In order to clarify some of the developmental processes of the human adrenal cortex or steroidogenesis in infancy and childhood, serum concentrations of 17-hydroxypregnenolone, 17-hydroxypregnenolone sulfate and 17-hydroxyprogesterone were measured by means of a combined radioimmunoassay method, and the age-related changes in these steroids were also examined. The actual ranges of serum concentrations of 17-hydroxypregnenolone, 17-hydroxypregnenolone sulfate and 17-hydroxyprogesterone in umbilical cord blood were 27.1-80.5, 1, 560-5, 030 and 53.3-304nmol/l, respectively. These values subsequently decreased to nadirs of 0.95-2.09nmol/l of 17-hydroxypregnenolone in subjects 1 to 2 years old, 0.93-7.03nmool/l of 17-hydroxypregnenolone sulfate in subjects 3 to 6 years old and 0.18-0.78nmol/l of 17-hydroxyprogesterone in subjects 1 to 2 years old, respectively, and they were followed by gradual increases to the adult levels.
This study thus revealed the age-related changes in 17-hydroxypregnenolone and its sulfate concentrations in infancy and childhood and indicated that, in the process in which the adrenal cortex was differentiated to the definitive form, the decrease in the activity of steroid sulfotransferase in infancy and childhood occurred more slowly than the increase in that of 3β-hydroxysteroid dehydrogenase.

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Nuclear binding sites of T₃ in human trophoblastic cells were biochemically characterized. Nuclei were isolated by a combination procedure with mild homogenization of the freshly obtained trophoblastic tissue aged term gestation, centrifugations and Triton X-100 treatment. The isolated nuclei were incubated with various concentrations of ¹²⁵I-T₃ at 20°C for 3h. The total number of T₃ binding sites per nucleus was approximately 650. The apparent association constant (Ka) was 6.0×10⁹M^-1. Nuclear proteins extracted from purified nuclei with 0.4M KCl were able to bind T₃ giving rise to nuclear thyroid hormone binding protein-T₃ complexes and they were precipitated with bovine IgG, as a carrier protein, by 12.5% polyethylene glycol. Binding was maximum in 3h incubation at 20°C or in 18h at 0°C, while it dropped quickly at 37°C. The binding characteristics were analyzed by Scatchard plots. In nuclear proteins obtained from 8 term placentae there was a single set of high affinitylow capacity T₃ binding sites with Ka of 7.0×10⁹M^-1. The capacity is about 62.7 fmol T₃/mg DNA. The binding sites were found to be speciffc for L-T3, while L-T4 was about 100-fold less effective, rT₃ ineffective, and D-T₃ and D-T₄ were roughly 1/8 and 1/5 as active as L-T₃ and L-T₄, respectively in displacing ¹²⁵I-T₃ from the binding sites. These data confirmed that human placenta is target organ of thyroid hormones; trophoblastic cells contain T₃ nuclear receptors which are biochemically similar to those isolated from liver, although the capacity is low.

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Evidence for pre-receptor, receptor and post-receptor glucagon defects was investigated in adipocytes from streptozotocin-diabetic rats. For this purpose male Wistar rats were injected by cardiac puncture with streptozotocin (65mg/Kg body-weight) or saline solution and sacrificed after 7 and 15 days of drug administration. Increased glucagon levels and increased glucagon degradation in serum together with a decrease in glucagon binding were found in both groups of diabetic rats. The decrease in glucagon binding was related to a decrease in the number of glucagon receptors/cell rather than to a change in receptor affinity. The lipolytic response of glucagon was increased. However, the ability of glucagon to increase basal or theophylline-stimulated cAMP accumulation in the incubation medium of adipocytes from diabetic rats was decreased. Such alterations could represent a counter-regulatory mechanism of the hyperglucagonemia detected in streptozotocin-diabetic rats.

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To assess the role of protein kinase C and cAMP on the calcitonininduced alteration of phosphate accumulation by renal tubular cells, the effects of phorbol esters, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), and DBcAMP on the phosphate accumulation in LLC-PK₁ cells were investigated. Calcitonin stimulated phosphate accumulation with a concomitant increase in cAMP production. Phorbol esters and 1-oleoyl-2-acetyl-glycerol, activators of protein kinase C, also stimulated the phosphate accumulation. Furthermore, H-7, an inhibitor of protein kinase C, inhibited a calcitonin-induced increase in phosphate accumulation significantly. Although DBcAMP by itself did not increase the phosphate accumulation, it enhanced the stimulatory effect of 12-0-tetradecanoyl phorbol-13-acetate on the phosphate accumulation. Accordingly, protein kinase C as well as cAMP might be involved in the calcitonin-induced increase in phosphate accumulation in LLC-PK₁ cells.

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Painless (silent) thyroiditis (PT) occurred simultaneously in 1 male and 4 females aged 21 to 52 years working at a nursery school. Clinical symptoms did not include goiter, or pain or tenderness of the neck in any of the patients but were characterized by edema of the lower legs as well as palpitation and loss of body weight as observed in subacute thyroiditis. General blood analysis showed that all patients were negative for C-reactive protein (CRP) and 3 had mild impairment of liver function. Examination of thyroid function suggested transient thyrotoxicosis accompanied by a marked reduction in radioactive iodine uptake (RAIU), but antithyroglobulin hemagglutination antibody (TGHA) and antithyroid microsomal hemagglutination antibody (MCHA) were negative in all patients. Examination of various viral antibodies showed no significant changes in their titers. Thyrotoxicosis was transient and disappeared without treatment or by glucocorticoid administration. Our results suggested that PT observed in this study was caused by some environmental factor. The possibility of an unidentified virus as a factor cannot be ruled out.

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Histocompatibility lymphocytic antigen (HLA) typing was performed in 6 patients with acute exacerbation of Hashimoto's thyroiditis whose diagnoses were established on the basis of typical histological findings, and was compared with those of 12 with subacute thyroiditis, 33 with general Hashimoto's thyroiditis and also with a control group.
There was a high incidence of BW35 in patients with subacute thyroiditis, although it was only seen in 1 of 6 patients with acute exacerbation. The difference was statistically significant (p<0.01). Four of 6 patients with acute exacerbation had DR2 and none of them had DR4, which was the reverse of the findings for Hashimoto's thyroiditis patients in general, and the difference in the incidence of DR2 was significant (p<0.001). None of the HLA types in patients with acute exacerbation was significantly different from those of the control group.
In conclusion, HLA typing in patients with acute exacerbation was different from those of subacute thyroiditis and general Hashimoto's thyroiditis. Acute exacerbation was considered to involve quite a limited and rather unique population among patients with Hashimoto's thyroiditis.

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For purification of androgen receptor from hypertrophic human prostate, solutions used for elution of androgen receptor from DNA Sepharose, affinity labeling of the receptor and ability of affinity gel to retain the receptor were examined. Elution with 20mM pyridoxal 5'-phosphate of the receptor from DNA Sepharose was more efficient than that with diluted pyridoxal 5'-phosphate, high ionic solution or various concentrations of Mg⁺⁺. ³H-dihydrotestosterone bromoacetate was applicable to covalent binding with partially purified androgen receptor regardless of the low specificity of the ligand. Affinity gel of thiopropyl-Sepharose 6B coupled to 17α-(2', 3'-epoxypropyl)-5α-dihydrotestosterone was better than Affigel 102 coupled to N-[3-(3-oxo-5α-androstane-17β-yloxycarbonyl) propionyloxy] succimide or aminoethyl-Sepharose 4B coupled to 17α-carboxyethynyl testosterone with respect to the rate of retention of androgen receptor. In view of these observations, the following purification procedures were constructed: Removal of DNA Sepharosebinders from the cytosol, 40% ammonium sulfate precipitation, affinity chromatography using thiopropyl-Sepharose 6B coupled to 17α-(2', 3'-epoxypropyl)-5α-dihydrotestosterone, and DNA Sepharose chromatography. After affinity labeling of the receptor thus obtained, the molecular weight was estimated. Some 1300-fold purification with a yield of 0.25% of the androgen receptor was achieved. The molecular weight of the receptor was mainly 45 K with 90 K in a lesser amount. The Stokes radius was calculated as 30Å.

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Diurnal changes in vasopressin and oxytocin levels in cerebrospinal fluid were investigated under normal diurnal conditions. The patients examined had ruptured intracranial aneurysms, and underwent neck-clipping operations and continuous drainage from the basal cistern. All of the patients recovered consciousness without signs of neurological deficit. The investigations were conducted for 2 days starting 5-9 days after the neck-clipping operations were performed. The oxytocin concentration decreased as night fell, remained low during this period and then increased during the day. The vasopressin level demonstrated no definite rhythmic tendency. No correlation was revealed between the changes in the concentrations of either vasopressin or oxytocin in the cerebrospinal fluid and the osmolality.

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It has been reported that melatonin produces either progonadal or anti-gonadal effects in mammls, depending on the time and mode of administra-tion. Information on he melatonin-effect on the testis in toads indicated varied changes during the breeding and hibernating seasons.
The present study in Rana hexadactyla (Lesson), a continuous breeder revealed that administration of melatonin at a dosage of 50μg/frog/day either in the morning or evening for a week inhibited spermatogenesis; however, when melatonin was administered for a longer period, this inhibitory effect was lost. Moreover, treatment with melatonin both in the morning and evening had no net effect on the testes.

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A 21-year-old woman with Cushing's syndrome was found to have a marked diurnal variation in cortisol secretion. Serum cortisol concentrations and urinary excretion of 17-hydroxycorticosteroids were normal in the morning but clearly increased in the afternoon. The patient was cured by resection of an adrenocortical adenoma. ACTH and prostaglandin E1 stimulated cortisol release from incubated adenoma tissues in vitro. The cause of the abnormal diurnal rhythm of cortisol secretion is unknown.

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Renal resistance to antidiuretic hormone (ADH) has been speculated to be a mechanism of transient nephrogenic diabetes insipidus occurring during late pregnancy. In order to study possible involvement of ovarian steroids in this mechanism, their effect on cyclic adenosine 3': 5'-monophosphate (cAMP) response to arginine vasopressin (AVP) was examined utilizing rat and human renal medullary cells in monolayer culture. In both rat and human cells, estradiol significantly reduced cAMP response to AVP; estradiol at 1.84×10^-8M, 1.84×10^-7M and 1.84×10^-6M decreased cAMP production stimulated by 10^-8M AVP to 78±5%, 67±2%(P<0.05) ana 52±1%(P<0.001) of the control in rat renal cells, respectively, and in human renal cellsthe effect of estradiol was comparable to that in rat cells. In rat renal cells progesterone also reduced cAMP response to AVP dose-dependently; progesterone at 1.59×10^-7M, 1.59×10^-6M and 1.59×10^-5M decreased cAMP production stimulated by 10^-8M AVP to 87±1%, 72±5%(P<0.001) and 37±5%(P<0.001) of the control, respectively. On the other hand, corticosterone and dexamethasone at concentrations ranging from 10^-8M to 10^-5M and aldosterone at concentrations ranging from 10^-9M to 10^-5M did not alter cAMP response to AVP significantly. The suppressive effect of estradiol increased with time until six hours and thereafter it reached a plateau. Simultaneous addition of estradiol and progesterone resulted in a cumulative suppressive effect. Thus, both estradiol and progesterone reducecd renal cAMP response to AVP. It is suggested that these ovarian steroids themselves may reduce the renal responsiveness to AVP when they increase, for example in late pregnancy.

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Purification of porcine thyroid TSH receptors was attempted by using Lubrol, TSH-Sepharose, gel filtration and the immunoabsorption by anti-thyroglobulin (Tg) antibody-Sepharose. The effects of microbial protease inhibitors on the degradation of TSH receptors were also evaluated.
Porcine thyroid membrane was solubilized with 1% Lubrol-PX and TSH receptors were purified by TSH affinity chromatography, followed by gel filtration on TSKgel-G3000SW. A partial purification was achieved; a 692- fold purification and 49.3% recovery of high affinity binding site and a 409- fold purification and 29.1% recovery of low affinity binding site, respectively.Rechromatography on G3000SW gel and immunoabsorption by anti-Tg antibody-Sepahrose resulted in further reduction of contaminants. Among 10 proteaseinhibitors studied, a low concentration of chymostatin was the only one thatshowed a significant protective effect on the degradation of the receptors (p<0.05).

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The aim of this study is to examine the influence of Sertoli cells on LH bindingto Leydig cells in culture in immature mice. Leydig cells and Sertolicells were obtained from the testes of immature C57BL/6Ncrj mice and were cultured in serum-free medium for 7 days. The LH binding to Leydig cells and the FSH binding to Sertoli cells were dependent on incubation time, the number of cells, and the amount of labelled hormone added. The dissociation constant for LH binding to Leydig cells was 7.3×10^-10M. Co-culture of Leydig cells with Sertoli cells for 7 days decreased LH binding to Leydig cells.The binding was 34.9% of that to Leydig cells cultured alone. After cultivationof Leydig cells with spent Sertoli cell-cultured medium (SM) for the last 4 days of the 7-day culture period, LH binding to Leydig cells decreased to as low as 17.4% of that of the controls. For the controls, LH binding was measured in Leydig cells cultured in spent Leydig cell-cultured medium (LM). There was no difference between SM-and LM-cultures in the final survival rate or the percentage of cells showing histochemically demonstrated 3β-hydroxysteroid dehydrogenase activity. These data suggest that some factor or factors are secreted from the cultured Sertoli cells and inhibit the bindingofLH to Leydig cells in culture.

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Alterations in catecholamine (CA) and thyroid hormone metabolism were examined in a 12-year-old girl with anorexia nervosa during 3 months of treatment. Accordingto her body weight change, the observation period was divided into 3 stages: initial emaciation (stage 1). stable maintenance of the -30% level of the previous weight (stage 2) and convalescent stage (stage 3). Stage 1 was characterized by relatively high urinary norepinephrine (NE) andepinephrine (E) but low dopamine (DA) excretion, elevated plasma DA-βAhydroxylase (DBH) activity and reduced serum thyroid hormones, especially the triiodothyronine (T3) level. In stage 2, urinary CAs were markedly suppressed, while serum thyroid hormones gradually increase. Instage 3, a great increase in DA excretion, a fall in plasma DBH activity and normalization of thyroid hormones were observed. In the low T3 state below 60ng/dl, urinary NE+E/DA ratios were elevated and widely fluctuated (0.58±0.30, SD), but were gradually decreased and completely stabilized in the normal T3 state (0.07±0.02, P<0.001). These results indicate that (1) although total CA production was depressed in anorexia nervosa, a change from an adrenergicdominantto a dopaminergic-dominant state occurs in accordance with body weight gain, and (2) this shift in the CA profile is associated with concomitantrecovery of reduced thyroid hormone concentrations. Thus, as for the energy expenditure, compensatory changes were observed in CAs and thyroid hormones in relation to caloric restriction.

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To evaluate the prevalence of hyperprolactinemia in patients with polycystic ovarian syndrome (PCO), 72 patients with oligo-or anovulation were studied. All of the patients had persisting elevated LH (>25mIU/ml), normal FSH, high LH/FSH ratio (>2.5), and exaggerated LH responses to LHRH. Mean testosterone and androstenedione concentrations were appreciably increased in these patients. Out of 171 samples for prolactin (PRL) determination from these 72 patients, only 5 patientshad a PRL value above 30ng/ml during the first sampling. The next sampling fromthese same 5 women disclosed that they were transiently hyperprolactinemicbecause the next samples showed a normal PRL value. To further investigate the PRL secretory capacity 500μg of TRH and 10mg of metoclopramide (MCP) were administered to these 72 and 44 patients, respectively. The PRL response to MCP was significantly blunted in these patients compared to normal women while the PRL response to TRH in these patients was not indistinguishable from that in normal women.These results indicate that the true prevalence rate of hyperprolactinemia inPCOmay be low rather than high and the association of hyperprolactinemia with PCO may be coincidental rather than a pathogenically related phenomenon.

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A 34-yr-old woman with hypertension (142/102mmHg), hypokalemia, high plasma and urinary aldosterone and low plasma renin activity was studied. A left adrenal tumor and enlarged right adrenal gland were demonstrated by adrenal venography. During administration of dexamethasone (2mg daily, for 3 weeks), urinary aldosterone excretion decreased abruptly from 22.5 to 9-11μg/day, serum potassium increased and blood pressure fell to 120-130/80-90 mmHg. After left adrenalectomy, all manifestations improved with no medication. The resected adrenal gland revealed clear cell adenoma and micronodular adrenocortical hyperplasia. The patient was considered to be a rare case of glucocorticoid-suppressible hyperaldosteronism with an aldosteroneproducing adenoma.

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Anterior pituitaries were removed from female rats at various stages duringthe estrous cycle and FSH was fractionated by isoelectric focusing (IEF). Fourteen FSH components were observed during the estrous cycle and twelve of them were distributed between pH 3.71 and 6.66. IEF profiles of FSH in the pituitaries varied with the stage in the estrous cycle. Especially at the time of serum FSH surge on the day of proestrus, most of the components decreased, while only a highly alkaline component showed an increase. When these FSH components were separated and their nature was examined by radioreceptor assay (RRA), radioimmunoassay (RIA) and gel filtration, differences were observed among these components in the RRA/RIA, ratio and gel filtration profile. As a general tendency, the RRA/RIA ratio of the components became greater while the apparent molecular size became smaller, as their pI became higher. However, some highly acidic components showed a biphasic elution pattern and the most acidic one eluted the latest on gel filtration, suggesting that these components may be heterogeneous in terms of molecular size. The FSH concentration in sera collected at different stages in the estrous cycle was measured by both RRA and RIA. The RRA/RIA ratio was high when the serum immunoreactive FSH was low, and low during the FSH surge.
From these findings, it is concluded that the quality of FSH molecules present in the anterior pituitary gland changes dynamically throughout the estrous cycle, especially during the period of serum FSH surge. Furthermore it is suggested that the type of FSH secreted from it also varies according to the stage in the estrous cycle.

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We studied the effects of shakuyaku-kanzo-toh (a Chinese herbal medicine) and its components on testosterone production by rat gonads. We used paeoniflorin as a main component of shakuyaku (paeoniae radix), glycyrrhizin as a main component of kanzo (glycyrrhizae radix) and glycyrrhetinic acid as a main metabolite of glycyrrhizin. Oral administration of shakuyaku-kanzotoh, glycyrrhizin, and glycyrrhetinic acid decreased in vitro basal testosterone production in Leydig cells by LH stimulation. Glycyrrhizin and glycyrrhetinic acid caused a significant decrease in testosterone production with an accumulation of 17α-hydroxyprogesterone when incubated with isolated Leydig cells, while paeoniflorin showed no such effect. The inhibitory effect of glycyrrhetinic acid was far more potent than that of glycyrrhizin, causing about 90% inhibition at 10μg/ml. Glycyrrhizin and glycyrrhetinic acid did not change the cyclic AMP or progesterone level in the Leydig cells. When ¹⁴C-labeled androstenedione was incubated with microsomal fraction of testicular or ovarian tissue, glycyrrhizin and glycyrrhetinic acid inhibited the conversion of androstenedione to testosterone, indicating that these compounds inhibit the activity of 17β-hydroxysteroid dehydrogenase (EC. 1. 1. 1. 64). The ED⁵⁰ of glycyrrhetinic acid was about 4 μM.

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To examine a possible role of atrial natriuretic peptide (ANP) in water and electrolyte disturbances associated with thyroid disorders, plasma ANP levels were studied in patients with hyper- and hypothyroidism. In 5 of the 21 hyperthyroid patients, including two patients with atrial fibrillation and two patients with mild cardiomegaly, the plasma ANP concentration was increased when compared to normal subjects. After treatment with methimazole or propylthiouracil, the plasma ANP concentration fell to normal in 4 patients, while it remained high in one patient who had persistent atrial fibrillation. No significant correlation was found between plasma ANP and the heart rate in untreated hyperthyroid patients. Plasma ANP was within the normal range in all 8 patients with hypothyroidism. During treatment with T₄, the plasma ANP concentration increased in 6 of the 7 patients. Chest X-ray films and ultrasonic echocardiography demonstrated pericardial effusion in 4 of these patients before therapy. A weak but significant correlation was found between the plasma ANP and T₄ concentration, and between plasma ANP and free T₄ in hyper- and hypothyroid patients before and after treatment. These results indicate that abnormalities in ANP dynamics in thyroid disorders may probably be caused by hemodynamic changes resulting from a thyroid hormone excess or deficiency.

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Developmental changes in the pituitary responsiveness and the secretory pattern of FSH and LH in response to a single injection of LH-RH (100ng/rat, s.c.) as estimated by increases in plasma concentrations of FSH and LH 10, 30 and 60min after the injection were studied in female rats at 5, 10, 15, 20, 25 and 30 days of age. The pituitary responsiveness to LH-RH for both FSH and LH release increased from 5 to 15 days of age, reached a maximum on 15 days of age and declined thereafter, whereas a marked increase in the amount of these hormones in the pituitary occurred between 15 and 20 days of age. An apparent change in the secretory pattern of both FSH and LH was observed from 20 days of age onward. In groups up to 15 days of age, plasma concentrations of FSH and LH remained elevated 60 min after the injection of LH-RH, though the plasma concentration of these hormones returned to preinjection concentrations in groups at 20 days of age or later. These results indicate that the age-related changes in the secretory pattern of LH and FSH in response to LH-RH as well as changes in the pituitary responsiveness were apparent during the prepubertal period.

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