Okayama University Medical School

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.

A report is made on a case of liposarcoma of stomach in a 42 year old man. This is the first case of liposarcoma of stomach reported in Japan. The patient has remained asymptomatic for five years after operation.

The rats which received the repeated intra peritoneal or intravenous　injections of methyl palmitate showed a marked depressed phagocytic activity　of the RES as shown by the clearance test with radioactive iron as　well as by histological observations and a significantly suppressed antibody　formation against the challenge by BSA.　Differing from the cases of the blockade of the RES made by PVP or　radiogold, the injection of methyl palmitate did not result in any injurious　effect on the lymph follicles of lymph nodes and spleen and the plasma　cells proliferation as revealed by the histological observation. Histochemical observations of iron phagocytosis of the RES done by　Perls stain revealed that methyl palmitate　suppressed the phagocytic activity　of the Kupffer cells of the liver dramatically and also suppressed the　phagocytic activity of the sinus-lining cells in spleen to a lesser degree.　The result indicates that the injection of methyl palmitate attacks the phagocytic　function of the RES selectively and induces the reduced immune　response of the organism without giving any damages to the proliferation of immunologically com petent cells.　The fact suggests that the RES lowered in their phagocytic activity　fails to produce the informational substance for immune　response, showing　a lower level in the antibody formation even in the presence of antigen　and proliferating immunologically competent cells.

For the column chromatographic isolation of individual phospholipids from the total phospholipid mixture, silicic acid, DEAE cellulose, alumina and others, have been used as adsorbent. However, it must be emphasized that silicic acid (1, 2, 3, 4) is the most useful adsorbent for the separation of the total phospholipid mixture from each other in reasonable purity. VAN DEENEN reported that pure phosphatidyl glycerol was obtained from the lipid fraction of spinach leaves after repeated chromatography on silicic acid column (5). The phospholipid extracted from Escherichia coli B consists of abundant phosphatidyl ethanolamine (70-80 %), cardiolipin, phosphatidyl glycerol and other minor components as described in the previous paper (6). The high percentage content of phosphatidyl ethanolamine renders it difficult to separate the phospholipids by the column chromatography. Therefore, repeated chromatographies on the silicic acid column treated with sodium bicarbonate (7) and normal silicic acid column were employed for the isolation of the major components from the total phospholipid of E. coli B. Stepwise elution (4) was carried out with chloroform containing increasing proportions of methanol, and the eluent was divided into several fractions according to experience with thin-layer chromatography.

Replacement of abdominal vena cava with a fresh autogenous substitute, the segment of small intestinal submucosa, was attempted in 15 animals. Five segments were prepared from the intestine smeared with iodine tincture, and reinforced with a steel coil externally in the entire length and a steel or polyethylene ring at the anastomosis. Thoracoabdominal long implantation was done in three animals, of which one with the intestinal segment devoid of mucosa, and the other two with the submucosa. Replacement of abdominal vena cava with the submucosa taken out of the intestinal segment preserved for nine days in 1% mercurochrome solution, or 0.1 % acrinolum solution was done in one animal each. In these two a coil and two rings were also applied. Replacement of abdominal aorta with the double layer tube of a reconstructed submucosa and another very porous Tetoron gauze was done in two animals, each coupled with the abdominal vena cava replacement at the same time. Of these experiments, aorta replacements were nearly patent in both. The abdominal vena cava replacements made of the submucosa treated with iodine tincture were patent in three. The one that was preserved in acrinolum showed moderate constriction. Most of the others were also observed for a long period of time but these all occluded in spite of the initial patency which was revealed at three to seven days in cavograms, and the time of the occlusion was not determined. The internal surface of the segment of submucosa, being implanted, is covered in the first stage with the deposition of fibrin, which is subsequently organized into a fibrous layer, in the same manner as that of the synthetic graft. Another disadvantage of this substitute is its readily collapsible tendency. Infection is preventable in the experiment. The substitute seems to be useful for the replacement of aorta and for the short segment of vena cave.

Experimental replacement of inferior vena cava with crimped woven Tetoron arterial graft was performed in dogs. Bypass-graft to thoracic inferior vena cava was not successful in two animals. Total repacement of thoracic inferior vena cava was attempted in four animals, and thoracoabdominal long implantation to inferior vena cava through diaphragm behind liver, followed by excision of thoracic inferior vena cava between the anastomoses, was done in 12 animals. Of these 16 animals, the graft was patent or not occluded in nine at autopsy between the 30th and the 451st day after implantation. Similar thoracoabdminal implantation of a graft reinforced with a steel coil was made in seven animals. Two grafts were patent at autopsy after 37 and 251 days, respectively. Abdominal vena cava replacement with a graft reinforced with a coil was undertaken in three animals. Two grafts were patent at autopsy after 117 and 142 days, respectively. On the whole, long term survival without occlusion over 30 days was obtained in fifteen/twenty-eight animals. Aside from the instances of simple bypassgraft and obvious technical errors in early experiments, it was in fifteen/ eighteen, and the graft was completely patent in ten/eighteen animals. The failures within 30 days resulted mostly from either lung complications or technical errors, and the latter were remarkable in the thoracoabdominal group where the graft reinforced with coil was used, but the application of the coil was very effective in protecting the graft against the compression by the adjacent organs. Tissue reaction to Tetoron was not noticeable and to the silk thread it was very slight and seemed not to affect long term success. By the present method even the total replacement of theracic inferior vena cava can be performed safely under normothermia and thoracoabdominallong implantation to inferior vena cava is also possible with considerable success. In order to prepare a more suitable synthetic graft for vein, it requires further search for harder, lighter, more elastic and physicochemically more stable material. The fabric of venous graft should be preferably more porous and thinner than that of the arterial graft available at the present in order to make the organization within the shortest time possible.

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.

The energy source required for the amino acid incorporation into mitochondrial proteins has been investigated and comparative study has also been made on the rate of the amino acid incorporation in rat liver and rat hepatoma cell mitochondria. 1. The incorporation of amino acid into the protein in intact mitochondria of rat liver increased by about 40% on the addition of α-ketoglutarate and ADP, but no significant increase in the amino acid incorporation was observed on the addition of succinate and ADP. 2. The incorporation of amino acids into mitochondrial proteins was remarkably inhibited by the addition of respiratory inhibitors (cyanide, DNP at a high concentration). 3. The amino acid incorporation into mitochondrial proteins was scarcely or slightly inhibited by the addition of DNP at the concentration of 1×10-4M and insensitive to oligomycin (5 to 10 μg/ml). 4. The amino acid incorporation into the protein in the endogenous substrate system of the mitochondria was considerably inhibited by the addition of arsenite, and this inhibition somewhat recovered on the addition of ADP plus succinate. 5. The rate of the amino acid incorporations between rat liver and hepatoma cell mitochondria was at the same level. 6. Discussions were made on the energy source required for the amino acid incorporation into mitochondrial proteins, on the rate of protein synthesis per mitochondrion isolated from rat liver- and hepatoma cells, and on the possibilities of contamination of bacteria or microsomes and of the adsorption of amino acids onto the mitochondria.

In Anbetracht der oft schwierigen Umgrenzung des Ekg Bildes erschien es von Interesse, an einem gröβeren Personenkreis mit einem neuen Beobachtungssystem Untersuchungen unter dem Gesichtspunkt anzustellen, ob Herzgesunde charakteristische Veränderungen der Herzstromkurve aufweisen. In der vorliegenden Arbeit wird im Hinblick auf diese Fragestellung über Rhythmusstörungen im Tagesablauf berichtet, die bei 100 Herzgesunden jüngeren Erwachsenen während einer kontinuierlichen Magnetbandregistrierung von nahezu 24 Stunden Dauer gefunden wurden. AUβerdem wird das Auftreten der RhythmusstOrungen bei "Herzgesunden", mit dem 35 Patienten der "alten Gruppe", die anamnestisch und physiologisch an keiner Herzerkrankung leiden und mit 100 "Koronarleiden" verglichen. 1. Die beobachteten Rhythmusstörungen waren bei allen Gruppen .sehr ausgeprägt, d. h. jede fUr sich 55%, 100%, 97%. 2. Die am häufigsten beobachteten Abweichungen bei allen Gruppen waren Extrasystolen (ES) d. h. 49%, 100%, 97 %. 3. Häufiger fanden sich mehr supraventribuläre ES als ventrikuläre bei Herzgenuden, (31: 29 FäIle), bei der "alten Gruppe" (29: 22 FäIle), nicht dagegen bei "Koronarleiden" (67: 92 FäIle). 4. Von allen ES fanden sich die Kammer-ES am häufigsten. (29, 22, 92 FäIle). 5. Der Schrittmacherwechsel trat nur bei einigen der Herzgesunden und zwar ausschlieBlich nachts während des Schlafes auf. Diese Tatsache müβte auch seine funktionelle Genese bestätigen. 6. Vorhoffimmern und paroxysmale Tachykardien traten bei Herzgesunden nie auf. Derartige Erscheinugen können auβerordentlich bedeutungsvoll sein. 7. Sicherlich ist das Vorhandensein eines Av-Blockes 2. bzw. 3. Grades ein Hinweis auf die Notwendigkeit einer sorgfältigen und kritischen Untersuchung. 8. Wenngleich je nach den Umständen und der Tageszeit des Auftretens für die meisten Kurvenabweichungen und RhythmusstOrungen Änderungen des vegetativen Tonus von wesentlicher Bedeutung sein durften, fanden sie sich bei Personen mit erhöhrter vegetativer Labilität dennoch nicht gehäuft. Aus den Ergebnissen wird der Schluβ gezogen, daβ in der Bewertung der hier beobachteten, meist flüchtigen Veränderungen bei der Ekg-Beurteilung Zurückhaltung geboten ist, Kontroll- und Funktions-Ekg sollten häufiger als üblich zur Abklärung herangezogen werden, ehe die Diagnose einer pathologischen Abweichung gestellt wird.

For the purpose of revealing the role of the reticuloendothelial system (RES) for the antibody formation, the rats which received the repeated intraperitoneal and subcutaneous injections of a vast amount of PVP were challenged by bovine serum albumin (BSA) introducing through 2 routes of intramuscular and intravenous, and then antibody formation was observed. Blood cell count and clearance rate of radiogold were observed for the purpose of obtaining the information of blockade grade of the RES by PVP. Phagocytic activity of macrophages ingesting PVP against iron colloid were also observed in vitro. 1. A severe anemia was induced by the administration of a vast amount of PVP, 15 ml of 3% solution daily or every other day for 63 days. Histological picture indicated the suppressed erythropoiesis probably by iron deficiency or the lowered iron transporting activity of the RES, as the anemia recovered after intraperitoneal iron injections. 2. With the generalized and marked swelling of the RES, the cells in germinal center of spleen and lymph nodes were extremely swollen and lymphocytes disappeared completely, suggesting that the macrophages in germinal center play an important role in reproduction and differentiation of lymphocytes. 3. The phagocytic activity of the RES as understood from the clearance rate of radiogold was suppressed only slightly even by a heavy deposition of PVP after the repeated injections. The state of blockade or the suppressed phagocytic activity persisted for 48 hours or more after the several PVP injections. However, complete blockade of the RES or inactivation of the phagocytic activity by PVP injection was not attained. 4. A prolonged treatment of animals with PVP caused delay in the appearance of circulating antibody but the final titration reached the same level as that of control. The data suggest that the blockade of the RES by PVP induces the delay in the transmittance of the information for the antibody formation from the macrophages to the immunologically competent cells but no delay in the ingesting antigen by the macrophages.

The synthesis of nuclieic acids in the liver and the lymphoid tissues of adult mice was studied during the restitution period after the 6-day starvation. The results obtained indicate that there occurs an unexpected rapid synthesis of DNA in the hepatic parenchymal cells during the restitution period without significant increase in the total amount of DNA in the liver. Most rapid DNA-synthesis in the liver appears to occur one day after refeeding. With respect to RNA in the liver as well as to both RNA and DNA in the lymphoid tissnes, on the other hand, there is a good parallelism between the rate of their synthesis and that of increase in their amounts, without apparent dissociation between both rates as seen in the liver DNA.

1. The studies of structure and function of the plasma membranes of cancer cells is extremely important for the elucidation of specificity of phenotypes of cancer cells. In order to bring this subject to light, plasma membranes, mitochondria, microsomes and nuclei have been isolated from the AH 130 ascites carcinoma cells and rat liver cells. The electron cytochemical observations and biochemical assays of M g²+-Na+-K+-ATPase, ADPase, AMPase, and β-glycerophosphatase activities have been carried out before and after the fixation with glutaraldehyde. 2. M g²+-ATPase and Mg²+-N a +-K +-ATPase are present in the isolated plasma membranes, mitochondria and microsomes in both AH 130 cells and rat liver cells. ADPase and AMPase of the mitochondria and microsomes show far lower activities than those of the corresponding enzymes found in rat liver plasma membrane. ADPase and AMPase of AH 130 cell fraction exhibit activity much lower or zero. Generally, enzymatic activity of the AH 130 cell fraction is much lower than that of rat liver cell fraction. 3. Mg²+-Na+-K+-ATPase is completely abolished by 5% glutaraldehyde fixation while it shows less effect on Mg²+-ATPase in the plasma membrane. ADPase and AMPase activities of the mitochondria and microsomes are completely inhibited by glutaraldehyde fixation. AMPase of the plasma membrane of rat liver is completely abolished while ADPase activity is not affected in any way. 4. Only Mg²+-ATPase can be demonstrated electron cytochemically. Cytochemical reaction products of Mg²+-ATPase are located at the outer layer of the plasma membrane of the AH 130 cells and rat liver tissue. In the isolated membrane fractions it is located at the inner layer. 5. ρ-Chloromercuribenzoate has only a slight effect on Mg²+-ATPase and Mg²+-Na+-K+-ATPase activities of the rat liver membrane, while it inhibits these enzyme activities in the AH 130 cell membrane. NaF (1 mM) and NaN3 (1 mM) inactivate ADPase of the rat liver plasma memo brane. 6. In these experimental conditions, nonenzymatic hydrolysis of ATP by lead ions is not recognized. 7. It seems most reasonable to conclude that cytochemical electron microscopic demonstration of Mg²+-ATPase after fixation with glutaraldehyde may serve as the absolute marker for the plasma membrane of ascites hepatoma and liver cells.

For the purpose to reveal the characteris6cs of the synovial fluid of the chronic rheumatoid arthritis the proteins of the synovial fluid and blood serum have been analysed by employing the methods of electrophoresis, gel filtration on Sephadex G-200 column and ultracentrifugation. Waaler-Rose test and latex fixation test have also been made on each protein fraction, and the following results were obtained. 1) The total protein level of synovial fluid, which is 3/5 of that of the serum, is slightly higher than that of control. 2) Fractionation of the synovial proteins by electrophoresis revealed nearly the same protein contents in each fraction in percentage as that of comparable fraction of the serum protein, with a slight increase in γ-globulin fraction. 3) The fractionation by Sephadex column G-200 give three peaks both in serum and synovial fluid, 19 S, 7Sand 4S. 4) 19S fraction of the synovial fluid, which is mainly of γ-globulin, showed a higher level than that of the synovial fluid from the controls. 5) Rheumatoid tests gave positive reaction in the 1st peak containing 19S γ-globulin from the synovial fluid and blood serum.

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.

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.

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.

The contents of nucleic acids in rat liver and hepatoma mitochondria and the physico-chemical properties on DNA's isolated from these mitochondria were comparatively investigated. The results are briefly summarized as follows. 1. The contents of DNA and RNA per mg protein of the hepatoma cell mitochondria were about 10 and 2 to 4 times higher than those of rat liver mitochondria, respectively. 2. The λ max. and λmin. values of DNA isolated from the hepatoma mitochondria were 257 mμ and 231 mμ, respectively and those of DNA isolated from the nuclei were 259 mμ and 233 mμ, respectively, in saline-citrate, pH 7.0. 3. Three fractions of mitochondrial DNA were obtained by the sucrose density gradient and these DNA fractions corresponded, probably, to about 30 S, and 20 S and 14 S DNA's. 4. There was little difference in base compositions between nuclear and mitochondrial DNA's of the hepatoma cells. 5. The degree of hybridization between the nuclear and mitochondrial DNA's of the hepatoma cells was almost the same as that between the nuclear and nuclear DNA's of the hepatoma cells, and somewhat higher than that between the nuclear DNA of rat liver and the nuclear DNA of hepatoma cells. 6. "Highly twisted" circular, "open" circular and linear forms were observed in the DNA preparations of the hepatoma mitochondria. The average values of contour lengths of rat liver and the hepatoma DNA's observed at high frequency were 5.3 μ and 4.5 μ. 7. A discussion was made on the relation between the genetic informations of mitochondrial DNA and the formation of a mitochondrion in rat liver and the hepatoma cells.

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.

For the purpose of settling the specialization stage of erythroblast where the transcription for hemoglobin is initiated, the absorption of heme and the incorporation of tritiated uridine into RNA have been observed on the cells from the anemic rabbit after a mass red cell transfusion by which the DNA synthesis of large size precursors is suppressed and the early denucleation of erythroblasts is stimulated. In the erythroblasts obtained 24 to 72 hours after red cell transfusion a distinct absorption of heme appears first in the proerythroblast, followed by a progressive increase with the advance of the specialization. Hemoglobin synthesis is markedly stimulated after the denucleation. The incorporation of tritiated uridine into RNA is most marked in the proerythroblast and decreases with the advance of specialization stage suggesting that the mRNA synthesis for hemoglobin is initiated at the proerythroblast, continuing to the polychromatic erythroblast where. the synthesis is minimized. The volumetric observations indicate a possible denucleation at proerythroblast, but it has been revealed that the maximum RNA level of macrocytes is comparable to that of early basophilic erythroblast and its highest hemoglobin level is only that expected in the cells denucleated at late basophilic stage. From these observations it has been concluded that the transcription for hemoglobin is triggered at the initial step of erythroid cell specialization, proerythroblast, but it is insufficient for the synthesis of the expected amount of hemoglobin and is compensated or completed by the mRNA synthesis in more advanced stage of specialization.

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.