Antiproliferative effects of low-dose micronized progesterone.

Objective: To study the pharmacodynamic effects of oral micronized P on endometrial maturation.

Design: This was a controlled, open, parallel group, pilot study.

Setting: The experiment was performed in an outpatient academic clinical research unit.

Inhibition of ovulation by progestin analogs (agonists vs antagonists): preliminary evidence for different sites and mechanisms of actions.

Continuous administration of the antiprogesterone RU486 inhibits ovulation in women and in monkeys; in this regard RU486 may act as a progestin agonist rather than as an antagonist. We compared the site(s) and mechanism(s) of RU486-induced ovulation inhibition with those of levonorgestrel (LNG). Six regularly menstruating cynomolgus monkeys each received placebo, RU486 (1 mg/kg/d) or LNG (2 g/kg/d) i.

Use of a specific zona pellucida (ZP) protein 3 antiserum as a clinical marker for human ZP integrity and function.

Objective: To evaluate binding characteristics of a specific zona pellucida (ZP) protein 3 (ZP3) antiserum to human oocytes in order to determine its usefulness as a clinical marker for human ZP integrity and function and its correlation with IVF outcome.

Design: Prospectively designed, blinded, internally controlled study.

Setting: Tertiary care academic center.

Primate reproductive organs reveal a novel pattern of proto-oncogene c-mos and transcription factor Oct-3 mRNA expression.

In mice, expression of the transcription factor Oct-3 and the proto-oncogene c-mos is limited to germ cells, suggesting a specific role for these factors in gamete physiology and early embryonic development. We have studied the expression pattern of Oct-3 and c-mos in various reproductive as well as control tissues in the cynomolgus monkey, using reverse transcriptase polymerase chain reaction (RT-PCR) and Northern analysis. Analogously with the data from the mouse model, strong expression of Oct-3 and c-mos could be detected in monkey ovary and oocytes.

The role of endothelin-1 in regulating human granulosa cell proliferation and steroidogenesis in vitro.

The effects of endothelin-1 (ET-1) on luteinized human granulosa cells (L-HGCs) have not been examined. It is well known that there are differences of actions of several autocrine/paracrine regulators between L-HGCs and GCs of other species, and therefore the present study was designed to examine the effects of ET-1 1) on intracellular Ca2+ concentrations ([Ca2+]i) using the Ca(2+)-responsive fluorescent indicator Fura-2, 2) on cell proliferation by the nonradioactive method using bromodeoxyuridine, and 3) on basal and gonadotropin-stimulated steroidogenesis, and to examine the expression of ET receptor messenger RNA (mRNA) using freshly isolated and cultured L-HGCs obtained from patients undergoing in vitro fertilization. ET-1 increased [Ca2+]i in L-HGCs in a dose-dependent manner between 1 and 1000 nmol/L.

Pharmacokinetics of a single dose of buccal testosterone.

The bioavailability, pharmacokinetics, and metabolism of a novel transbuccal delivery system of testosterone was investigated in five healthy eugonadal men. Total serum testosterone (T), dihydrotestosterone (DHT), and sex hormone-binding globulin (SHBG) concentrations were determined from blood samples obtained at 8:00 a.m.

Intracytoplasmic sperm injection: achievement of high pregnancy rates in couples with severe male factor infertility is dependent primarily upon female and not male factors.

Objective: To determine the efficacy and factors affecting outcome of intracytoplasmic sperm injection (ICSI) in patients with severe male factor infertility.

Design: Prospectively designed clinical trial of patients selected to participate in the study based upon the following inclusion criteria: previous total failed fertilization or unsuitable sperm parameters for conventional IVF.

Setting: Tertiary care academic center.

Prediction of ovarian response to exogenous gonadotropin stimulation: utilization for collection of primate oocytes for fertilization in vitro.

The predictive value of 2 tests for ovarian response to controlled ovarian hyperstimulation in the cynomolgus monkey model was evaluated. The tests utilized were: 1) the cycle Day 3 (Day 1 = onset of menses) FSH value and 2) the acute estradiol (E(2)) response to a GnRH agonists (GnRHa) administered on Day 3. Both tests were performed during the cycle preceding control ovarian hyperstimulation.

An update on the laboratory assessment of male fertility.

Contemporary methods of assisted reproduction have undoubtedly enhanced the opportunities for conception in couples suffering from male infertility. The causes of male infertility are multiple, and current therapies do not usually address an aetiological or pathophysiological origin. Consequently, there is a fundamental need to gain knowledge for optimization of the clinical management of male infertility.

New concepts in human sperm-zona pellucida interaction.

Binding of spermatozoa to the zona pellucida is an initial, crucial recognition event leading to fertilization. In the mouse, the best species characterized so far, the zona pellucida protein 3 (ZP3) has a central role as the specific, primary sperm receptor on the zona and as the inducer of the acrosome reaction. This sequence of events is not clearly understood as it relates to human gametes.

Increase in androgen:estrogen ratio specifically during low-dose follicle-stimulating hormone therapy for polycystic ovary syndrome.

Objectives: To compare changes in serum androgens in women with polycystic ovary syndrome (PCOS) during ovulation induction with low-dose versus conventional urofollitropin.

Design: Prospective case-control study.

Setting: Tertiary-care reproductive medicine center.

Cryopreservation: the problem of evaluation.

A simple evaluatory formula for reporting pregnancy rates involving cryopreserved material has so far remained elusive. It is highly desirable to have a method which would allow not only an evaluation of cryotechnology per se, but also an evaluation of the role of cryotechnology in enhancing the total reproductive potential of a single cycle. In this paper, nine separate formulae are described which can be used for the evaluation of cryopreservation.

Evaluation of the meiotic spindle apparatus in oocytes undergoing intracytoplasmic sperm injection.

Objective: To determine whether intracytoplasmic sperm injection has an adverse effect on the meiotic spindle of oocytes after injection of a sperm into the ooplasm.

Design: Hamster oocytes were injected with human sperm (Test group) and evaluated for meiotic spindle and chromosome morphology using immunofluorescent staining. Results were compared with control uninjected oocytes exposed to the microscope environment (CS group) and untreated oocytes remaining in the incubator (CI group).

High doses of gonadotropins have no detrimental effect on in vitro fertilization outcome in normal responders.

Objective: To examine the effect of high doses of gonadotropins on IVF outcome in normal (intermediate) responders.

Design: Retrospective analysis of patients undergoing IVF therapy from 1990 to 1994 at our institution.

Setting: Academic tertiary center.

Impact of immunoglobulin isotype and sperm surface location of antisperm antibodies on fertilization in vitro in the human.

Objective: To investigate the impact of immunoglobulin (Ig) isotypes and their location on the human sperm surface on fertilization in vitro.

Design: Restrospective review and analysis.

Setting: Reproductive endocrine division of a level 3 academic center.

Value of sperm morphology assessed by strict criteria for prediction of the outcome of artificial (intrauterine) insemination.

The purpose of this study was to investigate the relationship between sperm parameters and the outcome of artificial (intrauterine) insemination (IUI). One hundred and twenty-six patients undergoing 395 consecutive IUI cycles in a 1-year period in our institution were studied. In all cases, controlled ovarian stimulation and hCG-timed IUI were performed, followed by progesterone supplementation of the luteal phase.

Lactational anovulation in non-human primates: restriction of nursing inhibits Prl secretion without precipitating the return of ovulatory menstrual cyclicity in cynomolgus monkeys.

To test the hypotheses that nighttime suckling and elevated nocturnal prolactin concentrations are essential for the continuation of lactational anovulation, the effects of restricting nursing to twelve h per day (either day or night) on maternal prolactin (Prl) levels and resumption of ovulatory menstrual cycles were studied in five groups of cynomolgus monkeys: Group 1: baby weaned: Group 2: baby fully nursed: Group 3: baby nursed night only; Group 4: baby nursed day only; and Group 5: baby housed with mother but not allowed to nurse. Restrictions were initiated at approximately 150 days postpartum and were achieved by placing a non-nursing jacket over the nursing jacket, which was worn 24 h/day. Fifteen out of seventeen monkeys remained anovulatory while housed with their infants, irrespective of the type of nursing restriction.

Strategies to respond to polymerase chain reaction deoxyribonucleic acid amplification failure in a preimplantation genetic diagnosis program.

Objectives: Our purpose was to identify and evaluate practical methods within a preimplantation genetic diagnosis program that will increase the percentage of embryos for which a genetic diagnosis can be obtained, including clinical responses after failure of deoxyribonucleic acid amplification has occurred.

Study Design: Known human lymphoblast cell lines and human embryo blastomeres were evaluated in a single-cell, nested primer polymerase chain reaction system with primer sequences for the specific locus surrounding the four base pair insertion mutation on exon 11 of beta-hexosaminidase A-Tay-Sachs disease, the delta F508 mutation of cystic fibrosis, and the sex-determining region on the Y chromosome. Reamplification polymerase chain reaction with standard polymerase chain reaction and primer extension preamplification was performed in deoxyribonucleic acid preparations after previous polymerase chain reaction amplification attempts had resulted in failure of amplification.

Estrogen and progesterone receptor mRNA are expressed in distinct pattern in male primate reproductive organs.

Purpose: The role(s) of estrogens (E) and progesterone (P) in male reproductive physiology remain unclear. Estrogens are used in the treatment of prostatic cancer. Progestins have been used to control excessive sexual behavior in men, and proposed as a male contraceptive.

Low-dose glucocorticoids after in vitro fertilization and embryo transfer have no significant effect on pregnancy rate.

Objective: To determine the effect on pregnancy rate (PR) of low-dose glucocorticoid treatment in cycles without micromanipulation.

Design: Randomized, prospective, double-blinded, placebo-controlled trial.

Setting: One university-based tertiary infertility center and two private infertility centers.

Cell cycle genes c-mos and cyclin-B1 are expressed in a specific pattern in human oocytes and preimplantation embryos.

Little is known about the molecular mechanisms governing the development of human oocyte and pre-embryo. We characterized the expression pattern of c-mos, cyclin B1 and beta-actin mRNA in oocytes and granulosa cells from human and monkey, and in human early embryos, using both qualitative and semi-quantitative reverse transcriptase polymerase chain reaction. The proto-oncogene c-mos was expressed in an oocyte-specific manner and no mRNA for c-mos could be detected in the granulosa cells.

Factors affecting fertilization: endometrial placental protein 14 reduces the capacity of human spermatozoa to bind to the human zona pellucida.

Objective: To examine whether placental protein 14 (PP14) may affect directly those sperm functions crucial to fertilization and early embryo development.

Design: In these prospective studies, we evaluated semen samples of fertile men incubated under capacitating conditions with and without PP14.

Setting: Academic tertiary institution.