A radioimmunoassay specific for the amino (N) terminal portion of the parathyroid hormone (PTH) molecule (N-PTH radioimmunoassay) has been developed by iodinating synthetic ^1-34bovine PTH (^1-34bPTH) and using commercially available bPTH antiserum. A radioimmunoassay specific for the carboxyl (C) terminal (C-PTH radioimmunoassay) has been carried out by adding enough amount of 1-34bPTH to the PTH radioimmunoassay system, The data obtained from N-and C-PTH radioimmunoassay were compared with those obtained from the PTH radioimmunoassay. It was observed that plasma levels of N-PTH, indicating biologically active PTH, were only one 8th to 32th to those of PTH and those of C-PTH were almost equal to those of PTH. These data corresponded well with those reported previously by using the antiserum specific for each terminal of the PTH molecule from the other laboratory. The half life of plasma N-PTH and C-PTH determined following the removal of parathyroid adenoma was less than 10min and about 45min respectively. These data indicate that the N-PTH radioimmunoassay can be done by iodinating ^1-34bPTH and using commercially available antiserum.

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Time course of the changes in insulin release and cyclic AMP levels in isolated rat islets incubated in media containing 5 or 16.7mM of glucose were followed.
The higher glucose concentration caused a slight but significant increase of cyclic AMP levels after 10min incubation, but not 5min incubation, whereas the stimulation of insulin release by 16.7mM of glucose was apparent in both incubation times.
Theophylline increased cyclic AMP levels markedly but did not stimulate insulin release when the glucose concentration was 5mM. A slight augmentation by theophylline of insulin release was observed in the incubation medium containing 16.7mM glucose.
All these findings suggest that the elevation of cyclic AMP in islets may not play a role for the initiation of the insulin release induced by glucose, though it may act to modulate the glucose effect.

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An epidemiological survey on the incidence of juvenile chronic lymphocytic thyroiditis was performed in 10, 220 apparently healthy school children in Ishikawa district, Japan. The subject of present study included 6, 244 school children (2, 831 boys and 3, 413 girls, ages 6-18 yrs.) in Kanazawa City and 3, 976 children (2, 055 boys and 1, 921 girls, ages 6-18 yrs.) in Wajima City. The first group was selected as a representative of urban area and the second group as that of seaside area.
Children who have goiter or firm thyroid were selected for testing antithyroglobulin and anti-microsomal antibodies in sera. Final diagnosis of chronic lymphocytic thyroiditis was made on histological specimen obtained by needle biopsy on the antibody positive subjects.
The overall incidence of chronic lymphocytic thyroiditis in these children was 3.0 per 1, 000, whereas the incidence in adolescent girls was as high as 8. 2per 1, 000. There was a considerable sex difference in the prevalence, the ratio of female to male was 6.5: 1, and the incidence increased with age. The incidence in seaside area was 5.3 per 1, 000 that was significantly higher than in urban area, I.4 per 1, 000 (p<0.005).
Histologically, 26 of 30 cases (87%) were classified as focal thyroiditis and 4 cases (13%) were diffuse thyroiditis.
Serum T₄-I and T₃ values were within normal range in all patients, but resting TSH was elevated in 1 of 23 cases and TSH response to TRH was exaggerated in 3 of 23 cases. Impaired organification of iodide was observed in 6 of 32 cases by iodide-perchlorate discharge test.
The present study demonstrates that juvenile chronic lymphocytic thyroiditis is highly prevalent among apparently healthy school children and early recongnition of the disease with preventive care for hypothyroidism in future should be stressed.

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This study was undertaken to evaluate the relative contribution of insulin, proinsulin-like components (PLC) and C-peptide toward plasma levels of immunoreactive insulin (IRI) and C-peptide immunoreactivity (CPR) in the pig and to elucidate the mode of secretion of PLC in the early phase of insulin release.
Following the intravenous glucose loads, the concomitant secretion of CPR with that of IRI occured rapidly and the maximum plasma level of IRI was observed at an earlier time than that of CPR. Following the intravenous glucagon injection, the maximum plasma levels of IRI and CPR were observed at the same time in the early phase. After the gel filtration of acid alcohol extracts of plasma in a fasted state, a very small amount of PLC and a small amount of C-peptide as well as a small amount of insulin were detected. The results obtained from the gel filtration of extracts revealed that the increased amounts in IRI and CPR after the injection of glucose or glucagon consisted mostly and respectively of insulin and C-peptide in the pig, because the concentration of PLC increased only slightly in the early phase. In fact, plasma levels of CPR and IRI were essentially and respectively paralleled to those of insulin and C-peptide which were assayed after the gel filtration of extracts. In addition, the slight elevation of PLC in the early phase after these stimulations indicated that PLC was elicited into blood circulation at the same time of the secretion of insulin and C-peptide.

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Twelve of 14 female mice of the ICR strain which had received a single injection of 50μg estradiol-17β on day 17 of fetal life exhibited irreversible cornification or stratification of the vaginal epithelium which persisted after ovariectomy until sacrifice performed 42-48 days later. Eight of the 12 mice had corpora lutea in their ovaries removed at 3-5 months of age. A similar injection of estradiol on day 15 of fetal life induced irreversible cornification or stratification of the vaginal epithelium in 6 of 12 females and only one of the 6 had corpora lutea in its ovaries when removed at 3-5 months. Mice given the same dose of estradiol on the day of birth or at 3 days of postnatal age invariably had ovaries bearing follicles of varying sizes and hypertrophied interstitial tissue but no corpora lutea. Changes in the vaginal epithelium in these animals were less remarkable as compared to that in prenatally treated mice.

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The boiling method deviced in accordance with the extraction procedure of secretin was applied to the extraction of gut glucagon-like immunoreactive material (GLI) and compared with the acid alcohol method of Kenny with respect to efficiency of the extraction and property of the extracted materials. GLI was extracted from minced porcine small intestine by each method. The total amount of GLI extracted by the boiling method was 14.45±2.07μg/10g small intestine (mean±S. E.), showing a high yield as compared to 4.07±0.29μg/10g small intestine obtained by the acid alcohol method. The difference was statistically significant (p<0.005).
The gel chromatogram of the acid alcohol extract was separated into two peaks; peak I appeared before the insulin marker, while peak II was eluted with the glucagon marker. The chromatogram of the boiling extract has a main broad fraction including insulin marker and a minor second peak corresponding to peak II of the acid alcohol extract. Boiling of the acid alcohol extract did not cause any shift of peak I in chromatogram.
GLI present in the first half of the main fraction of the boiling extract was different from that in the latter half, but identical to peak I of the acid alcohol extract with respect to the immunoreactivity against glucagon antibody.
It is concluded that in the extraction of GLI not only high yield is achieved but one or more new components is picked up by the boiling procedure as compared to the acid alcohol method.

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Urine TRH was estimated by the radioimmunoassay which was accomplished according to the method of Bassiri and Utiger. Minimum detectable dose of TRH was 25 pg and recovery of TRH ranged from 72% to 112% in our laboratory. Intraassay coeffcients of variation were 5.4% to 14.0% and interassay variations were 10.6% to 15.0%. Of the TRH analogues tested, only two (Ser-His-Pro-NH₂, Thr-His-Pro-NH₂) had potent reactivity to anti-TRH serum in large dose of 100ng/tube. Urine samples were kept at-20°C after adjusted to pH 3.0 because the inactivation of TRH in urine was markedly dependent on temperature and pH value. Using this radioimmunoassay, diurnal variation of the urinary TRH excretion at regular intervals in normal subjects was observed. Peak TRH excretion occurred around early morning, while minimum of the excretion was observed around noon. Total urinary TRH excretion of 24 hours was 817-1579 ng (M±SE: 1241±89ng) in normal subjects. In patients with chronic renal failure, urinary excretions of TRH was obviously lower than those of normal subjects.

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The effects of infusion of glucosamine on immunoreactive glucagon (IRG) and insulin (IRI) secretion were studied in dogs and ducks. During systemic infusion of glucosamine, hyperglycemia developed and insulin secretion was inhibited in both species. An immediate and sustained elevation of peripheral IRG levels was induced in ducks but a transient rise, detectable only in the pancreatic vein blood, was provoked in dogs. Suppression of insulin release and stimulation of glucagon release may be mediated by the inhibition of glucose utilization in β-and α-cells. The very prompt response of IRG in ducks may imply that glucosamine has a specific stimulating effect on the α-cells of ducks. Intrapancreatic administration of glucosamine in dogs, however, failed to elicit the rise of IRG, although insulin secretion was inhibited. Thus, it is suggested that the systemic administration of glucosamine in dogs may stimulate IRG secretion by some indirect effect.
In one dog, however, a sustained rise of the pancreatic vein IRG was observed. Thus, the possibility cannot be ruled out that the difference in IRG response to glucosamine in dogs and ducks is quantitative rather than qualitative. Glucagon release by glucosamine may provide an additional factor to the hyperglycemic effect of glucosamine, in addition to its effect to suppress insulin release as well as its direct inhibitory effect on glucose utilization in tissues.

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A radioimmunoassay (RIA) method for luteinizing hormone (LH) releasing hormone (RH) utilizing rabbit antiserum against synthetic (Glu¹)-LH-RH coupled with human serum albumin at the N-terminus, is described. This assay system for LH-RH also cross-reacted with several LH-RH analogues or fragments, but not with pituitary trophic hormones. The assay was performed on the hypothalamic extracts of adult ovariectomized rats and female immature rats which had been treated with estradiol. The FSH and LH levels in the pituitary gland and serum of the same animals were determined by RIA. The radioimmunoreactive LH-RH content of the stalk median eminence markedly increased seven days after ovariectomy. The serum levels and the pituitary contents of FSH and LH of the same rats were also significantly augmented. In immature rats, the hypothalamic content of LH-RH, as measured by RIA, was significantly increased one hour after the injection of estradiol. The FSH and LH levels in the pituitary showed a significant rise after 7 hours.

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A and B cells of pancreatic islets of the dog and mouse were stained by the enzyme-antibody method. A cells, in general, were small in number, particularly in the mouse, and distributed at the peripheral zone of the islet. Furthermore, A cells were located very close to the intrainsular capillary. On the other hand, B cells occupied dominantly the islet and mainly existed in its central area, especially in the mouse and not adjacent to the intrainsular capillary.
A positively reacting cell with anti-insulin antibody was also found in the uncinate process of the dog pancreas, while there was no positively reacting cell with anti-glucagon antibody. This fact, as there were only two kinds of islet cells, B cell and the third cell of the islet (named D cell) in the uncinate process, showed that the D cell had neither glucagon nor glucagon-like immunoreactivity.

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Cytosol 9S receptor was prepared from the supernatant fluid at 105, 000×g of rat prostate homogenates by a Sephadex G-100 gel chromatography, and was labeled with ¹³¹I. The ¹³¹I-labeled 9S receptor retained the activity of forming a complex with ³H-dihydrotestosterone, similarly to the intact cytosol receptor, when examined by a sucrose density gradient centrifugation. When the ¹³¹I-labeled cytosol 9S receptor was incubated with isolated prostatic nuclei in the presence of ³H-dihydrotestosterone, it was found that the ¹³¹I-labeled receptor was directly incorporated into the nuclei in a form of the complex bound to ³H-dihydrotestosterone. ¹³¹I-Labeled receptor-³H-dihydrotestosterone complex which was incorporated into the nuclei was extracted with 0.5 M KCL solution, and the nuclear complex sedimented with a velocity of 5S. The incorporation of ¹³¹I-labeled receptor-³H-dihydrotestosterone complex into the nuclei increased along with the raised temperature of incubation, whereas association of the complex with the chromatin reached maximum at 35°C and then decreased gradually beyond this temperature. In the time course study, either the incorporation of the complex into the nuclei or the association with the chromatin reached the maximal levels within 10 min and leveled off up to 60 min. On the other hand, ¹³¹I-labeled serum protein was fhr less efficiently incorporated into the nuclei than the radioiodinated 9S receptor, and association of the serum protein with the chromatin was limited to a very little extent. The ¹³¹I-labeled 9S receptor-dihydrotestosterone complex associated with the chromatin was found to be preferably distributed into the non-histone protein as well as the DNA itself of the chromatin. The radioactivity lost by dehistonization of the chromatin was almost negligible. Furthermore, the cytosol 9S receptor fraction bound to ³H-dihydrotestosterone was purified about 100 fold by the two consecutive column chromatographies. The partially purified receptor fraction which was labeled with radioiodine incorporated mostly into nonhistone protein and DNA fractions.

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Effect of implantation of estrogens or catecholamines into the medial preoptic area through chronically implanted double cannula on the release of LH was examined in the ovariectomized estradiol-primed rat. Implantation of estrone at 12:00 on the second or third day after subcutaneous injection of estradiol benzoate (20μg) increased the serum concentration of LH at 18:00 compared with that in the nonimplanted controls, whereas implantation of estradiol benzoate on the third day did not affect it. Dopamine implantation at 12:00 on the third day also induced a significant increase of LH concentration at 18:00, and, in contrast, norepinephrine implantation decreased the concentration at 18:00.
It may be said that the medial preoptic area is responsive to estrone and can induce LH release, whereas it is not to estradiol. Furthermore, dopamine was effective for the activation of the medial preoptic area in relation to the inducement of LH release.

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A case of Cushing's syndrome due to huge adrenocortial multinodular hyperplasia who was shown to be hyperresponsive to ACTH administration, unresponsive to metyrapone administration and resistant to dexamethasone high dose suppression was reported.
After two years' duration of his symptoms, the multinodular adrenals weighing 161 g in total were removed by bilateral adrenalectomy which abolished his symptoms. Postoperatively, plasma ACTH rose gradually to above normal levels, suggesting the presence of primary disorder in the hypothalamus-pituitary axis.

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In Experiment 1, female rats were given a single subcutaneous injection of 1.25mg 5α-dihydrotestosterone (DHT) or its propionate (DHTP) on day 5 of postnatal life. All of them showed regular estrous cycles as adults like untreated control animals. At about 60 days of age, the rats were ovariectomized and given 7 daily injections of 2mg progesterone (P) plus 0.2μg estradio1-17β(ED). Uterine trauma applied on the 4th day of P-ED injections resulted in well developed deciduomata in all animals by the day after the last injection. This made a sharp contrast to the failure of female rats receiving testosterone propionate (TP) neonatally to give a positive response under similar experimental conditions (Takewaki and Ohta, 1974). The mean weight of traumatized horns was significantly larger in DHTP-treated rats (but not in DHT-treated rats) than in controls.
In Experiment 2, rats were ovariectomized on day 4 and given a dose of 1.25mg DHT or DHTP on day 5. Controis were ovariectomized on day 4 but not given any steroid on the next day. A series of 7 daily injections of 2mg P plus 0.2μg ED was started at about 60 days of age, after the animals had received 3 daily injections of 0.2μg ED or 30 daily injections of 0.1μg ED. Incidence of deciduomata following uterine traumatization was markedly lowered only in animals treated with DHTP neonatally and given 0.1μg ED for 30 days as adults, no significant differences being found in both incidence and size of deciduomata among the other groups.
It was suggested that the effects of neonatal steroid administration on uterine responsiveness in adulthood are specific to the steroid. The previous conclusion that persistent estrus in androgen-sterilized rats plays a part in the reduction of uterine responsiveness was confirmed. An exposure of rats to estrogen for a prolonged postpuberal period was without effect, unless the animals had received enough androgen neonatally.

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In the smallest biologically active fragment of angiotensin or a 3-8 hexapeptide, the C-terminal was substituted by isoleucine residue. It proved to be the smallest potent antagonist of angiotensin II so far reported.

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Insulin concentration in the peripheral blood and the thoracic duct lymph from male rabbits with CCl4-induced liver cirrhosis was measured using a radioimmunoassay technique. Although insulin concentration in the lymph was lower in the cirrhotic animals than in the control ones, the hourly transport of the hormone by the lymphatic vessel of the cirrhotic animals markedly increased in company with the higher flow rate of the lymph.

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Effects of monoamines or its precursors on the induction of persistent estrus by testosterone propionate (T.P.) were studied in Wistar strain female rats.
Daily treatment with 5-hydroxytryptophane (5-HTP) for the first 10days of life delayed the occurrence of persistent estrus in rats given T.P. at 4 days of age. The onset of persistent estrus occurred in this group of animals at 67.1±2.5days of age compared with 45.1±1.4days of age for the saline treated control group. The role of hypothalamic monoamines in contributing to the induction of persistent estrus by T.P. treatment of neonatal rats is discussed.

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One mg of glucagon was given subcutaneously to eight patients with active acromegaly. Seven out of eight patients had a rapid decrease in serum growth hormone (GH) levels at 30 min after the glucagon injection. In two out of seven patients a rebound increase in serum GH following the early GH reduction was observed.
On the other hand, oral administration of 50g glucose which caused a comparable increase in blood glucose to that after the glucagon injection elicited no early suppression in serum GH levels in the same patients. These data suggest that the inhibition of GH release induced by glucagon could not be related to the increase in blood glucose by glucagon.

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The biological properties of human chorionic FSH (hCFSH) for rat ovaries were investigated. Highly purified hCFSH had similar response to the ovarian augmentation test as bovine FSH and significantly enhanced ³H-thymidine uptake by granulosa cells and theca cells in the ovary of hypophysectomized rat. In contrast, highly purified hCG little responded to the ovarian augmentation test and had no effect on ³H-thymidine uptake by the ovary. These results indicate that hCFSH may promote the follicular growth of ovary resulting from granulosa cell proliferation and its enlargement. In addition, freshly harvested porcine granulosa cells were employed in an in vitro system to investigate specific binding of hCFSH to ovarian receptor. Radioiodinated hCFSH (¹²⁵I-hCFSH) and hCG (¹²⁵I-hCG) were respectively incubated with cell suspensions. Binding of these hormone preparations was proportional to the cell number and increased with the time of incubation through 120 minutes. The binding ability of ¹²⁵I-hCFSH to the cells was greater than that of ¹²⁵I-hCG. Increasing concentrations of unlabeled hCFSH in the incubation mixture progressively inhibited the uptake of ¹²⁵I-hCFSH by granulosa cells. Unlabeled hCG was not able to compete with ¹²⁵I-hCFSH binding. The similar, phenomenon to inhibit the binding of ¹²⁵I-hCG to the cells was also recognized in the presence of unlabeled hCG. These findings suggest that granulosa cell has at least two different types of receptor sites: one for hCFSH and the other for hCG.

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