Michael J. Gast

Tumor necrosis factor in peritoneal fluid of women undergoing laparoscopic surgery

The level of tumor necrosis factor (TNF) in peritoneal fluid (PF-TNF) of 74 women undergoing laparoscopy was determined. The difference between the mean concentration of PF-TNF of women with normal pelvic anatomy and women with moderate or severe endometriosis was significant (P less than 0.01). The proportion of PF-TNF-positive women with PID and those with moderate or severe endometriosis was also significantly higher when compared to women with normal pelvic anatomy (P less than 0.05; P less than 0.02). The proportion of PF-TNF positive women among nulligravid and nulliparous women was significantly higher than that of women with two or more pregnancies (P less than 0.01) and two or more deliveries (P less than 0.005). These results indicate that the presence of PF-TNF is associated with primary infertility and endometriosis.

Endometrial thickness is a valid monitoring parameter in cycles of ovulation induction with menotropins alone

OBJECTIVE To evaluate the ability of an ultrasound (US)-measured periovulatory endometrial thickness to predict conception in hMG-stimulated cycles. DESIGN Retrospective. SETTING A university-based tertiary practice. PATIENTS One hundred twelve patients undergoing 292 cycles of ovulation induction with hMG alone. MAIN OUTCOME MEASURES A periovulatory transvaginal US measurement of endometrial thickness was obtained during cycles of ovulation induction with hMG alone. Clinical pregnancy was defined by fetal cardiac activity. Sensitivity and false-positive rates for multiple discriminatory values of endometrial thickness were calculated and a relative operating characteristic (ROC) curve was constructed to evaluate the performance of this test as a predictor of pregnancy. RESULTS Thirty-eight of 292 cycles resulted in pregnancy. Conception and nonconception cycles showed similar demographics, diagnoses, peak E2, maximum number of follicles, midluteal P, and mean endometrial thickness. Ovulatory dysfunction was a more frequent diagnosis in the conception group. Relative operating characteristic analysis for endometrial thickness as a predictor of pregnancy yielded an area under the curve of 0.623 +/- 0.049 (mean +/- SD). CONCLUSION Endometrial thickness is a valid screening test for conception outcome in cycles stimulated with hMG. A periovulatory endometrial thickness > or = 10 mm defined 91% of conception cycles. No pregnancy occurred when the endometrium measured < 7 mm.

Isolation and sequencing of a complementary deoxyribonucleic acid clone encoding human placental 173-estradiol dehydrogenase: Identification of the putative cofactor binding site

17 beta-Estradiol dehydrogenase (EC 1.1.1.62) catalyzes the interconversion of estradiol and estrone in human term placenta. We have raised a specific polyclonal antibody to this abundant placental enzyme to study its role in late pregnancy events and its molecular biologic characteristics. In this work the 17 beta-estradiol dehydrogenase antibody was used to isolate and sequence a complementary deoxyribonucleic acid clone encoding about 98% of the amino acid sequence of the 17 beta-estradiol dehydrogenase molecule. This sequence verifies previous sequence data on the molecules steroid binding site and also localizes a putative nicotinamide adenine dinucleotide binding region similar to that of many other pyridine nucleotide-dependent dehydrogenases. Isolation of the complementary deoxyribonucleic acid for 17 beta-estradiol dehydrogenase expands our knowledge of the structure-function relationships of the enzyme and is a major step in our understanding of its biologic function in pregnancy.

Mullerian adenosarcoma of the cervix with heterologous elements: Diagnostic and therapeutic approach

Mullerian adenosarcoma of the cervix with heterologous elements is an extremely rare tumor first described by Roth and colleagues (L. M. Roth, G. L. Pride, and H. M. Sharma, Cancer 37, 1725-1736) in 1976. Since that time there have been only three subsequent reports of these cervical Mullerian adenosarcomas, which seem to occur most often in the postmenarchal age group. Due to the paucity of cases and the unknown biological potential of the tumor, therapy has ranged from simple excision to radical pelvic surgery and vaginectomy combined with both radiotherapy and chemotherapy. We report another case of Mullerian adenosarcoma of the cervix occurring in a teenage woman and make recommendations about diagnostic and therapeutic measures available to the physician.

Long-term follow-up of couples after hamster egg penetration testing

OBJECTIVE To determine the clinical usefulness of the zona-free hamster egg penetration test as a long-term prognostic indicator for future pregnancy. SETTING Division of Reproductive Endocrinology and Infertility at the Washington University Medical Center. PARTICIPANTS All couples (n = 148) who had a hamster egg penetration assay performed between March 1, 1985 and December 31, 1986 were identified and followed with direct or telephone contact up to 68 months after the initial assay. MAIN OUTCOME MEASURE The monthly fecundity rates using life table analysis and the 5-year incidence of pregnancy were categorized by the percentage of hamster eggs penetrated and by history of previous urologic surgery. RESULTS There were no significant differences in the rate nor incidence of pregnancy in couples with hamster egg penetration scores of 0%, > 0% and < or = 10%, > 10% and < or = 20%, or > 20%. Although men with previous urologic surgery tended to have lower scores, there was no significant difference in the 5-year incidence of pregnancy. CONCLUSION The hamster egg penetration score is not predictive of incidence of pregnancy nor time to conception.

Does intrauterine insemination offer an advantage to cervical cap insemination in a donor insemination program

OBJECTIVE To compare pregnancy outcome after IUI versus cervical cap insemination in a donor insemination program. DESIGN A randomized prospective clinical trial in which patients were alternately inseminated with cryopreserved human semen using either IUI or cervical cap insemination methods. SETTING The donor insemination program at Washington University School of Medicine. PATIENTS Forty-two women with either isolated male factor or male factor plus corrected ovulatory dysfunction using clomiphene citrate underwent 141 cycles of donor insemination. MAIN OUTCOME MEASURES Clinical pregnancy rates (PRs) defined as a viable intrauterine gestation > 12 weeks or delivered were compared between groups using the chi 2 test. RESULTS Clinical PRs were significantly higher in the IUI group (16.4%) compared with the cervical cap insemination group (5.9%). The spontaneous abortion rates were similar between the IUI (1.4%) and cervical cap insemination groups (4.4%). CONCLUSIONS These findings suggest an advantage to IUI over cervical cap insemination in a donor insemination program.

STUDIES ON LUTEAL GENERATION AND PROCESSING OF THE HIGH MOLECULAR WEIGHT RELAXIN PRECURSOR

The biology of reproductive control is of special interest to the basic and clinical scientist because of the far-reaching consequences of the pharmacologic regulation of the processes of fertilization and gestation. The ovary occupies a central position in the delicately balanced system of hormonal signals that insure the successful, repetitive reproductive outcomes necessary for species maintainence. Although steroid production by the ovary has been extensively studied, little is known of the mechanics of the ovarian elaboration of secretory proteins. Because relaxin is the only well-characterized ovarian secretory protein, our interest in ovarian maturation and gene expression led us to use relaxin synthesis as a tool to study these ovarian cellular functions. To do so, we utilized translational assay of ovarian RNAt in a reconstituted ascites tumor cell-free system.l In 1978 we showed that RNA isolated from corpora lutea of pregnant sows directs the synthesis in vitro of a 23,000 molecular weight (MW) protein with immunologic and sequence identity to authentic relaxin.*, This relaxincontaining-protein (or RCP) is the primary translation product of ovarian relaxin biosynthesis. We have since shown that RCP is a single chain molecule containing all of the cysteine-bearing peptides of relaxin and is structurally similar to the preproinsulin m01ecule.~ It is likely that a series of maturational modifications occur to the relaxin precursor. Such modifications may be an important part of the process of synthesis, storage, and release that are responsible for relaxin’s delivery to its target tissues. They probably occur at several subcellular sites. We chose to examine prepeptide cleavage and membrane translocation, the initial processing events responsible for the generation of mature relaxin from RCP. Because relaxin is a representative ovarian protein we were interested in evaluating the relationship between the cellular levels of the messenger RNA for RCP and

Therapeutic donor insemination: a prospective randomized study of scheduling methods.

OBJECTIVE To compare basal body temperature (BBT) graphs and urinary luteinizing hormone (LH) monitoring in scheduling therapeutic donor insemination. DESIGN Participants were prospectively randomized to the BBT or LH groups. SETTING Participants were private patients of the Reproductive Endocrine Division at Washington University School of Medicine. PATIENTS Inclusion criteria were designed to assure an isolated male factor. Seventy-four of 113 patients completed the study; 18 had ongoing treatment at the end of the study. INTERVENTIONS Basal body temperature graphs were physician interpreted and appointments prospectively chosen. Luteinizing hormone patients monitored daily urine samples and scheduled an appointment the day after the detected surge. MAIN OUTCOME MEASURES Fecundity rates, cumulative pregnancy rates, and cost per pregnancy were all prospectively evaluated. RESULTS Life table analysis yielded a 6-month cumulative probability of pregnancy of 36.3% in the LH group and 65.1% in the BBT group (P less than 0.025). The total cost per pregnancy was lower in the BBT group (+6,212 versus +3,997; P less than 0.001). CONCLUSIONS This randomized prospective study demonstrates significant therapeutic and economic advantages when therapeutic donor insemination is prospectively scheduled by BBT graphs.

Regulation of Creatine Kinase Isoenzymes in Human Placenta During Early, Mid-, and Late Gestation:

Objective: Creative kinase (CK) isoenzymes play an important role in cellular energy transduction. Two isoenzymes of creatine kinase, ubiquitous mitochondrial creatine kinase (uMtCK) and cytosolic brain creatine kinase (BCK), are postulated to form the creatine phosphate (CP) shuttle, in which creatine serves to transport high-energy phosphate from the mitochondria to its site of utilization. Coordinate regulation of these genes is essential for efficient energy transduction. We examined human CK isoenzyme regulation in placentas during all three trimesters of gestation to define the mRNA and protein expression patterns of uMtCK and BCK and to test the CP shuttle hypothesis. Methods: Placental samples were collected from a total of 26 patients from the first, second, and third trimesters. Total RNA and protein were prepared from each sample and quantified. Quantitative RNA analysis was performed by gel electrophoresis and dot blot techniques using isoenzyme-specific human cDNA probes for uMtCK and BCK. Protein expression of uMtCK and BCK was examined by Western blot analysis using isoenzyme-specific antibodies to uMtCK and BCK. Results: Analysis of RNA demonstrated the coordinate expression of uMtCK and BCK mRNAs in human placenta, with peak expression ofboth in the term placentas. Western blot analysis demonstrated coordinate expression of uMtCK and BCK proteins in the first and second trimesters, but not in the term placenta. Expression levels of uMtCK and BCK proteins were not consistent with their respective mRNA levels in the term placenta. Conclusion: Expression of uMtCK and BCK in human placenta is highly regulated, and posttranscriptional regulation of uMtCK and BCK expression occurs in the term placenta. The coordinate regulation of uMtCK and BCK in human placenta supports the CP shuttle hypothesis. This analysis demonstrates that human placenta has high energy needs that can change rapidly; thus, a functioning CP shuttle may be important in the maintenance and termination of pregnancy.

Evolution of clinical agents for ovulation induction

Since the first use of human urinary gonadotropins in the 1960s, the number of agents available for ovulation induction has remained fairly static. However, the nature of gonadotropin therapy is expected to change rapidly in the near future, and by the end of the decade these changes may be augmented by nongonadotropin ovulation induction therapies. Newer agents described in the literature include highly purified preparations of follicle-stimulating hormone and recombinant forms of follicle-stimulating hormone, luteinizing hormone, and human chorionic gonadotropin. Protein fragments, cytokines, and growth factors also show great promise as ovulation adjuncts. Other promising approaches being explored are the use of genetically engineered human gonadotropin derivatives, the creation of chimeric proteins, and gene therapy.