Okayama University Medical School

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.

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.

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.

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.

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.

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.

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.

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.

We succeeded in the extraction of a substance from beef cornea and rabbit muscle, that markedly inhibits mitosis of sea urchin eggs. The substance extracted from beef cornea is non-dialysable and it can be separated into three fractions by DEAE-cellulose column. Although everyone of these fractions has an antimitotic action, that of fractions II and III is especially marked. These fractions are one of nucleoproteins that have adenine as base. The substance extracted from rabbit muscle is dialysable, and when it is fractionated through DEAE-cellulose column into three fractions, fraction I has no antimitotic effect but fractions II and III have it. Fraction II is one of nucleoproteins that have hypoxanthine as base. Carnin obtained from beef cornea or from rabbit muscle shows a typical protein wave, but after being treated with gas by passing oxygen through cornin solution the wave height is lowered. Carnin, however, is a very stable substance when kept dry in a desiccator.

According to the data obtained in this experiment by means of the geldiffusion technique, the specific antigen was not detected in MH134 ascitic tumor, comparing the anti-C3H liver sera with anti-MH 134 tumor sera, though a loss of organ specific antigen and weak antigenicity were found in MH134 tumor extract. In order to detect some qualitative alteration, supposedly a gain in antigenic components, the transplant rejection test was carried out. The result of this test indicates that the relative not absolute resistance could be induced to C3H mice by this prior sensitization with cell free active extract eluted from MH134 tumor tissue by Fluorocarbon treatment. During these experiments, it became clear that MH134 ascitic tumor cell has weak immunizing properties so that prolonged lapse of time and large dose of antigen are inevitably necessary. Moreover, through Fluorocarbon treatment of the tumor homogenate, the cellfree, serologically active antigen could be obtained, which will serve well for the induction of the isologous immunization.

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.

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.

1. For the purpose to clarify the relationship between the structural change and lipid composition of isolated rat liver mitochondria, lipid composition and swelling rate of mitochondria obtained from the rat of 3'-Me-DAB feeding and raised in cold room are measured, and the following results were obtained. 2. The mitochondria obtained from the liver of 3'-Me-DAB-fed rat and of rat raised in cold room show a low rate of swelling by addition of Na-oleate accompanied by the decrease in highly unsaturated fatty acids (C18:3 and C20:3or 4) and with the increase in saturated fatty acids (C16 and C18). 3. Activation energy for the mitochondrial swelling is about 16.2 Kcal in the mitochondria obtained from normal rat liver, but requires 19.7 Kcal in the mitochondria that show a low rate of swelling. The fatty acid composition, especially in glycerophosphatides which occupy about 80 per cent of total lipids, is a structural component of mitochondrial membrane, undergoes the change from former to latter in the following fashion: C16:0 21.73→32.10, C16:1 3.37→2.96, C18:0 25.0→29.75, C18:1 13.75→17.40, C18:2 23.90→16.0 and C20:3 or 4 12.23→1.79. 4. At the time of low rate swelling of mitochondria isolated from 3'-MeDAB- fed rat liver, there could be observed a marked increase of the acetone soluble lipid (simple lipids) in the total liver lipids and in the fatty acid distribution of the acetone-soluble lipids, oleic acid was markedly increased (0.838→3.81%/dry liver), despite the fact that in the acetone-insoluble fractions or in the mitochondria there are no marked changes in the oleic acid contents (1.84→2.56% or 0.212→0.246%/dry liver).

The swelling-shrinkage and oxidative phosphorylation of rat liver mitochondria affected by 3'-methy1-DAB feeding were observed in correlation with function by the method mentioned, and the following results were obtained: 1. By feeding 3'-methy1-DAB the swelling-shrinkage ability of rat liver mitochondria showed a remarkable alteration reducing in the amplitude. It reduced gradually during the days of feeding, reached the minimum value on 30th day and restored gradually thereafter (in Case 1). 2. ADP/O ratio also decreased by feeding the carcinogen reached the minimum point on 30th day and increased on 38th day showing the similar tendency in the decrease of the swelling-shrinkage amplitude (in Case 1). 3. The mitochondria from the hepatoma, which was induced by 3'-methy1DAB feeding, showed a lower amplitude in swelling-shrinkage with the dropped ADP/O ratio compared with those of mitochondria from liver tissue neighbouring the tumor. 4. The mechanism in the reduction of swelling-shrinkage ability has been discussed in the relation with fatty acid composition of mitochondria which is reported elsewhere. 5. From the above results it is deduced that lowered ability for swellingshrinkage with the reduced oxidative phosphorylation will be somehow related to the mechanism of cancer induction.

We experienced a case of eosinophilic granuloma in soft tissue, and demonstrated its patterns of hydrolytic and oxidative enzymes histochemically. Neutrophils were rich in acid phosphatase and glucose-6-phosphate dehydrogenase. Eosinophils had much acid phosphatase and less other hydrolytic and oxidative enzymes. Lymphocytes showed weak reaction in all enzymes. Lymph follicles and histiocytes or fibrocytes had moderately oxidative enzymes. Small blood vessels and collagen fibers were rich in alkaline phosphatase and had a moderate amount of oxidative enzymes and acid phosphatase.

Liver homogenates could synthetize S-(isopropylcarboxymethyl) glutathione (GSIV) from isovaleric acid and glutathione, and GSIV thus formed was cleaved into L-allo-isovalthine by kidney glutathionase preparation. Isovaleric acid-l-C14 incorporated into GSIV without prior cleavage by the in vitro system. The discrepancy of configuration between urinary and in vitro synthesized isovalthine was discussed.

1. The forms of irritation causing inflammation and pain are reviewed, with reference to the significance of histamine, serotonin and bradykinin and in particular to the interrelationship between inflammation and pain. 2. The various types of experimental pain are reviewed and mention is made of the human and animal analgesia test methods derived from them. 3. More detailed descriptions are given of the analgesia test methods used by us, namely: a) Silver nitrate gonarthritis-pain, rat, in which both strong and weak analgesics with an anti-inflammatory action are effective. b) Phenylquinone-induced abdominal pain, mouse, in which all the analgesics and anti inflammatory agents mentioned in this article are effective in a greater or lesser degree. c) Tail-flick and hot-plate tests, mouse, in which the strong analgesics, the weaker analgesics and the anti-inflammatory agents, with the exception of the salicylates, are effective. d) Dental-pain test, guinea pig, which can be used to demonstrate the activity of the various analgesics, including the salicylates and also colchicine, which is not active in any other test. e) Pressure-pain, mouse, in which only the strong analgesics (narcotics) are effective. 4. The action of a large number of analgesics, anti-inflammatory agents and related drugs in the various analgesia-tests and in acute experimental inflammation is presented in tabular form. 5. It is concluded that the use of several pain and inflammation tests is essential for screening both analgesics for special indications (severe, mild pain, pain due to inflammation, etc.) and universal pain-killing drugs.