著者

依田 敏行 後藤 由夫

出版者

一般社団法人 日本臨床化学会

雑誌

臨床化学シンポジウム (ISSN:03863417)

巻号頁・発行日

vol.21, pp.20-22, 1982-07-15 (Released:2012-11-27)

参考文献数

7

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The detecting ability of the photon counting system of single photoelectron counting method has been greatly improved and an equipment of this method, designed for biochemical and biomedical applications, was developed. The photomultiplier used in the instrument was of bialkali photocathode of 5cm diameter with the spectral responce nearly from 300nm to 660nm and of specifically low noise selected from hundreds of the same photomultipliers. The photomultiplier was cooled down to -20° in the equipment in the actual photon counting for the further reduction of the noise. All other possible electrical noises from the inside and the outside of the counting machine were eliminated or minimized. The sensitivity of the photon counting equipment reached the level high enough to permit the determination of extremely weak light emission like those of human blood and tissue samples. Thus, the first quantitative measurement, to our knowledge, of ultra weak chemiluminescence of human blood samples was performed with this instrument. The whole blood and plasma samples of normal subjects gave relatively low levels of light emission. On the contrary, the blood samples of patients with diabetes mellitus and with liver diseases showed significantly higher light emission levels. Of particular interest was of the blood of patients with obstructive jaundice and of severe hepatitis. The light emission of the blood samples of these disease was generally very high and in the levels allowing the spectral analysis of the emitted light.The analysis and the scavenger experiments indicated the contribution of singlet oxygen as the photon emitting entity in these blood samples.

著者

飯田 久也 川出 眞坂 岡野 幸雄 野沢 義則

出版者

一般社団法人 日本臨床化学会

雑誌

臨床化学シンポジウム (ISSN:03863417)

巻号頁・発行日

vol.20, pp.15-20, 1981-07-15 (Released:2012-11-27)

参考文献数

5

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The relationship between morphology and lipid composition of red blood cells in patients with hepatobiliary disease has been clearly demonstrated by several investigators. Little information is, however, available regarding the mechanism by which erythrocyte membrane abnormalities are induced. In order to clarify such mechanism, biochemical, ultrastructural and physiochemical studies of red blood cell membranes from patients with biliary obstruction and congenital biliary atresia have been carried out.Scanning electron microscope observation revealed that red blood cells from patients with biliary atresia classified as Group I (red cell lecithin≥41%) are mostly of the “target” type. On the other hand, red blood cells from Group III (red cell lecithin<31%) are almost normal biconcave disks.Phospholipids and free cholesterol were increased strikingly in red blood cells from Group I, whereas no significant difference was observed between Group III and control. The abnormalities in the lipid profile as well as morphology are dependent upon the degree of biliary obstruction. It is obvious that increase in the phospholipid content is principally due to an increment of lecithin. The analysis of the fatty acid composition of lecithin revealed a similar pattern in red blood cell membranes, plasma and bile. It is of particular interest that the lecithin fatty acid composition of lipoprotein-X (LP-X) was found to be quite similar to that of erythrocyte membranes. On the other hand, in spite of little change in the lipid content of erythrocyte membranes, the lecithin fatty acid composition of erythrocyte membranes and plasma in Group III showed a similar but smaller change compared to that of patients in Group I.Freeze-fracture electron microscopy showed significant alterations in Group I membranes, such as depressions on the fracture faces and reduced density of membrane particles (3,584→2,432/μm2) on the protoplasmic face.Electron spin resonance studies on erythrocyte membranes labeled with 5-nitroxide stearate demonstrated that the erythrocyte membranes from Group I are more fluid than the control membranes.Consequently, two working hypotheses were proposed by which excess membrane may be incorporated into erythrocyte membranes; fusion of LP-X with erythrocyte membranes (Group I) and exchange of lipids between erythrocyte membranes and plasma lipoproteins (Group III).Red blood cells from patients with congenital biliary atresia exhibited a wide variation in the morphology, such as target, spur and cup formed cells. By freeze-fracture electron microscopy, membrane particle-free areas were prominent in the fracture faces and the density of membrane particles were 20% lower than those of control subjects.Both free cholesterol and phospholipids were found to be increased in erythrocyte membranes from patients with congenital biliary atresia. In addition, the fatty acid composition of erythrocyte lecithin was similar to that of acquired biliary obstruction.Fluorescence polarization studies by using fluorescent probe, 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) on red blood cells demonstrated that erythrocyte membrane lipids from patient are less fluid than those of control subject.From the similarity of lipid alterations to those observed with acquired biliary obstruction and the fact that LP-X was found in all patients examined, it is reasonable to consider that LP-X might be involved in causing erythrocyte membrane abnormalities in congenital biliary atresia.

著者

北村 元仕 仁科 甫啓 飯島 直子 橋本 史子 三輪 史朗 掛橋 順子

出版者

Japan Society of Clinical Chemistry

雑誌

臨床化学シンポジウム (ISSN:03863417)

巻号頁・発行日

vol.11, pp.178-181, 1972

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A case of complete deficiency of lactate dehydrogenase subunit H has been presented. Deficiency of LDH H-subunit of this case is one of inborn errors of metabolism. The propositus (J. A.) is a 64 year-old male with mild diabetes, which has been under good control by restricted diet and torazamide. Low serum LDH activity was noticed upon recent laboratory check-up, and finally, complete absence of LDH H-subunit was discovered by serum isoenzyme study. In addition, the erythrocytes, leukocytes, platelets as well as saliva of the propositus were found to be deficient in LDH H-subunit. LDH of the propositus was composed only of M-subunit. Hence, the LDH isoenzyme patterns of the serum, blood cell and saliva in the propositus showed only one band, LDH-5 (MMMM tetramer). It is of special interest that erythrocytes fructose 1, 6-diphosphate, dihydroxyacetone phosphate, as well as glyceraldehyde 3-phosphate concentration in the propositus were found to be definitely increased. These data strongly suggest that disturbance in erythrocyte glycolysis exists at the glyceraldehyde 3-phosphate dehydrogenase step. However, the erythrocyte glyceraldehyde 3-phosphate dehydrogenase activity in this case was within the normal range, which suggests indirectly that the cause of this disturbance in glycolysis might be due to high NADH and relatively low NAD caused by block of LDH step. In the whole blood of the propositus, lactate concentration came out to be low, whereas that of pyruvate was within the normal range.<BR>The studies on reduction of methemoglobin with glucose and lactate suggest that LDH activity plays only a minor role in the reduction of methemoglobin in human erythrocytes.