Okayama University Medical School

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.

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.

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.

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.

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.

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.

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.

Strongylidae Baird. 1853 1. Trichonema goldi Boulenger. 1916 2. T. poculatum (Looss. 1900) 3. T. longibursatum (Yorke et Macfie. 1918) 4. T. bicoronatum (Looss. 1900) 5. Bourgelatia diducta RaiJJiet. Henry et Bauche. 1919 6. Gyalocephalus capitatus Looss, 1900 7. Oesophagostomum columbianum Curtice. 1890 8. Chabertia ovina (Gmelin. 1790) RailIiet et Henry. 1909 Trichostrongylidae Leiper. 1912 9. Trichostrongylus axei (Cobb., 1879) RaiJJiet et Henry. 1909 10. T. colubriformis (Giles. 1892) Ransom. 1911 11. Haemonchus contortus (Rud., 1803) 12. Ostertagia circumcincta (Stadelmann. 1894) Ransom. 1907 13. Cooperia curticei (Giles. 1892) Ransom. 1907 14. Heligmosomum costellatum (Duj., 1845) Railliet et Henry. 1909 15. Longistriata wolgaensis Schulz. 1926

Adversive movement was first reported in 1870 by Fritsch and Hitzig when they said that when gyrus Sylvii was stimulated electrically the eyes moved to the opposite side of the stimulation. Vogt and Foerster made a detailed report on adversive movement, which is a rotary motion of the head, trunk and both eyes. But in their report, they did not make it clear what tracts the stimulation took. Mitsueda, who iS under Hayashi, defined the cortical area of the eye balls and the eye lids. He reported that they were of the extrapyramidal kind. Russel reported that when the cerebellum was stimulated the eye balls moved to the side of the stimulation, but did not say anything about adveraive movements which concerned the movement of the head and the trunk. Therefore, to ascertain the center of adversive movement and its tract the following experiments were performed. For stimulation electrical ones and chemical ones using metrazol (cardiazol) were used. As Ishizuka, who is under Hayashi, has proved excitement is only seen when cardiazol is injected among the nerve cells at a certain concentration, and it is not seen when injected among the nerve fibres.

Increase of capillary permeability is the chief symptomatic reaction of various pathologic states, especially that of localized inflammation, and this is the characteristic pharmacological properties of histamine at a far smaller concentration than that of any other chemical substances (Lewis, 1927; Crammer and Hele, 1944). There are numerous observations as to the diminishing effect of antihistamines on the flare and wheal caused by histamine and the inhibition by antihistamines of localized accumulation of intravenously injected dyes, such as trypan blue, referable to intradermal injection of histamine (for refs. cf. Loew, 1947; Fe£nberg et al., 1950). As for the inhibition of capillary permeability by antihistamines, some maintain that this action is limited to the case where such permeability has been increased by histamine (Wells, Morris and Dragstedt, 1946; Netter, 1947; Rigdon, 1949), but no single and decisive conclusion can yet be given.

Recently Kagawa found out that when the enzyme (papayo tin) substrate (gelatin) mixture had been previously irradiated with X-rays the amount of decomposition products varied in comparison with the unirradiated mixture. He thought that such a quantitative difference of fermentative products might be due to the influence of X-rays on the enzyme action. Therefore the author attempted, in the first place, to determine the optimum dose of X-rays in the proteolytic action of papayotin; in the second place, the relation between the effects of activator and inhibitor respectively and of X-rays; and finally the activity of papayotin after the papayotin substrate mixture as well as after the substrate in the mixture was irradiated.

March of spasm in epileptic convulsions was first observed by Tackson in 1863, when he said that in certain epileptic convulsions there is a phenomenon, where the convulsion starts from a certain muscle group and gradually spreads to other muscle groups. He called this, "march of spasm" and reported that it spreads according to the arrangement of motor representations in Rolando's area of the cerebral cortex. Since then, many important studies concerning the cerebral motor cortex were performed and reported. Recently, when Erickson had brought out a method in measuring electroencephalographic waves, Jackson's theory has been acknowledged. In Japan, Hayashi and his school has made an extensive study on epileptic convulsion. They used nicotine, cardiazol and others as chemical stimulations and decided the conduction tract of epileptic convulsion in dogs. The characteristic part of chemical stimulation is that, the nelve cells excite themselves when it is injected directly among them in certain concentrations and do not excite themselves when performed among nerve fibers. This was proved by Ishizuka. We used this method in dogs to see what was the mechanism of this phenomenon, "march" which is seen in epileptic convulsions and what tracts they took for conduction. And as its result, we found new facts that the presence of the motor cortex was needed for the march of spasm, and the conduction tract descending from the lenticular nucleus were quite different from Hayashi and his school had previously reported.

Acuariidae Seurat, 1913 1. Cheilospirura hamulosa (Dies., 1851) Spiruridae Oerley, 1885 2. Arduenna strongylina (Rud., 1819) Railliei et Henry, 1911 Physalopteridae Leiper, 1908 3. Physaloptera tumefaciens macaci n. subsp. Strongylidae Baird, 1853 4. Oesophagostomum maurum Hung, 1926 5. Oesophagostomum dentatum (Rud , 1803) 6. Ternidens simiae n. sp. 7. Globocephalus simiae n. sp. 8. Bourgelatia didueta Railliet, Henry et Bauche, 1928 9. Stephanurus dentatus Diesing, 1839 Trichostrongylidae Leiper, 1912 10. Mecistocirrus digitatus (v. Linstow, 1906) Railliet et Henry, 1912 Filariidae Claus, 1885 11. Setaria labiatopapillosa (Aless., 1838) 12. Setaria Bernardi Railliet et Henry, 1911 Trichuridae Railliet, 1915 13. Trichuris trichiura (Linne, 1771) Stiles, 1901

Camallanidae Railliet et Henry, 1915 1. Procamallanus annulatus n. sp. 2. Procamallanus spiralis Baylis, 1923 Cucullanidae Cobbold, 1879 3. Cucullanus sigani n. sp. 4. Cucullanus armatus n. sp. 5. Cucullanus exiguus n. sp. 6. Cucullanus arii n. sp. Heterocheilidae Railliet et Henry, 1915 7. Contracaecum arii n. sp. 8. Contracaecum sp. 9. Anisakis sp. 10. Porrocaecum sp. Larval Nematodes 11. Contracaecum larvae 12. Porrocaecum larvae 13. Raphidascaris larvae

Mononuclear cells from rabbit joint fluid were studied after synovitis was induced by various means, including the intra-articular injection of bacterial endotoxin or of aggregated human gamma globulin in normal rabbits, or of HGG in rabbits previously sensitized to this material. The large majority of mononuclear cells in all groups were monocytoid rather lymphocytoid, and these cells were most readily labeled with tritiated thymidine on the first day after injection. On day 2 and 3, the numbers of labeled cells decreased, except for the animals previously sensitized with HGG, in which there was an upswing of labeling on day 3. This upswing was associated with a considerable increase in numbers of cells resembing synovial cells, and may possibly be a reflection of synovial proliferation. Macrophages loaded with engulfed polymorphonuclear cells were observed in all experimental groups, a finding which emphasizes the lack of specificity of this reaction.

In an attempt to eliminate Japanese encephalitis virus in natural surroundings, pigs having maternal antibody were given inoculation of live-attenuated Japanese encephalitis vaccine and injection of Freund's complete adjuvant simultaneously. Titer of hemoagglutination inhibiting antibodies of pigs inoculated with live attenuated vaccine and complete adjuvant, was higher than that inoculated with vaccine alone and its titer persisted.

In vitro cell transformation of human embryo cells could be induced by the DNA purified from SV 40. The result shows clearly that cell transforms a part of viral DNA into the genome. In addition, for the purpose of clarifying th~ biological differences between the normal and transformants the alteration of cell membraneous structures of transformants (hamster and mouse fibroblasts) were observed from mechanism of phagocytosis. The iron colloid particles are taken up by normal diploid fibroblasts but not by the human and hamster transformants. This fact suggests a differ~nce in the molecular arrangement of the cell membranes between the normal and transformants. In the presence of histones, however, the transformants phagocytize the colloid particles very actively. The results show that cell membranes of transformants are altered in the molecular structure r~sponsible for the surface charge. In addition, there is no remarkable quantative differences of sialic acids on the cell surfaces of non-malignant and malignant transformants so that phagocytic activity might be correlated to the alteration of molecular composition of cell membrane itself rather than of cell surfaces, i. e, sialic acids.

The incorporation of tritiated thymidine into DNA molecules was studied by electr.:m microscopic autoradiography. To make autoradiogram in electron microscopic level, DNA was extracted from rat ascites hepatoma (AH 130) cells after in vitro incubation with tritiated thymidine. Extracted DNA samples were rotary shadowed with platinum palladium and covered with emulsion. Silver grains demonstrated on autoradiogram indicated tritiated thymidine to have incorporated into DNA molecules themselves. The incorporation was further confirmed by liquid scintillation counting of TeA soluble and insoluble fractions after DNase or RNase treatment of the DNA preparations.

Ring DNA from rat liver mitochondria has been examined by circular dichroism (CD) in the region of the 225 to 320 m/~ and the followings have been clarified. The ring DNA gives a CD spectral curve somewhat different from linear DNA from nuclei, showing a big positive peak at 266 m/~ and a small negative band at 243 m!~. That is, the positive CD band of ring DNA shifted by about 7 m/~ to the shorter wavelength side from the band of the ordinary nuclear DNA, 273 m!~. Negative band appeared at the same region as that of linear DNA but reduced in depth. Heat denaturation of the ring DNA induced a red shift of the positive band, by about 4 mp., but no change in negative band. From these experimental results it has been concluded that the ring DNA has highly twisted conformation and high in G.C contents, both of which are responsible for the blue shift of the CD spectrum.

For the purpose to investigate the physiological functions of microvillus ATPase, general properties of the enzyme were studied on the microvillus membranes isolated from rabbit intestinal epithelial cells. 1) ATPase of the microvillus membranes was activated with Mg2+. Mg.ATP complex was thought to be a subStrate of the enzyme. The Michaelis constant for ATP of the ATPase was a value of 0.8 to I .0 mM. 2) The microvillus ATPase was also activated with Ca2+, but the affinity was lower than a half of that of Mg2+. 3) The optimum pH of the ATPase was about 7.8. 4) Activity of the microvillus ATPase was markedly inhibited by treating with deoxycholate (DOC), and the activity inhibited was partially restored by washing the microvillus membrane with distilled water. The structure of the membranes destroyed by treating with DOC was also partially restored by the same procedure. 5) Ultrasonic treatment also markedly destroyed the microvillus membrane and inhibited ATPase activity. Damaged ultrastructure and ATPase activity both were partially restored by treating with phospholipid, EPL. 6) Simultaneous presence of Na+ and K + stimulated scarcely the ATPase of purified microvillus membranes. 7) The microvillus ATPase was slightly activated in the presence of n-glucose. Phloridin gave little effect on the activity of the microvillus ATPase.