検索

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.

Eleven cases of malignant lymphomas were treated with a fibroblast-inhibiting agent, chloroquine, and of these, one case of lymphosarcoma, two of acute and chronic lymphocytic leukemia, respectively, and two of giant follicle lymphoma showed regression of the enlarged lymph nodes and also of the enlarged spleen in some of the splenomegalic patients. In contrast, the drug proved ineffective in two cases of reticulum cell sarcoma and Hodgkin's disease, respectively. The side effects of the drug were minimal, and three of the 11 cases complained of nausea, anorexia or palpebral ptosis, which disappeared by decreasing the drug dosage or combining ATP preparation. The tissue culture study of biopsied lymph nodes from lymphocytic leukemia showed inhibition of the growth zone in a medium containing chloroquine indicating a possibility of the drug action not only upon the stromal tissue but also upon the parenchymal tumor cell.

A fibroblast-inhibiting agent, chloroquine, used in the treatment of animal tumors led to a reasonably good result, and this approach was extended to the treatment of human cancers. Of histologically proven 54 cases, the drug was effective in 38, ineffective in 15, and unknown in one. It proved to be effective in all the patients who were treated for over 2 months with exception of terminal patients. Of the various malignant tumors treated, excellent therapeutic effects were obtained in patients with carcinoma of the lung and bladder. In the cases where the drug was effective there were a decrease of the size of tumors, fall of serum lactic dehydrogenase, increase of necrosis, inhibition of the stroma, as well as improvement of the symptoms and general condition. As to the mechanisms of the drug action, it would be necessary to consider of its anti-inflammatory and humoral effects upon the host in addition to its inhibitory action on the stromal connective tissue of cancers. The present chloroquine treatment appears to have its indication in inoperable cases, and pre- and post-operative cases, and for the prevention of reccurrence of tumors. Studies are currently in progress in our laboratory to discover more potent fibroblastinhibiting agents and on the combined chemotherapy of chloroquine and other anti-turnor agents. We are indebted to the Department of Urology of our University for the generosity to allow us to use the clinical data on patients with cancer of the urinary bladder.

In the course of experimental isovalthinuria induced by cholic acid, S35-methionine or S35-cystine administered was incorporated into urinary isovalthine in guinea pigs. Sulfur atom of cysteine seems to be utilized much better for isovalthine synthesis than that of methionine.

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.

Liver homogenates could synthetize S-(isopropylcarboxymethyl) glutathione (GSIV) from isovaleric acid and glutathione, and GSIV thus formed was cleaved into L-allo-isovalthine by kidney glutathionase preparation. Isovaleric acid-l-C14 incorporated into GSIV without prior cleavage by the in vitro system. The discrepancy of configuration between urinary and in vitro synthesized isovalthine was discussed.

We experienced a case of eosinophilic granuloma in soft tissue, and demonstrated its patterns of hydrolytic and oxidative enzymes histochemically. Neutrophils were rich in acid phosphatase and glucose-6-phosphate dehydrogenase. Eosinophils had much acid phosphatase and less other hydrolytic and oxidative enzymes. Lymphocytes showed weak reaction in all enzymes. Lymph follicles and histiocytes or fibrocytes had moderately oxidative enzymes. Small blood vessels and collagen fibers were rich in alkaline phosphatase and had a moderate amount of oxidative enzymes and acid phosphatase.

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.

FUKUTOME has once reported that isovaleric acid is an isovalthinuria inducer but isovaleric acid-1-C14 administered to a dog does not incorporate into urinary isovalthine and glutamic acid is most strongly labeled among acidic amino acids excreted. Recently, however, KUWAKI has found that liver homogenates of some animals can synthesize C14-labeled S-(isopropylcarboxymethyl) glutathione (GSIV) from isovaleric acid-1-C14 and glutathione, and that GSIV can be converted into isovalthine by kidney homogenate or glutathionase preparation4. For the elucidation of the above discrepancy, FUKUTOME's experiments were repeated by using isovaleric acid-methyl-C14 or-1-C14, and it was again found that these isotopic compounds did not significantly incorporate into urinary isovalthine.

An intestinal absorption test with the use of D-xylose has been performed on 19 patients including 3 of acute hepatitis, 7 of chronic hepatitis and 9 of liver cirrhosis, and the following results were obtained. 1) The 5 hr urinary excretion and 2 hr blood level of D-xylose tend to increase in patients of acute and chronic hepatitis with severer disorder of liver functions. 2) The standard deviations of the 5 hr urinary excretion and 2 hr blood level of D-xylose are larger in liver cirrhosis than in the other liver diseases. Those cases having severe disorder of liver functions are found to be diminished in 5 hr urinary excretion and 2 hr blood level of D-xylose. 3) A decrease in the absorption of D-xylose from the small intestine of liver cirrhosis might be caused by the diminished surface area of villi of the small intestine.

For the purpose to reveal the mechanism of the stimulated erythropoiesis in anemic condition, the author observed the numerical changes of the erythroblasts from normal rabbit bone marrow cultured under the environment of varied oxygen tensions, and revealed the following: 1. The erythroblasts incubated with air are increased after 24 to 48 hours and decreased gradually disappearing by 120 hours with a corresponding increase of erythrocytes. But no active proliferation of the stem cells or proerythroblasts is observed, all the cells have differentiated to erythrocytes. Hyperoxygen tension suppresses the increase of erythroblasts slightly, while hypoxygen tension stimulates the increase. Data suggest that the cell number destined to be ineffective erythropoiesis is regulated by oxygen tensions of the environment. 2. Basophilic erythroblasts are reduced in number from the beginning showing not any increasing tendency. The reducing rate is almost the same among those cultured under the hypo- and hyperoxygen tension, comparable to that incubated with air. 3. The hypoxygen tension brings about a marked increase in the number of orthochromatic erythroblasts with a decrease in polychromatic erythroblasts suggesting an accelerated cell differentiation, while the hyperoxygen tension elicits the suppression in the formation of orthochromatic erythroblasts with suppressed differentiation. Data also show the lack of denucleation mechanism in polychromatic stages in vitro differing from the case of the bone marrow of anemic animal.

Intestinal absorption tests with the use of D-xylose were conducted on 12 healthy Japanese subjects and the following results were obtained. 1) The mean value of the urinary xylose excretion within five hours after an oral administration of 25 g of D-xylose was 8.07 g and standard error of the mean was 0.11. The mean of urinary excretion was higher than most of previous reports. 2) The 5 hr urinary excretion after intravenous administration of 25 g D-xylose in normal subjects was almost equal to that reported by BUTTERWORTH et al. 3) The rate of D-xylose absorption from the intestine of normal Japanese subjects was higher than that in Europe, Canada and U. S. A. 4) The differences in the pattern of the intestinal absorption of D-xylose in normal individuals seemed to originate from different dietary habit continued over the period of many years, especially of carbohydrate contents.

An anatomical study was made to follow the degeneration of fibers by means of Marchi technique in cat after making experimentally lesion in Forel H field. As the results the following conclusions were reached. 1) The ipsilateral distribution of the degenerated granules was in the anterior sigmoid gyrus, caudate nucleus, putamen and globus pallidus, thalamic nuclei medial to the internal medullary lamina, substantia nigra, rubrocerebellar system, medial longitudinal fascicle system, mesencephalic and pontine reticular formation and medial lemniscus. 2) There was also contralateral distribution to the interpositus and dentatus nuclei of the cerebellum via brachium conjunctivum, to globus pallidus via supraoptic commissure, to subthalamic region and substantia nigra via supramammilary commissure, and to red nucleus via tegmental decussaion. 3) The degeneration is so extensive that the Forel H-field seems to be the cross road of the extrapyramidal system in association with brainstem activating system.

Some investigations have been done on the relationships between the swelling-shrinkage change, oxygen consumption and state of oxidation-reduction of pyridine nucleotides of mitochondria, and between the swelling-shrinkage change of mitochondrial structure by Ca2+ and accumlation of Ca45 in rat liver mitochondria. A parallel relationship is observed between the Ca2+ induced swelling and Ca2+ accumulation. Both of them require Pi but not Mg2+, ATP and exogenous respiratory substrates and are inhibited by respiratory inhihitors or uncouplers of oxidative phosphorylation but not by the inhibitors of phosphorylating respiration. In this case the Ca2+ is transported with the phosphate even in ice cold. Even in the presence of antimycin A, moreover, Pi-dependent Ca2+ accumulation and Ca2+ induced swelling can be overcome by addition of ATP, which are inhibited by oligomycin. In the presence of Pi, mitochondria show shrinkage by addition of Ca2+ before the high amplitude swelling, which is closely correlated to the electron ransport chain and phosphorylation process of mitochondria, and the pattern of the mitochondrial shrinkage is quite similar to that observed in the case of respiratory control by ADP in intact mitochondria. This shrinkage of mitochondria is inhibited by respiratory inhibitor or uncoupler of oxidative phosphorylation but not by the inhibitor of phosphorylating respiration. From these data, therefore, it is considerd that the Ca2+ accumulation and Ca2+ induced shrinkage-swelling of mitochondria require the energy of oxidative phosphorylation with respect to the initial step before the oligomycin block.