By means of acetone precipitation, a substance was prepared from the fraction obtained by removing parotin from the autolyzed bovine parotid gland. Its biological properties were similar to parotin possessing the hypocalcemic and leucocytosis-producing activities for rabbits. This parotin-like substance was named β-parotin.
Autolysis at 37°C for 24hrs, was most effective for preparing β-parotin. The parotin obtained from the autolyzed gland was also active in both effects.

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Acetone fractionation, acrinol treatment and preparative zone electrophoresis were tried to isolate, β-parotin, and it was clarified that the zone electrophoresis was an excellent procedure for the separation of highly purified β-parotin. And, simultaneously, a by-product, which was considered to be parotin, was separated.
The isoelectric point of β-parotin was estimated as pH4.2 by using paper electrophoresis, and that of by-product was roughly estimated as pH5.5 to 5.7 from its solubility curve using a buffer solution.

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Crude and purified β-parotin obtained from fresh parotid gland were biologically inactive, while the same products obtained after the autolysis of the gland were active. However, it was found that the inactive material could be easily activated by the incubation at 37°C; the most suitable condition was 24 to 48hrs. incubation at pH 5.5 in an aqueous solution.
Although the mode of activation is still remained to be clarified, it seems to occur without an activator, because the activation was observed even with a highly purified material.

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1. The ovarian weights and the ovarian ascorbic acid contents of immature female rats after pretreatment with gonadotrophins (PMS 75 i.u. and HCG 30 i.u.) were compared between 4 strains of rats. The ovarian weight was the largest in Holtzman strain, next in Wistar-Imamichi and Sprague-Dawley (Takeda) strains, and the smallest in Wistar strain. The ovarian ascorbic acid content was the highest in Holtzman strain, next in Wistar-Imamichi and Sprague-Dawley (Takeda) strains and the lowest in Wistar strain.
2. It was found that the OAAD method would produce the best result when used by 3-hour-method in Holtzman strain, and by 2-hour-method in Wistar-Imamichi strain.
3. In the dose response curve of NIH-LH-S1, the indexes of precision (lambda) of Holtzman and Wistar-Imamichi strains were excellent, being 0.1686 and 0.1650 respectively, and lambda of Wistar strain was inferior, being 0.2309 to 0.500. MED was 0.2μg in Holtzman strain, 0.5μg in Wistar-Imamichi and Sprague-Dawley (Takeda) strain, and 2.5μg in Wistar strain.
4. The OAAD method proved specific to the hormone with LH activity.

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1. Human serum LH levels were assayed by using ovarian ascorbic aci depletion method. The 4 point assay using 2 doses of NIH-LH-S₁ and of huma serum demonstrated parallelism. Human serum failed to augment LH activit in ovarian ascorbic acid depletion test.
2. The human serum LH level of normal menstrual women showed the peak in the preovulatory phase. Ovariectomy in women considerably increase serum LH level.

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Adult male Wistar-Imamichi rats were given the drinking water containing 1-2 ppm or 200 ppm of iodine ad libitum for 5, 15 and 60 days. The thyroid gland cells were investigated light and electron microscopically. The mean value of T/S ratio was lower markedly in rats with 200 ppm of iodide intake. The peculiar big canals invaginated deeply the gland cell from the colloid lumen in most of rats given 200 ppm of iodine for 5 and 15 days, whereas they were not recognized in rats given for 60 days, therefore the occurrence of them is temporal. These canal systems extend irregularly and are complicated in shape, either dilated or constricted. The cross-sections of the canals represent huge granules and are eventually in contact with the basement membrane, invading the deepest area. The content of the canals seems to be identical with the luminal colloid and their wall is equipped with the elongated numerous microvilli. The intracellular canals in thyroid gland cells have been observed for the first time in the present electron microscopy. As to their functional significance, the possible transient reabsorption of thyroglobulin has been suggested.

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The ultrastructures of cells of the anterior lobe in normal and castrated male rats, 50, 128 and 300 days after the removals were investigated.
1. Chromophobe cell is a maternal or immature cell from which acidophil and basophil may develop. The transformation axis from chromophobe cell to basophils was exclusively stimulated in degree by the castration. Following the castration, granulation was progressive in the enlarged cytoplasm of chromophobe cells, which may turn into polygonal or round basophils.
2. The size-difference in dense beta-granules in basophils is not indispensable for the identification of gonadotroph and thyrotroph, because beta-granules, 100 to 150mμ in diameter, in immature basophil (thyrotroph-type) of castrated rats may grow up to large one, 200 to 250mμ in diameter, which are common with those in chronic castration cells. Thus the castration made the number and the size of granules in basophils increase. Polygonal thyrotroph may correspond to a cell preceding to round gonadotroph in a series of life history of the basophile cell.
3. Beta-and alpha-granules may be produced together in Golgi area. Evidences were presented that vesicles were gradually transformed into granules of variable density. The former, therefore, assumed to be a precursor of the latter. The present observation indicated that granules were stable vehicles for the longterm preservation in the cytoplasm, and vesicles were seemingly important carriages which might work as the active principle.
4. Following the chronic castration, most of basophils turned into the cast ration cells which were thereafter converted into signet-ring cells, in which huge colloid lumen seems to be originated from the coalescence of smooth endoplasmic reticulums. Within 128 and 300 days, signet-ring cells were provided with a degenerating sign on account of prolonged preservation. Big granules (700 mμ) yielded numerously.
5. In addition to alpha-granules, so-called prolactin granules with ellipsoid contour are rarely released out of cell-body by adhesion of the limiting membrane of the granules to the plasma membrane even in male rats. They may not be pushed out but mostly dissolved by enzymatic activity. On the other hand, the discharge of beta-granules presumably takes place in normal and castrated rats by diffusion of the granular content due to the vanishment of the limiting membrane. Diffused material might be eliminated through the membrane of endoplasmic reticulum and stored in it.
6. The above ultrastructures would reinforce the previous doubt of the authors about the subdivision of gonadotroph and thyrotroph.

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