Okayama University Medical School

From the data presented in this communication, it might be concluded that a cancer specific substance, which can be demonstrated in gel diffusion, is present in human cancer tissue, common to various epithelial cancers of different individuals, although it may vary in its concentration. Needless to say, this substance is quite different from the so-called interspecies antigen or organ specific antigen, as proved by the present experiments. Furthermore, this substance can be eluted well by the Fluorocarbon treatment and it displays physically and chemically unstable characteristics. This substance is likely to be included in the microsome fraction and soluble fraction which was determined by gel diffusion technique. However, the association of this substance with other specific antigenic substances of human cancer, concerned with "delayed type skin reaction", "cytopathogenic antiserum against cancer cell", and "complement fixing antibody in serum of patients with cancer", has not been elucidated in this study.

Both the cornea and muscle cornins have no effect at all on oxygen uptake of tissue, and likewise they catalase affect Qctivity not in any way. The corneacornin has an effect to reduce P/O ratio to about one half, but the muscle cornin does not show such an effect. Both comins decrease thc incorporation of P³² into nucleic acid fraction and DNA synthesis. In the ultracentrifugal analysis of nucleic acids during development of sea urchin eggs, cornins inhibit the polymerization of nucleic acids. In addition, both of these comins depress the incorporation of P³² into DNA and ribosome RNA of regenerating rat liver. Both comins inhibit the increase of -SH quantities before the cleavage of sea urchin eggs.

The normal mitotic dog kidney cell division and the multinucleated giant cell formation and degeneration of the dog kidney cells infected with measles virus were observed by the phase-cinematography. It took only five minutes for the mitotic cell division. The cell assumed a spherical shape before mitosis, and the two divided cells grew to the flat cells on the bottle wall. The giant cell formation was definitely the result of cell fusion. The cellular contents of the multinucleated giant cell were exposed after buddings, and the cell itself died.

The author studied the hematopoietic disturbances of rabbit induced by saponin injection and drew the following conclusions: 1) By saponin injection, the structure of bone marrow is disintegrated and hematopoietic cells are released into the circulating blood forming extramedullary hematopietic foci mainly in liver and spleen. The main attacking point of saponin should be RES. Recovery of hematopoietic foci is associated with the recovery of RES. The most marked extramedullary hematopoiesis is found three days after the injection. Thereafter, bone-marrow hematopoiesis proceeds to recovery stage, during which hematopoietic foci in liver and spleen are preserved, especially those in spleen persist fairly for a long time. 2) Daily injections of India ink kept up over a long period of time after the treatment with saponin, prevent the recovery of anemia and bone-marrow hematopoiesis. The lymph nodes, whose RES escaped from the severe damage by India ink, keep the hematopoietic foci for a long time. 3) As far as hematopoiesis is concerned, there seems to be no functional differentiation among RE cells, though they seem to have a special function according to the organs to which they belong, e. g. antibody formation in lymph apparatus, hematopoiesis in bone marrow and red cell destruction in spleen.

Using double staining method of succinic dehydrogenase and cholinesterase, the structural differences of motor endplate in the red, the white and the intermediate muscle fibers of the mouse limb muscles were observed. The endplate of the white fiber had a large size and complicated interlacing structure. The endplate of the red fiber had a small size, simple and compact structure. The endplate of the intermediate fiber had a medium size and moderately developed structure.

1. By using a monolayer of tissue culture cells, JTC-3, JTC-3', JTC-3" and HeLa cells, the assays of neotetrazolium (NT) reduction at the cell level have been conducted on the cells without detaching the cells from the culture glass vessel wall. 2. It has been found that the activity of endogenous NT reductase is extremely high in the living tissue culture cells. The endogenous NT reductase activity is found to be in the descending order of HeLa > JTC-3' > JTC-3 > JTC-3". The endogenous NT reductase activity increased in the medium having a low level of proteins and decreased in the absence of yeast extract. 3. It has been demonstrated that most of the endogenous NT reduction in the JTC-3 cell groups takes place at the step of flavoprotein in the NADHdiaphorase system, and a portion of it occurs being coupled with the ubiquinone step. In contrast, in HeLa cell it is p:>stulated that aside from the NADH-diaphorase system, succinoxidase system is involved in this reaction. 4. The coupling site of the succinate NT reductase system with the terminal electron transport system in the JTC-3 cell groups also differs from that in HeLa cells. Namely, in JTC-3 cell groups about 50% of the coupling occurs at the step later than antimycin A sensitive step and the remaining 50 % most likely at the ubiquinone step~ whereas in the HeLa cell most of the coupling takes place at the step later than antimycin A sensitive step. 5. Although there can be observed variations in the activity of the succinate NT reductase, hardly any difference was observed among the JTC-3 cell groups in the rate of the reduced NT amounts at several coupling sites, revealing that the change in the composition of the culture medium does not bring about any essential change in the coupling sites and their mutual activity rates of succinate NT reductase system. 6. Both endogenous NT reductase activity and succinate NT reductase activity have been accelerated markedly by potassium cyanide. The result indicates that in living cell the electrons are not transferred to NT at the level of cytochrome oxidase and O2.

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.

1. For the elucidation of the mechanism of isovalthinuria induction, some precursors of cholesterol biosynthesis and some other compounds were tested in guinea pigs to know whether they induced isovalthinuria or not. 2. Among the cholesterol precursors tested, isovaleric, β-methylcrotonic, and β-methylglutaconic acids were found to be strong inducers of isovalthinuria, but acetic acid, acetoacetic acid, β-hydroxy-β-methylglutaric acid, squalene, α-ketoisocaproic acid, and leucine were not. 3. Among the other compounds tested, palmitic acid and L-cysteine were found to be weak inducers of isovalthinuria. 4. Some antagonistic phenomena were found to occur. The inducing effect of isovaleric acid was inhibited strongly by dexamethasone or dehydrocholic acid, and weakly by insulin. The inducing effect of dexamethasone was inhibited by dehydrocholic acid or thyroxine. The inducing effect of dehydrocholic acid was inhibited by insulin or dexamethasone. 5. Although ethionine was not an inducer in itself, it acted as a substitute for methionine or cysteine in the case of the sulfur deficiency produced by prolonged feeding on bile acid. 6. Isovalthinuria was observed by the administration of bromural, but this effect may be a detoxication but not induction,

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.

Es wurde ein Triparanol-Präparat bei Arteriosklerose an 34 Patienten ambulant untersucht. Der Autor hat beim gröβten Teil seiner Falle eine wesentliche objektive und subjektive auch registrierdare Besserung beobaehtet. Der Blutcholesterin und der Blutdruek sind nicht nur bei den erfolgreiehen Fällen, sondern auch bei den wenigen Patienten, die Triparanol nicht vertragen konnten, gesunken, Oszillometrische, angiomyologisehe Werte und Hauftemperatur sind besser geworden. Schwinde1, Grübeln, Unsicherheit, stenokardische Beschwerden, schwermütige Gedanken, Benommenheit u. a. Symptome haden sich nur selten und nur in kleinem Maβe gezeigt.

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.