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With the purpose to clarified the mode of localization and mechanisms of activation of ATPase in the mitochondrial membrane, analyses were made on the properties of mitochondrial ATPase from the structural and functional aspects. The activation of ATPase by DNP and Mg++ and the oligomycin sensitivity were investigated in a series of inner membrane fragment samples obtained by ultrasonic irradiation and those samples obtained in the processes of isolation and purification of ATPase from rat liver mitochondria and beef heart mitochondria in parallel with electron microscope observations. As a result it has been found that the membrane fragments obtained from rat liver and beef heart mitochondria by ultrasonication exhibited high respiratory activity and unmasked ATPase activity which was charac· terized by remarkable stimulation by Mg++ and inhibition by oligomycin and azide. Therefore, mitochondrial ATPase seems to be bound fairly closely to the inner mitochondrial membrane. In the membrane fragments prepared by ultrasonication of intact mitochondria, ATPase activity was stimulated by DNP, but in the supernatant fractions was not. On the other hand, the supernatant fraction obtained from BHM and inner membrane fragments by severe sonication exhibits a marked ATPase activity and the activity incresed in each step of the purification on the treatments with acid, protamine and heat. Especially in the case of membrane fragments the protamine treatment can be omitted. Electron microscope observation of the fractions in each step of the purification proved the head pieces to be ATPase. The ATPase activity of solubilized head pieces is insensitive to oligo. mycin and coincides with the soluble ATPase of PULLMAN etat. (8) in the points of its cold labile property and optimum pH, but it shown no accele. ration of ATPase activity by DNP.

For the purpose to obtain the information of the mechanism of protein uptake by the tumor cells, some cytochemical and electron microscopic observations were carried out on Ehrlich ascites tumor cells incubated with horseradish peroxidase (basic hemoprotein, molecular weight approximately 40,000) in vitro. In the earlier periods of the incubation peroxidase was found to be adsorbed on some area of surface of the tumor cells forming a thin protein layer, where an active pseudopodia formation was observed. With the lapse of time, the protein was taken in the deep cytoplasm by the infoldings of the cell membrane and accumulated in the cytoplasmic vesicles having limiting membrane. Concerning the accumulation of the protein into the vesicles, small tubular structures in the cytoplasm connecting the cell surface and the vesicles, were considered to participate in the intracellular transportation of peroxidase taken up. In cold environment (2°C), the formation of pseudopodia and deep inward infoldings of the cell membrane was inhibited and simultaneously the uptake of peroxidase stopped. Iodoacetate and sodium fluoride also effected to suppress the pseudopodia and infoldings formation moderetely, as well as uptake of peroxidase, though they did not stop completely. These facts have indicated that horseradish peroxdase is taken up by Ehrlich ascites tuimor cells through pinocytosis which involves energy-requiring process dependent upon glycolytic metabolism of the tumor cells.

For the purpose to reveal the changes in the metabolism of erythroblast in varied specialization stages the author observed the Feulgen DNA level of rabbit erythroblasts by microspectrophotometry. Observations were made on normal and anemic animals, and those receiving a mass red cell transfusion at the recovery stage of anemia where the early denucleation is stimulated. Observations have revealed that in normal erythropoiesis the DNA contents are kept at n to 2 n level from the proerythroblast to late basophilic stage, but in later stages, polychromatic and orthochromatic, DNA level per cell decreases gradually with advance of the cell specialization reaching the minimum level, nearly haploid level, at orthochromatic stage where most cells are believed to be denucleated. In blood depleted animals nearly the same pattern of DNA level was observed in connection with erythroid specialization as that in normal animal, except a relatively high DNA level in the later specialization. In the cases of the hemolytic anemia a similar tendency has been observed but the minimum level of DNA remains at a higher level, hypo-diploid level, in poly- and orthochromatic stages. Twenty-four hours after the mass red cell transfusion by which severe anemia has been recovered to the original level within one hour, the pattern of the DNA level of the erythroblast returns to the normal one showing a very low DNA level at the polyand the orthochromatic stages. The data indicate that the DNA synthesis of erythroblast kept at n to 2 n levels until the late basophilic stage begins to decline at polychromatic stage and reaches nearly haploid level at orthochromatic stage, but in active hemopoiesis the DNA synthesis is stimulated and the DNA contents are kept at a high level even in the late specialization stages, showing no relation between the denucleation and the low DNA level.

Localization of IgM and IgG sypylitic antibodies in the sera of patients and the experimental syphylitic rabbits was examined by the gel filtration on Sephadex G.200 column. I) In the case of late syphylitic patients; OGATA test-reactive antibodies were contained in IgM and IgG fractions. On the other hand, RPCF test-reactive antibody was contained only in IgG fraction. This discrepancy may be due to the difference in antigens; Cardiolipin.resicin and T. P. Reiter protein. 2) In the case of the experimental syphylitic rabbits; The results were as follows. a) Variation in the level of the titer. The peaks of the titer were seen 3-4 weeks after inoculation of T. P. Nichols by OGATA test, VDRL test and RPCF test, thereafter titers decreased. On the other hand, the titer kept on rising up to 2 months and maintained high level during the periods of 3, 4 and 5 months by TPHA test. b)Transformation of antibody from IgM to IgG. Transformation of antibody from IgM to IgG was seen 3-4 weeks after inoculation by all four tests; OGATA test, VDRL test, RPCF test and TPHA test, and such a transformation was completed 3 months after inoculation.

1) Normal karyotype of Donryu strain rat was determined according to the classification of KURITA et al. (8). Namely, the number of chromosomes was 42 in diploid cells, and chromosomes were divided into 3 groups (A, B and C) according to the position of centromere. A.group was consisted of 7 pairs of metacentric chromosomes, B-group 4 pairs of sub· meta.subtelocentrics and C.group 10 pairs of telocentrics and Y. 2) Among all chromosome pairs a pair of the longest telocentric chromosomes (C.l), 4 pairs of all the B.group, and the Y chromosome were recognizable. 3) The presence of polymorphism was demonstrated in the smallest submetacentric chromosomes (BA),: namely, (I) a homologous submeta. centric pair, (II) a homologous subtelocentric pair and (III) a heteromor. phic submeta and subtelocentric pair which seemed to be a hybrid from (I) and (II). To distinguish the polymorphism in their genotype from phenotype was impossible. 4) Animals with type III B·4 chromosomes were produced by type I and type II animals. 5) By checking the chromosomes of the inbred Donryu strain rats maintained over 40 generations by brother.sister mating at Nihon Rat Co., polymorphism in BA chromosomes was also recognized.

Female Cb mice weighing 20-23 g were exposed to 800 ppm (in average) of 1, I, 2, 2.tetrachloroethane for 3 hours. Both triglyceride and phospholipid in the liver and plasma were determined at varying times after the exposure. On the other hand, there were observed the ultraviolet absorption spectra of microwmallipids in the liver at 90 minutes after the 1, 1,2, 2-tetrachloroethane or the carbon tetrachloride exposure. The results thus obtained are summarized as follows: 1. The increase of hepatic triglyceride contents attained the maximum level in the period between 20 and 25 hours after the exposure and declined to the initial levels at 90 hours later. 2. The plasma triglyceride levels decreased until 25 hours after the exposure, then tended to increase significantly and were much higher than the control levels in the period between 70 and 90 hours later. 3. Both liver and plasma phospholipid levels decreased gradually up to 25 hours after the exposure, then slowly recovered with almost the same rate of increase. 4. It was suggested that the inhalation of the above vapors induced a little change in microsomal lipids in the liver.

For the purpose of forecasting the prevalence ofJapanese encephalitis in Japan, we tried to find out the correlation of factors between median and mode dates of epidemic time curve of prevalence on one hand, and average atmospheric temperatures of prefectures in June and July (T6,7 in short) (X¹), the time when HI reaction of swine became positive to the degree of 50 per cent (D. pos. swine in short) (X²), the latitude (x³) and longitude (x4) in respective prefectures (in 1965 and 1967). On the other we also estimated the median and mode dates of this epidemic curve of the prevalence in 1968 and 1969, from the regression equation of one variable and multiple regression equation from the above factors using an electronic computer. The usefulness of adding factors concerned with mosquitoes to the above four factors is proven by the accuracy of estimation. And the following results were obtained. 1) Phenomenally speaking, the prevalence of Japanese encephalitis follows the principle of "advancing of prevalence towards the north and east" and essentially speaking, it depends upon high atmospheric temperature and the outbreak of many hazardous mosquitoes by the high atmospheric temperature. 2) To estimate median date (y) and mode rate (z) of the epidemic time curve of the prevalence, we can use the next equations; The regression equations to estimate y and z from T 6,7(X) are as follows. y = - 3. 75X¹ + 144.47 σ = 12.4.·. [1] z = - 3. 80X¹ + 157 .26 σ = 14.9.. · [1]' The regression equation from D. pos. swine (X²) are as follows. y = 0. 68X² + 31. 82 σ = 9.2· .. [2] z=0. 76X² +40. 71 σ= 12.0 .. · [2]' The multiple regression equation from T6 ,7 and D. pos. swme are as follows. y = -1. 07X¹ +0 .62x² +59. 37 σ= 9.7 ... [3] z= -0. 79x¹ +0. 71x² +61.02 σ= 12.0· .. [3]' The multiple regression equations from T 6•7, D. pos. swine, latitude and longitude are as follows Y= -1.01x¹ +0.58x² -0.26x³+0 .37x4 + 18.50 σ= 9.8･･･ [4] z = -0. 32x¹ +0. 52x² +2 .05x³ +0 .54x4 -87. 81 σ= 11.8 [4]' 3) We Obtained the estimated value of median date in 17 prefectures in Kyushu, Chugoku, Shikoku, Kinki and Kanto provinces in 1968 and in 13 prefectures in 1969 from [l] or [2] or [3] or [4] equation. Nine prefectures out of 17 by [l], 12 prefectures by [2], 13 by [3J and [4] in 1968. [4] could be estimated with about 10 days error or less. And in 1969, 9 out of 13 by [3] and 7 out of 13 by [4] could be accurately esti· mated. The estimation by the multiple regression equation using many factors is most useful for the calculation. 4) The time when the number of patients increases at maximum can be pointed out by the lower limit of prediction region obtained from data in each prefecture. And the lower limit was the estimated median value minus about 20 days by [1] and about 16 days by [2] or [3] or [4] under the next condition; α = 0. 1, N= 75. 5) The mode dates in 17 prefectures out of 19 were estimated by [1]', [2]', [3]' and [4]'. 12 prefectures out of 17 by [1]', 7 by [2]', 10 by [3]' and 13 by [4]' could be estimated with about 12 days error or less in 1968 and 9 out of 13 was correctly estimated by [3]' and [4]' in 1969. The estimation by the regression line of one factor was s~mewhat different from each other, but when multiple regression line of four factors was used the estimation became more correct. Judging from these results, it is adequate to use the multiple regression equation of [4] and [4]' when we want to forecast the median date or mode date ofJapanese encephalitis time cure. 6) In the case of adding two factors concerned with mosquitoes to T6,7 (X¹), D. pos. swine (x²), latitude (x³), longitude (x4), multiple regression equations become as follows. y= -1.46x¹+0.14X²+0.068x5+89.03 σ= 6.9.. ·[5] z= -3. 29x¹+0 .13x²-0. 010x5+ 143.63 σ= 18.6··· [5]' y=-4.20x¹+0.35x²+0.29x6 + 53.70 σ= 4.2 .. ·[6] z=-2.56x¹-0.0lx²-0.02x6 +128.96 σ=11.4 [6]' y= 4.76x¹+0.41x²+0.13x5+0.22x6-72.78 σ= 4.5 [7] z = - 2. l0x¹ + 0. 05x²+ 0. 11 x5 - 0. 08x6+ 113.4 σ= 10. 7.. · [7]' where x5 is the time when the number of mosquitoes (C. T. collected by light trap reached the maximum and X6 is the time when hazardous mosq uitoes were dected. In the case of median date, 5 prefectures out of 6 prefectures by [5], 2 out of 6 by [6] and 2 out of 5 by [7], and in the case of mode date, 5 out of 6 by [5]', 4 out of 5 by [6]' and 4 out of 5 by [7]' could be accurately estimated in 1969.

1. Twenty.one patients with liver diseases were studied for their urinary mehylmalonic acid excretion after a valine load by means of an improved thin layer chromatography. 2. Methylmalonic acid positive cases were found in four out of the ten patients with cirrhosis of the liver, all four with cirrhosis and diabetes mellitus, and none with acute hepatitis of icteric phase. No apparent correlation was found between the methylmalonic acid excretion and the extent of hepatic damage. 3. A large amount of methylmalonic acid found in the case (S. I.) with cirrhosis of the liver and diabetes mellitus after the valine load was not corrected by cyanocobalamin but by DBCC, suggesting an impaired transformation from cyanocobalamin to DBCC. However, the nature of the impairment remains unknown.

The correlation of infectivity to the length of SV 40 DNA by dilute and undiluted passage was described. DNA was extracted from purified virions by Marmur's method, and propagation of virus was done using VERO cells. The infectivity of SV 40 (simian virus 40) ran parallel with the length of its DNA. Undiluted passage caused shortening of DNA and decrease in infectivity, but when these undiluted group was passaged dilutely, length of DNA approached the original length and the infectivity recovered. In undiluted passage group small circular DNAs under 1.0,11_, so far not reported in the SV 40 DNA, were found in low frequency. Replicating form and dimers were also noted from virions and the nuclei of SV 40 infected VERO cells.

The effect of thioinosine on antibody response of immunized rabbits has been studied. When given simultaneously with the antigen, thioinosine profoundly suppresses the formation of humoral antibody.

1. The mixed lymphocyte culture test where live lymphocytes of female mice are cultured with supersonicated cell homogenate of isogeneic male mice does not reflect the difference in H-Y histocompatibility antigen alone. 2. When non-H-2 antigens are cumulated in various combinations, the rate of blastformation becomes greater than the combination of C3H ♀ themselves. 3. When non-H-2 antigens and H-Y antigens are added in various combinations, the rate of blastformation becomes significantly higher than in the combination of C3H ♀ themselves.

Die Autoren haben uber ihre Erfahrungen mit dem neuen Glutisal- Versuchspraparat - einer Kombination von Salicylamid und Dimethyl- aminophenyldimethylpyrazolon im Zusammenhang mit der Behandlung von 350 Patienten berichtet. Es wurden vor und wahrend der Behandlung instrumentell die Schmerzempfindlichkeit, die Funktion der quergestreiften Muskulatur (Tonus und Kontraktionsamplitude), Kreislauf, Nerven- und Gelenkfunk· tion mit dem Myotonometer und Angioneuromyographen (nach SZIRMAI), Oszillometer, Oszillographen, Angiometer registriert. Es wurde die Harn- saureausscheidung bei Gichtfallen registriert. Die Autoren fanden, daβ das Praparat die Erkaltungen, verschiedene rheumatische Erkrankungen sowie Gicht gunstig beeinflusst. Die Vertra- glichkeit war sehr gut.

For the purpose to clarify the distribution of DNA in mouse ascites sarcoma cells (SR-C3H) induced by Rous sarcoma virus (Schmidt-Ruppin strain), quantitative assays were carried out by SCHMIDT-THANNHAUSERSCHNEIDER'S method using subcellular fractions isolated from SR-C3H cells and C3H mouse liver as a control tissue, and simultaneously electron microscopic observations were conducted with the rotary shadowed preparations of the SDS-phenol extracted nucleic acids by the protein monolayer technique. The results are briefly summarized as follows. 1. The RNA/DNA ratios in SR-C3H cells and liver cells were 2.3 and 3.7, while those in nuclear fraction of SR-C3H cells and liver cells were 0.34 and O. 56, respectively. The electron micrographs of nuclear nucleic acids revealed a DNA-RNA complex-like structure. 2. DNA and RNA contents of SR-C3H mitochondria were found to be 3.1 and 24 fl-g per mg of protein, respectiVely, which proved to be greater than those of liver mitochondria. The mean values of the contour length of circular DNA molecules in highest frequency group observed in the electron micrographs were 4.88 μ. in SR-C3H mitochondria and 5.08 μ. in mouse liver mitochondria. There could be observed circular molecules of duplicated-length in both mitochondrial DNA's and small circular molecules in SR-C3H mitochondrial DNA. 3. In the microsomal and supernatant fractions of SR-C3H cells and mouse liver cells, the ratios of DNA to RNA gave several percent by chemical analysis and this percentage was particularly high in the supernatant of SR-C3H cells. On the other hand, in the electron micrographs, the fibrous structure was significantly recovered in the supernatant nucleic acids of SR-C3H cells, but with difficulty in the other three fractions. This fibrous structure measured 1.13 μ in the mean value of the length and was considered to be DNA as it readily disappeared after the treat· ment with DNase.

1. Marsupialization was performed on 23 cases of cystic lesions of the jaw at the Department of Oral Surgery, Okayama University Hospital in the recent five years. 2. These patients were divided into 4 groups according to their age; namely, group A of those under IS years old, group B of those between 15 and 30 years old, group C of those between 31 and 60 years old, and group D of those over 61 years old, and the results of postoperative findings were compared with those of preoperative ones. 3. In group A of the four groups the most favorable results were obtained after marsupialization and reduction of the tumor was remarkable, even in a case of ameloblastoma, as compared with groups B, C and D. 4. Cure of lesions took somewhat a longer period of time in group B than in group A, but all the cystic lesions were reduced favorably after operation. 5. The reduction in cysts in groups C and D was markedly slower when compared with that in groups A and B, but the marsupialization surgery seems to be desirable in some cases.

The rate of transport of blood glutamic acid into the brain and the rate of metabolic conversion of the amino acid in the brain were derived by the use of the brain perfution method in vivo and in situ with [D.HC] ·Lglutamic acid. The net uptake of glutamic acid by the brain was observed. Most of the radioactivity released from the brain into the cerebral venous blood was found to consist of glutamine. Small but significant amounts of output as radioactive GSH and CO2 were also found. Glutamic acid transport and its rate of metabolism were lowered in the glucose. free condition. The size of the compartment of the small glutamic acid pool, which was related closely to the blood glutamic acid, and that of the large glutamic acid pool, which was related closely to the blood glucose, were calculated and compared with each other.

Antigenicity of clozapine was studied in rabbits, comparing with that of chlorpromazine as control. The results indicate that chlorpromazine produces antibody in rabbit as revealed by passive hemagglutination test, giving the titer of 1 : 2, 000 or higher in all the five cases observed, though specific precipitin lines has not been obtained and PCA test proves to be negative. Clozapine failed to produce anti.clozapine antibody giving negative passive hemagglutination test, passive cutaneous anaphylaxis and precipitin reaction, in all forms tested. Some remarks were made on the possible close relation between the antigenicity of the drug and its affinity to protein.

For the purpose to reveal the relation between cell death and nuclear stainability by supravital staining with basic dyes observations have been made on the cells of bone marrow, peripheral blood and lymph node from anemic and non-anemic rabbit, rat, mouse and chicken, and thymus from young mouse. The cells were stained supravitally in blood serum, isotonic saline, calcium chloride and sucrose solutions with the dyes; brilliant cresyl blue (B. C. B), Nile blue (N. B.), neutral red (N. R.), Janus green (J.G.) and eosin (E.). The following results were obtained: 1. In the presence of blood serum all the living cell nuclei observed were not stained supravitally, except some mature erythroblasts and nucleated red cells. 2. In isotonic saline, CaCl2 and MgCl2 solutions all the erythroid cell nuclei were stained deep by B. C. B., N. B., N. R., slightly by J. G. but not by E. In stainability the younger the cell is the deeper in its nuclear staining. The nuclei of other cell strains were not stained. 3. In isotonic sucrose the nuclei of mature granulocyte were also stained by B. C. B. and N. B. but not by other dyes. The nuclei of lymphoid cells and myeloid cells appeared pale without being stained by any dyes. The nuclei of erythroblasts in sucrose solution were stained deeper with B. C. B. and N. B. than those in isotonic saline. The differences between supravital stainability of the nuclei among the cells belonging to different strain and among those of the same strain but in different maturation stage and the nuclear staining after cell death have been discussed from the possible dissociation of DNA from histone.

Antigenicity of the peptide of ribosomal digest in Ehrlich ascites tumor was studied. The peptide was purified by DEAE Sephadex A-50 column chromatography. The peptide was electrophoretically basic, single, and 1.32 S20w sedimentation coefficient with poor content of tyrosine and phenylalanine. The maximum absorbancy was at 267 mμ. Mice and rabbits were immunized with the mixture of the purified peptide with Freund's complete adjuvant. The inhibitory effect of immune γ-globulin on the tumor growth was demonstrated in vitro cultured JTC-11 cells. A single precipitin line was observed between rabbit antiserum and tumor cell extract of Ehrlich ascites cells in ouchterlony double diffusion chamber and immunoelectrophoresis. The sedimentation coefficient of the effective fraction in immune-serum was 17 S20w. The precipitin line was observed at β2-γ region in immunoelectrophoresis.

A unique low density lipoprotein was obtained from the tumor transplanted with a cultured cell line of Ehrlich ascites tumor, JTC-ll cell. The tumor low density lipoprotein electrophoretically migrated as a single band, and the mobility was different from that of other organs. The chemical composition of lipid, cholesterol and phospholipids in tumor low density lipoprotein were characteristic. The flotation rate was Sf 5.9, and thus the molecular weight was estimated to be about 130 x 104. The inhibitory effect on tumor growth of the immune serum was most elevated at 25th day after the intraperitoneal administration of tumor low density lipoprotein. The main fraction effective for inhibition of tumor growth existed in γ-globulin.