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1. Both EACA and AMCHA clearly showed an anti-inflammatory effect, by intravenous, intramuscular, or oral route, against inflammatory edema produced in rats by intracutaneous injection of rabbit's anti-rat serum, carrageenin, histamine, serotonin, or bradykinin, as tested by the punch method. 2. The two compounds also showed inhibitory action against the cotton pellet granuloma when used in a larger dose. 3. There was virtually no difference between the two compounds in their anti-inflammatory activity, in spite of the fact that antiplasmin activity of AMCHA is evidently greater than that of EACA. In addition, there was no increase in fibrinolysis at the site of antiserum inflammation in rats. Therefore, it would be difficult to presume that the anti-inflammatory action of these compounds is due to their antiplasmin activity. 4. Salicylates, pyrazolidine derivatives, and non-steroidal antiinflammatory agents like flufenamic acid inhibited degranulation of isolated rat mast cells induced by compound 48/80 and also inhibited ATP-32Pi exchange reaction in rat liver mitochondria, but such actions were not observed in EACA or AMCHA. 5. Anti-inflammatory effect of EACA and AMCHA did not decrease after adrenalectomy but did become weak in hypophysectomized rats. EACA did not increase blood sugar level in normal rats so that its antiinflammatory action is not due to hyperglycemia, and the effect of hypophysectomy may not be correlated to carbohydrate metabolism. 6. Anti- inflammatory effect of EACA and AMCHA appeared more rapidly after intramuscular or oral administration than by intravenous injection but the effect was not fortified after their in vitro incubation with tissues of stomach, intestine, or liver.

For the purpo3e to determine exactly what stage of cell specialization the DNA level of erythroid cell nuclei begins to decline, the author observed the DNA level of erythroblasts in mitosis by microspectrophotometry and the DNA synthesis by flash labeling with H3-thymidine. The cell samples were obtained from the bone marrow of normal, blood-depleted and phenylhydrazine-treated animals, and the anemic animals received a mass red cell transfusion, all the animals being injected with colchicine 4 hours before obtaining the bone marrow sample. DNA level was measured on the smeared cells stained by Feulgen reaction and DNA synthesis by autoradiography on the smeared cells. Besides these, chromosome number was observed on the anemic rat erythroblasts at metaphase by air dry method. The observations indicated that the DNA level begins to decrease at polychromatic stage being accompanied by a decrease in TDH3-incorporation into DNA, reaching minimum level at orthochromatic cell both in DNA contents and synthesis. Chromosome numbers of erythroblasts of rat were irregular being distri buted between 42 to 20. The data have suggested that the DNA level of erythroblasts decreases only in the later stages of cell specialization, and at polychromatic stage the chromosome number may also decrease in rabbit at polychromatic stage by the cell division with an incomplete DNA replication. The high DNA level of the erythroblasts of rabbit, in severe anemia where most of the cells are denucleated at polychromatic and late basophilic stages, has been discussed from the view point of the insufficient DNA replication at polychromatic stages.

1. The ratios of free 5α-cholestan-3β-ol and cholesterol and esterified 5α-cholestan-3β-ol were higher in pylorus than in cardia. 2. Esterified cholesterol level was higher in cardia than in pylorus. 3. Among the stomach cancer tissues examined free cholesterol level was higher than in the non-cancerous. 4. Esterified 5α-cholestan-3β-o1 and cholesterol levels were lower in the cancerous tissues than in the non cancerous.

1. Isovaleric acid-1-C14, -4-C14, or C14-CaC03 with or without non-isotopic isovaleric acid was orally administered to rats and the incorporation of these isotopes into liver cholesterol, fatty acid, or urinary isovalthine was examined. 2. Isopropyl group of isovaleric acid was more efficiently utilized for cholesterol synthesis than carboxyl group, and also for cholesterol synthesis than for fatty acid. These results indicate that isovaleric acid is cleaved into two fragments before it is utilized for cholesterol synthesis. 3. Carbon dioxide was used for the synthesis of liver cholesterol and of liver fatty acid. Isovaleric acid seems to enhance the incorporation of carbon dioxide into cholesterol. 4. All the experimental rats received isotopic or non-isotopic isovaleric acid excreted isovalthine, but no radioactivity was found in it. Thus, isovaleric acid residue of urinary isovalthine molecule is not derived from isovaleric acid administered, and carbon dioxide is not the carbon source of urinary isovalthine. 5. Suspicious metabolism of isovaleric acid or of carbon dioxide was discussed. 6. Isotopic isovalthine which was synthesized from (± ) α-bromoisovaleric acid-4-C14 is administered to rat and it was found that the isotope did not incorporate into cholesterol or fatty acid of liver and of brain. About 15% of isotopic isovalthine was recovered in urine up to the next day after injection. The large part of isovalthine was missing.

For the purpose to clarify whether minimal catalatic activity exists in Japanese acatalasemic cells or not and the manner how extrinsic hydrogen peroxide affects the acatalasemic cells, the author performed tissue cultures using the skin specimens from four acatalasemic persons affected with Takahara's disease and studied the nature of these cultured cells. The results are summarized as follows. 1. Between normal and acatalasemic cultured cells, no morphological differences could be seen and the growth rate of these cell-lines was similar to one another. 2. On the activity of succinoxidase and cytochrome oxidase there could be observed no difference between normal and acatalasemic cells. 3. In each acatalasemic cell line the minimal catalatic activity was observed and it seemed that this activity has an important role in decomposing hydrogen peroxide under normal metabolic pathway. 4. After treating with 10-4M hydrogen peroxide, respiratory enzyme activities and the growth rate in the acatalasemic cells were markedly disturbed, while in normal cells these remained almost intact. 5. There could be observed no differences between normal and acatalasemic cultured cells after X-ray irradiation (200 to 600 r) on the succinoxidase activity, catalatic activity and growth rate.

The appearance of sideroblasts in hypoplastic anemia (HAl and acute myelocytic leukemia (AML), together with their sideroblastograms, was studied. Hematological studies on cases with type III sideroblast dominance by sideroblastograms produced the following results. Type III sideroblast dominant HA was observed in three of 63 cases. Two of the above three cases had what we call "atypical factor", while the remaining one became AML in its clinical course and could be considered to be leukemia in a hypoplastic preleukemic stage. Type III sideroblast dominant AML was noted in five of 32 cases. Three of these five cases are compatible with low percentage leukemia, and one of the above three cases showed ringed sideroblasts exhibiting erythroleukemia in the terminal stage. In HA and AML, type III sideroblast dominant cases have to be examined in relation to atypical HA and atypical leukemia. Changes of iron meta. bolism in erythroblasts with preleukemic stage will be attributable to disturbance of erythropoiesis such as erythroid hyperplasia in bone marrow and also, in part, to disturbance of hemoglobin synthesis.

The control mechanism of mitosis in the regenerating rat liver was studied in relation to the cell functions. Partial hepatec· tomy induces a series of changes prior to the initiation of mitosis, i. e. decrease in serum glucose and albumin levels, loss of glycogen from liver cells, and increased lipid mobilization to liver cells. Massive supplies of glucose and fructose suppressed significantly hepatocellu. lar mitosis with suppression of lipid accumulation and preservation of glycogen in the liver cells and of blood sugar level. Homologous serum administration also suppressed the rate of liver cell mitosis after hepatectomy preventing the decrease in serum albumin level, but did not suppress the lipid accumulation in the liver. Starvation, which would relieve the liver cell from the work of detoxication of intesti. nal toxic products, did not show any suppressive effect on the mitotic rate of liver cells after partial hepatectomy in single animals. But starvation induced severe hypoglycemia, moderate hypoalbuminemia and loss of glycogen content in the liver. These changes in metabo. lism by starvation and partial hepatectomy were suppressed by con· jugating the animals with nonhepatectomized fed.partners by aortic anastomosis, and mitosis was suppressed in the residual liver of the fasting animals in this parabiosis. The results indicate that all the major functions of parenchymal live cells tested, sugar metabolism, serum albumin production, and detoxication, are closely related to the control of liver cell mitosis. Accumulation of lipids in the liver remnant after partial hepatectomy is thought to be for the compensa. tion of reduced glycogen storage and not concerned directly with the liver cell mitosis. Discussion was made briefly on the humoral factor and portal blood factor in relation to excess load of functions on resi. dual liver cells.

Folic acid contents of plasma, whole blood, liver and spleen in Rauscher leukemic mice were estimated. Plasma and liver folate in the leukemic mice was lower than that of normal mice, suggesting that folic acid was deficient in Rauscher leukemia. Folic acid contents in whole blood and spleen were even higher in the leukemic mice than those in normal mice. Clearance study by injecting folic acid intravenously into leukemic mice showed faster disappearance of folic acid from the circulating blood, suggesting that folic acid demand of Rauscher leukemia is increased. Methotrexate administered shortly after inoculation of the virus did not prevent Rauscher leukemia. But anemia, reticulocytosis and erythroblastosis, which are commonly seen 3-4 weeks later in leukemic controls, were not marked as compared with controls. It can be concluded that the requirement of folic acid is greater in Rauscher leukemia than in controls, and methotrexate is effective for preventing hematological changes commonly seen in this type of leukemias.

The effects of cepharanthin (Ce), glycyrrhizin (G), verapamil (V), and G plus V on induced thermotolerance in NIH3T3 cells were studied. Cells were heated with or without the drug at 45 degrees C for 20 min (the first heating), incubated at 37 degrees C for 12h (the incubation period), and heated again at 45 degrees C for 0-210 min (the second heating). G and V were added throughout the experiment, while Ce was added throughout the experiment or during only the first or second heating, or the incubation period. The cells were harvested after the second heating to evaluate cell survival. In control experiments without any drug, thermotolerance developed and reached the highest peak in the cells incubated for 12h at 37 degrees C. However, thermotolerance in the control cells was suppressed by incubating them at 0 degree C, but developed by subsequent incubation at 37 degrees C. This suggests that the acquisition of thermotolerance by the cells required metabolic processes during the incubation at 37 degrees C. When each drug was present throughout the experiment, only Ce or the combined use of G and V was effective in reducing thermotolerance. Thermotolerance was also suppressed in the presence of Ce during the second heating. These results indicate that Ce reduces thermotolerance by enhancing thermosensitivity rather than by inhibiting the development of thermotolerance.

The influences of ventricular pacing at a rate of 70 beats/min (bpm) on the systemic and coronary hemodynamics, myocardial metabolism, and cardiac work efficiency were evaluated in five patients with bradycardia. The results were compared to those obtained in six normal subjects at rest. In order to elucidate the effects of a relatively high rate of ventricular pacing, cardiovascular and metabolic variables were also obtained at 120 bpm in the normal subjects. It was observed that the patients eventually benefited from ventricular pacing at a rate of 70 bpm and improved in systemic hemodynamics. Although coronary hemodynamics and myocardial metabolism were accelerated, the cardiac work efficiency was not improved. A pacing rate of 120 bpm in the normal subjects did not appear to accelerate systemic hemodynamics, but adverse accelerations of coronary hemodynamics and myocardial metabolism were observed, and the cardiac work efficiency was remarkably reduced as a result. Our observations indicated that the coronary reserve capacity was very important for ventricular pacing, and suggested that an undue increment of the pacing rate not only might be meaningless but also might induce ischemic angina. Therefore, we should be cautious in using a rate-responsive pacing mode, particularly in determination of the upper limit of pacing rates, although many benefits with this pacing mode have recently been advocated.

To test the hypothesis that the endothelium-derived relaxing factor (EDRF) contributes to coronary vasodilation induced by myocardial ischemia, we examined the effect of NG-nitro-L-arginine (a potent and selective inhibitor of EDRF release) on the coronary reactive hyperemic response in the open-chest dogs. Intracoronary infusion of NG-nitro-L-arginine at a coronary plasma concentration of 5 x 10(-5) M had no effect on hemodynamics and myocardial oxygen metabolism, but attenuated repayment of the flow debt by an average of 20.4% and 20.0% following coronary occlusion for 10 sec and 20 sec, respectively. Concomitant infusion of NG-nitro-L-arginine at the same concentration and 8-phenyltheophylline (a potent adenosine receptor blocker) at a coronary plasma concentration of 10(-5) M further attenuated flow debt repayment following 10 sec and 20 sec of coronary occlusion by 47.7 and 59.4%, respectively. These results indicate that EDRF plays a significant role in the coronary reactive hyperemic response and may cause vasodilation independently of adenosine-mediated vasodilation following coronary occlusion.

Based on our original concept, a fibroblast-inhibiting agent, chloroquine, was used against various animal tumors. Among transplanted animal tumors, the drug was most effective on relatively connective tissue-rich Bashford and Brown-Pearce tumors, as reflected by prolongation of life span, inhibition of tumor growth, inhibition of lowering of liver catalase activity, improvement of iron metabolism, increase of tumor necrosis, inhibition of connective tissue formation, and decrease of acid mucopolysaccharide. On the other hand, it was of little advantage in Ehrlich, Yoshida and MH134 tumors which contain little connective tissue, except for a decrease of the amount of ascites and ascites tumor cells in the former two tumors. These results indicate that chloroquine suppress the growth of the tumors relatively rich in connective tissue. This effect of chloroquine appears to be due to the primary attack of the stromal connective tissue of tumors being followed by the degeneration of tumor cells, though its probable anti-tumor activity by the indirect effects through its anti-inflammatory and systemic humoral activities should be taken into consideration.

For the purpose to clarify the causes of X-ray disturbances a series of experiments have been conducted on biological and biochemical properties of compound lipids extracted from normal and X-ray irradiated rabbit organs with a special reference to the P³²-labeled compound lipids uptake, inhibitory action to L cell proliferation and uncoupling of oxidative phosphorylation, and the following results have been obtained. The compound lipids (lysophosphatide rich fraction) isolated from the X-ray irradiated rabbit organ have been found to possess a strong hemolytic action and also an action to inhibit the cell proliferation as well as to accelerate the respiration of the mitochondria in the rabbit liver and spleen. It has also been proven that they act as to induce a marked swelling of mitochondria, to impede the formation of high energy phosphate as well as to act as an uncoupler of oxidative phosphorylation in vivo. In the test to see the uptake of P³²-labeled compound lipids by various organs, a marked uptake has been observed in spleen, bone marrow, and liver of both irradiated and non-irradiated groups. Further, the uptake of P³²-labeled compound lipids in the rabbits given intravenous injections of compound lipid fraction for 30 consecutive days previously has been found to be greatest in pancreas followed by bone marrow, spleen, liver in the order mentioned in male group, whereas it is greatest in spleen, followed by liver and bone marrow in the female group. With these results the discussion was conducted concerning the relation between the lipid metabolism and X-ray disturbances.

A granuloma pouch was formed on the back of rats by the original method of SELYE. Seven days when granuloma tissue reached its maximum, 35S labeled ChS, 59Fe labeled ChS-Fe, labeled ferric ammoninum citrate and colloidal 198Au were injected into the pouch and their absorption and organ distribution examined and compared with the results in the case where 59Fe labeled ferric ammoninum citrate and colloidal 198Au were injected into the gluteal muscle. 1. When 35S labeled ChS was injected into the granuloma pouch, radioactivity of the organs per gram tissue was high in the kidney, liver, bone marrow and spleen, in descending order. The maximum activity was seen 12 to 24 hours after injection, which is slow compared to the results obtained by KISHIDA in intraperitoneal and oral administration. 2. The absorption of Ch S-Fe by pouch where the iron is enveloped by the large ChS molecule, is slower than that of ferric ammonium citrate, an inorganic compound. 3. The uptake of Fe from the blood by bone marrow is larger when the increase of blood Fe ion concentration is slow, rater than when the increase is rapid. 4. When conoidal 198Au is injected into the pouch and injected into the" gluteal muscle, the 198Au is phargocytozed by the reticuloendothelial system organs, the liver showing the largest uptake among all organs. 5. In the intramuscular injection of colloidal 198Au and 59Fe labeled ferric ammonium citrate, radioactivity of pouch fluid is lower than that of blood. However, the difference between the two is less in the case of colloidal 198Au. 6. In the granuloma ponch, radioactivity of the abdominal wall proves to be greater than that of the dorsal wall.

1. For the purpose to clarify the mechanism of the revolutional changes in energy metabolism during the reticulocyte maturation the metabolisms of glucose and of the pentose moieties of acid soluble nucleotides have been observed on rabbit reticulocytes incubated in vitro under various conditions. 2. The maturation of reticulocyte proceeds by using the energy produced by aerobic glycolysis and is arrested in the glucose deficient medium, but the pentose moieties of purine nucleotide and nucleoside added exogenously serve as the energy source for reticulocyte maturation even in the absence of glucose. 3. The test on the utility efficiency of glucose and inosine as the energy source for reticulocyte maturation revealed that glucose is used more effectively than the pentose moiety of inosine under aerobic condition, which is advantageous for reticulocyte maturation, and vice versa under anaerobic condition, which is comparable to the metabolism of mature red cell. 4. From these results it has been suggested that the maturation of reticulocyte is the process of degradation of RNA and acid soluble nucleotides supported by the aerobic glycolysis, where the degradation products of RNA and acid soluble purine nucleotides provide the purine derivatives as the material for ATP synthesis (36) and the pentose moieties as energy source. 5. A possible mechanism for the superior utility of glucose to nucleoside pentose during reticulocyte maturation and vice versa in mature red cell has been discussed.

For the purpose to see how the suppression of the nucleic acid synthesis disturbs the cell specialization process the author observed the erythroid cell specialization in anemic rats by treating them with aminopterin (AP) and 5-bromouracil (BU). The observations indicate that the AP injection inhibits the mitosis of erythroblast with the acceleration of hemoglobin synthesis and the denucleation. The bromouracil administration scarcely suppressed the mitosis and the appearance of acidophilicity of erythroblast was retarded. Data indicate that the inhibition of mitosis accelerates the specialization or somatic protein synthesis of erythroblast. The acting mechanisms of the medicaments were discussed from the characteristics of these agents as the analogue of the substances related to DNA metabolism.

For the purpose of revealing the interaction between macrophages and plasma cells in relation to antibody formation and information for cell specialization, the proliferation of plasma cell by antigenic stimulation was observed in the rats whose RES had been previously injured by radiogold. The production of the circulating antibody was markedly suppressed by the pretreatment with radiogold. Histological observation revealed that the plasma cells and lymphocytes were completely obliterated and the tissues were replaced by the basophilic cells and fibroblastic cells. Lymph nodes which contained less radiogold and expected to be less in cell injury had also lost their lymphocytes, but showed a marked proliferation of plasma cells in the medullary cord and large basophilic cells in the area of lymph follicles. The data suggest that the impaired immune response will be due to the failure of the macrophages in releasing the informational substance for plasma cell specialization and for antibody formation on account of possible inability in metabolizing the ingested antigen by the injured macrophages.

With the bone marrow of anemic rats, which had received the repeated injections of phenylhydrazine once a day for three to four days, the effects of aminopterin and bromouracil on the nucleic acid metabolism of erythroblasts were observed in vivo experiment. The injection of aminopterin suppressed DNA synthesis with the lowered labeling index as observed by the incorporation of ³H-thymidine into DNA in vitro. But the grain count per cell showed the level similar to that of anemic control. RNA synthesis was not interfered by AP injections. These results indicate that AP mainly suppresses the thymidilate kinase. Bromouracil showed no such effect even on the administration of a large dose. On the basis of the data obtained from the experiment by using AP, a discussion was made on the correlation between DNA synthesis, nuclear function and the cell specialization.

1. For the settlement of carbon origin of urinary isovalthine, acetic acid-2-C14, valine-U-C14 or leucine-U-C14 was administered to rats together with isovaleric acid as an isovalthinuria inducer, and urinary isovalthine excreted was tested by autoradiography. As the results of which, it was found that these isotopic compounds were not the precursor of urinary isovalthine. Although the isovalthinuria inducing effect of isovaleric acid was fairly diminished by these isotopic compounds, urinary isovalthine was detected by paper electrophoresis. 2. Some metabolic products of these isotopic compounds were also inquired in urine and some tissues. The results were as follows: a) Acetic acid incorporated into urea, aspartate, serine, glutamate, proline, glycine, alanine, ornithine, ethanolamine, r-amino-buthyric acid (brain only), cholesterol and fatty acids. b) Valine incorporated into urinary glutamate and glycine, and tissue cholesterol and fatty acids. Valine was rapidly excreted in urine and found in a very small amount in liver digest. c) Leucine incorporated into urinary aspartate, serine, glutamate and glycine, and tissue cholesterol and fatty acids. 3. Several important problems of isovalthine studies to be elucidated were discussed.

Cb strain female mice were exposed to 800 p.p.m. of carbon tetrachloride for 3 hours by the use of newly devised gas chamber via constant current of gas. Contents of ATP, triglyceride and total lipid in the liver were measured at appropriate intervals after inhalation of carbon tetrachloride and compared to non-treated controls. And P : 0 ratio of the liver mitochondria was measured by oxymeter and morphological changes of liver mitochondria were observed by electron microscopy. The following results were obtained. 1. ATP conten t in the liver decreased slightly immediately after inhalation, rapidly decreased until 4 hours after inhalation and gradually decreased until 20 hours after inhalation. 2. Contents of total lipids increased slightly immediately after the exposure and increased gradually until 20 hours later. Contents of triglyceride in the liver increased at almost constant rate during and after the exposure. 3. P : 0 ratio of liver mitochondria did not change immediately after the exposure and gradually increased after the exposure, keeping parallel relation to decrease in ATP content in the liver. Decrease in ATP content in the liver after inhalation of carbon tetrachloride seems to be mainly due to uncoupling of oxidative phosphorylation of liver mitochondria. 4. Morphological changes of liver mitochondria were observed at 4 hours after the exposure by electron microscopy. 5. Decrease in ATP levels of the liver suggested to have a close relation to accumulation of lipid in the liver after the inhalation of carbon tetrachloride.