by 32 %, p&amp;lt;0.001 in females) was observed in CP-31398 treated mice. Invasive papillary TCC incidence was 100 % in transgenic mice fed with control diet. The mice exposed to 150 and 300 ppm CP-31398 showed dose dependent inhibition (29%, p&amp;lt;0.01; 57%, p&amp;lt;0.005) of invasive TCC with 28% and 53% carcinoma In situ, respectively, suggesting suppression of invasive tumor growth. Kidney abnormalities like ureter obstruction and nephrohydrosis were observed in almost all control mice but significantly less frequent in CP-31398-fed mice. Molecular analysis of the bladder tumors showed an increase in apoptosis markers (p53, p21, Bax) with a decrease in angiogenic markers (CD31 and VEGF) in transgenic mice fed with CP-31398. These results suggest that p53 modulating agents can serve as potential chemopreventive for bladder TCC. (Supported in part by NCI-CN53300) Citation Format: Venkateshwar Madka, Yuting Zhang, Qian Li, Altaf Mohammed, Stan Lightfoot, Wu Re Xue, Levy Kopelovich, Chinthalapally V. Rao. p53 stabilizing agent CP-31398 prevents urinary bladder tumor growth and invasion in transgenic UPII-SV40T mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3690. doi:10.1158/1538-7445.AM2013-3690

Abstract 310: Role of AMPKalpha isoforms in bladder tumorigenesis.

Abstract Bladder cancer is the fifth most diagnosed cancer and currently is one of the most expensive cancers to treat due to lifelong surveillance. Bladder cancer is marked by two distinct pathways that result in either noninvasive low grade or invasive high grade cancer. Although low and high grade bladder cancers are defined by driver mutations, similarities such as mTORC1 pathway activation exist. Along these lines rapamycin treatment, an mTORC1 inhibitor, has been shown to suppress tumorigenesis in a mouse model of bladder cancer. These data suggest that mTORC1 activation may be an early tumorigenic event in the bladder. In order to elucidate the mechanism by which the mTORC1 signaling is increased, AMPK (AMP-activated protein kinase), a critical negative regulator of the pathway, was further investigated. AMPK governs a cell's response to metabolic stressors by suppressing anabolic pathways and inhibiting cell growth. Furthermore, it has been observed that AMPK activation is diminished in primary breast cancer. In order to determine whether AMPK activation and/or levels are altered in bladder cancer, a pilot cohort of human samples was obtained and the expression of both AMPKa catalytic isoforms was assessed. Both AMPKa1 and AMPKa2 were significantly suppressed in bladder cancer. AMPKa isoform levels in adjacent non-tumor bladder epithelium and bladder cancer were evaluated in matched patient samples and this revealed cancer-specific AMPKa suppression does indeed exist. Additionally, AMPKa1 protein levels were partially suppressed in low grade cancer and further suppressed in high grade cancer. In contrast, AMPKa2 levels were uniformly suppressed in both high and low grade of cancer. Interestingly, it was observed that AMPKa1 is localized predominantly to the cytoplasm while AMPKa2 was present in cytoplasmic and nuclear compartments. This data suggests that the AMPKa isoforms may have distinct roles in the bladder. To test whether loss of AMPKa2 function promoted bladder tumorigenesis, wild-type and AMPKa2-/- mice received 0.05% BBN [n-butyl-n-(4 hydroxybutyl)] nitrosamine, a rodent bladder carcinogen, in drinking water over the course of 20 weeks. Significant tumors were observed at 20 weeks in both wild-type and AMPKa2-/- mice and bladder weights were significantly higher in AMPKa2-/- mice correlating with larger bladder tumor mass. Additionally, there was a significant increase in the amount of proliferating tumor cells as assessed by Ki67 in the AMPKa2-/- mice. This study suggests that AMPKa isoforms are suppressed in bladder cancer and that AMPKa may play a critical role in bladder cancer progression by loss of expression and tumor suppressive function. Citation Format: Stavros Kopsiaftis, Kathryn N. Phoenix, Katie L. Sullivan, Poornima Hegde, John A. Taylor, Kevin P. Claffey. Role of AMPKalpha isoforms in bladder tumorigenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 310. doi:10.1158/1538-7445.AM2013-310

Abstract 3709: Kawain, a kavalactone, inhibits urinary bladder carcinogenesis in the UPII-mutant Ha-ras transgenic model and associated with inhibition of the mTOR pathway.

Abstract Nonmuscle invasive bladder cancer (NIBC) is the most expensive cancer to treat because of its high rate of recurrence. Therefore, there is an urgent need to develop novel agents that can prevent NIBC recurrence and progression. Kava is a traditional drink of the Pacific Islands for thousands of years and kava drink was linked to unusually low incidences of several cancers, including lung and bladder cancer, despite a high portion of smokers in these populations. Kavalactones, including kawain, are major bioactive components in the kava extract. Given genetic alterations leading to over activity of H-Ras are predominantly found in NIBC, we examined the effect of kawain on the growth of bladder tumors and the survival of the mice in the UPII-mutant Ha-ras transgenic mouse bladder model. This model mimics human NIBC and exhibits enhanced mTOR activity in the bladder tumors. We also investigated the effect of kawain on the mTOR pathway in bladder cancer cell lines. Homozygous mutant Ha-ras transgenic mice were identified through genotyping using Southern blotting analysis. The mice were fed orally with vehicle control or 6 g kawain /kg food for about 5 months starting at 6 weeks of age. More than 68% control food- fed male mice died of ureter obstruction and hydronephrosis within 6 months of age, while more than 78% of mice fed with kawain food survived beyond 6 months of age (P=0.0082, log-rank test). The mean bladder weight of male transgenic mice in kawain treatment group decreased by about 56% compared to those fed with vehicle control diet (mean bladder weights for vehicle control versus kawain treatments are 0. 32±0.13 versus 0.14±0.01, P=0.035). In addition, kawain treatment significantly reduced the incidences of hydronephrosis and hematuria. Mouse pathology analysis revealed that vehicle control treated mice exhibited poorly differentiated transitional cell carcinoma in the bladder, while that kawain treated mice presented well differentiated tumors in the bladder. The bladder tissues from kawain treated mice showed a decreased number of Ki67 positive cells compared to those in vehicle control treated mice, which suggested that kawain inhibited cell proliferation in vivo in bladder tumors. In addition, kawain inhibited the growth of bladder cancer cell lines T24 and UMUC3 with a marked suppression of 4E-BP1 expression and induction of autophagy. The growth inhibitory effect of kawain in mouse embryonic fibroblasts (MEFs) is at least in part dependent on the existence of TSC1. This result suggested that kawain inhibits the mTOR pathway. Therefore, kawain, as a naturally-occurring mTOR inhibitor, deserves further investigation for its potential in preventing the recurrence of NIBC. Citation Format: Zhongbo Liu, Xiaolin Zi. Kawain, a kavalactone, inhibits urinary bladder carcinogenesis in the UPII-mutant Ha-ras transgenic model and associated with inhibition of the mTOR pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3709. doi:10.1158/1538-7445.AM2013-3709

Current State of the Art in Non-Invasive Urodynamics

Lower urinary tract symptoms (LUTS) are highly prevalent and bothersome. The pressure flow or voiding part of urodynamics gives information regarding detrusor contractility and whether bladder outlet obstruction (BOO) is present. However, urodynamics are invasive and can be associated with infection and take time to perform. Moreover it is not clear whether urodynamics are beneficial in assessing LUTS routinely or whether they are able to predict outcomes following surgery for obstruction. As a result several non-invasive techniques and investigations have been developed to try to aid in the work up and management of patients with LUTS / BPE. A PubMed search was performed to review the literature on non-invasive urodynamic tests for voiding dysfunction. Techniques include transition zone index, intravesical prostatic protrusion, prostate circle area ratio, prostatic urethral angle, resistive index, bladder wall and detrusor wall thickness, ultrasound estimated bladder weight, near infrared spectroscopy, condom catheter and penile cuff compression techniques and are summarized in this review.

542 CHARACTERIZATION OF LUMBOSACRAL NEURAL ORGANIZATION OF MICTURITION REFLEX PATHWAYS WITH BLADDER DYSFUNCTION AFTER COMPLETE THORACIC SPINAL CORD TRANSECTION IN RATS

You have accessJournal of UrologyBladder and Urethra: Anatomy, Physiology and Pharmacology (II)1 Apr 2013542 CHARACTERIZATION OF LUMBOSACRAL NEURAL ORGANIZATION OF MICTURITION REFLEX PATHWAYS WITH BLADDER DYSFUNCTION AFTER COMPLETE THORACIC SPINAL CORD TRANSECTION IN RATS Hai-Hong Jiang, Fu-Chen Yang, Brian Balog, Kevin Li, Ching-Yi Lin, Margot Damaser, and Yu-Shang Lee Hai-Hong JiangHai-Hong Jiang Cleveland, OH More articles by this author , Fu-Chen YangFu-Chen Yang Cleveland, OH More articles by this author , Brian BalogBrian Balog Cleveland, OH More articles by this author , Kevin LiKevin Li Cleveland, OH More articles by this author , Ching-Yi LinChing-Yi Lin Cleveland, OH More articles by this author , Margot DamaserMargot Damaser Cleveland, OH More articles by this author , and Yu-Shang LeeYu-Shang Lee Cleveland, OH More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1937AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bladder remodeling after spinal cord injury (SCI) causes related morphologic and functional changes, known as neurogenic bladder. Sacral nervous plasticity also occurs after SCI. These neuroplastic processes may relate to remodeling of the end organs of the bladder and the urethra; however the exact changes and patterns of neuroplasticity after SCI remain unknown. This study aims to identify micturition reflex network at lumbosacral and bladder dysfunction after T8 complete SCI using pseudoabies virus (PRV) transneuronal tracing technology. METHODS Eighty-five adult Sprague-Dawley female rats (225-275g) were used. All rats were divided into 3 groups: naive control, Tx-15 and Tx-45 (15 days and 45 days survival period after complete SCI procedure at the level of T8, respectively). Each group was further subdivided into 2 groups according to PRV injection to bladder body or external urethral sphincter (EUS). Three days after PRV injection to bladder or EUS, animals underwent perfusion followed by tissue harvest of bladder, urethra, and spinal cord. Thirty μm transverse sections were collected from the middle of L4 to the middle of S3 spinal segment for each sample. Animals in Tx-45 group also underwent urodynamic study 3 days before PRV injection. RESULTS Bladder remodeling occurs and becomes exacerbated 15 and 45 days after SCI according to the statistically significant changes of bladder weight. Bladder weight mirrors the bladder dysfunction. The main PRV-positive spinal segment from bladder injection of PRV after SCI moves distal with time from L6-S1 to S1-S2. The number of PRV-labeled neurons was positively correlated with bladder weight 15 days after bladder injection and 45 days after EUS injection, suggesting neuroplasticity after SCI occurs earlier in the pathway involving the bladder than in the pathway involving the urethra. CONCLUSIONS The window of time between bladder and urethra neuroplasticity after T8 complete SCI may indicate the time for full completion of micturition control pathway reorganization at the lumbosacral level. Further studies using specific synaptic stains in the spinal cord will be used to determine the synaptic plasticity and reorganization of the micturition reflex pathway after lumbosacral level SCI. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e222-e223 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hai-Hong Jiang Cleveland, OH More articles by this author Fu-Chen Yang Cleveland, OH More articles by this author Brian Balog Cleveland, OH More articles by this author Kevin Li Cleveland, OH More articles by this author Ching-Yi Lin Cleveland, OH More articles by this author Margot Damaser Cleveland, OH More articles by this author Yu-Shang Lee Cleveland, OH More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

120 CHOLINERGIC MEDIATED DETRUSOR DYSFUNCTION IN A FEMALE RAT MODEL OF PELVIC NEUROPRAXIA

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research (II)1 Apr 2013120 CHOLINERGIC MEDIATED DETRUSOR DYSFUNCTION IN A FEMALE RAT MODEL OF PELVIC NEUROPRAXIA Johanna L. Hannan, Vinson M. Wang, Xiaopu Liu, Fabio Castiglione, Arthur L. Burnett, Petter Hedlund, and Trinity J. Bivalacqua Johanna L. HannanJohanna L. Hannan Baltimore, MD More articles by this author , Vinson M. WangVinson M. Wang Baltimore, MD More articles by this author , Xiaopu LiuXiaopu Liu Baltimore, MD More articles by this author , Fabio CastiglioneFabio Castiglione Milan, Italy More articles by this author , Arthur L. BurnettArthur L. Burnett Baltimore, MD More articles by this author , Petter HedlundPetter Hedlund Lund, Sweden More articles by this author , and Trinity J. BivalacquaTrinity J. Bivalacqua Baltimore, MD More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1499AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Urogynaecological surgeries for cervical or colorectal cancer often result in damage to the neurovascular bundle and pelvic plexus leading to voiding dysfunction. Although surgical techniques have improved mortality rates in women, the prevalence of voiding dysfunctions severely impacts their quality of life. The objective of this study is to characterize the physiological and molecular changes in the bladder that take place following bilateral pelvic nerve injury (BPNI) in female rats. METHODS Female Sprague-Dawley rats (12 wks, n=40) were divided into sham or BPNI in which nerves from the pelvic plexus innervating bladder were bilaterally crushed. Animals were examined 7, 14, 30 and 60 days following BPNI. The bladder was excised and weighed. Detrusor strips were mounted in a myograph and contractile responses to increasing concentrations of carbachol or frequencies of electrical field stimulation (EFS) were assessed. Gene expression of muscarinic type 2 (MR2) and 3 (MR3) receptors was also assessed by RTPCR. RESULTS There was a significant increase in the bladder weight at 7 and 14 days following nerve injury (Sham: 3.1%; 7d: 3.9%; 14d: 4.3% of body weight, p<0.05). Maximal carbachol-mediated contraction was decreased following nerve injury at all time points (Sham: 17.9mN; 7d: 12.3mN; 14d: 12.8mN; 30d: 13.4mN; 60d: 14.8mN; p<0.05). Furthermore, the EC50 was markedly increased at 14 and 30 days after BPNI (Sham: 7.0 pD2; 14d: 6.0 pD2; 30d: 6.2 pD2, p<0.05) indicating that a higher concentration of carbachol was required to achieve 50% of maximal contraction. By 60 days following BPNI, maximal contraction and EC50 to carbachol had returned to sham levels. EFS mediated contraction was also significantly reduced after nerve injury when stimulate at lower frequencies (<8Hz) but all groups achieved the same maximal contraction at high frequency stimulation (32Hz). Gene expression of MR2 was significantly lower 7 days following nerve injury (Sham: 0.91; 7d: 0.34 MR2 relative expression, p<0.05); however gene expression of MR3 was unchanged 7days following BPNI. CONCLUSIONS These results demonstrate that pelvic nerve injury leads to a temporal increase in bladder weight and decreased cholinergic mediated contractions and MR2 gene expression. These data suggest that this animal model can be used to study the mechanism of neuropraxia-induced voiding dysfunction in female rats. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e48 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Johanna L. Hannan Baltimore, MD More articles by this author Vinson M. Wang Baltimore, MD More articles by this author Xiaopu Liu Baltimore, MD More articles by this author Fabio Castiglione Milan, Italy More articles by this author Arthur L. Burnett Baltimore, MD More articles by this author Petter Hedlund Lund, Sweden More articles by this author Trinity J. Bivalacqua Baltimore, MD More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

113 TADALAFIL IMPROVES HYPOXIA, STRUCTURAL DETRUSOR CHANGES AND BLADDER COMPLIANCE AFTER CAVERNOUS NERVE INJURY

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research (II)1 Apr 2013113 TADALAFIL IMPROVES HYPOXIA, STRUCTURAL DETRUSOR CHANGES AND BLADDER COMPLIANCE AFTER CAVERNOUS NERVE INJURY Giorgio Gandaglia, Maarten Albersen, Roberta Buono, Alessandro Nini, Fabio Castiglione, Giorgia Colciago, Fabio Benigni, Alberto Briganti, Francesco Montorsi, and Petter Hedlund Giorgio GandagliaGiorgio Gandaglia Milan, Italy More articles by this author , Maarten AlbersenMaarten Albersen Leuven, Belgium More articles by this author , Roberta BuonoRoberta Buono Milan, Italy More articles by this author , Alessandro NiniAlessandro Nini Milan, Italy More articles by this author , Fabio CastiglioneFabio Castiglione Milan, Italy More articles by this author , Giorgia ColciagoGiorgia Colciago Milan, Italy More articles by this author , Fabio BenigniFabio Benigni Milan, Italy More articles by this author , Alberto BrigantiAlberto Briganti Milan, Italy More articles by this author , Francesco MontorsiFrancesco Montorsi Milan, Italy More articles by this author , and Petter HedlundPetter Hedlund Milan, Italy More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1492AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Erectile dysfunction (ED) and lower urinary tract dysfunction are common after radical prostatectomy (RP). Phosphodiesterase 5 (PDE5) inhibitors are used for post-RP ED and may improve continence recovery after nerve-sparing RP. Information is scarce on detrusor integrity or bladder overactivity (BO) in NS-RP. We therefore studied the effect of tadalafil treatment on bladder function and morphology in rats with cavernous nerve-injury (CNI). METHODS After ethical approval, 21 Sprague Dawley rats underwent CNI. Two-weeks treatment with tadalafil (n=7) was started 2 days prior to CNI. After treatment, cystometries were performed in awake rats. Vehicle-treated (n=7) and sham-operated rats (n=5) were controls. Bladder tissues were harvested for organbath studies and immunofluorescence. Values are mean±SEM. RESULTS Body and bladder weights were similar in all rats. Sham rats had regular micturitions and no non-voiding contractions (NVC). 4 out of 7 (p<0.05) vehicle-treated rats and 2 out of 7 tadalafilü-treated rats had NVC. 2 out of 7 vehicleü-treated rats had incontinence episodes but sham or tadalafilü-treated rats did not. No differences in basal and threshold pressures, micturition intervals or volumes, or bladder capacity were recorded between groups. Micturition pressure amounted to 50±10, 45 ±18, and 25±5 cmH2O for sham, vehicle, and tadalafil (p<0.05). Tadalafil-treated rats had no residual volume (RV) whereas sham and vehicle-treated rats had RV of 0.002 ±0.0008 and 0.03 ±0.005ml. Compliance was 0.06 ±0.01, 0.04 ±0.01 (p<0.05), and 0.1 ±0.02 ml/cmH2O for sham, vehicle-, and tadalafil-treated rats. Nerve-mediated contractions amounted to 48 ±18 (sham), 29 ±6 (tadalafil) and 130±38 % of K+ (vehicle; p<0.05). Emax to carbachol amounted to 96 ±18 (sham), 125 ±25 (tadalafil), and 244 ±69 % of K+ (vehicle; p<0.05). Hypoxia was noted in the suburothelium with hydroxyprobe intenstities of 0.00016 ±0.00003 (sham), 0.00065 ±0.00003 (vehicle; p<0.05), and 0.00039 ±0.00006 (tadalafil). The actin/collagen 3 ratio was 0.4 ±0.08 (sham), 1.53 ±0.4 (vehicle; p<0.05), and 0.8 ±0.1 (tadalafil). Compared to vehicle, detrusor VAChT and CGRP nerves were reduced in CNI rats but were more frequent in tadalafil-treated rats compared to vehicle. CONCLUSIONS CNI induced a BO with NVC without increased frequency. Detrusor from CNI rats had muscarinic supersensitivity, hypoxia and altered actin/collagen ratio. These changes were partially counteracted by 2-weeks daily treatment with tadalafil. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e45-e46 Peer Review Report Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Giorgio Gandaglia Milan, Italy More articles by this author Maarten Albersen Leuven, Belgium More articles by this author Roberta Buono Milan, Italy More articles by this author Alessandro Nini Milan, Italy More articles by this author Fabio Castiglione Milan, Italy More articles by this author Giorgia Colciago Milan, Italy More articles by this author Fabio Benigni Milan, Italy More articles by this author Alberto Briganti Milan, Italy More articles by this author Francesco Montorsi Milan, Italy More articles by this author Petter Hedlund Milan, Italy More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...