M. DiMattina

Effect of the antiprogestin RU486 on progesterone production by cultured human granulosa cells: Inhibition of the ovarian 3B-Hydroxysteroid dehydrogenase

Recent studies suggest that the antiprogestin RU486 may have a direct effect on human ovarian luteal function. To further examine this possibility, we have studied the effect of RU486 on ovarian steroidogenesis using human granulosa cells obtained from women undergoing in vitro fertilization. RU486 at concentrations of 0.1, 2, 5, 10 and 100 nM was incubated with 10(5) granulosa cells over 72 hours. Significant suppression of progesterone production occurred following treatment of cultured cells with 2, 5, 10, and 100 nM RU486 at 24 hours (p less than 0.05) and 48 hours (p less than 0.01). At 72 hours, significant decreases in progesterone production were observed with 10 nM (p less than 0.05) and 100 nM RU486 (p less than 0.01). The greatest effect of RU486 on progesterone production occurred at 24 hours of incubation (slope = -8.03) compared with 48 (slope = -4.71) or 72 (slope = -2.31) hours (p less than 0.01). Maximal suppression of progesterone production occurred using 10 nM RU486 with no further significant suppression observed with 100 nM RU486. Other steroids (R5020, DHA) failed to suppress progesterone production suggesting that the observed inhibitory effect on progesterone was specific to RU486. To better understand how RU486 decreases progesterone production in granulosa cell cultures, we measured human ovarian 3B-hydroxysteroid dehydrogenase (3BHSD) in the presence and absence of RU486 in vitro. A significant dose-dependent decrease in the activity of 3BHSD was observed at concentrations of RU486 that were equal to or greater than substrate concentration. Taken together, these findings suggest that RU486 may directly affect human ovarian progesterone production through a mechanism that involves a reduction in 3BHSD activity.

Utilization and success rates of unstimulated in vitro fertilization in the United States: an analysis of the Society for Assisted Reproductive Technology database

OBJECTIVE To examine the utilization and outcomes of natural cycle (unstimulated) IVF as reported to the Society of Assisted Reproductive Technology (SART) in 2006 and 2007. DESIGN Retrospective analysis. SETTING Dataset analysis from the SART Clinical Outcome Reporting System national database. PATIENT(S) All patients undergoing IVF as reported to SART in 2006 and 2007. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Utilization of unstimulated IVF; description of patient demographics; and comparison of implantation and pregnancy rates between unstimulated and stimulated IVF cycles. RESULT(S) During 2006 and 2007 a total of 795 unstimulated IVF cycles were initiated. Success rates were age dependent, with patients <35 years of age demonstrating clinical pregnancy rates per cycle start, retrieval, and transfer of 19.2%, 26.8%, and 35.9%, respectively. Implantation rates were statistically higher for unstimulated compared with stimulated IVF in patients who were 35 to 42 years old. CONCLUSION(S) Unstimulated IVF represents <1% of the total IVF cycles initiated in the United States. The pregnancy and live birth rates per initiated cycle were 19.2% and 15.2%, respectively, in patients <35 years old. The implantation rates in unstimulated IVF cycles compared favorably to stimulated IVF. Natural cycle IVF may be considered in a wide range of patients as an alternative therapy for the infertile couple.

Ketoconazole inhibits multiple steroidogenic enzymes involved in androgen biosynthesis in the human ovary

Ketoconazole (KZ) has been shown to inhibit testicular and adrenal steroidogenesis and is useful in the medical management of gonadotropin-independent precocious puberty, prostatic cancer, and Cushings syndrome. To determine whether KZ similarly affects ovarian steroidogenesis, the authors examined its effect on the activity of the human ovarian 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta-HSD), 17-hydroxylase (17-OH), and aromatase (AR) in vitro. A dose-dependent decrease in the activities of 3 beta-HSD and 17-OH was observed with increasing amounts of KZ. With 10, 50, and 100-fold excess KZ, the activity of 3 beta-HSD decreased by 59% (P less than 0.001), 73% (P less than 0.005), and 85% (P less than 0.005), respectively. At equimolar concentrations with substrate (50 microM), KZ inhibited 17-OH by 70% (P less than 0.01). No significant effect on ovarian AR activity was observed, except at the highest concentration of KZ tested. The authors conclude that low concentrations of KZ profoundly inhibit the activity of human ovarian 3 beta-HSD and 17-OH in vitro. These observations suggest that KZ might be useful in the medical management of women with hyperandrogenism, but further experimentation and clinical trials will be necessary.

Natural cycle in vitro fertilization implantation rates compared to stimulated in vitro fertilization and role of serum antimullerian hormone levels

Objective: The objective of this study is to compare implantation and singleton live birth rates between natural cycle in vitro fertilization (NCIVF) and stimulated IVF. Stratify the results by age and antimullerian hormone (AMH). Design: Retrospective cohort trial of patients who underwent unstimulated IVF between 2007 and 2011. Stimulated patient data from the 2010 Centers for Disease Control (CDC) report. Setting: Private practice. Patients: Infertility patients Intervention: None. Main Outcome Measures: (1) Implantation rates stratified by age and AMH (2) singleton pregnancy rates. Results: A total of 1288 cycles of NCIVF were compared to 94,976 cycles from CDC. In patients Conclusions: Implantation rates were superior in patients 35-40 undergoing NCIVF compared with stimulated IVF. In NCIVF implantation rate was independent of AMH. The live singleton birth rates per embryo transfer for NCIVF and stimulated IVF are similar.

An improved medium for long-term culture of human embryos overcomes the in vitro developmental block and increases blastocyst formation**Presented in part at the 46th Annual Meeting of The American Fertility Society, Washington, D.C., October 15 to 18, 1990.

OBJECTIVE To determine if a culture medium, designated CZB after the authors who first described it, which is supplemented with 0.11 mM ethylenediaminetetraacetic acid, 1.0 mM glutamine, 31.30 mM lactate, and 0.27 mM pyruvate and is lacking glucose for the initial stages of culture that overcomes the in vitro two-cell block of mouse embryos, can improve the rate of blastocyst formation of human embryos in long-term cultures and increase the pregnancy rate (PR) when used in a clinical in vitro fertilization (IVF) program. DESIGN The study is in two parts. Initially, excess oocytes from IVF and gamete intrafallopian transfer patients were fertilized in vitro and then placed in long-term culture of either CZB plus 10% heat-inactivated human serum (32 zygotes) or Earles balanced salt solution (EBSS) supplemented with 0.45 mM pyruvate plus 10% human serum (28 zygotes) to determine if CZB medium could enhance in vitro development and increase blastocyst formation when compared with EBSS. Subsequently, CZB or EBSS medium was used for short-term cultures of embryos in a clinical IVF program to determine if the use of CZB could increase the clinical IVF PR. SETTING Private practice of one author (M.D.). PATIENTS The excess oocytes were donated by couples not wishing to have cryopreservation. In the clinical trial, 49 couples presenting with tubal or male factor infertility and who had three or more fertilized zygotes were randomly assigned to one of the culture media being used. RESULTS In long-term cultures, embryos were observed at 42, 66, 90, 114, and 138 hours after fertilization and scored for blastomere number, degree of fragmentation, and developmental arrest. When CZB- and EBSS-cultured embryos were evaluated over 138 hours, there was a significant increase in the number of CZB-cultured embryos reaching the blastocyst stage (56% versus 20%; P less than 0.009) and less fragmentation of CZB-cultured embryos (18.8% versus 50%; P less than 0.01). In short-term cultures, pronuclear stage embryos from patients undergoing IVF were cultured in CZB or EBSS for 24 hours, graded, and then used in embryo transfers. Of the 28 patients assigned to EBSS, 6 became pregnant (21.4%), and of the 21 assigned to CZB, 5 attained pregnancy (23.8%). These results were not significantly different. CONCLUSIONS The use of CZB for the long-term culture of human embryos is highly beneficial and increases the rate of blastocyst formation, but its use in an IVF program does not increase the clinical PR.