Demonstration of a chorionic gonadotropin-like substance in rabbit morulae.

Rabbit morulae were treated with specific indirect immunofluorescence and immunoperoxidase histochemical techniques. The first antibody in both systems was rabbit anti-human chorionic gonadotropin (hCG) beta-subunit. Negative controls revealed a complete absence of reaction. All morulae incubated with the double-antibody system showed a positive reaction. Rabbit morulae thus present a substance with antigenic determinants similar to the beta-subunit of hCG. The physiologic role of this substance is unknown.     

Presence of a human chorionic gonadotropin--like substance in human sperm

An hCG-like material has been extracted from human sperm. These experiments were designed to characterize this material. Sperms of 10 volunteers were separated from seminal fluid, washed in PBS three times, and resuspended in 0.5 ml of the same buffer. Samples were pooled; cells were disrupted by sonication and extracted in alkaline buffer by constant agitation at 4 degrees C. The extract was ultracentrifuged at 4 degrees C. Supernate was lyophilized and reconstituted in 2 cc of distilled water. This material presented a dose-response curve parallel to those of IS2-hCG and CR119 in beta hCG RIA. When chromatographed in a Sephadex G-150 column the extract eluted within the hCG range and immunoreacted in the specific beta hCG RIA. When absorbed onto a concanavalin A--Sepharose column, all recovered immunoreactive material eluted after exposure to alpha-D-methylglucoside, indicating that it is a glycoprotein. The extract stimulated progesterone and testosterone secretion in porcine granulosa cells and decapsulated rat testis, respectively, indicating its biologic potency.     

Evidence for a human chorionic gonadotropin-like material in the rabbit blastocyst

Extracts from (1) 300 day 6 rabbit blastocysts, (2) 300 day 2 unfertilized ova, and (3) uteri from the nonpregnant does were analyzed in the following assays in order to determine the presence of a chorionic gonadotropin: (1) in vitro bioassay for testosterone production by decapsulated rat testes, (2) in vitro bioassay for progesterone production by porcine granulosa cells, (3) in vitro determination of adenylyl cyclase-stimulating activity in rabbit Graafian follicles, (4) gel filtration in a Sephadex G-150 column and assay of the elutions in a radioimmunoassay (RIA) specific for the beta-subunit of human chorionic gonadotropin (hCG), (5) parallelism with hCG standard preparations in beta-hCG RIA, and (6) concanavalin A-column chromatography. The rabbit blastocyst extracts showed an hCG-like material in all of the systems tested. None of the other two extracts presented hCG-like activity in any of the assay. The immunoreactive material in the beta-hCG RIA of the blastocyst extracts after gel filtration presented a profile different from that of pure hCG; this may represent heterogeneity due to a species difference or a slightly different molecular weight. These results confirmed previous findings of several investigators and those from our laboratory in that the preimplanted rabbit embryo contains a gonadotropin with characteristics similar to hCG.     

Immunocytochemistry of human chorionic gonadotropin in human chorionic villi

The immunocytochemical localization of human chorionic gonadotropin was investigated in chorionic villi from the seventh to twelfth week of gestation. By the light microscopic peroxidase-antiperoxidase technique, positive reactions of human chorionic gonadotropin were found exclusively in the syncytiotrophoblast. Immunoelectron microscopy by means of the protein A-gold technique reveals localization of the immunoreactive gold particles in two kinds of membrane-bound granular inclusions in this cell; one type is granules of 200 to 300 nm in diameter with moderate electron density and the other is large electron-dense bodies of 500 to 1000 nm. The former seems to be Golgi-derived secretory granules that play a role in the release of human chorionic gonadotropin from the syncytiotrophoblast. Although the origin of the latter is still uncertain, a certain amount of this hormone might be stored or treated by lysosomal digestion in the large bodies during these stages.     

Diagnosis of a supernumerary ovary with human chorionic gonadotropin.

The preoperative diagnosis of ectopic ovarian tissue has been reported only once in the literature. In the present case, differential ovarian and adrenal testing was used to diagnose and aberrant ovarian source of persistently elevated plasma estrogen in a patient who had undergone bilateral oophorectomy and adrenalectomy for metastatic breast carcinoma. The ovarian source was diagnosed by stimulation with human chorionic gonadotropin (hCG), and treatment with human menopausal gonadotropin (hMG) prior to surgery was used to facilitate its location at laparotomy.     

Intracytoplasmic sperm injection pregnancy after low-dose human chorionic gonadotropin alone to support ovarian folliculogenesis

OBJECTIVE: To prove that several days of low-dose hCG alone can be used to stimulate folliculogenesis, complete FSH-initiated follicle/oocyte maturation, and achieve pregnancy in assisted reproduction technology. DESIGN: Case report. SETTING: Reproductive endocrinology center at an academic institution. PATIENT(S): A 35-year-old female patient and her partner with male-related infertility. INTERVENTION(S): After an 8-day priming with hMG (225 IU/d), we administered low-dose hCG (200 IU/d) alone for 5 days in one GnRH-agonist suppressed patient until proper follicle development was obtained and intracytoplasmic sperm injection was performed. MAIN OUTCOME MEASURE(S): Daily serum levels of LH, FSH, hCG, E(2), P, and T; measurements of follicle number and size; oocytes retrieved and fertilized; pregnancy. RESULT(S): Although FSH levels rapidly declined after hMG discontinuation, E(2) and large follicles increased during hCG-only administration. Several good quality oocytes were retrieved and fertilized by intracytoplasmic sperm injection; three embryos were transferred and a twin pregnancy ensued. CONCLUSION(S): Replacement of FSH with low-dose hCG for several days in the late ovulation induction stages of assisted reproduction technology resulted in: [1] continued growth of large ovarian follicles and E(2); [2] an optimal preovulatory follicle pattern consisting of many large and few medium and small follicles; and [3] reproductively competent oocytes and pregnancy.     

Significance of sperm antibodies in human fertility.

Indirect immunofluorescent, macroagglutination and immobilizing sperm antibody estimations were performed as part of a prospective study on a representative group of 63 infertile couples. Serum from both, cervical mucus and seminal plasma were tested and the clinical use of the tests evaluated by considering the pregnancies that occurred and the causes of infertility that were discovered in the subsequent year. The indirect immunofluorescent test had the largest number of positives although more were found in seminal plasma than elsewhere. In these, both male and cervical factors were implicated and in this media there were less positives pregnant with the acrosome and midpiece patterns absent. The macroagglutination test had no positives pregnant and again more significance was attached to positives in the male. Similarly, immobilization results showed most positives in seminal plasma where the male and cervical factors featured prominently. It would, therefore, appear that if any clinical use is to be derived from the three tests in the search for antisperm antibodies as a cause of infertility, testing the male and especially seminal plasma appears to correlate best. If treatment is to be recommended, therefore, it should be concentrated on the man.     

Serum human chorionic somatomammotropin in unaborted hydatidiform mole.

Serum human chorionic somatomammotropin (hCS) was measured in 35 patients with intact unaborted hydatidiform mole by a rapid radioimmunoassay using 70% dioxane in water to separate the bound from unbound fraction. Serum hCG was measured by a hemagglutination inhibition method. Serum hCS ranged from 250 to 5900 ng/ml, while serum hCG ranged from 60 IU/ml on unaborted molar pregnancies. Serum hCS in hydatidiform mole increases from a mean +/- SE of 650 +/- 88.5 ng/ml at 7 to 9 weeks' gestation to 1986.7 +/- 859.3 ng/ml at 22 to 25 week's gestation. There was a significant correlation between uterine size and serum hCS in molar pregnancies (correlation coefficient r = +0.5183; P = 0.0025). There was no significant correlation between serum hCS and serum hCG. Serum hCS in a patient with molar pregnancy who subsequently developed choriocarcinoma was not significantly different from that in patients who did not. The findings indicate: 1) that peripheral hCS increases with increased gestational age in molar pregnancies, 2) that the amount of peripheral serum hCS is related to the mass of molar tissue present and not dependent on serum hCG level, and 3) that the serum hCS level in unaborted hydatidiform mole was a poor index for predicting malignant sequelae.     

Daily rhythm of serum human chorionic gonadotropin and human chorionic somatomammotropin in normal pregnancy

We have quantified the human chorionic gonadotropin (HCG) and human chorionic somatomammotropin (HCS) serum levels in normal pregnant women at 1-h intervals during 24 h, through radioimmunoassay procedures. The HCG levels were higher during the first trimester, although no circadian variations were detected. Some variations exceeded by ten fold; these variations decreased with advanced periods of gestation. No diurnal fluctuations in serum HCS were apparent; in a case nearly at term, there was an increase from 05:00 h to 09:00 h simulating a circadian pattern, although it was not statistically significant. These data indicate that the placental protein secretion is autonomous and that HCG levels of fluctuation during early stages of pregnancy might be regulated by factors other than the trophoblast.     

The optimal time for intracytoplasmic sperm injection in the human is from 37 to 41 hours after administration of human chorionic gonadotropin

Fertilization rates increased continuously with the time elapsed after administration of hCG, reaching a peak of 84% when intracytoplasmic sperm injection (ICSI) was performed >41 hours after hCG administration. However, the highest implantation rate, 24%, was achieved when ICSI was performed 37-41 hours after hCG administration.     

Serum hyperglycosylated human chorionic gonadotropin to predict the gestational outcome in in vitro fertilization/intracytoplasmic sperm injection pregnancies

Hyperglycosylated human chorionic gonadotropin (H-hCG) is secreted by the placenta in early pregnancy. Decreased H-hCG levels have been associated with abortion in spontaneous pregnancy. We retrospectively measured H-hCG and dimeric hCG in the sera of 87 in vitro fertilization patients obtained in the 3 weeks following embryo transfer and set the results in relation to pregnancy outcome. H-hCG and dimeric hCG were correlated (r(2) = 0.89), and were significantly decreased in biochemical pregnancy (2 microg/l and 18 IU/l, respectively) compared to early pregnancy loss (22 microg/l and 331 IU/l) and ongoing pregnancy (32 microg/l and 353 IU/l). Only H-hCG tended to discriminate between these last two groups.     

Application of a radioreceptorassay of human chorionic gonadotropin in the diagnosis of early abortion.

A radioreceptorassay for human chorionic gonadotropin (hCG) was used to detect pregnancy as early as 6 to 8 days following conception and to predict spontaneous abortion as early as 8 to 10 days after conception in subjects whose hCG levels were lower than those occurring during normal pregnancy of the same duration. The evidence for the specific measurement of hCG was further provided by the correlation of plasma hCG levels measured by radioimmunoassay using antisera specific for hCG-beta and radioreceptorassay of hCG. By virtue of its sensitivity and specificity, the radioreceptorassay of hCG allowed early and accurate prediction of spontaneous abortion in 15 patients.     

Testicular origin of immunobead-reacting antigens in human sperm.

OBJECTIVE: To investigate, by means of immunobinding technique, the antigenic surface expression of human vasa efferentia sperm as compared with that of ejaculated sperm. DESIGN: Briefly, vasa efferentia sperm, retrieved microsurgically, and donors' ejaculated sperm (controls) were used. PATIENTS, PARTICIPANTS: Men with congenital absence of the vas deferens, undergoing sperm aspiration as part of their infertility treatment, and controls who were donors of our sperm bank were first assayed by direct immunobead test and, if found negative, were exposed to sera containing known high titers of antisperm antibodies and then retested by indirect immunobead test. RESULTS: The data showed no difference in binding titers and class of immunoglobulins between vasa efferentia and ejaculated sperm. CONCLUSIONS: These findings suggest that, in humans, sperm acquire those surface antigens commonly detected by immunobinding test within the testes before their transit through the epididymis.     

Experience of human chorionic gonadotropin assays in gestational trophoblastic tumors.

From May 1979 through December 1988, 146 patients with gestational trophoblastic tumors (71 hydatidiform mole, 3 partial mole, 15 choriocarcinoma and 57 persistent trophoblastic tumors) were studied. A total of 1178 daily urine samples were collected before and/or after treatment, and in the course of follow-up. H93 RIA (an HCG specific assay), H80 RIA (an assay detecting hCG and hLH) and a hCG alpha assay measured levels in the urine specimens. Three hCG declining patterns (pattern D, P and R) based on the H93 RIA assay were noted. Patients showing pattern D had the most favorable outcome (no mortality at all). However, pattern P and R had a 10% and 14.3% mortality rate, respectively. The ratios of H80/H93, hCG alpha/H93, hCG alpha/H80 in the urine specimens were similar in both pattern D and R (excluding samples from a patient who expired later). However, the ratios of H80/H93, hCG alpha/H93, hCG alpha/H80 of samples from the patient (CK) who expired later were significantly different from those of the pattern D and R. This was suggestive of a marked unbalanced secretion of hCG and its subunit in the urine specimens of patient CK. The molecular forms in pattern D were similar to the standard hCG. However, the molecular form in pattern R of 3 fatal choriocarcinomas showed a great variation, from smaller to larger than the standard hCG. The isoelectric points of hCG in pattern D and R were all acidic. In clinical practice, we can measure the ratios of H80/H93, hCG alpha/H93 and hCG alpha/H80, molecular forms, and isoelectric points of hCG.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of human chorionic gonadotropin preparations on complement in vitro.

The effects on complement in vitro by several commercial preparation of human chorionic gonadotropin (HCG) were investigated. These preparations were demonstrably anticomplementary and produced marked conversion of the third component of complement (C'3). However, detailed analyses showed that the anticomplementary activity was because of the presence of immunoglobulins in the HCG preparations. In view of the known modifying effects of C'3-reactive agents on skin allograft survival and of antigen-antibody complexes on lymphocyte transformation, it may be necessary to reconsider previous work on the immunosuppressive effects of HCG preparations.