In order to investigate the action point of intraphysiological or supraphysiological elevation of FSH during the preovulatory period on follicular development, adult guinea pigs underwent unilateral ovariectomy on days 10, 12 and 14 of the estrous cycle (N=6 each group). Thereafter, guinea pigs were injected twice daily with either vehicle or pregnant mare's serum gonadotropin (PMS). After 2 days, the remaining ovaries were removed. The resected ovaries were fixed, embedded in paraffin, serially sectioned (7μm) and stained with Azan. All follicles >70μm were classified by size and atretic stage. The follicular size distribution was not affected by hemicastration at day 10, although the ratio of atretic follicles (>400μm) decreased from 51% to 32%(P<0.01). Hemicastration at day 12 increased the largest nonatretic population (70-99μm group) from 17% to 26%, and the ratio of atretic follicles (>400μm) decreased from 35% to 23%. The peak size distribution of follicles was shifted from 70-99μm to 200-299μm by PMS, and follicles 600-899μm in size contained an increased percentage of atresia, which resulted in the bimodal distribution of viable follicles >400μm. These data suggest that 2 day hemicastration promotes an influx of primordial follicles into growing follicles and suppresses the atretic process by a different mechanism depending on the date of hemicastration in the estrous cycle. Conversely, hemicastration+PMS accelerated viable follicle growth to increase the percentage of atresia.

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The effects of bilateral lesions of the hypothalamic paraventricular nuclei (PVN), of rats with a mean weight of 260g body, on eating habits and body weight, as well as on sympathetic nervous system (SNS) activity in interscapular brown adipose tissue (IBAT) were investigated. In 59 of 131 Sprague-Dawley female rats, PVN lesions resulted in hyperphagia and obesity. Although lesions were considered successful when more than 50% of the PVN was destroyed histologically, such lesions were observed in 35.9%(47/131) of all lesioned rats and all of these 47 rats were obese. Therefore, in this study, these 47 rats which were confirmed histologically, were designated as “PVNlesioned rats”. Plasma insulin levels in these 47 PVN-lesioned rats were more than double those of the controls. However, no significant differences were observed between plasma glucose levels in PVN-lesioned and control groups. Norepinephrine turnover, a reliable indicator of SNS activity, in IBAT, heart and pancreas was similar in PVN-lesioned and sham-operated control animals, even under contrasting conditions of feeding (ad libitum and fasting) and temperature (22°C and 4°C).
It is concluded that PVN lesions produce hyperphagia, obesity and hyperinsulinemia in rats with an average body weight of 260g without affecting the SNS activity in IBAT, heart or pancreas.

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Pregnancy is thought to create a metabolic condition of accelerated starvation. To clarify this idea, the effect of fasting on pregnant rats (day 21 of gestation) and their fetuses was examined. Although pregnancy significantly increased plasma insulin, plasma ketone body concentrations in fed pregnant rats were higher than those of age-matched fed virgin rats. After 48 hr fasting (i. e., fasting during days 19-21 of gestation), plasma insulin, was markedly decreased in virgin rats compared with term pregnant rats, while ketone bodies were significantly higher in pregnant rats than in virgin rats. Body weight was lower in fetuses from fasted mothers than those from fed mothers. Starvation also markedly diminished the insulin response to glucose in isolated, perfused pancreases in both virgin and pregnant rats. The amount of insulin released during glucose stimulation was greater in pregnancy, and the inhibitory effect of 48 hr fasting on insulin release was greater in virgin rats than in pregnant rats. It is possible, therefore, that in term pregnant rats a decrease in insulin release caused by fasting may cause more profound catabolism than in nongravid rats.

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Epidermal growth factor (EGF) is known to stimulate proliferation of various mammalian cells and secretion of prolactin (PRL) from rat anterior pituitary tumor cells. The effect of an acute increase in serum PRL induced by thyrotropin releasing hormone (TRH) or metoclopramide (MCP) on the serum immunoreactive EGF concentration was examined in nine hyperprolactinemic patients and eight normoprolactinemic women. The basal level of serum EGF in normoprolactinemic subjects was 472.8±51.1 pg/ml (Mean±SEM), which was not signifiantly different from that in hyperprolactinemic patients (487.8±22.5pg/ml). The serum EGF concentration was decreased to 40-50% of the basal level after the abrupt increase in serum PRL induced by the injection of TRH or MCP in normoprolactinemic subjects, but no significant change in serum EGF occurred in hyperprolactinemic patlents atter MCP injection, in spite of a significant increase in PRL. These results suggest that an acute increase of serum PRL in normoprolactinemic women, but not in hyperprolactinemic patients, suppresses serum EGF.

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Cyclic replacement therapy using estrogen and progesterone was instituted in 28 patients with gonadal dysgenesis and 13 patients with hypopituitarism.
When estriol was given at a dose of 2mg per day, 10 patients (9 gonadal dysgenesis and 1 hypopituitarism) developed hyperreninemia and 3 of the 10 patients (all gonadal dysgenesis) were associated with hypertension.
These side effects subsided within 6 months when the therapy was discontinued or the dose of estriol was decreased to 1 mg per day in addition to β-blocker.
Hypercholesterolemia was observed in 8 patients, but not related to high blood pressure.
Attention should be paid to plasma renin activity and blood pressure when estrogen and progesterone are given for the development of genitalia in patients with gonadal dysgenesis.

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We administered either saline (group A) or 10 IU of pregnant mare serum gonadotropin (PMS; groups B and C) to female immature rats. Fifty-three hours later, the rats were injected with saline (groups A and B) or 30 IU of human chorionic gonadotropin (hCG; group C). The rats were decapitated 17 h after the last treatment, and the serum levels of progesterone (P₄) and estradiol (E₂) were measured by specific radioimmunoassays (RIA). The receptor levels of progesterone (PR) and estrogen (ER) in the uterus and ovaries were measured and the dissociation constant (Kd) of PR was obtained.
The highest serum level of P₄ was found in group C and that of E₂ in group B. Cytsol levels of PR and ER in the uterus and ovary of the group B were the highest. It was indicated that the PMS treated-group (B), which had developing follicles in the ovary and the high serum level of E₂, showed the highest concentration of ER and PR in both the ovary and the uterus. In the PMS and hCG-treated group (C), the uterine and ovarian steroid receptors decreased probably because of the luteinization and the high serum level of P₄. The Kd uterine PR value was less than that of ovarian PR.

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The correlation between response of plasma GH to GHRH and the GHRHinduced stimulation of the intracellular adenylate cyclase (AC) activity in pituitary adenoma cell membranes in acromegalic patients was investigated. Each peak plasma GH level after iv administration of GHRH ranged from 1.1 to 13.8 times the basal level in 13 acromegalic patients. On the other hand, the maximal stimulation of intracellular AC activity (cAMP production) induced by GHRH varied from 1.4 to 6.4 times the control level in each GH-producing pituitary adenoma cell membrane. A significant positive correlation (r=0.89, P<0.005) between plasma GH response to GHRH and intracellular cAMP production stimulated by GHRH was observed in nine of the acromegalic patients. In contrast, the response of plasma GH to GHRH was significantly blunted, despite a fairly large production of intracellular cAMP stimulated by GHRH, in the other four acromegalic patients. These results suggest that GHRH-induced GH release from GH-producing pituitary adenomas of patients with acromegaly may be regulated not only by GHRH receptor-adenylate cyclase system but also modified by several other factors including somatostatin and Sm-C.

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We investigated the effects of sodium depletion and orthostasis on the plasma concentration and urinary excretion of vasopressin (AVP) in eight normal female subjects. After 4 days on a sodium controlled diet (130 mEq/ day), the subjects were placed on a low sodium diet (30 mEq/day) for 3 days and 120mg of furosemide was administered orally on the first day of the low sodium regimen.
Sodium depletion in the present study reduced body weight by 1.6 kg and increased hematocrit by 3.5%.
A significant (p<0.05) increase in plasma AVP and a significant (p<0.05) decrease in 24-h urinary excretion of AVP were observed during sodium depletion.
One-hour ambulation significantly increased plasma AVP in both control and sodium depleted phases (p<0.01). The percent change in plasma AVP tended to correlate with that in mean blood pressure in the control phase (r=0.69, 0.05<p<0.1), and significantly correlated in the sodium depleted phase (r=0.86, p<0.01).
The present results suggest that AVP may play an important role in the maintenance of blood pressure during orthostasis in the sodium depleted state.

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Eight-week-old, female Wistar fatty rats and their lean littermates were given a 30% sucrose solution in addition to a laboratory chow diet and water for 7 weeks. The fatty rats were hyperinsulinemic and hyperlipidemic, but normoglycemic when they drank only water. The hepatic activities of insulininducible glucokinase (GK), pyruvate kinase (PK), and malic enzyme (ME) were higher in the fatty rats than in the lean rats, whereas the insulin-suppressible glucose-6-phosphatase (G6Pase) activity was similar in both types of rats, indicating the normal response of hepatic enzymes to hyperinsulinemia in the fatty rats.
When they drank the sucrose solution, the fatty rats, but not the lean rats, developed hyperglycemia over 200mg/dl. Plasma insulin and triglyceride concentrations increased in both types of rats. Although the hepatic activities of GK, PK, and ME in the lean rats, and PK and ME in the fatty rats increased in response to the increase in plasma insulin, GK activity decreased in the fatty rats. On the other hand, G6Pase activity increased in both types of rats. As a result, the G6Pase/GK ratio, which may reflect net glucose handling in the liver, increased twofold in the fatty rats, but did not alter in the lean rats.
From these findings, we conclude that sucrose ingestion induces an increase in hepatic glucose production through derangement of the hepatic enzyme profile and that the resultant decrease in hepatic glucose handling may be one of the pathogenic factors participating in the development of hyperglycemia in Wistar fatty rats.

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A total of 80 patients with Turner's syndrome were treated with methioninefree recombinant hGH (r-hGH) for one year. Thirty-nine patients were treated with r-hGH at weekly dosage of 0.5IU/kg and forty-one were treated with 1.0 IU/kg/w by daily sc injection. Both treatment groups showed a statistically significant growth increase during the treatment from 3.7±1.0 to 6.0±1.1 and 7.2±1.3 (Mean±SD) cm/year, respectively. Fifty-nine percent of the patients treated with 0.5 IU/kg/w and 87.8% of the patients treated with 1.0 IU/kg/w showed a growth rate more than 2cm greater than the pretreatment values. Plasma somatomedin C levels were elevated and no remarkable advances in bone age were observed during the treatment in both treatment groups. An antibody against to hGH was observed in 6.8% of the patients. Otherwise, there were no significant changes in physical or laboratory examinations. No glucose intolerance was observed.
These results indicate that hGH treatment is useful in accelerating growth velocity in patients with Turner's syndrome.

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Ten clomiphene-resistant, normoprolactinemic women with polycystic ovary syndrome (PCOS) were treated by continuous and increasing administration of bromocriptine (Brc), and changes in hormonal profiles as well as therapeutic efficacy were examined. Ovulation was restored in four patients (responders), and two of them became pregnant, whereas the other six patients did not ovulate with Brc alone (non-responders). Endocrine analyses revealed distinct differences between responders and non-responders: 1) pretreatment serum levels of dehydroepiandrosterone sulfate (DHAS) in non-responders were significantly higher than those in responders: 2) exaggerated LH secretion was definitely aggravated with Brc therapy in non-responders, but unchanged or slightly reduced in responders: 3) basal PRL secretions showed a marked reduction in both groups, whereas this response to TRH in responders decreased more markedly than in non-responders with the therapy.
It is concluded that low DHAS group patients of PCOS are likely to respond to Brc, whereas high DHAS group patients appear contraindicated for this treatment due to its aggravating effect on LH secretion.

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A 42-year-old woman with Cushing's disease was endocrinologically cured after transsphenoidal selective excision of a left-sided microadenoma. Simultaneous samplings from the bilateral inferior petrosal sinuses with ovine corticotrophin-releasing factor (CRF) stimulation were performed preoperatively to evaluate some relations between the localization of a possible microadenoma and the levels of ACTH in inferior petrosal sinuses. The data for the venous samplings were as follows:-(1) The basal levels of ACTH obtained simultaneously from both inferior petrosal sinuses and peripheral vein were about the same.(2) A significant difference, with a high level of ACTH from the ipsilateral side of the microadenoma, was demonstrated after CRF stimulation.(3) The magnitude of increase in ACTH on the contralateral side after CRF was similar to that of the peripheral response.
It is recommended that all patients with ACTH dependent Cushing's syndrome and negative radiological findings should have bilateral simultaneous inferior petrosal venous sampling with CRF stimulation.

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We have clearly demonstrated corticotropin-releasing hormone (CRH) immunoreactive cell bodies and nerve fibers in the human hypothalamus by immunocytochemistry using free-floating sections instead of paraffin-embedded sections. Human hypothalami were obtained at autopsy, fixed and cryostatsectioned at 40μm. Free-floating sections were immunostained with antibody to CRH using the Vector ABC system. Most of CRH immunoreactive nerve fibers ffom the paraventricular nucleus pass under the fbrnix, while some CRH immunoreactive nerve fibers pass beyond the fornix and some through the fornix. Then the CRH immunoreactive nerve fibers run downward, medially to the supraoptic nucleus and toward the pituitary stalk.
This method of immunocytochemistry is a very sensitive and suitable means for immunocytochemical studies of neuropeptides in the human brain.

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To investigate the role of parathyroid function in transient hypocalcemia after subtotal thyroidectomy for Graves' disease, the serum parathyroid hormone (PTH) concentration and nephrogenous (N) cAMP were measured in 16 patients before and after surgery. Serum PTH was measured with two commercially available kits (PTH-M, PTH-C), PTH-M is a recently developed highly sensitive assay using an antibody recognizing the mid-portion of human PTH and a synthetic ¹²⁵I-tyr⁴⁵-human PTH (43-68) as a radioligand. One of the 16 patients had severe clinical tetany and had a markedly lower PTH-M concentration and NcAMP after thyroidectomy. However, no significant change in serum PTH-M, PTH-C and NcAMP were observed in the other patients, although their serum calcium (Ca) concentrations decreased significantly.
The Data were analyzed by dividing the patients according to the change in serum Ca or PTH. Serum PTH-M and PTH-C significantly decreased in 4 patients whose serum Ca clearly decreased after surgery. Serum Ca on the first postoperative day was significantly lower in patients whose serum PTH decreased after thyroidectomy than in patients whose serum PTH did not. Furthermore, the serum Ca concentration was significantly correlated with PTH-M, and with NcAMP on the third postoperative day. These data proved that hypofunction of the parathyroid gland is important in transient hypocalcemia after subtotal thyroidectomy for Graves' disease. The pathogenetic mechanism of transient hypocalcemia was discussed in comparison with the data from a patient who had overt parathyroid injury.

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Rat epidermal growth factor (rEGF) was isolated from the submaxillary gland of male rat by reversed phase high performance liquid chromatography and ion exchange chromatography. The binding of purified rEGF to human carcinoma cells (A-431) and its tritiated thymidine uptake on rat epidermal fibroblast cells (FR) were almost the same as those of purified commercially available mouse EGF (mEGF). Antisera to rEGF was raised in rabbits and a radioimmunoassay (RIA) system was established. The assay range of the RIA was about 1.0 to 100ng/ml. The within assay coefficients of variation were 5 to 10%, while the between assay coefficients of variation were 5 to 13%. The tissue content of rEGF of male rats (10 weeks old) was examined. As a result, the submaxillary gland was found to contain a very high concentration of rEGF (214μg/g wet tissue) as predicted, and digestive tissues, stomach, intestine and duodenum contained 2.49, 3.57, 9.44ng/g wet tissue, respectively. The amounts in prostate and seminal vesicle were relatively high, being 65.6 and 2, 268ng/g wet tissue, respectively. The amount in the submaxillary gland increased markedly after 7 weeks of age. These results suggest that EGF is an important factor in gonadal function.

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The management of hyperthyroidism due to Graves' disease in Japan was the subject of a survey of the members of the Japan Thyroid Association (JTA), and the results were compared to those of the European Thyroid Association (ETA). In the questionnaire, in vivo and in vitro diagnostic procedures, the choice ot treatment and the details of the treatment for a patient with typical, moderate and uncomplicated hyperthyroidism due to Graves' disease was at first asked, and eight variations with a single alternative were proposed to evaluate how each alternative would affect the choice of treatment. For the diagnostic procedures, thyroid uptake/scintigraphy was carried out by approximately 60% of the respondents and the isotope mainly used was ¹²³I. The number of in vitro tests used for diagnosis averaged 8.1±1.8 tests. Measurements of basal TSH and free T₄ were the most frequent tests performed to confirm the diagnosis of hyperthyroidism (94 and 80%, respectively). Determinations of microsomal, thyroglobulin and TSH-receptor autoantibodies were also employed by many respondents (96, 96 and 77%, respectively). On the other hand, the free T₄ index and TRH test were less frequently employed. In the teatment of these patients, antithyroid drug treatment was the first choice, and surgery was not, in general, regarded as a primary therapy except in a patient with a large goiter. The frequency of the respondents who advocated radioiodine therapy was considerably higher for patients with recurrences and old age. No respondents proposed radioiodine therapy for young patients. Specialists tended to favor their own specialist treatment regimens. The initial dose of antithyroid drugs was reduced according to thyroid function, and withdrawal of antithyroid drug treatment was determined by some specific criteria (basal TSH in supersensitive assays, TSH-receptor autoantibodies, T₃ suppression test, etc.). The aim of radioiodine therapy and surgery was to restore euthyroidism. The significant differences between the results from the JTA and those from the ETA were as follows radionuclide used for thyroid uptake/scintigraphy was mainly ¹²³I in Japan, but ¹³¹I in Europe, the number of diagnostic studies in Japan was more than that in Europe, and the dosage of antithyroid drugs was reduced according to thyroid function and discontinued based on certain specific criteria in Japan, but after fixed periods in Europe. These results may represent actual trends in how hyperthyrodism due to Graves' disease is managed in specialist clinics in Japan today and the differences between the JTA and the ETA.

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We present a report on two sisters who have 17α-hydroxylase deficiency with hyperaldosteronism. They have hypertension and hypergonadotropic hypogonadism. The steroid profiles suggest that they have 17α-hydroxylase deficiency. In contrast to the classical biochemical findings in 17α-hydroxylase deficiency, both of these patients have hyperaldosteronism. Thus this report describes a new variant of 17α-hydroxylase deficiency with hyperaldosteronism.
Dexamethasone suppressed the mineralocorticoid excess, including aldosterone, and improved their hypertension. In the untreated state, ACTH, instead of the renin-angiotensin system, regulated plasma aldosterone levels, but during dexamethasone treatment the renin-angiotensin system regulated these levels.

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