S-nitrosylation of endothelial nitric oxide synthase impacts erectile function.

Neuronal and endothelial nitric oxide synthases (nNOS and eNOS respectively) play major roles in generating the nitric oxide bioactivity necessary for erectile function. S-nitrosylation has been shown to regulate NOS activity. The presence of S-nitrosylated NOS in the penis and the impact of NOS S-nitrosylation/denitrosylation on erectile function were examined. S-nitrosylated forms of NOS were identified by biotin-switch assay followed by western blot analysis. Erectile function in S-nitrosoglutathione reductase deficient (GSNO+/-) and null (GSNO-/-) mice were assessed by continuous cavernous nerve electrical stimulation (CCNES). Glutathione ethyl ester (GSHee) was used to manipulate S-nitrosylated NOS levels. Immunohistological and immunofluorescence analyses were used to identify the location of eNOS and GSNO-R in corporal tissue. eNOS and nNOS were S-nitrosylated in unstimulated penises of the mice. CCNES resulted in a time-dependent increase in eNOS S-nitrosylation with peak eNOS S-nitrosylation observed during detumescence. S-nitrosylated nNOS levels were unchanged. Intracorporal injection of GSHee reduced S-nitrosylated eNOS levels, enhancing time to maximum intracorporal pressure (ICP). eNOS and GSNO-R co-localize to the endothelium of the corpus cavernosum in the mouse and the human. ICP measurements obtained during CCNES demonstrate GSNO-R+/- and GSNO-R-/- animals cannot maintain an elevated ICP. Results suggest eNOS S-nitrosylation/denitrosylation is an important mechanism regulating eNOS activity during erectile function. GSNO-R is a key enzyme involved in the eNOS denitrosylation. The increase in eNOS S-nitrosylation (inactivation) observed with tumescence may begin a cycle leading to detumescence. Clinically this may indicate that alterations in the balance of S-nitrosylation/denitrosylation either directly or indirectly contribute to erectile dysfunction.

Urinary bladder organ hypertrophy is partially regulated by Akt1-mediated protein synthesis pathway.

AIMS: The present study aims to investigate the role of Akt in the regulation of urinary bladder organ hypertrophy caused by partial bladder outlet obstruction (pBOO). MAIN METHODS: Male rats were surgically induced for pBOO. Real-time PCR and western blot were used to examine the levels of mRNA and protein. A phosphoinositide 3-kinase (PI3K) inhibitor LY294002 was used to inhibit the activity of endogenous Akt. KEY FINDINGS: The urinary bladder developed hypertrophy at 2 weeks of pBOO. The protein but not mRNA levels of type I collagen and α-smooth muscle actin (αSMA) were increased in pBOO bladder when compared to sham control. The phosphorylation (activation) levels of Akt1 (p-Ser473), mammalian target of rapamycin (mTOR), p70S6 kinase (p70S6K), and 4E-BP1 were also increased in pBOO bladder. LY294002 treatment reduced the phosphorylation levels of Akt1 and 4E-BP1, and the protein levels of type I collagen and αSMA in pBOO bladder. The mRNA and protein levels of proliferating cell nuclear antigen (PCNA) were increased in pBOO bladder, and PCNA up-regulation occurred in urothelial not muscular layer. LY294002 treatment had no effect on the mRNA and protein levels of PCNA in pBOO bladder. LY294002 treatment partially reduced the bladder weight caused by pBOO. SIGNIFICANCE: pBOO-induced urinary bladder hypertrophy is attributable to fibrosis, smooth muscle cellular hypertrophy, and urothelium cell hyper-proliferation. Akt1-mediated protein synthesis in pBOO bladder contributes to type I collagen and αSMA but not PCNA up-regulation. Target of Akt1 is necessary but not sufficient in treatment of urinary bladder hypertrophy following pBOO.

A Review on Functional Electrical Stimulation of Bladder and Bowel in Spinal Cord Injury.

The purpose of this collective review is to examine the use of functional electrical stimulation for incontinence. The Finetech-Brindley bladder system enhances voiding through stimulation via electrodes implanted around the ventral sacral roots. Detrusor hyperreflexia is eliminated through complete dorsal rhizotomy, which results in loss of reflex defecation and reflex erection/reflex lubrication. Consequently, a new system is being devised in which functional electrical stimulation for incontinence in spinal cord injury can be achieved without dorsal rhizotomy.

Spontaneous voiding by mice reveals strain-specific lower urinary tract function to be a quantitative genetic trait.

Lower urinary tract (LUT) symptoms become prevalent with aging and affect millions; however, therapy is often ineffective because the etiology is unknown. Existing assays of LUT function in animal models are often invasive; however, a noninvasive assay is required to study symptom progression and determine genetic correlates. Here, we present a spontaneous voiding assay that is simple, reproducible, quantitative, and noninvasive. Young female mice from eight inbred mouse strains (129S1/SvImJ, A/J, C57BL/6J, NOD/ShiLtJ, NZO/H1LtJ, CAST/EiJ, PWK/PhJ, and WSB/EiJ) were tested for urination patterns on filter paper. Repeat testing at different times of the day showed minimal within-individual and within-strain variations, but all parameters (spot number, total volume, percent area in primary void, corner voiding, and center voiding) exhibited significant variations between strains. Calculation of the intraclass correlation coefficient, an estimate of broad-sense heritability, for each time of day and for each voiding parameter revealed highly significant heritability [spot number: 61%, percent urine in primary void: 90%, and total volume: 94% (afternoon data)]. Cystometrograms confirmed strong strain-specific urodynamic characteristics. Behavior-voiding correlation analysis showed no correlation with anxiety phenotypes. Diagnostically, the assay revealed LUT symptoms in several systems, including a demonstration of voiding abnormalities in older C57BL/6J mice (18-24 mo), in a model of protamine sulfate-induced urothelial damage and in a model of sucrose-induced diuresis. This assay may be used to derive pathophysiological LUT readouts from mouse models. Voiding characteristics are heritable traits, opening the way for genetic studies of LUT symptoms using outbred mouse populations.

Lower urinary tract symptom improvement after radical prostatectomy correlates with degree of prostatic inflammation.

OBJECTIVE: To determine if prostatic inflammation at the time of radical prostatectomy (RP) was associated with the International Prostate Symptom Score (IPSS). METHODS: We performed a proof of principle analytic case control study of patients who underwent RP between January 2005 and August 2008 for lower urinary tract symptoms (LUTS). We reviewed pathology slides of those who had a change of 4 points or greater, as measured by the IPSS and correlated inflammation with change in IPSS. Multivariate linear regression analyses were performed to determine the association of IPSS with degree of inflammation based on the number of inflammatory cells. RESULTS: Of 249 patients, 136 had complete data and 47 (18.8%) underwent pathologic review. The median change in IPSS for the study cohort was -7.0 points compared to +1.0 point for the control cohort. On univariate analysis, the average improvement in IPSS in patients with severe inflammation was (r = -6.02, 95% confidence interval [CI] -11.0 to -1.1, P = .018) after RP. On multivariate analysis, adjusting for age, body mass index (BMI), year of surgery, history of prostatitis, Gleason score, prostate-specific antigen (PSA), prostate weight, and nerve sparing status, only patients with severe prostatic inflammation had significant improvement in their IPSS (r = -5.93, 95% CI -10.81 to -1.04, P = .004). CONCLUSION: Prostatic inflammation measured in prostatectomy specimens is associated with worse baseline IPSS than matched cohorts. Specifically, severe inflammation is an independent predictor of IPSS improvement at 1 year after RP.

The association of depression, anxiety and nocturia: a systematic review.

PURPOSE: This systematic review focuses on the relationship between nocturia and depression/anxiety. Our objective is to provide an overview of current data on the epidemiology, pathophysiology and patient management implications of the association between nocturia and depression/anxiety. MATERIALS AND METHODS: We queried PubMed®, Web of Science® and Embase™ in July 2012 to identify abstracts, and original, review and editorial articles on nocturia and mood disorders, specifically depression and anxiety. The search was done using the key words "nocturia," "depression" and "anxiety." We complied with the Assessment of Multiple Systemic Reviews (AMSTAR) instrument. We retrieved a total of 500 records, including 95, 81 and 324 from PubMed, Web of Science and Embase, respectively. RESULTS: Cross-sectional (level 3) data indicated that nocturia and depression/anxiety are strongly associated. One prospective study contended that depression leads to nocturia in a unidirectional relationship. Nocturia poses a greater risk for depression in men vs women. Results conflict on the effect of serotonin reuptake inhibitors on nocturia. CONCLUSIONS: The results of this systematic review suggest a bidirectional association between depression and nocturia. The relationship between anxiety and nocturia is less clear. Practicing clinicians should consider administering a brief self-administered scale to assess for depression in patients with nocturia.

Bladder and urethral function in a mouse model of cavernous nerve injury.

AIMS: To determine whether cavernous nerve injury (CNI) alters lower urinary tract function, we assessed bladder and urethral function over time in a mouse model of CNI. METHODS: Twelve-week-old male C57BL/6 mice were divided into three groups: unoperated (UO; n = 6), sham-operated (SO; n = 18), and bilateral CNI (n = 30) group. At 1, 2, 4, 6, 8, 10 days bladder and urethral function were evaluated in these three groups using cystometry (CMG) and leak point pressure (LPP) recording under anesthesia. RESULTS: There was no significant difference in maximum detrusor pressure between groups at all times. Compared with the UO group, bladder compliance, and capacity in the CNI group were significantly decreased at Days 1, 2, 4 (P < 0.05) and recovered gradually from Day 6 to Day 10. In the SO group, they were decreased at Day 1, however, recovered more rapidly than the CNI group. Non-voiding contractions (NVC) developed in the CNI group at all times. Intercontraction interval were significantly decreased in SO and CNI groups and recovered more rapidly in SO group. In the SO group NVC were observed only at Days 1 and 2. LPP in the CNI group was decreased significantly at Days 1 and 2 (P < 0.05) and rapidly recovered with time compared with the UO and SO groups. CONCLUSION: In a mouse model of CNI, a transient decrease in bladder compliance, capacity, LPP and increased NVC was observed. These changes gradually recovered from Day 6 after CNI. Our findings suggest that CNI may affect bladder and urethral function, but alterations are reversible.

Increased phosphodiesterase type 5 levels in a mouse model of type 2 diabetes mellitus.

INTRODUCTION: Diabetes mellitus (DM) is a major risk factor for developing erectile dysfunction (ED) and men with DM are often less responsive to phosphodiesterase type 5 (PDE5) inhibitors than ED due to other causes. AIMS: The aim of this study was to explore potential mechanisms whereby PDE5 inhibitors may have reduced efficacy in type 2 DM. METHODS: At 4 weeks of age, mice were either fed a high-fat diet (HFD) for 22-36 weeks or fed regular chow (control). An additional group of mice in the same genetic background had a genetic form of type 1 DM. MAIN OUTCOME MEASURES: Glucose tolerance testing, intracorporal pressures (ICPs), oxidative stress (OS), apoptotic cell death (active caspase-3 and apostain), PDE5, p53, and cyclic guanosine monophosphate (cGMP) levels, and histological examination of inflow arteries were performed in mice fed a HFD and control mice. A group of mice with type 1 DM were studied for PDE5 expression levels. RESULTS: All mice fed a HFD had impaired glucose tolerance compared with the age-matched mice fed on standard chow diet (control). HFD fed mice had reduced maximum ICPs following in vivo cavernous nerve electrical stimulation and increased apoptotic cell death, OS, and p53 levels in the corporal tissue. Interestingly, PDE5 levels were increased and cGMP levels were decreased. In contrast, mice with type 1 DM did not have increases in PDE5. CONCLUSIONS: Taken together, our results suggest that type 2 DM-induced ED is associated with findings that could lead to reduced cGMP and may account for reduced efficacy of PDE5 inhibitors.

Establishing the subspecialty of female pelvic medicine and reconstructive surgery in the United States of America.

OBJECTIVE: In this review I describe the history leading to the creation of the subspecialty of female pelvic medicine and reconstructive surgery and its fellowships, the process involved in the current requirements for subspecialty certification and fellowship applications, and the implications for urological training. RESULTS AND CONCLUSIONS: The route to subspecialty certification and fellowships for female urology in the USA is a lesson in politics, education, medical rivalries and perseverance, with the goal of improving care for women. This decade-long journey culminated in the recognition of a separate subspecialty by the American Board of Medical Specialties in 2011, accreditation by the American Council for Graduate Medical Education in 2012, and certification to be awarded by the Boards of Obstetrics and Gynecology and Urology in 2013. It remains to be seen whether this effort will improve resident education and patient care, or represent a marketing tool in the competitive USA healthcare environment. While many of the details and regulatory issues are specific to the USA, elements of the curriculum and procedures should be relevant to other countries.

Associations of commonly used medications with urinary incontinence in a community based sample.

PURPOSE: We examined the association between the use of medications and the prevalence of urinary incontinence in gender specific analyses of a community based, representative sample. MATERIALS AND METHODS: A population based epidemiological study was conducted of 5,503 men and women 30 to 79 years old residing in Boston, Massachusetts (baseline data collected from 2002 to 2005). Urological symptoms were ascertained in a 2-hour, in person interview. Urinary incontinence was defined as urine leakage occurring weekly or more often during the last year. Medications used in the last month were considered current use. Associations of 20+ medications and prevalent urinary incontinence were examined using multivariate logistic regression (ORs and 95% CIs) with adjustments for known urinary incontinence risk factors. RESULTS: The prevalence of urinary incontinence in the analysis sample was 9.0% in women and 4.6% in men. For women the prevalence was highest among users of certain antihistamines (28.4%) and angiotensin II receptor blockers (22.9%). For men the prevalence was highest among angiotensin II receptor blocker (22.2%) and loop diuretic (19.1%) users. After final multivariate adjustment there were significant positive associations for certain antihistamines, beta receptor agonists, angiotensin II receptor blockers and estrogens with urinary incontinence in women (all ORs greater than 1.7), and a borderline significant association for anticonvulsants (OR 1.75; 95% CI 1.00, 3.07). Among men only anticonvulsants were associated with urinary incontinence after final adjustments (OR 2.50; 95% CI 1.24, 5.03), although angiotensin II receptor blockers showed an adjusted association of borderline significance (OR 2.21; 95% CI 0.96, 5.10). CONCLUSIONS: Although a cross-sectional analysis cannot determine causality, our analysis suggests certain medications should be further examined in longitudinal analyses of risk to determine their influence on urological symptoms.

Confocal fluorescence microscopy in a murine model of microdissection testicular sperm extraction to improve sperm retrieval.

PURPOSE: Microdissection testicular sperm extraction markedly improves the sperm retrieval rates in men with nonobstructive azoospermia. However, localizing sperm foci can be time-consuming and it is not always successful. Fiberoptic confocal fluorescent microscopy offers the advantage of rapid in vivo detection of fluorescently labeled sperm in the seminiferous tubules. MATERIALS AND METHODS: After establishing the feasibility of fiberoptic confocal fluorescent microscopy to identify antibody labeled sperm in vivo C57/B6 mice underwent intraperitoneal injection of busulfan to induce azoospermia. During spermatogenesis reestablishment at approximately 16 weeks the mice were anesthetized and the testes were delivered through a low midline incision. Fluorescein isothiocyanate labeled antibody to intra-acrosomal protein Hs-14 was injected retrograde into a single murine rete testis. The testes were imaged in vivo with fiberoptic confocal fluorescent microscopy and sperm foci were detected. The respective seminiferous tubules were excised and squash prepared for immunofluorescence microscopy. RESULTS: Sperm foci were identified in the testis injected with fluorescently tagged antibody by in vivo fiberoptic confocal fluorescence microscopy. The contralateral control testis of each mouse showed no specific signal. Immunofluorescence microscopy of the excised tubules provided morphological confirmation of the presence of labeled sperm with an absence in controls. Findings were consistent in the feasibility portion of the study and in the busulfan model of nonobstructive azoospermia. CONCLUSIONS: Fiberoptic confocal fluorescent microscopy was feasible during microdissection testicular sperm extraction in an azoospermic mouse model to identify fluorescently labeled sperm in vivo. Translation to the clinical setting could decrease operative time and improve the sperm harvest rate.