Toshio Nishimura

Progesterone-induced smooth muscle—like cells in the subperitoneal nodules produced by estrogen: Experimental approach to leiomyomatosis peritonealis disseminata

In order to examine the pathogenesis and histogenesis of leiomyomatosis peritonealis disseminata (LPD), guinea pigs were treated with various doses of estradiol benzoate (E) (40, 80, 100, and 200 microgram/day, three times a week, by intramuscular injection). After treatment for 3 months, each dose of E produced lesions simulating the disseminated pattern of LPD. Ultrastructurally, the nodules were considered to be composed of cells resembling those of fibroblast. In guinea pigs, pretreatment with estradiol benzoate (100 microgram/day, three times a week intramuscularly for 3 months followed by combination treatment with E and progesterone (P) (0.5 and 1.0 mg/day, once a week, intramuscularly) for 1 to 3 months produced nodules similar to those with estrogen alone. Ultrastructurally, these nodules were composed of cells resembling smooth muscle and decidual cells. According to these results, we conjectured that estrogen differentiates and proliferates cells with the features of fibroblast-like cells from the subcoelomic totipotential mesenchyme, and these cells are differentiated into smooth muscle-like cells and decidua-like cells under the influence of both estrogen and progesterone, resulting in the production of multiple subperitoneal nodules.

Possible presence of autoantibodies to zona pellucida in infertile women

Of 52 serum samples from infertile women tested against isolated porcine oocytes by immunofluorescence, 8 produced intense reactions in zonae pellucidae, while only one of 52 sera from control subjects showed the reaction. Autoantibodies to the zona may be present in these women.

Radioimmunoassay of maternal serum α-fetoprotein associated with pregnancy

n Alpha fetoprotein (AFP) was measured by radioimmunoassay of urine and serum of pregnant women at various stages of gestation and postpartum; sera of patients with hydatidiform mole and choriocarcinoma were also measured. Concentrations below 5 ng/ml were considered negative. Nonpregnant and early pregnancy levels of AFP did not differ significantly. At 14 weeks of gestation, serum AFP showed an initial increase which increased progressively throughout pregnancy, reaching its maximum 35-38 weeks postconception. Average serum AFP was 232.6 ng/ml at 35 weeks, 253.3 at 36 weeks, 222.3 at 37 weeks, and 185.7 at 38 weeks. At term, serum AFP leveled off slightly, averaging 108.7 ng/ml at 40 weeks, 86.5 at 41 weeks, and 80.8 at 42 weeks. Serum AFP decreased rapidly after delivery and was almost negligible by 20 days postpartum. Average AFP half-life was 3.9 days. AFP was also detected in serum from 1 patient with hydatidiform mole (105 ng). AFP was found in urine of 1 patient with marked albuminuria but not in normal pregnancy. Results indicated that AFP in serum may be partly of maternal origin. It is speculated that the inhibition against the repressor-operator gene system which was responsible for AFP synthesis was accelerated in pregnancy by unknown factors, probably of placental origin, resulting from maternal AFP production.n

A new method for preparation of pure zonae pellucidae in large quantities from porcine ovaries

A new method for preparation of large quantities of zona substance in pure form from porcine ovaries has been described. In addition to the previously reported techniques of glass wool treatment and sieving by saran meshes, the following three significant improvements have been introduced: disruption of ovaries by an electric machine equipped with two multi-needed disks, which resulted in a considerably accelerated recovery of follicular oocytes with a high yield; low-speed centrifugation at 170 X g for 15 s was found to be an obligatory step to eliminate light particulate material from the crude oocyte suspension; use of 50% sucrose solution in discontinuous density gradient centrifugation permitted complete separation of homogeneous samples at two stages of the final preparation of zona-encased oocytes or of oocyte-free zonae. Microscopic examination revealed no contaminating components in the zona preparation. With this method 93 mg of lyophilized zona preparation were obtained from 24 553 porcine ovaries. Analysis of a solubilized zona substance by Sephacryl S-200 column chromatography showed the presence of two major glycoproteins which could not be separated completely from each other. By analysis of the two components with SDS-PAGE, only a single, but broad, band of glycoprotein was found, indicating the successful isolation of a major component(s) from porcine zonae.

Autoantibodies to zona pellucida in infertile and aged women.

A total of 735 human serum samples were tested against isolated porcine ova for detection of autoantibodies to zonae pellucidae by the specific method reported previously using indirect immunofluorescence. Positive fluorescence was found in 13 of 175 samples (7.4%) from patients with unexplained infertility, in 2 of 122 samples (1.6%) from patients with proven etiology of infertility, but in none of 141 samples from age-matched control subjects (fertile women and men). There was an age difference in the incidence of positive reactions: 4 of 156 samples (2.6%) from women over 40 years old were positive in contrast to only 1 of 159 samples (0.6%) from women under 40 years old. Although autoantibodies to zonae pellucidae may be produced during the aging process, the observed high incidence in unexplained infertility suggests an etiologic association of the antibody with the disease when produced in younger women.

Significance of atretic follicles as the site of androgen production in polycystic ovaries

To examine androgenic potential of polycystic ovaries (PCO), slices of follicular and stromal tissues from the same ovary obtained by wedge resection from two markedly, two moderately and one slightly enlarged PCO were incubated separately with [1-14C] acetate. Incorporation into progestins, androgens and estrogens was assessed by reverse dilution technique with recrystallization to constant specific activity. Although the greatest incorporation into androstenedione with much lesser incorporation into testosterone and dehydroepiandros-terone was observed with all the five follicles, the amount of incorporation into the three steroids increased gradedly with histologically defined magnitude of thecal cell hyperplasia in atretic follicles. Only the stromal tissues from two markedly enlarged PCO produced androgens with a similar pattern of 14C distribution among the steroids, thereby incorporation into the three androgens remaining 5.5% or less of that by the follicles from the same ovary. Preoperative levels of plasma androstenedione and urinary 17-ketosteroids were shown to increase in four patients with PCO containing atretic follicles with thecal cell hyperplasia, but not in one patient with slightly enlarged PCO containing atretic follicles without thecal cell hyperplasia. It is inferred that atretic follicles with thecal cell hyperplasia is a significant source of androgen overproduction by PCO.

Suppression of mixed lymphocyte reaction by progesterone and estradiol-17β

The effects of estrone, estradiol-17beta, estriol, testosterone, progesterone, and 20alpha-dihydroprogesterone on incorporation of 3H-thymidine into cultured lymphocytes from normal men and nonpregnant women in response to allogeneic stimulation was investigated with two-way MLR. A dose-dependent reduction in the mean percent incorporation was observed in the range of progesterone concentration between 2 x 10(3) and 10(4) ng/ml with statistically significant differences from the control cultures. Estradiol-17 beta at 10(4) ng/ml produced a slight but significant inhibition of incorporation. No significant change in incorporation occurred with the other steroids tested. Since the suppressive property of progesterone in phytohemagglutinin-induced lymphocyte culture reported in our previous communication was reproduced by the present investigation in MLR, the possible role of progesterone in immunoregulation during pregnancy is emphasized as has been postulated previously.

Electrogenic Absorption of Sugars and Amino Acids in the Small Intestine of the Human Fetus

Some characteristics and the degree of intestinal absorption in the developing human fetus were examined by measuring solute evoked potentials and 14C-D-glucose uptake into the everted jejunal segments. In all segments, the Michaelis-Menten relationship was observed between D-glucose concentrations and the evoked potentials or D-glucose uptake. Increase of Na-ion concentrations enhanced both D-glucose evoked potentials and uptake. Both D-glucose and L-alpha-alanine evoked potentials increased in a significant correlation to the fetal age; however, the apparent Michaelis constants did not show any signficant change. The structural specificity of sugar for generating evoked potentials was the same as that reported for adult mammals. Among amino acids, only the L-form of neutral and acidic amino acids generated markedly high evoked potentials, but the basic ones hardly at all. Oligopeptides such as glycyl-glycine and glycyl-glycyl-glycine also generated high evoked potentials. Our results have indicated that the active transport system of sugars and amino acids in the human fetus have already developed by as early as the sixth month of gestation.

Oxytocin sensitivity and effects of estrogen and progesterone on metreurynter-induced abortions at midpregnancy: Preliminary report

Abstract The relationship between mechanical stimulation given to the uterus and the resulting alteration in uterine contractility was studied; the effects of estrogen and progesterone given by different routes on the above relationship were also assessed. A metreurynter was inserted in the uterus of 51 patients at midpregnancy, and the time until positive oxytocin sensitivity was studied. Stimulation by metreurynter universally elevated the oxytocin sensitivity of the uterus, but without the help of a metreurynter, no elevation of oxytocin sensitivity was brought about. As to the effect of the gonadal hormones given on this process, no definite conclusion was drawn.

Radioautographic demonstration of dihydrotestosterone receptor in cultured human fibroblasts

Radioautographically, we obtained direct evidence for the localization of3H-dihydrotestosterone binding sites in cultured human fibroblasts derived from vulvar skin.