Reported here, are the observations on various histochemical enzyme reactions in several organs (the liver;the kidney;the cardiac, diaphragm, and femoral muscles;the duodenum;the adrenal gland) of rabbits which had been rendered diabetic for from 3 days up to 1 year or more. Independent of the diabetes-inducing reagents (Alloxan or Dithizone) used, the following similar results were obtained: With SD, LD and DPND (which are related to the Krebs cycle activity) and with such phosphatases as ACPase and ALPase, a distinct activity increase could be seen in the liver, the kidney, and the muscular system;except with SD, however, this increase tended to become indistinct or even slightly reduced when 1, 2, or 4 months had elapsed after the induction of diabetes. With G-6-PD (which is related to HMP cycle activity), a distinct decrease in the liver and the kidney could be seen throughout all the diabetic stages. The changes in Ph-rylase activity ran roughly parallel to those in glycogen content, but contradictory changes were found between activity and content in the lipemic heart. Regarding ATPase activity in the lipemic animals, a heavily stained membrane activity could be noted in the liver, the kidney and the cardiac muscle. The lipemic animals also revealed a remarkable increase in non-specific esterase. On the findings in the adrenals, the activities of G-6-PD, SD, ACPase, and ALPase had all been reduced, particularly in the fasciculata when diabetes had persisted for more than 2 or 3 weeks, and ran fairly parallel with the urinary 17-KS and 17-OHCS contents.
In addition to these findings, such interesting phenomena were noted as the tendency for the specific zonation of hepatic enzyme distribution to become obscure, the presence of an intimate relation between the activity changes in the liver and the kidney, and the presence of relatively specific activity distribution patterns in the various organs of rabbits having sustained experimental congenital diabetes for more than 1 year.

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Reported here, is an investigation of changes in enzyme activities in the pancreatic islet cells (A-and B-cells) from Alloxan or Dithizone-induced diabetic rabbits.In order to identify each cell type on the section stained for enzyme activity, the “Serial Section Method” was applied as follows: Two serial cryostat sections of the tissue were prepared: One of the sections was stained with chrome-hematoxylin phloxin for cell identification and the other was stained with various stains for enzyme activity. Then, the microscopic findings on the two sections were carefully compared. In the stained normal islets, the G-6-PD, ACPase, ATPase, LD, DPND, and SD activities were found to be almost the same in both the A-and B-cells, while the G-6-Pase activity was found to be very strong in the B-cells, in sharp contrast to its very weak reactivity in the A-cells.
In the diabetic islets, the activities of G-6-PD, ACPase, and ATPase showed a distinct reduction in both the A-and the B-cells, especially in the latter.No definite changes in G-6-Pase activity in the B-cells were found, as the changes varied from islet to islet or from cell to cell. However, many of the B-cell showed a reduced activity, but those B-cells which were found to possess comparatively large nuclei, and therefore seemed to have been regenerating, presented a stronger enzyme activity than the cells, which were found to possess small (pycnotic) nuclei or show marked degenerative changes. The A-cells showed little G-6-Pase activity. Regarding the changes in the activities of LD, DPND, and SD, nothing striking was noted.
The results obtained above reveal the presence of a fairly good proportional relationship between the activity changes in the A-cells and those in the B-cells.

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Human adrenal tissue from a fetus aborted after 6 month's gestation was sliced to 30μ thickness and incubated with estradiol-17β-16-¹⁴C in Krebs-Ringer bicarbonate buffer solution. The estriol fraction from the incubation mixture was purified by gradient elution partition chromatography on a Celite column, paper chromatography and thin layer chromatography to constant specific activity. Throughout these purification steps, the radioactive metabolite in the estriol fraction behaved exactly as the added carrier standard estriol. It was, therefore, considered that estradiol-17β-16-¹⁴C was converted to estriol by the incubation experiment with the fetal adrenal tissue. The overall conversion rate was 1.1%. In addition, the possible role of a fetus in the formation of estriol during pregnancy was discussed.

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The whole testis tissue preparation of rat or man was incubated with androst-4-ene-3β, 17β-diol (4ADL)(20 or 40μg in one flask containning 1 or 2 testes) for 2 hrs.in Krebs-Ringer bicarbonate buffer, pH 7.4, in 95% 0₂-5% CO₂ gas phase. A major product was proved as testosterone by means of paper and gas chromatographic analyses and by the derivative formation. Human chorionic gonadotropin (50 or 100 U in 1 flask) was shown to have a minor influence on the conversion of 4ADL to testosterone by rat testis in vitro.
Androst-5-ene-3β, 17β-diol (5ADL) was also converted to testosterone in the same rate as observed for 4ADL. Conversion of these 2 androstenedioles to androst-4-ene-3, 17-dione (4ADN) was observed to very minor extent compared to the conversion of dehydroepiandrosterone (DHEA) to 4ADN.
4-C¹⁴-DHEA was incubated with rat testis tissue preparation for 2 hrs. and 8 radioactive compounds converted from DHEA were separated on paper by extensive chromatography. Among these compounds, testosterone and 4ADN were identified by recrystallization with carrier steroid to constant specific activity. A compound behaving very similarly to the reference 4ADL on papers was separated. Oxidation of this compound with chromium trioxide yielded 2 products which showed the same mobilities as 4ADN and testosterone. An aliquot of the compound was added with carrier 4ADL and recrystallization showed the nearly constant specific activity. Testosterone zone derived from the compound by oxidation with chromium trioxide was added with carrier testosterone for recrystallization and the constant specific activity was demonstrated.

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The stress of injected epinephrine was reckoned from the reduction of ascorbic acid in the adrenal. In a young rat given epinephrine on day 3 after birth by spontaneous delivery, a depletion of ascorbic acid was not observed. Among similar young rats which were pretreated with hormones (LH, estradiol benzoate, testosterone propionate, progesterone, blood serum of pregnant rat and acetone-extract of rat placentae) and then given epinephrine, those of the group treated with LH and those of the group treated with extract of placentae showed significant depletions of the ascorbic acid in the adrenals. In newborns from spontaneous deliveries and in premature newborns obtained by Caesarean section, the weights of the adrenals declined during the first 24 hrs. ex utero-declined in fasted young and in nursed young.

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Apparent differences between the time curves (as determined by a method designated as the circulation method) of corticosterone concentration in the adrenal venous blood in hypophysectomized rats followed by the administration of natural ACTH and synthetic adrenocorticotropic peptides were demonstrated. When the areas under the curves obtained for 2 or 3 different doses of various adrenocorticotropic preparations were assumed as the responses, parallel dose-response regression lines were obtained for all the peptides tested. The relative potencies computed form this dose-response relationship somewhat differed from those by the established methods. It is suggested that the circulation method may be useful in evaluating the adrenal-stimulating potency of various preparations for therapeutic purposes.

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The amount of liver fat just before birth and just after birth in the rat was determined by weighing crude fat extracted with chloroform-methanol. The liver fat in rat fetuses tended to be decreased in amount toward birth. After birth, it was remarkably increased. In fetuses retained experimentally beyond the normal gestation, it showed a slight increase. In premature newborns obtained by Caesarean section, it was increased significantly in a short period of 6hrs. after the delivery.

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The study was made on the relationship between in vivo vaginal TTC reduction and the epithelial histology of mouse vagina under the influences of various antiestrogens such as MER-25, progesterone and testosterone. MER-25 markedly inhibited the estrogen-stimulated vaginal TTC reduction when administered simultaneously, but the inhibition was reduced or abolished according to the injection interval between estradiol and MER-25 being prolonged for 3, 6 or 12 hrs. Progesterone and testosterone, however, produced almost consistent inhibition without regard to the time of administration within a period of 12 hrs. after the estrogen injection. Histological investigations on the vaginal epithelium also demonstrated that MER-25 almost completely blocked an earlier response to estrogen, primary proliferation of epithelial cells, while progesterone and testosterone allowed the epithelial proliferation in a relatively normal manner. These results indicate that MER-25 only inhibits earlier step of estrogen action, whereas either progesterone or testosterone inhibits rather later stage. It is found that there is close correlation between the ability of formazan production in the superficial cells and differentiation of vaginal epithelium under influences of these anti-estrogenic substances, supporting the previous explanation for the mechanism of hormonal regulation of vaginal TTC reduction.

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