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With a constant gas-exposure chamber newly devised, the author had Cb mice (females weighing 16.0 ± 1.5 g) inhale 600 ppm (in average) of 1, 1, 2, 2-tetrachloroethane for 3 hours. Then, the total Iipid, triglyceride and ATP levels in the liver were estimated before, immediately after, 4 hours and 8 hours after the exposure. The results of the observations are briefly summarized as follows: 1. It has been demonstrated by the chemical quantitative analyses of total lipid and others that the exposure to 1, 1,2, 2-tetrachloroethane induces fatty liver in mice. 2. Both total lipid and triglyceride levels increased almost linearly from the time of exposure up to 8 hours later. The ratio, triglyceride: total lipid, increased with lapse of time after the exposure, and of the lipid components, the increase of triglyceride was marked. 3. The hepatic ATP level decreased almost linearly from the time of exposure to 8 hours later. The value, total lipid × ATP, hardly differed from that of the control even after the exposure, and there was observed a parallel relationship between the rate of increase in total lipid level and the rate of decrease in the hepatic ATP level. 4. The intensity of hepatotoxicity of 1, 1, 2, 2-tetrachloroethane proved to be practically the same at that of carbon tetrachloride.

1) In order to study the molecular structure and electron transfer activities of mitochondrial inner membrane, dissolution and reconstitution of membranous structure and function of the inner membrane of beef heart mitochondria were carried out. 2) The inner membrane of mitochondria could be dissolved into some unit of particles 70-140 Å in diameter by the treatment with bile salts at the concentration 0.5 mg of deoxycholate per mg of protein, 0.5 mg of cholate per mg of protein and 74.5 mg of crystalline potassium chloride per ml of the suspension. 3) The dissolved unit particles readily reaggregated into a vesicular membrane simultaneously restoring over-all electron transfer activities by the removal of bile salts with dilution of the suspension.4) Isolated electron transfer unit particle fraction contammg all components of the electron transfer chain but no structural protein were soluble in aqueous solution due to some residual bile salts used in the preparation. The removal of bile salts by dilution led the dispersed particles to aggregate into membrane and restore their over-all enzymatic activities. 5) From these results and the results of the reconstitution of membrane from purified complexes as described in the previous paper, it may be concluded as follows: The mitochondrial inner membrane may consist of several kinds of repeating unit particles conjugating each other with adjacent particles. It is necessary for over·all enzymatic activities that some unit components aggregate into a single vesicular membrane. Structural proteins may play an important role in the constitution of the membranous structure and in the over-all enzymatic activities.

In the present experiments attempts were made to identify semen from various specimens such as the semen itself, spots of semen on clothes, putrefied semen or semen contaminated with blood, menstrual blood, vaginal fluid, according to the techniques of LEVONEN. As the result it has been clarified that in every instance it is possible to isolate and detect the spots of choline by spraying Dragehdorff's reagent.

For the purpose to clarify the mechanism of phagocytosis or pinocytosis, the observations on the tumor ascites, including the macrophages as well as the tumor cells, were carried out by incubating with the iron colloid with or without pretreatment by several inhibibitors of glycolysis, oxidative phosphorylation and respiration, or under hypotonic or cold environments. The results have demonstrated that there are three steps in the phagocytosis. The first step is the adhesion of the substance to the cell surface, which is not an energy-requiring process. The second step is the engulfing which proceeds by using the energy supplied by glycolysis. The third is the accumulation of the substance into the vesicles through the canaliculi connecting the cell surface with the vesicles. The discussion was made on the existence of the active site on the cell surface to which the substance can be adhered, and the accumulation mechanism of the material into the phagocytic vesicles by the membrane flow, the flowing movement of the outer lipid layer of a unit membrane through the canaliculi which connect the cell surface to the phagocytic vesicles.

The growth of JTC-11 cell line which was established from Ehrlich ascites carcinoma in vitro was inhibited by the addition of 2 per cent guinea pig serum to the control medium composed of 10 per cent bovine serum, 0.4 per cent lactalbumin hydrolysate and saline D. The concentration of guinea pig serum could be reduced to 0.02 per cent (lOγ of guinea pig serum protein/ml) with positive result, but 0.002 per cent guinea pig serum did not inhibit the growth at all. The inhibitory effect was not abolished by heating at 56°C, 66°C, and 70°C for 30 min but it was completely lost by heating at 100°C for 30 min. The inhibitory factor was undialyzable, and was inactivated with the treatment of 1mM HgCl2- Morphologically, the cells exposed to guinea pig serum showed pycnotic changes of the nuclei, accompanied by the formation of fine vacuole-like particles in the cytoplasm. Electron microscopic study revealed poor development of endoplasmic reticulum. There were more multivesicular bodies and large vacuoles with amorphous content in the cytoplasm of the damaged cells. The DNA synthesis in these cells was remarkably disturbed by 2 per cent guinea pig serum.

1. In the present experiments, Ehrlich ascites carcinoma (K-tsrain), JTC-11, and C3H mouse mammary tumor (A-strain) were used to study the inhibitory effects of two kinds of comins, crude muscle cornin and crude intestine comin. 2. Daily intraperitoneal administrations of both comins had shown a marked inhibitory effect on the Ehrlich ascites carcinoma. 3. Intestine comin was more effective on the inhibition of the growth of the Ehrlich ascites carcinoma than muscle cornin when administered intraperitoneally. 4. Daily subcutaneous adminstrations of muscle comin had no effect, but doses of 10 mg/mouse/day or 20 mg/mouse/day of intestine cornin had a slight or moderate inhibitory effect on the Ehrlich ascites carcinoma. 5. Intestine comin had an inhibitory effect on the growth of JTC-ll cells in vitro, and made the tumor cells to undergo morphological changes during incubation. 6. Daily intraperitoneal administrations of muscle comin had hardly any effect on the C3H mouse mammary tumor, but intestine comin was evidently effective in male. 7. Intraperitoneal administrations of intestine comin proved to be hardly effective on the C3H mouse mammary tumor, but only in the dose of 30 mg/ mouse/day, it had a moderate inhibitory effect in female. 8. Daily subcutaneous administrations of muscle comin had no effect on the C3H mouse mammary tumor, but intestine comin had a slight effect in male. 9. Muscle cornin had a slight or moderate effect on the C3H mouse mammary tumor, but intestine cornin was hardly effective in female when administered subcutaneously. 10. Repeated intraperitoneal administrations in doses of 30 mg/mouse/day of muscle comin produced intoxication in the treated mice. 11. In general, it seems that intestine comin is more effective on the inhibition of tumor growth than muscle comin.

In the immunofluorescent study it has been revealed that rabbit sera immunized with transformed cells induced by SV-40 DNA, produce circulating antibody capable of re:lcting with intranuclear antigens synthesized by SV-40 complyte virus transforming process, In addition, the result confirmed that SV-40 DNA replicates DNA-containing viruses in the host cell and that also the genome coding for the synthesis of SV-40 tumor antigen is resposible for viral DNA.

For the purpose of revealing the molecular organization of the mitochondrial membrane the authors attempted to clarify the fine structure of reduced coenzyme Q-cytochrome c reductase and also studied how the CoQH2-cyt. c reductase is arranged in the mitochondrial membrane by systematic analyses of fractions from the purification process of CoQH2-cyt. c reductase. 1. Purified CoQH2-cyt. c reductase contained high concentration of cyt. b (9.5 mμmoles per mg protein) and cyt. Cl (4.5 mμmoles per mg protein), and was almost free from cyt. c, a, flavoproteins, primary dehydrogenases and ATPase. The enzyme complex also showed a high specific activity (48 μmoles of cyt. c reduced per mg protein per min at 30°). 2. CoQH2-cyt. c reductase was composed of particles of about 120 Å in diameter with irregular form, some time exhibiting electron opaque cores. In the loose aggregates of the particles, the size of each particle was about 95 Å in diameter. 3. An intimate correlation was observed between the particles of CoQH2cyt. c reductase and those on the surface of the NADH-cyt. c reductase fraction. 4. Regular arrays of uniform particles (about 82 Å in diameter with a center to center distance of about 100 Å) were observed on the surface of the submitochondrial membrane (brown membrane) obtained from beef heart mitochondria by treatment with deoxycholate (0.1 mg / mg protein) and KCl (72 g/l). The correlation between these particles and CoQH2-cyt. c reductase was discussed.

As a link in the series of studies on tumor specific immunity an attempt was made to clarify specificity if any, in aggregation of sensitized lymph-node cells on target cell in vitro. For this purpose sensitized regional lymph-node cells from isologous CsH mouse transplanted with A cells derived from CaH mouse mammary cancer were incubated with M cells derived from mammary cancer of homologous Cb mouse and HeLa-Ss cells as with A cells. The results are briefly summarized in the following. These sensitized regional lymph-node cells (A-L) inhibited the proliferation of A cells and M cells in tissue culture. When the interaction between the sensitized lymph-node cells and the terget cells was pursued over a long period by cinematography, these lymph-node cells became attached to the target cell by 6-to 12-hour culture in aggregation of rosette form, and by 30 hours some of the target cells were seen to undergo lysis. However, when these sensitized lymph-node cells were cultured with heterologous HeLa-S3 cells (derived from human uterine cancer), no such phenomena were observed. In the case with untreated normal lymph-node cells (control) there could be hardly observed any inhibitory effect on target cells. When the number of the target cells on which the lymph-node cells became attached was counted along with lapse of time, it was more numerous in the case of A and M cells but only a few in the case of HeLa-S3 cells. It seems that most of the sensitized lymph-node cells that inhibit the growth of the target cells become attached and aggregated fairly specifically onto the target cells.

The compositions of nitrogen pools of ox liver, bladder bile, kidney and lung were analyzed with an especial bearing on their minor components, and some distinctive features of these tissues were described. DCEC and CMC were found in ox liver and kidney. Liver was low in free arginine and lysine, but high in ornithine, ethanolamine, and glutathione. Glycine was only a predominant amino acid in ox bile. All amino acids were contained moderately in kidney, but glutathione content was low. The concentrations of arginine and lysine were relatively high in lung.

Concentrations of ampholytes in the nitrogen pool of ox ocular tissues and nervous tissues were analyzed systematically by an automatic amino acid analyzer with a special reference to their minor components. DCEC was found in lens and also in nervous tissues. Ophthalmic acid was found in lens (highest), in retina (moderate), and in vitreous humor and spinal cord (trace). Glutathione content was extremely high in lens, and moderate in nervous tissues, retina and cornea. Carnosine content was moderate in cornea and in retina, but hemocarnosine may be rather high in nervous tissues. Anserine-like compound was found only in spinal cord, but free 1- and 3-methylhistidine were detected in most ocular tissues. Ethanolamine and γ-aminobutyric acid were high in retina and their concentrations were comparable to those of nervous tissues.

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.

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.

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.

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.

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.