To investigate the intestinal absorption of parotin, ¹²⁵I-labelled parotin was added in the mucosal side of the sac of rat everted intestine and incubated for 1 hr. Parotin seemed to be transmitted through the intestinal wall into the serosal fluid, because the non-diffusible ¹²⁵I-bound substance which reacted with the anti-parotin rabbit serum was detected in the serosal fluid. The amount of ¹²⁵I-macromolecule in the serosal fluid which was estimated from the radioactivity was in good agreement with that of the immunoreactive substance in the fluid determined by a single radial immunodiffusion, and the concentrations of these substances in the serosal fluid were about 1/5 of parotin concentration in the mucosal fluid.
Apart from this, in situ mesenteric perfusion was performed on the rats which were administered parotin and ¹²⁵I-labelled parotin in the ligated jejunum. The perfusate was collected from the mesenteric vein for 2 hr, and ¹²⁵I-bound macromolecule which corresponded to 3.4% of the administered ¹²⁵I-labelled parotin was found in the perfusate. There was an immunoreactive substance with the anti-parotin serum in the perfusate, too.
From these results, it was inferred that about 3% of parotin given into the intestine was transmitted into the mesenteric blood vessels.

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A diabetes-like state was induced by intravenous injection of a single dose of streptozotocin (SZ), 45mg/kg, in pregnant rats and the cholesterol level in the tissues of. the mothers and their infnats was examined during the early postpartum period.
The following results were obtained.
1) The infants of the diabetic mothers were smaller than the infants of normal mothers and were markedly hyperglycemic.
2) Serum cholesterol was markedly increased on day 21 of pregnancy both in normal and diabetic groups. The concentration, however, declined nearly to the nonpregnant normal level on the 1st day after delivery in normal mothers, while it decreased stepwise until day 7 in diabetic mothers.
3) The cholesterol contents in the adrenal, liver and serum of infants of normal and diabetic groups increased progressively during the early postpartum period. However, the cholesterol level in these tissues was always lower in the infants of diabetic mothers than in normal infants; the concentration in the liver was considerably lower.
4) The adrenal cholesterol concentration decreased abruptly in the normal mother on day 1 after delivery and it returned to approximately normal level on day 7. On the other hand, no alteration in the cholesterol concentration was observed in the daibetic mother after parturition. In addition, the adrenal weight of the diabetic mother was heavier than that of the normal control mother at all stages examined.
These results suggest impairment of the steroidogenesis in the maternal adrenal in response to endogenous ACTH during the parturition in a diabetic state. Moreover, it was clearly demonstrated that SZ-induced hyperglycemia in pregnant rats affected the cholesterol metabolism in the adrenal and liver in newborns.

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Salmon calcitonin was administered intravenously at a dose of 5 MRC units per day to a patient with prostatic cancer and bone metastases during 20 days period. Serum calcium maintained pretreatment level during the period of administration with a slight decrease after cessation of the treatment. Urinary total hydroxyproline showed a gradual decrease during the treatment. Serum alkaline phosphatase increased gradually, and the elevated level continued even after cessation of the administration. Urinary phosphorus increased transiently after the initiation of the administration.
When hexestrol was administered intramuscularly at a total dose of 765 mg, enlarged cervical lymph nodes and metastatic lung infiltration disappeared completely at the end of the treatment. Urinary hydroxyproline began to increase at the initiation of administration, and the high value was maintained during and after the hexestrol therapy.

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A radioimmunoassay for serum dehydroepiandrosterone has been developed by using the anti-DHEA antiserum. After DHEA-7α-³H was added to 0.2 to 1.0ml of serum and 1ml of water for recovery, DHEA was extracted with methylene chloride, and the extract was evaporated to dryness. The residue was applied to paper chromatography. The eluates and DHEA-7α-³H which was added to determine the % free of DHEA were evaporated to dryness, and the residues were incubated with the antiserum containing pepsin treated human immune serum globulin and bovine serum albumin at 4°C overnight. Ammonium sulfate was used to separate free from bound DHEA. The accuracy, precision and specificity were satisfactory. The sensitivity was 300pg per sample. The blank value could not be differentiated from zero. Although the antiserum reacted with the other 3β-OH-Δ⁵-steroids as well as with DHEA, complete separation of DHEA from the other 3β-OH-Δ⁵-steroids was achieved chromatographically. Serum DHEA levels in normal subjects and their responses to ACTH stimulation and dexamethasone suppression were evaluated by using the radioimmunoassay.

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Effect of neonatal treatment with synthetic LH-RH on the development of gonadotropin (GTH) regulation was studied. Newborn male rats were injected with 0.01-1μg LH-RH from Day 1 (=the day of birth) to Day 2, and castrated on Day 3. Significant stimulation of testicular weight and histology was recognized in the rats receiving 0.1 or 1μg of LH-RH when examined upon castration on Day 3. To estimate the secretory pattern of GTH ovarian transplantation was performed around Day 30. At autopsy on Day 60, 100% of rats receiving 1μg LH-RH were masculinized. All the rats had polyfollicular masculine ovarian grafts. When the dosage decreased to 0.1μg, 60% of the injected rats were masculinized. A further decrease in the incidence of masculinized males were obtained by injection of 0.01μg of LH-RH: 40% of the treated rats were of the masculine type. In these males, advancement of differentiation of the masculine pattern of GTH secretion apparently occurred, because neonatal castration on Day 3 failed to produce “feminine males.” In contrast, rats treated with saline alone became “feminine males.” Most of this group of rats had a typical feminine ovarian graft which contained numerous healthy corpora lutea and developing follicles. These results suggest possible participation of the hypothalamic-pituitary axis in the masculinizing process of GTH regulation.

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Role of the bed nucleus of stria terminalis (BST) in the control of gonadotropin release in the female rat was studied.
Electrical stimulation of the BST on the day of diestrus II increased the serum concentration of FSH, but not of LH. The stimulation on the day of estrus increased serum concentration of LH as well as on the day of proestrus. In the ovariectomized and estrogen-treated rat, the stimulation decreased serum concentration of FSH; but in the ovariectomized and progesterone-treated rat the stimulation did not decrease it. Implantation of estrogen into the BST inhibited th castration-induced increase of LH and FSH in serum.
These results indicate that the BST is one of negative feedback sites of estrogen, and probably participates in folliculotropin secretion which is responsible for estrogen secretion before proestrus. The physiological significance of its ability to induce LH release on the day of estrus in response to stimulation is not known now.

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In order to evaluate the effect of treatment for hyperthyroidism based on a proposed criteria from our institution, the patients with the disease first seen at the Ito Hosiptal during 1965 were studied on the basis of clinical diagnosis and the follow-up study in 4-5 year period.
Surgery was recommeded in the younger ages and radioiodotherapy in the higher ages for both sexes, though there was no inclination for antithyroid therapy in ages. Surgery exihibited the most reliable result among the three major therapies. Hypothyroidism was found more often in females than in males. Incidence of hypothyroidism with radioiodotherapy after the first four years of therapy was about 10% in females, and the result indicated the necessity of further improvement for administering ¹³¹I. In case of radioiodotherapy, it is difficult to predict the manifestation of hypothyroidism, because of no relationship between the incidence of hypothyroidism and radiation dose. The follow-up study showed the remission rates with surgery and radioiodotherapy were both 80-90 per cent, whereas the rate with antithyroid therapy was 38 per cent.

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Radioimmunoassay of human C-peptide was established using antiserum to synthetic human connecting peptide and synthetic tyrosylated human connecting peptide. Synthetic human connecting peptide and natural human C-peptide showed the same degree of cross-reactivity with the synthetic human connecting peptide antiserum. Natural human proinsulin reacted approximately one fourth as well as the human connecting peptide or natural human C-peptide when expressed on an equimolar basis.
Synthetic porcine and bovine connecting peptide, human insulin and natural porcine proinsulin did not react with the human connecting peptide antiserum. Synthetic human connecting peptide did not cross with insulin antiserum. The senstitivity of the radioimmunoassay was 0.05ng/tube.
The assay system was available to determine C-peptide immunoreactivity in plasma, because proinsulin and proinsulin-like components had been reported to be very low in plasma. The fasting level of C-peptide immunoreactivity in plasma of normal subjects assayed by this system was 0.88±0.21ng/ml.

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The secretion of endogenous insulin was investigated through the effect of the infused exogenous bonito insulin on dogs. The systemic infusion of small doses of exogenous bonito insulin (0.3mu/kg) which proved inert on the blood sugar levels, resulted in an inhibition of endogenous insulin secretion. The prolonged infusion of exogenous bonito insulin inhibited both the basal insulin secretion and the biphasic pattern in glucose-induced insulin release. The mean level of endogenous insulin in the pancreatic vein decreased significantly (p<0.05) from 453±97μU/ml (immediately before the sustained infusion of exogenous bonito insulin) to 184±63μU/ml four hours after the start of the infusion. The calculated endogenous insulin output in the group treated with exogenous insulin also decreased significantly (p<0.05).
The two phases of glucose-induced insulin secretions in the insulin treated subjects were less marked than those in the control group (p<0.05), and occurred much more rapidly than those in the control.
These results suggest that the insulin secretion from the pancreas is affected by the prolonged infusion of exogenous bonito insulin.

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The stimulating effect of a spleen dialysate on the testicles and the accessory genital organs motivated the authors to test this dialysate on the ovaries. Also on the ovaries a different influence was observed.
In comparison to the control animals, the enzyme pattern: lactate dehydrogenase, steroid-3-β-ol-dehydrogenase, alkaline phosphatase, succinate dehydrogenase, and, glucose-6-phosphate-dehydrogenase is significantly changed. The effect is manifested by an increase of the tertiary follicles, increased follicular atresia and disquamating granulosa. On the basis of these findings we assume that the spleen dialysate is able to increase the gonadotropin effect on the ovaries.

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The metabolism of 3H-testosterone by the seminiferous tubules, prostate and muscle from immature rats was studied in short-time organ culture. The formation of the sum of ³H-5α-reduced products by the seminiferous tubules was similar to that by the prostate and was much higher than that by the muscle. ³H-5α-androstane-3α, 17β-diol was the main metabolite in the seminiferous tubules, while ³H-17β-hydroxy-5α-androstan-3-one was the main metabolite in the prostate. Following incubation of tissue explants with ³H-testosterone and immediate separation of the tissue from the medium, the seminiferous tubules contained relatively higher concentrations of the ³H-5α-reduced products and radioactive less polar products than 3H-testosterone or ³H-androst-4-ene-3, 17-dione. After hydrolysis of the radioactive less polar products with potassium hydroxide, 72% of the radioactivity was recovered as ³H-5α-androstane-3α, 17β-diol. These findings seem to indicate that the seminiferous tubules have a character of androgen-responsive tissue.

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Orchiectomy in rats resulted in retarded body growth, and an increase in thymus and adrenal weights. With concomitant increase in adrenal cholesterol and corticosterone contents, adrenal norepinephrine, a minor component of medullary hormones, and the total catecholamine contents were also increassed by the operation. These changes were prevented or to be recovered by 2mg of testosterone propionate supplement (1mg per rat per week for 2 weeks). A conventional estimation of the cortical and the medullary volumes revealed that orchiectomy caused a marked increase in the whole adrenal and cortical volumes and also an increase in the medullary volume, although it was not statistically significant. The data suggest that androgen in male rat affects not only the adrenal cortex but also the medulla.

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The human PTH radioimmunoassay has been developed by use of bovine PTH and anti-bovine PTH antiserum, both of which were commercially available. The iodination process was modified as to use acid solution resulting in the more stable ¹²⁵I-PTH. However, the PTH radioimmunoassay employed this antiserum with ¹²⁵I-bovine PTH revealed that the N-terminal portion of PTH molecule was reactive only slightly in this system, indicating that most of PTH determined by this radioimmunoassay might be an biologically inactive PTH.
Plasma from normal subjects and patients with primary hyperparathyroidism or uremia showed dilutional curves parallel to the curve of the standard bovine PTH, indicating the immunological identity between human and bovine PTH in this radioimmunoassay system. The extract from the parathyroid adenoma showed an identical dilutional curve, but the extract from the ectopic PTH producing tumor showed a different dilutional curve from that of the standard PTH, which might indicate the immunochemical heterogeneity of PTH produced from the ectopic source. Plasma PTH levels obtained from normal subjects and patients with primary hyperparathyroidism, uremia or pseudohypoparathyroidism were compatible with those reported previously. These data indicate that the human PTH radioimmunoassay can be done by use of commercially available anti-bovine PTH antiserum and bovine PTH in conjunction with the modified iodination process and double antibody method. However, this assay was directed mainly to the C-terminal portion of PTH molecule and the PTH radioimmunoassay specific for the N-terminal portion will be required to evaluate the PTH regulation more precisely in man.

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In the present study, two types of thyroglobulin-related proteins were solubilized from hog thyroid microsomes by sonication or detergent treatment and separated by polyacrylamide gel electrophoresis. The faster migrating protein had the same electophoretic mobility as that of soluble thyroglobulin.
These two proteins had the same immunogenicity as that of soluble thyroglobulin and lower iodine contents than the latter, suggesting a relationship between the proteins and newly synthesized molecules of thyroglobulin.

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