8-Bromo-3',5'-adenosine monophosphate stimulates the endocrine activity of human cytotrophoblasts in culture.

Cytotrophoblasts, purified from human term placentae, were cultured in the absence or presence of 8-bromo-cAMP or 8-bromo-cGMP. 8-Bromo-cAMP provoked a dose-dependent increase in the secretion of hCG and progesterone within 24 h. After 48 h, hCG secretion increased by more than 200-fold, and progesterone secretion increased nearly 5-fold. 8-Bromo-cGMP had no effect on hCG secretion. In culture in serum-supplemented medium, the mononuclear cytotrophoblasts aggregated and fused to form syncytia. This morphological transformation was not affected by 8-bromo-cAMP. Immunocytochemical studies of the alpha- and beta-subunits of hCG in control and 8-bromo-cAMP-stimulated cultures demonstrated that the cyclic nucleotide analog promoted the synthesis of both subunits in all cellular forms, including single mononuclear cells, cell aggregates, and syncytia. In serum-free medium, the cytotrophoblasts did not aggregate or form syncytia, yet they responded to 8-bromo-cAMP with an increase in hCG secretion. We conclude that the endocrine function of cytotrophoblasts can be stimulated by a cAMP-dependent mechanism which can be initiated independently of the formation of a syncytium.

Detection of the ovulatory luteinizing hormone (LH) surge with a semiquantitative urinary LH assay.

We examined blood concentrations of ovarian steroid hormones (E2, P) and serum LH concentrations to establish that the Ovu-STICK assay reliably reflects endocrine events in the periovulatory period. In all 16 cycles studied, which included 2 cycles in which the women received CC, the ovulatory surge of LH was detected with the use of the Ovu-STICK assay. In contrast, BBT monitoring correctly predicted the ovulatory event in only half the cycles.

Infertility treatment in women over 40 years of age.

Women experience a sharp decline in fecundity after the age of 40. Most research points to decreasing oocyte competency and ovarian function as the main causes of this downward trend. Aggressive treatment with in vitro fertilization, laboratory techniques that improve the implantation ability of embryos, and ovum donation can help many of these women achieve healthy pregnancies. Such medical advances are accompanied by new ethical challenges.

Arrested puberty associated with a Sertoli-Leydig cell tumor.

Androgen-producing ovarian tumors are rarely recognized as a cause of delayed or arrested puberty, despite their frequent association with secondary amenorrhea in the older patient. A case is discussed of a Sertoli-Leydig cell tumor in an 18-year-old girl resulting in arrest of breast development and primary amenorrhea.

HLA antigen expression and induction by gamma-interferon in cultured human trophoblasts.

The expression and regulation of HLA antigens were examined in isolated human trophoblasts. Immunocytochemical studies that used monoclonal antibodies against class I (HLA-A, B, and C) antigens revealed that both cytotrophoblasts and syncytiotrophoblasts can express HLA heavy chains and beta 2-microglobulin. Expression of these antigens increased with time in culture. The addition of recombinant gamma-interferon (1000 U/ml) to the cultures significantly increased cellular staining for these antigens over controls throughout the 72-hour time course studied. In addition, we noted that gene expression for HLA class I heavy chain was also markedly augmented by the addition of gamma-interferon. Expression of class II (HLA-DR) antigens was not detected in any of the experiments. These results suggest that during in vitro differentiation, cytotrophoblasts can express class I HLA antigens and the genes controlling their production can be regulated. Alterations in the expression or suppression of these antigens during pregnancy could be responsible for some pregnancy complications.

A mucin-like glycoprotein identified by MAG (mouse ascites Golgi) antibodies. Menstrual cycle-dependent localization in human endometrium.

Human endometrial glands synthesize and secrete a high molecular weight mucin-like glycoprotein in a menstrual cycle-dependent fashion. A novel moiety within this Golgi-associated glycoprotein is strongly reactive with IgG antibodies in numerous murine ascites, and has been termed MAG (mouse ascites Golgi). Immunohistochemical staining of 201 endometrial biopsies revealed the following patterns: MAG first appeared in the Golgi on cycle day 5, peaked on day 15, was present on the surface of the luminal epithelium between days 17 and 19, and was no longer detectable after day 19. MAG was also present in cervical, prostate, seminal vesicle, and lacrimal glands, pancreatic acinar cells, gall bladder and bile duct epithelium, and certain cells of the salivary and sweat glands. Interestingly, only tissues from blood group A individuals exhibited this staining. As a common link among all these cell types is the expression of mucins, we speculated that the MAG epitope could be a mucin-associated blood group A-related epitope. This hypothesis was tested by absorption experiments with a variety of glycoconjugates and erythrocytes and by immunoblots of MAG-rich material. The absorption studies demonstrated that only type III porcine mucin (< 1% sialic acid) and blood type A or AB erythrocytes were able to absorb the anti-MAG antibody. Inasmuch as N-acetyl-galactosamine alone, the terminal blood group A carbohydrate, did not block MAG antibody binding, the MAG epitope appears to involve N-acetylgalactosamine plus other determinants. Immunoblots of endometrial extracts and saliva from blood type A individuals revealed MAG-reactive material with a molecular weight > 200 kd under reducing conditions. Because the MAG epitope appears on the endometrial surface during the purported implantation window, we speculate that mucin-like epitopes could play a role in the earliest apposition phases of conceptus-endometrial interaction.