Seasonal changes in plasma levels of testosterone (T) and 5α-dihydrotestosterone (DTH) were examined in the adult male Japanese red-bellied newt. The concentration of plasma T increased significantly (p<0.05) from January to February and decreased drastically from April to July when it reached its minimum. After the summer decline, plasma T increase rapidly from July to October (p<0.05) and reached its maximum in October. Concerning the seasonal fluctuation of plasma DHT levels, three peaks were observed; one, the highest peak, appeared in the middle of May, and two lower peaks in early spring (Feb.-March) and autumn (Sept.-Oct.). In this report, the role of plasma T and DHT in the reproductive cycle is discussed.

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A case of a 14-year-old boy with optic hypoplasia and pituitary dwarfism was presented. Eye Examination showed typical hypoplasia of the left optic nerve and mild hypoplasia of the right optic nerve. Except for dwarfism and nystagmus, the appearance of the patient was not unusual. Computed tomography revealed an enlargement of the suprachiasmatic cistern, and the presence of the septum pellucidum. The pituitary function tests revealed complete deficiency in GH, and poor or intermediate secretion in ACTH, TSH, PRL, LH and FSH. Urine volume and specific gravity were normal. Therapy with human growth hormone has been successful. It was indicated that hypopituitarism was probably of hypothalamic origin and early recognition of the syndrome is important in view of the possibility of treating growth retardation in some blind, or near blind children.

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To observe the effect of sex hormones on RNA synthesis of androgen-dependent tumors, mice of DD/S strain bearing androgen-dependent mouse mammary tumor (SC 115) were treated with the hormones, and RNA polymerase activities in isolated nuclei of the tumor were measured. The activity of RNA polymerase I in SC 115 was diminished to approximately 60% of the control level by castration, and it was restored to the precastrated level 12hr after treatment with 0.2mg of testosterone. While the activity of RNA polymerase II in the tumor was scarcely influenced by castration, it reached 150% of the castrated level 24hr after administration of testosterone. In CS 1, a subline of SC 115 with partial loss of androgen dependency on growth, castration and testosterone treatment caused similar but less prominent effects on the activities of RNA polymerase. Administration of estradiol-17β to castrated animals also increased the activity of RNA polymerase I, but to a lesser degree than testosterone, in both SC 115 and CS 1. However, only SC 115 demonstrated an increase in RNA poly merase II after estrogen treatment.

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A solid phase radioimmunoassay for the detection of anti-insulin antibodies has been developed and evaluated. After coating round-bottomed wells of polyvinyl chloride with porcine monocomponent insulin, 50μl of serum from either insulintreated diabetics or non diabetic controls was added to the wells, followed by the addition of ¹²⁵I-protein A, and then the radioactivity of the bound ¹²⁵I-protein A on the wells was counted. This solid phase radioimmunoassay is very simple, does not require centrifugal steps and the results correlated well with those of previously published immune precipitation methods (r=0.68, p<0.01).
Our present method requires only 50μl of serum and 4hours for the assay, if the wells are coated with insulin in advance. Therefore, this procedure is thought to be suitable for clinical use.

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Normal estrous or diestrous female, ovariectomized female and estrogen plus progestin-primed, ovariectomized female mice of the ICR/JCL strain adopted a lordosislike posture (LLP) in response to manual stimulation of the dorsal rump when they stood at the initiation of barbiturate anesthesia. Normal and castrated male mice also adopted a LLP following manual stimulation at the commencement of barbiturate anesthesia. In contrast, approximately 60% of anesthetized female mice at day 14 of gestation showed LLP without any tactile stimulation. In these pregnant mice, manual stimulation never brought about any change in the LLP incidence at the initiation of barbiturate anesthesia.

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Diethylstilbestrol was continuously infused into pregnant mice through the tail vein. In male fetus exposed to diethylstilbestrol (DES) from days 15 to 19 of gestation, Leydig cells of the testis were found to be increased in number when examined at 3-60days of age. Mitotic rate in the interstitial tissue was elevated in 0-to 3-day-old mice exposed prenatally to DES. The ratio of the intertubular space to testis size was greater at 3days of age than at 1day in prenatally DES-exposed mice, while in the controls, the rate was lower at 3days than at 1day. Spermatogenic activity was lower in DES-exposed adult mice than in the controls, as shown by the spermatogenic index.

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Thirty-nine episodes of hyperglycemia and disturbance of acid-base equilibrium were classified according to the result of nitroprusside test for serum (or urine) ketones, serum electrolytes, glucose, lactate, β-hydroxybutyrate and arterial blood pH and gas analysis into the following 6groups;(1) diabetic ketoacidosis (DKA), (2) mild DKA, (3) DKA with mixed acid-base disturbance, (4) DKA with lactic acidosis, (5) lactic acidosis with mild ketonemia, (6) lactic acidosis. Their clinical manifestations, laboratory findings, insulin and i.v. fluid requirement in the early phase of therapy were surveyed and compared with those reported from Western countries. The fundamental problems of groups (1) to (4) were hyperglycemia and acid-base disturbance. Groups (5) anp (6) were characterised by underlying serious medical illness, accompanied by lactic acidosis and hyperglycemia. All patients in groups (1) to (4) recovered but 7 of 10 patients in groups (5) and (6) died within the first 7days. DKA with or without lactic acidosis and lactic acidosis with or without mild ketonemia appeared as two separate conditions from the standpoint of management and prognosis and were differentiated only by nitroprusside test for serum ketones. DKA with lactic acidosis and DKA without it could not be differentiated by routine blood chemistries and therapy for the two did not differ so that they were thought to be in the same spectrum of metabolic alteration.

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The present experiments were performed to elucidate the mechanism of the selection and maturation of a dominant follicle in women. Eight normally menstruating women undergoing myonectomy or tubal surgery volunteered for the present study. In 3 patients who were operated on Day 9-11, a visual dominant follicle was removed. The other 5 patients underwent the removal of a newly developing corpus luteum on Day 15-21. After taking 3 or 4 preoperative blood samples in the morning after their hospitalization, blood was obtained at 3 or 6h intervals for the first 36-45h and at 1-3day intervals thereafter for 21-34days. Serum FSH, LH, estradiol and progesterone were measured by radioimmunoassay.
Follicleectomy was followed by a sudden drop in estradiol and a minor increase in progesterone. FSH increased for a few days and then declined. There was a drastic, but short-term increase in LH following follicleectomy which was performed before a preovulatory gonadotropin surge. A LH surge occured 10.7±1.2days (mean ±S.E.) after follicular ablation followed by a luteal phase.
In contrast, there was no remarkable LH release in 4 out of 5 patients who underwent luteectomy. A slightly higher level of FSH was sustained for 2-7 postoperative days. “Luteal phase” rises in estradiol and progesterone terminated promptly following luteectomy. A LH surge was observed 14.2±1.7days after surgery followed by a luteal phase. After either type of operation, a sustained increase in FSH was followed by a gradual increase in estradiol which preceded a gonadotropin surge.
These hormonal sequences resemble those seen in the normal follicular phase. The present data demonstrated that follicleectomy and luteectomy bring on some characteristic hormonal changes which may exert stimulatory or suppressive effects on the selection and maturation of a dominant follicle after the removal of a main ovarian cyclic structure culminating in ovulation at a certain interval.

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A double antibody radioimmunoassay (RIA) for the measurement of bullfrog basic gonadotropin (GTH) is described. The antiserum was raised against a highly purified bullfrog basic GTH. Bullfrog basic GTH-IV (pI=9.33) was used for radioiodination and as a reference standard. The dissociation constant of the binding of the antibody to bullfrog basic GTH-IV was 8.28×10^-12M. The standard curve was obtained with 0.1-25.6ng/ml of bullfrog basic GTH-IV. When bullfrog sera were assayed by this RIA system, the intra-assay coefficient of variation and inter-assay coefficient of variation were 4.45% and 5.6%, respectively. The accuracy of the assay was shown as the recovery of bullfrog basic GTH-IV added to the pooled bullfrog sera. With a range of 0.1-12.8ng/ml, the recovery was 104.78±11.91%(Mean±S. D.). When bullfrog pituitary extract was fractionated by isoelectric focusing (IEF) and assayed for gonadotropin by this RIA system and also by radioreceptor assay systems employing Xenopus or Anolis testicular homogenates, the RIA positive peaks agreed well with those obtained with Xenopus RRA, indicating that the present RIA system measures LH-like gonadotropin. The inhibition curves of pituitary preparations from bullfrog, brown frogs and dark-colored frog were almost parallel to the standard curve, while those of other anuran and urodele species were not. This RIA system was highly sensitive, but allowed only a narrow application, because of species-specificity.

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Daily injection of 8IU of methionyl human growth hormone (met-hGH), produced by recombinant DNA technology, was performed in healthy young volunteers. Daily injections for four days did not cause any significant change in the results of physical examination, electrocardiogram, blood count or urinalysis.
Non-esterified fatty acid levels increased significantly at 4hours after the first injection of met-hGH in a manner similar to pituitary human growth hormone (pithGH). Both hGHs caused a significant increase in serum somatomedin A during the four daily injections. Plasma hGH reached peak levels at 3-4hours after im injection of met-hGH and 4-6hours after pit-hGH. The levels were 19.5±1.6 and 21.3±2.8ng/ml, respectively. After iv one bolus administration of 4IU hGH to two volunteers, plasma hGH reached peak levels at 5minutes at mean levels of 292ng/ml after methGH and 293ng/ml after pit-hGH. The half life of the two hGHs was 17.2 and 17.1 minutes, respectively, according to the disappearance curve measured after iv administration.

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A patient with Cushing's disease was described who showed non-suppressibilities of plasma ACTH, serum cortisol, and urinary 17-hydroxycorticosteroid (17-OHCS) with a standard dexamethasone suppression test. Serum cortisol was not suppressed even after a larger dose of oral and intravenous dexamethasone (up to 33mg). An intravenous injection of cortisol also failed to suppress the plasma ACTH level. Repeated dexamethasone suppression tests gave similar results. Since the patient had no adrenal tumor but a submucosal tumor of the stomach, ectopic ACTH syndrome was suspected. However, removal of the gastric tumor resulted in no improvement of hypercortisolism. Selective sampling from the petrosal vein showed a higher ACTH concentration than that in the peripheral vein. In addition, the salivary cortisol concentration after 8mg of dexamethasone administration was seen to decrease to 24.7% of the basal level, which was greater than that of serum cortisol and urinary 17-OHCS (65.4 and 78.0%, respectively). She underwent Hardy's operation, and adenomatous tissue 2mm in diameter was found and curetted out. However, hypercortisolism recurred after the transient remission. Additional treatments with medical adrenalectomy, pituitary irradiation, and administration of reserpine were carried out. After that, her clinical course was uneventful. We concluded that the determination of salivary cortisol was a reliable tool for evaluation of the response in dexamethasone non-suppressible Cushing's disease.

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Glucocorticoid therapy is not considered as an authentic method for obtaining euthyroid in Graves' disease. We tried the administration of prednisolone as a preoperative preparation for subtotal thyroidectomy in 4 hyperthyroid patients with Graves' disease who had suffered adverse effects of thionamide antithyroid drugs, including agranulocytosis, liver damage and skin eruptions.
Following oral administration of a 30 mg daily dose of prednisolone, with or without other antithyroid reagents, both serum T₄ and T₃ concentrations decreased rapidly and reached the normal range within 2 weeks. The clinical signs and symptoms of hyperthyroidism also improved rapidly and subtotal thyroidectomies were performed uneventfully in all cases.
These results suggest that 1) glucocorticoid medication can normalize the circulating hormone levels rapidly in Graves' disease, 2) it is a useful method as preoperative preparation for subtotal thyroidectomy, especially when other conventional methods are not available or effective in obtaining euthyroid, and 3) mechanisms other than thyroid stimulation by circulating immunoglobulin seem to play an important role in causing hyperfunction of the gland.

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Using 5 adult Japanese monkeys (Macaca fuscata) of both sexes, the qualitative and quantitative changes in plasma LH caused by LHRH administration were examined before and after castration. Five hundred μg synthetic LHRH was infused for 2hrs and blood samples were obtained every 15min for 5hrs under ketamine hydrochloride anesthesia. Plasma LH levels were determined by an in vitro bioassay using the mouse testis interstitial cell preparation. The LHRH infusion induced a biphasic release of in intact animals of both sexes and in castrated females but not in castrated males. The biphasic pattern of LH release was characterized by an initial acute rise followed a plateau or a fall and the consecutive second rise after a long latent period.
Aliquots of plasma samples obtained during 30-90 (corresponding to the initial rise) and 120-210min (corresponding to the second rise) after the initiation of LHRH infusion were pooled separately (plasma pools I and II, respectively) and subjected to isoelectrofocusing (IEF) fractionation. The IEF fractionation was performed in ampholine-sucrose gradient, pH range 3.5-10, under a constant electrical power supply (3W, 700V) for 48hrs at 4°C. After the completion of the IEF fractionation the LH activity in each fraction was measured by the in vitro bioassay. The IEF fractionation separated plasma LH into 6 distinct species in terms of pI values. When the IEF profiles of plasma pools I and II in intact animals of both sexes were compared, the increase and the decrease in acidic to neutral LH and alkaline LH, respectively, were consistent and significant in plasma pool II. Changes of the same kind were further exaggerated when the IEF profiles in castrated animals were compared with those in non-castrated ones.
The acidic LH may represent the LH species newly synthesized and not well processed. The LH species possessing pI values of 7.5-8.5 is likely to be the mature type of LH exhibiting high biological potency. The LH species migrating in the high alkaline region seems to be the LH of over-maturation.

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We have measured the population of Killer (K) lymphocytes in the peripheral blood of 108 patients with various kinds of thyroid disease. In the patients with Hashimoto's thyroiditis and Graves' disease, the relative and absolute numbers of K-lymphocytes were significantly lower than those seen in healthy controls (p<0.001), and the longer the duration of medication, the lower the K-lymphocyte population. However, there was no apparent correlation between the serum titers of thyroid autoantibodies and the K-lymphocyte population. In the patients with malignant and benign thyroid tumors, the relative and absolute numbers of K-lymphocytes significantly decreased when compared with those of controls (p<0.001), the decrease was more prominent after surgical operation than before operation. A prominent decrease in the K-l mphocyte population was evoked to maximum 2 to 4weeks after surgical operation. The patients with both malignant and benign tumors having abundant lymphocytic infiltration in their surgical specimens generally revealed a lower K-lymphocyte population than those having no lymphocytic infiltration.

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The role of central endogenous prostaglandin (PG) in the regulation of osmotically mediated vasopressin secretion under normal physiological conditions in rats was studied employing a prostaglandin synthetase inhibitor, indomethacin, and sensitive and specific radioimmunoassay for plasma arginine vasopressin.
A single dosage and doses for two and five days by s. c. injection of 5.0mg/kg BW of indomethacin, which can produce significant depletion of endogenous PG in the hypothalamus, resulted in a high correlation (r>0.9) between plasma arginine vasopressin (pAVP) and plasma osmolality (pOSM). This relation could be fitted to straight lines of the form pAVP=2.1 (pOSM-296)(N=10), pAVP=3.2 (pOSM-298)(N=13) and pAVP=0.9 (pOSM-295.6)(N=10), after i. p.injection with hypertonic saline, for the three dosages, respectively. The control regression lines could be described by relations of the form pAVP=2.9 (pOSM-298)(N=11), pAVP=3.1 (pOSM-299)(N=8) and pAVP=1.1 (pOSM-296.9)(N=13), respectively. The slope and intercepts (on the osmolality axis) were thus comparable with those of the control group, which had not been influenced by plasma urea nitrogen, glucose or blood volume.
These results suggest that the administration of indomethacin did not influence the osmoregulation of AVP directly.
From this evidence, we conclude that central endogenous PG has no primary role in the control of the osmoregulation of arginine vasopressin secretion under normal physiological conditions.

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Attempts were made to elucidate the possible antigonadotropic action of PRL at the ovarian level. Immature female rats of Wistar strain were injected subcutaneously with PMS (5-10i. u.) at 0900h on day 23 of age, followed an intraperitoneal injection of HCG (5-10 i. u.) at 1400h on day 25. In all animals treated with PMS-HCG, ova were found in the oviducts when examined at 0900h on day 26 of age. Rat PRL was administered either at various doses (1.0-20μg/day) in the morning (0800h) between day 23-25 or at 5μg/day for 10 days prior to HCG injection. The ability of exogenous gonadotropins to induce ovulation and weight gain of the ovary were not affected by this PRL treatment. In animals injected with PMS, significant rises in serum estradiol-17β levels were observed 48h later. This PMS-induced increase in the estrogen concentration was also not affected by the treatment with various doses of PRL. From these results, it seems likely that exogenously administered PRL may be unable to suppress the ovarian responsiveness to gonadotropins by direct action on the ovary.

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Attempts were made to find out whether hyperprolactinemia has an effect on the hypothalamo-pituitary response to estrogen feedback and LHRH stimulation. Adult female rats of Wistar strain were ovariectomized and received subcutaneous injection of 20μg estradiol benzoate (EB) 3-4weeks later (day-0). A second injection of 20μg EB, when administered at noon on day-3, induced a highly significant increase in serum LH (p<0.001 vs. basal values), but not FSH, estimated at 1800h on the same day. This EB-promoted LH release was not altered by pretreatment with rat PRL (5μg/day), which was administered subcutaneously daily in the morning (1100h) between day-1 and day-3. No statistical difference in the serum LH concentration was found when compared with the values for the control animals pretreated with 0.9% saline alone.
Serum gonadotropins 15min after LHRH administration (100ng/100g BW) in 32-day-old female rats were not statistically different between the animals pretreated with 5μg PRL, which was given subcutaneously daily (at 0800h) for 3days, and the controls pretreated with 0.9% saline. These results suggest that an acute increase in serum PRL may not exert a negative effect on the gonadotropin release induced by estrogen feedback and LHRH stimulation.

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