The Semi-Quantitative Isolation of Phosphatidyl-Serine From Pig Erythrocytes, Using Cellulose Column Chromatography.

1. By cellulose column chromatography, phosphatidyl-serine was isolated from three fractions, obtained from pig erythrocyte lipids by silicic acid column chromatography. The total yield was 81% on the basis of the value determined by Dawson\u27s method. 2. One of the preparations (yield : 27%) was confirmed to be in high purity, by elemental analyses, by infrared spectroscopy and by various paper- and thin layer-chromatography. 3. The occurrence of phosphatidyl-serine in two different ionic forms ; mono-basic and di-basic ones, is shown by cation analyses as well as by infrared spectra. The cations in the native preparations are composed mainly of bivalent ions such as Ca++ and Mg++. 4. The gas chromatographic data showed that the fatty acids of phosphatidyl-serine were composed mainly of C-18 acids (91.7%) ; stearic (31.4%), oleic (54.5%), and linoleic acid (5.2%)and of C-16 acid ; palmitic acid (5.3%)

Studies on Pig Erythrocyte Lipids (1). Fractionation of Erythrocyte Lipids by the Simple Combination of Silicic Acid and Cellulose Column chromatography.

1. The fractionation of pig erythrocytes lipids into the eight fractions, termed as S-0, S-2, S-5, S-10, S-20, S-25, S-40 and S-50 respectively, was carried out by silicic acid column chromatography, with determination of phosphorus and sugar contents in each fraction. The identification of lipids in each fraction was performed also by thin layer and silicic acid impregnated paper chromatography as well as infrared spectroscopy. 2. The chromatographic data showed that the fraction S-0 was composed mainly of cholesterol with a minor quantity of cholesterolester and/or monoglyceride, S-5 mainly of fatty acid, S-10 of cerebroside, cardiolipin and/or phosphatidic acid, S-20 of phosphatidyl-ethanolamine and phosphatidyl-serine, S-25 of phosphatidyl-ethanolamine and globoside, S-40 of phosphatidyl-choline and phosphatidyl-serine and S-50 of sphingomyelin with a minor quantity of phosphatidyl-serine and phosphatidyl-choline. 3. The last four fractions such as S-20, S-25, S-40 and S-50 were further separeted into four fractions (C-0, C-10, C-20 and C-50, respectively) by cellulose column chromatography with the determination of their phosphorus and sugar contents. The identification of the lipids in each fraction was performed by thin layer chromatography.4. The chromatographic data indicated that all the fractions C-0 derived from S-20, S-40 and S-50 were composed mainly of phosphatidyl-serine, while the fraction C-0 from S-25 mainly of globoside ; the fractions C-10 mainly of non-acidic phospholipids, that is, the fraction S-20 C-10 of phosphatidyl-ethanolamine, S-40 C-10 of phosphatidyl-choline, and S-50 C-10 of sphingomyelin ; the fraction C-50 mainly of globoside. 5. It was noted that phosphotidyl-serine wes eluted over wide range of fractions by silicic acid column chromatography, which was shown to be unsuitable for removal of phosphatidyl-serine from the common phospholipids, although phosphatidyl-serine could be well separated from the non-acdic phospholipids by cellulose column chromatography. The latter method was also utilizable for the separation of globoside from the other lipids. 6. The final identification of individual lipids will be presented in detail in the succeeding papers

Studies on Pig Erythrocyte Lipids (II). The Isolation and the Identification of Individual Lipids with Some Quantitative Aspects on the Composition of Erythrocyte Lipids.

1. The individual lipids in pig erythrocytes were isolated from the fractions obtained by the method simply combined silicic acid- and cellulose-column chromatography reported in the preceding paper. 2. Respective identifications of the less-polar lipid preparations as cholesterol-ester, triglyceride, free cholesterol and free fatty acids were performed by their chromatographic studies and by infrared spectroscopy. 3. The major phospholipids, phosphatidyl-ethanolamine, phosphatidyl-choline, phosphatidyl-serine and sphingomyelin were isolated in the high yields, and corresponded to 80-97% of the value determined by Dawson\u27s method. 4.The identification of the isolated phospholipids and sphingolipids, that is, cardiolipin, phosphatidyl-ethanolamine, phosphatidyl-choline, sphingomyelin, ceramide, ceramide-mono-glucoside and globoside in which the data of phosphatidyl-serine only was presented in the preceding paper, was performed by chemical analysis, infrared spectroscopy and thin layer- as well as silicic acid impregnated-paper chromatography. 5. The fatty acid of various lipids, such as free fatty acid, ceramide,ceramide-mono-glucoside, cardiolipin, phosphatidyl-ethanolamine, phosphatidyl-serine, phosphatidyl-choline, sphingomyelin and globoside was analysed by gas-chromatography. Some considerations regarding the fatty acid compositions were also presented. 6. The long chain base and its derivatives, which were isolated from the sphingomyelin preparation by alkaline methanolysis as well as from globoside by acid methanolysis after the deacylation by alkaline treatment, were identified as erythro-sphingosine. It was also noted that the long chain base obtained from sphingomyelin by alkaline methanolysis gave only one spot identical with erythro-sphingosine on the thin layer chromatogram but the base from ceramide separated into two spots, the upper one iden-tical with erythro-form and the other lower one identical with threo-form. The results suggest that there could be two kinds of ceramide such as erythro- and threo-forms in pig erythrocytes

A new phenotype of mitochondrial disease characterized by familial late-onset predominant axial myopathy and encephalopathy

Axial myopathy is a rare neuromuscular disease that is characterized by paraspinal muscle atrophy and abnormal posture, most notably camptocormia (also known as bent spine). The genetic cause of familial axial myopathy is unknown. Described here are the clinical features and cause of late-onset predominant axial myopathy and encephalopathy. A 73-year-old woman presented with a 10-year history of severe paraspinal muscle atrophy and cerebellar ataxia. Her 84-year-old sister also developed late-onset paraspinal muscle atrophy and generalized seizures with encephalopathy. Computed tomography showed severe atrophy and fatty degeneration of their paraspinal muscles. Their mother and maternal aunt also developed bent spines. The existence of many ragged-red fibers and cytochrome c oxidase-negative fibers in the biceps brachii muscle of the proband indicated a mitochondrial abnormality. No significant abnormalities were observed in the respiratory chain enzyme activities; however, the activities of complexes I and IV were relatively low compared with the activities of other complexes. Sequence analysis of the mitochondrial DNA from the muscle revealed a novel heteroplasmic mutation (m.602C>T) in the mitochondrial tRNAPhe gene. This familial case of late-onset predominant axial myopathy and encephalopathy may represent a new clinical phenotype of a mitochondrial disease

MAGNETIC RESONANCE IMAGING AND PATHOLOGIC STUDIES ON LATERAL FLUID PERCUSSION INJURY AS A MODEL OF FOCAL BRAIN INJURY IN RATS

In this study, morphologic changes in brain lesions initiated by moderate lateral fluid percussion injury in rats were investigated chronologically using high resolution magnetic resonance imaging (MRI) and histopathologic methods. Rats were subjected to moderate fluid percussion injury (average 2.80±0.48 atmospheres) over the exposed dura overlying the right parietal cortex. MRI obtained in vivo were compared with corresponding pathologic findings at 1, 6, and 24 h and at 3, 6,14 and 80 days after injury. T2 weighted images showed scattered low signal intensity in the injured cortex within a few hours after injury, whereas histologic findings revealed intraparenchymal hemorrhages. T2-weighted images of the ipsilateral cerebral cortex and/or corpus callosum showed a high-signal-intensity area 4 h after injury. The high-signal-intensity area became largest in size between 6 and 24 h, then declined gradually, and almost disappeared 14 days after injury. Histologic examination revealed pyknosis, retraction of the cell body of neurons with vacuolated neuropil in the corresponding regions 6 and 24 h after injury, and cystic necrosis 14 days after injury. The location and extent of these pathologic changes were depicted accurately by MRI in vivo. In the hippocampus, pyknosis and retraction of the cell body of pyramidal neurons were observed on the injured side 24 h after injury, and the number of neurons in the CA1 and CA2『 CA3 regions decreased significantly on the same side by 14 days after injury. It is concluded that morphologic changes in the brain following experimental traumatic brain injury in rats are detectable in υivo by high-resolution MRI, and that MRI may be useful for the evaluation of treatment effects in experimental brain injury

Stndies on the Pneumoconiosis of Electric Welders

Sadatoshi, one of the authors, had reported previously that he had found 26% of electric welders at a certain heavy industrial plant suffered from pneumoconiosis by roentgenological examination. In this time investigation was done upon 78 electric welders carrying pneumoconiosis without pulmonary tuberculosis and the result was as follows: 1) Chest X-ray findings approved granulated type. No fibrosis, tendency of confluence, emphysema and eggshell calcification. Changes in hilus could not be found at the begining. 2) No visible changes in blood findings. Except, as the type advanced, a tendency of a slight decrease in neutrophil leukocytes and increase in lymphocytes could be seen. 3) No remarkable changes admitted in lung vital capacity and maximum ventulating volume. Expiratory impediment appeared in the third type. 4) Though welding dust contains SiO(2), welder's pneumoconiosis is innocent compared with silicosis. It also seems a little different from siderosis and so is suitable to classify it as "welder's Pneumoconiosis". 5) Occupational career should be cared when taking routine X-rayphotographs