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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.

For the purpose of revealing whether AMD inhibits the RNA synthesis of erythroblasts in an effective dose in vivo to eradicate erythroid cells in rabbit bone marrow, the author observed the RNA synthesis by H3-uridine incorporation in vitro and RNA level on the cells from the anemic animals taken at a certain period after a single injection of AMD in a small dose of 50 and 100μg/kg body weight. The data revealed that by such a small dose of injection of AMD the RNA synthesis of erythroid precursors, early basophilic and proerythroblast stages, was successfully suppressed without any suppressing effect on the RNA synthesis of erythroblasts in the later stages of specialization, indicating that there are at least two kinds of RNA synthesis: one seen mainly in the earlier stages of specialization and the other one seen mainly in the later stages, and they can be distinguished from each other by the AMD sensitivity.

1. After the centrifugation of sonicated heavy beef heart mitochondria at 75, 000 × g for 10 minutes, the supernatant was centrifuged at 144, 000 × g for 30 minutes. The residue was revealed being composed of vesicular inner membrane fragments (ETPH), about 600 to 1000 Å. in diameter, showing a morphological homogeneity and a high capacity of oxidative phosphorylation. 2. The Pia ratio of the ETPH in the presence of succinate and of NADH2 was 1.68 and 2.54, respectively, and the corrected Pia value for O2 gas equilibrium was 1. 01 and 1.40, respectively. 3. The capacity of oxidative phosphorylation in ETPH fraction was parallel to the activity of the oligomycin. sensitive ATPase in these fractions. 4. The P/0 ratio of ETPH was decreased to about 50 % by hypotonic treatment. The decrease of P/0 ratio was restored to the level of about 90 % by incubating the ETPH with ATP and BSA. In the instance where the P/0 ratio was low level in the hypotonic medium, the surface structure of ETPH was observed as a swollen form and the head pieces of the elementary particles were clearly observed in contrast to the solid surface structure of ETPH in the isotonic medium. 5. The P/0 ratio of ETPH was decreased to about 60 % by relatively severe sonication, and after separating the residue from the supernatant, that of the residue decreased further to about 40 %. The P/0 ratio of the residue was restored to the level before the separation on the addition of the supernatant containing oligomycin-insensitive ATPase. 6. A discussion was made on the correlation between the surface structure and the activities at terminal phosphorylation step of ETPH after the simple physico-chemical treatment.

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.

For the purpose to confirm the localization of DNA in chloroplasts and mitochondria, the cells of Spinacia oleracea fixed with glutaraldehyde-Os04 were observed by electron microscope with or without DNase treatment. "DNA fibril complexes" have always been found in the electron-transparent regions of the chloroplasts and mitochondria of the cells receiving no DNase treatment. By treating with DNase, the DNA fibril complexes of these organellae are reduced considerably in their density, leaving only faintly visible ghostlike structure or having completely disappeared. These observations confirm that the DNA fibril complexes in chloroplasts and mitochondria as demonstrated by glutaraldehyde-OsO4 fixation are the DNAcontaining structures similar to those found by formalin or buffered OsO4 fixation, and suggest that it will have only a small amount of the material other than DNA distinct from the case of DNA in the nucleus.

The authors give an account of the important developments in blood coagulation knowledge from the times of Malpighi and Moravitz to data. The article is followed by original tables providing a general and comprehensive view on blood coagulation, hemorrhagic syndromes and fibrinolysis.

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 liver cells obtained from a calf have been cultured continuously for 257 days in total at present (May 31, 1967). The primary culture was maintained in rotatory culture for about 2 months with gradual and continuous cell proliferation. The two original strains, LD-BS20 and LD-CS20, have been maintained in static culture since 4th subcultivation. Three substrains, LD-BS10, YLE-BS20 and LD-CS10, were derived from the original strains. Two kinds of appropriate media, in which the cells could be subcultured with trypsin without severe damages and maintained with some characteristic functions of liver cells, were reported. The one consisted of 20 per cent bovine serum, 0.4 per cent lactalbumin hydrolysate and saline D, and the other was added with 0.08 per cent yeast extract to the above mentioned medium. Calf serum examined was not so effective as bovine serum for cell proliferation. Morphologically, the cultured cells resembled parenchymal liver cells quite well. The cells spread wide with abundant pale staining cytoplasm and their large nuclei, oval or round, generally contained one to several nucleoli. The cells as well as the nuclei varied considerably in size, some being two to four times larger than others. Binuclear, trinuclear or polynuclear cells were also observed. No silver impregnated fiber was detected among the epithelial cells. Two attempts to characterize cell types in culture were made. First, the presence of glycogen was tested with PAS reaction and saliva digestion procedure. Secondly, the albumin formation in cultured liver cells was examined with the fluorescent antibody technique. The fact that both albumin and glycogen were observed in the cells suggests strongly that there is a possibility of the continuous cultivation of liver cells by the present method, and by these procedures it seems possible to identify functionally the cultured cells with the parenchymal liver cells.

A case of the so-called adenoameloblastoma developed in the right maxillary sinus of a 10-y-old girl was reported. The histological features of this tumor were discussed in detail. In the twenty cases of adenoameloblastoma, including the present case, reported in Japan up to the present, some statistic investigations have been made in regard to the clinical aspects.

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.

For the purpose to get the information about the control mechanism of erythropoiesis in bone marrow the author introduced a mass of homologous red cells into anemic animal and observed how the bone marrow cells and circulating blood react against the prompt normalization of the anemic condition. After the red cell transfusion which was enough to restore the anemia promptly the red cell number in the circulating blood continued to increase until 72 hours after the transfusion, reaching an extremely high level in both red cell number and hemoglobin contents. Mitotic index and the DNA synthesis as observed by tritiated thymidine incorporation into DNA proved no actual change even 24 hours after the red cell transfusion, though a marked decrease in labeling index was found in large size precursors. Histologic picture revealed the proliferation of reticulum cells. 48 to 72 hours after the red cell transfusion both mitotic index and DNA synthesis of erythroblasts have largely retarded in all series of specialization with the decreased appearance of the erythroblasts in bone marrow sections. The measurements of red cell size and the RNA contents of erythroblasts and reticulocytes proved the accelerated denucleation at the early stage of erythroid cell specialization, as early as basophilic stage resulting in a marked macrocytosis.

This new nosologic entity known as "Nasio irritable digestive tract", is defined as the reversible, functional neuromyosecreory trouble of the whole or a segment of the digestive tract, that alternates with periods of health with an irregular and long evolution, influenced particularly by psychical factors, and that develops in neurovegetative distony constitutions.

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.

A case of malignant melanoma with metastases mainly to the liver and the right ilium was treated with a gluconeogenic diet. The carbohydrate content of the diet was finally reduced to 5∼10 g per day and the remaining calories were derived from protein and fat. Increased blood citrate and NEFA concentrations, increased ketone body formation and the maintenance of a reasonable level of blood sugar confirmed the attainment of a gluconeogenic metabolic state. Definite improvements in size of a hepatic tumor, serum alkaline phosphatase activity and the general condition were observed transient1y during the dietary therapy. Growth of the tumor resumed despite the continued gluconeogenic therapy, and the patient died of cardiac failure. Concentrations of gluconeogenic enzymes, fructose-1, 6-diphosphatase, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, were all found to be very low in the tumor tissue as expected.

a) A modified procedure of the WIDNELL and TATA8 method yields rat liver nuclei manifesting a high degree of purity and activity. b) These nuclei contain a nucleoside-dependent phosphorylating activity that is readily released and apparently unrelated to either glycolysis or respiration. c) The main incorporation of the 32Pi is into ribose-I-phosphate; nucleoside phosphorylase activity satisfactorily accounts for the observed purine nucleoside stimulation of the nuclear phosphorus metabolism.

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.