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Changes of H+ gradient at various energy states of mitochondria were studied. There was a close relation between the extent of H+ gradient and the level of ATP formation; the former decreased as a result of ATP synthesis but was not completely abolished. A partial depression of H+ gradient was also observed in the presence of uncouplers of oxidative phosphorylation. The H+ gradient seemed to be more closely related to the ion translocation than ATP formation. In the presence of Ca++ the energy of H+ gradient was utilized in translocating Ca++ rather than synthesizing ATP. These findings further substantiate the chemiosmotic theory of MITCHELL on mitochondrial electron and energy transfer.

Two cases of megaloblastic anemia associated with anticonvulsant drugs were studied. Both cases were inpatients of psychiatric hospitals and had quite inadequate intakes of food. The former had lung tuber. culosis and the second had febril illness before the manifestation of anemia. Multiple examinations including bone marrow smears, serum iron levels, vitamin B12 levels, estimation of urinary formiminoglutamic acid after histidine loading and folic acid tolerance test revealed that this anemia was due to folic acid deficiency. Complete hematological responses were ob3erved with injection of folic acid. Retrospective nutritional study on the second case was carried out. The study revealed that folic acid content of the diet of this hospital was 152 fl.g of free folate and 522 fl.g of total folate. The folic acid mtake of the patient was about 80 /1.g of free folate and 280 l1.g of total folate daily during a month before the manifestation of megaloblatic anemia. Importance of additional factors for the development of megaloblastic anemia in patients receiving anticonvulsants was discussed and it was concluded that most important factor was nutritional deficiency of folic acid.

Scanning electron microscope (SEM) observations were conducted mainly on the in vitro interaction state between ]TC-II cells derived from Ehrlich ascites tumor and regional lymph node cells obtained from the mice I0 days after transplantation of Ehrlich ascites tumor. Cells cultured on the cover glass were fixed with glutaraldehyde, dehydrated with graded acetone solution, and covered with carbon and gold, were observed by SEM. The results may be briefly summarized as follows. On the surface of ]TC.II cells themselves are seen numerous fine microvilli projecting out regularly at right angle to the cell surface, which become attached to the glass, and there can be observed vacuoles in the cytoplasm. Such microvilli are lacking at the tip of the pseudopodial projection. The lymph node cells aggregated to ]TC-II cell are lymphocytes of small or intermediate size, and the pattern of aggregation varies: some lymphocytes effect an intimate contact with the surface of target cell by their cellular projections; the contact is achieved by interdigitation of microvilli between lymphocyte and target cell; b:>th cells form a bridge connection with a simple projection from each; or the two cells make a broad surface.to.surface contact. It is not possible to differentiate sensitized lymphocyte from nonsensitized one, their cell shape is spherical with rough surface and some cells show hole on the surface.

For the purpose to confirm whether carotene is converted into vitamin A mainly in the intestine, fluorescence microscope observations as well as the fluorescence microphotometry for the estimation of fading state of the fluorescence were carried out on the fresh sections of the intestine, lung and liver of rats after oral administration of 11-carotene dissolved in oil, and suspended in water, and vitamin A in sesame oil as control. Yellowsih-green fluorescence of carotene is similar to that of vitamin A in color but the fluorescence of vitamin A fades away very rapidly within one minute while that of carotene does not fade or fade more slowly than that of vitamin A. Observations have revealed that, contrary to expectation, the administered carotene is not so readily converted into vitamin A in the intestinal mucosa, but after passing through the intestine without conversion to vitamin A, it is transported to the mesenteric lymph vessels, portal vein, and reaches the liver. In the liver, carotene appears as fatty droplets or micronized particles in the parenchymal cell. The conversion of carotene into vitamin A could not be observed in the intestine, liver and lung in the observations made one hour after the oral adminstration of carotene. Hower, it seems that carotene dissolved in minute fat droplets may be converted into vitamin A at water phase in tissues, after dissolution of carotene in fat and micronization of the fat droplets.

Bovine adenovirus type 3.induced tumor was transplanted subcutaneously through 25 generations in hamsters. Histological character of the tumors showed no basic differences from the primary one through all generations. There appeared metastatic foci in the lung, liver, kidney and lymph nodes in some animals.

Partially separated double-stranded RNA from purified Rous sarcoma virus, Schmidt.Ruppin strain, was observed by electron microscopy utilizing 8.M urea and protein monolayer technique. Furthermore, viruses in pair were frequently and viruses with two nuc1eoids were occasionally observed in ultrathin. sectioned specimens of chick cells transformed by RSV. From these results taking other reports in consideration, a possible mechanism of RNA replication in Rous sarcoma virus is proposed.

Electron microscopy of the body wall of Opisthorchis viverrini shows the integument which is connected to the epidermal cell with fine protoplasmic tubules, to form a syncytium, as in Clonorchis sinensis and other trematodes. Vacuole-like secretory granules are distributed in the matrix of the integument, and mitochondria are arranged at the proximal outer surface of the integument. The crystalline inclusions are observed in the perinucleus of some epidermal cells.

In vitro cell transformation of human embryonic cells could be induced by DNA extracted from virions of SV 40 purified by density gradient centrifugation. The result shows clearly that cell transformation is in· duced by incorporation ofa part of viral DNA into the genome. In addition, for the purpose of clarifing the biological differences between the normal and transformant, the alteration of the cell membrane structures of transformants was observed from the mechanism of phagocytosis. The iron colloid particles are taken up by normal diploid fibroblasts but not by the human and hamster transformants. This fact suggests a difference in the molecular arrangement of the cell membranes between the normal and transformants. In the presence of histones, however, the transformants phagocytize the colloid particles very actively. The results show cell membranes of transformants are altered in the molecular structure responsible for the surface charge.

Japan was the first Asian country to introduce social insurance measures and it has expanded them during the last few decades. The first social insurance law was passed in 1922, dealing with worker's health insurance, and it was followed by the National Health Insurance in 1938, Seamen's Insurance in 1939, and Employees' Pension Insurance in 1921. However, these were seldom widely available in actual practice because of the characteristics of public assistance which limited them to the poor.

The concentration of catalase protein in anemic blood with enhanced population of reticulocytes and in non-anemic blood was determined immunologically by double diffusion test with anti-mome-liver catalase rabbit serum. The change in catalase protein concentration in anemic blood during incubation at 37°C for 24 hours was also studied. It was indicated that the diminished catalase activity in acatalasemic blood was due to the depletion of the protein and that catalase protein in acatalasemic reticulocytes decreased markedly by in vitro maturation. Furthermore, the possible presence of inactive catalase protein in acatalasemic blood was also suggested. Catalase protein concentration of acatalasemic anemic blood decreased by the incubation at 37°C for 24 hours in parallel with the decrease in reticulocyte count and catalase activity, and the decrease in catalase protein concentration of hemolysate by the same incubation parallel with the decrease in catalase activity. It is hypothesized that the unstable catalase protein with genetical change in structure easily decomposes during acatalasemic reticulocyte maturation is presented.

In order to examine how long diploid cells can be maintained in vitro cultivation without any chromosomal aberrations and to evaluate the mechanism of chromosomal heteroploid transformation, changes of chromosomes were studied in the course of serial in vitro transfers in four normal rat liver cell lines. As a result the diploid cells decreased in number gradually at early culture stage and disappeared completely in the periods between 350 and 500 days. The culture shifted to heteroploid as follow: diploidy---tpseudodiploidy---thypodiploidy---thypertriploidy or hypotetraploidy. This proces to heteroploid transformation is divided into five stages according to ploidy: 1- and II-stages show diploidy, III-stage, pseudodiploidy, IVstage, hypodiploidy and V-stage, hypertriploidy or hypotetraploidy. Chromosomal heteroploid transformation and neoploastic conversion occurred in IV-stage of the cultures. A possible mechanism involved in the process of chromosomal transformation was discussed.

Three inoperable patients with primary hepatoma could be placed on gluconeogenic diets (minimum carbohydrate-high fat diets) for one to three months. A transient inhibition or a marked retardation of the tumor growth was observed with these patients and their entire clinical courses were fairly good. These results confirmed our previous observation with a metastatic liver tumor patient.

By subcutaneous inoculation of N, N'-dimethylnitrosourea to adult male C3Hf/Bi mice once a week for 10 consecutive weeks the authors studied the correlation between immunological functions and histological changes in lymphatic tissues at the latent period of thymic lymphoma whose development is known to occur in 100 per cent. As a result, it was found that PFC of the spleen to sheep erythrocytes decreased to about one third the normal level by two weeks, and to one tenth by 8 weeks after initial inoculation of this compound. Hemolysin and hemagglutinin titers of the serum became less than 1 : 2 after 6 weeks and later. As for histological changes in the thymus, disappearance of lymphocytes became marked by 2 weeks, and there appeared tumor cells by 8 weeks. Also the peripheral lymphocytes as well as the total spleen cells decreased in number along with increase of the frequency of inoculation of N,N'-dimethylnitrosourea. These results seem to suggest that the immunosuppressive effect of carcinogen facilitates the development and proliferation of tumor cells possessing tumor specific antigenicity in the course of N, N'-dimethylnitrosourea- carcinogenesis.

As the first step to analyze the autoimmune disease of red cells the recognition mechanism of macrophage to red cells or erythrophagocytosis has been studied in vitro by using mouse peritoneal macrophage and homologous and cells and the following results were obtained: 1. In Hanks solution, the mouse macrophage hardly phagocytizes living red cells, both homologous and heterologous ones. But in the presence of mouse serum, the macrophage phagocytizes heterologous red cells selectively but does not phagocytize homologous ones. 2. The macrophage actively phagocytized homologous red cells prior to treatment with concanavalin A (Con A) at a concentration as low as 1.95 ltg/ml. 3. Red cell agglutination was clearly recognized in those treated with Con A at 62.5 lag/ml or more, but not at 1.95 ltg/ml. 4. The red cell agglutination by Con A was inhibited with D-glucose, D.mannose and a-methyl glucopyranoside at the concentration as low 1.5 mM, while the phagocytosis was suppressed only at a very high concentration of the sugars, 1, 000 mM. 5. Fragility test of the red cells treated with Con A showed a lower resistance of red cells to hypotonic solution than those treated with Con A at the concentration of 31.25 p.g/ml or more 6. Electron microscope observation revealed no membrane damage of red cells by treating with Con A at a concentration of 1.95 ,ag/ml, where erythrophagocytosis was observed. The membrane damage occur. red by treating with Con A at 31.25 ltg/ml or higher. 7. All the data indicate that the phagocytosis of homologous red cells by macrophage is induced by the adherence of a small amount of Con A, which induces no detectable changes of red cell surface and red cell membrane as revealed by agglutination test, fragility test, electron microscope observation and circular dichroism. On the basis of these observations a possible recognition error to homologous red cells by adsorbing a minute quantity of foreign substances on their surfaces has been discussed.

I) On an identified giant neurone of the right parietal ganglion in a snail's subesophageal ganglion-complex, the synaptic contribution to the production of the plateau formation of biopotential or grouped spike discharges of the soma has been studied in the presence of a convulsant. 2) The orthodromic stimulation of a peripheral nerve (the intestinal nerve) can elicit the plateau formation of biopotential, instead of normal spike discharges, in the identified neurone treated with a convulsant. 3) With the application of a convulsant, for example bemegride which was in a concentration less than that necessary to produce the plateau formation, an EPSP accompanied a spike with a constant delay. This EPSP is a product of a proprioceptive reflex arc consisting of two excitatory synapses with a certain subordinate neurone. 4) Later, in the presence of a convulsant, spontaneously conveyed multiple EPSP's were observed on the biopotential of the identified neurone. These multiple EPSP's produced grouped spike discharges or the plateau formation of biopotential of the neurone. 5) The multiple EPSP's may be produced by the grouped spike discharges of the subordinate neurone, the membrane property of which would be changed by a convulsant. It is presumed that the grouped spike discharges or the plateau formation of biopotential often occurs synchron. ously in many neighboring neurones by means of synaptic triggering in the presence of a convulsant.

A method of intracranial transplantation of the tumor induced by adenovirus type 12 in syrian hamster has been described. The incidence of intracranial tumor development was 86 (90.5 %) out of 95 animals and the average survival time and tumor size at death were 15.1 days and 4.1 mm in diameter respectively. The consistency of the days of death after intracranial transplantation of the tumor was remarkable. The transplanted tumors developed preferentially at the site of implantation and tumor cell seeding and tumor growing took place rarely along the ventricular system. Glial or lymphoid cell response to the tumor was not observed at any stage after transplantation in surrounding cerebral tissues of the animals. Histomorphologically, no elementary differences were observed between intracranially transplanted tumors and serially transplanted subcutaneous tumors. These facts permit the system to be applied to an experimental brain tumor model as large-scale testing.

This paper describes various aortic parabiotic procedures and discusses various problems concerning these procedures. The most satisfactory results, nearly 100 % of survival rate, can be achieved using longer sections of aortae of 2 to 5.month old rats. In these rats blood circulation between the parabionts has completely been established.

A subcutaneous tumor of a patient with epidermodysplasia verruciformis was studied by the light microscopy, the electronmicroscopy and the immunofluorescent test. The tumor cells were histologically pleomorphic and electronmicroscopically contained varying amounts of cytoplasmic filaments without Z-band formation. The antimyosin serum stained the tumor cells, showing their myogenic origin. No virus or virus-like particles were observed in the tumor. Tumor antigens stainable by the patient's serum were not detected. Hamsters inoculated with the tumor extract at birth developed no noticeable diseases.

Newborn mice of C3Hf/Bi (Zb) strain were divided into three groups and injected, intracranially with adenovirus type 12 alone, subcutaneously with 20 mgjkg of N, N'-dimethylnitrosourea following intracranial inoculation of adenovirus type 12, and subcutaneously with 20 mgjkg of N, N'-dimethylnitrosourea alone at 10 days of age, respectively. With adenovirus type 12 alone, intracranial tumors were induced in 12 out of the 25 effective animals. With N, N'-dimethylnitrosourea following adenovirus type 12, intracranial tumors were produced in 19 out of the 21 effective animals and these tumors were virus-induced ones. With N, N'-dimethylnitrosourea alone, no intracranial tumors were induced. In control mice, administered subcutaneously with 20 mgjkg of N, N'.dimethylnitrosourea within 24 hr after birth, necrosis of the external granular cells and hypoplasia of the granular layer of the cerebellum was observed.

As a step in the elucidation of human cancer immunity we studied antitumor activity of lymphoid cells by conducting a series of cultures using the primary culture of cells from spontaneous mammary cancers from C3H and RIll mice mixed with autochthonous lymphoid cells, and obtained the following results. 1) With 24 mammary tumors obtained from 24 mammary cancer. bearing mice, we prepared 22 suspensions containing sufficient numbers of free tumor cells, and attempted primary culture with them. As a result we were able to attain satisfactory primary culture cells in 18 trials. 2) With each group of the 18 primary culture tumor cells we conducted mixed cultures with autochthonous lymphoid cells (mainly spleen cells) in proportion of 1 : 40, for 48 hours, and counted viable tumor cells after the culture. As a result it was found that in 11 trials the lymphoid cells showed antitumor activity. In the remaining 7 groups of lymphoid cells there could be observed no antitumor activity, but some of them showed tendency to slightly accelerate the growth of tumor cells. 3) On looking at the correlation between the antitumor activity of lymphoid cells and the ratio of tumor weight/body weight, it was revealed that the antitumor activity is greatest when the tumor is around 10% the body weight, and as the tumor grows larger, such antitumor activity disappears. From these results, it may be concluded that even in spontaneous mammary cancer of mouse, autochthonous lymphoid cells exhibit anti. tumor activity on indigenous tumor, and this seems to indicate that cell. mediated immunity has been established.