Antibody to human follicle-stimulating hormone: cross-reactivity with three other hormones.

Antibodies directed against human follicle-stimulating hormone (FSH) were demonstrated in rabbit serum by neutralization of biological activity. Antibodies that bound FSH-(131)I were produced in rabbits and guinea pigs by repeated injections of FSH. By (131)I immunochemical methods, we found that at least 90% of the FSH-(131)I-binding antibody failed to distinguish the four human glycoprotein hormones: FSH, luteinizing hormone, chorionic gonadotropin, and thyrotropin, purified as well as endogenous hormone in plasma. Neither growth hormone, adrenocorticotropin, nor a variety of glycoproteins or animal plasmas were able to react with these antibodies.

An acrylamide gel recombination assay for glycoprotein hormone subunits.

A specific and sensitive assay for recombination of the subunits of hCG and other glycoprotein hormones (LH, FSH, and TSH) is presented. The assay depends on the ability of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) to separate clearly the subunits from native hormone. Iodinated subunit is incubated with its identical unlabeled subunit and complementary subunit. Quantitative displacement of the radioactivity to the native hormone position as followed by gel slicing is used as a direct index of recombination. The assay is reproducible, sensitive to less than one picomole of hormone formed, and offers a simple means of evaluating recombination of glycoprotein chains as distinct from biological activity.

A life-table analysis of pregnancy yield in fixed low-dose menotropin therapy for patients in whom clomiphene citrate failed to induce ovulation.

Forty-nine patients in whom clomiphene citrate failed to induce ovulation were treated for 177 cycles with a fixed low dosage of menotropin. Among these 49 patients, there were 24 pregnancies. Among these pregnancies were two that were multiple and three spontaneous abortions. In only one treatment cycle was there a hyperstimulation syndrome. These patients were divided into three clinical groups: the secondary amenorrheic patient, the oligo-amenorrheic patient, and the patient with poor corpus luteum function. There was no statistically significant difference in the pregnancy rate per month among all groups during the first three treatment cycles (average value, 0.07). However, there was a statistically significant improvement in the pregnancy rate per month in the group with secondary amenorrhea and the group with poor corpus luteum in the last three treatment cycles, as compared with the first three treatment cycles (P = 0.05; average value, 0.75). The oligo-amenorrheic patients, on the other hand, during the last 3 months of treatment, had no statistically significant increase in the pregnancy rate per month. These data suggest that menotropin therapy may have a priming effect. These data do not fit the currently accepted model of a constant pregnancy rate per month for all patients. The data suggest that caution should be exercised before combining patient groups when evaluating the results of menotropin therapy.

Serial beta-subunit human chorionic gonadotropin doubling time as a prognosticator of pregnancy outcome in an infertile population.

Prospective evaluation through the use of radioimmunoassay of the beta-subunit of human chorionic gonadotropin (beta-hCG) in blood samples obtained during the first 30 days of gestation was performed on an infertile population at high risk for pregnancy loss. Four hundred and fourteen samples in 281 pregnancies were analyzed. On the basis of single, random beta-hCG samples in asymptomatic patients, 77% of successful pregnancies and 59% of abortions were correctly identified. On the basis of beta-hCG doubling time (mean 2.2 days +/- 1.0 [2 SD]) computed from serial sampling, again in asymptomatic patients, 88% of successful pregnancies and 76% of abortions were correctly identified. beta-hCG doubling time appears to provide a reliable method of evaluating early pregnancy prognosis with significantly greater ability to identify problem pregnancies within the first 30 days of gestation than does single random hCG values.

Sex selection by sperm separation and insemination.

The Ericsson albumin filtration technique was used to collect a fraction rich in Y sperm for selective insemination in couples desiring a male infant. Of 35 conceptions in which sex was known at delivery or spontaneous abortion, there were 28 males (80%). Twelve pregnancies were achieved after separation of sperm in a Sephadex gel filtration system designed to allow for collection of a fraction enriched in X sperm. Seven pregnancies have resulted in females, two in males, and one in twins of each sex. One patient aborted, and one is still pregnant. While selection for either sex can be done electively, on the basis of sociologic preference, female selection has, as an additional indication, avoidance of male offspring to carriers of sex-linked diseases.

A comparison of serial quantitative serum and urine tests in early pregnancy.

Serial urinary beta human chorionic gonadotropin (beta-hCG) immunoassay was performed on 60 patients during early pregnancy. Results were expressed as tube dilutions positive and were compared with quantitative serum beta human chorionic gonadotropin radioimmunoassay (beta-hCG-RIA) values. The parallel rise of human chorionic gonadotropin (hCG) measured by serum beta-hCG-RIA and a macroflocculation beta-specific urinary immunoassay in early pregnancy was confirmed. The ability of each to predict abortion prior to the onset of patient symptoms or clinical signs of disturbed gestation was quantitated. Results were expressed for each method according to standard regression lines or doubling time for individuals. Utilizing either statistical method, urinary testing was as accurate as serum testing for the prediction of normal pregnancy (about 90% for each). Serum testing was 78% correct in predicting abortion; urinary testing was 63% accurate.

Gonadotropin-releasing hormone radioimmunoassay and its measurement in normal human plasma, secondary amenorrhea, and postmenopausal syndrome.

A sensitive and specific double antibody radioimmunoassay for gonadotropin-releasing hormone (GnRH) has been developed for measurement in ethanol extracts of human plasma. Iodinated hormone was prepared with the use of the chloramine-T method, and antibodies were developed in rabbits over a six-month period with a GnRH synthetic copolymer immunogen. A Scatchard plot revealed at least three species of antibody. The assay can measure conservatively at the 5 pg. per milliliter level and shows no cross-reactivity with other available hypothalamic and pituitary hormones. The releasing hormone was quantitatively recovered from human plasma with immunologic identity to native hormone. Unextracted plasma could not be used because of nonspecific displacement. The measurement of GnRH in individuals receiving 100 mug of intravenous bolus infusions of the synthetic decapeptide show extremely elevated values with two half-lives: one of two to four minutes and another of 35 to 40 minutes. In our experiments, we have found measurable GnRH in patients with secondary amenorrhea and at the midcycle in normal women. In the normal cycling woman during the follicular and luteal phases, GnRH was undetectable. In postmenopausal women with extreme hypoestrogenism and markedly elevated luteinizing hormone values, GnRH was also undetectable. No bursts of GnRH could be detected in normal men when sampled every ten minutes over a two-hour period and every two hours throughout the day.

Increased incidence of aberrant morphological phenotypes in human embryogenesis--an association with endometriosis.

PURPOSE: After noticing specific aberrant forms of development in preimplantation embryos from women with endometriosis, we embarked upon this study in order to examine the frequency of these events as compared with controls. METHODS: A total of 235 embryos representing 30 women undergoing 56 cycles of in vitro fertilization were retrospectively analyzed on videotape and placed into categories based upon the incidence of specific morphological criteria found during observations made on the first and second days following oocyte retrieval. RESULTS: Several of the aberrant nuclear and cytoplasmic events were found to be statistically increased in the group of embryos from women with endometriosis. CONCLUSIONS: The incidence of defined aberrant phenotypes in embryos from women with laparoscopically documented endometriosis may allow us to identify specific embryotoxic events with reference to endometriosis and other diagnoses.

Landmarks during the first forty-two days of gestation demonstrated by the beta-subunit of human chorionic gonadotropin and ultrasound.

Prospective pregnancy evaluation through the combined use of a radioimmunoassay (RIA) for the beta-subunit of human chorionic gonadotropin (beta-hCG) and ultrasound during the first 42 days of gestation after ovulation was performed on a population asymptomatic for first-trimester spontaneous abortion. One hundred forty-six ultrasonic observations in 98 pregnancies were made with simultaneous beta-hCG RIA performed in 80 patients. The following landmarks of normal gestational growth were identified: (1) Before 26 days, beta-HCG RIA permits definitive diagnosis of growing trophoblastic tissue, and serial samples allow doubling time computation for prognosis while ultrasound shows a nonspecific increasing decidual response within the uterus; (2) between 26 and 36 days after ovulation, serial beta-hCG samples continue to give doubling time results while ultrasonic demonstration of a gestational sac is normally seen by 28 days after ovulation; (3) the lack of fetal heart motion by 42 days after ovulation or within a gestational sac with a mean diameter of greater than 30 mm was prognostic of abortion; (4) the absence of a gestational sac by 28 days after ovulation or with a beta-hCG RIA greater than 1,000 ng/ml is suggestive of an ectopic pregnancy until proved otherwise.

Prostaglandin F2a, an ovulatory intermediate in the in vitro perfused rabbit ovary model.

PGF2a has been proposed as a mediator of mammalian ovulation. To elucidate further the role of PGF2a in the process of ovulation, PGF and PGF2a metabolite were measured by radioimmunoassay in the perfusate of an in vitro perfused rabbit ovary preparation. Perfusion medium samples were collected over a 10 to 12 hour period from ovaries perfused with tissue culture M199 (total volume 150 ml, sample volume 3 ml) to which varying amounts of hCG had been added. [The PGF2a antisera a 40% cross reaction with PGF1a, hence total PGF was measured with this antisera.] Both PGF and PGF2a metabolite showed a linear increase with time and numbers of ovulations. PGF media accumulation was 575 pg/ovary/ovulation/hr and PGF2a metabolite accumulation was 367 pg/ovary/ovulation/hr. Medium prostaglandin content could be correlated with numbers of ovulations, ovulatory efficiency (number of ovulations/total follicles) but not total follicles. These data best fit a model of independent ovulatory units producing PGF2a without recruitment or interaction between them. We infer that PGF and PGF2a metabolites in this system can be used as a direct index of the ovulation process.

Inhibitory effect of prolactin on ovulation in the in vitro perfused rabbit ovary.

Prolactin is best known for its effects on the breast, promoting mammary growth and lactation. In some species, including rat, mouse, hamster, sheep and rabbit, prolactin is necessary for the maintenance of the corpus luteum. Further, a relationship has been recognised between hyperpromus and or adenohypophysis. McNatty et al. have postulated a direct ovarian effect of prolactin, reporting that low prolactin levels were essential for progesterone production by preovulatory human granulosa cells cultured in vitro. However, high levels of prolactin decreased progesterone production. The present investigation sought to determine the effect of prolactin on ovulation using the in vitro perfused rabbit ovary preparation. We report that its effects are inhibitory.

Effects of feto-placental markers with plasma exchange in pregnancy.

To evaluate changes in feto-placental markers with plasma exchange in pregnancy, two patients at varying stages of pregnancy referred to a tertiary care hospital and requiring plasma exchange for intercurrent problems were evaluated. Alpha-fetoprotein, human chorionic gonadotropin, and free estriol were sequentially measured in the patients' plasma and in the fluid removed, thus permitting calculations of permeability rates and clearances. Despite markedly different molecular weights, all three feto-placental markers had similar permeabilities and clearances. While in both patients maternal levels of alpha-fetoprotein and human chorionic gonadotropin decreased rapidly with plasma separation and rebounded rapidly to baseline, free estriol responded differently and did not appear to decrease with therapy. Maternal levels of feto-placental markers only transiently changed with plasma exchange during pregnancy and rapidly returned to baseline with no apparent consequences to the pregnancy.