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As a step in the elucidation of immunity of human cancer from the standpoint of homotransplantation immunity, we conducted mixed cultures of regional lymph node cells from C3H mouse isotransplanted with methylcholanthrene-induced sarcoma (MC-tumor) together with the primary culture MC-tumor cells, and observed the behaviors of these lymph node cells to the MC-tumor cells and compared the effects of these lymph node cells with lhose of normal mouse lymph node cells by counting the growth number of tumor cells and also by cinematography. As a result, it has been demonstrated that the regional lymph node cells from the mouse isotransplanted with the MC-tumor (2mm3 in size) acquire a strong antitumor activity by 14 days after the transplantation, but such antitumor activity diminishes and disappears in the terminal stage of cancer. When the number of these lymphocytes is increased, there can be observed some dosage effect, but no complete inhibition of the tumor growth can be attained. The cinematographic observations of these regional lymph node cells in the mixed culture with tumor cells reveal that lymphocytes of small to intermediate size aggregate onto the tumor cells and inhibit the movement of the latter.

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. Isovaleric acid-1-C14, -4-C14, or C14-CaC03 with or without non-isotopic isovaleric acid was orally administered to rats and the incorporation of these isotopes into liver cholesterol, fatty acid, or urinary isovalthine was examined. 2. Isopropyl group of isovaleric acid was more efficiently utilized for cholesterol synthesis than carboxyl group, and also for cholesterol synthesis than for fatty acid. These results indicate that isovaleric acid is cleaved into two fragments before it is utilized for cholesterol synthesis. 3. Carbon dioxide was used for the synthesis of liver cholesterol and of liver fatty acid. Isovaleric acid seems to enhance the incorporation of carbon dioxide into cholesterol. 4. All the experimental rats received isotopic or non-isotopic isovaleric acid excreted isovalthine, but no radioactivity was found in it. Thus, isovaleric acid residue of urinary isovalthine molecule is not derived from isovaleric acid administered, and carbon dioxide is not the carbon source of urinary isovalthine. 5. Suspicious metabolism of isovaleric acid or of carbon dioxide was discussed. 6. Isotopic isovalthine which was synthesized from (± ) α-bromoisovaleric acid-4-C14 is administered to rat and it was found that the isotope did not incorporate into cholesterol or fatty acid of liver and of brain. About 15% of isotopic isovalthine was recovered in urine up to the next day after injection. The large part of isovalthine was missing.

1) Three different types of muscle fibers were clearly distinguished in the intercostal muscles and the diaphragm of human, cat and rat by histochemical demonstration of oxidative enzymes, phosphorylase and glycogen. 2) The intercostal muscles and diaphragm each presented dissimilar patterns of the muscle fibers. The diaphragm did not show any definite correlation between the diameter and the histochemical reaction of muscle fibers but its red fibers indicated a more intense uptake of Nitro-BT formazan deposits in the subsarcolemmal region. In this conection, the relationship between the motor innervation and histochemical evaluation of the diaphragm was described. 3) Phosphorylase and glycogen showed reciprocal reactions to the oxidative enzyme activity. They were generally high in large fibers but low in small fibers, and moderate in intermediate fibers.

With gastric carcinomas the activities of eight dehydrogenases; succmlC, lactic, malic, α glycerophosphate, glutamic, β-hydroxybutyric, glucose-6-phosphate and isocitric dehydrogenase were statistically estimated. Principal findings may be briefly summarized as follows. These enzymatic activities differed considerably even in the same classification of carcinomas and generally ranged from strong to weak in the following order: lactic, malic, glucose-6-phosphate, isocitric, succinic, α-glycerophosphate, glutamic and β-hydroxybutyric dehydrogenase. The activities of adenocarcinomas were stronger than those in simple ones, and these were not related appreciably to cell differentiation in adenocarcinomas except succinic, glutamic, glucose-6-phosphate and isocitric dehydrogenase. As for succinic dehydrogenase and NAD-linked dehydrogenases except for lactic dehydrogenase, the activities were strongest in intestinal metaplasia and early mucosal carcinomas, the next being in benign adenomatous polyps and weakest in the other carcinomas. As for NADP-linked dehydrogenases and lactic dehydronase, the activities were also strongest in intestinal metaplasia and early carcinomas, the second in the other carcinomas and the third in the benign polyps. Generally, these dehydrogenase activities were strongest in free carcinoma cells in blood and lymph vessels and in actively growing part of several carcinomas and weakest in the central area of tumors, especially almost negative in the central necrotic area.

Les auteurs out effectue après une irradiation totale de 1200r de rats blancs des deux sexes des examens hématologiques à la suite d'irradiations ainsi que des examens physiologiques et des contrôles. Ils n'ont observe de modification importante des facteurs coagulants qu'au troisieme jour; cette modification était maximum avant la mort, c'est-à-dire au stade terminal. Les temps de coagulation naturelle ont beaucoup diminué, de même que ceux de la thrombine et ceux de la thrombine avec le bleu de toluidine, c'est-à-dire que l'héparine libérée ( = antithrombine semblable à l'héparine) a diminue. Pour les facteurs V et VII et en particulier pour la prothrombine on a observe un fort accroissement de la concentration. Les auteurs pensent que ceci est explicable par le fait que la décomposition des tissus pendant l'irradiation entraine la libération de kinase et d'autres activateurs dans la circulation sanguine, ce qui provoque une anoxemie des tissus. D'autres expériences sont en cours en collaboration avec de nombreux spécialistes et instituts.

In the present experiment, it has been noted that clonizing epithelial-like cells of the intestine 407 were more susceptible to SV-40 virus than normal fibroblasts in primary human cell cultures. In the early stage of the infection the cell growth was enhanced by the inoculation of DNA virus but many cells died, showing lysis characterized by CPE, clumping of chromatin and formation of inclusion bodies. On the other hand, the cells surviving infection have given rise to virus-free long term cultures and cellular responses to the virus characterized by cell proliferation which is. classified in four phases. (Phase. I: infection and cell alteration. Phase. II: crisis. Phase. III: fibro-reticulum cell formation. Phase. IV: recovery and proliferation). The most remarkable morphological characteristic was fibroblastic cell alteration from epithelial cells at 5 weeks of virus inoculation. By this study an interesting generalization of human epithelial-like cells can be made about the differentiation of the transformed cells in relation to SV-40 virus and it has been shown that an established human cell line is still susceptible to the reverting action of the SV-40 virus.

Chromatography on Sephadex G-200 was performed with the soluble fraction of homogenated rabbit liver, which was extracted with 0.1 M phosphate buffer containing 0.1 M NaCl. and the influences of autolysis on the soluble fraction of liver were also examined. The soluble fraction of liver was different from serum in molecular weight, in electrophoretic character and in components with sedimentation coefficients. The soluble fraction of liver was stable under the influence of Mg and K ions, and rather unstable in the presence of Na ions. Serum was fractionated in three main peaks. The soluble fraction of liver was fractionated in a similar pattern as of serum, but the first peak contained nucleic acid and lipoprotein. The second contained albumin. 32p radioactivity peaks of the stored sample appeared with change in patterns by autolysis from the original, and were observed wide based and continuous figures in retarded peaks. The correlations with the first peak and retarded peaks were represented by the analysis of phosphorus compounds and electrophoresis. In lipid analysis, both diglyceride and monoglyceride gradually decreased, and phospholipid pattern was observed to increase in retarded peaks by autolysis. Lipoprotein or lipid-albumin complex was gradually converted to smaller molecular weight compounds, and appeared in retarded peaks.

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 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.

For the purpose to clarify the relationship between production of humoral antibodies and cellular reactions of the lymphoid system to allogeneic-transplanted cells in mice, a study on cross sensitization was carried out between inbred A(H-2a) and C3H(H-2k) strain mice. The median survival time of skin of C3H transplanted to A (C3H-to-A) was 14.1 ± 1. 4 days, and of A transplanted to C3H(A-to-C3H) was 11.8± 1. 6 days. Repeated A cell injections to C3H induced the formation of humoral antibodies, whereas the C3H cell injections into A did not. In A-to-C3H and C3H-to-A combinations, the immunization induced an increase in white blood cell number in circulating blood successively with the repetition of the antigen injection, and organ weights increased in thymus, spleen, and liver but not in kidney. Weight increases in the organs of A treated with C3H cell injection were less in extent, comparing to those of C3H treated with A cells. Histologic observations revealed that the cellular proliferation in the lymphoid system including plasmocytic responses were obviously predominant in the C3H treated with A cells comparing to those in the A treated with C3H cells. Hemocytoblasts also increased during the immunization in both cases showing no significant differences between the two series of experiments. These cellular reactions were observed not only in the draining lymph nodes but also in the generalized lymphoid tissues. The results of the present study suggest that the definitive factor for producing humoral antibodies is in the differences of the homologous antigenicity between the donor and the recipient but not in the degree of sensitization, and the Dk in H-2 loci is not so strong in antigenicity as to elicit sufficient plasmocytic responses for the formation of humoral antibodies in C3H strain mouse.

As a link in the series of studies on tumor-specific immunity, in vitro inhibitory effect of sensitized isologous lymph-node cells on the proliferation of C3H mammary cancer was studied. For this purpose tissue culture was conducted with regional lymph-node cells obtained from truly isologous C3H mouse inoculated with A strain cells derived from C3H mouse mammary cancer along with A cells, and the following results were obtained. In the case of tissue culture with those lymph-node cells obtained from the groups of mice 10 days after the inoculation of 5 X 106 A cells, the inhibitory effect on the proliferation of A cells was most marked, followed by that of those taken on day 14, 7, and 5 decreasing in the order mentioned. In the case with those regional lymph-node cells obtained from mice which did not have recurrence of tumor 1 week after extirpation of 2-week old tumor, the inhibitory effect on proliferation of A cells was marked, with the regional lymph-node cells obtained two weeks after transplantation of 1 × 108 A cells there could be observed no inhibitory effect at all. This suggests that at a certain stage after implantation of such regional lymph- node cells there develops a specific anti-tumor activity in the host.

In the experiments with cultured liver cells it is very important to know whether or not the cells in vitro have the same properties and functions as in vivo. The purposes of this work were to investigate the functions of the cultured liver cells and to identify functionally the liver cells cultured by our present method with the parenchymal liver cells. At first, the albumin production of the cultured liver cells, one of the well known functions of the liver cells, was examined by the immunological methods, especially, the fluorescent antibody technique and the complement fixation test. Culture methods which could display the functions of the liver cells as much as possible were explored simultaneously. The results were as follows: 1. Albumin production was detected in the strain RLN·10 liver cells established from the liver tissues of a Donryu rat with immunofluorescent method and complement fixation test. This confirms that the cultured liver cells maintain the function to produce albumin and these cells have originated from the parenchymal liver cells. 2. Hepatoma strains (AH 66-TC-l, AH 7974-TC-l) also showed the albumin production but the extent of its production was less than that of the strain RLN-10. 3. In the short-term cultured liver cells, the albumin production was testified only slight in one month and was exhibited in a small amount in three months. 4. Every culture method examined exhibited no appreciable difference in the albumin production in the cultured liver cells.

With the purpose to prevent the dissemination and consequent metastasis of cancer cells at the time of operation we gave 10 mg of Mitomycin C per day for four consecutive days prior to surgical operation of gastric cancer (total of 322 patients), and this so-called adjuvant chemotherapy proved to be effective on the cases with serosal involvement and infiltrating type of cancer, irrespective of histological types. It also gave five-year survival rate of 35 per cent. However, to lymph nodes already metastasized, the adjuvant chemotherapy proved to be not effective.

Large doses of adenovirus type 12 were injected intraperitoneally into adult hamsters, and development of tumors and other pathological findings were studied in comparison with those in hamsters injected when newborn. Doses of 38~47 TCID60 per gram body weight produced tumors in 3 of 12 hamsters injected at 37~57 days of age. A dose of 170 TCID60 per gram body weight produced tumors in one of 18 hamsters injected at 61~71 days of age, but in none of 18 hamsters injected at 147~174 days of age, while the same dose per gram body weight produced tumors in 24 of 26 hamsters injected when newborn. In hamsters injected at adult ages, the number of tumors per animal decreased and the latent period for tumor development became very long as compared with those in hamsters injected when newborn. Regardless of the age at the time of injection, acute inflammatory change was observed in the peritoneum which later developed into various degrees of peritoneal adhesion. Adenovirus type 3 also induced the peritoneal adhesion. Histology of tumors was studied and discussed.

DNA synthesis and cell renewal of mouse intestinal epithelium were studied with radioautography after injection of thymidine-H³ to know the difference of the mode of epithelial cell generation relating to the different frequency of cancer developement in several parts of small and large intestines. Succinic dehydrogensase activity was also observed by histochemical method. Renewal time of the intestinal epithelium of mouse is about three days throughout the intestine with somewhat longer time in rectum and anus, and relatively shorter one in ileum compared to the other parts of the intestine. Daily regenerating rate was low in large intestine, especially in rectum and anus. Strong activity of succinic dehydrogenase appeared in the bottom of crypt and seems to be correlated to the active cell division. Epithelial cells in large intestine move very slowly upward and few of them seem to move to the opposite side or stay long time at one place. Intermitotic time is about 27 hours in small intestine and about 40 hours in large intestine. These suggest some relations between the mode of the epthelial cell renewal and cancer development. Because in human the frequency of cancer development is very high in large intestine, rectum and anus, and the epithelial renewal of these areas is supposed to be delayed similarly as in mice.

Oncogenesis of human adenovirus type 12 in the brain of rats was examined. Newborn rats of Sprague-Dawley and Donryu　strains were injected intracranially with human adenovirus type 12.　The incidence of intracranial tumors was 91% (30/33) in SpragueDawley　and 56% (14/25) in Donryu rats. Except for one tumor nodule　located in the parietal cortex of a Sprague.Dawley rat, all tumors　developed in the paraventricular areas or in the meninges. Tumors were quite similar histologically to those induced in hamsters and　mice resembling the undifferentiated human brain tumors such as medulloblastoma,　ependymoblastoma and embryonic gliomas. From　the histological features and primary sites of tumor development, it is　suggested that the tumors in the brain of rats induced by adenovirus type 12 originate from the embryonic cells in the　paraventricular area　and also from the undifferentiated supporting cells of the peripheral　nerves in the　leptomeninges.

A factor, cornin, inhibiting the growth of L cells cultured in monolayer was extracted from bovine liver with boiling water and was partially purified by gel filtration with Sephadex G-200. The factor was (1) precipitable with ethanol at the concentration between 70% and 90%, (2) impermeable through dializing memo brane, (3) eluted as the last peak at the gel filtration and (4) containing protein and RNA but no DNA.

To analyze the appearance of three forms of hepatitis B antigen-associated particles (HB Ag particles) and antigen-antibody (Ag-Ab) complexes in the sera of patients with various liver diseases, electron microscopic observations with the combinations of a variety of immunological assays were made at first on the HB Ag and Ab mixed in vitro in various ratios, and then on the samples from the sera of each patient. The number of patients observed were 64 in total, which consisted of various types of hepatitis, Hodgkin's disease, Down's syndrome and an asymptomatic carrier. For the detection of HB Ag-Ab complexes a modified method of ALMEIDA was used, and for the isolation of large HB Ag particles (Dane particles) DANE'S method was employed. Electron microscopy proved to be a useful method for detecting HB Ag and the Ag.Ab complexes when the ratio of HB Ag to Ab was in the equivalence. Large aggregates of Ag-Ab complexes were frequently observed in the attacks of acute hepatitis and the recrudescences-of chronic aggressive hepatitis. The aggregates were also observed in fulminant hepatitis but the ratio of HB Ag to Ab was different from each other among 3 cases examined. The large HB Ag particles were not observed in more than half of the cases in the attacks of acute hepatitis, but appeared in the major. ity of cases in chronic aggressive hepatitis, even massively during the period with transiently elevated levels of serum glutamic pyruvic transaminase. A few large particles were also found in sera of an asymptomatic carrier, Hodgkin's disease, and Down's syndrome.