Gonadotropin and steroid hormone levels in both peripheral and ovarian venous blood were measured in samples obtained from 20 Japanese patients with polycystic ovary syndrome (PCOs) and 10 normal women in early follicular phase (normal women) by radioimmunoassay. The change in the amount of steroid hormone following intravenous human menopausal gonadotropin (HMG) or dexamethasone administration was investigated. The mean concentration in patients with PCOs was significantly higher than the concentrations found in normal women for LH (p<0.001), but not for FSH in peripheral blood. Significantly elevated ovarian venous steroid hormone levels in PCOs were found for 17α-hydroxypregnenolone (p<0.05), progesterone (p<0.05), 17α-hydroxyprogesterone (p<0.01), ⁴Δ-androstenedione (p<0.01), testosterone (p<0.01), estrone (p<0.01) and estradiol (p<0.05), but not for dehydroepiandrosteronesulfate (DHEAS). The ovarian dehydroepiandrosterone (DHEA) level was slightly elevated in PCOs. The concentration of ovarian ⁴Δ-androstenedione in PCOs reached twelve times as much as that in normal women. After the administration of HMG, all of the ovarian venous steroid hormone levels were elevated slightly and without significance in the short observation time for 10min. The DHEAS level was suppressed while the ovarian DHEA level remained high in PCOs following dexamethasone administration. These findings seem to indicate there is no adrenal involvement and no adrenal-like component in the ovary of PCOs, and no evidence of 3β-hydroxysteroid dehydrogenase and/or aromatase deficiency in this study. The increase in the steroid hormone secretion in PCOs is explained by the increase in ovarian production in polycystic enlarged ovaries.

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Immunohistochemical characterization of the human pituitary β(R) cells was investigated through the findings of the immunoreactivities with anti-porcine ACTH, -rat TSH, -rat LHβ and -rat FSH sera. Immunostained corticotrophs are oval or round in shape and localized in the anteromedial wedge. It is shown on the adjacent sections that they correspond to the β(R) cells with amphophilic stainability with PAS-iron hematoxylin. In this wedge, amphophilic cells are preponderant, but PASpositive thyrotrophs and gonadotrophs are not numerous. Amphophilic stainability varies in degree from cell to cell: One cell contains numerous medium-size of secretory granules weakly stained with iron hematoxylin and strongly with PAS in the PAS-positive cytoplasm, and the other cell is filled with big secretory granules intensively stained with iron hematoxylin and weakly with PAS. The immunostained TSH, LH and FSH cells are different from the β(R) corticotrophs, because anti ACTH serum never reacts to the TSH, LH and FSH cells in the two adjacent sections. LH and FSH reactivities are observed in the single cells. It is concluded that human corticotrophs are amphophilic β(R) cells filled with secretory granules, and that they have quite a different appearance from the rat chromphobic stellate corticotrophs with a row arrangement of secretory granules along the plasma membrane.

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Hyperinsulinemic and hypoglycemic rats with islet cell tumors induced by streptozotocin were shown to have significantly decreased immunoreactive insulin and somatostatin concentration (expressed as per g wet weight (w. wt)) and content (expressed as per whole pancreas) in the pancreas surrounding the tumor as compared to those of age and weight adjusted controls. The pancreatic glucagon concentration of the former was significantly lower than that of the latter but no significant difference between glucagon content in the two groups was observed. Thus, inverse relationships were demonstrated between circulating insulin and somatostatin as well as insulin in the pancreas. It is suggested that a large amount of insulin in plasma results in a decrease in pancreatic insulin and somatostatin.

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Intraventricular administration of tonin, an enzyme which forms angiotensin II directly from a natural protein substrate (angiotensinogen), stimulated water drinking and increased systemic blood pressure in rats. These responses were abolished by the simultaneous administration of an angiotensin II analogue, but unaffected by an angiotensin I-converting enzyme inhibitor. We confirmed, in vitro, that angiotensin II was generated when the brain homogenate was incubated with tonin in the presence of DFP and EDTA. These results indicate that the central effects of tonin on water drinking and systemic blood pressure are mediated via the direct formation of angiotensin II in the brain, and suggest the possibility that the tonin-angiotensin II system in the central nervous system might be important in controlling water balance and blood pressure.

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During the postnatal development of rat pituitary it has been shown that PRL cells appear oval, polygonal or cup shaped in both sexes. The oval cells first appear on Day 10 after birth. Development of the latter two shapes is delayed to Days 20 and 30, respectively. The polygonal shaped cells become predominant with sexual maturation. Some of them surround the gonadotrophs with their elongate, cytoplasmic processes, taking on the appearance of cup cells. Although the PRL cells are more numerous in the female than in the male, the cup shaped cells are inverse. Further it is noted that the immunostainability of the oval and polygonal cells varies during the estrous cycle; the weakest reaction was revealed at estrus and the strongest at dioestrus. Those immunohistochemical findings may suggest that the oval cells are premature, the polygonal ones mature and the cup ones particularly differentiated. The topographic affinity between the cup cells and the gonadotrophs may be a morphological indication of their intimate functional relationship.

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The effect of Bordetella pertussis vaccine on plasma glucose, insulin and glucagon secretion was investigated in normal and alloxan dogs. On the 8th day after the vaccine injection, in normal and alloxan dogs during the infusion of arginine and glucose, the plasma glucose level was lower and the IRI level was higher than in the saline controls. On the other hand, the plasma IRG level showed no significant difference between the vaccine and saline control groups of normal dogs, but in alloxan dogs this vaccine made the plasma IRG level lower during arginine infusion than in the saline controls and suppressed it significantly during glucose infusion.These effects of the vaccine disappeared on the 30th day after its injection into normal and alloxan dogs.
It is suggested that in normal dogs Bordetella pertussis vaccine decreased plasma glucose through the promotion of insulin secretion without any effect on glucagon secretion, while in alloxan dogs this vaccine might alleviate hyperglycemia through the enhancement of insulin and the inhibition of glucagon secretion.

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Antisera to an LH-RH analogue, des-Gly¹⁰-[D-Leu⁶]-LH-RH-ethylamide (TAP-144) were produced in 10 rabbits. By using these antisera, a specific and sensitive radioimmunoassay for TAP-144 was established. Sensitivity of the radioimmunoassay ranged from 5 to 100 pg per assay tube with these antisera. LH, FSH, TRH, LH-RH, and the 1-6 fragment of TAP-144 did not practically cross-react with these antisera. However, the antisera showed a tendency to cross-react with LH-RH analogues, though the degree of the cross-reactivity differed among individual sera. The average crossreactivity of the 10 antisera showed low specificities to TAP-144 analogues which are altered at positions 2, 3, 4, 5, and 6, but showed high specificities to those altered at positions 8 and 9. The antisera also showed low cross-reactivities to LH-RH analogues replaced at position 6 by D-amino acids and those altered at position 10 by alkylamines, but they showed fairly high cross-reactivities to analogues which are altered simultaneously at both positions 6 and 10.
When TAP-144 was administered intraperitoneally to rats on the diestrous day, serum concentrations of TAP-144 increased dose-dependently but maximal serum concentrations of both LH and FSH were attained in response to higher doses of TAP-144. The peak LH and FSH concentrations appeared 70 to 110 min after the peak TAP-144 concentration had been reached. Similar delays in reaching the peak LH and FSH levels were also observed when TAP-144 was administered intravenously, subcutaneously and intramuscularly. When TAP-144 was administered intravaginally, a low but constant serum level of TAP-144 was maintained from 5 to 300 min after the administration, but serum LH and FSH levels declined to a low level from 180 min after the TAP-144 administration.

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PMSG-induced prepubertal pregnant rats were injected with hCG on Day 7 of pregnancy to induce the formation of secondary corpora lutea (CL) in addition to the primary CL of pregnancy. The rats bearing two sets of CL were hypophysectomized on Day 11 of pregnancy (Day 3 of the secondary CL age) and their primary CL were removed on Day 13 of pregnancy (Day 5 of the secondary CL age). The operations did not affect the fetal growth until Day 18 of pregnancy when the weight of and progesterone secretion of the secondary CL rapidly increased. Hysterectomy on Day 18 of pregnancy (Day 10 of the secondary CL age), however, induced regression of CL and rapidly decreased the progesterone secretion. These results suggest that placentas between Days 15 and 18 of pregnancy could maintain the function of CL at Days 7-10 of CL age when LH is indispensable as a luteotropic hormone, and that placentas between Days 18 and 21 of pregnancy could support the rapid growth of CL at Days 10-13 of CL age. Placental hormones which would participate in these luteotropic events have been discussed.

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The movement of subcellular calcium in the bile pool of hepatocyte was investigated after a single intraperitoneal administration of calcium chloride in rats. The administration of calcium (4.0 mg/100 g BW) produced a remarkable elevation of serum calcium and a corresponding increase in liver calcium. The calcium taken by the liver cells at 10 min after calcium administration was markedly located into the nuclei, mitochondria and microsomes, and this distribution was not accompanied by a significant elevation in the cytosol level. At 20 min after calcium administration, the calcium increase above the subcellular structure was clearly reduced. On the other hand, serum calcium was markedly increased by calcium administration in both intact and thyroparathyroidectomized rats. However, the liver calcium increase induced in intact rats by calcium administration was much more than that in thyroparathyroidectomized rats. Also, the bile calcium level was markedly elevated by calcium administration in intact rats, but no elevation was observed in thyroparathyroidectomized rats. The present results suggest that the calcium taken by the liver cells is bound to the nuclei, mitochondria and microsomes, and then transported into the biliary duct.

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A 23-year-old man with primary hyperparathyroidism which was typical except for reduced alkaline phosphatase activity is reported. Histological examination of surgical specimens revealed chief cell hyperplasia of the parathyroid glands. Systemic abnormalities of alkaline phosphatase were demonstrated, i. e., marked reduction of all isoenzymes and undetectable osseous enzyme in the serum, abnormal distribution of the enzyme in hepatocytes and diminished enzyme activities in leukocytes. In addition, diminished bone remodeling activity was revealed in a biopsy specimen of the rib. The association of hypophosphatasia is highly unlikely, because of normal urinary excretion of phosphoethanolamine, lack of osteomalacia, and no indication of an hereditary factor. The causal relationship between low remodeling activity and abnormalities in alkaline phosphatase was suggested.

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The effect of TSH (100mU/ml) and norepinephrine (100μM) on the cyclic AMP levels was studied in 10 human normal tissues, 10 thyroid adenomas and 4 thyroid carcinomas (3 papillary and 1 follicular). Normal tissues responded to TSH with a marked elevation of the cyclic AMP level. Response patterns of 10 thyroid adenomas to TSH were variable ; the patterns of 6 cases resembled those of normal tissues, 3 responded mildly, and one had no response to TSH. Thyroid carcinomas had a higher basal level of cyclic AMP than those of normal tissues, although they responded only slightly to TSH. Two among 4 thyroid carcinomas had no response to TSH. Norepinephrine stimulated the accumulation of cyclic AMP in 4 thyroid adenomas and 3 thyroid carcinomas, while it had little effect on normal tissues. Responses to norepinephrine was observed only in thyroid tumors, although they had low response to TSH. It is suggested from these results that tumor cells originating from thyroid follicular cells have a modified response to hormones due to neoplastic alterations.

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Rat hepatoma cells (R117-21B) metabolized 3, 3'-diiodothyronine and 3'-monoiodothyronine by sulfation and glucuronidation. Glucuronidation of iodothyronines (a mixture of 3, 3'-diiodothyronine, -90%, and 3'-monoiodothyronine, -10%) was very rapid and 83% of the total ¹²⁵I in the medium was their glucuronides after 8hr incubation with the cells at 37°C. Iodothyronine sulfates (-9% of the total ¹²⁵I) were also detected. No clear increase in free iodide was observed during the 8 hr incubation.
The effect of KCN or dinitrophenol on glucuronidation of thyroxine was investigated. Both drugs inhibited glucuronidation, and dinitrophenol was more effective than KCN. Dinitrophenol accelerated the liberation of 1251-in the metabolism of thyroxine in the cultured rat hepatoma cells.

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Testosterone and 5α-dihydrotestosterone (DHT) concentrations in tissue and body fluid were determined in adult male rats to investigate the mode of androgen distribution.
The highest concentration of androgen (testosterone +DHT) and the smallest ratio of DHT/testosterone were observed in the intratesticular tissue fluid. The androgen concentration in testicular vein blood was one third of that in the intratesticular tissue fluid, but this concentration was much higher than in any other sample of body fluid. Among the body fluid samples, except for the intrascrotal fluid, DHT/testosterone ratio remained almost equal to that in the intratesticular tissue fluid. The intrascrotal fluid and blood samples from the testicular and epididymal arteries had significantly higher androgen concentrations than thoracic duct lymph or aortic blood.
Among the tissues, the accessory sex organs and kidney fat pad had higher androgen concentrations and larger DHT/testosterone ratio than muscle, including the levator ani muscle. The DHT/testosterone ratios in muscle specimens were not different from those in body fluid. The caput epididymis and surrounding adipose tissue (the epididymal fat pad) had the highest androgen concentration, and the largest DHT/testosterone ratio was observed in the caput epididymis.
These observations support the theory of the local transport of androgen from the testis to the caput epididymis and from the testicular vein to the testicular artery. Thus, the intrascrotal androgen targets, especially the caput epididymis, receive androgen effectively, and it was suggested that the epididymal fat pad plays an important role in maintaining a high androgen concentration in the intrascrotal androgen targets.

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The effect of thyroid hormone on the serum level of somatomedin A was studied using a radioreceptor assay for somatomedin A. In 13 patients with hyperthyroidism, the mean level of somatomedin A was 0.60±0.05U/ml, which was within the normal range. Low levels were found in 19 patients with primary hypothyroidism with a mean of 0.39±0.04 U/ml. After treating these patients with thyroxine, the serum somatomedin A levels returned to the normal range. There was a significant correlation between the somatomedin A level on one hand and T₃ (r=0.49) and T₄ (r=0.55) level on the other.
The serum level of somatosmedin A did not decrease for 7 days after thyroidectomy nor increase after treating hypophysectomized rats with 2 μg of thyroxine for 14 days. However, the effect of growth hormone on serum somatomedin A was augmented 42.2% by the combined administration of the same amount of T₄ for 7 days. There was a positive correlation between the serum level of somatomedin A and the percentage increase in body weight (r=0.73). These data suggest that thyroxine potentiates the effect of GH on synthesis and/or the secretion of somatome din A.

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Plasma level of norethindrone (NET) and in vivo release of norethindrone acetate (NETA) were studied in three groups of albino rabbits (5 animals/group) after insertion of one, two and four subcutaneous implants, each containing 40 mg crystalline NETA over a period of 24 weeks. The ratios of the in vivo release rate of steroid were 1, 1.9 and 3.9 in the animals of group 1 (one implant), group II (two implants) and group III (four implants) respectively. Thus, the in vivo release rate in group II and III showed an increase which was almost twice and four times as great as that of group I. However, the mean ratios of the serum NET levels were 1, 1.2 and 2.4 in animals of group I, II and III respectively. Thus, interestingly, the serum NET level did not show the expected twofold and fourfold increase and lacked correlation with the in vivo release. Although the insertion of multiple implants gives multiple increases in the in vivo steroid release, it does not give rise to a multiple increase in the serum levels of the steroid. It is possible that there is a kind of a threshold of steroid concentration in the animals when they are loaded with exogenously administered steroid. When the steroid concentration tends to cross the threshold level, pharmacokinetic or pharmacodynamic processes of the animal work maximum to hold down the steroid level in blood plasma.

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The relative potency in the hypothalamic-pituitary-adrenal (HPA) suppression of both prednisolone and betamethasone was examined in an acute study with normal volunteers and in a chronic study with glucocorticoid-treated patients. Circadian rhythm of plasma cortisol was studied after a single dose administration of 5 to 30 mg prednisolone or 0.5 to 3.0 mg betamethasone at 08:00 hr. Morning-rise of plasma cortisol occurred on the morning after the administration of 30 mg or less prednisolone but no morning rise was noted after the administration of 1.0 mg or more betamethasone. Plasma ACTH was slightly elevated on the morning after 30 mg prednisolone administration but showed low levels throughout the night after 3.0 mg betamethasone administration. Plasma cortisol responsiveness to ACTH was examined in patients before and during therapy with either prednisolone or betamethasone. The basal cortisol level was not suppressed and the responsivenessto ACTH remained nearly normal during long-term 5 mg prednisolone therapy, but these were completely suppressed during long-term 0.5 mg betamethasone therapy. The responsiveness to ACTH was nearly normal in patients receiving alternate-day therapy with prednisolone in such large doses as 50 or 60 mg every other day, but was completely suppressed in patients receiving 1.0 mg betamethasone every other day. The relative potency of betamethasone in acute and chronic suppressive effects on the HPA system seems to be much stronger than that of prednisolone inequivalent doses with comparable anti-inflammatory effects. It is also suggested that the alternate-day therapy with such long-acting steroids as betamethasone are useless in preventing HPA suppression.

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The amount of specific [³H]-progesterone bindings was measured in cytosols and in nuclear fractions of the hen hypothalamus and pituitary during the ovulatory cycle. In both tissues, a decrease in cytosol binding from the peak at 18 and 8 hr before ovulation was coincident with an increase in nuclear binding which attained a peak 14 and 6 hr before ovulation. The results suggest that progesterone may act on the hypothalamus and pituitary 18-14 hr before ovulation and 8-6 hr before ovulation.

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We have recently reported that high-fiber diet is an effective therapeutic regimen for the control of some metabolic derangements in diabetic rats (Yamashita et al., 1980). Additional effects of high-fiber diet on the metabolic states in streptozotocininduced diabetic rats were investigated. The animals were divided into two groups: a dietary fiber group fed on diet containing 10% dietary fiber ad libitum and a control group fed on a control diet ad libitum for fifty days. Fasting blood glucose levels were significantly lower in rats in the dietary fiber group than in those in the control group. In addition, plasma high density lipoprotein (HDL) cholesterol values and the HDL cholesterol/total cholesterol ratio in fiber-fed rats were higher than those in the control group. These results indicate that a high-fiber diet may have a beneficial effect on the metabolism of cholesterol as well as glucose in diabetics.

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