The effect of follicular fluid protein(s) on gonadotropin-modulated secretion of progesterone in porcine granulosa cell cultures.

Porcine granulosa cells from medium-sized follicles (2 to 5 mm in diameter) were cultured (100,000 cells per culture) with various concentrations of a 12,500 to 16,000 molecular weight protein fraction isolated from follicular fluid. To these cultures pregnenolone (10(-5) mol/L) and either human chorionic gonadotropin or porcine follicle-stimulating hormone were added. The conversion of pregnenolone to progesterone was used to determine 3-beta-ol-dehydrogenase activity in the granulosa cells. Follicular fluid protein alone caused a biphasic response in the production of progesterone. Follicular fluid protein in the concentration of 167 micrograms/ml caused a tenfold increase in the production of progesterone, whereas the 500 micrograms/ml concentration caused a return to baseline levels. Although porcine follicle-stimulating hormone induced a dose-response increase in the production of progesterone, human chorionic gonadotropin produced no change in the levels of progesterone. Low doses of follicular fluid protein acted synergistically with low doses of porcine follicle-stimulating hormone to increase granulosa cell secretion of progesterone. However, high doses of follicular fluid protein inhibited the low-dose stimulation by porcine follicle-stimulating hormone of the production of progesterone. High doses of porcine follicle-stimulating hormone (10 microgram/culture) overcame both the low-dose enhancement and the high-dose inhibition of follicular fluid protein on granulosa cell secretion of progesterone. Thus follicular fluid protein may be an important paracrine modulator of steroidogenesis stimulated by follicle-stimulating hormone in the porcine granulosa cell.

Effect of suckling on serum prolactin, luteinizing hormone, follicle-stimulating hormone, and estradiol during prolonged lactation.

Eight nursing subjects had serum prolactin (PRL), luteinizing hormone, follicle-stimulating hormone, and estradiol levels studied during the first six months postpartum. Each subject had serum samples obtained just before the initiation of suckling and during the next 120 minutes. Baseline PRL levels were high at ten days postpartum (90.1 ng/mL), then slowly declined but remained elevated at 180 days postpartum (44.3 ng/mL), with the stimulus of suckling being able to double the baseline PRL value throughout the study period. Mean estradiol levels were low at ten days postpartum (7.2 pg/mL), then gradually rose to a mean level of 47.3 pg/mL at 180 days postpartum; however, in four subjects who were amenorrheic during the study period, the mean estradiol levels remained low (4.25 pg/mL), while baseline PRL levels remained high (63.6 ng/mL).

Effect of human menopausal gonadotropin and follicle regulatory protein(s) on 3 beta-hydroxysteroid dehydrogenase in human granulosa cells.

Recently, a protein fraction [follicle regulatory protein (FRP)] which inhibits FSH-induced granulosa cell aromatase activity was isolated from both human and porcine follicular fluid. In this study, the actions of FRP on 3 beta-hydroxysteroid dehydrogenase (3 beta-HSDH) activity were examined using granulosa cells obtained from hyperstimulated patients undergoing oocyte aspiration for in vitro fertilization. Granulosa cells were cultured with 0, 167, or 500 micrograms/ml FRP with or without human menopausal gonadotropin (hMG; 10 mIU/ml). After 48 h, the medium (S) was removed and stored. Cells then were mechanically lysed and centrifuged at 10,000 X g. The supernatant was further centrifuged (100,000 X g) to obtain a microsomal fraction (M) and cytosol (C). The M fraction was resuspended in medium 199 with 10(-6) M pregnenolone plus 5 microM NAD+ and incubated for 2 h to determine 3 beta-o1 dehydrogenase activity. The S, C, and M fractions were all assayed for progesterone (P) by RIA. hMG markedly increased P concentrations in the S and C fractions. The M fraction demonstrated a hMG-dependent enhancement in 3 beta-HSDH activity. However, the hMG-associated S, C, and M P levels were decreased in granulosa cells coincubated with FRP. In conclusion, ovarian steroidogenesis may be dependent on the integrated interactions of both gonadotropins and local nonsteroidal paracrine/autocrine modulators of granulosa cell function.

Chlamydia trachomatis is not an important cause of abnormal postcoital tests in ovulating patients.

To examine the role of Chlamydia trachomatis infections of the cervix and abnormal postcoital tests (PCT) in a general infertility clinic, 63 consecutive patients undergoing a midcycle PCT during a routine infertility workup underwent endocervical curettage, and a 10-ml blood sample was obtained. The endocervical tissue was cultured for C. trachomatis; the serum sample was analyzed for chlamydial IgG and IgM antibodies using an indirect microimmunofluorescence assay. A negative titer was considered to be less than or equal to 1:8 dilution for IgG antibodies and less than or equal to 1:32 dilution for IgM antibodies. A good PCT was defined as greater than or equal to 5 motile sperm per high power field (HPF). A poor PCT was defined as less than 2 motile sperm/HPF, and a fair PCT was defined as 2 to 4 motile sperm/HPF. Of the 63 PCTs done, 27 (42.9%) were good, 14 (22.2%) were fair, and 22 (34.9%) were poor. All endocervical tissue cultures for C. trachomatis obtained during PCTs were negative. All IgM chlamydial antibody titers were negative (less than or equal to 1:32 dilution), 55 (87.3%) of the patients having a zero titer. Eleven (17.5%) of the patients had negative IgG chlamydial antibody titers (less than or equal to 1:8 dilution), none of the patients had a 1:16 dilution, and 52 (82.5%) had positive IgG chlamydial antibody titers (greater than or equal to 1:32 dilution). Thus, chlamydial infections of the endocervix are rare and not commonly associated with poor PCTs in this patient population.

Anticipatory grief and aged widows and widowers.

The purpose of this investigation was to explore the preventive value of anticipatory grief on the medical adjustment of the aged bereaved. Data were collected for 81 surviving widows and widowers 6 mo. after their loss. Sixteen of the bereaved had spouses who died of a chronic illness. These two groups were compared on three criteria of medical adjustment: physician office visits, feeling ill without contacting a physician, and use of psychotropic medications. The data indicated that the aged bereaved of a lengthy chronic fatal illness did worse than the bereaved of a shorter chronic illness death. By sex, no significant differences were found between widows and widowers of a short-term illness death. However, differences were noted for the chronic illness category, and within this category for length of chronicity. In both cases widowers showed more medical problems than widows.