Search

We investigated whether growth hormone(GH) may influence the development of mouse embryo in vitro using recombinant GH and anti-growth hormone receptor(GHR) antibody. When mouse two-cell-stage embryos were cultured with GH, 5, 10 mIU/ml, the rates of formation of both blastocysts and of hatched blastocysts from two-cell-stage embryos were significantly higher than in the medium without GH. The number of blastomeres from blastocyst treated with GH, 2, 5 and 10 mIU/ml, was significantly higher than those cultured without GH. When cultured blastocysts were implanted in recipient uteri, a significantly higher implantation rate was found for the GH-treated(10 mIU/ml) mouse embryos than for controls. To confirm the GH effect, anti-GHR monoclonal antibody MAb5 was employed to neutralize the action of GH through GHR. Two-cell-stage mouse embryos were cultured with GH, 10 mIU/ml, and anti-GHR antibody, and as a control, embryos were cultured with GH alone. Both the rate of formation of blastocysts from two-cell-stage embryos and the number of blastomeres were significantly decreased in groups exposed to anti-GHR antibody. Results indicate that GH may be important to early embryonic development, acting through GHR. GH may help to obtain good quality embryos and improve the implantation rate in IVF programs.

In vitro co-culture of embryos and somatic cells is used to obtain well-developed embryos of humans and other species. However, it is not known whether direct cell-to-cell contact is essential to embryonic development. To elucidate this mechanism, we cultured mouse embryos using rabbit oviductal cell as a somatic cell. To avoid the direct contact we used a microporous membrane cell-culture insert. This cell-culture insert permits only the liquid portion of the culture medium to pass through it and was interposed between mouse embryos and cultured rabbit oviductal cells. When mouse embryos were placed on the cultured oviductal cells directly, 52.2% of two-cell-stage embryos developed to the blastocyst stage. In contrast, when the cell-culture insert was interposed between the embryos and the cultured oviductal cells, 50.0% of two-cell-stage embryos developed to that stage. There was no difference between the rate of blastocyst development in co-culture system with and without cell culture insert. Results indicate that oviductal factor (s), rather than a direct contact with oviductal cells, is essential to the enhancement of embryonic development in vitro.

To investigate the criteria for laparoscopic surgery in ovarian tumors, a retrospective review of indications, preoperative assessments using transvaginal ultrasonography, serum CA 125, surgical methods and diagnosis and post-operative complications were analyzed. Forty-three patients underwent surgical treatment for ovarian tumors, from November 1992 to October 1993. Of the 43 patients with ovarian tumors, 19 (44%) underwent laparoscopic surgery, and 24 (56%) underwent laparotomy. Laparoscopic surgery was performed in 18 of 34 (53%) patients with benign ovarian cysts. Eight patients with malignant ovarian tumors were treated by laparotomy. Coincidence of preoperative assessment and surgical diagnosis was 95%. No major complications were observed in the laparoscopic surgery. We suggest that malignant ovarian tumors and inextricable adhesions are contraindications to laparoscopic surgery. We propose the following criteria for laparoscopic surgery by classification into three groups: A) good indication: benign cystic tumors with good mobility, B) deliberation indication: benign tumors with poor mobility, including some dermoid cysts and endometrial cysts, C) contraindication: malignant tumors and dense adhesions. We conclude that indications of laparoscopic surgery for ovarian tumors should be determined by careful preoperative assessment using transvaginal ultrasonography and tumor markers.

The clinical and pathological features of an apparently unique case of an endometrial cyst of the uterus are reported. The cyst was located within the myometrium of a 16-year-old woman suffering from dysmenorrhea. After excision of the cyst, patient's symptoms improved. On histological examination, the cyst most closely resembled an adenomyotic cyst.

What parameter of semen has influence on the pregnancy outcome of artificial insemination with the husband's semen (AIH) with washed sperm for male infertility was investigated. Two hundred fifteen cycles in 62 patients of AIH with washed sperm for male infertility were prospectively studied. We compared six parameters (total sperm count, motility rate, sperm motile efficiency (SME) both before and after washing) in pregnant and non-pregnant cycles. SME of washed sperm was significantly higher in the pregnant than in the non-pregnant cycles (241.0 +/- 68.0 vs. 169.5 +/- 80.0). There were no differences in other five parameters between pregnant and non pregnant cycles. SME, the parameter of sperm motility characteristics, of washed sperm is closely related to the outcome of AIH for male infertility. Sperm motility characteristics after washing may be one of the important indexes of the result of AIH for male infertility.

Macrophage colony stimulating factor (M-CSF) concentrations in the peritoneal fluid of 44 infertile patients were measured. The concentration of M-CSF was significantly higher in the patients with endometriosis than in those without endometriosis. There was no significant difference between the M-CSF concentration and stage of endometriosis classified by revised American Fertility Society (rAFS) classification in the 44 patients. However, in primary infertility patients with endometriosis there was a significant correlation between peritoneal M-CSF levels and rAFS points of endometriosis. We speculate that there is some relationship between the increased levels of M-CSF in the peritoneal cavity and the primary infertility associated with endometriosis especially progression of endometriosis.

The efficacy of long term consecutive treatment of clomiphene citrate (CC) was investigated in 608 infertile women associated with anovulatory disorders (classified as WHO group II amenorrhea). This study was performed in multi-centric facilities of obstetrics and gynecology by a retrospective randomized assessment. Infertile women associated with anovulation were treated by 50-150 mg of CC for five consecutive days in each treatment cycle. In this study, 200 pregnancies were observed and the total pregnancy rate was 32.9%. The number of abortions were 33 out of the 200 pregnancies (16.5%). The cumulative pregnancy rate within the pregnant subjects reached 90% in initial 10 treatment cycles and there was no difference between 50 mg and 100 mg of CC treatment. There were remarkable differences among the pregnancy rates achieved during consecutive treatments at the primary (clinics and hospital), secondary (regional center hospitals) and advanced facilities. Consecutive treatment by CC for infertile women associated with anovulation may be effective during initial 10 cycles. Although ovulation can be confirmed during the following cycles, the stimulation protocol should be reconsidered after 12 consecutive cycles therapy with CC, because cycle fecundity decreases. Hence, to improve pregnancy rates, it is important to check the previous administration of CC which reflects adversely on the subsequent conception rate especially in advanced facilities where these patients are referred to.

Although an increased incidence of shared HLA antigens among couples experiencing recurrent spontaneous abortion or infertility has been observed, antigenic effects remain to be clarified. These effects, which are suspected to exist among fertile couples, were examined in this study. A group of 247 healthy Japanese families and their 682 children, whose HLA antigen haplotypes could be determined, were studied. Haplotypes of the fathers were compared with those of the mothers, and the combination of HLA antigens in the A, B, C and DR loci was determined as antigenic patterns against the mother. Patterns inherited by children were examined and deviations of the patterns were studied according to the natural rule that either of the haplotypes should be inherited at equal rates. Lower rates of children with antigens common with those of the mother, which may show a result of reproductive selection, were observed among only the first born children in the A (p < 0.05) and DR loci (p < 0.01), but rates similar to those expected were observed among the second born and later children. Lower rates were also observed in the B and C loci among all children with no significant difference between groups of the children. The necessity of differing antigenic factors in order to achieve pregnancy in the population was shown. Important effects of HLA antigenicity in reproductive failure are suggested.

The possible physiological significance of endogenous inhibin was evaluated in prepubertal female rats during sexual maturation. Ovarian/serum inhibin, serum FSH levels were measured from 4th day to 35th day of life by RIA. Serum inhibin levels were first detected on 4th day of life, thereafter, progressively increasing up to 27th day. Serum FSH levels gradually increased up to the 14th day with a sharp fall in the FSH levels on day 15 which afterwards was maintained at a lower level. Pregnant mare serum gonadotropin (PMSG) treatment on day 14 and day 22 rats showed different patterns of serum FSH and inhibin changes. In 14 day group, there was no difference between control and PMSG treatment, but in 22 day group, not only significant difference between control and PMSG treatment but also inverted relations between FSH and inhibin after PMSG treatment were found. It can be postulated from these findings that secretion of inhibin from the granulosa cells is initiated by FSH by its direct action on the ovary whereas the level of FSH itself is controlled by inhibin after FSH reaches peak values. Also it can be inferred that the feedback control mechanism of inhibin seems to be established around 3rd week of life.

We measured maternal serum unconjugated estriol (SUE3) levels in 244 cases (218 cases with labor and 26 without labor) and investigated the relationships among several obstetrical factors and SUE3 levels. There was no significant difference in the SUE3 level between the group with labor and the group without labor. However, the SUE3 level decreased gradually with prolonged duration of labor in multipara. There were significant positive correlations between the SUE3 levels and birth weights (n = 82, r = 0.375, p less than 0.01) or placental weights (n = 82, r = 0.381, p less than 0.01) in the multipara with labor. The SUE3 levels of fetal distress cases (n = 30, 20.0 +/- 8.6 ng/ml) were significantly (p less than 0.05) lower than those without fetal distress (n = 188, 24.5 +/- 9.7 ng/ml). These data suggest that maternal SUE3 might have been affected with feto-placental function, even after the onset of labor SUE3 levels decrease with prolonged duration of labor.

Adrenocorticotropic hormone (ACTH), beta-lipotropin (beta-LPH) and beta-endorphin (beta-EP) immunoreactivities were measured by high sensitive radioimmunoassay in maternal and umbilical cord plasma samples which were obtained simultaneously in 12 cases. Mean ACTH levels in cord and maternal plasma were significantly higher than those of 8 normal adults. Mean beta-EP to beta-LPH molar ratio of 0.19 in maternal plasma was different from that of cord plasma (0.13). There was no significant correlation between beta-EP levels, or between beta-LPH levels, in the paired fetal and maternal samples. These data suggest that beta-LPH and beta-EP elevate during labor and delivery in response to the stress. Beta-LPH and beta-EP in cord plasma are of fetal and/or placental origin.

Pregnant Wistar rats were daily injected with 5 mg sulpiride into the peritoneal cavity starting on day 14 of gestation. Blood samples were obtained from the maternal carotid artery and fetal axillary vessels on day 20 of gestation. Serum prolactin (PRL) levels were determined by radioimmunoassay using a rat PRL RIA kit. The maternal serum and fetal plasma PRL levels were significantly higher in the sulpiride-treated group than in the saline control group. There was a significant positive correlation between fetal body weight and fetal plasma PRL level. These results suggest that sulpiride which reached the fetus stimulates fetal PRL secretion and that PRL may exert a growth-promoting effect on the fetus.

Prolactin in the amniotic fluid is known to have an osmoregulatory function in the fetus, but little is known of the effects prolactin in the exudate from the endometrium acting directly on the early embryo. ICR female mice were stimulated with 6 IU of PMSG (pregnant mare serum gonadotropin) and 8 IU of hCG (human chorionic gonadotropin) and mated with ICR males. Two-cell embryos through blastocysts were obtained for in vitro culture, to which prolactin was added at doses of 0, 10, 30, 100, 300 or 1000 ng/ml. The effects of different doses of prolactin were compared for the rates of blastocyst formation and hatching, and for the development of the embryo with regards to the stage of embryo from which the culture was launched. The results show that prolactin promotes development of the two-cell embryo at concentrations of 300 ng/ml and above, improving the rate of formation of the blastocyst and hatching stages. It also has positive effects on post-hatching blastocyst cultures, in terms of acceleration in development.

We measured beta-endorphin (beta-EP) and beta-lipotropin (beta-LPH) levels in human maternal and fetal plasma and amniotic fluid, simultaneously. It appeared evident that maternal circulating levels of beta-EP (n = 11, 163.9 +/- 12.9 pg/ml, mean +/- S.E.) and beta-LPH (n = 11, 413.0 +/- 25.9 pg/ml) at delivery were significantly (p less than 0.01) higher than those of maternal plasma at term (beta-EP; n = 4, 18.3 +/- 2.1 pg/ml, beta-LPH; 213.4 +/- 24.3 pg/ml) and those of amniotic fluid (beta-EP; n = 5, 8.5 +/- 1.2 pg/ml, beta-LPH; 215.1 +/- 44.9 pg/ml). Fetal beta-EP levels (n = 11, 79.1 +/- 5.8 pg/ml) were significantly (p less than 0.01) higher than those of amniotic fluid. These data suggest that the origin of amniotic fluid beta-EP may be an increased synthesis in the maternal and fetal pituitary gland but not in the placenta.

Serum prolactin and estradiol-17 beta levels were determined in umbilical cord sera obtained from 33 preterm, 118 full-term and 11 anencephalic infants and in peripheral sera obtained from 5 neonates. Prolactin levels increased from mid-gestation toward term, and markedly decreased after birth. Those in anencephalic infants (253 +/- 86 ng/ml, mean +/- S.D.) were lower than in full-term infants (324 +/- 119 ng/ml), but the difference was not statistically significant. We also performed thyroid-releasing-hormone (TRH) (50-200 micrograms) test on 5 anencephalic infants, but significant changes of serum prolactin levels were not found. Estradiol-17 beta levels in anencephalic infants were significantly lower than those in full-term infants. There was a significant positive correlation between cord prolactin and estradiol-17 beta levels in full-term infants. These results suggest one of the control factors for human fetal pituitary prolactin secretion is estrogen, and hypothalamic factors are not so important.

We measured maternal venous (MV), umbilical arterial (UA) and umbilical venous (UV) cortisol levels in 180 term pregnancies to investigate the changes of these hormone levels in the cases of fetal distress and neonatal asphyxia. The cortisol levels in UA and UV blood in the fetal distress group were significantly higher than those in the non-fetal distress group among spontaneous vaginal delivery cases. There were significant positive correlations among the cortisol levels in MV and UA, MV and UV, and UA and UV blood, respectively. These data suggest that in the case of fetal distress or neonatal asphyxia, mother and fetus have responded independently to the stress of delivery, and that maternal cortisol might not strongly affect the fetal cortisol secretion.

In order to determine whether androgen acts solely as a substrate for aromatization or whether it also influences on the activity of aromatase enzyme, human granulosa cells were incubated in vitro with or without androgen. Although basal production of 17 beta-estradiol (E2) in cultured granulosa cells obtained from follicles about 26 hr after the initiation of the LH surge was restricted in small quantities, a marked increase in E2 production occurred in the presence of testosterone (T) (10(-6) M) as aromatizable substrate. The non-aromatizable androgen, 5 alpha-dihydrotestosterone (DHT) (10(-7) M, 10(-6) M), slightly enhanced E2 production and it did not inhibit T aromatization in these cells. By contrast, DHT did not increase E2 production in granulosa cells obtained from follicles 2-5 days before the LH surge. The results indicate that androgen can enhance the estrogen biosynthesis of granulosa cells obtained from follicles during the LH surge not only by acting as a substrate for aromatization but also by participating in some process of the estrogen synthesis. This effect of androgen was not clearly seen in granulosa cells obtained before the LH surge, suggesting that the response of granulosa cells to exogenous androgen varies with the stage of the cell differentiation.

We measured the serum cortisol levels in 15 normal and 8 pregnancy induced hypertension (PIH) primigravidas. The normal pregnancy had a significant (p less than 0.05) higher cortisol level than that of PIH patient. Especially, 4 severe PIH patients had a significantly (p less than 0.05) lower cortisol level. The lower maternal cortisol level of PIH patients became more significant with the severity of clinical symptoms of PIH. After the onset of labor, the cortisol levels of PIH patients did not elevate compared with normal pregnancy. These data suggest that in the PIH patient the reactivity of the adrenal cortex to adrenocorticotropic hormone (ACTH) may be blunted or the cortisol production in the adrenal cortex may be decreased by the tissue circulation insufficiency due to PIH.

Growth hormone (GH) and prolactin (PRL) in maternal and fetal serum and amniotic fluid were measured simultaneously by radioimmunoassay at term deliveries of 20 normal mothers. The PRL level of amniotic fluid was significantly higher than that of maternal and fetal serum. The fetal GH level was significantly higher than the maternal and amniotic levels of GH. There was no significant correlation among hormone levels of maternal and fetal sera and amniotic fluid. These data suggest that PRL and GH levels of maternal and fetal sera and amniotic fluid are controlled separately.