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Volume 9 (1976)
- Issue 4 Pages 269-
- Issue 3 Pages 197-
- Issue 2 Pages 107-
- Issue 1 Pages 1-
Volume 9, Issue 2
Displaying 1-8 of 8 articles from this issue
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NAGAYASU OTSUKA, TOSHIHIKO KISHIMOTO, TAKASHI NAGITA1976 Volume 9 Issue 2 Pages 107-110
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSThe hippocampal formation of monkey was studied by sulfide silver technique. Intense positive reaction to this method was found in the stratum radiatum of h3, h4 and h5 of the hippocampus, as well as the stratum multiforme of the fascia dentata. All these areas correspond to the endings of mossy fibers at the apical dendrite of pyramidal cells in the hippocampal formation.View full abstractDownload PDF (2913K) -
YOSHIHO KATAYAMA, HIROSHI KOBATAKE, HANZABURO SHIDA1976 Volume 9 Issue 2 Pages 111-124
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSA new histochemical method for β-hexosaminidase is presented as a modification of Hayashi's technique.
N-acetyl-β-D-hexosaminides of Naphthol AS-BI were prepared by treating the anisidide with acetochlorohexosamines in acetone with an aqueous alkali. These compounds were recrystallized several times and used as the substrate.
The pH of the medium could be desirably reduced near to the optimal pH. In simultaneous histochemical techniques, some diazonium salts were tested in order to determine their resistance to diffusion or their coupling rates. Fast red violet LB salt was more satisfactory as a coupling dye. No dye depositions were observed in the cells of various tissue samples above 50μM, of N-acetyl-glucosaminolactone and N-acetyl-galactosaminolactone.
Frozen sections of the tissues were incubated at 37°C together with substrates in the presence of fast red violet LB salt at pH 4.4. Many fine granules associated with enzyme activity were found to be localized uniformly throughout the cytoplasm except for the nucleus. For the N-acetyl-β-glucosaminidase activity, the marked dye deposits were observed in some organs, such as epididymis and kidney. Dye deposits in the nerve cells of the brain and the spinal cord were also observed. For the N-acetyl-β-galactosaminidase activity, moderate dye deposits were found in rat tissue specimens. The latter enzyme, however, showed a tendency to be slightly less active than N-acetyl-β-glucosaminidase.View full abstractDownload PDF (4523K) -
YOSHINOBU HOSHINO, W. ALLEN SHANNON Jr., ARNOLD M. SELIGMAN1976 Volume 9 Issue 2 Pages 125-136
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSUltrastructural and limited ultracytochemical studies were explored with potassium ferrocyanide-reduced osmium tetroxide (POCC). The use of 2% osmium tetroxide solution following POCC treatment was found to improve the preparation of ultrathin sections by facilitating the infiltration of Epon into the treated tissues. Potassium ferrocyanide alone did not stain tissues. Treating of tissues with potassium ferrocyanide and osmium tetroxide separately was not effective indicating that reduced osmium staining could not be produced within the tissues.View full abstractDownload PDF (23894K) -
MASAO OHYUMI, SADAMU TAKANO1976 Volume 9 Issue 2 Pages 137-143
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSInosine-5-phosphate dehydrogenase was histochemically demonstrated in cryostat sections of the human esophagus cancer. Suitable conditions for histochemical reaction of the enzyme activity in this tissues were determined. The incubation mixture containing 50mg inosine-5-phosphoric acid, 10mg nicotinamide adenine dinucleotide, 10mg Nitro-BT (1mg/ml), 10mg reduced glutathione, 2mg KCl, 10ml Tris-HCl buffer (0.2M pH 7.1) was proposed, while control procedures was prepared, using the incubation mixture without inosine-5-phosphate.View full abstractDownload PDF (2926K) -
KOSUKE CHIDA, NOBORU YAMAMOTO, KENJIRO YASUDA1976 Volume 9 Issue 2 Pages 144-155
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSFrom rat liver has been purified sorbitol dehydrogenase in high purity, showing a single line by means of disc electrophoresis. The substrate specificity of the purified enzyme was very low; the Km value of the enzyme was 4.5×10-4M for sorbitol, 2.6×10-4M for xylitol and 2.4×10-3M for ribitol, respectively. The localization of sorbitol dehydrogenase was investigated by use of both fluorescein-labeled and peroxidase-labeled antibody methods on the light microscopic level. The antigen was recognized concentratedly around the nucleus and in less amount in the other parts of the cytoplasm of the liver cells in a granular profile. In the electron microscopic study by means of peroxidase-labeled antibody technique, the positive deposits were observed in the reticulated or net-like pattern in the amorphous part of the cytoplasm of parenchymal cells, which might correspond to the glycogen area. The results of the present study generally coincided with those of the foregoing report, in which the proper enzyme was demonstrated by using a histochemical technique in light and electron microscopy.View full abstractDownload PDF (8815K) -
AKIO SUMITANI, JUZO SAWANO, HIROIKU KAWAKAMI, AKIMA MIYOSHI, AKIRA IKE ...1976 Volume 9 Issue 2 Pages 156-167
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSThe fluorescent antibody method is widely used as a routine procedure for the histochemical examination of enzymes. In the present paper, the antiserum used in such procedures was applied to quantitative immunoelectrophoresis for the microquantification of enzymes. Bovine liver tissue in amounts corresponding to that available from liver biopsies was used to perform quantitative determinations of arginase which is the key enzyme in the urea cycle.
Antiserum was obtained by sensitization of a rabbit using standard crystalline bovine liver arginase, which had been obtained from the Boehlinger Mannheim Company and purified by disc electrophoresis. This antiserum was mixed with 1% agarose, the supporting medium, and then quantitative immunoelectrophoresis was performed using 5μl of the supernatant of bovine liver homogenate.
The area enclosed within the precipitation lines that had been produced as a result of electrophoresis was confirmed to be proportional to the enzymatic activity. Furthermore, based upon the knowledge that the standard arginase had an enzymatic activity of 70 units per mg, the enzymatic activity units were calculated for the area enclosed within the precipitation lines.
It was thus demonstrated that microquantification is possible. In addition, the DNA content within the precipitate that had been produced during the preparation of the sample solution was measured and used as the index of the cell count. This value was used to calculate the enzymatic activity per mg DNA.View full abstractDownload PDF (10941K) -
MASAOKI HARADA, KAZUHIKO OKABE, KAZUTOSHI SHIBATA, HIROTAKE MASUDA, KE ...1976 Volume 9 Issue 2 Pages 168-179
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSHistochemical demonstration of γ-glutamyl transpeptidase (γ-GTP) during chemically induced hepatocarcinogenesis in the rats was studied. The carcinogens employed were N-OH-FAA (N-hydroxy derivative of N-2-fluorenylacetamide), and aflatoxin B1.
In the control rats, activity of γ-GTP was demonstrated in the bile duct cells and occasionally along the bile canaliculi, but was not in the hepatocytes. Proliferating bile duct cells including so-called oval cells and cholangiocarcinoma cells, and hepatocytes of hyperplastic areas and nodules and hepatocellular carcinoma induced by N-OH-FAA with or without aflatoxin B1 revealed positive activity of γ-GTP, histochemically. Only focal demonstration of γ-GTP activity in the hepatocyte was observed in association with a mild hyperplastic change in rats treated with aflatoxin B1 alone.
The elevation of γ-GTP activity in serum and liver tissue with appearance of fetal type of its isoenzyme pattern was also demonstrated biochemically and correlated with the histochemical appearance of γ-GTP activity in the liver tissue from the early phase of hepatobiliary carcinogenesis.
These results seem to demonstrate carcino-embryonic nature of the induced γ-GTP in the development of cholangiocellular and hepatocellular carcinomas.View full abstractDownload PDF (6042K) -
MASARU FUKUDA, KAZUO NAKANISHI, TERUO MUKAINAKA, AKIHIRO SHIMA, SETSUY ...1976 Volume 9 Issue 2 Pages 180-192
Published: 1976
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSImmunofluorescent staining for nuclear UV photoproducts was combined with nuclear Feulgen reaction for cytofluorometry to quantitate the potency of excision repair of the lesions per unit amount of DNA on a single cell,
Isolated hepatocytes and cerebellar neurons of adult rat were UV-irradiated and made react with rabbit antiserum containing antibodies to photoproducts of DNA after UV-irradiation, The UV induced lesions were demonstrated by indirect immunofluorescent technique using FITC labeled anti-rabbit's immunoglobulin antibody. The immunofluorescent staining was combined with subsequent Feulgen nuclear reaction after stabilizing the fluorescent dye-immunoglubulin complex by post-fixation with absolute ethanol.
As a linear relationship was found between fluorescence intensity and concentration of free and protein-bound FITC up to absorbance value of 0.1, intensity of the immunofluorescent staining was controled not to exceed this limit.
Two peaks of fluorescence of FITC (520nm) and Feulgen dye (613nm) were separately identified on a double-stained nucleus with respective excitation by 450nm and 543nm without interference of each other fluorescence, Proportionality between DNA content and amount of Feulgen fluorescence was preserved in the double stained hepatocytes and cerebellar neurons, Linear relationships were noted between the amounts of Feulgen DNA and nuclear FITC in the same cells immediately after UV-irradiation. The results indicate that the amounts of photoproducts produced in unit amount of nuclear DNA are equal in all cells irrespective of the kinds of tissues and stages of cell cycle.View full abstractDownload PDF (6671K)
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