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Biomedical subjects

F Murata

Publications and source records attributed to F Murata.

At least 73 records · Page 4Linked to original sources
PubMed1982

Post-embedding staining of rat gastric mucous cells with lectins.

The mucous cells of the rat stomach were stained with lectins by two post-embedding staining methods for electron microscopy. The mucous granules of surface mucous cells and foveolar mucous cells were stained weakly by Ricinus communis agglutinin-ferritin and wheat germ agglutinin-ferritin. The mucous granules of mucous neck cells were stained by concanavalin A-ferritin, Ricinus communis agglutinin-ferritin and wheat germ agglutinin-ferritin. The mucous granules of pyloric gland cells showed an affinity for wheat germ agglutinin-ferritin and concanavalin A-ferritin, while Ricinus-communis agglutinin-ferritin only slightly stained the granules. The granules of mucous neck cells and pyloric gland cells were also stained by the concanavalin A-horseradish peroxidase-colloidal gold method, but the granules of surface and foveolar mucous cells were not stained by this method. Periodic acid oxidation of the sections before the standard concanavalin A-ferritin procedure enhanced the staining of the granules of mucous neck cells and pyloric gland cells slightly. Reduction of the sections after the periodic acid oxidation weakened the staining. Similar results were obtained using the concanavalin A-horseradish peroxidase-colloidal gold method. Though the staining with Ricinus communis agglutinin-ferritin was inhibited by periodic acid oxidation of the sections before staining, the staining with wheat germ agglutinin-ferritin was not inhibited by the oxidation. It is suggested that the paradoxical staining is closely related to the position of the concanavalin A-binding sugar residues in the carbohydrate chains.

Animals↗
PubMed1982

Nucleic acid synthesis in proliferating peroxisomes of rat liver as revealed by electron microscopical radioautography.

Thirty albino rats were fed with a diet containing 1, 2 or 4% di-(2-ethylhexyl)-phthalate (DEHP), a peroxisome-proliferating agent. Others were fed with normal diet as controls. Both groups were sacrificed at varying intervals from 3 days to 4 weeks. The livers were either removed and fixed in glutaraldehyde and osmium tetroxide or fixed in glutaraldehyde, incubated in a diaminobenzidine (DAB) medium, postfixed, embedded in Epon, and sectioned. Other tissues were incubated in Eagle's MEM containing either [P3H-thymidine or [3H]uridine, fixed, embedded in Epon, sectioned, and radioautographed. Specimens were observed in a Hitachi H-700 electron microscope. The number of peroxisomes showing DAB reactivity increased in DEHP-fed animals as compared with normal controls. In radioautograms of normal rats labelled with [3H]thymidine, no silver grains were observed, whereas grains were observed overt some nuclei, mitochondria and peroxisomes of DEHP-fed animals. In contrast, radioautograms of tissue labelled with [3H]uridine revealed a few grains in nuclei and mitochondria or endoplasmic reticulum of normal rats, although grains appeared in nuclei, mitochondria, endoplasmic reticulum and peroxisomes of DEHP-fed animals more frequently. From these results, it is concluded that [3H]thymidine and [3H]uridine were incorporated in the proliferating peroxisomes, suggesting that nucleic acid synthesis had taken place.

Animals↗
PubMed1981

Comparative histochemical study of alimentary tracts with special reference to the mucous neck cells of the stomach.

Combinations of recently developed paradoxical concanavalin-A staining (PCS) and other routine histochemical procedures have made it possible to classify mucosubstances more precisely. By taking advantage of these sequences, the present study was undertaken both to characterize the mucinous contents of alimentary tracts of several animal species, and to compare the variability of epithelial mucosubstances with special reference to the mucous neck cells of the stomach. The alimentary tracts obtained from 7 species of fish, 5 amphibians, 3 reptiles, 4 birds, and 6 mammals were used. Acidity of epithelial mucosubstances was highly variable even among the corresponding mucous cells. On the contrary, concanavalin-A reactivity seemed to be consistent with each cell and species. The mucous neck cells, which were observed in amphibians, reptiles, and mammals, consistently exhibited a characteristic mucosubstance with stable class-III reactivity by PCS. Inversely, stable class-III reactivity was found only in species possessing mucous neck cells, and was widely distributed in the esophageal glands of the frog, snake, and man; in esophageal mucous cells of the frog and skink; in pyloric glands of amphibia, reptiles, and mammals; and in Brunner's glands of mammals. These mucous cells seem to form an unique group with morphological and histochemical similarities. It is likely that the stable class-III reactivity by PCS is a hallmark of the mucous neck cells and related glands and that, in addition, concanavalin-A reactivity of mucosubstances is evolutionarily more fundamental characteristic.

Amphibians↗
PubMed1981

Ultrastructure and immunohistochemical staining of a transplanted endodermal sinus tumor.

An endodermal sinus (yolk sac) tumor was successfully transplanted into athymic nude mice. Histologic and ultrastructural investigations revealed that the transplanted tumor had a characteristic appearance with numerous Shiller-Duval bodies, endodermal sinus structures and ultrastructural profiles as previously described in human material. The endodermal sinus tumor and normal human yolk sac have been found to synthesize not only alpha-fetoprotein (AFP), but also other serum proteins, namely, albumin, prealbumin, alpha 1-antitrypsin, and transferrin. Serological study by radioimmunoassay demonstrated AFP, carcinoembryonic antigen (CEA) and human chorionic gonadotropin (HCG) in the sera of the tumor-bearing nude mice and in cyst fluid from the transplanted tumor. Immunohistochemical investigation using the unlabeled antibody peroxidase-antiperoxidase method showed using the unlabeled antibody peroxidase-antiperoxidase method showed that the tumor cells produced CEA, alpha 1-antitrypsin, transferrin, HCG as well as AFP. These immunohistochemical staining properties were correlated with the findings on radioimmunoassay.

Adult↗
PubMed1981

Postembedding staining of Brunner's gland with lectin-ferritin conjugates.

Postembedding staining of intracellular carbohydrates of rat Brunner's gland cells embedded in Epon and acrylamide was carried out with Ricinus communis agglutinin-ferritin, concanavalin A-ferritin, and wheat germ agglutinin-ferritin conjugates. Th Golgi vacuoles and mucous granules were stained with these conjugates. In each staining, the tissues embedded in acrylamide were stained more strongly than those embedded in Epon. The staining intensity of wheat germ agglutinin-ferritin was the strongest among the three conjugates and the staining intensity of Ricinus communis agglutinin-ferritin was stronger than that of concanavalin A-ferritin in both embedding methods. Free ferritin showed almost no binding to these structures and staining with the conjugates was inhibited by the addition of appropriate competitive sugars to the staining solutions. Osmium-postfixed tissues were not stained well with the conjugates. Washing of the sections with bovine serum albumin solution after staining was an essential step in the present method to reduce the nonspecific adsorption of the conjugates. The present method was very simple and had good reproducibility.

Animals↗
PubMed1980

Immunoradioautographic study of alpha-fetoprotein in hepatoma cells.

Trypsin treatment, which is frequently used in the field of immunocytochemistry to faciliate conjugated antibody penetration through the cell membrane, was applied for the immunoradiographic demonstration of alpha-fetoprotein in ascites hepatoma cells using 125I-labeled anti rat alpha-fetoprotein antibody. No significant number of silver grains were detectable in the group without trypsin treatment, while numerous silver grains were recognizable in the ascites hepatoma cells after trypsin treatment. This indicates that trypsin treatment is effective for the immunoradiographic demonstration of alpha-fetoprotein and contributes to the quantitative immunoradioautographic investigation.

Animals↗
PubMed1979

Mucosubstances of rabbit granulocytes studied by means of electron-microscopic radioautography and X-ray microanalysis.

Rabbit bone marrow cells have been studied by means of light- and electron-microscopic radioautography and X-ray microanalysis. Carrier free sulfuric acid was used as the radioactive precursor in this experiment. Immature granulocytes showed more active incorporation than mature ones. Silver grains were observed in the Golgi apparatus and granules in three kinds of granulocytes. Electron-microscopically, immature granules showed the incorporation of inorganic sulfate, while mature ones did not. Sulfur was detected in all kinds of granules of the three granulocyte types by X-ray microanalysis. It is concluded that incorporated inorganic sulfur may be utilized for the synthesis of acid glycosaminoglycans and the sulfur detected by X-ray microanalysis may be that contained in the acid glycosaminoglycan. The sulfur detected in the specific granules of the heterophil probably derives from proteins or polypeptides incorporating sulfur containing amino acids.

Animals↗
PubMed1978

Ultrastructural localization of glycogen in the granulocytes of normal rabbit bone marrow.

The glycogen of rabbit granulocytes has been studied in glutaraldehyde and osmium tetroxide fixed bone marrow by the periodic acid-thiocarbohydrazide-silver proteinate procedure (PA-TCH-SP). The PA-TCH-SP procedure involved the staining of intracytoplasmic glycogen more densely than the routine lead citrate staining. The PA-TCH-SP procedure demonstrated the intracytoplasmic glycogen in all three kinds of granulocytes. Though a sequence of intensity was observed in each stage of cell maturation, intracytoplasmic glycogen increased generally in accordance with cell maturation in the granulocytes. Functional significance of the glycogen in the granulocytes was discussed in relation to its staining. A very weak reaction in the granules of the granulocytes was described in relation to their contents.

Animals↗
PubMed1978

A modified wire-loop method for quantitative electron microscopic radioautography.

A simplified method of emulsion coating and development for quantitative electron microscopic radioautography without any special instrument in introduced. Seven grids carrying ultrathin sections are attached to a glass block with double-coated Scotch tape. They are coated with a wire-loop, 2.5 cm in diameter. Ten glass blocks are then stuck on a slide, exposed, developed, fixed and stained simultaneously. Employing this method, the emulsion coating and the following procedures are easily accomplished under an identical condition.

Autoradiography↗
PubMed1978

Histochemical studies of mucosubstances in metaplastic epithelium of the stomach, with special reference to the development of intestinal metaplasia.

Processes in the development of intestinal metaplasia of the stomach were investigated from the morphological and histochemical approaches using light and electron microscopic techniques. The specimens taken from 38 gastric carcinomas and 15 gastric and/or duodenal ulcers were subjected to this study. Morphological appearances of the intestinal metaplasia observed in routine examination with hematoxylin and eosin staining was able to divided into complete and incomplete metaplasia by the light and electron microscope histochemical stainings of the mucosubstances. The columnar cells at the area of the incomplete metaplasia had both the properties of the intestinal epithelia and the gastric foveolar epithelia. The incomplete as well as the complete metaplasia arose from the generative cells at the isthmus of the gland. The generative cells, however, sometimes gradually transformed to produce the complete metaplastic cells. The two processes of the development of the intestinal metaplasia were proposed and discussed.

Duodenal Ulcer↗
PubMed1978

Electron microscopic observation of scirrhous carcinoma of the stomach.

Electron microscopic findings obtained from seven scirrhous carcinomas of the stomach were analyzed dividing each carcinoma into superficial, deep, and peripheral parts. Proliferation of collagen fibrils, disappearance of plasma membranes, and extracellular release of cell organellae of cancer cells were considered to be peculiar findings to this type of cancer. These phenomena were observed frequently at the deep part of the cancer, but rarely at the peripheral and superficial parts. Replacement of damaged cells by the proliferated collagen fibrils at the central area of the cancer and disconnection of cancer cells from scirrhous lesion at the peripheral part were considered to contribute to the biological behaviour of this carcinoma.

Adenocarcinoma↗
PubMed1977

Fine structural and ultracytochemical studies on the lymphocytes in three types of genetic mucopolysaccharidoses.

Lymphocytes from 6 patients with 3 types of genetic mucopolysaccharidoses (Hurler's syndrome, Hunter's syndrome and Morquio's syndrome) contained numberous vacuoles in their cytoplasm. The size of the vacuoles ranged from approximately 300 nm to 750 nm. The percentage of the lymphocytes with vacuoles varied from 10% to 38%. The vacuoles showed acid phosphatase activity, which indicated their lysosommal nature. Staining with dialyzed iron solution usually localized acid mucosubstance in the peripheral region of these vacuoles after glutaraldehyde fixation. Ferritin and horseradish peroxidase were observed in the vacuoles after incubation of the patient's lymphocytes with these tracers. This finding indicates the participation of endocytosis in the formation of these vacuoles.

Acid Phosphatase↗
PubMed1977

Electron microscopic dry-mounting radioautography for diffusible compounds by means of ultracryotomy.

Cultured HeLa cells or mouse liver and pancreas tissues were labeled with 3H-thymidine, -uridine or -glycine for varying periods in vitro, frozen in liquid nitrogen and cut on an LKB ultrotome equipped with LKB Cryokit. Dry ultrathin sections were mounted on grid meshes and were either air-dried, freeze-substituted or freeze-dried, and were covered with dry films of radioautographic emulsions, exposed, developed, stained and were observed in electron microscopes. After a number of trials, it was possible to obtain fairly good preservation of both cell structure and radioisotopes by means of freeze-dried and dry-mounted ultrathin frozen sections. However, the results are not completely satisfactory at the present time.

Animals↗