Acta Medica Okayama volume23 issue5

1. The cells used in the present experiments were lymph-node cells from inbred mice, and over 98 % cells were proven to be small lympho-cytes. Therefore, those cells that have undergone blastformation are all those derived from small lymphocytes. 2. When homogenate of one cell group is cultured with live cells of the other pairing group, there occurs blastformation. In the presence of PHA, such a blastformation becomes more marked. 3. The optimal concentration of PHA (phytohemagglutinin)-M added to the mixed culture is found to be 1% (v/v). 4. The maximum rate of blastformation in the mixed culture is observed at the culture hour 48, being much faster than in the mixed culture between two live cell groups. 5. In the mixed cultures between subcellular fractions prepared from cell homogenate by centrifugation and live cells, the transplantation antigenic potential (histocompatibility antigenic potential) is seen in the mitochondrial and the microsomal fractions, especially marked in the latter. 6. In the observations carried out by various combinations of these inbred mice, it has been demonstrated that the rate of blastformation induced by the addition of cell homogenate or sediment fractions prepared from the homogenate reflects quite accurately the differences in H-2 antigens.

The author has described modern thrombolytic therapy of arterial and venous thrombosis and emboli by therapeutic fibrinolysis and other drugs also methods and effects of local and parenteral application of fibrinolysin preparations, dosage, control, indications. Contraindications, side-effects and their treatment with fibrinolysin antagonists and therapy with fibrinolysin combined with anticoagulants and antibiotics are discussed.

1. A respiratory-deficient mutant strain of yeast was obtained from wild strain of Saccharomyces servisiae by treatment with 4-nitroquinoline N-oxide. Ultrastructure and function of the wild or mutant strains and the mitochondrial fractions isolated from these strains were examined by biochemical and electron microscopic analyses. 2. The frequency of the respiratory-deficient mutant strain in yeast induced with 10-6M 4-nitroquinoline N-oxide was about 40 %. 3. Respiratory-deficient mutant strain is incapable of reducing 2, 3, 5-triphenyltetrazolium chloride salt and to grow on lactate medium. In addition to this, the mutant has been found to have lost its ability to take up oxygen in sodium succinate and pyruvate. 4. 4.Nitroquinoline N-oxide in the concentration that induces a mutant of yeast cells or its kin inhibits the oxygen uptake in normal strain. 5. The normal strain of yeast is characterized by difference spectrum corresponding to cytochromes a+as, band c+Cll respectively, whereas, the mutant strain containes almost no cytochromes a+ as, band C1 but contains normal or increased amount of cytochrome c. 6. Mitochondrial fraction isolated from mutant strain has largely lost its ability to oxidize succinate. On the other hand, NADH-, lactate-and cytochrome c-oxidase activities are reduced by about 1/17, 1/7 and 1/8 of that of normal strain, respectively. 7. Succinate dehydrogenase activity of mutant strain is almost zero. Moreover, this activity is not affected on the addition of phenazine methosulfate. NADH dehydrogenase activity of mutant stran is about 1/2 of normal strain. 8. The variations in mitochondrial structure of normal and mutant strain in the stationary phase have been followed with the aid of electron microscopy. In contrast to the normal strain, the mutant strain revealed distinct morphological changes in mitochondria, especially, the lack of cristae in its interior. The results have been interpreted to indicate that the mutant induced by 4.nitroquinoline N.oxide has a character of cyto. plasmic mutant.

For the purpose of revealing whether or not hemoglobin synthesis is inhibited by the AMD, the author estimated the hemoglobin level of AMD treated anilmals by microspectrophotometer, and found that the hemoglobin levels of all the developmental stages of erythroid cells were not inhibited by the AMD. The data indicated that about one half of mRNA for hemoglobin is synthesized in the early stage of specialization with the supplementary synthesis at the later stages and all these mRNA is stable and insensitive to AMD.

For the purpose to confirm the existence of the stem cells of the myeloid and erythroid cells in the circulating blood and to have the information of its morphologic entity, the author conducted morphologic observations on the peripheral and bone marrow cells of the x-irradiated parabionts having non-irradiated partners joined by the vascular parabiosis devised by the author, and the following results were obtained. 1. The control rats exposed to 1000R x-ray did not show any sign of recovery of hemopoiesis in bone marrow even 8 days after irradiation. 2. In the bone marrow and spleen of the lethally irradiated animals having non-irradiated partner in parabionts, precursor cells of granulocyte and erythrocyte appeared first 4 days after conjugation irrespective of the days after irradiation, and the hemopoiesis was restored completely on the sixth to the seventh day. The results have indicated that the circulat. ing blood has the stem cells which can dedifferentiate and transform into the precursor cells of the myeloid and erythroid cells within 3 days under adequate conditions. 3. On the basis of the morphologic observations on the peripheral blood of the parabiosis and non-irradiated rats revealded non-specific cells, a discussion was made on the possibility that some atypical lymphoid cells can serve as the stem cells of the myeloid and erythroid cells.

The composition of total lipid extracted with chloroform-methanol from the AV12-induced tumor was investigated by thin-layer and paper chromatography. The content of lecitin and sphingomyelin was somewhat decreased and a cerebroside was characteristically detected in this tumor.

Correlation of molecular structure with biochemical functions of the plasma membrane of the microvilli of intestinal epithelial cells has been investigated by biochemical and electron microscopic procedures. Repeating particles, measuring approximately 60 Åin diameter, were found on the surface of the microvilli membrane which had been isolated or purified from rabbit intestinal epithelial cells and negatively stained with phosphotungstic acid. These particles were proved to be inherent components of the microvillus membrane, attached to the outer surface of its trilaminar structure, and were designated as the elementary particles of the microvilli of intestinal epithelial cells. Biochemical and electron microscopic identification of these elementary particles has been carried out by isolation of the elementary particles with papain from the isolated microvillus membrane, followed by purification of the particles by chromatographies on DEAE-cellulose and Sephadex columns. The partially purified particles containing invertase and leucine aminopeptidase are similar in size and structure to those of the elementary particles in the microvillus membrane. Evidence indicates that each of the elementary particles coincide with or include an enzyme molecule such as disaccharidase or peptidase, which carry out the terminal hydrolytic digestion of carbohydrates and proteins, respectively, on the surface of the microvillus membrane. Magnesium ionactivated adenosine triphosphatase and alkaline phosphatase cannot be solubilized with papain but remains in the smooth-surface membrane after the elementary particles have been removed. Cytochemical electron microscopic observation revealed that the active site of magnesium ion-activated adenosine triphosphatase is localized predominantly in the inner surface of the trilaminar structure of the microvillus membrane.

We fractionated the red cells of Japanese acatalasemic individuals (four individuals in two families) by SAss's method and counted the number of reticulocytes and determined the catalatic activity by manometric and perborate methods on each fraction containing reticulocytes in the descending order from the upper to the lower layers. We also incubated each of these fractions at 37°C for 4, 10,24 and 48 hours, to see the catalatic activity along with the maturation of reticulocytes. Heat stability of catalatic fraction separated by DEAE column was also examined. The results of the study may briefly be summarized as follows. 1. a. There was no parallel relationship between the number of reticulocytes and the catalatic activity in Japanese acatalasemic red cells. b. There could be seen no decrease in catalatic activity while the number of reticulocytes did decrease by the incubation. 2. Heat stability of crude catalatic fraction from acatalasemic blood is approximately the same as that of crude catalase fraction from normal blood.

An attempt was made to find out the nature of catalase coritained in the red cell, especially in the ghost, For this the red cell ghost isolated were washed several times with CO2-saturated water or deionized water and the catalase activity per gram protein of the ghost was estimated. It was found that despite several washings, the catalase activity/gram protein of the ghost do not decrease as compared with the activity of the original red cell solution, indicating the presence of catalase in the ghost. In the case of hypocatalasemic blood the catalase activity in the ghost shows similar behaviors as with normal blood cells. It is assumed theoretically that there are two kinds of catalase having different affinity to the red cell ghost. Namely, one that is readily released from the ghost and the other that has a strong affinity. The affinity of hypocatalasemic blood to the ghost seems to be somewhat weaker.