Okayama University Medical School

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.

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.

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. 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.

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.

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.

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.

By assuming a three-compartment model, kinetic analysis of peripheral hyroxine distributions in various organs was represented by the alues calculated on the basis of the disappearance curve of 131I-T4 radioactivity in the serum, time　dependent curve of radioactivity over the liver　and urinary excretion of 131I-T4 in attempts to clarify the kinetic distribution　of the thyroxine and the time　dependent pool size of thyroxine in each　compartment such as serum pool, liver pool, and the other pool,　As a result it has been demonstrated that pool size3 of tyroxine, in the serum, liver and the other pool are enlarged in　hyperthyroidism, while　they are decreased in　hypothyroidism in respective pools.　We have recognized the reduction in the values of the liver pool size　of 131I-thyroxine and 131I-thyroxine excretion into the bile, while the increase　of 131I-thyroxine excretion into the urine in the cases of chronic hepatitis　and cirrhosis of the liver, but the thyroxine concentration in the serum to remain within the normal level in liver diseases. As a result of the reduction　in the liver pool size of 131I-thyroxine and in its uptake into the liver,　the other pool size enlarges to compensate the reduction and the function of the other pool is elevated.

Die stark eisenhaltigen Praparate Ferrlecit, Kobaltjerrlecit, Ferrojolsan, Ferrojolcamma und Irradian wurden im Siemens Unterrichtsreaktor SUR 100 des Instituts fur Kernenergetikder Universitat Struttgart und der Technischen Universitat, Berlin mit einem NeutronenfluB von ca. 5.l06n/cm2 sec bestrahlt. Es ollte dabei untersucht werden, wie stark eisenhaltige Praparate mit diesem Reaktor aktiviert werden konnen. Es wird festgestellt, daB Eisen praktisch kaum aktiviert wird. Die Ergebnisse zeigen, daB nur das Kobattjerrlecit und das Irradian starker durch Neutrosen aktiviert werden. Die hohere Aktivitat des Kobaltjerrlecit wird auf die darin enthaltenen Elemente Mangan, Kupfer und Kobalt zuruckgefuhrt, die mit thermischen Neutronen gut aktivierbar sind. Die hohere Aktivitat des Irradian geht hauptsachlich auf β-Zerfalle zuruck. Das Phosphorisotop P-32 tragt einen groβen Anteil dazu bei. De rβ-Untergrund des Irradian bei hohen Energien, der mit einer kleinen Halbwertszeitabklingt, ist durch keines der angefuhrten Elemente im Irradian erklarbar. Der Mn-56Peak im Spektrum ist gut erkennbar, durch den β-Untergrund wird er aber zu einer hoheren Energie hin verschoben.

Two cases of multiple foreign bodies, i. e., duplex coins in the esophagus are reported. These foreign bodies were removed by peroral esophagoscopy successfully. Significance of roentgen-ray diagnosis is emphasized, and subtle and yet specific roentgenograms of duplex coins in the esophagus are illustrated.

An electron microscopic observation was made on the DNA's extracted from purified HeLa cell nuclei, mitochondria, and the whole cell, and fractionated by ethidium bromide-cesium chloride density gradient method or sucrose density gradient method. Nuclear DNA presents mainly long linear DNA derived from fragmented chromosomal DNA. In addition to this, the existence of small circular DNA molecules measuring 0.32 -1.78 μ, was confirmed. Mitochondrial DNA was mainly circular DNA, which measured 4.87 μ in the mean value of the contour lengths in the highest frequency group, and small circular DNA molecules, measuring 0.3-1.01 μ in contour length, were also found in an extremely low frequency.

A human fetal fibroblast strain, belonging to a group resistant to SV40 transformation, was transformed by SV40 through a multiple inoculation procedure. Two independently transformed cells were described in comparison with each other. The proportions of cells with the nuclei possessing V antigen were 2.9% at the 5th passage in one strain and 1.1 % at the 4th passage in another, and they declined gradually as frequent passages were repeated. The percentages of the transformed cells with V antigen-positive nuclei were, in both strains, quantitatively compatible with those of the cells with the nuclei full of virus particles in crystalline arrays, which were demonstrated by immunofluorescent studies and electron microscopy.

A non.specific esterase activity was demonstrated III the jejunum of rats by an azoindoxyl method. 1) Microvilli of the jejunal epithelial cells were remarkably stained in non-frozen specimens and feebly in frozen specimens. 2) The other cytoplasmic structures, i. e. mitochondria, endoplasmic reticulum, nuclear envelope, plasma membrane and multivesicular body showed a positive reaction product in frozen sections but not in non-frozen blocks.

Die Autoren haben 48 Patienten mit chronischer Koronarinsuffizienz mit Nitrolingual Retard Kapseln bettlagerig oder ambulant behandelt. Nach ihren Ergebnissen kann man sagen, dass Nitroglycerin in einer Langzeitform als Nitrolingual Retard Kapsel den modernen Erfordenissen der Therapie der Koronarinsufflzienz entspricht, da damit iiber einem langeren Zeitraum hinweg gleichmassig Blutspiegelwerte erzielt werden konnen. So hat man einen guten therapeutischen Erfolg, dazu einen Riickgang der Anfallhaufigkeit und eine erhohte Belastbarkeit erreicht. Der Wirkstoff hat iiber einem Zeitraum von 10-12 Stunden sich konti. nuirlich diffundiert bzw. klinisch gewirkt. Bei der Einnahme von Nitro. lingual Retard sind keine Kopfschmerzen aufgetreten. Die Vertraglichkeit des Praparateserwies sich als hut.

For the purpose to reveal the substance as the source of offensive odor of the fish from the sea facing petroleum and petrochemical industries, analyses have been made on the sea water, industrial wastes, offensive odor fish meat and the eels kept in the sea or industrial waste for a certain period, by means of gas chromatography, infrared, and ultraviolet absorption spectrophotometries and mass-spectrometry. Observations revealed toluene as a possible main source responsible for the bad smelling of the fish. Finally, by analysing the meat extract of the fish kept in the sea water containing toluene and by the same smell as that of the offensive odor fish from the off-shore of the industries, it has been definitely confirmed that toluene is the very substance that imparts the offensive odor to fish. The activated sludge process proved to be an effective method to remove toluene from the industrial wastes. A discussion was made on the aliphatic carbohydrate as the possible source of the offensive odor of fish.

Epidemiological study on the prevalence of SMON III Ibara City, Yoshii Town and Yubara Town in Okayama Prefecture was conducted and the following results were obtained. "Epidemization precession" (Die Prazession der Durchseuchung), described by Rudder which shows that ages common to this disease is shifting to younger generation as incidence rate increases, in the endemic regions was recognized in Ibara City and Yubara Town. The mode of the distribution of the interval between the primary and secondary patients in a family suggests that it corresponds to the incubation period. Distribution of the patients to the size of family is not adaptable to binomial model, but adaptable to chain binomial model, and the intra-household transmitted rate is 5 per cent, and the true intra-household transmitted rate is about two times as high as extra-household transmitted rate by the modified chain binomial model. Intra-hospital incidence among SMON patients and non-SMON patients as well as medical workers were observed at the time when the number of new patients increased in a hospital. In SMON patients, the rate of appendectomized ones was higher than that of non-SMON patients, suggesting that gastrointestinal tract has some relationship to the development of SMON disease.

We applied unidirectional MLC test to skin allografts in dogs, and examined the correlation between the strength of the MLC reaction and the survival time of skin allografts. As a result it was found that the skin allografts was rejected within 10 days when the rate of blastformation was more than 18 %. In contrast, the skin graft survived over 13 days when the rate was less than 15 %. The rate of blastformation was inversely correlated with the median survival time of skin allograft.

Morphological comparison at colonial level was made on a series of established liver cell lines derived from rats fed 4-dimethylaminoazo-benzene (DAB) for various periods of days for the purpose of elucidating more accurately the differences in morphology and growth patterns among these cell lines. Colonies of each cell line produced by the single cell plating technique were compared with regard to colony size, density and piling-up of cells, atypism and pleomorphism of cells, and the migration of cells from colonies. Plating efficiency of each cell line was also compared. The cultured rat liver cells obtained from those rats fed DAB for a longer period of days showed higher plating efficiency, and increased the incidence of large-sized, dense, and piled-up colonies, of colonies consisted of cells having nuclear atypism and pleomorphism, and of irregularly margined colonies with migrating cells. The correlation between the present results and the process of DABcarcinogenesis is discussed.

The first successful electron-microscopic observation of a virus isolated from a patient with SMON was performed. The morphological and developmental characteristics of this virus suggests that this type of virus has not been isolated from humans. Hence, it is considered that the virus observed is of a new type and presumably the causative agent of SMON. The author wishes to express his profound thanks to Prof. TADASHI OFUJI for painstaking proof reading of the manuscript and also acknowledgement is due to Mr. NOBUO HAYASHI, Mr. NOBORU SAIHARA, Mr. TAKASHI NAKAMURA and Miss TOSHIYO OMIZU for their technical assistance of electron microscopy.

As to trial toward the elimination of Japanese encephalitis virus in natural surroundings, pigs received inoculation of inactivated Japanese encephalitis vaccine supplemented with complete Freund's adjuvant twice at one-week interval. Effect of adjuvant supplement on the magnitude of antibody and also prevention of viremia caused by natural infection by antibody induced with vaccine were investigated. The results of this study are summarized as follows. 1. In the group of pigs inoculated with vaccine containing adjuvant, titer of hemoagglutination inhibiting and neutralizing antibodies was higher than those inoculated with vaccine alone and their high titer persisted. 2. With respect to natural infection of pigs, on August 22 when the pigs were thought to have been infected, there was observed a rise in antibody titers. And on antibody formed in those pigs inoculated with vaccine with or without adjuvant proved to be all 2-ME resistant type, whereas the antibodies produced in control group were 2-ME sensitive antibody. 3. Viremia was detected in the blood of pigs naturally infected, but it was not demonstrated pigs inoculated with vaccine supplemented with adjuvant or without adjuvant. The virus of pig blood which was inoculated into suckling mouse brain and was separated after low suckling passage mouse was supposed to be JaGAr strain from optimum hydrogen ion concentration of its hemoagglutination reaction. 4. Effect of vaccination on antibody response of pigs having maternal antibody was not recognized.