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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.

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.

Effects of sodium oleate and bovine serum albumin (BSA) on rat liver mitochondrial function and structure were studied by measuring oxygen uptake, 90° light-scattering, adenosine triphosphatase activity and pyridine nucleotides fluorescence. 1. The low concentration of oleate induced the uncoupling of oxidative phosphorylation and the scattering change of mitochondria. This action of oleate differed from that of oleate at a high concentration which induces the high amplitude swelling with respect to its physiological and biochemical properties. The degrees of reversal swelling (shrinkage) and of oxygen uptake induced by oleate in the presence of Pi and succinate were altered proportionately to the concentration of oleate, and the concentration of oleate to the shrinkage coincided with that of the maximal respiratory release. 2. Antimycin A or 2, 4- dinitrophenol prevented the oleate-induced mitochondrial shrinkage, but the treatment of these agents after prior incubation with Pi and succinate allowed the shrinkage, though the degree was small in its extent compared with that in the absence of inhibitors. On the other hand, oligomycin did not affect the shrinkage with oleate. 3. BSA protected the mitochondrial phosphorylation from the uncoupling action of oleate without showing any effect of its own. A complete reversal could readily be demonstrated by a sufficient amount of BSA from the uncoupling, structural changes, and oxidation of intramitochondrial pyridine nucleotides induced by oleate in a low concentration. 4. The oleate-stimulated latent ATPase activity was proportional to the oleate-induced shrinkage of mitochondria with respect to the concentration of oleate. The latent ATPase was abolished also by the addition of a sufficient amount of BSA. 5. The action of oleate on the phosphorylation sequence of mitochondria was discussed on the basis of the present findings.

For the purpose to reveal the mechanism of uptake and degradation of NAD by cells, the authors conducted the observation on the L cells cultured in the medium containing NAD and the following results have been obtained. 1. NAD in the medium is taken up by the cells in its intact form, reaching about twice the value of the control. 2. The spontaneously degraded products of NAD, nicotinamide and adenine dinucleotide ribose, in the same molar concentration as NAD used in the present experiment, have no effect on the NAD content of L cells. 3. The NAD taken up by the cells is degraded into nicotinamide mononucleotide (NMN) and adenine mononucleotide (AMP) by pyrophosphatase including NADpptase and excreted in the medium. Unexpectedly the ingested NAD is not degraded by NADase in the L cell. 4. L cells metabolize the same amount of NAD as that contained originally in the cell for about ten minutes, as calculated from the amount of NMN excreted in the medium.

An electon microscopic study on the structural differences among the red, white and intermediate muscle fibers of mice was made and the following results were obtained. 1. The red fiber contained very numerous mitochondria, the white fiber a few and the intermediate fiber a moderate number. The distribution of mitochondria was different in each type of muscle fiber. The cristae of mitochondria of the red fiber was quite well developed, that of the white fiber poorly and that of the intermedtate fiber moderately. 2. Sarcoplasmic reticulum of the white fiber was considerably well developed but that of the red and intermediate fibers poorly developed. 3. Glycogen particles were abundant in the white fiber, less in the intermediate fiber and least in the red fiber.

Respiration, activity of oleate oxidation and composition of the total fatty acids of rat liver were investigated in 3'-Me-DAB feeding. 1. Oxidative phosphorylation of rat liver mitochondria decreased temporarily at relatively earlier stages (about 2 to 3 weeks) in 3'-Me-DAB feeding. 2. The activity of oleate oxidation of rat liver mitochondria decreased rapidly to about one third of that in control groups after the start of 3'-Me-DAB feeding. 3. In the composition of the total fatty acids of rat liver, the proportion of oleic acid increased in 3'-Me-DAB groups. 4. Unknown octadecamonoenoic acid was observed in liver mitochondria of rat fed on 3'-Me-DAB. 5. Proportions of oleic and palmitoleic acids in liver tumors and mitochondria of liver tumors induced by 3'-Me-DAB feeding increased remarkably in contrast with decrease in those of palmitic and eicosapolyenoic acids. 6. A possibility was discussed about how higher level of oleate in the liver cells in azo dye feeding may be concerned with the tumor induction.

Sowohl aus dem Grunde, den Mechanismus des gestörten BTS-Stoffwechsels bei den Leberkrankheiten zu erkennen, als auch Beobachtungen für seine Behandlungen zu machen, wurden die Ein£liisse von Zucker, Thiamin und seinen Derivaten (BTMP, TTFD), Thioctsäure, der Verbindung zwischen dem Derivat von Thiamin und Thioctsaure (TATD), Kalium- und Magnesium-Asparaginat und Glucocorticoiden auf den BTS-Blutspiegel untersucht. Das führte zu folgenden Ergebnissen : 1) Der Anstieg des BTS-Blutspiegcls nach Belastung von Glukose bzw. Sorbit wurde beide Male beobachtet, aber er war nach Sorbit geringer als nach Glukose. Das bedeutet, dass Sorbit die BTS-Oxydation fördert. 2) Während der Anstieg des BTS-Blutspiegels mit Thiamin hydrochlorid nicht gehemmt wurde, wurde er mit Thioctsäure in vielen Fällen gehemmt, insbesondere in Rekonvaleszenz der akuten Hepatitis. 3) Nach der Verabreichung Von BTMP, TTFD und TATD war der BTS-Blutspiegel herabgesetzt, aber ihre Einwirkung war bei den Fällen mit gestörter Leberhämodynamik nicht gut. 4) Ebenso hat Kalium- und Magnesium-Asparaginat ungeFähr im Drittel der Fälle den BTS-Blutspiegel erniedrigt. Aber seine Einwirkung in Fällen mit gestörter Leberhämodynamik war ungünstig. 5) Der BTS-Blutspiegel wurde durch Glucocorticoide erhöht.

1) OX substance showed marked cytotoxicities in cell suspension culture of Yoshida sarcoma cells, celothelioma cells, and Ehrlich ascites tumor cells. It has become clear that the cytotoxicities have two aspects; one, nuclear shrinkage and karyolisis as seen with Carzinophilin and the other, cytoplasmic swelling as seen with Nitromin. 2) OX substance was effective by its contact action on patients with peritonitis carcinomatosa, celothelioma and rectal carcinoma. 3) Esterified OX substance was injected intravenously or intraperitonealy into CBA mice with ascites leukemia. The substance prolonged their life span and inhibited the progression of leukemia. As it was possible to give the substance repeatedly into mouse tail veins in this experiment, in the future, OX substance might become intravenously injectable for the treatment of patients with leukemia and solid malignant tumors.

1. For the purpose to clarify the relationship between the structural change and lipid composition of isolated rat liver mitochondria, lipid composition and swelling rate of mitochondria obtained from the rat of 3'-Me-DAB feeding and raised in cold room are measured, and the following results were obtained. 2. The mitochondria obtained from the liver of 3'-Me-DAB-fed rat and of rat raised in cold room show a low rate of swelling by addition of Na-oleate accompanied by the decrease in highly unsaturated fatty acids (C18:3 and C20:3or 4) and with the increase in saturated fatty acids (C16 and C18). 3. Activation energy for the mitochondrial swelling is about 16.2 Kcal in the mitochondria obtained from normal rat liver, but requires 19.7 Kcal in the mitochondria that show a low rate of swelling. The fatty acid composition, especially in glycerophosphatides which occupy about 80 per cent of total lipids, is a structural component of mitochondrial membrane, undergoes the change from former to latter in the following fashion: C16:0 21.73→32.10, C16:1 3.37→2.96, C18:0 25.0→29.75, C18:1 13.75→17.40, C18:2 23.90→16.0 and C20:3 or 4 12.23→1.79. 4. At the time of low rate swelling of mitochondria isolated from 3'-MeDAB- fed rat liver, there could be observed a marked increase of the acetone soluble lipid (simple lipids) in the total liver lipids and in the fatty acid distribution of the acetone-soluble lipids, oleic acid was markedly increased (0.838→3.81%/dry liver), despite the fact that in the acetone-insoluble fractions or in the mitochondria there are no marked changes in the oleic acid contents (1.84→2.56% or 0.212→0.246%/dry liver).

1. The forms of irritation causing inflammation and pain are reviewed, with reference to the significance of histamine, serotonin and bradykinin and in particular to the interrelationship between inflammation and pain. 2. The various types of experimental pain are reviewed and mention is made of the human and animal analgesia test methods derived from them. 3. More detailed descriptions are given of the analgesia test methods used by us, namely: a) Silver nitrate gonarthritis-pain, rat, in which both strong and weak analgesics with an anti-inflammatory action are effective. b) Phenylquinone-induced abdominal pain, mouse, in which all the analgesics and anti inflammatory agents mentioned in this article are effective in a greater or lesser degree. c) Tail-flick and hot-plate tests, mouse, in which the strong analgesics, the weaker analgesics and the anti-inflammatory agents, with the exception of the salicylates, are effective. d) Dental-pain test, guinea pig, which can be used to demonstrate the activity of the various analgesics, including the salicylates and also colchicine, which is not active in any other test. e) Pressure-pain, mouse, in which only the strong analgesics (narcotics) are effective. 4. The action of a large number of analgesics, anti-inflammatory agents and related drugs in the various analgesia-tests and in acute experimental inflammation is presented in tabular form. 5. It is concluded that the use of several pain and inflammation tests is essential for screening both analgesics for special indications (severe, mild pain, pain due to inflammation, etc.) and universal pain-killing drugs.

The swelling-shrinkage and oxidative phosphorylation of rat liver mitochondria affected by 3'-methy1-DAB feeding were observed in correlation with function by the method mentioned, and the following results were obtained: 1. By feeding 3'-methy1-DAB the swelling-shrinkage ability of rat liver mitochondria showed a remarkable alteration reducing in the amplitude. It reduced gradually during the days of feeding, reached the minimum value on 30th day and restored gradually thereafter (in Case 1). 2. ADP/O ratio also decreased by feeding the carcinogen reached the minimum point on 30th day and increased on 38th day showing the similar tendency in the decrease of the swelling-shrinkage amplitude (in Case 1). 3. The mitochondria from the hepatoma, which was induced by 3'-methy1DAB feeding, showed a lower amplitude in swelling-shrinkage with the dropped ADP/O ratio compared with those of mitochondria from liver tissue neighbouring the tumor. 4. The mechanism in the reduction of swelling-shrinkage ability has been discussed in the relation with fatty acid composition of mitochondria which is reported elsewhere. 5. From the above results it is deduced that lowered ability for swellingshrinkage with the reduced oxidative phosphorylation will be somehow related to the mechanism of cancer induction.