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Correlation of molecular structure with biochemical functions of the plasma membrane of the microvilli of intestinal epithelial cells has been investigated by biochemical and electron microscopic procedures. Repeating particles, measuring approximately 60 Åin diameter, were found on the surface of the microvilli membrane which had been isolated or purified from rabbit intestinal epithelial cells and negatively stained with phosphotungstic acid. These particles were proved to be inherent components of the microvillus membrane, attached to the outer surface of its trilaminar structure, and were designated as the elementary particles of the microvilli of intestinal epithelial cells. Biochemical and electron microscopic identification of these elementary particles has been carried out by isolation of the elementary particles with papain from the isolated microvillus membrane, followed by purification of the particles by chromatographies on DEAE-cellulose and Sephadex columns. The partially purified particles containing invertase and leucine aminopeptidase are similar in size and structure to those of the elementary particles in the microvillus membrane. Evidence indicates that each of the elementary particles coincide with or include an enzyme molecule such as disaccharidase or peptidase, which carry out the terminal hydrolytic digestion of carbohydrates and proteins, respectively, on the surface of the microvillus membrane. Magnesium ionactivated adenosine triphosphatase and alkaline phosphatase cannot be solubilized with papain but remains in the smooth-surface membrane after the elementary particles have been removed. Cytochemical electron microscopic observation revealed that the active site of magnesium ion-activated adenosine triphosphatase is localized predominantly in the inner surface of the trilaminar structure of the microvillus membrane.

We fractionated the red cells of Japanese acatalasemic individuals (four individuals in two families) by SAss's method and counted the number of reticulocytes and determined the catalatic activity by manometric and perborate methods on each fraction containing reticulocytes in the descending order from the upper to the lower layers. We also incubated each of these fractions at 37°C for 4, 10,24 and 48 hours, to see the catalatic activity along with the maturation of reticulocytes. Heat stability of catalatic fraction separated by DEAE column was also examined. The results of the study may briefly be summarized as follows. 1. a. There was no parallel relationship between the number of reticulocytes and the catalatic activity in Japanese acatalasemic red cells. b. There could be seen no decrease in catalatic activity while the number of reticulocytes did decrease by the incubation. 2. Heat stability of crude catalatic fraction from acatalasemic blood is approximately the same as that of crude catalase fraction from normal blood.

An attempt was made to find out the nature of catalase coritained in the red cell, especially in the ghost, For this the red cell ghost isolated were washed several times with CO2-saturated water or deionized water and the catalase activity per gram protein of the ghost was estimated. It was found that despite several washings, the catalase activity/gram protein of the ghost do not decrease as compared with the activity of the original red cell solution, indicating the presence of catalase in the ghost. In the case of hypocatalasemic blood the catalase activity in the ghost shows similar behaviors as with normal blood cells. It is assumed theoretically that there are two kinds of catalase having different affinity to the red cell ghost. Namely, one that is readily released from the ghost and the other that has a strong affinity. The affinity of hypocatalasemic blood to the ghost seems to be somewhat weaker.

The following conclusions were drawn from the above data concerning the RES function to sequestrate 51Cr-labelled heat-damaged iso-erythrocytes in mice. (l) When the hematological disorders of mice were induced, the RES of the liver and spleen reacted in the same manner. (2) The RES function of the bone marrow and liver were observed to react reversely except in the case of splenectomized mice. (3) Human gamma globulin hypersensitization and chloramphenicol administrations suppressed RES function of the bone marrow and augmented that of the liver and spleen. (4) The RES function of the bone marrow was activated after splenectomy. (5) The massive human gumma globulin administration was followed by the increased RES function of the bone marrow and by the suppressed one of the liver and spleen.

1. A cytochrome oxidase-rich submitochondrial membrane (green membrane) was obtained from beef heart mitochondria after extraction of flavoproteins, cytochrome b, Cll C, etc. by treating with deoxycholate and potassium chloride. 2. The green membrane was formed by self assembly from the membrane fragments (flat sheets), which derived from the cristae membrane of mitochondria and had essentially the same particulate structure as the green membrane. 3. The green membrane exhibited regular arrays of small particles on the surface, measuring approximately 50 to 60 A in diameter with center to center distance of about 70 A. These particles sometime were arranged in a woven structure on the surface. 4. Both the configuration of the particles and the regularity of the arrangement were influenced by detergents and temperature. 5. Green membranes as well as beef heart mitochondria and electron transfer particles commonly retained membrane-structure after sonication and exhibited higher specific activity of cytochrome oxidase than that of purified cytochrome oxidase, if the activity is calculated on the basis of heme a concentration (sec1 / 10 m,lJ.moles of heme a/3 ml). The results suggest that the active sites of cytochrome oxidase are arranged on the surface of these membranes. 6. From these results and other experimental findings, an intimate correlation between cytochrome oxidase and the particles observed on the green membranes is suggested.

<P>The mitochondrial, the microsomal, and the supernatant fractions were prepared from the cell homogenate of tumors induced by viruses, such as adenovirus type 12, SV 40, and Rous sarcoma virus, etc. and the antigenicities of these fractions were investigated. In the virus-induced tumors, there existed no antigenicity common to the mitochondrial and the microsomal fractions as in the tumors induced by chemical carcinogens, and the highest antigenicity was recognized in the mitochondrial fraction. Therefore, the properties of the tumor cell mitochondria were precisely investigated with virus-induced tumor mitochondria. 1. The mitochondria of tumors induced by viruses have clearly the specific antigenicity. 2. This specific antigenicity of virus.induced tumor mitochondria IS common to all the virus-induced tumors used in the present study. 3. This tumor mitochondria.specific antigenicity is found commonly in all the tumor mitochondria in the present experiments. 4. The specific cancer antigenicity of tumor cell mitochondria does not exist in normal organ mitochondria, but the regenerating organ mitochondria exhibit a slight antigenicity common to cancer cell mitochondria.

The regional lymph node cells of the mice sensitized with Ehrlich ascites tumor cells is known to possess a substance that shows antitumor activity on target cells (JTC-II cells). For the purpose to clarify the localization of this substance the regional lymph node cells from such sensitized mice were treated with trypsin solution of different concentrations (1.0 %, 0.2 %, and 0.01 %), and the tissue culture was carried out with JTC.II cells. As a result it was found that these lymph node cells lost antitumor activity. Next, by the differential contrifugation of these sensitized lymphocytes we obtained F1 fraction (700 g, sediment), F2 (8,500 g sediment), F3 (100,000 g sediment) and F4 (100,000 g supernatant). In the presence of each of these fractions tissue culture was conducted with JTC-II cells as target cells, and it was found that the substance with antitumor activity is contained abundantly in F2 fraction (8,500 g sediment) and F4 fraction (100,000 g supernatant). After giving due consideration to the results of these two experiments and also to the available data in the literature, we assume that the substance with antitumor activity is contained in the cell membrane component.

In the mixed tissue culture of mouse lymphocytes with addition of PHA the rate of the appearance of large and intermediate cells increases markedly, but which side of the two cell groups have reacted stronger remains obscure. In order to solve this problem, mixed cultures were conducted in such a way that only one cell group of the two would react. Namely, one cell group was exposed to C0 6).irradiation (Table 2) prior to the culture and cultured with another viable cell group (FJ test group, Table 2) to see the percentage of the appearance of large and intermediate cells. Simultaneously, the skin homograft from respective donor mouse was transplanted to each other and the survival days of each skin graft were compared. As a result it has been shown that the percentage of blastformation and the survival time of the skin transplant in each group prove to be in an inverse relation. The results of these mixed cultures indicate that the extent of blast. formation reflects significantly the difference in B-2 histocompatibility antigens.

In this review of the chemistry, biochemistry and chemical pathology of the bile pigments I am well aware that I shall ask many more questions than I am able to answer. It seems appropriate that the subject should be considered under the five headings: -Chemistry; Formation from Haem Proteins; Transport in the Blood; Conjugation and Transport to the Bile; and Changes in the Gut. I shall conclude with a brief account of the early labelled bilirubin. Because investigation of the pathological significance of the chemical changes undergone in the body is possible only if the chemical structures of the bile pigments are accurately known, my department in London has been very much concerned during the last 15 years with the chemistry of the bile pigments. The work I shall describe has been carried out in collaboration with Dr. NICHOLSON, Dr. KULCZYCKA, Dr. COLE, Dr. PETRYKA and more recently by Mr. STOLL and Miss LEMMON.

Die Autoren berichten uber die guten Behandlungsmoglichkeiten in der Praxis bei verschiedenen peripheren venosen Durchblutungsstorungen bei Erkrankungen der Bewegungsapparate, bei leichteren Fallen von arteriellen DurchblutungsstOrungen und bei einigen Fallen von Hirnblutungen mit Vasotonin und Vasotonin-forte und auch in vitro-Blutgerinnungsuntersuchungen. Bei den erwahnten Fallen kann man die therapeutische Wirkung noch mit Vasotonin-forte-Salbe gut unterstutzen. Die Autoren konnen nachweisen, daB Extr. Arnicae insbesonder in dem Vasotonin-forte die hamodynamische und die blutgerinnungshemmende Wirkung von extr. Aesculi Hippercastani gut verstairkt.

Die physikalischen Grundlagen eines Kernreaktors werden beschrieben. Als spezielles Beispiel wird Aufbau, Kontrolle und Betrieb des Siemens-Unterrichtsreaktors SUR 100 beschrieben. Dieser homogene polyathylenmoderierte Nullleistungsreaktor hat eine Leistung von nur 100 mW. Trotzdem-oder gerade deswegen-ist dieser Reaktor fur Ausbildungszwecke und als Ubungsmoglichkeit auf dem Gebiet der Reaktortheorie und der Kernenergie sehr gut geeignet, denn die Leistungsbeschrankung erlaubt eine einfache Installierung und Betriebsweise des Reaktors. Neben seiner Verwendung als Ausbildungsinstrument kann dieser Reaktor aber auch als Strahlenquelle benutzt werden. Hiermit wurden verschiedene medizinische Praparate, die auch im Strahlenschutz Verwendung finden, bestrahlt, und anschlieBend ihre Dosisrate bestimmt. AuBerdem werden noch weitere Anwendungsmoglichkeiten des Siemens-Unterrichtsreaktors SUR 100 deschrieben.

For the purpose to clarify further the residual catalase in the blood cell ghost, the ghost has been applied to Cyanogum and starch block electrophoresis and the results are briefly summarized as follows. 1. It has been demonstrated that after Cyanogum electrophoresis of the ghost after several washings, bubbling due to enzymatic reaction of catalase occurs near the points of origin, when the plate is immersed in hydrogen peroxide solution and also it has been proved the presence of catalase so firmly bound to the ghost that it is hardly moved by the electro phoresis. Even with the ghost of hypocatlasemia there can be detected catalase which is likewise hardly eluted from the ghost. 2. In the estimation of catalase activities of each fraction from red cell ghost by starch block electrophoresis there can be detected catalase near the point of origin, that is not eluted by the electrophoresis, and the activity of which corresponds to about 0.1 % of the total red cell catalase activity.

The present investigation was carried out to see effects of muscle cornin, an alcoholic fiaction of boiling-water extract from rabbit skeletal muscle, on the nucleic acid synthesis in the early development of Pseudocentrotus depressus. In this study, the author used the assay method of our own device by which we can estimate the incorporation into whole cell simultaneously that into nucleic acid fraction, with one and the same specimen. The results of the observations are briefly summarized as follows. 1) Cornin accelerated the incorporation of 3H-uridine into whole cell by 10-20 %. 3H-thymine, 3H-thymidine and 3H-uracil all inhibited such incorporation. 2) As to the incorporation into the RNA, it was retarded in the course of phosphorylation at the synthetic stage. 3) In the incorporation into DNA, since the incorporation is inhibited by about 2/3 at the synthetic stage, it seems that the polymerization is inhibited. 4) This inhibition of the DNA synthesis was also substantiated by the autoradiography with tritiated thymidine. Some coments were made on the operation of the nucleic acid synthesis, the specific protein structure during the early development of sea urchin egg, and effects of cornin on these as well as on the other intrinsic substances.

Compound 48/80, sinomenine, tween 20 and polyvinylpyrrolidone (PVP) were injected intravenously to dogs, in doses producing similar degree of profound hypotension, and changes in the plasma histamine content and coagulation time were followed on the blood from the femoral artery. After the injection of 48/80 or sinomenine plasma histamine rose rapidly and markedly, attaining its maximum within 2 minutes, but the increase was rather of a short duration. In contrast, after the injection of tween 20 or PVP a less marked increase in plasma histamine developed more slowly, but lasted longer. The blood coagulation time was prolonged in all the cases injected with 48/80, and occasionally with sinomenine. Both beginning and recovery of the prolongation of blood coagulation time were sluggish as compared with the changes of plasma histamine. Tween 20 and PVP did not induce any detectable change of the blood coagulation time. These data were discussed with reference to the sites of action of different histamine releasers.

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.

1. It has been found that mouse lymph.node cells, even destroyed by sonication with 20 KC supersonicator, maintain sufficient antigenicity both in vitro and in vivo. 2. When such sonicated cell homogenate is cultured with live lymph-node cells, there can be observed blastformation and the peak of the rate of the blastformation is seen at culture hour 48. 3. When PHA (phytohemagglutinin)-M is added to such mixed cultures, the blastformation is enhanced. 4. When mixed cultures of mouse lymph-node cells are conducted by using such one-way stimulation method in various combinations, the rate of blastformation can tell quite accurately the differences in H-2 antigens of mice. 5. In the experiment using F1 hybrid mice and the parents, it has been demonstrated that the rate of blastformation in mixed cultures of the present experiments shows a direct correlation to the rate of blast formation in mixed cultures of live lymph node cells, whlie it is an inverse proportion to the survival time of the skin transplant. 6. Differences in the transplanation antigens said to be located on sex chromosomes cannot be distinguished by this one.way stimulation method.

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.

1. Mitochondria isolated from human liver, hepatoma and gastric cancer contain DNA. The DNA content per mitochondrial protein is about ten times as much in cancer as in normal liver. 2. Human liver, hepatoma and gastric cancer contain circular DNA molecules in their mitochondria. Circular DNAs from normal liver and cancer mitochondria are mostly about 5 μ long, and the frequency of circular DNAs of multiple or shorter length is higher in cancer mitochondrial DNA. The outline of the present paper was presented at the 26th Congress of Japanese Cancer Association (1967) (52, 53).

Five (21 per cent) out of 24 mongrel dogs were found to be refractory to compound 48/80 and also to sinomenine (cross-tolerance). These nonreactor dogs responded normally to PVP and tween 20 and showed normal sensitivity to histamine. The incidence was similar in both sexes. Mechanisms of this type refractoriness were discussed.

As a step towards the elimination of Japanese encephalitis virus in natural surroundings, we inoculated pigs, rabbits and chicks with inactivated Japanese encephalitis vaccine supplemented with complete or incomplete Freund's adjuvant twice at one-week interval. Subsequently, we compared HI antibody titers of the groups inoculated with vaccine containing complete Freund's adjuvant (pigs, rabbits, chicks), of the group inoculated with vaccine containing incomplete adjuvant (rabbits), ar;d of the groups inoculated with vaccine containing no adjuvant (pigs, rabbits, chicks), and also observations on changes in the antibody titers due to natural infection. In a certain portion of these animals neutralizing antibody titers were also determined. The results of this study are briefly summarized as follows. 1. In the groups of pigs and rabbits inoculated with vaccine containing complete Freund's adjuvant, titers of HI antibody and neutralizing antibody were higher than those inoculated with vaccine containing no adjuvant and their high titers persisted. Further, in the group of chicks inoculated with inactivated Japanese encephalitis vaccine containing complete Freund's adjuvant, HI antibody titers were higher and persistent as compared with the antibody titers in the chicks inoculated with inactivated Japanese encephalitis vaccine alone. 2. In the rabbits inoculated with inactivated Japanese encephalitis vaccine contammg incomplete adjuvant, HI antibody titers were lower than in those receiving the vaccine with complete adjuvant, but it has been demonstrated clearly that vaccination of inactivated Japanese encephalitis vaccine supplemented with incomplete adjuvant brings about less sideeffects. Hence such a method of vaccination can be applied as the vaccination with least side-effects. 3. With respect to natural infection of swine, on August 27 when the pigs were thought to have been infected, there was observed a rise in antibody titers. And on being infected with Japanese encephalitis, the antibodies formed in those pigs inoculated with inactivated Japanese ence- phalitis vaccine with or without complete adjuvant proved to be all 2-ME resistant type, whereas the antibodies produced in the control groups not receiving such a vaccination were 2-ME sensitive antibody.