Histochemical observations on mucosubstance are studied in total 270 specimens which are composed of 200 specimens (or 265 tissue blocks) of gastric carcinomas, 50 specimens of non-cancerous gastric tissues, and 20 specimens of duodenal and rectal tissues. The results are summariezd as follows: 1. In the normal and non-cancerous gastric mucoas, two types of mucosubstance are distinguished histochemically: Mucosubstance proper to gastric mucosa (stomach type or S-type) being stained various tints of red in colour by AB-PAS method (PAS-positive and AB-negative), and mucosubstance specific for goblet cells in the area of intestinal metaplasia (goblet type or G-type), being stained blue to purple by AB-PAS method (positive for both AB and PAS). 2. The incidence of G-type mucosubstance in gastric carcinoma tissues is unexpectedly high, amounting to 157 out of 200 specimens or 78.5 per cent; or 212 out of 265 blocks or 80.0 per cent. 3. The incidence of G-type mucosubstance in the early (intramucosal) carcinoma of the stomach is 51/65 (78.5%), and this is quite similar to that in the advanced carcinoma, 106/135 (78.5%). 4. According to these findings, a close relationship of G-type mucosubstance in the gastric carcinomas to the goblet cells of intestinal metaplasia is assumed, and a possible roll of the intestinal metaplasia in a histogenesis of the gastric carcinoma is considered.
The localization of both diamine oxidase and D-amino acid oxidase was examined by immunofluorescent technique on the frozen section of the porcine kidney. The cells, composing the Bowman's capsule and the proximal convoluted tubule, showed intense fluorescence which showed the localization of diamine oxidase. The antigen was concentrated in the supra- and para-nuclear cytoplasm. But, unlike monoamine oxidase, diamine oxidase may not be located in the mitochondria, because higher concentration of diamine oxidase is found in the apical cytoplasm where the mitochondria are not so much observed. The D-amino acid oxidase was demonstrated in the apical cytoplasm of the cells constituting the Bowman's capsule, the proximal convoluted tubule and a part of the Henle's loop. The place where two enzymes were located was, as mentioned above, mostly in the proximal convoluted tubule. At this place, the large amount of ammonia was reported to be secreted and the highest activity of the two enzymes was also encountered. In other words, the location of two enzyme proteins, the place where ammonia is eliminated and the sites of enzymatic activity of two enzymes were quite identical.
The localization of ovalbumin in the chick oviduct was investigated by means of fluorescent antibody technique. The infundibulum, oviductus proprius and isthmus had serous gland which secrets egg-albumin in the spawning-time. Since the oviductus proprius was the longest of three parts of the oviduct, and its serous gland was most well developed, specific fluorescence in the serous gland was found most frequently in this part. On the contrary, even in the spawning-time, the other two parts showed little specific fluorescence. The volume of the secretion of egg-albumin changed according to the descending stage of the egg. Namely, the secretion of egg-albumin increased as eggs descended, and then, secretion of serous gland ceases gradually after the eggs passed down. Besides the decrease of the height of each mucous epithelial cell, specific fluorescence was not found in every region of the three parts of the chick oviduct, during the resting time of ovulation.
The distribution of arginase in the bovine liver was investigated by aid of the fluorescent antibody method. The crystalline arginase used in this study was found to be composed of five components by the Ouchterlony's gel diffusion test. This enzyme preparation was separated into five or six major fractions by the gel filtration method with Sephadex G-200. One of these fractions was attributable to bovine serum albumin, and the other two were common with the components of bovine heterogeneous organs and the liver of heterogeneous animals. Consequently, this enzyme preparation was composed of two components from the immunological point of view. The enzyme is encountered mainly in the liver cells surrounding the centralvein and additionally in the cells of the indefinite area of a liver lobule. In the cytoplasm of each liver cells, the enzyme distributes itself diffusely. It is never found in the nucleus. Furthermore, the specific fluorescence is not seen also in the endothelial cells of the sinusoid, Kupffer's cells, the cells of the wall of the interlobular bile duct and in the endothelial cells of the interlobular blood vessels. The presence of arginase in the bovine kidney was not recognized in this study, though it has been already proved biochemically. Moreover, the specific fluorescence has not observed in the liver and kidney of the ureotelic animals, pork and mouse, though the extract of these organs produced several precipitin lines against anti-bovine liver arginase serum on the Ouchterlony's plate. Furthermore, the specific fluorescence has not been observed in the liver and kidney of the uricotelic animal, the chick. From these findings, it is supposed that the bovine arginase have high species specificity as well as high organ specificity.
The localization of aminopeptidase was examined on the porcine intestine by use of fluorescent antibody method. Aminopeptidase was present diffusely in the striated border of mucous epithelial cell of jejunum and ileum, supra-nuclear region of epithelial cell, intestinal glandular cells and in goblet cell. The condensed materials in the crypts showed specific fluorescence. The antigen was not always observed in every cell mentioned above, and the distribution pattern of antigen varied according to the stage of digestion. The enzyme was concentrated in the goblet cells of the small intestine but not in those of the large intestine. Brunner's gland of duodenum did not contain any aminopeptidase. Though aminopeptidase in kidney and pancreas had immunological common factor with that in the intestine, specific fluorescence was not observed on the tissue sections. For comparative purposes, histochemical staining was carried out by means of Burstone and Folk's method. As a result, the goblet cell of the small intestine did not show any cytochemical reaction, though the specific fluorescence which showed the site of the enzyme protein was concentrated in it. As far as the striated border of both mucous epithelial cell and intestinal glandular cell is concerned, the results of the cytochemical reaction coincided with those obtained by immunohistochemical study. The reason for the discordance with regard to the distribution of cytochemical reaction and the immunofluorescence in the goblet cell remained unknown.
Intracellular distribution of polyglucose synthesized histochemically from glucose-l-phosphate by phosphorylase in rat skeletal muscle was examined on electron microscopic level. Newly formed polysaccharide bearing specific properties appeared in a granular form finer and less dense than the original glycogen particulates of the muscle bfiers, as described in detail in our previous report. It was in general deposited abundantly in intermyofibrillar spaces and more abundantly in subsarcolemmic areas. The synthesis occured in sarcoplasmic ground substance of these areas frequently showing very close localization with native glycogen particulates remaining in the muscles without resolved into the substrate mixture. Among various organelles in the muscle fibers containing polyglucose, sarcoplasmic reticulum received the most conspicuous influence in its arrangement by abundant deposition of the reaction product. Especially, in subsarcolemmic areas, it was remarkably expanded by the polyglucose accumulation and it divided the areas into many subareas. Whorls of sarcoplasmic reticulum membranes were occasionally observed in polyglucose areas. Nuclei, mitochondria and myofibrils did not reveal any direct relationship with polyglucose. On the basis of these observations, intracellular localization and distribution of phosphorylase which plays an active role in this reaction was discussed.
LDH and MDH were studied histochemically in the normal and regenerat-ing tail of the lizard, Mabuya carinata. This study revealed an identical pattern of localization and distribution for these two enzymes during the various stages of regeneration. A slight initial fall in activity during the earlier phases was quickly followed by a gradual rise of activity during the later phases of regeneration. The high activity of MDH alongwith LDH in the light of hitherto reported low activities of SDH and ICDH is indicative of a strong pyruvate centered metabolism with lactate, oxaloacetate and malate acting as important coparticipants. The revelations of this investigation tends to indicate the existance of a short cycle similar to that of pyruvate shuttle reported in vertebrate liver and muscle. This cycle has the advantage of supplying NADPH2 an important cofactor for lipogenesis. Further significance of this cycle is discussed.