Search

The contents of nucleic acids in rat liver and hepatoma mitochondria and the physico-chemical properties on DNA's isolated from these mitochondria were comparatively investigated. The results are briefly summarized as follows. 1. The contents of DNA and RNA per mg protein of the hepatoma cell mitochondria were about 10 and 2 to 4 times higher than those of rat liver mitochondria, respectively. 2. The λ max. and λmin. values of DNA isolated from the hepatoma mitochondria were 257 mμ and 231 mμ, respectively and those of DNA isolated from the nuclei were 259 mμ and 233 mμ, respectively, in saline-citrate, pH 7.0. 3. Three fractions of mitochondrial DNA were obtained by the sucrose density gradient and these DNA fractions corresponded, probably, to about 30 S, and 20 S and 14 S DNA's. 4. There was little difference in base compositions between nuclear and mitochondrial DNA's of the hepatoma cells. 5. The degree of hybridization between the nuclear and mitochondrial DNA's of the hepatoma cells was almost the same as that between the nuclear and nuclear DNA's of the hepatoma cells, and somewhat higher than that between the nuclear DNA of rat liver and the nuclear DNA of hepatoma cells. 6. "Highly twisted" circular, "open" circular and linear forms were observed in the DNA preparations of the hepatoma mitochondria. The average values of contour lengths of rat liver and the hepatoma DNA's observed at high frequency were 5.3 μ and 4.5 μ. 7. A discussion was made on the relation between the genetic informations of mitochondrial DNA and the formation of a mitochondrion in rat liver and the hepatoma cells.

As a link in a series of studies on the effects of blood constituents on the brain function by means of brain perfusion, we used four kinds of artificial blood; namely, the blood containing a low molecular dextran, one containing glutamic acid, one containing essential amino acid group and the one containing both essential amino acid group and glutamic acid. During the perfusion experiments we observed the effects of blood constituents on the function and metabolism of the perfused brain and obtained the following results. 1. When a low molecular dextran is used as the colloid osmotic pressure agent instead of hydrodextran, the amount of the blood flow in the brain is maintained roughly at a certain fixed level throughout the experiment, showing no gradual decreasing tendency. 2. When using the artificial blood supplemented with glutamic acid, EEG of the perfused brain shows an increase in the appearance rate of β32 and β33 bands, approaching closely to the pattern of EEG of unrestrained controls at arousal state. 3. In the case of the blood added with essential amino acids similar to the case using the blood with glutamic acid, EEG approaches towards the alert pattern of the controls. 4. When the perfusion is done with the artificial blood lacking in amino acids, about one hour after the start of the perfusion the amount of glutamic acid and its related compounds in the brain can no longer be maintained at normal level and the decrease, being so marked, brings about a marked decrease also in total amino acid content. 5. When the perfusion blood contains glutamic acid, essential amino acid group or both, the concentrations of amino acids of the brain glutamic acid group and the total amino acid can be maintained approximately at normal level for the duration of over one hour.

An electron microscopic study on the fine structural differences of motor endplates among the red, white and intermediate muscle fibers of the rat intercostal muscles was made and the following results were obtained. 1. In the motor endplate of the red fiber, the junctional folds were poorly developed and their number was small. 2. In the motor endplate of the white fiber, the junctional folds were well developed and their number was far more numerous than those in the red fiber. 3. The fine structure of the motor endplate of the intermediate fiber was of an intermediate character between the red and white fiber.

Since Hahn's observation of the postalimentary lipemia clearing actIvity following the injection of heparin, physiological, biochemical and clinical significances of the postheparin lipoprotein lipase have been well clarified. The presence of the endogenous lipoprotein lipase in human blood, which was at first doubted, has been repeatedly confirmed2∼8. Recent papers9,10 described elevated endogenous lipoprotein lipase activity in patients with essential hyperlipemia after ample fat uptake. In this preliminary report, changes of the lipoprotein lipase activity during oral glucose tolerance test is illustrated.

1. An oligomycin -sensitive ATPase was isolated and partially purified from beef heart mitochondria. The specific activity of ATPase sensitive to oligomycin of the fraction was five to eight times that of aged mitochondrial or of DNP-induced mitochondrial ATPase assayed under the same condition. 2. Electron micrographs of the partially purified oligomycin- sensitive ATPase reveal a structure in which headpieces are regularly attached by way of stalks to a thread-like structure derived from a superficial portion of base pieces. 3. A high concentration of the structured material coincided with a high activity of oligomycin-sensitive ATPase. When the headpieces were detached from the structure, the ATPase became insensitive to oligomycin. 4. The fraction of oligomycin -sensitive ATPase was essentially free of membrane structure and was contaminated with a small amount of cytochromes b and Cl but no cyt. a. Cytochrome concentrations of the preparations were indifferent to the activity of oligomycin sensitive ATPase. It follows that ATPase does not require cytochromes or membrane structure for its oligomycin sensitivity. 5. From these results it seems that the factor rendering ATPase sensitive to oligomycin should be contained in the stalks and/or the thread-like portion of basepieces of the structure. The structure is the simplest unit of oligomycinsensitive ATPase as yet obtained. 6. The structure was called "oligomycin-sensitive ATPase particles" (abbreviated as OSA particles). A unit of OSA particles consists of a headpiece attached by a stalk to a portion of base piece.

Non-hemin iron content in gastric juice was examined in 46 patients with various blood diseases, especially idiopathic hypochromic anemia and in 26 healthy controls. 1. The iron content in gastric juice was found to be 290 μg/ dl in healthy controls, a lower value of 110 μg/ dl in idiopathic hypochromic anemia and a higher value of 550 μg / dl in aplastic anemia. These values were in a close correlation with serum iron or sideroblasts. 2. In idiopathic hypochromic anemia there was also a close correlation between the iron content in gastric juice and hemoglobin. In the course of treating idiopathic hypochromic anemia (stage of recovery of anemia) the iron content in gastric juice showed a marked increase over the value in healthy controls as well as a transient increase after an intravenous iron tolerance test. This condition may be interpreted as an "iron-losing anemia". Iron excretion of gastric mucosa in various blood diseases and its changes in the course of treating idiopathic hypochromic anemia in relation to the cause of this anemia were discussed.

The site of localization of TCA cycle dehydrogenases in mitochondria has been investigated by observing the dehydrogenase activities and fine structure of the fractionated samples after freezing and thawing or sonication of beef heart and rat liver mitochondria. 1. In the sonicated mitochondria, activities of malic and isocitric dehydrogenases were highest in the supernatant fraction centrifuged at 198,000 x g for 60 minutes, while the specific activity of a-ketoglutaric dehydrogenase was higher in the fluffy or residue fraction. The distribution of the activity of pyruvic dehydrogenase was similar to that of a-ketoglutaric dehydrogenase. 2. In a sucrose density gradient fractionation of the fluffy fraction obtained by centifugation of sonicated mitochondria at 198, 000 x g for 60 minutes, the activities of malic and pyruvic dehydrogenase were observed in the top (or low density) layer in the form of fine particles, while that of a-ketoglutaric dehydrogenase was observed in the middle (or medium density) layers in the form of aggregates of fine particles and membranous fragments. 3. In the samples fractionated after freezing and thawing of mitochondria, which were considered to be a relatively mild disruption, the specific activity of a-ketoglutaric dehydrogenase was higher in the residue (submitochondria) fraction than that in the supernatant fraction (centrifuged at 144,000 x g, 30 minutes), and the activity of malic dehydrogenase still remained significantly high in the residue fraction. 4. It was deduced that the TCA cycle dehydrogenases could be localized in the matrix of the mitochondria by a loose binding to the inner membrane.

The results of our study may briefly be summarized as follows: 1) The irradiation with microrays (20∼30 watts) similar as 2,000 R and 5,000 R Gamma radiation did not substantially affect the activity of fibrinolysin (SK+SD). 2) By the irradiation method so far mentioned it has been demonstrated that the fibrinolytic activity of anticoagulant of the SK+SD preparation is preserved in all the clotting systems which we used. 3) Our findings indicate that it is possible to irradiate patients for therapeutical purpose with Radarmed (electromagneticrays) provided that there is produced some enhancing influence of the same blood clotting factors or systems. Together with earlier works in this field it appears that this method of the microirradiation could provide us with an important evidence on which we can base our further in vitro and in vivo radiohematologic studies; investigations with various preparations, types of radiation that are still underway9∼16.

With a certain fixed methods of analyses, we carried out the determination of flavins and cytochromes in the mitochondria (Mt) and electron transfer particles (ETP) of the heart and liver of rats and cows, and made a comparison of the data with one another. Our findings may briefly be summarized as follows. 1. The concentration of each component of the beef heart mitochondria proved to be 0.47 for acid extractable flavins; 0.22 for acid nonextractable flavin; O. 75 for cytochrome (cyt.) a; 0.58 for cyt. b; and O. 51 for cyt. C + Cl, all units being mμ mole per mg of protein. 2. In the beef liver mitochondria it was 0.46 for acid extractable flavins; 0.18 for acid non-extractable flavin; 0.092 for cyt. a; 0.089 for cyt. b; and 0.122 for cyt. C+Cll likewise all units in term of mμ mole per mg of protein. 3. In the case of rat heart mitochondria, it was found to be O. 42 for acid extractable flavins; 0.22 for acid non-extractable flavin; 0.88 for cyt. a; 0.41 for cyt. b; and 0.62 for cyt. C + Cll all in mμ mole per mg of protein. 4. In the rat liver mitochondria it was 0.56 for acid extractable flavins; 0.19 for acid non-extractable flavin; 0.20 for cyt. a; 0.14 for cyt. b; and 0.19 for cyt. C+Cl. 5. The concentration ratios of Fs, cyt. a and cyt. b of the mitochondria, what are considered to be intrinsic and fixed components of the mitochondrion. to those of the electron transfer particles were 1. 3 in both the beef heart and the rat heart, while 2.2 in the beef liver and 2.1 in the rat liver. 6. These findings were compared with the data reported by other workers, and also a discussion was made on the molecular organization of the mitochondrial inner membrane.

For the purpose to know whether the annual increase of leukemia incidence in Japan is due to some leukemogenic factors or due to the increased detection rate, the authors made some statistical survey of autopsy cases in which the diagnosis is reliable and not any type of leukemias escape the detection. The results showed that acute leukemias, which are found mostly in younger age, is actually increasing. In addition, it has been deduced that among the suspected factors the increase in ionizing radiation will be one of the most probable factors for the increase in leukemia incidence

In order to know the precise quantity of catalase protein in acatalasemic and hypocatalasemic blood, immunological studies were conducted using hemolysates or acetone extracts of those blood as antigen. 1) The ratio of catalase contained in normal, hypocatalasemic and acatalasemic blood, calculated from precipitates produced in the reaction between catalase antibody and hemolysates was 1.0 : 0.5 : 0.07. 2) The ratio of catalase in normal, hypocatalasemic and acatalasemic blood, calculated from precipitates from the catalase antibody and the acetone extracts was 1.0: 0.49 : 0.11. In the precipitin ring tests using acetone extract, the antigen titer in normal, hypocatalasemic and acatalasemic extracts was 40, 20, and 0 respectively. 3) From our experiments it can be said that hypocatalasemic blood shows one half the catalase activity of normal blood, due to one half the quantity of catalase protein, and that acatalasemic blood lacks catalase activity due to the absence of the catalase protein. These findings strongly suggest that no substances exist which suppress or inhibit the catalase activity in hypocatalasemic and acatalasemic blood.

It was investigated to clarify the relationship between the composition of the lipid fractions obtained from the ascitic fluid of Ehrlich ascites tumor bearing mice and its uncoupling activity after whole body irradiation (1,000 r). 1. Oxidative phosphorylation of Ehrllch ascites tumor cells was loosely uncoupled with the addition of ascitic lipid fraction extracted from tumor bearing mice. 2. The uncoupling activity of the lipid fraction on the oxidative phosphorylation of the tumor cells increased after whole body irradiation. 3. Ascitic lipid fraction, especially acetone soluble fraction accelerated mitochondrial swelling, and the swelling action was increased remarkably by the whole body irradiation. 4. No significant changes were observed in the proportion of acetone soluble fraction to acetone insoluble fraction in the ascitic lipid after X-irradiation, and the proportion of the both fractions was approximately 9 : 1, respectively. 5, Main compositions of total and non-esterified fatty acids in the ascitic fluid obtained from the control and X-irradiated groups were palmitic, stearic, oleic, linoleic and palmitoleic acids, and the proportions of unsaturated acids, especially oleic and linoleic acids in both fatty acid fractions were greater in the X-irradiated group. 6. Remarkable increment of unsaturated fatty acid especially linoleic acid, was also observed in the total fatty acids of the tumor cells separated from the X-irradiated group. 7. It can be concluded that an uncoupling agent extracted from ascitic fluid of the X-irradiated group was a mixture of long-chain fatty acids, especially oleic and linoleic acids. 8. It was also discussed that uncoupling oxidative phosphorylation in liver mitochondria after whole body irradiation may be caused by a similar mechanism to that in the turnor cells.

Bei 365 Leberpatienten wurde die BTS im Blut bestimmt und bei einem Teil der Fälle wurde die Leberkatheterisation durchgefiihrt. Das fiihrte zu folgenden Ergebnissen : 1) Der BTS-Blutspiegel war ungefähr in 50 Prozent der Fälle erh&#öht, und zwar fallend in der Reihenfolge bei Leberzirrhose, akuter Hepatitis und chronischer Hepatitis. Bei der akuten Hepatitis war er in der Rekonvaleszenz erhöhter als im akuten Stadium und auch bei chronischer Hepatitis war er bei längerem Krankheitsverlauf höher als bei krzem. 2) Eine Parallelitat zwischen dem BTS-Blutspiegel und der Müdigkeit bestand, besonders war er bei den Fällen, die durch routine Leberfunktionsproben normal beurteilt wurden und in denen über Müdigkeit geklagt wurde, erhöht. 3) Eine Korrelation zwischen dem BTS-Blutspiegel und den routine Leberfunktionsproben bestand nicht, doch bei den Fällen mit normalen routine Leberfunktionsproben fiel der BTS-Blutspiegel in 46.5 Prozent der Fälle positiv aus. Es wäre denkbar, dass der BTS-Blutspiegel eine von den routine Leberfunktionsproben nicht ergriffene Seite ausdrticken könnte. 4) Es gab keine Korrelation zwischen dem BTS,Blutspiegel und dem histologischen Befund der Leber. Doch durch Laparoskopie war der erhöhte BTS-Blutspiegel insbesondere bei dem Ⅱ. und IV. Typus der grossen weissen Leber festgestellt worden. 5) Weil L·BTS in Fällen erhöhter V-BTS (d. h. BTS im Blut) höher war als V-BTS bzw. A-BTS, ausserdem die engste Beziehung von L-BTS/ABTS- Quotienten zum V-BTS-Spiegel (d. h. der BTS-Blutspigel) bestand, wurde bestätigt, dass die erhöhte BTS im Blut bei Leberkrankheiten aus Leber stammt. Der BTS-Blutspiegel spiegelt nämlich den BTS-Stoffwechsel in der Leber wider. 6) Der BTS-Stoffwechsel in der Leber stand in engster Beziehung zu der Leberhamodynamik, d. h. zum visceralem Sauerstoffverbrauch, zum geschatzten Leberdurchblutung und zum Lebervenenverschlussdruck.

1. In the absorption spectra of crude catalase solution (Stages 2, 3, and 5) of normal blood, three absorption bands characterizing catalase molecules are recognized. 2. The three absorption bands specific for catalase cannot be found in acatalasemic blood extracts (Stages 2 and 3). 3. It is inferred that catalase is not present in the crude catalase extract from acatalasemic red blood cells.

An experimental study was attempted to make an analysis of the subcortical and brain stem lesion effect on the Metrazol-induced corticogenic epileptic convulsion based on EEG-discharge and EMG-convulsion as indicators. utilizing 42 adult cats. 1. A definite threshold increment of eliciting the seizure was found in the case of bilateral lesion of the Forel H-field. In contrast to it, no variation in the threshold was found in the case of the lesions at the other parts of brain stem, thalamus, red nucleus and its neighborhood, and lenticular nucleus. 2. There was a parallel relation between EEG discharge and convulsion. Dissociation could be obtained in none of the cases. 3. It is, therefore, to be concluded that the Forel H-field is composed of the main axis of cortico-subcortical reverberating circuit and that the lesion causes a decrement of the excitability at cortex and an inhibition of the corticogenic epileptic convulsion.

1. An apparatus for the simultaneous measurements of volume change, fluorescence intensity of pyridine nucleotides and oxygen consumption of mitochondria has been constructed. 2. Oxygen consumption is measured by the rotating platinum electrode with a modification of Hagihara's system, attached in a cuvette of the apparatus. 3. Volume changes of mitochondria (swelling-shrinkage) are measured by the 90° light-scattering at 650 mμ. 4. Relative fluorescence intensity of pyridine nucleotides is measured by the fluorometer: for the excitation, a bright light at 365 mμ. line of mercury lamp is isolated through the filter and exposed to the mitochondria suspended in a cuvette of the apparatus, and fluorescent emission is analyzed by a grating mirror monochromator. 5. The scattered light at 650 mμ. is not affected by the excitation light and the fluorescent emission, and fluorescence intensity is not affected by the scattered light at 650 mμ. 6. The simultaneous measurements of the oxidation-reduction of pyridine nucleotides, the respiration states and the changes in the intensity of 90° lightscattering of mitochondria are given as an example of the performance of the present apparatus.

An electron microscope study was performed on the ultrastructure and developmental process of the Mukai strain of Japanese B encephalitis virus propagated in vitro on porcine kidney stable cells. The virus particle of Japanese B encephalitis is hexagonal in sections and approximately 40 mμ in the maximum diameter, composed of an outer membrane, 20Å thick, viroplasm, 30 Å thick and an electron-dense nucleoid, 25 mμ in diameter. The virus particles develop by a budding process on the wall of the cytoplasmic vacuole. Thereafter, virus particles are densely packed in the vacuole usually in random arrangement and rarely in crystalline arrays. The vacuole containing virus particles gradually moves toward the cell surface and liberates the virus particles to the exterior of the cells through a narrow canaliculus. A structure suggestive of incomplete virus particles was also observed.

For the investigation of iron metabolism in the intestinal mucosa in various　blood diseases, intestinal biopsy (duodenum) was performed on 10 healthy controls　and 35 cases with various blood diseases. The following are　the results　of the studies on distribution of stainable　iron, amounts of non-hemin iron in the　biopsied materials, and iron uptake of the intestinal epithelial cells.　1) An evaluation of distribution of stainable iron by Berlin　blue reaction　showed none or very mild degree, if any,　inhealthy controls, an increase in　aplastic anemia,　pernicious anemia, some of leukemias and in iron deficiency anemia following iron therapy, and a decrease in　idiopathic hypochromic anemia, anchylostomiasis anemia,　anemia with cancer, myxedema, hemolytic anemia,　and in　some of leukemias. Some of anemia with cancer, however,　showed an　increase of a certain degree. In iron absorption tests, no changes were found other than a very mild increase in aplastic anemia. 2) Non-hemin iron was 70-112γ/g in healthy controls, increased in aplastic anemia approximately to 100-200γ/g, ranging 40-130γ/g in leukemia, and decreased in idiopathic hypochromic anemia and in anemia with cancer ranging 30-60γ/g and 30-50γ/g respectively. Amounts of non-hemin iron and serum iron or sideroblasts show a fair correlation. The fractionation of nonhemin iron in aplastic anemia didn't show any difference in relationship of each fraction from healthy controls despite the increased amount in the former. 3) A radioautographic evaluation of iron uptake by intestinal epithelium was performed by our device for evaluation of intestinal absorption capacity. The iron uptake was mild in healthy controls, almost none in aplastic anemia, and marked in iron deficiency anemia where it was decreased approximately to the level of healthy controls following iron therapy. 4) The intestinal tissue iron showed a series of changes similar to those of iron present in the serum or erythroblasts, and the non-hemin iron in the intestinal mucosa is inversely correlated with iron uptake of epithelium and is considered to regulate the absorption according to its amount.

For the purpose to look into the regulatory mechanism of erythropoiesis, changes in the cell volume and the cell size of the erythroid cells have been observed in peripheral blood and marrow from normal and phenylhydrazine induced anemic rabbits. And the following results have been obtained: 1. After the injection of phenylhydrazine hydrochloride a hemolytic anemia can be induced with a marked increase in the reticulocyte number. The cell volume increases with the advance of anemia but it is never proportional to the increase of reticulocyte number. The MCV reaches the value twice the normal but it never exceeds the threshold. 2. In bone marrow the smaller sized orthochromatic cells are reduced extremely in number or obliterated in anemic animals. As there is not any marked difference in cell size of polychromatic erythroblasts between normal and anemic animals, the large red cells of anemic animal will be formed by denuc1eation of the polychromatic erythroblasts. 3. The percentage of basophilic erythroblasts is increased in anemic animal suggesting an accelerated differentiation-division of proerythroblasts to basophilic ones. 4. The data strongly support the denuc1eation at polychromatic stage in emergency but not at the younger stages than polychromatic erythroblast. Data also suggest that in severe anemia an accelerated cell division occurs, especially in the stage from proerythroblast to basophilic erythroblast.

Ehrlich ascites tumor cells affected by oleic and linoleic acids lose their cytomembrane followed by the leak out of ribosomes. Some cells survived through this treatment when they were transplanted into mouse peritoneal cavity, but they changed their characteristics showing wider and less basophilic cytoplasm and smaller nuclei with dense nuclear chromatin and ambiguous nucleoli. In spite of many attempts, no qualitative changes have been found between normal and cancer cells. Recently, Ishikawa found the specific antigenicity of cancer cell membrane which was common to several strains of canccr cells. Grobstein and coworkers have clarified that pancreatic cells can differentiate in association with neighboring mesenchymal cells, probably getting some information. Their works suggest that the cell differentiation will be induced by mutual association of cells by which the cell will receive some substance acting as the information for differentiation. Taking the works of Ishikawa and his collabolators into consideration, it seems that cancer cells may be unable to differentiate by their defective or incomplete cell membrane through which they cannot associate with neighboring cells and fail to get the information. Almost all of the biological characteristics of cancer cells, immaturity, autonomic growth, invasive and metastatic properties independent from the neighboring cell groups, are well explained or consistent with this view. Recently, we found that the cell membrane can be loosened by some unsaturated fatty acids resulting in the leak-out of ribosomes. In this paper it is demonstrated how the Ehrlich ascites tumor cell affected by fatty acids lose their cytomembrane and the ribosomes and how the cells survived through this treatment show different characteristics from the original ones, taking the appearance more matured cells.