This study was designed to investigate the oxytocin (OT) specific binding receptors in 20, 000×g pellets of nonpregnant, first trimester and term human myometria. The receptor analysis was done using the lower uterine segment at term and the lower portion of the anterior uterine body in nonpregnant and first trimester subjects, and no difference was found in the myometrial receptor concentrations in the various uteri. The mean±S.D. values of the receptor dissociation constants were 3.33±0.50, 2.71±1.03 and 1.87±0.30nM and the number of binding sites was 0.30±0.10, 0.50±0.10 and 1.50±0.50pmol/mg protein at each stage studied, indicating that the gestational increase of uterine sensitivity to OT is due to the increase in myometrial OT binding sites as well as its binding affinity. Further, myometrial OT binding before and after the onset of labor was studied and a marked decrease in total myometrial OT binding was noticed; 35.6±13.0% before and 20.2±5.0% after. This decrease was thought to be due to the decrease in the number of binding sites from 1.50±0.50 to 0.74±0.21pmol/mg protein after the onset of labor (p<0.01). No changes Were found in the dissociation constants. Thus it seems that OT and its receptor coupling triggers labor or is involved in the early steps of labor.

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A morphological and biochemical study was performed on thyroid tissue with various thyroid diseases. The thyroid peroxidase (TPO) activity of normal thyroid tissues ranged from 2.6 to 7.0m GU/mg DNA. The activity was low in adenomas and extremely low in carcinomas, and there was no significant relationship between the histological subclassification of follicular adenomas (simple, colloid, oxyphil) and TPO activity. The activity was various in the cases of chronic thyroiditis, ranging from non-detectable to 9.8m GU/mg DNA, and the TPO activity showed a close correlation with the degree of lymphoid cell infiltration of the diseases. In the seven cases of Graves' disease, the values were high, though the elevation was not so remarkable in three cases which had already been euthyroid or slightly hypothyroid after long-term treatment.
By means of subcellular fractionation, more than 50% of peroxidase activity was shown to be localized in the microsomal pellets, and this result well coincided with the electron microscopic findings of prominent development of rER.

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Human decidua, chorion, amnion and placenta from the 1st., 2nd. and 3rd. trimesters of gestation were investigated for synthesis and secretion of prolactin by in vitro incubation of these tissue fragments in medium 199. Prolactin content in deciduawas found to be significantly higher than that in chorion, amnion or placenta at any stages of gestation. During 6hours of incubation, decidua secreted significantly more prolactin into medium than did chorion, amnion or placenta. The amount of prolactin secreted by decidua was significantly higher than the prolactin content in tissue before incubaton. Decidua were also incubated in medium 199 with or without actinomycin-D (20-200μg/ml), puromycin (200μ/ml) or cycloheximide (100-200μg/ml) for 12hours. Both total prolactin secreted into medium and prolactin content in tissue after incubation were significantly lower than the control without inhibitor. Decidua of 2nd. trimester of gestation was noted to secrete more prolactin into medium than decidua of 1st. or 3rd. trimester of gestation. In further studies, ³H-leucine (200μCi) was incubated for 12hours with decidua (10 grams) from 2nd.trimester of gestation. The incorporated protein in medium or tissue was extracted, fractionated by gel filtration through Sephadex G-100 column and by 10% SDSpolyacrylamide gel electrophoresis. Peaks of 3H count and immunoreactive prolactin in gel slices were coincident with the peak of standard pituitary prolactin at gel slices' No.31 (Rf=0.62). These results demonstrate that prolactin is synthesized and secreted by human decidua, which is identical to human pituitary prolactin. The synthetic activity is most prominent in the decidua of the 2nd. trimester of pregnancy.

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The effect of calcitonin on the metabolism of calcium and cyclic AMP in the avian bone was examined in vitro. In the chick embryonic cortical bone, calcitonin affected neither the level of cyclic AMP nor the parathyroid hormone-induced stimulation of calcium release from bone. Also, the level of cyclic AMP in the cortical or medullary bone of Japanese quails was not increased by calcitonin. The medullary bone did not respond to calcitonin either in freshly prepared tissue or in tissue cultured for 48hr in the absence of calcitonin. The activity of renal adenylate cyclase of Japanese quails was also calcitonin-insensitive. The inability of calcitonin to increase the level of cyclic AMP and to antagonize the parathyroid hormone-induced stimulation of calcium release from bone may account for the lack of hypocalcemic effect of calcitonin in birds as reported by other investigators.

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The lipid materials extracted from hog thyroid microsomes were found to contain factors which stimulated both the NADPH oxidation by NADPH-cytochrome c reductase and PBI formation by a reconstituted system containing the reductase as a hydrogen peroxide generator and thyroid peroxidase. For both reactions, the stimulating activity in the extracts from the other subcellular fractions was less than that observed in the extracts from microsomal fraction. Lipids extracted from hog liver microsomes did not show any NADPH oxidation stimulating activity.
Thin-layer chromatography of the lipid extracts from thyroid microsomes revealed the presence of at least two stimulating factors differing in chromatographic behaviour. At least one of the stimulating factors separated on TLC stimulated the oxygen consumption accompanied by generation of hydrogen peroxide due to the NADPH oxidation by NADPH-cytochrome c reductase.
From these results it was suggested that in thyroid microsomes, but not in liver microsomes, there were stimulating factors extractable by chloroform-methanol. The factors could stimulate NADPH oxidation by NADPH-cytochrome c reductase followed by generation of hydrogen peroxide, resulting in the enhancement of PBI formation coupled with thyroid peroxidase.

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Antisera having a strong and strictly constant cross-reactivity for gut-GLI were raised in seven rabbits immunized with immunogen, a conjugate of BSA and des-Asn²⁸, Thr²⁹, Homoser²⁷-glucagon (CNBr-glucagon). All of the anti-CNBr-glucagon sera exhibited titer and affinity for glucagon sufficiently high enough to develop a sensitive radioimmunoassay. The relative crossreactivity of the antisera to gut-GLI was comparable to that of antiserum K-4023 which strongly crossreacted with gut-GLI. One of the anti-CNBr-glucagon sera, OAL-196, did not react with the glucagon 19-29 fragment at all. The intra-and inter-assay coefficients of variation were 3.8-5.0 and 6.0-7.3%, respectively, in the radioimmunoassay system for total GLI in human plasma using OAL-196. The fasting plasma total GLI was 374±18pg/ml. The plasma total GLI during 50g oral glucose load in normal subjects increased significantly, whereas the plasma IRG level measured with the anti-glucagon 19-29 serum, OAL-123, assay system was lowered. In the gastrectomized subjects, plasma total GLI measured with the present assay system elicited a marked increase following an oral glucose load. These results suggest that the radioimmunoassay using anti-CNBrglucagon sera will be useful in measuring plasma total GLI.

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The presence of complement-fixing islet-cell antibodies (CF-ICA) and islet-cell antibodies (ICA) was examined in 355 patients with different types of diabetes mellitus in the Japanese population by an indirect immunofluorescence test (IFT).
The overall prevalence of ICA, which were stained as a homogenous cytoplasmic fluorescence in islet cells, was 7 per cent (5/67) in insulin-dependent (Type I) diabetics, 4 per cent (6/137) in noninsulin-dependent (Type II) diabetics treated with insulin and 2 per cent (1/58) in Type II diabetics treated with oral hypoglycemic agents. None of 84 Type II diabetics receiving diet alone and 9 diabetics associated with chronic pancreatitis had ICA. CF-ICA, which were stained as a “ring-shaped” fluorescence in a part of the cytoplasma, were demonstrated in 5 out of 12 cases (42%) whose sera possessed ICA.
The lower prevalence and remarkably shorter persistence of ICA and CF-ICA in Japanese diabetics than those observed in Caucasian diabetics may be explained by the heterogenous immunological response in different races or possible heterogeneityof Type I diabetics.

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In an attempt to clarify the underlying mechanism (s) in the disappearance of phosphaturic response to bolus parathyroid hormone (PTH) in hyperparathyroid patients, the effects of bolus bovine PTH (10 USP U) were studied in conscious thyroparathyroidectomized (T·EPTX) male Wistar rats that had been infused with a dose of PTH (2.5U/hr, for 16 hours) so as to reproduce hyperparathyroidism. These animals responded with an increase in urinary cyclic AMP, but without an increase in renal clearance of phosphate.
The loss of phosphaturic response was not prevented by pretreatment with ac-tinomycin D at a dosage close to full toxicity (0.1mg/kg BW, ip, for 3days). Ac-tinomycin D at this dosage did not affect the normal stimulatory effects of bolus PTH on urinary cyclic AMP and renal clearance of phosphate in T·PTX rats. The con-tinuous infusion of PTH produced nearly maximal phosphaturia throughout in the face of a significant depletion of phosphate. In addition, pretreatment with actino-mycin D did not cause a further increase in urinary phosphate excretion during the infusion.
These results, along with the report of Shah et al.(1979) indicating that the de-velopment of antiphosphaturic adaptation to acute phosphate depletion was prevented by comparable amounts of actinomycin D, indicate that the disappearance of phos-phaturic response to bolus PTH by prior PTH infusion simply signifies the continua-tion of maximal phosphaturic response to the preceding PTH infusion. It is also suggested that the continuous action of PTH prevents, at least phenomenologically, the development of the gene-activation-mediated refractoriness to PTH or antiphos-phaturia induced by acute phosphate depletion.

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The effects of intrahepatic portal infusion of hyperosmotic substances or angiotensin II (A II) on the plasma arginine vasopressin (pAVP) concentration were investigated. At the beginning of each experiment, rats anesthetized with sodium pentobarbital were infused with 0.9% NaCl at a rate of 0.037ml/min via the hepatic portal vein for 30min. Subsequent intraportal infusions of 1.4, 1.8 and 2.7% NaCl for 5min at a rate of 0.052ml/min raised pAVP levels from 15.6±5.2pg/ml to 26.0±1.7, 61.5±8.8 and 37.5±6.6pg/ml, respectively. However, subsequent intraportal infusions of 0.9 or 3.6% NaCl and intrajugular infusions of 1.8 or 3.6% NaCl did not further raise the pAVP levels. Intraportal infusions of 21 and 42% sucrose raised pAVP levels to 28.3±2.9 and 24±2.0pg/ml, respectively, whereas glucose did not changethe pAVP concentration.
When A II was infused via the intraportal vessel at a rate of 30ng/kg/min for 10, 15 or 30min, pAVP levels became 29.3±2.7, 32.1±4.0 or 25.8±3.2pg/kg/min respectively. PAVP levels did not change when the same dose of A II was infused into the jugular vein. The increase of pAVP induced by A II was prevented when angiotensin II analogue (A II A), ¹Sar, ⁸Ileu-A II, was infused into portal vein simultaneously.
These results not only suggest that the osmoreceptors exist in the area locatedsomewhere within the hepatic portal vascular bed, but also that the osmoreceptors are somewhat different in nature from those of hypothalamic osmoreceptors. Furthermore, these results su

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A possible metabolic linkage between hepatic thyroxine (T4) 5'-deiodination and the NADPH-glutathione (GSH) cycle was studied in rat liver. Supplementation of 1mM NADPH to stocked liver homogenates in vitro produced 4 fold increase in 3, 5, 3'-triiodothyronine (T3) formation from T4, whereas the effect of 1mM FMN, FDN, NAD, NADH, or GSH was relatively small. An exponential dose-response relation was obtained between NADPH and T3 generated. The dose-dependent increase in T3 formation on GSH was eliminated in the presence of 1mM MADPH, and the additive effect of GSH to NADPH was not apparent in comparison with NADPH alone. Inhibition of T3 generation by graded doses of methylene blue was not affected by the presence of 5mM GSH. Furthermore, metabolic changes in the hexose-monophosphate shunt were produced in male Wistar rats aged 5w by treating them with fastingrefeeding (FF group), with the administration of insulin. and glucose (IG group), with propylthiouracil (PTU group) and with T4 (T4 group). All these reatments significantly reduced hepatic T4 5'-deiodinase activity (P<0.01-0.001 vs control), while glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GSSG-R) activities were increased. Between generated T3 and G6PD or GSSG-R activity, an inverse correlation was noted (r=-0.802 and -0.933, P<0.001). No consistent relation was found between T4 5'-deiodinase activity and GSH or non-protein SH contents. The addition of 1mM NADPH and GSH to the homogenates of FF, T4 and the control group stocked for 4w at -20°C, restored T4 5'-deiodinase activity from a level of 10% to 60% of the initial value, whereas the activity remained depressed in PTU (19%) and the IG group (37%). These results indicate that both GSH and NADPH are important cofactors of the T3 generating system, but NADPH is more rate-limiting and its effect appears to be rather direct, not mediated by GSH formation. It is possible that T4 5'-deiodinase may be one of the NADPH-dependent enzymes.

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Thyroxine (T₄) 5'-monodeiodinating activity in rat pituitary was studied, using paper chromatography. Rat anterior pituitary homogenates were incubated with ¹²⁵I-T₄ , dithiothreitol and unlabeled L-T₄ at 37°C. The incubation mixture was extracted by ethanol and the extracts were subjected to descending paper chromatography. The conversion ratio of T₄ to 3, 5, 3'L-triodothyronine (T₃) was calculated from the radioactivity of T₃spot to total radioactivity on the paper strip. The T₃amount generated was estimated from the T₄ concentration in the incubation mixture and the conversion ratio.
The T₃generating activity from T₄ increased as the amount of tissue increased. It was temperature-and pH-dependent, and thiol sensitive. These results suggest the enzymatic nature of T₄ 5'-monodeiodinating activity of the anterior pituitary. A kinetic study revealed low Km for T₄ (7.9±1.6nM, Mean±SE), with Vmax of 68.0±12.7 fmoles T₃/mg Protein/min.
T₄ monodeiodinating activity was consistently, though minimally, detected in fetal rat pituitaries and increased after birth, reaching the maximum at 22days. It declined thereafter to the young adult level. Serum TSH levels were markedly elevated in fetus. They decreased after birth, reaching the nadir at 22days, and then increased to young adult levels. Serum T₄ and T₃levels were markedly diminished in fetus and gradually increased after birth, reaching the young adult levels at 17 days. Thus, a reciprocal relationship was observed between intrapituitary T₃generating activity from T₄ and serum TSH levels in developing rats. It suggests that the conversion of T₄ to T₃in the pituitary plays a role in regulating TSH secretion.

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The substrate specificity of rat testicular and epididymal peptidase was investigated using chromogenic substrates, D. L-alanine-, L-arginine-, γ-N-L-glutamine-, L-leucine-, D. L-methionine-, α-N-benzoyl-D. L-arginine-, and N-benzoyl-L-leucine-β-naphthylamide. The histochemical distribution of peptidase activity demonstrated with these substrates was also investigated in the testis and epididymis.
L-Arginine-β-naphthylamide (Arg-β-NA) and γ-N-L-glutamine-β-naphthylamide (Glu-β-NA) were mostly hydrolyzed in the testis and epididymis, respectively.
Histologically, the activity using Arg-β-NA as substrate (animopeptidase B) appeared in both the cytoplasms and nuclei of interstitial cells and spermatogonia and the heads of spermatozoa, while activity using other substrates was found only in the cytoplasms of cells in the germinal epithelium. In the epididymis, strong reaction with Glu-β-NA (γ-glutamyl transpepetidase) was found in the apical part of the epithelial cells and in the heads of spermatozoa.
Neither α-N-benzoyl-L-arginine-nor N-benzoyl-L-leucine-β-naphthylamide was utilized in either the testis or the epididymis.

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Stereological analysis at the electron microscope level was performed on zona fasciculata and zona reticularis of the adrenal cortex of male rats maintained on normal rat chow, clofibrate (hypolipidemic drug), cholesterol-rich diet or cholesterolrich diet plus clofibrate. The purpose of this study is to know whether the alteration in the serum cholesterol level is accompanied by an alteration in the ultrastructure and steroidogenic activity of the adrenal cortex, and to compare these changes with those of Leydig cells reported previously.
The corticosterone level in both serum and the adrenal tissue of cholesterol-fed rats remained unchanged. Treatment of the cholesterol-fed rats with clofibrate caused a significant rise in the serum corticosterone level but the concentration in the adrenal remained unchanged.
Lipid droplets increased in number and volume significantly in both cortical zones in all experimental groups as compared with the control group. But neither cholesterol feeding nor clofibrate treatment of normal rats caused a change in the volume or surface area of smooth endoplasmic reticulum (ER). Treatment with clofibrate on the cholesterol-fed rats resulted in a decrease in the volume of smooth ER in both cortical zones. Mitochondria showed no change except for a decrease in volume and inner membrane surface area of zona reticularis of cholesterol-fed rats. Peroxisomes did not proliferate but rather decreased in clofibrate-treated rats.
These changes in the adrenal cortex were different from those in Leydig cells. Possible reasons for these differences were discussed.

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Insulin responses during 100g glucose tolerance tests (GTT) were compared between three groups of patients with varying degrees of glucose intolerance. Patients who had no disease known to be associated with secondary diabetes were classified as patients with idiopathic diabetes mellitus. Those whose present and past fasting blood glucose (FBG) exceeded 140mg/100ml were assigned to Group A, and the rest of the patients to Group B. Group C included patients with liver disease, thyrotoxicosis, or myocardial infarction, or those treated with corticosteroids or who had undergone gastrectomy. Patients in Group A were found to have consistently subnormal insulin responses whether glucose tolerance was normal (i.e. previous abnormality of glucose tolerance), borderline, or diabetic. In contrast, patients in Group C without fasting hyperglycemia had enhanced rather than decreased insulin responses when glucose tolerance was the more impaired. Patients in Group B had insulin responses similar to those either of Group A or of Group C.
The relationship between the sum of six insulin and six blood glucose values during GTT (ΣIRI and ΣBG) was examined. The ΣBG-ΣIRI plot revealed distinctly different distribution zones for Group A and Group C (Zones A and C). In Group A, ΣIRI values were below 300μU/ml irrespective of ΣBG values. In Group C, ΣIRI tended to increase, paralleling the increase in ΣBG values in the range of ΣBG values lower than 1400mg/100ml. In patients whose ΣBG rose above 1400/100ml during corticosteroid treatment, the ΣIRI values decreased and entered into Zone A. After the cessation of corticosteroids in a few of these patients, the ΣIRI values recovered and reentered Zone C, concomitant with an improvement in glucose tolerance. Similar recovery of insulin response from Zone A to Zone C was also observed after the treatment of two obese diabetic patients. Thus, patients with glucose intolerance due to extra-pancreatic causes may secrete insulin at a higher rate than normal so long as the FBG level remains below 120mg/100ml, but a further deterioration in glucose metabolism may lead to a failure of insulin secretory mechanisms.

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When [4-14C]-5α-dihydrotestosterone was incubated with the homogenate of human epididymis, 5α-androstane-3α, 17β-diol and 5α-androstane-3β, 17β-diol were identified as major metabolites. The ratio of 3α- to 3β-epimer in androstanediol formation was approximately 2.4. 5α-Androstane-3, 17-dione was also identified as a minor metabolite. Among the subcellular fractions, both the human epididymal 3α-and 3β-hydroxysteroid dehydrogenases were localized almost exclusively in the cytosol fraction (105, 000×g supernatant). Both enzymes had optimum pH at 7.5 and optimum temperature at 46°C. NADPH was a more preferable cofactor than NADH for both dehydrogenases. The Michaelis constants (Km) of 3α- and 3β-hydroxysteroid dehydrogenase for 5α-dihydrotestosterone were similar and estimated as 8×10-5M, but the enzymes were unsaturable with the substrate under the conditions investigated, indicating low affinity and high capacity of both dehydrogenases for 5α-dihydrotestosterone.
The human epididymal 5α-reductase revealed a regional difference in activity. The 5α-reductase activity in the most proximal part of the head (ductuli efferentes) was one seventh to one tenth the activity in the remaining part of the epididymis which was constructed of ductus epididymis. Except for this finding, the activity of 5α-reductase was highest in the head, then declined along the course to the tail portion. The 5α-reductase for testosterone was competitively inhibited by Δ4-3-oxosteroids such as progesterone, 20α-dihydroprogesterone, 17α-hydroxyprogesterone, 4-androstenedione, 11-deoxycorticosterone, corticosterone and 11-deoxycortisol, which had inhibition constants (Ki) of 3.3×10-^-9M, 2.2×10^-9M, 1.8×10^-8M, 1.3×10^-8M, 8.3×10^-9M, 1.5×10^-7M and 8.7×10^-8M, respectively, suggesting the possibility that the 5α-reduction of testosterone is regulated by other Δ4-3-oxosteroids.

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In order to evaluate the effect of sex and age on the protrusion of the eyes in patients with Graves' disease, exophalmometry was performed on 95 patients, aged 16-73. The mean (±SE) protrusion in the 190 eyes was 17.5±0.2mm, being significantly higher than those in normal subjects (14.9±0.2mm) and patients with Hashimoto's disease (14.3±0.2mm). No significant difference existed in the degree of protrusion between thyrotoxic Graves' disease patients and those maintained in an euthyroid state during treatment with antithyroid drugs. When compared in terms of the duration of the treatment, the protrusion became prominent with the increased duration of the treatment, especially in male patients, although statistically not significant. The protrusion in male patients tended to increase, though not significantly, with increasing age, whereas it decreased significantly with increasing age in female patients.

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The effect of testosterone propionate (TP) on the recovery of spermatogenesis was compared in 3 types of experimentally induced gonadotropin-deficient rats; 1. hypophysectomized rats, 2. rats actively immunized to LHRH and 3. hypophysectomized and immunized rats bearing a pituitary isograft. In order to immunize animals to LHRH, deamidated LHRH conjugated with BSA (LHRH-BSA) was injected intradermaly to male rats 4times at 2week intervals and additional booster injections continued once a month.
Anti-LHRH titer was the highest on the 12th week and the titer remained at a relatively high levels thereafter. A remarkable decrease in testicular weight and testosterone production was achieved in 10weeks associated with a drop in serum LH and FSH levels. In long term immunized rat (LIM rats) which had been immunized for more than 3months, testicular atrophy advanced to a similar extent to that in long term hypophysectomized rats (HX rats).
The administration of TP (1mg/day) sc for 30days restored spermatogenesis in LIM rats. The simultaneous administration of anti-LH and-FSH sera in addition to TP did not affect the restoration of spermatogenesis in LIM rats. The same TP treatment in HX rats, however, failed to restore spermatogenesis. Spermatogenesis was reinitiated after TP treatment if immunized-hypophysectomized rats received a pituitary isograft. Though the serum PRL level in LIM rats was one fifth of that in normal rats, a 2.5-fold rise in the PRL level was observed after TP treatment.
These results suggest that PRL is involved in the process of spermatogenesis.

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Female rats given neural transection between suprachiasmatic and arcuate nuclei at 2days of age showed regular sexual cyclicity and had ovarian corpora lutea. Thus, the ovarian activity seemed identical to that of intact or sham-operated control rats. However, the diurnal rhythm of the blood corticosterone concentration of the neonatally transected rats was not evident at 22days of age, while it was already evident in controls. At 30 and 50days of age, they had clear adrenal rhythms although the peak values were significantly lower than those of controls. Thus, the anterior connections of the medial basal hypothalamus, which are necessary for both cyclic gonadotropin discharge and diurnal corticosterone secretion, are still plastic and not yet established at 2days of age.

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5-HTP (5mg/100g body weight), a precursor of serotonin, was administered intravenously to pituitary stalk sectioned rats. Plasma prolactin levels were initially increased 30 and 60min after the injection of 5-HTP and thereafter decreased. Pituitary prolactin content was decreased 60min aftr the stalk section as compared to those seen following sham operation. The treatment of 5-HTP induced a moderate increase in pituitary prolactin content in the stalk sectioned rats.
In order to confirm the complete disconnection of pituitary stalk after the operation, 2.5μg of dopamine hydrochloride was given into the lateral ventricle of stalk sectioned rats. Plasma prolactin did not change following the administration.
Our results demonstrate that 5-HTP acts either directly at the pituitary level or via some mechanism yet to be determined.

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To investigate localization of the action on blood pressure in the peptide chain of parathyroid hormone (PTH), we determined the effect of seven fragments of human (h) PTH-(1-34) fragments on blood pressure in anesthetized rats. hPTH-(1-7) and-(28-34) fragments had no effect up to 1×10^-6mole/kg. hPTH-(8-34), -(13-34), -(18-34), and-(23-34) fragments showed significant hypotensive activity. The decrease in the chain length lowered the hypotensive activity. The results indicate that some amino acids in position 23-34 are necessary for the hypotensive activity. This accords with the previous report suggesting that 5 amino acids in position 24-28 are essential. In addition hPTH-(8-34) and-(13-34) fragments showed initial transient hypertensive activities. hPTH-(1-13) fragment had only hypertensive action, suggesting that some amino acids in position 8-13 are necessary for hypertensive activity.

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