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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 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 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).

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.

The amygdalofugal fibers were studied III the cat with the silver method of NAUTA-GYGAX. 1. The amygdalofugal fibers are distributed by way of the stria terminalis, the longitudinal association bundle, the inferior thalamic peduncle, and the medial forebrain bundle. 2. The amygdalofugal fibers running through the longitudinal association bundle arise in the lateral principal, intermediate principal nuclei and the lateral and possibly intermediate parts of the periamygdaloid cortex, and terminate in the lateral preoptic nucleus, the bed nucleus of the anterior commissure, the olfactory tubercle, the nucleus of the diagonal band of Broca, the nucleus accumbens, the medial and posterior septal nuclei and the basal part of the head of the caudate nucleus. In addition, there are scattered fibers coursing along the longitudinal association bundle proper. These fibers may have a widespread origin from the amygdaloid complex. The longitudinal association bundle contributes no fibers to the medial forebrain bundle. 3. The fibers, originating from the lateral principal, intermediate principal and medial principal nuclei, join the medial forebrain bundle to distribute widely in the lateral hypothalamic nucleus. A few fibers are seen to reach the ventromedial hypothalamic nucleus, and are considered to arise in the medial principal nucleus. 4. By way of the inferior thalamic peduncle some fibers from the amygdaloid complex course dorsally into the medial part of the dorsomedial thalamic nucleus at its caudal levels. They may arise widely from the amygdaloid complex. A few of them extend farther dorsally to reach the lateral habenular nucleus and the parataenial nucleus. They probably originate from the lateral principal nucleus. 5. The fibers forming the stria terminalis originate from the medial principal nucleus, the medial nucleus, the periamygdaloid cortex and the cortical nucleus, and are distributed in the bed nucleus of the stria terminalis and the lateral preoptic nucleus (preoptic component), as well as the medial preoptic nucleus, the anterior hypothalamic nucleus and the ventromedial hypothalamic nucleus (supracommissural component). The cortical nucleus, particularly its caudal part, and possibly the medial part of the periamygdaloid cortex are regarded as the main sources of the stria terminalis fibers ending in the hypothalamic region. The intermediate principal and lateral principal nuclei do not appear to contribute fibers to the stria terminalis. 6. The ventromedial hypothalamic nucleus receives amygdalofugal fibers both from the medial principal nucleus by way of the medial forebrain bundle, and from the cortical nucleus via the stria terminalis. 7. In addition to intrinsic internuclear fibers within the amygdaloid complex, some of the fibers from the complex are distributed to the ventralmost part of the putamen, the medial part of the claustrum, the periamygdaloid cortex, the prepiriform area and the anterior amygdaloid area, but do not reach the hippocampus.

A series of experiments was conducted to study the base composition of DNA in AVl2-induced tumor and host cells by paper chromatography, and it was found that DNA per cent. guanine-cystosine contents were around 42 % in both of them. The base composition of DNA of AV12 itself differs considerably from that of AVl2-induced tumor cells, while the DNA of tumor cells shows the property similar to that of host cell DNA. The genetical relationship among virus, host cells and tumor cells was discussed.

l) The submitochondrial particle system can synthesize ATP in the early phase (220 seconds after the accition of ADP) in the presence of sodium succinate and Pi, in spite of the absence of the hexokinase-glucose system, and this phosphorylation is inhibited by oligomycin. 2) The submitochondrial particle system can synthesize ATP by the base-acid transition (proton pulse) only in the presence of ADP and Pi, in spite of the absence of oxidizing substrates and the hexokinase-glucose system, and this phosphorylation is dependent on the span of pH change, and is inhibited by oligomycin and 2, 4-dinitrophenol. 3) The role of the proton vector in the oxidative phosphorylation and the proton ejection was discussed from the stand point of a new hypothesis.

Among various photosensitizing dyes, 4, 4'-dimethyl 3, 3'-di-n-heptyl-8- {2-(4-methyl-3-n-heptylthiazole) }-2, 2'-dicarbocyanin diiodide (abb. NK19), even in an extremely low concentration, is known to inhibit the proliferation of bacteria and tissue culture cells (1, 2, 3). With respect to the mechanism of such inhibitory action no other property of this NKl9 is known except that it has a marked adsorptive property to protein (4). As a step toward the elucidation of the mode of biological effect, the author studied the effect of NK19 on the energy transfer reaction of Irat liver mitochondria, followed by comparison with the mode of actions of various other inhibitors of the oxidative phosphorylation (5). NK19. NKl9 can be prepared by letting 2, 4-dimethylthiazole heptyliodide react with ethylorthoformate in anhydrous acetic acid. We used NKI9, a product of Nihon Kanko Shikiso Research Laboratories. The molecular structure is as in the following and in its MeOH state it has maximum absorbancy at 590 m,a. For the use in experiment it was made into 1 mg/ml of MeOH and was stored in the dark until used.

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.