Okayama University Medical School

Summing up the above problems we may group them as follows: 1) Szirmai's angio-myograph and myotonometer furnish us with means to evaluate the author's successful method or medical treatment in thrombophlebitic, postthromboembolic (ulcer, etc···) states on the basis of the blood circulation through the muscles, clearly registered by the angio-myograph. 2) Szirmai's medical preparation "HAH" serves as a quick and effective cure for thrombophlebitis. Results are very often reached within a few days. The patient's health is restored so as to make him able to work. 3) The above preparation assures full success in the cure of thrombotic esp. thromboembolic states of the lower limbs-cases of ulcus cruris includedwhich up to now could not be favourably influenced by any other method of treatment. Description of varieties of above problems and other types of cases of peripheral circulation (Endangitis, etc.) and their relationships with the subject will be given in additional papers. The author reports on registering and controlling thrombophlebitis, postthromboembolic states, including ulcus cruris, origin~ting either in above morbid conditions or in independent causes by means of the angio-myograph and myotonometer devised by the author. The reader is made familiar with the author's (Szirmai's) preparation "HAH" (Heacrin) and with the results achieved by applying it for the cure of acute thrombophlebitis and thrombotic states. Results are often showing up remarkably soon (2 to 6 days).

The activities of five hydrolytic enzymes, alkaline and acid phosphataSe, beta-esterase, leucine aminopeptidase and beta-glucuronidase, of human gastric carcinomas from 180 patients were investigated histochemically. Alkaline phosphatase activity was almost negative in the carcinoma but was weakly positive in this tumor at times (about 10 to 20 per cent). Acid phosphatase activity which displayed a slightly increasing tendency of the reaction in poorly differentiated tumor was variegated and mainly from feeble to moderate in activity. Beta-esterase reaction was in varying degrees with each case, but more malignant the carcinomas, the weaker was the activity. Leucine aminopeptidase was positive in about 30 to 60 per cent of the specimens observed but the reaction was founded to be localized often in some areas and generally similar to alkaline phosphatase reaction. The activities of leucine aminopeptidase, alkaline phosphatase and beta-esterase were positive at a higher rate in mucinous carcinomas than in non-mucin producing one. Beta-glucuronidase activity was slight or moderate in general but rather strong in the early stage of carcinomas.

In the present communication the recent works done by the Rheumatism Research Group of Department of Orthopedic Surgery, Okayama University, are described. The principal findings may briefly be summarized as follows. 1. Pathohistological pictures of the synovial membrane are classified into six types. Among them, Fibrinoid type and Follicular-Fibrosis type are the representative ones of chronic rheumatoid arthritis. 2. For the evaluation of the systemic as well as the local activities in rheumatoid arthritis and for judging the therapeutic effect, some indices have been established. 3. Injection of steroid hormones into the local joints fails to give satisfactory results in advanced, chronic rheumatoid arthritis. In such instances the flushing of the joint with physiological saline solution is effective. 4. In the case of chronic rheumatoid arthritis where the inflammation of hand and phalangeal joints is marked, RA-test gives rapid and more intense reaction, and most of such cases are of Follicular-Fibrosis type. 5. When lymph follicles appearing in the synovial membrane are stained when methyl green pyronine, the arrangement of lymphoid cells and plasma cells becomes distinctly clear. By micro-autoradiographic observations it can be seen that ³H-thymindine injected into the joint cavity is mostly ingested by the lymphoid cells in lymph follicles. 6. In the observation by the fluorescent antibody method multinuclear leucocytes found in the joint fluid and in the peripheral blood react with 19S and 7S-gamma-globulins. 7. When the serum and the joint fluid of the patient with rheumatoid arthritis are fractionated, they separate into three peaks at 19S, 7S, and 4S. Both S. S. C. A.-test and L. F. T. tests reveal the peak at 19S. The serum of chronic hepatitis positive to RA-test and the serum of rheumatoid arthritis are found to react immunologically the same to anti-β2 M globulin sheep serum. 8. When the reticulo-endothelial system of rat is blocked by 900,000 molecules of poly-vinyl-pyrroridon, the ability of antibody production is diminished. 9. Chemical synovectomy of injecting osmic acid is effective to FibrinoidCoating type. Its action mechanism lies in the complete cleaning of the surface of synovial membrane. 10. By radiating synovectomy with 193Au a fairly good result can be expected. 198Au is ingested by those cells in the surface layer of the synovial membrane and also by histiocytes in the synovial membrane. When 5 mc of 198Au are injected into the knee joint, a marked necrosis of the synovial membrane occurs. When 198Au is added to the ascites cells of rabbit during the tissue culture, in the concentration of over 14 μC degeneration of these cells can be recognized. 11. From the examination results of prognosis on those 25 cases with 41 rheumatoid knee joints after surgical synovectomy, it is considered that this method is indicated for Follicular-Fibrosis type. Ones with rheumatoid knee joint of Fibrinoid-Coating type gold sol treatment should be resorted to. In the cases of hand joints, surgical synovectemy is to be recommended at a relatively early stage.

The present report describes the findings on the infectivity of DNA partially purified from SV-40 which was propagated in the monkey kidney cells (BSC-1) in vitro and the importance of nucleic acids as oncogenic factors, particularly the induction of tumor by DNA in newborn hamsters. 593 newborn hamsters in total were used in the present experiments, and cannibalism among them posed as a serious problem. On 30 days postinoculation, very remarkable changes occurred in the liver, lung and subcutaneous areas. Cellular responses of the perivascular cells were predominant. and they were distributed in the interstitial tissues of the liver (liver cirrhosis in primates) and lung. Three hamsters of those subcutaneously inoculated with nucleic acids developed tumors and two tumors appeared in the subcutaneous tissues on 130 days postinoculation, which were identified to be the ones induced by intact SV-40 virus. Other tumors appeared in the liver, lung, intestinal ducts and abdominal surface at 126 days after subcutaneous injection. The cytological observations revealed multiple hemangiosarcoma combined with proliferation of the perivascular cells. On the other hand, cellular responses to nucleic acids were more marked by inoculation of the cell-free filtrate of BSC-1 infected by DNA than of DNA, and essential histologic findings were similar to the respo.nse to infectious DNA. Thirty-nine hamsters (30 per cent) developed tumor within about 200 days postinoculation of the filtrates. Sarcomas were common and they were confined to the subcutaneous tissues in 35 hamsters and to the peritoneum in two others by subcutaneous inoculation of the filtrates. The intestinal gland-cell carcinomas, however, could be induced at 37 and 59 days postinoculation in two hamsters of one litter (7 newborn hamsters) and in the other three newborn hamsters subcutaneous sarcomas were induced by inoculation of the same agent. These results suggest that the observation on the oncogenic capacity of nucleic acids would give us a clue to resolve the course of cancer from the view point of the infectious nucleic acid.

The distribution and activities of five hydrolytic and eight oxidative enzymes were histochemically studied in 60 different tumors of the human intestines. Benign polyp showed similar activities of most enzymes as those in normal crypt cells of large intestine with exception of higher activity of succinic dehydrogenase in benign polyp than in crypt cells. Malignant polyp had higher activities of most oxidative enzymes. Reticulo-sarcoma had weak activities of all enzymes. Carcinoid had strong activities of glucose-6-phosphate dehydrogenase and isocitric dehydrogenase while very weak of succinic dehydrogenase. Carcinoma showed varying degrees of the activity of all enzymes. Alkaline phosphatase and aminopeptidase were almost negative in all cells but in the stromal elements their weak activities were sporadically observed. Most enzymes were decreased in the central area of the carcinoma cell nestle, while in the infiltrating area or in the margin of cell nestle they were not decreased and sometimes increased.

1. The ratios of free 5α-cholestan-3β-ol and cholesterol and esterified 5α-cholestan-3β-ol were higher in pylorus than in cardia. 2. Esterified cholesterol level was higher in cardia than in pylorus. 3. Among the stomach cancer tissues examined free cholesterol level was higher than in the non-cancerous. 4. Esterified 5α-cholestan-3β-o1 and cholesterol levels were lower in the cancerous tissues than in the non cancerous.

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.

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.

Adenovirus type 12 is of human origin and it shows carcinogenic activity in experimental animals. With negatively stained particles of this virus electron microscopic observations were carried out. As the result it was demonstrated that its capsid, like other adenoviruses, is an icosahedron and each capsomere is of a hexagonal shape with a hollow in its center, each of which is surrounded by 6 adjacent capsomeres and is composed of numerous small subunit-like-particles.

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.

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.

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 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 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.

From the histochemical study of the intercostal muscles of cat, the following results were obtained. 1. Three different types of muscle fibers have been clearly distinguished in intercostal muscles by histochemical demonstration of succinic dehydrogenase; namely, the white fibers are stained faintly, while the red fibers deep blue and the intermediate fibers purple or bluish purple. 2. The difference in these stains is due to the degree of the enzyme activity, i. e., the areas of high SDH activity are stained deep blue while those of relatively low SDH activity are stained purple. 3. At oil immersion magnification, the differences among the three types of fibers are clearly distinguishable by the amount, size, distribution pattern and subsarcolemmal precipitation of Nitro-BT formazan particles. 4. Concerning the spatial distribution of these three types of fibers in each intercostal muscle, the muscles in the cranial and caudal parts of thorax (I-IV, VIII-XII) show a higher proportion of red fibers, while those in the middle thorax show a higher proportion of white fibers. 5. The vertebral portion of the first internal intercostal muscle is composed of only two types of fibers, red and intermediate ones, and their diameters are almost the same in size as in soleus muscle. In the middle intercostal muscle (V-VII), an intimate relationship can clearly be observed between the size and the enzyme activity of muscle fibers as in the gastrocnemius muscle. 6. In comparison with the anatomy of thorax and the distribution of muscle fibers, it may be presumed that there is a close relationship between the distribution and the scope of thorax movements, however, no definite relation between the distribution pattern and respiratory participation of muscle fibers. 7. Hence, it appears that the intercostal muscles in the cranial and caudal parts of thorax perform original respiratory movements, while the muscles in the middle thorax mainly perform voluntary respiratory movements, perhaps display their function during forced breathing. The intermediate fibers may usually have some tonus and carry out the role of resisting ribs from falling inside by negative pressure of the thoracic cavity.

From the histochemical study on the vascular distribution of limb muscles of cats and rats, the following results were obtained. 1. The red, white and intermediate fibers of the soleus and gastrocnemius individually present dissimilar patterns of the vascular distribution. In the transverse section of the muscle, most of the capillaries are found preferentially assembled around the small red fiber. Usually the red fibers are supplied with 4-6 capillaries, while the intermediate fibers 3-4 capillaries and the white 1-2 capillaries. 2. In the longitudinal section, the capillaries run parallel with the muscle fiber. Occasionally, these capillaries branching out for anastomoses cross the fiber almost at right angle. These anastomoses are more frequently observed on the red fibers. 3. The soleus muscle is better supplied with capillaries than is the gastrocnemius, because this muscle is composed only of two types of capillary rich fibers, namely, the red and intermediate fibers. 4. The mean capillary-fiber ratio of the cat gastrocnemius proves to be 0.6 and that of the soleus 1.4. 5. From these observations, it is considered that there is an intimate relationship between the capillary distribution pattern and the type of individual muscle fibers. 6. The red fibers being with a high oxidative enzyme activity and rich in mitochondria may require an abundant supply of oxygen and have a greater number of capillaries. In contrast to this, the white fibers with a lower enzyme activity and a fewer mitochondria need not to consume so much oxygen and are supplied with a fewer capillaries, because they participate only in phasic contraction. The intermediate fibers, with a moderate enzyme activity, may be considered to have a moderate number of capillaries.

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.

As is generally known, the energy transducing reaction in mitochondria is of highly complicated one. Free energy produced by transferring electrons from substrate to oxygen, where many dehydrogenases and respiratory chain of mitochondria are concerned, is transduced to ATP formation or utilized for the ion accmulation reaction, synthesis of various substances, reversal electron transport and the mechanochemical changes of mitochondria. The mechanism of these energy trasducing reactions which is supposed to be closely related with each other, has not yet been clarified. The authors tried to solve these biological energy transducing mechnism by applying physical circuit theory in electronics and elucidate that the energy transduction occurring in mitochondria can be explained theoretically. And some unknown but possible reaction have been postulated from such a physical consideration.