We report the case of a 62-year-old woman who was admitted to our hospital with diabetic ketoacidosis. Her urinary C-peptide was 3.5μg/day, HLA typing was DR9, and serum was positive for islet cell antibodies. There was no significant increase in the major viral titer. Pancreatic head tumor was suspected, and pancreaticoduodenectomy was performed. The pathology of this tumor was polycystic adenoma. We examined the surgical specimen from around the tumor histologically. The pancreatic islets had decreased in number. The immunohistochemical staining of islets for insulin, glucagon and somatostatin showed that the number of B cells had decreased remarkably, while A and D cells were preserved Marked lymphocytic infiltration was observed in the islets. The majority of lymphocytes were helper/inducer and suppressor/cytotoxic T cells, which did not express HLA-DR antigen or interleukin-2 receptor. No NK cells were present in the islets. The present case, which was examined histologically in detail, is consistent with the previously proposed hypothesis that autoimmunity might play an important role in the pathogenesis of insulin-dependent diabetes mellitus.

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Epidermal growth factor (EGF) Immunoreactivity, EGF binding activity, and their ontogenic changes were investigated in placenta, amnion, and various organs of mouse fetus and neonate. EGF immunoreactivity was detected in all organs examined such as fetal and neonatal liver, brain, lung, intestine, placenta, and amnion. The fetal tissues having relatively higher EGF content were mnion, liver, and intestine. The tissue distribution of EGF immunoreactivity in the fetus did not differ from that in neonate. As for its ontogenesis, EGF immunoreactivity was almost constant during fetal development in liver, brain, and placenta, whereas it gradually increased in intestine. Scatchard plot analysis of EGF receptor, which was also demonstrated in all organs studied, revealed that EGF receptor in these tissues

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Since 1968, a total of 23 patients with multiple endocrine neoplasia type 2B (MEN 2B) have been identified in Japan. The mean age at diagnosis was 20.3 years (range, 6 to 39 years). All patients had neuromas and bumpy lips. All patients underwent thyroidectomy for medullary thyroid carcinoma (MTC). Ten of 23 patients had pheochromocytomas. One patient died of cerebral bleeding at the age of 43, 2 patients died of MTC at the age of 35 and 12. Five-, 10-, and 15-year survival rates were 100%, 92%, and 88%, respectively. The clinical course of MTC in MEN 2B in Japanese is not so aggressive when compared with about a 50% 10-year survival reported for Caucasians. Genetic nalysis of 4 MTC and 1 pheochromocytoma from 4 patients with MEN 2B revealed no common changes in regard to loss of heterozygosity on 21 chromosomes or point mutations of the ras, Gsα, or p53 genes.

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To investigate the changes in the serum androgen concentrations and the Free Androgen Index (FAT) in women during danazol therapy, we measured the serum concentrations of adrenal steroids and danazol metabolites, and then examined the effects of danazol metabolites on assays for serum androgens. Thirteen women who had endometriosis were treated with danazol (300 or 400 mg/day) for 8 to 16 weeks. Blood samples were taken before, during, and after the medication. During the danazol therapy, serum testosterone (T), cortisol (F), and sex-hormone binding globulin (SHBG) significantly decreased (P<0.05); but serum dehydroepiandrosterone-sulfate (DHEAS) and FAI increased (P<0.05). The serum concentrations of danazol metabolites were: danazol, 209.0±28.3 (ng/mL, mean±SEM); ¹-2-hydroxymethyl ethisterone, 114.4±8.4; and 2-hydroxymethyl ethisterone, 660.0±54.2. There was considerable cross-reaction between danazol metabolites and androgens [T, androstenedione (A), and dehydroepiandrosterone (DHEA)] in the direct assays. As for the ratios of adrenal steroids in serum, the DHEAS/F, DHEAS/DHEA, and 11-deoxycortisol (S)/F ratios increased (P<0.05). We conclude that the increase in FAT and DHEAS represents increased native androgenic activity with danazol, and the changes in adrenal steroid ratios in serum indicate the inhibition of 11β-hydroxylase and sulfatase activities during danazol therapy.

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Superovulation with exogenous gonadotropins has been used widely to study the reproductive events in animals and in vitro fertilization and embryo transfer in human beings. However, details of the mechanism of superovulation are not yet clearly understood. The present study was conducted to study the mechanism of superovulation with purified human FSH and LH preparations. Cyclic hamsters received continuous infusion of purified FSH and LH for 4 days, from estrus to proestrus. The number of ova shed was significantly increased by 5 iu FSH/minipump infusion, and further increased by 5iu FSH/minipump plus 5 iu LH/minipump group. On the other hand, the number of atretic follicles was decreased significantly (P<0.01) by the combined infusion of FSH and LH, but not by FSH alone. Among these experimental groups, there was no significant difference in embryonic development which was assessed by the counting of the number of 4-cells and more developed embryos on day 3 of pregnancy. These data indicated the different roles of FSH and LH in the superovulation process, because FSH stimulated some small follicles to grow and LH, in the presence of FSH, rescued the follicles to undergo atresia.

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20α-Hydroxysteroid dehydrogenase (20α-HSD) (EC.1.1.1.149) is the enzyme which catabolizes progesterone to 20α-dihydroprogesterone (20α-OHP), a biologically inactive steroid, and is distributed in a variety of tissues including non-steroid-producing tissues. In the present study, changes in cytosolic 20α-HSD activity were investigated in rat placental tissues and its relationship to embryonic mortality was considered; the mesometrial endometrium (including the ectoplacental cone) on days 8-11 of pregnancy (day 0=estrus), and the chorioallantoic placenta and visceral yolk sac (vitelline membrane) on days 12-21 were separately subjected to measurement of the enzyme activity. 20α-HSD activity was not detected in the chorioallantoic placenta until day 20 and then increased dramatically on day 21. Interestingly, considerable activity of the enzyme was found in the visceral yolk sac from days 14 to 21 and in the mesometrial endometrium from days 8 to 10, whereas it was undetectable in these tissues on days 11 and 12. Analysis of DEAE column chromatography revealed that these tissues contain two different types of 20α-HSD (HSD-1 and HSD-2). By an immunohistochemical method, with polyclonal antiserum to rat 20α-HSD, decidual cells and trophoblastic giant cells adjacent to the ectoplacental cone (day 10), spongiotrophoblasts and visceral yolk sac cells (days 21) were positively stained. The number of fetuses on day 10 of pregnancy was 15.4 and decreased significantly to 12.9 on day 12. Since the progesterone concentration in the amniotic fluid was only 3-4% of the maternal serum concentration, expression of 20α-HSD in the placental tissues seems to lower the concentration of progesterone in the fetal environment. Thus, 20α-HSD activity was found in the tissues surrounding the fetus during pregnancy in the rat except for the specific period (days 11 and 12) during which spontaneous fetal loss occurs. 20α-HSD is therefore present in the decidual cells during placentation and in the visceral yolk sac throughout pregnancy. These results support the hypothesis that 20α-HSD acts as a survival factor for fetuses, protecting them from the cytotoxic activity of progesterone by metabolizing it to the inactive steroid.

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Thyroid peroxidase (TPO) in 4 benign tumors, 9 malignant tumors and 7 Graves' goiters was examined both by biochemical methods (guaiacol and iodide oxidation assays) and histochemical methods (indirect immunofluorescence methods with polyclonal anti-TPO antibodies). The protein- based specific activity of TPO of Graves' goiters was comparable to or higher than that of normal thyroid tissues, while that of benign and malignant tumors were comparatively lower and variable. TPO was stained with the polyclonal anti-TPO antibodies in the cytoplasm of all benign adenomas, Graves' goiters and follicular carcinomas, but not of papillary carcinoma and metastatic cells. TPO staining markedly appeared in apical areas of Graves' goiters but was scarcely detected in the areas of other disordered thyroids. Polyclonal anti-thyroglobulin antibodies detected thyroglobulin in the follicular lumen of these diseased thyroids except in metastatic carcinoma cells.

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In the present study, we examined the effect of basic fibroblast growth factor (bFGF) on gonadotropin-dependent estradiol and progesterone production in cultured rat immature and FSH-primed granulosa cells. Treatment with bFGF alone did not affect the biosynthesis of estradiol and progesterone in immature and FSH-primed granulosa cells. Basic FGF significantly inhibited FSH-induced estradiol and progesterone production in both immature and FSH-primed granulosa cells. In contrast, bFGF exerted a significant stimulatory effect on LH-induced estradiol and progesterone production in FSH-primed granulosa cells. Our finding that bFGF exerts opposite effects on gonadotropin-dependent steroidogenesis suggests that bFGF may play important paracrine and/or autocrine roles in the process of follicular development, ovulation, and subsequent luteinization.

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The effects of calyculin-A (CL-A), a phosphatase inhibitor, on 20α-hydroxysteroid dehydrogenase (20α-HSD) activity were studied in rat luteal cell cultures to examine whether protein phosphatases were involved in the regulation of the enzyme activity. Luteal cells were harvested from the rats on day 6 of pseudopregnancy. In the absence of prolactin (PRL), the stimulatory effect of CL-A (10^-9M) on 20α-HSD activity was observed during the first 3h period of the culture, but was not discernible thereafter. Since the enzyme activity of luteal cells during pseudopregnancy is known to be suppressed by PRL, the results suggest that this effect of CL-A could be manifested only as long as the effect of endogenous PRL was remained. On the other hand, CL-A enhanced the 20α-HSD activity of the cells cultured for 24h, where a spontaneous increase in 20α-HSD activity was blocked by exogenously supplementing the culture medium with PRL. These results suggest that a CL-A sensitive phosphatase(s) may be involved in the action of PRL in suppressing the increase in 20α-HSD activity in rat luteal cells.

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Plasma CRH levels were measured in patients with non-insulin dependent diabetes mellitus (NIDDM) as the pituitary-adrenal abnormalities have been reported in NIDDM. They were also measured after oral administration of 75g glucose to examine whether glucose increased plasma CRH along with insulin secretion. The baseline plasma CRH was significantly lower in diabetic patients than in controls. Baseline ACTH and cortisol were significantly higher in NIDDM patients than in controls. Plasma CRH, ACTH and cortisol did not change after glucose administration in either NIDDM patients or controls. Neither plasma CRH nor ACTH showed a significant correlation with plasma glucose or insulin response in NIDDM patients. These results suggest that CRH secretion is not stimulated by glucose, that plasma ACTH and cortisol are increased in NIDDM patients and that CRH is not responsible for these increases.

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We studied plasma polymorphonuclear elastase (PMNE) levels before and during methimazole treatment in patients with hyperthyroidism. Plasma PMNE levels in patients with hyperthyroidism were significantly higher than those of normal subjects and patients with thyroid adenoma. Plasma PMNE levels in patients with Graves' disease were significantly reduced when their thyroid functions after methimazole treatment were normal, compared with the levels of the patients before treatment. In patients with hyperthyroidism there was a significant positive correlation between plasma PMNE and serum FT3, but no correlation between plasma PMNE and serum TRAb. The results suggested that PMNE was secreted from blood neutrophil in hyperthyroid state patients with Graves' disease and that it was beneficial for helping determining peripheral thyroid function.

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The cytologic localization and cellular levels of insulin receptors in the human ovary during follicular growth, regression and atresia were examined by the avidin/biotin immunoperoxidase techniques with a monoclonal antibody to insulin receptor. In primordial follicles, only the oocyte showed a weak immunostaining for insulin receptor, whereas the stromal cells surrounding primordial follicles were moderately immunostained. The earliest stage of follicular growth at which immunostaining for insulin receptor in granulosa cells and theca interna cells became apparent was the preantral stage. With the increase in the size of the follicles, the immunostaining of the oocyte and follicular elements intensified, whereas the staining intensity of the stromal cells surrounding growing follicles was reduced compared to those surrounding primordial follicles. The immunostaining in granulosa and theca interna cells persisted in the corpus luteum, and further intensified during the midluteal phase. In the regressing corpus luteum, the immunostaining was present only in the peripheral lutein cells adjacent to the central scar tissue. The corpus albicans was negative for the immunostaining, but the surrounding stromal cells exhibited predominant staining. In atretic follicles, the theca interna cells exhibited intense staining for insulin receptor without appreciable staining in the scattered granulosa cells, whereas the surrounding stromal cells were moderately immunostained.
This is the first study to demonstrate notable changes in insulin receptor expression in the oocyte, granulosa cells, theca cells, lutein cells and surrounding stromal cells during follicular growth, regression and atresia. The results obtained indicate insulin participation in oocyte maturation, follicular growth and stromal cell function. The increased expression of insulin receptors in theca interna cells of atretic follicles and in stromal cells surrounding the corpora albicans raises the intriguing possibility of insulin involvement in the transformation of theca interna cells into stromal cells. This implies that insulin may participate in remodelling ovarian local tissues following follicular atresia and luteolysis in the human ovary.

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We have already reported that the serum of the mid-pregnant rat contains placental lactogen-α (PL-α), which has a higher molecular weight than PL-I when analysed by gel-filtration chromatography. In order to characterize PL-α, we prepared antipeptide antisera directed against the hydrophilic regions of authentic PLs and developed RIAs with them. With these RIAs, antiserum AI-86 reacted only with PL-I, whereas antiserum AII-86 reacted with both PL-α and PL-I, which indicates that the antibody recognition sites of these molecules are not identical and their core proteins differ distinctly. As the use of the RIAs with AI-86 and AII-86 in combination enabled PL-α and PL-I to be discriminated, the glycoresidues and hydrophobicity of PL-α were analyzed further. Analysis with hydrophobic chromatography revealed that PL-α was less hydrophobic than PL-I and, furthermore, PL-α, but not PL-I, bound specifically to a wheat germ agglutinin (WGA) lectin column. Therefore, in view of their different amino acid sequences and glycoresidues, PL-α and PL-I are distinct molecular entities.

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Plasma concentrations of three major vitamin D₃ metabolites, 25-hydroxyvitamin D₃ (25-OHD₃), 24R, 25-dihydroxyvitamin D₃ (24, 25-(OH)₂D₃), and 1α, 25-dihydroxyvitamin D₃ (1, 25- (OH)₂D₃), were determined serially in patients with gynecological malignancies treated by chemotherapy which included cisplatin. While 25-OHD₃ and 24, 25-(OH)₂D₃ levels did not change regularly, 1, 25-(OH)₂D₃ decreased significantly (50%) after only one or two courses of the treatment in all cases. In concert with declining 1, 25-(OH)₂D₃, the PTH levels increased two or three times. The results were explained by the impairment by cisplatin of mitochondrial function in proximal tubules in the kidney.

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