Nagayasu Toyoda

Screening tests for gestational diabetes in Japan in the 1st and 2nd trimester of pregnancy

The objective of this study was to investigate the utility and characteristics of various screening procedures for gestational diabetes mellitus (GDM) in Japan during the first trimester and between 24 and 28 weeks of pregnancy. The subjects were 749 pregnant women who came to our hospitals. A 50-g oral glucose challenge test (GCT), casual plasma glucose measurements, fasting blood glucose measurements, and glycosylated hemoglobin measurements were performed in the first trimester. Subjects with no abnormalities were tested again at 24-28 weeks of gestation. Of the 749 subjects, 22 (2.9%) tested positive for GDM. Of those 22 patients, 14 were diagnosed with GDM in the first trimester (63.6%) and eight in the second trimester (36.4%). This finding suggests the importance of screening for glucose intolerance in the first trimester. Furthermore, it appears that the GCT has the most utility for GDM screening; the other screening methods tested were not as useful because of their low sensitivity, particularly in the second trimester.

A novel human HER2‐derived peptide homologous to the mouse Kd‐restricted tumor rejection antigen can induce HLA‐A24‐restricted cytotoxic T lymphocytes in ovarian cancer patients and healthy individuals

A mouse HER2‐derived peptide, HER2p63 (A) (TYLPANASL), can induce Kd‐restricted mouse cytotoxic T lymphocytes (CTL) and also function as a tumor rejection antigen in an in vivo assay. Since the anchor motif of mouse Kd for peptide binding has much similarity to that of human HLA‐A2402, we asked if human HER2p63 (T) (TYLPTNASL) could induce HER2‐specific CTL in HLA‐A2402‐positive individuals. Peripheral blood mononuclear cells (PBMC) of HLA‐A2402‐positive individuals were sensitized in vitro with HER2p63‐pulsed autologous dendritic cells prepared from PBMC. CTL clone derived from these specifically lysed HER2‐expressing cell lines bearing HLA‐A2402. Cytotoxic activity of the CTL clone against the HER2‐expressing cell line bearing HLA‐A2402 was blocked by antibodies against CD3, CD8, HLA‐A24 or MHC class I, and was also inhibited by the addition of excess HER2p63‐pulsed C1R bearing HLA‐A2402. Killer cells were generated from PBMC of seven healthy individuals and five ovarian cancer patients, all of HLA‐A2402 type, by in vitro sensitization with HER2p63‐pulsed autologous antigen presenting cells. These killer cells selectively lysed HER2‐expressing SKOV3 transfected with HLA‐A2402 cDNA, indicating high immunogenicity of HER2p63 in all 12 individuals examined.

The ex utero intrapartum treatment (EXIT) procedure in giant fetal neck masses.

Giant fetal neck masses can cause airway obstructions with potential poor fetal prognosis after delivery. The relationship between the fetal neck mass and airway structure can be defined prenatally with ultrasound and magnetic resonance imaging (MRI). The ex utero intrapartum treatment (EXIT) procedure is an available technique to obtain a fetal airway while feto-maternal circulation is preserved. We present a case in which prenatally a giant fetal neck mass was diagnosed on ultrasound and MRI, and a successful EXIT procedure could be performed.

Expression of glucose transporter 4 mrna in adipose tissue- and skeletal muscle of ovariectomized rats treated with sex steroid hormones

The effects of 17beta-estradiol (E) and/or progesterone (P) on glucose transporter 4 (GLUT4) expression in the adipose tissue and skeletal muscle of ovariectomized female rats were studied. The Sprague-Dawley rats received daily subcutaneous injections of various doses of E and/or P for 7 days (n=5-6 per dose). The expression of GLUT4 mRNA was assessed by performing ribonuclease protection assays. GLUT4 protein levels were assessed by Western blotting assays. The adipose tissue levels of GLUT4 mRNA were reduced by the administration of 50 microg E, which resulted in unphysiologically high serum E concentrations. Although the GLUT4 mRNA levels did not change after the administration of 10 microg E or 5 mg P, they were reduced significantly to approximately half the control group level by the administration of both hormones (p <0.01). The skeletal muscle GLUT4 mRNA levels were not changed significantly by hormone treatment. These findings suggest that E and P may be involved in the regulation of GLUT4 mRNA expression in adipose tissue.

Expression of Tenascin-C in Stromal Cells of the Murine Uterus During Early Pregnancy: Induction by Interleukin-1α, Prostaglandin E2, and Prostaglandin F2α

Abstract Tenascin-C (TN-C), an extracellular matrix glycoprotein, is known to be expressed in uterine stroma in the peri-implantation period. Examination of the spatiotemporal pattern during early pregnancy using immunohistochemistry and in situ hybridization revealed TN-C expression in the stroma beneath the luminal epithelia of the murine endometrium on Days 0 and 1 of pregnancy, subsequent disappearance, and reappearance on Day 4. After decidualization, tissue around the deciduoma was positive. In situ hybridization demonstrated TN-C production by the stromal cells adjacent to the epithelia. To investigate the regulation of TN-C expression in vitro, murine uterine stromal and epithelial cells were isolated and cultured. Addition of interleukin-1α (IL-1α) and prostaglandin E2 (PGE2) and F2α (PGF2α) induced TN-C expression in the stromal cells at both protein and mRNA levels, while the sex steroid hormones, progesterone and β-estradiol, exerted little effect. Immunohistochemistry using anti-IL-1α antibody showed epithelial cells to be positive on Days 2–4 of pregnancy, and addition of progesterone but not β-estradiol enhanced IL-1α expression in epithelial cells in vitro. In a culture insert system, TN-C expression by stromal cells cocultured with epithelial cells was induced by addition of progesterone alone that was blocked by additions of anti-IL-1α antibody. Collectively, these findings indicate that TN-C expression in the preimplantation period is under the control of progesterone, but not directly, possibly by the paracrine and autocrine intervention of IL-1α secreted by epithelial cells and PGE2 and PGF2α secreted by stromal cells.

Aspiration of Giant Hepatic Cyst in the Fetus in utero

A large hepatic cyst was diagnosed in a fetus at 29 weeks of gestation by puncture and aspiration performed under ultrasonographic guidance. This procedure was repeated five times at about 2-week intervals, with a total of 1,446 ml of cystic fluid aspirated. A healthy female infant was delivered vaginally at 40 weeks of gestation. Her Apgar score was 9 at 1 min, and she weighed 2,790 g. On the 14th postnatal day, the hepatic cyst was punctured percutaneously under ultrasonographic guidance, and 119 ml of yellow fluid was aspirated. The cyst was subsequently not visible on computed tomography at 9 months. Surgery was not indicated in this infant, and the cyst was also not detected at the age of 21 month. Thus, treatment in utero appeared to have obviated the need for surgery.

Pregnancy and Delivery in Patients with von Willebrand's Disease

Objective: To explore the obstetrical problems that are encountered in women with von Willebrands disease (vWD).

Comparison of the validity of the criteria for gestational diabetes mellitus by WHO and by the Japan Society of Obstetrics and Gynecology by the outcomes of pregnancy

Data for 416 Japanese pregnant women who received a 75g oral glucose tolerance test (OGTT) for determination of gestational diabetes mellitus (GDM) in 13 hospitals in Japan were analyzed retrospectively. Comparison of the diagnostic criteria of the World Health Organization (WHO) and the Japan Society of Obstetrics and Gynecology (JSOG) revealed pregnant women who met the latter criteria for GDM to have significantly higher incidences of low Apgar scores, respiratory problems, neonatal hypoglycemia, preterm delivery and requirements for insulin therapy and cesarean section. The women who met the WHO criteria but not the JSOG criteria had minor complications. These observations suggest that the GDM criteria of the JSOG are more appropriate than the WHO criteria from the standpoint of therapeutic intervention for pregnant women.

Possible direct effect of gonadotropin releasing hormone on human endometrium and decidua.

Decidualization of endometrial tissues, which is essential for implantation and the continuation of pregnancy, is induced by pituitary hormones that are regulated by gonadotropin releasing hormone (GnRH). Our objective was to determine the role of a direct action of GnRH on endometrial tissues by comparing the characteristics of receptors for GnRH in human endometrial and decidual tissues. Competitive binding studies were performed with the protease-resistant GnRH analogues, buserelin and [125I] buserelin. The effects of buserelin on phosphoinositol turnover were determined by the measurement of inositol 1,4,5-triphosphate(IP3). The values for the dissociation constant (Kd) and number of binding sites (Bmax) per unit protein versus buserelin for endometrial tissues did not differ from the values for decidual tissues. However, the Bmax per unit DNA was significantly higher in endometrial tissues. Also, buserelin induced a significant increase in IP3 in decidual tissue. These results indicate that GnRH may be a potential modulator of the function in human endometrium and decidua. The signal transduction mechanism for GnRH action appeared to involve the accelerated turnover of phosphoinositol.

Mutational Analysis of BRCA1 Gene in Ovarian and Breast-ovarian Cancer Families in Japan

We analyzed the alteration of BRCAI in DNA obtained from 83 individuals of 13 Japanese site‐specific ovarian cancer families and 6 breast‐ovarian cancer families. Six germline mutations were detected in 7 families, which consisted of 4 breast‐ovarian cancer and 3 site‐specific ovarian cancer families, by single‐strand conformation polymorphism analysis, followed by direct sequence determination. The mutations included three framcshifts, two nonsense mutations, and one missense mutation causing loss of a zinc‐binding motif. The frequency of loss of heterozygosity at the microsatellite markers on the BRCAI gene was 57% (8 of 14 cases) in site‐specific ovarian cancer families, and 100% (6 of 6 cases) in breast‐ovarian cancer families. All tumors of the patients carrying a mutation of BRCAI showed deletion of wild‐type alleles, implicating BRCAI as a tumor suppressor gene. Tbese results suggest tbat germline mutations of the BRCAI gene play an important role in the carcinogen‐esis of breast and/or ovarian cancer in a majority of breast‐ovarian cancer families and in some site‐specific ovarian cancer families.