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INTRODUCTION AND OBJECTIVE:Although aging or severe bladder ischemia could be involved in progression of detrusor underactivity (DU), the pathophysiological mechanism of DU is not well clarified. H...

INTRODUCTION AND OBJECTIVE:In the lower urinary tract, nitric oxide (NO) is well known as a relaxation factor in the urethra. On the other hand, in the central nervous system, NO is reported to hav...

INTRODUCTION AND OBJECTIVES:Psychological stress exacerbates bladder dysfunction such as overactive bladder (OAB) and bladder pain syndrome/interstitial cystitis (BPS/IC). We previously reported that bombesin (BB), a stress-related neuropeptide, centrally induces facilitation of the rat micturition reflex. Brain BB is reported to activate hypothalamo-pituitary-adrenal axis and sympathetic nerves, as representative responses to stressful conditions, via brain corticotropin-releasing factor (CRF), another stress-related neuropeptide. Therefore, we examined whether brain CRF is involved in the BB-induced frequent urination in rats.METHODS:(1) In urethane anesthetized (0.8 g/kg, ip) male Sprague-Dawley (SD) rats (300-400 g), a catheter was inserted into the bladder to perform cystometry (CMG, 12 ml/h saline infusion). Astressin (a non-selective CRF receptor antagonist, 1 or 3 nmol/rat) was intracerebroventricularly (icv) pretreated 60 min before BB administration (0.03 nmol/rat, icv). CMG was started 60 min b...

INTRODUCTION AND OBJECTIVES:Psychological stress exacerbates symptoms of bladder dysfunction. We previously reported that epibatidine (EP), a nicotinic acetylcholine receptor (nAChR) agonist, centr...

INTRODUCTION AND OBJECTIVES:Psychological stress is involved in the development of the lower urinary tract symptoms. Our previous reports showed that centrally administered a stress-related neurope...

INTRODUCTION AND OBJECTIVES:Ghrelin, a 28 amino acid growth hormone, is widely distributed throughout the peripheral and central nervous systems. Ghrelin′s many physiological functions include grow...

Objective— Nitric oxide (NO) is an important modulator of cardiovascular function. In this study, we examined whether cytosolic phospholipase A 2 α (cPLA 2 α), an initial enzyme in the arachidonic acid pathway, is involved in blood pressure (BP) elevation in a murine model of chronic NO inhibition. Methods and Results— cPLA 2 α gene–deficient mice (cPLA 2 α−/−) and wild-type mice (WT) were administered the NO synthesis inhibitor N ω -nitro- l -arginine methyl ester ( l -NAME) for 4 weeks. Before treatment, BP was comparable in both groups; it increased significantly in the WT but not in the cPLA 2 α−/− after treatment. Bone marrow transplantation experiments showed that cPLA 2 α in blood cells and plasma eicosanoid concentrations were not involved in BP elevation by l -NAME treatment. Activation of cPLA 2 α and subsequent production of eicosanoids in the aortic endothelium but not in aortic smooth muscle cell, heart, or kidney was observed after l -NAME treatment. Aortic ring assays revealed that endothelial function was comparable in both groups of mice before treatment. l -NAME treatment disturbed endothelial function in WT but not in cPLA 2 α−/−. Conclusion— These results suggest that endothelial cPLA 2 α may play a principal role in l -NAME-induced hypertension and may be a target molecule for maintaining endothelial function under NO inhibition.

METHODS: We designed a whole-exome sequencing (WES)based approach using NimbleGen SeqCap EZ Exome V3 sample preparation kit and the Illumina HiSeq 2000 next-generation sequencing platform. We sequenced the exomes of two affected siblings. Exome analysis resulted in the identification of 442 variants in the index patients. All variants that passed filter criteria were validated with Sanger sequencing to confirm familial segregation and absence in the control population. RESULTS: A novel non-synonymous mutation in neuronal PAS 2 domain (NPAS2) was identified in a consanguineous family from Turkey. This mutation in exon 14 (chr2: 101592000 C>G) of NPAS2 segregates with the disease. Family segregation of the variants showed the presence of homozygous mutation in the three brothers with NOA and heterozygous mutation in one brother, and one sister who were fertile. The mutation is not found in the single nucleotide polymorphism (SNP) database, the 1000 Genomes Project, Baylor College of Medicine cohort of 500 Turkish patients (not a founder mutation) or matching 50 fertile controls. The absence of homozygous mutations in NPAS2 in these databases suggests that such gene perturbations are not consistent with fertility, and therefore the observed NPAS2 mutation is considered likely to be pathogenic. CONCLUSIONS: UsingWES, we identified a novel homozygous mutation in NPAS2 as a likely disease-causing variant in three brothers with NOA. Our data reinforce the clinical role of WES in the molecular diagnosis of highly heterogeneous genetic diseases where conventional geneticapproacheshavepreviously failed inachievingaproperdiagnosis.

The current study was done to elucidate the in vivo mechanism of action of intravesical instillation of oxybutynin to treat overactive bladder.In rats receiving oral and intravesical oxybutynin we measured muscarinic receptors in the bladder and other tissues by radioligand binding assay using [3H]NMS ([N-methyl-3H] scopolamine methyl chloride) with the simultaneous measurement of plasma concentrations of oxybutynin and its active metabolite N-desethyl-oxybutynin. Pilocarpine induced salivary secretion was also measured.Following oral administration of oxybutynin there was a significant increase in the apparent dissociation constant (Kd) for specific [3H]NMS binding in the bladder, submaxillary gland, heart and colon of rats at 1 and 3 hours with a consistent decrease in the maximal number of binding sites (Bmax) in the submaxillary gland. Furthermore, a marked and prolonged decrease in pilocarpine induced salivary secretion in rats was observed by oral oxybutynin. In contrast, intravesical instillation of oxybutynin produced a significant increase in Kd for specific [3H]NMS binding in the bladder of rats at 0.5 to 4 hours later and also in the submaxillary gland only at 0.5 hours later. The enhancement in Kd was much larger and longer lasting in the bladder than in the submaxillary gland. Moreover, intravesical oxybutynin had little muscarinic receptor binding activity in the heart and colon, and little significant suppression of pilocarpine induced salivation in rats. The plasma concentrations of oxybutynin and N-desethyl-oxybutynin were much higher in rats receiving oxybutynin orally than intravesically.Intravesical oxybutynin in rats may cause selective binding of bladder muscarinic receptors via a direct local effect, while oral oxybutynin may exert predominant binding of salivary gland receptors.

INTRODUCTION AND OBJECTIVES: In this study, we focused on whether olmesartan, an angiotensin II receptor blocker, or nifedipine, an L-type calcium channel blocker, could prevent dysfunction of NO-induced relaxation in corpus cavernosum smooth muscle in the spontaneously hypertensive rat (SHR). METHODS: Twelve-week-old male SHRs, and Wistar rats as aged-matched normotensive controls were used in this study. The treatment groups were administered olmesartan (1 or 3mg/kg, per orally (p.o.); Olm1 or Olm3, respectively) or nifedipine (30mg/kg, p.o.; Nif) for six weeks daily. Penile function was evaluated by organ bath studies with norepinephrine-induced contractions and acetylcholineinduced relaxations. Penile cGMP concentrations and mRNA levels of eNOS and nNOS were investigated. RESULTS: The SHR showed significantly increased blood pressure (BP), norepinephrine-induced hyper-contractions, acetylcholine-induced hypo-relaxations, decreased cGMP concentrations, and decreased eNOS and nNOS mRNA levels in the penile tissue. Treatment with high dose of olmesartan significantly decreased BP, significantly increased the penile cGMP concentrations, partially normalized the eNOS and nNOS mRNA levels, significantly decreased the norepinephrine-induced contractions, and normalized the acetylcholineinduced relaxation compared to the SHR. Nifedipine significantly decreased BP, increased cGMP, and normalized the hyper-contractions and hypo-relaxations observed in the SHR group, while the mRNA levels of eNOS and nNOS were significantly lower compared to the Wistar. CONCLUSIONS: Our results indicate that the dysfunction in the SHR penile tissue could be better reversed by olmesartan treatment and to a slightly lesser extent by nifedipine treatment.

INTRODUCTION AND OBJECTIVES: Vaginal delivery may cause trauma to pelvic floor muscle (PFM). Several authors attribute urogynecological symptoms that appear during the climacteric period to PFM dysfunction secondary to pregnancy and delivery. This neuromuscular injury may cause weakness in pelvic support structures. The aim of this study was to evaluate the impact of parity and mode of delivery on PFM strength in healthy continent women. METHODS: Hundred forty healthy continents women were divided into 4 groups according to age: G1 (n 1⁄4 34) 30-40 years old; G2 (n 1⁄4 38) 41-50 years old; G3 (n 1⁄4 35) 51-60 years old and G4 (n 1⁄4 33) over 60 years old. The following parameters were analyzed: obstetric and gynecological data assessed by clinical questionnaire; body mass index (BMI); subjective evaluation of PFM strength assessed by transvaginal digital palpation (TDP) and objective evaluation of PFM using perineometer. RESULTS: BMI was significantly higher in G4 compared to G1 group (Table 1). The number of pregnancies was significantly higher in G4 compared to G2 (Table 1). The number of vaginal deliveries was significantly higher in G4 compared to G1 and G3 (Table1). There was a positive linear relationship between age and BMI (r 1⁄4 0,215, p 1⁄4 0,011). There was no statistical correlation among different groups in PFM strength assessed by TDP and perineometer. There was not linear relationship between PFM strength and number of pregnancies (r1⁄4 0.70 p1⁄4 0.409) and vaginal deliveries (r1⁄4 -0.026 p1⁄40.758). CONCLUSIONS: These data demonstrate that despite the increase in BMI, number of pregnancies and vaginal deliveries with aging, these parameters did not impact on PFM strength in continent women.

of age-related bladder dysfunction has not been fully clarified yet. In the present study, we aimed to investigate age-related changes in the bladder function of the mouse by using precise frequency volume (FV) measurements, in vitro contractile studies and histological examination of detrusor strips. Gender differences in the age-related changes were also determined. METHODS: C57/BL6 male and female mice were divided into two groups: control (CONT: 12 months-old; n¼8 of each sex) and aged (AGED: 27-30 months-old; male: n¼6, female: n¼7). After FV measurements, in vitro organ bath studies using detrusor strips were carried out to evaluate their contractile responses to high K+ (KCl: 62 mM), carbachol (CCh: 10-3-10-8 M), and to electrical field stimulation (EFS; 2-20 Hz) in the absence and presence of atropine (10-6 M), a,b-methylene ATP (M-ATP: 10-5 M x 5 times). In separate detrusor strips, the muscle/collagen ratio was evaluated by using Masson-trichrome staining. RESULTS: In the FV measurement, there were no significant differences in mean voided volume per micturition between CONT and AGED in either sex, although AGED of both sexes had larger total voided volume per day. Female AGED showed a decreased mean uroflow rate. The contractile responses to high K+ were weaker in AGED of both sexes (Fig.). In addition, those to CCh and EFS were decreased with aging in both sexes, but when standardized by the high K+-induced contraction, the contractile responses to CCh were stronger in AGED of both sexes (Fig.), and those to EFS were stronger only in female AGED. Moreover, AGED of both sexes showed a decreased atropine-sensitive component and an increased M-ATP-sensitive component of the EFS-induced contractions. The muscle/collagen ratio in the muscle layer did not change significantly with aging in either gender. CONCLUSIONS: The current in vitro functional studies with analysis of muscle/collagen ratio indicate the intrinsic contractile property of detrusor smooth muscle in both sexes is potentially impaired with aging. The findings that the contractile responses standardized by the high K+ -induced contraction to CCh and EFS were increased in the aged bladder, suggest a compensatory mechanism for the myogenic impairment. This compensatory mechanism may make the impairment of in vivo bladder function minimal. Source of Funding: none

As there are significant amounts of endothelin (ET) receptors in the mammalian urinary tract, we investigated the pharmacological properties and localization of ET receptors in the rabbit lower urinary tract as a function of age.The characteristics of ET receptors in bladder dome, trigone and urethra of 6 weeks and 6 months old male rabbits were determined using muscle bath and autoradiographic techniques.ET-1 produces significant contractile responses in smooth muscle strips from bladder dome, trigone, and urethra in both 6 weeks and 6 months old rabbits. Although there was no significant difference in the maximum contractile response of urethral muscle strips to ET-1 between 6 weeks and 6 months old rabbits, the maximum responses to ET-1 were higher in both bladder dome and trigone of 6 weeks than 6 months old rabbits. A selective ETA receptor antagonist, BQ 610, shifted the concentration response curve to ET-1 to the right without decreasing maximal contractile responses in all regions from both age groups, whereas a selective ETB receptor antagonist, IRL 1038, had no significant effect on the contractile response in these tissues. Autoradiographic studies indicate that both ET receptor subtypes are expressed in bladder dome, trigone, and urethra with the ETA subtype being located only in the smooth muscle layers and the ETB subtype being located in both the urothelial and smooth muscle layers.Our data indicate the presence of region- and age-dependent differences in the contractile properties of ET receptors in the male rabbit lower urinary tract. Although both ETA and ETB receptor subtypes are present in the smooth muscle layers, the ETA receptor is the sub-type that is primarily involved in the mediation of contractions.

Purpose: The subtype specificity, localization and distribution of urethral alpha1-adrenoceptors were studied in the male rabbit urethra.Materials and Methods: The properties of the urethral alpha1-adrenoceptors were investigated using radioligand receptor binding and light microscopic autoradiography with [(125) I]iodo-2-[b-(4-hydroxyphenyl)-ethylaminomethyl]tetralone (HEAT), and immunohistochemistry with monoclonal anti-alpha smooth muscle actin and anti-alpha sarcomeric actin antibodies.Results: Saturation experiments with [(125) I]HEAT demonstrated the presence of significant amounts of a single high affinity binding site for alpha1 adrenoceptors in the male rabbit urethra. The pharmacological profile of the alpha1 adrenoceptors in rabbit urethra, determined by inhibition experiments with subtype selective alpha1 adrenoceptor antagonists, was characterized by the following rank order of potency of inhibition constants (Ki values): prazosin

To elucidate the subtype specificity of muscarinic cholinergic receptors in mediating contractile responses in the lower urinary tract, we investigated contractile and biochemical properties of muscarinic receptors in bladder dome, bladder base and urethra of the rabbit. Isometric contractile response curves to increasing concentrations of carbachol were constructed in the absence and presence of various concentrations of subtype selective muscarinic antagonists. Bladder dome, bladder base, and urethra demonstrate different characteristics in terms of efficacy and potency with respect to carbachol-induced contractile responses. Emax values are significantly larger and ED50 values are significantly smaller in bladder dome and bladder base than in urethra. Calculation of the pA2 values, the negative logarithm of the antagonist affinity constant (KB), for a series of muscarinic antagonists, i.e., atropine (nonselective), pirenzepine (M1 selective), methoctramine (M2 selective), and 4-DAMP (M1/M3 selective) indicate that the carbachol-induced contractile response in bladder dome and bladder base is mediated through the M3 receptor subtype whereas the carbachol-induced contractile response in urethra is probably mediated through the M1 and/or M3 and possibly M2 subtypes. Muscarinic cholinergic antagonists inhibit [3H]quinulidinyl benzilate binding to bladder dome, bladder base and urethra with the following rank order of affinities: atropine4-DAMPmethoctraminepirenzepine. The binding data indicate the predominance of the M2 receptor subtype in all three regions.

sepsis by obstructive acute pyelonephritis were admitted to our department and received a total of 54 emergency drainages of upper urinary tract for urinary drainage. We evaluated the characteristics of these patients and events retrospectively to determine risk factors for septic shock using univariate (chi-square test) and multivariate analysis (logistic regression analysis). RESULTS: Among 54 events of sepsis, we identified 34 events of septic shock requiring vasopressor therapy. One patient (1.8%) died of septic shock. Patients who developed septic shock were received ureteral drainage earlier from the onset of symptoms than those without septic shock (1.20 vs 4.44 days, p 0.001, t-test).Using univariate analysis, pyelonephritis caused by ureteral calculi, transient thrombocytopenia ( 120,000/mm3), coagulopathy (prothrombin time International normalized ratio (PT-INR) 1.20), leukocytopenia ( 4,000/mm3) before ureteral drainage and positive blood culture were correlated with septic shock (p 0.05). Multivariate logistic regression analysis showed thrombocytopenia before ureteral drainage (OR 24.16; 95% CI, 2.98-195.52; P 0.0028) and positive blood culture (OR 8.10; 95% CI, 1.09-60.33; P 0.0413) were independent risk factors associated with septic shock. CONCLUSIONS: Our study, although it is retrospective, suggests that positive blood culture and thrombocytopenia could predict septic shock. Early and intensive interventions may be recommended to prevent fatal outcomes in patients with these factors.

PC-3, DU-145, LNCaP, and CRW22Rv1 cell lines upon ZA treatment. Formation of the LC3-II protein was detected by Western blotting. LC3-II incorporation into autophagosomes was detected by immunofluorescent (IF) staining. Formation of acidic organelles was detected by acridine orange (AO) staining. Rescue experiments using an apoptosis inhibitor and/or an autophagy inhibitor were performed by MTT assays. RESULTS: ZA exhibited dose and time-dependent growth inhibition on four human prostate cancer cells. Autophagic cell death was detected by the formation of LC3-II protein as early as 24 hours exposure to 100nM of ZA. LC3-II and caspase 3 processing was detected at 6 days post-treatment. Acidic organelles were detectable by AO staining, and IF showed round-up and condensed staining of LC3-II, suggesting the formation of autophagosomes in the cytoplasm during autophagic cell death. The rescue of cell growth occurred only with administration of both an apoptosis and autophagy inhibitor during ZA treatment, indicating that ZA induces prostate cancer death by both apoptotic and autophagic cell death. CONCLUSIONS: This is the first study to show that ZA markedly inhibited human prostate cancer cell growth through autophagic cell death. ZA exhibits its anticancer activity via both apoptosis and autophagy. These findings can potentially contribute to the beneficial use of ZA in prostate cancer treatments.

We investigated bladder function in acute urinary retention and subsequent catheterization in rat bladders.The penile urethra in rats was clamped with a small clip and cystostomy was performed to infuse 3 ml. of saline for inducing acute urinary retention. At 30 minutes after the induction of urinary retention the cystostomy was opened to empty the bladder. In functional studies contractile responses to carbachol were measured in group 1-before, group 2-at 3 ml. of urinary retention, group 3-at 3 ml. of urinary retention exposed for 30 minutes and group 4-30 minutes after catheterization. Moreover, in vivo real-time monitoring of blood flow and vesical pressure were measured in the bladders with a laser Doppler flowmeter and cystometrography, respectively. Malonaldehyde and 4-hydroxyalkenals were measured by colorimetric assay in these groups.In functional studies the mean maximum contractile response value plus or minus standard deviation of carbachol-to-bladder in groups 1 to 4 was 11.8 +/- 1.3, 11.9 +/- 1.7, 9.8 +/- 0.8 and 6.9 +/- 0.7 gm./mm.2, respectively. In real-time monitoring of blood flow and vesical pressure acute urinary retention significantly decreased blood flow and increased vesical pressure, and subsequent catheterization increased blood flow and decreased vesical pressure in the bladders. The concentrations of malonaldehyde and 4-hydroxyalkenals in the bladders in group 4 were significantly higher than in the other groups.Our data indicate that bladder dysfunction after catheterization is partially caused by free radicals, which have an important role in bladder dysfunction during acute urinary retention.

An 85-year-old Japanese man presented with abdominal discomfort and general fatigue. History included (stage A1) prostate cancer, G1T1a transitional cell carcinoma of the bladder for 4 years and ulcerative colitis, which had been treated with 4.5 gm. sulfasalazine daily for 14 years. Laboratory examination showed renal failure (serum blood urea nitrogen was 26 mg./dl. and creatinine was 1.71 mg./dl.). Ultrasonography, computerized tomography and x-ray revealed bilateral hydronephrosis, right multiple renal stones, a ureteropelvic junction stone and a left ureterovesical junction stone (see figure). The left ureteral stone was spontaneously discharged 5 days after the initial consult. The patient underwent 4 sessions of shock wave lithotripsy and percutaneous nephrolithotomy to remove stones in the right renal pelvis and ureteropelvic junction, after ureteral stenting. After stenting, renal function returned to normal (blood urea nitrogen 10 mg./dl. and serum creatinine 0.89 mg./dl.). Stone analysis revealed acetylsulfapyridine, which is a metabolite of sulfasalazine. After the final diagnosis, sulfasalazine was replaced with mesalamine (5-aminosalicylic acid) to treat the ulcerative colitis, and at 6-month followup no recurrence of stone disease was observed.

Purpose: We describe a simple technique to replace an obstructed ureteral stent in patients with an ileal conduit and ureteral stenosis.Materials and Methods: Our study included 4 patients with an ileal conduit and ureteral stenosis. A total of 10 obstructed ureteral stents were replaced with a new stent using this simple technique under radiographic guidance.Results: All obstructed ureteral stents were successfully replaced with a new stent without endoscopy, percutaneous nephrostomy or any special devices.Conclusions: Our technique is simple, easy and highly successful for changing an obstructed ureteral stent.