Kohki Yamashita

A study of zinc distribution in erythrocytes of normal humans

SummaryThe assignment of zinc bound to carbonic anhydrase isoenzymes (CA-I and CA-II) and Cu2 Zn2 superoxide dismutase (SOD1) was investigated in the hemolysates from 21 normal male subjects.Sufficient care was taken to remove leukocytes and platelets. The following values of zinc distribution were obtained:total zinc, 1113.8±22.7 (mean±S.E.) μg · 100 ml−1; CA-I-derived zinc, 866.6±26.2 μg · 100 ml−1; CA-II-derived zinc, 99.9±3.9 μg · 100 m−1; SOD1-derived zinc, 60.3±1.9 μg · 100 ml−1; the other zinc, 87.0±12.6 μg · 100 ml−1. Namely, 7.6% of the zinc in human erythrocytes is not bound to the carbonic anhydrases and Cu2Zn2 superoxide dismutase, but present in available form or attached to other enzymes.

Possible role of prostaglandin F in blastocyst implantation

The synthesis and release of Prostaglandin F (PGF) by the rabbit blastocyst and endometrium were investigated on Day 6 and Day 7, using radioimmunoassay, autoradiography and conversion experiments. The following results were obtained: The content of PGF in the blastocyst increased significantly (P less than 0.01) from Day 6 to Day 7. The content of PGF in the endometrium was significantly higher (P less than 0.05) on Day 7 implantation sites compared to the other areas. The in vitro synthesis and release of PGF by Day 6 blastocysts sharply increased after one and two hours of culture, respectively. Thereafter both values declined with time. The in vitro synthesis and release of PGF by Day 6 endometria increased continuously with time. 14C-arachidonic acid (14C-AA) was incorporated into Day 6 blastocysts in vitro and converted to PGF2 alpha. These results suggest that both the endometrium and the blastocyst are the sources of the PGs involved in implantation, and that PGF derived from the blastocysts may act as the trigger of implantation.

Serum immunosuppressive substance in patients with gynecologic malignancies and in pregnant women.

The presence of serum immunosuppressive substance (IS) was determined in 134 patients with malignant gynecologic tumors (105 cervical cancers, 15 endometrial cancers, and 14 ovarian cancers), 45 patients with benign gynecologic tumors (33 uterine myomas and 12 ovarian tumors), 10 patients with severe inflammatory diseases, 326 pregnant women, and 48 healthy controls. The mean levels and percentages of positive levels (>750 μg/ml) in both the groups of cancer patients and the group of patients with severe inflammatory diseases were significantly higher than those in the groups of patients with benign tumors or the control group. In pregnant women, however, the majority of serum levels were within a normal range, showing relatively elevated levels in the first trimester. In the patients with malignancies (cervical, endometrial, and ovarian cancers), extremely elevated levels (>1000 μg/ml) suggested an active or a progressive state of the malignancies. It became clear that serial IS determinations are valuable for monitoring the disease state or judging the effect of therapy.

ras oncogene product p21 expression and prognosis of human ovarian tumors

Monoclonal antibody rp-28 directed against the ras gene product p21 has been studied to evaluate ras p21 expression in malignant and benign ovarian tissues. Some ovarian carcinomas of serous and mucinous cystadenocarcinomas, undifferentiated adenocarcinomas, and clear cell carcinomas demonstrated intense staining of ras p21. The frequency and intensity of ras p21 staining were observed to increase with the degree of malignancy. There was no significant difference in ras p21 expression between early and late stages in ovarian tumors arising from the coelomic epithelium. With respect to prognosis, no differences between the ras p21-positive and -negative cases in ovarian tumors arising from the coelomic epithelium were observed. It is, therefore, possible to say that ras p21 expression was not related to clinical staging and prognosis. Expression of ras p21 in malignant lesions was higher than that in benign lesions of the ovary, and the expression is associated with the degree of malignancy in some types of ovarian tumors. Overexpression of ras p21 was observed in epithelial tumors; however, increased expression was not observed in germ cell and sex-cord stromal tumors. This differential expression of ras p21 is due to the different histogenesis of ovarian tumors. This fact may reflect a different carcinogenic mechanism for different types of malignancy.

HLA antigens in husband-wife pairs with trophoblastic tumor☆

Abstract Seventy-four patients with trophoblastic tumors (49:hydatidiform moles, 16:invasive moles, and 9:choriocarcinomas), 68 of their husbands (45, 15, and 8 respectively), and 76 controls were typed for HLA A and B specificities to determine whether or not there are specific HLA antigens associated with the occurrence of this tumor, and whether there is a relationship between the histocompatible combination of patient-husband and tumor growth. All subjects were Japanese. The results obtained were as follows. (a) No paternal and maternal HLA antigen was demonstrated to be associated with the tumor. (b) Histocompatibility between patient and husband was not responsible for tumor growth.

Absence of major histocompatibility complex antigens in choriocarcinoma.

Human trophoblastic cells are of fetal origin, and are not rejected or recognized as a target by cytotoxic T cells. The mechanism for this may not be easily explained, but whether trophoblastic cells express major histocompatibility complex antigens remains a most important topic. The recent introduction of monoclonal antibodies of defined major histocompatibility complex specificities has clarified that two types of trophoblastic cells are present in normal human early placenta.’ The one is a villous trophoblast (syncytiotrophoblast) which lacks both class I and class II major histocompatibility complex antigens, and the other is a extravillous trophoblast (nonvillous cytotrophoblast) which expresses at least class I major histocompatibility complex antigens. These findings stimulated us to analyze choriocarcinoma since it is a ma1ignar.t tumor

Studies of Enzymuria and β2-Microglobulinuria in Patients Receiving Prophylactic Cefotaxime in Obstetric and Gynaecological Surgery

SummaryIncreased urinary excretion of β2-microglobulin and certain enzymes, for example leucine aminopeptidase (LAP) and N-acetyl-β-D-glucosaminidase (NAG), produces sensitive markers of damage to renal tubular cells. These markers were monitored in patients undergoing obstetric and gynaecological surgery who received cefotaxime for 5 days. Slight increased activity in NAG and β2-microglobulin was observed in some patients 3 to 5 days after surgery. However, no changes of significance were found in levels of urinary LAP and NAG. During cefotaxime treatment, urinary β2-microglobulin in the group of abdominal hysterectomy increased slightly. In most cases renal side effects are negligible and cannot be detected by these tests. These results indicate that administration of cefotaxime in obstetric and gynaecological surgery does not produce cumulative renal tubular toxicity.

Anti-HLA antibodies detected in the serum of patients with gestational trophoblastic tumors (choriocarcinomas and invasive moles)

Abstract Four patients with Choriocarcinomas and nineteen patients with invasive moles were analyzed seriately for anti-HLA antibodies. One serum out of four with Choriocarcinomas and 6 sera out of 19 with invasive moles demonstrated monospecific anti-HLA antibodies and persisted as long as each patient was treated by chemotherapy. One serum which was not determined to be anti-HLA antibody was checked with the serum of one patient with choriocarcinoma. The relationship between the antibodies and tumor growth or response to chemotherapy was studied clinically, but no definite relationship was established. In the present studies, antigenicity of the trophoblastic cell itself was not established.

Seasonal variations in the responses of glycolytic intermediates of human erythrocytes to acute cold exposure

Seven male students were studied to observe the effects of acute cold exposure (at 10°C for 60 min) on erythrocyte concentrations of glycolytic intermediates in summer and in winter. The subjects shivered slightly but frankly in both experiments. Significant decreases were observed in the concentrations of pyruvate and lactate during body cooling in summer, but not in winter. The lactate concentration remained significantly reduced 15 min after cold exposure. After 60 min of cold exposure in summer, a negative crossover point appeared to exist between phosphoenolpyruvate and pyruvate and erythrocyte pyruvate kinase activity showed a significant decrease. No seasonal difference was observed in the initial control values of the intermediates measured. From these results and the fact that glucose, pyruvate and lactate are evenly distributed between erythrocytes and plasma, it is likely that erythrocytes and skeletal muscles need less fuel substrate, glucose during cold exposure in winter than in summer, suggesting that an increased economy of energy for homeostasis is achieved.

Ultrasonic and endocrinologic investigation of ovarian follicle and free fluid in cul-de-sac for monitoring follicular development and ovulation

The accuracy of ultrasound as a means of monitoring follicular development and ovulation detection was studied in 31 infertile women. The follicular diameter measured by ultrasound correlated well actual diameter found during laparotomy (r = 0.88), and estradiol‐17β (E2) and progesterone in follicular fluid aspirated from large follicles (≧ 16 mm) indicated high levels. Furthermore, in 23 out of the 55 cycle, ultrasound examinations demonstrated free fluid in cul‐de‐sac at ovulation, and concentrations of E2 and progesterone (P) in peritoneal fluid by the puncture of cul‐de‐sac dramatically increased after ovulation. We suggested that ultrasound monitoring provided a reliable measure of follicular growth and prediction of ovulation, and a combined use of ultrasound and puncture of cul‐de‐sac could be developed into a new tool for the study of the mechanism of human ovulation.