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To obtain new criteria for presuming the existence of chromosomally unbalanced progeny in reciprocal translocation carrier couples, the obstetrical histories of 57 couples were analysed. The chromosomally unbalanced progeny who had been prenatally or postnatally diagnosed were mainly partial trisomies. In contrast with partial trisomies, there were only a few cases of partial monosomies and there were only two cases of chromosomal imbalance derived from 3:1 segregation in gametogenesis. Moreover, the percentage of haploid autosome length (%HAL) of the maternal carriers whose progeny had suffered from chromosomal imbalance was significantly smaller than those of the maternal carriers who had not had progeny with chromosomal imbalance. Based on these results, the following criteria were obtained: 1) Partial trisomies tend to be maintained in pregnancy but partial monosomies do not; Most chromosomally unbalanced zygotes which were induced by 3:1 segregated gametes tend to be spontaneously selected in early pregnancy; 3) When interchanged segments of chromosomes are small in maternal reciprocal translocation carriers, chromosomally unbalanced fetuses tend to be maintained in pregnancy.

Fluctuations in fetal hemodynamic parameters are commonly observed, and we can use the variation in the fetal heart rate as an index in fetal assessment. The purpose of this study was to clarify the spontaneous rhythm in systemic circulation in fetal lambs by means of power spectral analysis. Three pregnant sheep and their fetuses at 125 and 135 days of gestation were surgically instrumented and studied. After a minimum recovery period of four days, a fetal electrocardiogram was taken and arterial blood pressure was recorded. They were also examined in the absence of heart rate variation by means of fetal cardiac pacing. We used an autoregressive model to estimate their power spectral densities. In normal fetal lambs, two spontaneous rhythms were detected in the spectra for the fetal heart rate and arterial blood pressure. Spectral analysis of arterial blood pressure variation in the absence of heart rate variation revealed that there was only one consistent major spectral component around 0.15 Hz. This suggests that fluctuations are caused not by a variation in the heart rate but by a variation in peripheral motor activity. This spontaneous rhythm in peripheral resistance may be associated with the sinusoidal heart rate pattern in the fetal cardiotochogram.

In order to investigate the immunological mechanisms of pregnancy, fluorocytometric and immunohistochemical analysis of the cells was performed in the placenta and spleen of a murine spontaneous miscarriage model (CBA/J x DBA/2) and control (CBA/J x BALB/c). There was a significant difference between the miscarriage rate for the miscarriage model and that for the control, even though H-2 in these two group is matched. The analysis also was performed in a miscarriage model immunized with male splenocytes. Moreover, the effect of gamma-interferon, a potentiator of NK cell activity, on pregnancy was examined. Interferon treatment increased the miscarriage rate. In pregnancy, the number of splenocyte positive Asialo-GM1 or LFA-1 decreased and the intensity of these antigens decreased, as well. Interleukin-2R positive cell increased in number as well as intensity. In the miscarriage model group successfully treated by immunization, the number of Asialo-GM1 positive cells and L3T4 positive cells decreased, whereas they increased in the unsuccessfully treated group. Asialo-GM1 positive cells in the placenta of successful pregnancy decreased in number, and those in miscarried pregnancy increased. In conclusion, the success of the immunization treatment for habitual abortion depends on how to suppress NK cell activity in a linkage with the helper T-cell.

Carbamyl phosphate synthetase deficiency (CPSD) is one of the enzyme defects of the urea cycle and inherited as an autosomal recessive. A definitive enzymatic diagnosis of CPSD can be made by biochemical assay of liver biopsy material, but not of cultured fibroblasts. In pregnancy at risk for CPSD, prenatal diagnosis was attempted by fetal liver biopsy, performed at 22 weeks of gestation. CPS activity was present and a healthy baby was delivered at term. The technique employed for fetal liver biopsy is described together with an evaluation of its possible role in prenatal diagnosis.

Biophysical profile (BPP) score was assessed immediately before fetal blood sampling by cordocentesis in 150 fetuses referred to our hospital, 95 after and 55 before 30 weeks of gestation. In 95 fetuses after 30 weeks of gestation, 39 fetuses were evaluated with BPP scores of 12, 35 were from 8 to 11 and 21 were less than 7. In 55 fetuses before 30 weeks of gestation, 8 fetuses were evaluated with BPP scores of 12, 27 were from 8 to 11 and 20 were less than 7. pO2, pH and pCO2 in fetuses with a score less than 7, either before or after 30 weeks of gestation, (with a score less than 7) did not significantly differ, in comparison to the other two groups. No variables in the biophysical profile precisely reflect fetal hypoxemia, acidemia or hypercarbia. Since even the fetus with a BPP score of greater than 8 may not always be assured of well-being and not all fetuses with a score of less than 7 are necessarily in a deteriorated condition, it is necessary to evaluate fetal condition on the basis of fetal blood gas data obtained by cordocentesis, especially when the fetus is additionally handicapped by prematurity or morbidities such as growth-retardation.

One thousand forty-seven fetal samples were obtained from women who received the prenatal cytogenetic examination for different reasons. In this study, the incidence of fetal chromosomal aberration related to each reason was analyzed. The incidence of de novo chromosomal aberration in the fetuses of the higher maternal age group was 1.5% (8/525: 4 cases of 21-trisomy, 2 cases of 18-trisomy and 2 cases of 47,XXY). The incidence in the group of women who had borne (a) chromosomally abnormal child(ren) was 0.5% (1/202). There was no repeat of 21-trisomy, 18-trisomy or 13-trisomy in this study. The incidence in the group of women who had ultrasonographic abnormalities was 16.9% (26/154). In this study, abnormal amniotic fluid volume, intrauterine growth retardation, malformation, hydrops fetalis, intestinal obstruction, omphalocele, single cord artery and cerebellar hypoplasia were related to chromosomal aberrations. The incidence in the group of women whose husband or herself was cytogenetically abnormal was 40.0% (26/65). This high value indicated that parental translocation and inversion are easily transmitted to their offspring.

Cell junctions in mouse blastocyst were ultrastructurally investigated with or without lanthanum tracer. Tight junctions, gap junctions and desmosomes were observed in the trophectoderm. The tight junction was located near the zona pellucida in all trophoblast interspaces, whereas the gap junction and the desmosome, which were infrequently observed, were localized far from the zona pellucida. However, the desmosomes in the trophectoderm of the expanded blastocyst after culture increased in number and came to be located near the zona pellucida. The trophoblast layer excluded lanthanum whose invasion was interrupted by the tight junction. There were a few intermediate junctions in the interspace between the trophoblast and the inner cell mass cell as well as between the inner cell mass cells. These findings indicate that a substances whose molecular weight exceeds that of lanthanum (138.9) may not flow into the blastocyst through the intercellular space. Moreover, it is speculated that the intercellular connection is strengthened during the expansion of the blastocyst.

In order to study the morphological features of mouse embryos in the early developmental stage, we first established an in vitro culture system applying a collagen gel layer, and then observed the morphology of the embryos cultured in this system. Embryos after hatching were attached to the collagen gel layer and grew to the egg cylinder stage. By means of morphological analysis of embryos cultured for 3 or 4 days, some interesting characteristics, such as processes and villi of the mural trophoblast, lacunae formation in the mural trophoblast, steroid synthesis of the mural and polar trophoblasts and desmosome or intermediate junctions between the mural and the polar trophoblasts, were revealed. Moreover, no morphological difference was observed between cells derived from the inner cell mass. From those findings, it has been concluded that the established culture system is useful in observing the morphology of early embryonal development and also in maintaining the viability of the embryo.

The experiments were performed in order to reveal the role of prostaglandins in the implantation process, using indomethacin, a inhibitor of prostaglandin synthesis. Indomethacin was administered to mice on various days of pregnancy, and then the number of implantation sites and the uterine weight per the number of implantation sites were determined on day 10 of pregnancy. Furthermore [3H]uridine incorporation of blastocysts incubated in the medium containing indomethacin with/without prostaglandin F2 alpha was evaluated in vitro. The results were as follows. Indomethacin reduced the number of implantation sites in the uterus and the uterine weight per the number of implantation sites (embryonal growth). Indomethacin inhibited the [3H]uridine incorporation of blastocysts, but prostaglandin F2 alpha counteracted the effect of indomethacin. These results suggested that prostaglandins play a role in inducing implantation and embryonal growth, and that blastocysts might synthesize prostaglandins.

With flow-cytometry we tested whether or not interferon can induce surface expression of HLA-ABC antigen on cell lines derived from various gynecologic cancers. The tumor cell lines used were Hela S3 derived from cervical cancer, OVK-18 derived from ovarian cancer, HHUA derived from endometrial cancer, SCH, JaR and BeWo derived from choriocarcinoma. 1000 IU/ml of interferon-gamma induced HLA-ABC antigen expression on Hela S3, OVK-18 and SCH but not on BeWo. HLA-ABC antigen was expressed on neither the surface of HHUA or JaR nor was its appearance induced by interferon. The clinical application of interferon may be effective in killing some tumors, taking advantage of the increased expression of HLA antigen.

Ascitic fluid was obtained from the patient of ovarian endometrioid cancer. Collected cells were incubated with Eagle's MEM containing 15% fetal calf serum at 37 degrees C under humidified 5% CO2 and 95% air. The epithelial colonies grew rapidly and were released without fibroblast cells. After the first passage, the cells are growing without interruption for over one year and 35 transfer generations. This cell line has following charactors: 1) The monolayer cultured cells appeared epithelial, pavement like arrangement and piling up, without contact inhibition. 2) In the cytoplasma PAS positive substance can be seen. 3) Desmosome-like structure, gap junction, microvilli and well developed cell organelles can be found by electron microscopy. 4) Chromosomal number shows pseudodiploidy which mode is 47. A submetacentric chromosome was present in all karyotype and identified by G-banding. It consists of No. 11 and a part of No. 1. 5) By heterotransplantation to the nude athymic mouse, the tumor easily develops. 6) By estradiol and promegestone its growth was inhibited. 7) Estrogen and progesterone receptors were not detected in the cytosol.

Mouse blastocysts collected on day 4 were cultured for 20 hours in media containing various concentrations of [3H]estradiol with/without 10(-6) M unlabelled estradiol. There was a significant reduction in the radioactivity of embryos cultured at 4 degrees C compared with that of embryos cultured at 37 degrees C. The radioactivity of embryos kept in the washing solution after washing was slight lower but showed no significant difference when it was compared with that of embryos not kept in this way. There was no significant difference between the radioactivity of embryos cultured in 2 X 10(-8) M [3H]estradiol and that of embryos cultured in 2 X 10(-8)M [3H]estradiol plus 10(-6)M non-labelled estradiol. Furthermore the radioactivity of embryos incubated in 2.5 X 10(-8)M [3H]estradiol is more than twice that of embryos incubated in 10(-8)M [3H]estradiol. These results demonstrate that preimplantation mouse embryos are capable of uptake and retaining of estrogen and that the mechanism of uptake is neither selective nor receptor mediated.

Human granulosa-luteal cells obtained from dominant preovulatory follicles at the time of oocyte retrieval for in vitro fertilization (about 10 h after the peak of the endogenous LH surge) produced large quantities of progesterone (P) (8.7 pg/cell/first 2 days) in a monolayer culture. Although basal production of estradiol (E2) in these granulosa-luteal cells was restricted to 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 in these granulosa-luteal cells and it did not inhibit T aromatization. In contrast, DHT did not increase E2 production in cultured granulosa cells obtained from follicles 2 approximately 5 days before the LH surge. Thus, the response of granulosa cells to exogenous androgen seems to vary at different stages of cell differentiation. Neither T nor DHT elicited significant changes in the production of P by granulosa-luteal cells obtained from preovulatory follicles during the LH surge. However, T (10(-6) M) suppressed hCG-stimulated P production in these cells. The results suggest that androgen enhances the estrogen biosynthesis of granulosa-luteal cells obtained from preovulatory follicles during the LH surge not only by acting as substrate for aromatization but also by participating in some process of estrogen synthesis. Further, it is suggested that androgen has a negative effect on P biosynthesis in these cells.

We have evaluated serum estradiol, progesterone, testosterone, and urinary estrogen excretion in 24 hour urine samples to monitor indices of follicular maturation. The serum steroid levels were determined with the direct radioimmunoassay kit. The urinary estrogen level was measured with the estrogen micrometering kit using hemagglutination inhibition reaction. Moreover, relationships between these steroid levels and the follicular size measured with ultrasound were analyzed. The serum estradiol concentration and the estrogen excretion in 24 hour urine samples mostly showed a continuous increase during the late follicular phase, and had a positive correlation to the maximum follicular diameter of the leading follicle (MxFD) and to the total of the maximum diameter of the follicles (TFD). The serum progesterone concentration showed a remarkable increase especially on the day of the LH surge onset in many cases, and had a significant (p less than 0.01) correlation to MxFD but not to TFD. The serum testosterone concentration, however, showed neither a specific tendency on its daily change nor a correlation with the follicular size. These results indicated that the serum estradiol and progesterone, and the urinary estrogen excretion can be utilized as indices of follicular maturation.