Hiroshi Hoshiai

Inhibin: a new circulating marker of hydatidiform mole?

OBJECTIVE--To define the concentrations of inhibin in serum and tissue of patients with hydatidiform mole and assess their value as a clinical marker of the condition. DESIGN--Prospective study of new patients with hydatidiform mole, comparison of paired observations, and case-control analysis. SETTING--A university hospital, two large public hospitals, and a private womens clinic in Japan. PATIENTS--Seven consecutive referred patients seen over four months with newly diagnosed complete hydatidiform mole, including one in whom the mole was accompanied by viable twin fetuses (case excluded from statistical analysis because of unique clinical features). All patients followed up for six months after evacuation of molar tissue. END POINT--Correlation of serum inhibin concentrations with trophoblastic disease. MEASUREMENTS AND MAIN RESULTS--Serum concentrations of inhibin, human chorionic gonadotrophin, and follicle stimulating hormone were compared before and seven to 10 days after evacuation of the mole. Before evacuation the serum inhibin concentrations (median 8.3 U/ml; 95% confidence interval 2.4 to 34.5) were significantly greater than in 21 normal women at the same stage of pregnancy (2.8 U/ml; 2.1 to 3.6), and inhibin in molar tissue was also present in high concentrations (578 U/ml cytosol; 158 to 1162). Seven to 10 days after evacuation inhibin concentrations in serum samples from the same patients declined significantly to values (0.4 U/ml; 0.1 to 1.4) similar to those seen in the follicular phase of normal menstrual cycles. None of the four patients whose serum inhibin concentrations were 0.4 U/ml or less after evacuation developed persistent trophoblastic disease. Though serum human chorionic gonadotrophin concentrations declined after evacuation (6.6 x 10(3) IU/l; 0.8 x 10(3) to 32.6 x 10(3], they remained far higher than in non-pregnant women. Serum follicle stimulating hormone concentrations remained suppressed. CONCLUSIONS--In this small study serum inhibin concentrations higher than those found in the early follicular phase one to two weeks after evacuation of a hydatidiform mole seemed to be specific for persistent trophoblastic disease. Further data are needed to confirm these promising results.

Effects of indomethacin, prostaglandin E2, prostaglandin F2α and 6-keto-prostaglandin F1α on hatching of mouse blastocysts

Abstract The present study has been performed to investigate how PGs would participate the hatching process. Effects of indomethacin, an antagonist to PGs biosynthesis, on the hatching of mouse blastocysts were examined in vitro. Furthermore, it was studied that prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) or 6-keto-prostaglandin F1α (6-keto-PGF1α) were added to the culture media with indomethacin. (1) The hatching was inhibited by indomethacin yet the inhibition was reversible. (2) In the groups with indomethacin and PGE2, no improvement was seen in the inhibition of hatching and the inhibition was irreversible. (3) In the groups with indomethacin and PGF2α, inhibition of hatching was improved in comparison with the group with indomethacin. (4) In the groups with indomethacin and 6-keto-PGF1α, no improvement was seen. The above results indicated that PGF2α possibly had an accelerating effect on hatching and a high concentration of PGE2 would exert cytotoxic effect on blastocysts.

Sperm-Egg Interactions Observed by Scanning Electron Microscopy

Successful in vitro fertilization requires mature oocytes in which the first polar body has been extruded and capacitated sperm capable of penetrating the zona pellucida. In this study we made a time sequential observations on human sperm-egg interactions by SEM in two experimental systems. Human sperm-human zona pellucida interaction: Cytoplasmic processes of corona cell extend around sperm head. Spermatozoa took different angles in attaching or penetrating to the zona pellucida. The head of some spermatozoa bound to the zona were vesiculated, suggesting the progression of the acrosomal reaction. Initially, the anterior part of the sperm head penetrates from the pore of the zona pellucida. Human sperm-zona-free hamster egg interaction: Most spermatozoa lie flat on the vitellus surface covered with numerous microvilli, but a few are oriented perpendicular to the vitellus surface. Most bound sperm had lost their acrosomal caps, because a ridge exists at the leading edge of the equatorial segment. Initially most microvilli appeared to grasp and immobilize the anterior tip of the sperm head. But as gamete interaction proceeded, microvilli were overlying the postacrosomal region and were observed adjacent to the plasma membrane of the postacrosomal region. The postacrosomal region is first incorporated into the ooplasma, the anterior tip of sperm head being the last portion to be incorporated. The microvilli of the oolemmal surface where sperm penetrated did not show major changes in size or in appearance, and the so-called incorporation cone was not observed.

Establishment and characterization of ovarian endometrioid carcinoma cell line

Ascitic fluid was obtained from a patient with ovarian endometrioid carcinoma. The cells obtained from the fluid were cultured in Eagles minimum essential medium containing 15% fetal calf serum at 37 degrees C under humidified 5% CO2 and 95% air. Epithelial colonies grew rapidly in the primary culture and we were able to remove them from the dish without contamination of fibroblastic cells. In the subculturing process, the cells were allowed to proliferate for over 2 years, developing 40 transfer generations. The morphology, chromosomes, and hormonal responses of the ensuing cell line were then documented.

Measurement of urinary estrogen level with the urinary estrogen micrometry method in in vitro fertilization and embryo transfer therapy

We have evaluated a new micrometering kit using hemagglutination inhibition reaction for determination of total estrogens in urine of normal fertile women and women undergoing in vitro fertilization and embryo transfer (IVF-ET). The micrometering kit was sensitive enough to monitor follicular maturation, and its technique was simple and rapid. Results were obtained in 2.5 hours. By the analysis of the individual estrogen levels in urine collected every 4 hours from 20 IVF-ET patients, the following results were found, with a few exceptions: once the level had exceeded 60 ng/ml, the onset of the luteinizing hormone surge was induced within the next 72 hours; when the level had been under 60 ng/ml, the onset was not induced within the next 24 hours

Monitoring follicular maturation through measurement of urinary estrogen excretion by latex agglutination inhibition reaction.

We evaluated the clinical utility of the MS 8601 kit, which is a new method of measurement of estrogens in urine. Changes in daily urinary estrogen excretion measured with the kit were highly correlated with that in daily serum E2. This kit was sensitive enough to detect the rises in estrogens in the follicular phase not only in women with normal menstrual cycle, but also in hMG-hCG treated patients.

In vitro fertilization and embryo transfer at Tohoku University, Sendai, Japan

100 mg/day of clomiphene-citrate is administered for 5 days from the fifth day of the menstrual cycle. Ultrasonography and highly sensitive immunoassay of urinary estrogen (Mochida Co., Japan) are used to decide the beginning of urinary LH measurement every 4 hr. We do laparoscopic egg collection 2628 hr after the onset of LH surge or 36-37 hr after HCG injection, depending on the maximum follicular diameter and urinary LH level measured with Hi-Gonavis (Mochida Co.). Incubation is done with Wittinghams T 6 medium including 10% (IM) or 15% (GM) inactivated patients serum under conditions of mixed gas (5% CO2, 5% 02, 90% N2). Six hours after preincubation of the ovum, 3-5 • 10~/ mt of washed sperm is added and incubated for 16 hr of fertilization time. Then the embryo is moved to growing medium and cultured for 36 hr. Fifty-six hours after egg recovery, the embryo is replaced into the uterus with Monash ET Catheter Type 2. Table I. Clinical Results of IVF and ET (February-September 1983, Tohoku University, Sendai)

Morphologic Differentiation of Human Immature Teratoma Cell in vitro

Immature teratoma cells obtained from a patients ovary were cultured and morphologically characterized in the early culturing stage. The major part of the tumor was composed of neuroglial-like tissues which were surrounded by neuroepithelial-like tissues. The neuroglial-like cells cultured in vitro morphologically differentiated to the epithelioid cells and the morphologic pattern of the culture came to resemble that of the original lesion. Furthermore, some kinds of the differentiated cell grew out from the cell aggregate that grew on the colonies of the neuroglial-like cells. The morphological character of the cultured immature teratoma cells suggests that neuroglial-like cells have totipotency to change to differentiated cells forming a teratoma.

Laparoscopic evaluation of the onset and progression of endometriosis

the time of catheter removal. Median follow-up for the IP cisplatin and IP mitoxantrone group has been 24 and 30 months, respectively. Ten of the 41 patients (24%) who received IP cisplatin and 4/15(26X) of the IP mitoxantrone patients have recurred (P = ns). Median time to recurrence was 18 months for both groups. This study indicates that consolidation intraperitoneal chemotherapy following negative second-look laparotomy is feasible with either cisplatin or mitoxantrone. However, many of these patients still recurred and prospective trials will be required to determine which (if any) consolidation intraperitoneal treatment is best following a negative secondlook laparotomy.

Primary carcinoma in situ of the vagina

Primary carcinoma in situ of the vagina is very rare in Japan. The patient is 57 years old and its found in screening of cervical cancer. Carcinoma in situ was located in the upper and posterior wall of the vagina. No cancer was seen in the cervix and vulva. The patient had extenden hysterectomy. It is necessary to emphasize the need for inspection of the vaginal wall and vaginal smear at the screening of cervical cancer.