Polyglucose synthesis was investigated by incubating the rat skeletal muscle in vitro using the media containing glucose-1-phosphate under the electron microscope and the results were compared with those of its iodinestaining under the light microscope. In the usual skeletal muscle, apart from the soleus muscle, polyglucose of α-1, 4-linkage with longer outer chain stained blue with iodine and it appeared in large white fibers. It revealed an amorphous aggregation containing a finer granular appearance with a diameter from 100Å to 150Å under the electron microscope. Such polyglucose deposition appeared to be in the subsarcolemmic and sarcoplasmic matrices of these fibers, which agreed with Takeuchi and Sasaki's findings. The branched polyglucose of α-D-1, 4-linkage with shorter outer chain, stained red with iodine, was demonstrated as electronmicroscopically larger particles with a diameter from 550Å to 800Å (average 632Å), which were most predominant in number and deposited not only in the subsarcolemmic matrices but also in the sarcoplasmic ones of the small red fiber of white muscles. Polyglucoses varied in a particle size from 250Å to 450Å in diameter were predominantly synthesized in the intermediate fiber of the white muscle and they were stained violet with iodine histochemically. These particles, different in size and density from native glycogen particles, appeared only by the electron histochemical procedure. They were stained with varied density by lead staining under the electron microscopic technique, compared with the native glycogen particles. However, there were sometimes some patterns of different areas occupied by small groups of newly formed polyglucose particles, which were different in their particle size and electron density in each localized area.

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In an attempt to elucidate the nature of ruthenium red reactions of substances in histochemical specimens, the effects of added electrolytes upon the reactions of model polyanions on filter paper and in tissue have been studied according to the Scott-Dorling type of critical electrolyte concentration (CEC) principle. The results obtained in the present experiments indicate that any of the three anionic groups, sulfates, phosphates and carboxyls can be responsible for the ruthenium red reactions of polyanions and that the dye binds to polyanions largely forming a salt type of linkage. However, ruthenium red is not a dye of choice for use in stainings based upon the CEC principle, inasmuch as certain moieties of the dye cations appear to interact with cationic electrolytes.

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Mucosal glycoprotein was isolated from human intestinal mucosa. It possessed higher sialic acid and sulfate contents than those of the gastric mucosal glycoprotein. The former was, therefore, stainable with Alcian Blue while the latter was not. These two kinds of glycoproteins gave a specific precipitate line against the corresponding antiserum, respectively, on the Ouchterlony assay. Using the direct immunofluorescent antibody technique, the intestinal glycoprotein was demonstrated to distribute in the alcianophilic goblet cells throughout the intestinal mucosa but only faintly demonstrable in the surface epithelium of gastric mucosa.

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The effects of folate and methotrexate on dihydrofolate reductase, the activity of some dehydrogenases, and on DNA quantity were studied in “in vitro” cultured human connective, epithelial and HeLa cells. Normal or cancerous epithelial cells seemed more sensitive than fibroblasts to the addition of folate or methotrexate.

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Recently it has been stressed that catecholamines play a significant role in the pathogenesis of hypertension in Spontaneously Hypertensive Rats (Okamoto and Aoki) (SHR). In this report electron microscopical observation in male SHR was made on the adrenal medulla through the processes of the hypertension. The mild hyperfunctional state of norepinephrine-secreting cells in the initial stage of hypertension was expected on the feature of increased population of norepinephrine-secreting cells, its prominent Golgi complex and rough-surfaced endoplasmic reticulum and increased degranulation in the cell. Moreover, it may be assumed a certain functional change of epinephrine-secreting cell system on the base of increased population of dark epinephrine-secreting cells which are regarded as premature cells.

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The distribution and localization of arginase in the bovine blood cells was studied by immunofluorescent method. Specific antibody against arginase by immunizing rabbit with the corresponding antigens was conjugated with fluorescein isothiocyanate.
The fluorescence was intensely demonstrated in the reticulum cell and immature cell (erythroblast, promyelocyte, etc.) by microscopic examination of the bone marrow cells of the bovine. Slight arginase was recognized in the erythrocyte and lymphocyte, no arginase in the monocyte and polymorphonuclear neutrophilic leukocyte. Arginase distributed diffusely in the cytoplasm of above cells. It was never found in the nucleus.
Immunoproperties of fluorescent antibody was investigated by gel chromatography on Sephadex G-200 or immunoelectrophoresis. Physiologic significance of arginase in the blood cells was also discussed.

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