Ronald D. Wiehle

Differential expression of uterine progesterone receptor forms A and B during the menstrual cycle

Recent studies suggest that the progesterone receptor isoforms (PR-A and PR-B) activate genes differentially and that PR-A may act as a repressor of PR-B function. Hence, the absolute and relative expression of the two isoforms will determine the response to progesterone. We have measured their relative expression in the uterus of cycling women who underwent endometrial biopsy. PR isoforms were identified on blots of SDS-PAGE gels by reaction with the AB-52 antibody after immunoprecipitation from endometrial extract. Both isoforms were highest in the peri-ovulatory phase, but levels of PR-A were always higher than those of PR-B. The ratio of PR-A to PR-B changed during the menstrual cycle. Between days 2 and 8, PR-B is almost undetectable and the A:B ratio is >10:1. From days 9 to 13, the ratio is about 5:1, and it is about 2:1 between days 14 and 16. Thereafter, PR-B dwindles rapidly and is virtually undetectable at the end of the cycle. In various hypoestrogenic environments, PR-B expression was reduced. However, exogenous estrogens in the follicular phase in the form of oral contraceptives, enhanced PR-B expression. These data support the possibility that progesterone acts through cycle-specific PR isoforms.

High doses of oral contraceptives do not alter endometrial α1 and αvβ3 integrins in the late implantation window

Objective To assess the effects of an emergency contraceptive agent on the distribution of integrin heterodimers during that part of the implantation window. Design Prospective, case-controlled study in a university-based Population Program. In the first ovulatory control cycle after the detection of LH surge, patients had endometrial sampling 11 days after the surge. In the next cycle the procedure was repeated 2 days after the administration of a postcoital contraceptive agent on day 9 after LH surge (100 g ethinyl E 2 and 2mg norgestrel). Main Outcome Measures The effects of postcoital contraceptives on the expression of integrin heterodimers ( α 1 and α v β 3 subunits) reported to be unique to secretory phase was determined. Results All six specimens were consistent histologically with days 24 and 25 of the menstrual cycle by light microscopy. Using immunohistochemistry, strong membrane staining of endometrial glandular cells and superficial epithelium for both α 1 subunit and vitronectin ( α v β 3) receptor was observed in treatment and controls. No diminution of intensity or distribution was observed relative to pretreatment controls. Conclusions There is no apparent change in the level of these two integrins in the human endometrium when high-dose oral contraceptives are given in the later stages of the implantation window. This suggests that the high doses of steroids used in emergency contraceptives may exert their effect through more complex mechanisms than endometrial cell surface changes.

Enclomiphene citrate stimulates testosterone production while preventing oligospermia: a randomized phase II clinical trial comparing topical testosterone.

OBJECTIVE To determine the effect of enclomiphene citrate in men with secondary hypogonadism. DESIGN Phase II clinical trial. SETTING Community dwelling men making visits to physician offices. PATIENT(S) Men with secondary hypogonadism. INTERVENTION(S) Oral administration of enclomiphene citrate or 1% topical T gel. MAIN OUTCOME MEASURE(S) Luteinizing hormone, FSH, T, and semen analysis. RESULT(S) Treatment with enclomiphene citrate resulted in increased morning serum T, E2, and LH levels similar to those obtained with a topical T gel in men with secondary hypogonadism. Follicle-stimulating hormone and LH were increased with enclomiphene, and sperm counts were conserved. CONCLUSION(S) Enclomiphene citrate reverses the two hallmarks of secondary hypogonadism, namely, low serum total T and low or inappropriately normal LH while preserving sperm production. CLINICAL TRIAL REGISTRATION NUMBER NCT01270841 (ClinicalTrials.gov Identifier NCT01270841).

Differential effects of isomers of clomiphene citrate on reproductive tissues in male mice

To determine, in a chronic dosing study, the oral toxicity potential of the test substances, enclomiphene citrate (ENC) and zuclomiphene citrate (ZUC), when administered to male mice by oral gavage.

Oral Administration of Enclomiphene Citrate Results in Physiological Total Testosterone Levels in Men with Low or Normal Testosterone: A Pilot Study

Background: Clomiphene Citrate may be used off-label to treat men with secondary hypogonadism. More commonly used and approved are the exogenous testosterone products such as gels, patches, and injectables. One drawback may be the high levels of serum testosterone (TT) achieved with exogenous products. A single isomer of Clomiphene citrate, Enclomiphene citrate, is in development and may be useful and gain regulatory approval. An open question is whether men using Enclomiphene citrate will experience high levels of TT. Aims: In our efforts to evaluate Enclomiphene citrate as an oral therapy for normalizing testosterone, we conducted a Phase I pilot clinical study (ZA-002: An Open Label, Fixed Dose, Single Center, Phase I Study to Evaluate the Changes in Total Testosterone with Oral Administration of Enclomiphene Citrate in Healthy Men with Low and Normal Testosterone) to assess the compounds effects on TT. Methods: Sixteen men, mean age 46, with low or normal testosterone levels were enrolled and 13 administered Enclomiphene Citrate drug for two weeks. The pharmacodynamics effects of Enclomiphene citrate on total testosterone (TT) were investigated. The Phase I study also assessed the safety and tolerability of Enclomiphene citrate in middle aged subjects. Results: An increase in TT was observed in males with low baseline levels following 14 days of Enclomiphene citrate. The mean increase in TT ranged from147 ng/dL to 339 ng/dL and results in an increase into the normal range with few excursions above the upper limits. TT increases were greater in men with normal testosterone at baseline. After 14 days of Enclomiphene citrate, mean testosterone levels over a 24-hour sampling period were relatively similar with respect to Cavg. There was a trough in TT 12 hours after administration. TT levels returned to baseline within 28 days of the last dose of Enclomiphene citrate. No clinically meaningful effects on vital signs, laboratory safety tests or ECG results were noted.

On the mark? Is alkaline phosphatase a surrogate for bone density in men with hypogonadism?

bladder pain. In this model, a high dose of the TRPV4 inhibitor reduced detrusor overactivity, whereas even the high dose of the TRPV1 inhibitor did not; however, a combination of ineffective doses of both inhibitors markedly decreased bladder reflex activity. On the other hand, each of the two drugs caused partial analgesia, but their combination was not more effective than either drug alone. This indicates an interesting functional interaction between TRPV1 and TRPV4 channels, which is specific for the overactivity vs the pain response. Previously, the Cruz group reported that bladder overactivity induced by nerve growth factor depends on the presence of functionally active TRPV1 [4]. Taken together this work shines light on networks of multiple mediators and their receptors that cooperate in the regulation of bladder function but previously have mainly been viewed in isolation. Such work may also have therapeutic consequences. As target-saturating concentrations of ligands at any of these receptors may cause relevant adverse effects, targeting multiple such receptors in low doses may open an avenue for a multi-pronged approach, particularly in patients with bladder dysfunction difficult to control with present treatment options.