Okayama University Medical School

As described in the foregoing, a certain degree of desirable effect can be recognized in every instance of the present trials. However, in order to apply these methods in clinics it is necessary to carry out further studies on the mechanism that operates in bringing about such an effect, but this paper is presented as a preliminary report.

A case of paroxysmal bundle branch block, two to one right bundle branch block followed by intermittent right bundle branch block, which is associated with chronic cor pulmonale secondary to active, far advanced pulmonary tuberculosis, is presented. The incidence and mechanism of the paroxysmal bundle branch block have been discussed.

Spontaneous C3Hf lymphatic leukemia maintained in an ascites form was transplanted into newborn Wistar rats less than 24 hours old. Single subcutaneous or intraperitoneal inoculation of the neoplastic cells resulted in progressive tumor growth fatal to the heterologous hosts. Limited serial passage through newborn rats was performed. The intraperitoneally heterografted leukemia grew as massive lymphosarcoma predominantly in the adipose and connective tissue compartments with invasion of the neighboring organs but without leukemic manifestations. The characteristic behavior and histopathologic features of the transplanted disease are presented in comparison with the results of similar experiments reported in the literature.

1. When chicken sarcoma virus is serially inoculated on the mouse brain, it loses its carcinogenecity, but when it is inoculated on young chicken, granuloma develops in the liver and lung. When this granuloma is transplanted on adult chicken, a transplantable fibrosarcoma is obtained. 2. According to literature, the originaltumor of the Brown-Pearce cancer is a basal cell cancer, but that imported to Japan in 1953 presented a histological picture of carcinosarcoma. The metastasized tumor of the eye presents a purely cancer tissue, but when this is inoculated on the testis, carcinosarcoma is reproduced. It is therefore considered that the mother cell of the sarcoma is of host origin. 3. MY sarcoma is not a sarcoma, but is a spindle cell cancer. It might be a sarcoma which transformed into a cancer during serial transplantation, but perhaps it was originally a cancer but had been erroneously diagnosed as sarcoma. 4. The tumors we obtained by means of the feeding tests of Yoshida tumor all developed at organs other than those of the digestive tract. They are chiefly reticulo-sarcoma, but others which develop are malignant granuloma in the liver and lung, malignant adenoma in the kidney, papilloma of pelvis, and ependymoma in the cerebral ventricle. Since the discovery of the Yoshida tumor in 1943, serial transplantation has been conducted for 19 years with this tumor not only in Japan but also in foreign countries, but there has been no report to this date that a transformed strain has developed by cell transplantation. It therefore must be considered that the carcinogenesis observed in our feeding tests is a carcinogenesis due to a mechanism completely unlike that of cell transplantation. It has been confirmed by electron microscopy that in the early stage of transplantation of this tumor into the abdominal cavity there was an additional tumor growth due to the anaplastic proliferation of serous cells. 5. During the serial transplantation of viral tumors and/or virus dependent tumors, the tumor sometimes undergoes a morphological change. Though the cause of this is not yet sufficiently elucidated, it is suspected that there is some relationship with virus in the wide sense.

A single subcutaneous injection of 20-methylcholanthrene into newborn AKR mice less than 24 hours old resulted in the acceleration of the development of lymphocytic leukemia, and induction of subcutaneous sarcomas and multiple-lung adenomas. Morphological descriptions of the respective tumors were given. It is suggested that the lungs of newborn mice of strain AKR may prove to be a sensitive organ to evaluate carcinogenicity of certain carcinogenic compounds.

It has been found that the clinical tissue culture method devised in our laboratory for bone marrow is satisfactorily applicable to the in vitro study of human and animal splenic tissues. The present experiments have shown that the growth pattern, migration mode or cellular population of the cultured splenic cells is fairly characteristic of each disease condition, and the technic appears to be a valuable diagnostic aid in the practice of hematology. Direct vision biopsy with the laparoscope is safe and unattended by untoward side effects, enabling us to obtain excellent biopsy material from the enlarged spleen in various blood dyscrasias.

1. Attempts have been made to confirm how the formazan formation is affected in the presence of oxygen gas when the cells are incubated with neotetrazolium salt and the subsrtates for the enzymes to be tested. 2. In the cases of succinoxidase formazan formation is minimized under pure O2tension, it increases with decrease in O2 tension, and reaches its maximum value under N2gas. 3. This relationship between the oxygen tension and the diformazan formmation can likewise be observed even after pretreatment of the system with KCN. 4. In measuring enzyme activity of the DPN-diaphorase system with L-glutamate and DPN as substrate and NT as hydrogen acceptor, the same relationships between the oxygen tension and the NT-reduction can be seen as in succinoxidase system. 5. In the determination of enzyme activity of the cytochrome-c-cytochrome oxidase system with p-phenylene-diamine as substrate and NT as hydrogen acceptor, likewise the diformazan formation is markedly affected by oxygen tension and increased with the reduced oxygen tension but under pure Ns gas the value is reduced. When the systen is pretreated with KCN, however, the diformazan formation reveals its maximum value under pure nitrogen gas, the values of which correspond to those values of endogenous reaction without substrate. 6. The above results show that the neotetrazolium salt can compete with O2 as hydrogen acceptor, and less values of formazan formation may be obtained under higher oxygen tension and the higher values under lower oxygen tension independently from the true activity of the enzyme.

The effects of chloroquine on the growth and morphology of fibroblasts in tissue culture, and in vivo granulomas were investigated. As the result, the drug was shown to have a potent action to inhibit fibroblast growth, which has led to a possibility of its clinical use to patients with malignant tumors.

1. Rat liver mitochondria are swollen by inorganic phosphate in the medium of slightly hypotonic sucrose solution containing respiratory substrate and the mitochondrial swelling is inhibited or turned to shrink by ADP, respiratory inhibitor, anaerobiosis and uncoupler of oxidative phosphorylation. This mitochondrial swelling is not inhibited by the inhibitor of phosphorylating respiration such as oligomycin and tributyltin chloride. 2. Rat liver mitochondria are swollen by ATP in the presence of antimycin A, inorganic phosphate and 0.1 mM of CaCl2 and such a swelling is inhibited by oligomycin. 3. Accumulation of a small amont of P³² in acid soluble Pi fraction of rat liver mitochondria proceeds even in the medium containing neither ATP nor Ca++ but is inhibited by respiratory inhibitor, ATP, ADP and uncoupler of oxidative phosphorylation. The accumulation of P³² in mitochondria, however, is not inhibited by oligomycin. 4. The accumulation of P³² is induced by ATP in the presence of antimycin A and Ca++(O.l mM) and such an accumulation of P³² is inhibited by oligomycin. 5. It is suggested that the Pi-induced swelling of mitochondria is correlated to the accumulation of inorganic phosphate and both of them are tightly coupled to the initial step in the process of oxidative phosphoryaltion.

Based on our original concept, a fibroblast-inhibiting agent, chloroquine, was used against various animal tumors. Among transplanted animal tumors, the drug was most effective on relatively connective tissue-rich Bashford and Brown-Pearce tumors, as reflected by prolongation of life span, inhibition of tumor growth, inhibition of lowering of liver catalase activity, improvement of iron metabolism, increase of tumor necrosis, inhibition of connective tissue formation, and decrease of acid mucopolysaccharide. On the other hand, it was of little advantage in Ehrlich, Yoshida and MH134 tumors which contain little connective tissue, except for a decrease of the amount of ascites and ascites tumor cells in the former two tumors. These results indicate that chloroquine suppress the growth of the tumors relatively rich in connective tissue. This effect of chloroquine appears to be due to the primary attack of the stromal connective tissue of tumors being followed by the degeneration of tumor cells, though its probable anti-tumor activity by the indirect effects through its anti-inflammatory and systemic humoral activities should be taken into consideration.

Cytochemical observation of the activities of diphosphopyridine nucleotide diaphorase (DPNH-D), triphosphopyridine nucleotide diaphorase (TPNH-D), succinic dehydrogenase (SDH) and a-glycerophosphate dehydrogenase (α-GDH) of Ehrlich ascites carcinoma cells were made and following results were obtained. The smeared cells showed moderate reactions and no marked difference in the intensity among the individual cells. The free floating cells were stained relatively faint but showed the differences in the staining intensity in individual cells. In the presence of benzalkonium, the reaction intensity proved to be intermediate between the smeared cells and free floating cells without benzalkonium and the differences in the staining intensity in individual cells were more marked. Observations revealed that the reaction intensity changes closely corelated with the stage of mitotic cycle of each cell. Namely, DPNH-D activity of the tumor cells, which generally hihger than that of leucocytes, increased remarkably in the end stage of interphase and decreases abruptly in mitotic stage reaching the lowest level in metaphase. After the metaphse the activity increased slightly and it is kept at almost the same level during the first half of interphase. This enzyme is localized mainly in the granules of the eytoplasm. The activity of TPNH-D showed the similar localizations as those of DPNH-D, though the reaction intensity is lower than that of DPNH-D. The activity of SDH of the tmnor cells is lower than that of leucocytes and its diformazan granules are localized in mitochondria. Its activity decreases in the mitotic stage the lowest level in metaphase and in the followed interphase it is kept in a almost constant low level. α-GDH activity of the tumor is lower than that of SDH but show the similar localizations as the latter.

With the purpose to elucidate the cause and difference of blood fluidity in sudden death and natural one, we have observed the fibrinolysis of the blood in medico-legal and pathological autopsies by means of Fibrin Plate Method, a routine method devised in our laboratory. As the result it has been found that in the blood serum of sudden death and in some of natural deaths from tumors, leukemias, etc., the decrease in fibrinolytic activity is equivalent to the amount of proactivator that combined with the SK-like substance liberated into blood. On the other hand, in the blood of most of natural deaths, and in that bled from vessels and stored in body cavities, no natural fibrinolysis is observable and the same fibrinolytic activity with SK as normal one is demonstrated. Thus it is concluded that the cause of blood fluidity in sudden death is due to the fibrinolysis.

As has been well established, reticulocytes (RC) synthesize the species specific protein, globin, actively for about 24 hours or more till the time of their complete maturation1,2,3. This will be possible only in the presence of messenger RNA (m-RNA)4,5. Since the splendid hypothesis of m-RNA proposed by JACOB and MONOD6 for explaining the mechanism of the transfer of genetic information from nucleus to cytoplasm, it has largely been accepted through the numerous observations that followed7,8,9,10. However, the m-RNA hypothesis, which has been deduced by observing the protein synthesis in E. Coli, includes the meaning of labile RNA which is incessantly decomposed and newly synthesized to compensate the rapid degradation. As m-RNA cannot be synthesized in RC which have no detectable DNA, it has been supposed that the m-RNA of RC should be considerably stablell,12,13. Even in the denucleated cells, however, the RNA synthesis might be possible because Borsook reported the positive RNA synthesis of RC14, and this result has recently been reconfirmed by BURNY15.

Since the classic work of Ranvier, it is well known that the mammalian striated muscle is composed of two types of muscle fibers, i. e., the red and white muscle fibers. In the previous paper1 it has been reported that the limb muscle fibers of mammals can be divided into three types from their activities of the histochemically demonstrable oxidative enzymes. Namely, the small red muscle fibers had a higher activity of oxidative enzymes, the large white muscle fibers a lower activity and the third type of muscle fibers being called "medium fiber" or "intermediate fiber" showed an intermediate activity between those of the red and white muscle fibers.

Some bile acids (dehydrocholic, cholic, chenodeoxycholic, ursodeoxycholic, and deoxycholic acids), and some hypocholesterolemic agents (22, 25 diazacholestanol, 20,25-diazacholesterol, triparanol, and SKF 525-A) are the inducers of isovalthinuria in guinea pig. Administration of methionine appears to increase the pool of sulfur compound which participates in the formation of isovalthine. Cholesterol appears to have no enhancing effect on the induction activity of isovalthinuria inducers. The mechanism of isovalthine formation and the role of sulfur amino acids in lowering blood cholesterol are discussed.

The effect of various fatty acids on the swelling-contraction and oxidative phosphorylation of mitochondria from rat liver and Ehrlich ascites tumor cell have been studied and the results are as follows: 1. The swelling of rat liver mitochondria is induced by fatty acid. The extent of this uncoupling action is in the descending order of myristate, laurate, parlmitate, stearate and behenate in saturated fatty acid and linoleate, linoleneate, richinoleate and oleate in the unsaturated fatty acid. This swelling action is stronger with unsaturated fatty acids than that of saturated ones and cis form is stronger than trans form. 2. The uncoupling oxidative phosphorylation of rat liver mitochondria is also observed with these fatty acids and the activities are proportional to the degree of the swelling action. 3. The degree of swelling of rat liver mitochondria is proportional to the concentration of oleate and is inhibited by anaerobiosis and respiratory inhibitor except amytal. 4. The mitochondria swollen by fatty acid can be recontracted reversibly by ATP, Mg++ and bovine serum albumin. 5. The swelling action of sodium oleate is the strongest on mitochondria from rat liver, followed by those from the liver of Ehrlich ascites tumor bearing mouse, Ehrlich ascites tumor cells and solid Ehrlich tumor cells. 6. Sodium oleate inhibits the incorporation of 32p into ATP, ADP, GTP and UDPG in mitochondria.

By far the majority of studies and speculations on metabolism in man and animals have been concerned with the fate of proteins, fats, and carbohydrates. Even when interest was awakened in enzymes and vitamins, the transformation of the organic substances was still studied. Gradually, however, various methods of the analysis and measuring instruments used have been so perfected that other problems can now be included in the field of biological and medical researches, namely, the significance of inorganic substances for the living organism.

Histochemical studies of several oxidative enzymes of the muscle spindles in mouse limbs were carried out. Observations elucidated the existence of the three types of intrafusal fibers which can be distinguished from each other by the difference in the activity of oxidative enzymes as in the muscle fibers, i. e., the first one is the "red intrafusal fibers" and has the most intense activity of oxidative enzyme, the second one the "white intrafusal fibers" with a minimal activity, and the third, "medium intrafusal fibers" being distinguished by their oderate activity of oxidative enzymes from the others.

For the purpose to reveal the mechanism of the biological indirect action of X-rays the author has conducted the analysis of the compound lipids of the organs of the rabbits irradiated with X-rays. Silicic acid column chromatography was applied for the fractionation of the compound lipids. Component of each fraction was analyzed by silicic acid-impregnated paper chromatography and infra-red spectrophotometory. The result proved that the compound lipids showed some qualitative and 10+-+ quantitative changes. The changes occurred mainly in glycerophosphatides showing the formation of high level lyso-phosphatidylcholine and lysophosphatidylethanolamine. The fatty acid compositions of the compound lipids were determined by means of gas-liquid chromatography. Further, polyunsaturated fatty acids were analyzed as fatty acid bromide. Major differences found in fatty acid patterns included: in the irradiated group the amount of C18-monoenoic, C18-dienoic, C18-trienoic, C20-tetraenoic and C22-acids were decreased and of C18- and C18-monoenoic acids were increased. It has been elucidated that lysophosphatides not only posesses a strong hemolytic power but also it has an action to induce swelling of the rat liver mitochondria. And it has been suggested that the lysophosphatides and ethanolamine will be responsible, at least partially, for the cell damage induced by X-rays.