[Ultrastructural observations of follicular oocytes inseminated in vitro in squirrel monkeys (Saimiri sciureus)].

The ultrastructure of squirrel monkey (Saimiri sciureus) follicular oocytes inseminated in vitro was studied at the Endocrine Research Center of Michigan State University in the United States. Adult female squirrel monkeys were induced to ovulate with FSH-hCG, followed by laparoscopy 16 hours after hCG to collect ovarian follicular oocytes. A total of 45 oocytes were inseminated 21 hours after recovery by adding sperm collected by electroejaculation, followed by insemination culture for 24 hours. Inseminated oocytes were examined by light microscopy and transmission electron microscopy. Only three (6.7%) of inseminated oocytes were judged to be fertilized morphologically, but no sperm remnant was found in the cytoplasm of the oocytes. Fifteen (33.3%) had reached the second metaphase stage. As a whole, the maturation rate was 40%. Ultrastructurally, acrosome reacted sperm were observed in the cumulus cell layer, confirming the occurrence of sperm capacitation in the in vitro culture system. Cortical granules were seen throughout the cytoplasm of the germinal vesicle oocyte, especially in the vicinity of the Golgi complexes, suggesting the possibility that cortical granules originate from Golgi complexes. Metaphase chromosome, first polar body and cortical granules were observed at the second metaphase oocyte. Of three oocytes judged to be fertilized, one oocyte contained six pronucleus-like structures and another oocyte contained a pronucleus supposing fusion of two pronuclei (syngamy). But the ultrastructural morphology of a normal appearing fertilized oocyte could not be obtained. The possible cause of the low fertilization rate in the present study is supposed to be the poor follicular response and the prematurity of the cytoplasm due to repeated administrations of exogenous gonadotropins.

Apoenzyme of aspartate aminotransferase isoenzymes in maternal plasma during labor and following delivery.

In eight normal mothers the effects of pyridoxal 5'-phosphate (PLP) addition on the plasma activities of aspartate aminotransferase isoenzymes (AST-s and AST-m) during labor and following delivery were investigated. The AST-s activities with and without reactivation by PLP appeared to increase immediately after delivery and they were even higher on the 4th day postpartum. On the other hand, there were significant elevations in both the AST-m activities immediately and at 2 h after delivery, but not on the 4th day postpartum. Of AST isoenzymes measured in the nonstress test, only the relative activation rate of AST-m by PLP added was significantly higher than the control value. The present study may come to the following conclusions: 1) The relative activation rate of plasma AST-m activity by PLP may be a reliable index of vitamin B6 nutritional status during pregnancy. 2) The increases in AST-m activity with and without PLP added during labor suggest a minimal damage of mitochondria in skeletal, cardiac and uterine muscle cells. 3) The AST-s isoenzyme determinations with and without PLP may be especially useful as sensitive indication of erythrocyte and/or liver damage after delivery.

[Clinical studies on cefuzonam in the field of obstetrics and gynecology].

Cefuzonam (CZON, L-105), a cephem type antibiotic, was clinically studied in the field of obstetrics and gynecology. The results are summarized as follows: CZON 1-2 g was administered by injection twice daily to 11 cases of infections (4 of endometritis, 2 of pyometra, 3 of adnexitis and peritonitis, 1 of abdominal abscess, and 1 of puerperal fever). Clinical efficacy was excellent in 2 cases and good in 9 cases, with a very high overall efficacy rate of 100%. Slight elevations of GOT and GPT in 1 case were noted in clinical laboratory tests. No side effects attributable to the drug were noted. CZON is considered to be a useful drug for obstetric and gynecological infections.

Maternal blood distribution of zinc and copper during labor and after delivery.

In 15 normal mothers the stress effects of labor on zinc and copper distributions in erythrocytes and plasma were investigated. Lower levels of total zinc and of zinc derived from both carbonic anhydrase isoenzymes (CA-I and CA-II), especially from CA-I, in erythrocytes were obtained immediately after delivery. On the other hand, there was a significant increase in total plasma zinc concentration only on admission to hospital in labor, although albumin-bound zinc concentration remained elevated throughout the entire labor. These results suggest that there are unexplained disparities between the changes in zinc levels in maternal erythrocytes and plasma during labor.

Maternal plasma concentrations of catecholamines and cyclic nucleotides during labor and following delivery.

In plasma obtained from seven mothers before, during, and after normal labor and delivery, catecholamine and cyclic nucleotide concentrations were investigated. Dopamine concentration showed a significant elevation on admission to hospital in labor and there were marked increases in norepinephrine and epinephrine concentrations during labor at 10 cm cervical dilatation and immediately after delivery, respectively. No significant change in dopa, cAMP, or cGMP was found during the experimental period. However, since the van Beaumont quotient (J.Appl. Physiol. 34, 102-106) for cAMP did not follow the reduction in plasma volume, the concentration appeared to rise during labor. Positive correlations were observed between epinephrine on one hand, and heart rate and systolic blood pressure on the other, as well as between norepinephrine and cAMP, respectively, during labor. The diminution of epinephrine on the fourth day postpartum might reflect a reduction of emotional stress concomitant with labor. cAMP was found to be quickly cleared from the bloodstream within 2 hours after delivery. These results suggest that plasma concentrations of epinephrine and cAMP, especially epinephrine, are indices of maternal psychological and physiological stress during labor and following delivery.

[Fundamental and clinical studies on aztreonam in obstetrics and gynecology].

Aztreonam (AZT), a monobactam antibiotic, is known to have a high activity against Gram-negative bacteria. Fundamental and clinical studies were carried out on AZT with the following results. Following 1 g of bolus intravenous injection, the transfer of AZT to uterine artery and internal genital organs was found to be satisfactory. The levels of the drug in uterine artery showed 34.28, 4.50 micrograms/ml at approximately 2, 6 hours, and those in internal genital organs showed 3-34 micrograms/g at 2 hours. Clinical efficacy was; excellent in 4 cases, good in 13 cases and poor in 1 case, with the very high overall efficacy rate of 94.4%. Abnormal laboratory findings and side effects due to the drug were not noted.

[Chemotherapy of ovarian cancer based on in vitro (HTCA) and in vivo (nude mice) chemosensitivity testing].

The human tumor clonogenic assay (HTCA) and the human tumor xenograft system implanted in nude mice were performed simultaneously in an ovarian cancer patient as chemosensitivity testing. Eight anticancer drugs (5-FU, MMC, VCR, ACD, BLM, VLB, CDDP, and ADM) were applied to the HTCA and the human tumor xenograft system. In the HTCA, 5-FU and MMC were sensitive, VCR was moderately sensitive, and ACD, BLM, VLB, CDDP, and ADM were resistant. In the human tumor xenograft system, MMC, VCR, and ADM showed tumor regression (++), and CDDP, VLB, BLM, 5-FU, and ACD exhibited no response (-). Two of the three drugs, which were classified as sensitive or intermediately sensitive in the HTCA, showed tumor regression (++) in the human tumor xenograft system. And four of the five drugs, which were resistant in the HTCA, exhibited no response (-) in the human tumor xenograft system. Clinically, PVB therapy (CDDP, VLB, and BLM) was applied to the present patient, but after recurrence, 5-FU + MMC therapy was applied on the basis of the results of the HTCA. In addition, ADM was added with reference to the results of the human tumor xenograft system. As a result of this therapy, the tumor growth was inhibited. It is possible from the present data that simultaneous chemosensitivity testing of the HTCA and the human tumor xenograft system implanted in nude mice is very useful when choosing sensitive anticancer drugs.

[Experimental and clinical studies of cefmenoxime in the field of obstetrics and gynecology].

The study group was organized to evaluate the usefulness of cefmenoxime (CMX) injection, a new synthetic cephalosporin, for the treatment of infections in the field of obstetrics and gynecology. Fundamental and clinical studies were made by the society and the following results were obtained. 1. The peak distribution of CMX's MIC for E. coli, Klebsiella sp., Enterobacter sp., Bacteroides sp. and Peptococcus sp. isolated from obstetrical and gynecological infections with relatively high frequencies area 0.1, less than or equal to 0.05, 0.2, 3.13, 1.56 micrograms/ml, respectively, with an inoculation of 10(6) cells/ml. 2. When 1 g of CMX is administered by intravenous drip infusion for 1 hour, the maximum concentrations in various tissues of female genital organs were as follows: 14.2 and 13.2 micrograms/g in ovary and oviduct, respectively, at 1.20 hours after the start of administration, and 16.9 and 26.3 micrograms/g in corpus uteri and cervix uteri, respectively, after 1 hour. As for the transfer to the exudate in the pelvic dead cavity, the peak concentration was 15.6 micrograms/ml after 2.13 hours. 3. In the clinical studies, CMX was given to 258 cases with female genital organ infections and others. As for the clinical effects, with exclusion of 3 cases in which other antibiotics are concomitantly used, responses were excellent in 76 cases, good in 162 cases and poor in 17 cases, among 255 cases in total. The efficacy rate was 93.3%. The efficacy rates by diseases were 97.1% (68/70) for intrauterine infections, 88.8% (79/89) for intrapelvic infections, 98.4% (62/63) for adnexitis, and 100% (23/23) for infections of external genital organs. As for the clinical effects on causative bacteria, the efficacy rates were 100% (19/19) for single infections due to Gram-positive bacteria, 94.8% (55/58) for single infections due to Gram-negative bacteria, and 88.2% (15/17) for single infections due to anaerobic bacteria. And its efficacy rates were 89.6% (69/77) for mixed infection cases. Side effects were observed in 2 cases (0.8%); 1 case with eruption, and 1 case with diarrhea and vomiting. As for abnormal laboratory findings, lower white blood cell count was observed in 2 cases and elevation of the values regarding hepatic functions in 9 cases. All cases were returned to the normal after the completion of the administration. Cefmenoxime showed a satisfactory clinical efficacy and a potent bacteriological effect in treatment of the infections in the field of obstetrics and gynecology, and it has been concluded that cefmenoxime will be useful addition to the antibiotics for the therapy of these infections.

A sensitive radioreceptor assay for follicle stimulating hormone with PMS-primed immature rat ovary.

After a single PMS (50 IU) injection to 25-day-old rats, FSH receptor content of the ovarian tissue increased progressively for 4 days, then showed a tendency to decrease, while LH receptor content remained unchanged for 3 days, then gradually increased. From these facts, we established a radioreceptor assay system, employing 3,000 rpm precipitates of homogenates of the ovaries obtained 3 days after PMS injection as the receptor preparation. The dissociahe standard curve was obtained with 0.125--16 ng/tube of NIAMDD rat FSH I-3. Purified preparations, NIAMDD rat LH I-4 and NIAMDD rat TSH I-4 influenced the binding only at high concentrations possibly owing to FSH contamination. When the anterior pituitary homogenates obtained from rats in the various physiological states were assayed by this system, the intra-assay coefficient of variation and inter-assay coefficient of variation were 7.5% and 13.7%, respectively, and the assay values were well correlated with those obtained by radioimmunoassay (r = 0.988, the slope of the regression line = 1.14).