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Biomedical subjects

T Miyatake

Publications and source records attributed to T Miyatake.

At least 325 records · Page 18Linked to original sources
PubMed1983

Isolation and characterization of the trisialogangliosides from bovine adrenal medulla.

Trisialogangliosides were isolated from bovine adrenal medulla by DEAE-Sephadex A-25 and Iatrobeads column chromatography. Their structures were elucidated by sugar analysis, neuraminidase digestion, and permethylation studies. The complete structures of trisialogangliosides, A to D, were identified as follows. A: GT1b, IV3NeuAc, II3 (NeuAc)2-GgOse4Cer. B: GT1b(NeuAc/NeuAc-NeuGc-); IV3NeuAc, II3 (NeuAc alpha 2-8 NeuGc-)GgOse4Cer. C: GT1b (NeuGc/NeuAc-NeuAc-); IV3NeuGc, II3 (NeuAc alpha 2-8 NeuAc-)GgOse4Cer. D: GT1b (NeuAc/NeuGc-NeuGc-); IV3NeuAc, II3 (NeuGc alpha 2-8 NeuGc-)GgOse4Cer. Gangliosides B, C, and D, which contain N-glycolylneuraminic acid, have not previously been reported in the literature.

Adrenal Medulla↗
PubMed1983

[Electrophoresis of proteins of muscle fibers with rimmed vacuoles].

This is the first report on the analysis of the proteins in the single muscle fibers with rimmed vacuoles. A new method was developed to analyse single muscle fibers which had been histologically characterized in the freeze-dried sections of muscles. Muscle biopsy was performed in three patients with distal myopathy and one patient with Kearns-Sayre-like syndrome associated with rimmed vacuoles (Table 1). Ten (30 micron thick) frozen serial sections were dried according to Lowry and Passonneau, and further five consecutive sections were cut and stained with usual histochemical methods including Gomori trichrome and NADH-TR staining. Each muscle fiber in the freeze-dried sections could be identified with the corresponding one in the stained sections, and it was possible to determine whether the fiber had rimmed vacuoles or not (Fig. 2). Particular fibers were identified in the consecutive freeze-dried sections and cut out from three sections with no contamination of the interstitial tissue, and they were subjected to SDS gel electrophoresis. Resultant gel patterns showed substantial increase of 55 k dalton band, the extent of which was much more larger in that of the fiber with rimmed vacuoles than that without vacuoles (Fig. 3). This feature was common to all of the gels from four patients. A portion containing a number of muscle fibers of a freeze-dried section from each of the four patients and the two controls was subjected to two-dimensional gel electrophoresis (2-D). Of 55 k dalton proteins, three spots around the isoelectric point of alpha-actin were found to be increased (Fig. 4).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1983

[31P-NMR analysis of high energy phosphorous compounds (ATP and phosphocreatine) in the living rat brain--effects of halothane anesthesia and a hypoxic condition].

31phosphorus nuclear magnetic resonance (31P-NMR) measurements have provided new and valuable insights for studying the metabolism of living systems. The aim of this paper is to introduce a technique of application of 31P-NMR measurements using a surface coil method, and to discuss the effects of halothane anesthesia and hypoxic hypoxia on the energetic metabolism of intact rat brains. All measurements were made using a JEOL FX 270 spectrometer with a super conducting magnet of 54-mm bore diameter. The magnetic field intensity of this machine is 6.3 tesla, and the resonance frequency used for 31P was 109.14 MHz. We remodelled an ordinary probe to take a live rat, and the animals were made to inhale anesthetic halothane or mixture of oxygen and nitrogen at various concentrations controlled by a flow regulator. The best conditions for measurements with our surface coil method were determined in this study as follows: (1) 90 degrees pulse width and selectivity, Fig. 1 shows signal selectivity in depthwise direction changed with 90 degrees pulse width, which was set to 20 microseconds. (2) Sensitivity and resolution; To obtain a spectrum of 31P-NMR from a rat brain 500 accumulations of free induction decays were considered suitable for both time and space resolution. Fig. 2 shows variations of signal intensity with pulse repetition time, which was set to 2 sec. It took about 17 min for averaging to get a spectrogram. (3) Quantitative accuracy and qualification; As shown in Fig. 3, a linear relationship was found between the signal intensity of beta-phosphate of ATP and the concentration of ATP solutions, thus proving the quantitative accuracy of our systems.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1982

Disialogangliosides in bovine adrenal medulla.

Gangliosides were isolated from bovine adrenal medulla by DEAE-Sephadex A-25 and Iatrobeads column chromatography. The disialo fraction contained five major gangliosides, A to E; together they accounted for 4.6% of the total lipid-bound sialic acid. On the basis of sugar analysis, permethylation, mild acid hydrolysis and neuraminidase degradation studies, the structures of these five gangliosides are characterized as follows: (formula: see text). The major fatty acids of the disialogangliosides were C16:0, C18:0, C22:0, C23:0, C24:0, and C24:1, which were characteristic of extraneural gangliosides.

Adrenal Medulla↗
PubMed1982

Radioassay method of neuraminidase towards N-acetylneuraminosyl hexasaccharides.

We devised a sensitive method to assay for neuraminidase activities towards alpha-(2 leads to 3)-N-acetylneuraminosyl hexasaccharide and alpha-(2 leads to 6)-N-acetylneuraminosyl hexasaccharide, which were isolated from the urine of a patient with adult sialidosis with partial deficiency of beta-galactosidase. Standard assay conditions for the determination of these neuraminidase activities were established and the radiolabeled reduced derivatives of these substrates were used. The fibroblast neuraminidase had its maximum activity at pH 4.0-4.2, with Km values of 2.22 X 10(-3) and 4.17 X 10(-3) mol/l and Vmax values of 76.9 and 28.6 nmol . mg-1 protein . h-1 towards the 2 leads to 3 isomer and the 2 leads to 6 isomer, respectively. Neuraminidase deficiencies were found in the fibroblasts of adult sialidosis, mucolipidosis II and III. These studies were compared with the neuraminidase activity towards alpha-(2 leads to 3)-N-acetylneuraminosyl lactose.

Carbohydrate Metabolism, Inborn Errors↗
PubMed1982

Neuraminidase deficiency and accumulation of sialic acid in lymphocytes in adult type sialidosis with partial beta-galactosidase deficiency.

Neuraminidase deficiency has been demonstrated in cultured skin fibroblasts of patients who have adult type sialidosis with partial beta-galactosidase deficiency. A substantial amount of residual enzyme activity has been observed in leukocytes, however. To explain this discrepancy, the nature and distribution of the enzyme were studied. Neuraminidase activity was higher in lymphocytes than in granulocytes of normal controls. In patients' lymphocytes, neuraminidase activity was profoundly decreased and total sialic acid contents were increased 2.3-fold. Two neuraminidases, one sonication-labile and the other sonication-stable, were found in lymphocytes; the former was predominant in cultured skin fibroblasts. The defective enzyme in this disorder was found to be the sonication-labile neuraminidase in both cultured skin fibroblasts and lymphocytes.

Cerebellar Ataxia↗
PubMed1982

Neuraminidase activities in sialidosis and mucolipidosis.

Defects of neuraminidase activities towards sialyloligosaccharides in fibroblasts and leucocytes and enhanced excretion of sialyloligosaccharides in urine were shown in patients with adult type sialidosis with partial deficiency of beta-galactosidase and cherry red spot-myoclonus syndrome. No differences in their neuraminidase residual levels and urinary excretion patterns on thin-layer chromatography were found between these two disorders. In mucolipidosis II and III patients, the neuraminidase activities towards sialyloligosaccharides were almost normal in leucocytes, although decreased in fibroblasts. The discrepancy of neuraminidase activities towards 2 leads to 3 and 2 leads to 6 sialyloligosaccharide isomers was not noticed in all cases.

Chromatography, Thin Layer↗
PubMed1982

Eicosapolyenoic acids of serum lipids of Japanese islanders with low incidence of cardiovascular diseases.

Japanese are unique among the peoples of developed countries in having a high intake of eicosapentaenoic acid (C 20:5) from fresh fish and this may in part contribute to their low incidence of cardiovascular diseases. Mass spectroscopic analyses of eicosapolyenoic acids (C20:3, C20:4 and C20:5) were carried out on the serum of aged persons living on Kohama island in Okinawa and known to have the lowest incidence of cardiovascular diseases in Japan. All but 4 of the 77 persons examined (73.94 +/- 7.81 years old) led active fishing-farming lives. The total amount of eicosapolyenoic acids in the serum of persons on Kohama island (46.77 +/- 7.46 mg/100 ml) was higher (p less than 0.001) than that in people on mainland Japan, owing to the higher intake of fresh fish (147.7 g/day). A positive correlation (p less than 0.01) was found between serum C 20:5 concentration (6.82 +/- 2.54 mg/100 ml) and high density lipoprotein concentration (55.38 +/- 13.83 mg/100 ml). In addition, there were positive correlations (p less than 0.01) between serum C 20:3 concentration (6.58 +/- 1.61 mg/100 ml) and total cholesterol (188.60 +/- 32.30 mg/100 ml), and triglyceride and skinfold thickness. The blood pressure level (p less than 0.01), incidence of abnormal ECG (p less than 0.05), and salt intake (6.2-8.3 g/day) estimated from urinalysis, were all lower than the average figures for Japanese of similar ages. No persons examined showed Q-wave on ECG. The percentage of smokers and drinkers were similar for Kohama island and mainland Japan.

Activities of Daily Living↗
PubMed1982

Measurement of eicosapolyenoic acids in the serum by gas-liquid chromatography-chemical ionization mass spectrometry.

The use of octadeutero eicosatetraenoic acid as an internal standard for the reproducible measurement of serum eicosapolyenoic acids (C20:3, C20:4, and C20:5) by gas-liquid chromatography-chemical ionization mass spectrometry is described. The method has the following advantages. The physicochemical properties of the internal standard and the eicosapolyenoic acids are similar. The acids are easily separated from compounds of similar retention times by means of selected ion monitoring. The measurements can be made more rapidly, (10 min per sample) than with previous techniques.

8,11,14-Eicosatrienoic Acid↗
PubMed1982

Characterization of GM1 ganglioside by direct inlet chemical ionization mass spectrometry.

Intact permethylated and permethylated-reduced (LiAlH4) derivatives of GM1 ganglioside were analyzed by direct inlet ammonia chemical ionization (CI) mass spectrometry. In addition, the trimethylsilylated derivative of the permethylated-reduced sample of this ganglioside was similarly analyzed. CI mass spectrometry proves to be a highly satisfactory method for structural studies of GM1 ganglioside because of the simple fragmentation pattern and the presence of prominent molecular ions and fragment ions in the high mass region. Complete information on the carbohydrate sequence and lipophilic composition can be easily obtained.

Animals↗