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In cell cultures of Detroit 6, KB, and HeLa cells, treatment with certain amounts of 5-Fluorouracil resulted in the appearance of a strikingly distinct halo or chromophobic area, entirely encircling the compacted or contracted nucleoli, before the ultimate disintegration of the cells.

For the purpose to clarify the control mechanism of erythroid cell differentiation, the author observed morphologic changes in bone-marrow cells and circulating red cells in phenylhydrazine anemia of rabbits by introducing a mass of red cells into vein at one time and reached the following conclusions. 1. After red cell transfusion in a mass to animal showing a marked hematopoietic activity, anisocytosis or macrocytosis becomes distinct with the appearance of big reticulocytes and red cells as large as four times the normal in volume. This suggests, judging from their volume, the accelerated denucleation of erythroblast as early as at the late basophilic stage. 2. Observations on bone marrow at this stage revealed the reduction in the number of erythroblasts of undifferentiated type with the increase of rather differentiated ones. In erythroid islet, undifferentiated cells are found surrounding a reticulum cell located in the center, while well differentiated ones in the outskirt area are situated near the sinusoid. Such a cell arrangement suggests that the erythroid cell requires a high oxygen tension for its differentiation. 3. From these observations and other results obtained from the studies on reticulocyte maturation and RNA synthesis of erythroblast, the author stresses that erythroid cells can differentiate as long as it is provided with a certain level of oxygen, even though it may develop m-RNA for differentiation. In other words, there should be two steps in the differentiation of erythroblast, the first is m-RNA synthesis induced by the information and the second is the somatic protein synthesis with oxygen supply. This seems to be directly connected to the control mechanism of hematopoiesis by oxygen.

For the purpose to reveal the role of R.E.S. for hemopoiesis and antibody formation, the R.E.S. of rabbits were severely blocked by the repeated intravenous injection of a vast amount of India ink, reaching 200 to 250 cc in total and the development of anemia and antibody formation by challenging egg albumin were observed while referring to the histologic changes in bone marrow, spleen and lymph nodes. The results were as follows: 1. The repeated intravenous injection of a vast amount of carbon particles induced a severe anemia. The anemia was always normo- or hyperchromic, showing not any disturbance in iron metabolism or hemoglobin formation. The data suggested that anemia is due to the arrest of reproduction of erythroblast or differentiation of the stem cells to erythroblasts, but not due to inhibition of the iron metabolism. 2. R.E.S. had no relation to the proliferation or the differentiation of granulocytes. 3. The functions of R.E.S. related to erythropoiesis and lymphopoiesis are affected by blocking independently of its phagocytic potency. In spite of a severe anemia, the phagocytic potency of R.E.S. could never be lowered and liver and spleen grew much larger in size and weight, showing that the phagocytic ability of R.E.S. is extremely resistant against such a blocking. 4. The serum antibody titer proved to be at the normal level, and no change of the antibody production in spite of heavy blocking of R.E.S. with India ink.

Two cases (Case I, 24-year old male, and Case II, 41-year old male) of liver cirrhosis after viral hepatitis have been described with a special emphasis on the distortion of the hepatic lobular architecture induced by hepatic hemodYnamic changes. Careful and precise clinical and laboratory examinations as well as peritoneoscopic examination with liver biopsy, particularly with vascular stereograms of liver biopsy tissue, have been successively carried outfrom stage of normal lobular architecture to early stage of cirrhosis. As the result, it has been found that in the course of these examinations clinical and laboratory features of the patients have remained almost unchanged in spite of gradual aggravation of morphological pictures. It is especially noteworthy that on vascular stereograms of liver biopsy tissue the parenchymal cells under the scarred portal tracts have suffered atrophic changes. Thus, three individual portal tracts of Case I have been gathered in a single connective tissue located on the distributing area of a scarred portal tract, whereas a central vein of Case II has moved from center to side of the scarred portal tract. In the late stage, these two cases ultimately turned to liver cirrhosis.

For the purpose to clarify whether or not the cells of regional lymph nodes and spleen of the tumor bearing individual develop the antitumor activity the author observed the proliferation of JTC-11 cells in vitro by mixing with the lymph-node and spleen cells from the mice bearing Ehrlich ascites tumor in solid form. After 24- to 48 -hour incubation the antitumor activity was estimated from the number of proliferated JTC-11 cells. As the result, it has been found that one week after implantation of tumor the regional lymph-node cells acquire the inhibitory activity against the proliferation of JTC-11 cells. The spleeen cells also show a marked inhibitory effect on the turner cell proliferation but two weeks after implantation these inhibitory activities of the cells both from lymph node and spleen are largely retarded three to four weeks when the host is emanciated by the growing tumors. Discussions are made on the inhibitory mechanisms from the viewpoint of immune reaction and on the transplantability of tumor cells without any rejection.

Almost all the umbilical lymphocytes showed more extensive blast cell formation than that of their mother's lymphocytes with PHA. Pathological conditions of mother in pregnancy and labor such as anemia, gestational toxicosis, difficult labor and asphyxia of babies, inhibited the normal response of both maternal and umbilical lymphocytes to PHA.

The mechanism of induction of anisocytosis was studied with experimental anemia of rabbits induced by blood depletion and phenylhydrazine chloride injection. The Price-Jones' curves of erythrocytes from anemic animals showed a large variety of red cell size, indicating the appearance of abnormally large sized cells. RNA contents of some reticulocytes in anemia were comparable to those of polychromatic and late basophilic erythroblasts. The result proved that in severe anemia a large number of erythroblasts are denucleated at earlier maturation stages, in most cases at polychromatic, and some at late basophilic and some at orthochromatic stages, resulting in anisocytosis.

Chemical and biological characteristics of the unsaturated fatty acids from the liver of irradiated and non-irradiated animals and some unsaturated fatty acids in sale have been described. The unsaturated fatty acid fractions from the rabbit liver taken after irradiating animal with x-ray show hardly any difference from those of non-irradiated animal in each component. But the former were distinguished from the latter in the increased rate of velocity of autoxidation. Similar characteristics were observed on the unsaturated fatty acids irradiated in vitro. They developed less labile free radicals with the shift of the double bonds to the carboxylic group and the conjugated double bonds, dienoic and trienoic acids. Biologically, the fatty acids from the irradiated animal suppressed the growth of bacteria requiring unsatturated fatty acid. And they are slightly stronger in the activity of uncoupling effect for the oxidative phosphorylation and the swelling of mitochondria comparing to those of general fatty acids, oleic and linoleic acids. They showed a strong lytic activity on the cell membrane as in the case of general fatty acids, linoleic, oleic, and some long chain unsaturated fatty acids. Tumor cells surviving through the treatment with unsaturated fatty acids changed the cell characteristics temporarily, with a slow-down of the ascites development and the cell growth.

In the experiments conducted with cells from the spleen and regional lymphoid cells derived from the axillary and regional lymph nodes of the mice receiving subcutaneous transplantation of Ehrlich tumor cells, it has been demonstrated that these lymphoid cells possess an ability to inhibit the proliferation of the subtrain of Ehrlich tumor cells (JTC-11), and that there is a parallel relationship between the rate of appearance of immunologicallly competent cells among these sensitized lymphoid cells and the inhibitory effect of these cells on JTC-11. In the experiment to reinforce the potency of sensitized cells conducted by means of the diffusion chamber technique, it has been proven that, while it is not an absolute prerogative, it is necessary to have the sensitized lymphoid cells come in contact with the target cells is order to wield the power of the former to the fullest extent. In the tissue cultures of lymphoid cells, it has been shown that sensitized lymphoid cells has a shorter life span than non-sensitized ones.

Macromolecular PVP was introduced intravenously into rabbits for a long period of 3 months, 10 g of PVP in total, and the observations were carried out to see disturbances in hematopoiesis, lymphopoiesis, and immune reaction with special reference to the histologic changes of marrow, liver, spleen and lymph nodes. The results were as follows: 1. A mild anemia was induced by the PVP injection. RE cells of liver and bone marrow were swollen moderately but otherwise no significant histologic changes were induced in bone marrow and liver. 2. A severe lymphocytopenia resulted: the RE cells of lymph follicles were blocked by PVP, the follicles collapsed into a homogeneous mass with fibrosis and minimized lymphopoietic tissues. These results suggest that RE cells of the so-called germinal centers are important for the reproduction or the differentiation of lymphocytes. 3. Intravenous injection of egg albumin caused the serum antibody formation with a marked proliferation of plasma cells around small vessels in lymph nodes and spleen as in the case of control animal. The data indicate that plasma cells are solely responsible for the serum-antibody formation and plasma cell may differentiate from adventitial cells of small vessels but not from lymphocyte or reticulum cell.

There is as yet no plausible and convincing explanation for the etiology of rheumatoid arthritis. The authors investigated anti-synovial tissue antibody in the body fluid of rheumatoid arthritis by means of indirect immunofluorescent technic using non-affected synovial tissues as antigen. As the result the anti-synovial tissue antibody was detected in 7 cases of the 15 synovial fluid samples of rheumatoid arthritis and in two out of the six serum samples. The site of the localization of this antibody was demonstrated to be in the synovial membrane, especially in synovial cells and in the small blood vessel walls situated immediately adjacent to the synovial surface, but it was found in no connective tissues other than synovial membrane. It seems that this anti-synovial tissue antibody should be considered as an independent factor from rheumatoid factor, and that rather than the rheumatoid factor it is more actively associated with the localization and progression of chronic inflammation within the rheumatoid arthritis joint.

Human brain turnors removed from 126 patients were histochemically examined and following results were obtained. 1. In general, alkaline phosphatase activity is decreased in poorly differentiated gliomas, but is not related to the tumor cell infiltration. 2. All the cases of alkaline phosphatase negative gliomas have poor reconvalescent course and most of the positive cases show good reconvalescence. 3. Alkaline phosphatase, leucine aminopeptidase and acid phosphatase activities are remarkable in fibroblastic meningioma, moderate or feeble in meningocytic meningioma, and negative in malignant meningioma. 4. The activities of alkaline phosphatase, β-esterase, leucine aminopeptidase and acid phosphatase are decreased in most of meningocytic meningiomas when the duration of symptoms and signs is short. 5. Succinic dehydrogenase, malic dehydrogenase, isocitric dehydrogenase and β-glucuronidase are strongly reactive in malignant meningioma; from strong to moderate in meningocytic meningioma and from moderate to feeble in fibroblastic meningioma. 6. There is a slight increasing tendency of the activities of succinic dehydrogenase, malic dehydrogenase, isocitric dehydrogenase in fibroblastic meningioma and p·glucuronidase for a short duration of symtoms and signs. 7. In the case of acoustic neurinomas the higher the alkaline phosphatase activity, the longer is the duration of symptoms and signs.

For the purpose to study in vivo changes of the mice bearing Ehrlich ascites tumor with special emphasis on the problems of cellular antibody and serum antibody, a series of experiments such as neutralization tests in vivo and in vitro study of the effect of lymph-node cells from the tumor bearing animals on target cells were carried out, and the findings thus obtained are briefly summarized as follows : 1. Regional lymph-node cells from the mouse transplanted with Ehrlich ascites tumor cells show a marked cytotoxic action on their cultured target cells, JTC-11, synergistically with serum from mouse bearing Ehrlich cancer. 2. The tumor cells inoculated with lymph-node cells from the tumor bearing animals showed a retardation in growth and finally regressed. 3. Spleen and lymph nodes of tumor bearing animals showed a marked increase in weight.

1. For the purpose to clarify the role of reticuloendothelial cells in the center of erythroblastic islet, the medullary and extramedullary erythropoietic foci in convalescence of phenylhydrazine arternia were observed by light and electron microscopies, and the mode of development of anemia in rabbit having blocked RES. 2. Light microscopic observations revealed the stimulated formation of erythroblastic islet in phenylhydrazine anemia. Electron microscopic observations revealed the rhopheocytotic vesicles in the membrane of erythroblast, some of which contained ferritin particles. 3. Repeated India ink injections induced anemia with RES damage by being heavily laden with soot particles. Color index of these animals was not reduced to the state of hypochromic anemia. Anemia is due to the damage of erythroidcell reproduction, but not due to the disturbed hemoglobin metabolism by the inhibited iron uptake. 4. From the results obtained the author concludes that the role of reticuloendothelial cell in islet is not to transfer ferritin to erythroblast but the reproduction of erythroid cells.

Adenovirus 12-induced tumor has been so far considered to be an undifferentiated sarcoma, but in the present study it has been possible to obtain such electronmicroscopic findings that substantiate well the theory of the neuro-ectodermal supporting cell origin as suggested by the observation at optical level. In other words, a specific clinging picture of cellular membranes and the presence of desmosomes have been demonstrated. In addition, though only in rare instances, the presence of virus-like particles have been verified, and some comments have been made about the relation between tumor and the appearance of virus as well as about carcinogenic mother cell.

Adversive movement was first reported in 1870 by Fritsch and Hitzig when they said that when gyrus Sylvii was stimulated electrically the eyes moved to the opposite side of the stimulation. Vogt and Foerster made a detailed report on adversive movement, which is a rotary motion of the head, trunk and both eyes. But in their report, they did not make it clear what tracts the stimulation took. Mitsueda, who iS under Hayashi, defined the cortical area of the eye balls and the eye lids. He reported that they were of the extrapyramidal kind. Russel reported that when the cerebellum was stimulated the eye balls moved to the side of the stimulation, but did not say anything about adveraive movements which concerned the movement of the head and the trunk. Therefore, to ascertain the center of adversive movement and its tract the following experiments were performed. For stimulation electrical ones and chemical ones using metrazol (cardiazol) were used. As Ishizuka, who is under Hayashi, has proved excitement is only seen when cardiazol is injected among the nerve cells at a certain concentration, and it is not seen when injected among the nerve fibres.

March of spasm in epileptic convulsions was first observed by Tackson in 1863, when he said that in certain epileptic convulsions there is a phenomenon, where the convulsion starts from a certain muscle group and gradually spreads to other muscle groups. He called this, "march of spasm" and reported that it spreads according to the arrangement of motor representations in Rolando's area of the cerebral cortex. Since then, many important studies concerning the cerebral motor cortex were performed and reported. Recently, when Erickson had brought out a method in measuring electroencephalographic waves, Jackson's theory has been acknowledged. In Japan, Hayashi and his school has made an extensive study on epileptic convulsion. They used nicotine, cardiazol and others as chemical stimulations and decided the conduction tract of epileptic convulsion in dogs. The characteristic part of chemical stimulation is that, the nelve cells excite themselves when it is injected directly among them in certain concentrations and do not excite themselves when performed among nerve fibers. This was proved by Ishizuka. We used this method in dogs to see what was the mechanism of this phenomenon, "march" which is seen in epileptic convulsions and what tracts they took for conduction. And as its result, we found new facts that the presence of the motor cortex was needed for the march of spasm, and the conduction tract descending from the lenticular nucleus were quite different from Hayashi and his school had previously reported.

Mononuclear cells from rabbit joint fluid were studied after synovitis was induced by various means, including the intra-articular injection of bacterial endotoxin or of aggregated human gamma globulin in normal rabbits, or of HGG in rabbits previously sensitized to this material. The large majority of mononuclear cells in all groups were monocytoid rather lymphocytoid, and these cells were most readily labeled with tritiated thymidine on the first day after injection. On day 2 and 3, the numbers of labeled cells decreased, except for the animals previously sensitized with HGG, in which there was an upswing of labeling on day 3. This upswing was associated with a considerable increase in numbers of cells resembing synovial cells, and may possibly be a reflection of synovial proliferation. Macrophages loaded with engulfed polymorphonuclear cells were observed in all experimental groups, a finding which emphasizes the lack of specificity of this reaction.

In vitro cell transformation of human embryo cells could be induced by the DNA purified from SV 40. The result shows clearly that cell transforms a part of viral DNA into the genome. In addition, for the purpose of clarifying th~ biological differences between the normal and transformants the alteration of cell membraneous structures of transformants (hamster and mouse fibroblasts) were observed from 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 differ~nce 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 that cell membranes of transformants are altered in the molecular structure r~sponsible for the surface charge. In addition, there is no remarkable quantative differences of sialic acids on the cell surfaces of non-malignant and malignant transformants so that phagocytic activity might be correlated to the alteration of molecular composition of cell membrane itself rather than of cell surfaces, i. e, sialic acids.

The incorporation of tritiated thymidine into DNA molecules was studied by electr.:m microscopic autoradiography. To make autoradiogram in electron microscopic level, DNA was extracted from rat ascites hepatoma (AH 130) cells after in vitro incubation with tritiated thymidine. Extracted DNA samples were rotary shadowed with platinum palladium and covered with emulsion. Silver grains demonstrated on autoradiogram indicated tritiated thymidine to have incorporated into DNA molecules themselves. The incorporation was further confirmed by liquid scintillation counting of TeA soluble and insoluble fractions after DNase or RNase treatment of the DNA preparations.