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Biomedical subjects

Y Tsuchihashi

Publications and source records attributed to Y Tsuchihashi.

70 records · Page 4Linked to original sources

Fluorescence fading and stabilization in cytofluorometry.

The factors affecting fluorescence fading in cytofluorometry were investigated using different kinds of nuclear staining, mounting media, and procedure of specimen preparation. Acceleration of fluorescence fading was observed in smear specimens treated with RNase, trypsin, or hypotonic solution before pararosaniline Feulgen nuclear staining. Similar effect was found for other DNA-stainings such as "33258 Hoechst" and Feulgen reactions with different Schiff-type dyes, such as acriflavine-SO2 and cresyl-violet-SO2, when chemically pure DNA was used. Fluorescence decay was rapid for all fluorochromes examined, when glycerin or buffer solution was used as mounting medium. Marked stabilization of fluorescence emission was induced in specimen mounted in non-fluorescent resin, Entellan (Merck), after post-staining fixation with absolute methanol for all tested fluorochromes. The same treatment induced almost complete fluorescence stabilization of fluorescein isothiocyanate (FITC); no detectable fluorescence fading was observed in a specimen stained with indirect immunofluorescence reaction using anti-UV-DNA antibody, during storage for 2 years at room temperature without special protection against light. These observations suggest that factors which bring about conformational stability of macromolecule-dye complexes generally induce fluorescence stabilization.

Animals↗

Comparison of cardiac lesions induced in rats by isoproterenol and by repeated stress of restraint and water immersion with special reference to etiology of cardiomyopathy.

Cardiac lesions induced in rats by isoproterenol, a potent beta-agonist, and by repeated stress of restraint and water immersion, in which sensitization of beta-adrenergic receptors would be expected to be induced, were investigated morphologically and following facts were revealed. 1) Cardiac lesions induced by isoproterenol, characteristic findings of which were myocardial hypertrophy, myocardial degeneration and myocardial necrosis replaced by interstitial fibrosis, were more analogous to cardiomyopathy than myocardial infarction or cardiac hypertrophy. 2) Cardiac lesions induced by repeated stress of restraint and water immersion, characteristic findings of which were myocardial hypertrophy, myocardial degeneration and myocardial necrosis replaced by interstitial fibrosis, were similar to those induced by isoproterenol. These results suggest that the endogenously induced dominant beta-adrenergic stimulating action during stress may play an important role in the pathogenesis of cardiomyopathy, the specific etiology of which is not yet known.

Adrenal Glands↗

Quantification of nuclear DNA and intracellular glycogen in a single cell by fluorescent double-staining.

It was found that intracellular glycogen is stabilized against acid treatment when it is stored under dry conditions for three months after methanol fixation. This stabilization allowed quantitative double fluorescence staining for nuclear DNA and intracellular glycogen, in a single cell. A Feulgen nucleal reaction, with acriflavine-Schiff's reagent following 5 N HCl hydrolysis at 25 degrees C for 4 min, was followed by a pararosanilin-Schiff PAS reaction for glycogen. This short term hydrolysis was found to be sufficient for the performance of a acriflavine-Schiff's Feulgen nucleal reaction and to provide good preservation of intracellular glycogen. Quantification of nuclear DNA and intracellular glycogen were consecutively carried out with a digital microfluorometer on a single ascites cancer cell of the AH-13 line stained by this method. It was found that there is a positive linear correlation between the amount of DNA and glycogen in this cell line.

Animals↗

Initial response of silver-impregnated "resting microglia" to stab wounding in rabbit hippocampus.

Adult rabbits received stab wound in the cerebrum and were sacrificed at intervals of 20, 30, and 39 h thereafter. Each animal was injected intracerebrally with 3H-thymidine 2h before fixation. Altered brain tissues of the stratum radiatum of hippocampus were taken for examination. Response of "resting microglia" to stab wounding was investigated by electron microscopic autoradiography and by autoradiography applied on silver-impregnated materials. Following results were obtained: (1) Resting microglia undergo marked swelling shortly after the brain damage. We designate these cells as "swollen microglia". (2) Swollen microglia form the only cell population that proliferate actively in the initial stage of glial response to the brain injury, and (3) astroglia do not proliferate during the same experimental periods, in the rabbit hippocampus.

Animals↗

Electron microscopic features of the resting microglia in the rabbit hippocampus, identified by silver carbonate staining.

Vibratome sections of hippocampus of adult rabbits were stained by a modified Hortega's silver-carbonate method. Impregnated materials were examined by electron microscopy to decide fine-structural characteristics of the resting microglia. Comparing their characteristics with those of macrophages, we came to the following conclusions: (1) Impregnated resting microglia in the hippocampus of adult rabbits can be identified as cells having distinct fine structures. (2) Resting microglia are morphologically different from macrophages or their precursor cells, and, therefore, seem not to be hematogenous cells sojourning in the normal brain parenchyma.

Animals↗

Telomerase activity and expression of telomerase RNA component and catalytic subunits in precancerous and cancerous colorectal lesions.

To investigate the role of telomerase activity in colorectal adenoma-carcinomas, telomerase activity, human telomerase RNA component (hTERC) and human telomerase reverse transcriptase (hTERT) mRNA were quantitatively analyzed in human cancerous and precancerous colorectal tissues. Sixty-six colorectal tumor specimens, including 10 invasive carcinomas, 6 mucosal carcinomas and 50 adenomas were evaluated. Ten specimens of normal tissue were also included in the study. Telomerase activity was assayed by semiquantitative fluorescence using the TRAP-eze(TM) telomerase detection kit. Analysis of the expression of each telomerase subunit gene was performed by real-time PCR amplification. There was a positive correlation between histological atypia and telomerase activity (rho = 0.700, p < 0.0001), hTERT mRNA expression (rho = 0.603, p < 0.0001), and hTERC expression (rho = 0.290, p < 0.05). There was also a positive correlation between the levels of hTERT mRNA and telomerase activity (r = 0.455, p < 0.01). Significant differences in the levels of hTERT mRNA were shown between normal tissues and the adenomas (p < 0.05) and between the mucosal carcinomas and invasive carcinomas (p < 0.05). The values of hTERC expression in neoplastic tissues were significantly higher than in the normal tissues; however, there were no significant differences between the adenomas and the carcinomas. In summary, although upregulation of hTERC expression is an early event in adenoma development, hTERT mRNA expression is gradually upregulated during the adenoma-carcinoma sequence and may be a rate-limiting determinant of telomerase activity.

Adenocarcinoma, Mucinous↗

Malignant endocrine carcinoma of the stomach.

Small cell carcinoma and small portions of poorly differentiated adenocarcinoma of the stomach sometimes differentiate into secretory cells. We experienced a pure endocrine carcinoma of the stomach. The patient, who had a giant mass in the greater curvature of the body of the stomach and regional lymph node metastasis, underwent total gastrectomy. Immunohistologic staining of the resected specimens revealed that the tumor was an endocrine carcinoma. The tumor cell shape in the present case was polygonal or oval and the cell size was relatively large. The cell nucleus was coarse chromatic and nucleoli were seen, whereas, the nucleus of small cell carcinoma often shows a fine chromatic pattern and nucleoli are not seen. On the basis of these findings, the present case was considered to be a pure endocrine carcinoma which was not classified as a small cell carcinoma.

Biomarkers, Tumor↗

Nerve contact with muscle component in neuromuscular hamartoma.

Neuromuscular hamartoma is a very rare soft tissue tumor, of which only 20 cases have been reported previously. None of these reports has described the relation between hamartomatous skeletal muscle and nerve fibers in the tumor. We experienced a patient with neuromuscular hamartoma arising at the brachial plexus. In this tumor, the localization of synaptophysin (SYP), S-100 protein (SP), neuron-specific enolase (NSE) neurofilament protein (NFP) and myoglobin (MG) was immunohistochemically detected. The results showed that SYP and MG were diffusely localized in the hamartomatous muscle fibers, SP in the schwann cells, and NSE and NFP in the axons of the hamartomatous nerve. Therefore, it is suggested that in the neuromuscular hamartoma, the structure of the neuromuscular junction may be similar to that in the motor end-plate of the normal muscle, but it may not be functional, because the hamartomatous muscles could not contract by nerve stimulation.

Hamartoma↗

Langerhans cell histiocytosis of an adult with tumors in liver and spleen.

We describe a 58-year-old male with multiple histiocytic tumors in the liver and spleen. Multiple tumors in the liver and spleen were seen by image analysis, and splenectomy showed a large splenic tumor with a small nodule and a swelling lymph node in the hilus. Histological features of the tumors in the liver and spleen revealed proliferation of histiocytic cells with large and clear cytoplasm and a horseshoe-shaped nucleus. Immunohistochemical studies revealed the presence of S-100 protein and CD1a antigen in the tumor cells, and neither lymphocytic marker nor lysozyme was detected. No definite Birbeck granules were seen ultrastructurally, thus the tumor cells could be classified into Langerhans cell type without Birbeck granules. Administration of adriamycin, vincristine, cyclophosphamide and prednisolone reduced size and number of the liver tumors, and the histiocytic cells could not be detected in repeatedly biopsied tissue from liver tumor. We present the clinical, immunohistological and cytological features in a visceral type of adult Langerhans cell histiocytosis, which responded well to chemotherapy.

Antigens, CD1↗