Effect of estrogen on molecular and functional characteristics of the rodent vaginal muscularis.

INTRODUCTION: Vaginal atrophy is a consequence of menopause; however, little is known concerning the effect of a decrease in systemic estrogen on vaginal smooth muscle structure and function. As the incidence of pelvic floor disorders increases with age, it is important to determine if estrogen regulates the molecular composition and contractility of the vaginal muscularis. AIM: The goal of this study was to determine the effect of estrogen on molecular and functional characteristics of the vaginal muscularis utilizing a rodent model of surgical menopause. METHODS: Three- to 4-month old Sprague-Dawley rats underwent sham laparotomy (Sham, N = 18) or ovariectomy (Ovx, N = 39). Two weeks following surgery, animals received a subcutaneous osmotic pump containing vehicle (Sham, Ovx) or 17ß-estradiol (Ovx). Animals were euthanized 1 week later, and the proximal vagina was collected for analysis of contractile protein expression and in vitro studies of contractility. Measurements were analyzed using a one-way analysis of variance followed by Tukey's post hoc analysis (α = 0.05). MAIN OUTCOME MEASURES: Protein and mRNA transcript expression levels of contractile proteins, in vitro measurements of vaginal contractility. RESULTS: Ovariectomy decreased the expression of carboxyl-terminal myosin heavy chain isoform (SM1) and h-caldesmon and reduced the amplitude of contraction of the vaginal muscularis in response to KCl. Estradiol replacement reversed these changes. No differences were detected in the % vaginal muscularis, mRNA transcript expression of amino-terminal MHC isoforms, l-caldesmon expression, and maximal velocity of shortening. CONCLUSION: Systemic estrogen replacement restores functional and molecular characteristics of the vaginal muscularis of ovariectomized rats. Our results indicate that menopause is associated with changes in the vaginal muscularis, which may contribute to the increased incidence of pelvic floor disorders with age.

Calcineurin mediates bladder wall remodeling secondary to partial outlet obstruction.

We hypothesized that the calcineurin-nuclear factor of activated T-cells (NFAT) pathway is activated following partial bladder outlet obstruction (pBOO), which would allow for pharmacologic treatment to prevent the ensuing bladder wall hypertrophy. Using a model of pBOO in male mice, we were able to demonstrate increased nuclear importation of the transcription factors NFAT and myocyte enhanching factor 2 both of which are under control of calcineurin in both the whole bladder wall as well as the urothelium. We further confirmed that this pathway was activated using transgenic mice containing an NFAT-luciferase reporter construct. Mice were randomized following pBOO to treatment with or without cyclosporine A (CsA), a known inhibitor of calcineurin. The bladder-to-body mass ratio (mg bladder wt/g body wt) of 0.95 ± 0.03 in shams increased to 3.1 ± 0.35 following pBOO, and it dropped back to 1.7 ± 0.22 in the CsA+ group (P < 0.001). Luciferase values (RLU) of 1,130 ± 133 in shams increased to 2,010 ± 474 following pBOO and were suppressed to 562 ± 177 in the CsA+ group (P < 0.05). The myosin heavy chain mRNA (A/B) isoform ratio of 0.07 ± 0.03 in shams increased to 1.04 ± 0.19 following pBOO but it diminished to 0.24 ± 0.1 in the CsA+ group (P < 0.001). In vitro whole organ physiology studies demonstrated improved responses in those bladders from mice treated with CsA. The mRNAs for all four known calcineurin-responsive NFAT isoforms are expressed in the bladder wall, although NFATc(3) and NFATc(4) predominate. Both NFATc3 and NFATc4 are expressed in urothelial as well as smooth muscle cells. We conclude that pBOO activates the calcineurin-NFAT pathway and that CsA treatment decreased bladder hypertrophy, shifted the pattern of myosin isoform mRNA expression back toward that seen in normal controls, and resulted in improved in vitro whole organ performance.

Predictive power of objectivation of phimosis grade on outcomes of topical 0.1% betamethasone treatment of phimosis.

OBJECTIVE: To evaluate the predictive power of the objectivation of the phimosis grade according to the classification defined by Kikiros and Woodward, with regard to the expected efficacy of 0.1% betamethasone cream as a treatment option. METHODS: From October 2010 to May 2011, a total of 55 boys (aged <10 years) were treated for phimosis at our department. An assessment of the category of phimosis and the retractability of the foreskin, according to the classification of Kikiros and Woodward, was performed. The proposed treatment options included complete circumcision or topical treatment with steroid cream (0.1% betamethasone-17-valerate). RESULTS: Of the 55 patients, 19 (34.5%) underwent conventional circumcision, and 36 (65.5%) were treated with an 8-week course of topical steroid cream. The mean age was 3.9 years (range 0.6-10). Grade 1, 2, 3, 4, and 5 phimosis was seen in 1 (2.8%), 4 (11.1%), 8 (22.2%), 16 (44.4%), and 7 (19.4%) of the cases in the topical steroid cream group, respectively. The success rate for the topical steroid cream was 69.4% and 63.9% at 3 and 8.3 months, respectively. The objectivation of the phimosis grade did not predict the outcome (P > .05). No side effects were associated with the topical steroid treatment. CONCLUSION: The pretreatment classification of phimosis did not allow the prediction of success with the topical steroid treatment. We believe that topical steroid therapy with foreskin retraction and daily cleansing is a valid therapy modality that should be offered before any surgical intervention, regardless of the degree of phimosis.

Deletion of one SERCA2 allele confers protection against bladder wall hypertrophy in a murine model of partial bladder outlet obstruction.

The sarco(endo)plasmic reticulum Ca(2+)-ATPase2 (SERCA2) is downregulated in cardiac hypertrophy with decompensation. We sought to determine whether mice heterozygous for the SERCA2 allele would develop greater bladder hypertrophy and decompensation than their wild-type littermates following partial bladder outlet obstruction (pBOO). We found that following 4 wk of surgically created pBOO, SERCA2 heterozygous murine bladders showed significantly less hypertrophy, improved in vitro cystometry performance, diminished expression of the slow myosin isoform A analyzed by RT-PCR, a significant drop in nuclear translocation of nuclear factor of activated T cells by EMSA, and decreased cell proliferation within the smooth muscle layer following 5-bromo-2'-deoxyuridine labeling compared with their wild-type littermates. Thus, in contrast to cardiac muscle, deletion of a SERCA2 allele confers protection against bladder hypertrophy in a murine model of pBOO. Compensatory mechanisms in heterozygous mice seem to be related to the calcineurin pathway. Further studies are underway to better define the molecular basis of this observation, which has potential clinical applications.

Sexual function in adult patients with spina bifida and its impact on quality of life.

PURPOSE: We evaluated sexual function in adult patients with spina bifida and its impact on quality of life. MATERIALS AND METHODS: Between March 2005 and May 2006, 76 adult patients with spina bifida, including 34 women and 42 men, with a mean age of 24.4 years (range 18 to 37) were seen for followup at our institution. Following institutional review board approval data were collected from medical records and validated questionnaires were completed, including the Watts Sexual Function Questionnaire to assess sexual function and SF-36 to assess quality of life. RESULTS: Of the 76 patients 18 (24%), including 9 women and 9 men, achieved sexual intercourse at least once in the last 2 months. There was no difference regarding gender distribution and mean age +/- SD in sexually active vs not sexually active patients (25.8 +/- 4.2 vs 24 +/- 4.5 years, p = 0.13). All levels (thoracic to sacral) of myelomeningocele were seen in the 2 groups with significant higher lesions of neurological impairment in not sexually active than in sexually active patients. No difference was seen in relation to ambulatory status and urinary incontinence. Overall total Watts Sexual Function Questionnaire scores in sexually active patients were similar in men and women. When comparing the 4 domain scores on desire, arousal, orgasm and satisfaction, women scored similar to men. Only 2 men tried to achieve paternity but failed and 1 woman gave birth. When evaluating SF-36 for quality of life, there was no significant difference in physical health (42.4 +/- 11.9 vs 38.7 +/- 7.2, p = 0.11) and mental health (54.1 +/- 11.3 vs 58.6 +/- 10.7, p = 0.12) subscores in sexually active vs not sexually active patients. CONCLUSIONS: In our cohort 24% of adult patients with spina bifida were sexually active. Sexual activity was not related to gender, level of urinary incontinence or extent of physical disability but it was more likely in patients with more caudal levels of neurological impairment. Sexual function seems not to affect health related quality of life in these patients.

Bilateral pelvi-ureteric junction obstruction in a neonate presenting with sudden anuria.

We report an extremely rare case of a female neonate with antenatally diagnosed bilateral moderate hydronephrosis and appropriate postnatal follow up who presented at 6 weeks of life with sudden onset of anuria secondary to bilateral pelvi-ureteric junction obstructions. While most antenatally diagnosed hydronephrosis remains asymptomatic, this case serves as a reminder that neonatal patients can become acutely symptomatic.