Primarily the cytochrome c-leucopatent blue-hydrogen peroxide (CC-LPBH₂O₂)reaction intensely stained the matrix of cartilage, substantia propria of the cornea, and mast cell granules, and occasionally stained the nuclei of certain epithelial cells. In the nuclei, non-specific combination of CC with various components was dissociated by treatment with buffer solution of pH 3.0. After methylation, the reactions of mast cell granules(37°C), substantia propria of the cornea(37°C)and cartilage(60°C)were entirely abolished. In sulfation sections, the reactions of mast cell granules were completely reduced, while those of goblet cells, collagen fibers and arterial wall were markedly enhanced. By digestion with novel hyaluronidase, the reactions of umbilical cord and cockscomb were abolisned, while mucopolysaccharides containing sulfuric acid esters were completely uninfluenced except in the case of the cornea and sclera. In digestion tests with chondroitinase ABC and AC, mast cell granules were completely uninfluenced, while cartilage and mucous acinous cells of the lingual glands retained their staining affinities with CC-basic proteins to some extent. In the critical electrolyte concentration method for distinguishing individual acid mucopolysaccharides, the staining pattern shifted and became strikingly reduced compared to the Alcian blue method. Mast cell granules and tracheal cartilage, however, were clearly stained at 0.2 M MgCl₂in CC-adjective solution in contrast to staining of the cornea at 0.05 M MgCl₂. The CC-adjective reaction is thought to provide a method for the localization of polyanions such as sulfate, carboxyl and phosphate groups. In the specific reaction at pH 3.0, however, its staining property generally runs parallel with the acidity and density of the polysulfated mucopolysaccharide. Further, data obtained by histochemistry were in agreement with those obtained by electrofocusing analysis of CC-heparin complex in vitro.

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The authors have investigated the structure, and mucopolysaccharides histochemistry of the submandibular glands of man, Japanese macaque, dog, cat, tree-shrew, Eastern mole and rabbit, and concluded as follows:
1. The submandibular glands of man, Japanese macaque, dog, cat, tree-shrew and Eastern mole are composed of two kinds of acinar cells while the rabbit has only one kind of cell.2. The cells near the intercalated duct in the rabbit submandibular gland (the socalled Ichikawa's B cells) do not belong to the glandular cells but presumably have their origin in the intercalated duct cells. Some scholars called the same duct as the granular intercalated duct.3. The intercalated duct cells in the rabbit are different from the granular duct cells of the Rodentia in their locations, but both of these cells have close similarity in their histological and histochemical characteristics and electron micrograph 4. Excepting Japanese macaque and Eastern mole, the neutral, weak and strong acid mucopolysaccharides are provable in two kinds of glandular cells of the submandibular glands observed in the present study. Strong acid mucopolysaccharide is improvable in the A cells of the Japanese macaque and both the A and the B cells in the Eastern mole.5. It is impossible to classify the glandular cells by the mucopolysaccharide histochemistry alone. Depending on the histological, histochemical and electron microscopical characteristics the authors regard that the distally located cells (A cells)in the mixed acinus belong to the seromucous cells, and that the cells near the intercalated duct (B cells) to the mucous cells.6. Clear protoplasm, flattened and basally located nucleus and no secretory capillary are the characteristics of the mucous cells (I type of mucous cells). The B cells of the tree-shrew are dark and their nuclei are round or oval. The same cells of the Eastern mole are clear and their nuclei are flattened and basally located, but have scanty mucopolysaccharide. In spite of the above-mentioned characteristics, the authors regard that both the B cells of tree-shrew and Eastern mole belong to the II type of mucous cells by reason of the lack of the secretory capillary.7. The core in the secretory granule is one of the characteristic of the seromucous cells, but the same granules of the dog have a few small vesicles for the core, and the granules of the Eastern mole exhibit a peculiar pattern and have some resemblance to those of the mucous cells (B cells)in the tree-shrew and the so-called serous cells of the European hedgehog except the core of high electron deuce in the granules.8. The mucous cells are filled with large secretory granules having no cores but containing abundant fine particles. On the other hand, the same granules of the tree-shrew and those in the so-called serous cells of the European hedgehog corresponding to the mucous cells of the tree-shrew exhibit extremely characteristic patterns, and both of the patterns are very similar in their characteristics. The said patterns do not appear in the seromucous and mucous cells of the submandibular glands of Primates and Carnivora.9. The reactions to neutral, weak and strong acid mucopolysaccharides in the A cells of the rabbit are all very clear and many protein reactions are negative. These characteristics are peculiar to the mucous cells. Moreover, the electron micrograph of the secretory granules in these cells are extremely similar to that of the mucous cells. Nevertheless, the authors classify these cells under the category of the seromucous cells owing to the presence of the secretory capillaries in the same cells.10. Neutral and weak acid mucopolysaccharides are regular substances in the acinar cells of many kinds of the salivary glands of man and various kinds of mammals. Some kinds of acinar cells in several animals lack the strong acid mucopolysaccharide but the authors do not approve of the existence of the serous cells in the salivary glands.

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The effect of protein deficiency on lymphoid and other organs in CF #1 mice of both sexes was studied histologically and cytochemically. The effect of refeeding a regular diet after feeding a protein-free diet, and the effect of starvation on the lymphoid organs, were also investigated.
The mice were initially fed a protein-free diet and a regular diet (25% casein)at 21 days of age. The weight and number of nucleated cells in the lymphoid organs were significantly lower in the protein-deficient mice than in the controls. This damage to the lymphoid organs was not a permanent one and could be remedied after an adequate period on the regular diet.
The content of naphthol yellow S-total protein in the small lymphocytes of the thymus and the lymph nodes of the protein-deficient mice was very similar to that of the controls. The contents of Feulgen-DNA and fast green histone in the lymph node lymphocytes of the protein-deficient mice were almost the same as those in the control mice.
The weights of the adrenals and gonads in the protein-deficient mice and starved mice were large, if expressed as weight per body weight, but no definite histological changes were observed. Thus, lymphoid hypoplasia may be related to the presence of intact adrenals and gonads in protein-deficient mice.

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