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It is well known that eosinophils are involved in tyrosine nitration. In this study, we evaluated tyrosine nitration by rat eosinophils isolated from peritoneal fl uid and constituent eosinophils in the stomach. Rat peritoneal eosinophils activated with 1 μM phorbol myristate acetate (PMA) and 50 μM NO2 ﾝ showed immunostaining for nitrotyrosine only in smaller cells, despite the fact that eosinophils are capable of producing superoxide (O2·ﾝ). Free tyrosine nitrating capacity after incubation with PMA and NO2 ﾝ was 4-fold higher in eosinophils than in neutrophils. Catalase and ｸ- and ｺ -tocopherol inhibited free tyrosine nitration by reactive nitrogen species from eosinophils but not that by peroxynitrite. Superoxide dismutase augmented free tyrosine nitration by activated eosinophils and peroxynitrite. The concentration of nitric oxide released from eosinophils was relatively low (0.32 μM/106 cells/h) and did not contribute to the formation of nitrotyrosine. On the other hand, most constituent eosinophils constituent in the rat stomach stimulated by PMA and NO2 ﾝ showed tyrosine nitration capacity. These results suggest that intact cells other than apoptotic-like eosinophils eluted in the intraperitoneal cavity could not generate reactive species responsible for nitration by a peroxidase-dependent mechanism. In contrast, normal eosinophils in the stomach were capable of nitration, suggesting that the characteristics of eosinophils in gastric mucosa are diff erent from those eluted in the peritoneal cavity.

To date, many techniques have been reported for vaginoplasty in male-to-female trans-sexual (MTFTS) patients, such as the use of a rectum transfer, a penile-scrotal flap and a reversed penile flap. However, none of these procedures is without its disadvantages. We developed a newly kind of flap for vaginoplasty, the M-shaped perineo-scrotal flap (M-shaped flap), using skin from both sides of the scrotum, shorn of hair by preoperative laser treatment. We applied this new type of flap in 7 MTFTS patients between January 2006 and January 2007. None of the flaps developed necrosis, and the patients could engage in sexual activity within 3 months of the operation. The M-shaped flap has numerous advantages: it can be elevated safely while retaining good vascularity, it provides for the construction of a sufficient deep vagina without a skin graft, the size of the flap is not influenced entirely by the length of the penis, and it utilizes skin from both sides of the scrotal area, which is usually excised.

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.

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.

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.

For purpose to study specificity in the growth inhibition effect of sensitized lymph-node cells on target cells, the regional lymph-node cells obtained from the truly isologous mouse previously inoculated with A strain cells (derived from C3H mouse mammary cancer) were cultured with A cells in various ways, and obtained the following results. 1. Those regional lymph-node cells from the isologous mice transplanted with the skin of C3H mouse or MC-induced sarcoma do not inhibit the growth of A cells in tissue culture. 2. The regional lymph-node cells from the mice positive to tuberculin test also do not inhibit the growth of A cells in tissue culture. 3. The regional axillary lymph-node cells (C3H anti-A strain cells) inhibit the proliferation of M cells from Cb mouse mammasy cancer and JTC-ll cells from Ehrlich ascites tumor as well as A cells. However, these axillary lymphnode cells do not inhibit the growth of AH-66F cells from rat DAB hepatoma, Hela-S3 cells from human uterine cancer and L cells from subcutaneous connective tissue of C3H mouse. From these results it is assumed that the sensitized regional lymph- node cells act specifically on cancer antigen.

Detailed morphologic characteristics of type C virus particles observed in an X-ray-induced C58 mammary tumor and its transplants have been described. The particles are round and 75 to 100 mμ in diameter, containing an electrondense nucleoid 60 to 70 mμ in diameter. By the negative staining, they do not show obvious spines. Two abnormal types of particles, i. e. cylindrical and aberrant forms have been observed.

Cultured rat liver cells which were cloned from a single cell were transformed into malignant cells by a chemical carcinogen, 4-dimethylaminoazobenzene (DAB). The DAB-transformed cells produced tumors when back-transplanted into new-born rats but the carcinogen-untreated control cells did not. Characteristics of the transformed liver cells were compared to those of DAB-untreated control cells in regard to the morphology, the consumption of DAB from the culture medium by the cells, the incorporation of 3H.DAB into the cells, and the aggregate.forming ability of the cells in rotation culture. The results showed that no significant parameter of malig. nant transformation in culture was detectable except the tumorigenicity of the transformed cells upon the inoculation into animals.

Using a freeze-etching method, the ultrastructure of cell surface of gram-positive cocci was studied by digesting cell wall with lytic enzyme. In M. lysodeikticus, the cell surface revealed a very simplified ultrastructure, i. e. a single cell wall layer and a single plasma membrane layer. On the contrary, the cell surface of S. aureus exhibited a unique structure composed of two cell wall layers and a single ploasma membrane layer. The wall layers were constituted of 160 -180 A particle layer (CWl) which was unsusceptible to the L-ll enzyme and amorphous layer (CW2) which was susceptible. These results suggested that 160-180 A particles in CWl consisted mainly of the teichoic acid.

To test the hypothesis that the endothelium-derived relaxing factor (EDRF) contributes to coronary vasodilation induced by myocardial ischemia, we examined the effect of NG-nitro-L-arginine (a potent and selective inhibitor of EDRF release) on the coronary reactive hyperemic response in the open-chest dogs. Intracoronary infusion of NG-nitro-L-arginine at a coronary plasma concentration of 5 x 10(-5) M had no effect on hemodynamics and myocardial oxygen metabolism, but attenuated repayment of the flow debt by an average of 20.4% and 20.0% following coronary occlusion for 10 sec and 20 sec, respectively. Concomitant infusion of NG-nitro-L-arginine at the same concentration and 8-phenyltheophylline (a potent adenosine receptor blocker) at a coronary plasma concentration of 10(-5) M further attenuated flow debt repayment following 10 sec and 20 sec of coronary occlusion by 47.7 and 59.4%, respectively. These results indicate that EDRF plays a significant role in the coronary reactive hyperemic response and may cause vasodilation independently of adenosine-mediated vasodilation following coronary occlusion.

As has been well established, reticulocytes (RC) synthesize the species specific protein, globin, actively for about 24 hours or more till the time of their complete maturation1,2,3. This will be possible only in the presence of messenger RNA (m-RNA)4,5. Since the splendid hypothesis of m-RNA proposed by JACOB and MONOD6 for explaining the mechanism of the transfer of genetic information from nucleus to cytoplasm, it has largely been accepted through the numerous observations that followed7,8,9,10. However, the m-RNA hypothesis, which has been deduced by observing the protein synthesis in E. Coli, includes the meaning of labile RNA which is incessantly decomposed and newly synthesized to compensate the rapid degradation. As m-RNA cannot be synthesized in RC which have no detectable DNA, it has been supposed that the m-RNA of RC should be considerably stablell,12,13. Even in the denucleated cells, however, the RNA synthesis might be possible because Borsook reported the positive RNA synthesis of RC14, and this result has recently been reconfirmed by BURNY15.

1. The studies of structure and function of the plasma membranes of cancer cells is extremely important for the elucidation of specificity of phenotypes of cancer cells. In order to bring this subject to light, plasma membranes, mitochondria, microsomes and nuclei have been isolated from the AH 130 ascites carcinoma cells and rat liver cells. The electron cytochemical observations and biochemical assays of M g²+-Na+-K+-ATPase, ADPase, AMPase, and β-glycerophosphatase activities have been carried out before and after the fixation with glutaraldehyde. 2. M g²+-ATPase and Mg²+-N a +-K +-ATPase are present in the isolated plasma membranes, mitochondria and microsomes in both AH 130 cells and rat liver cells. ADPase and AMPase of the mitochondria and microsomes show far lower activities than those of the corresponding enzymes found in rat liver plasma membrane. ADPase and AMPase of AH 130 cell fraction exhibit activity much lower or zero. Generally, enzymatic activity of the AH 130 cell fraction is much lower than that of rat liver cell fraction. 3. Mg²+-Na+-K+-ATPase is completely abolished by 5% glutaraldehyde fixation while it shows less effect on Mg²+-ATPase in the plasma membrane. ADPase and AMPase activities of the mitochondria and microsomes are completely inhibited by glutaraldehyde fixation. AMPase of the plasma membrane of rat liver is completely abolished while ADPase activity is not affected in any way. 4. Only Mg²+-ATPase can be demonstrated electron cytochemically. Cytochemical reaction products of Mg²+-ATPase are located at the outer layer of the plasma membrane of the AH 130 cells and rat liver tissue. In the isolated membrane fractions it is located at the inner layer. 5. ρ-Chloromercuribenzoate has only a slight effect on Mg²+-ATPase and Mg²+-Na+-K+-ATPase activities of the rat liver membrane, while it inhibits these enzyme activities in the AH 130 cell membrane. NaF (1 mM) and NaN3 (1 mM) inactivate ADPase of the rat liver plasma memo brane. 6. In these experimental conditions, nonenzymatic hydrolysis of ATP by lead ions is not recognized. 7. It seems most reasonable to conclude that cytochemical electron microscopic demonstration of Mg²+-ATPase after fixation with glutaraldehyde may serve as the absolute marker for the plasma membrane of ascites hepatoma and liver cells.

By means of the thin layer chromatography (TLC) a study was carried out on the decomposition of methyl parathion, ethyl parathion and sumithion when exposed to heat or ultra-violet irradiation. The results are briefly summarized as follows. 1. Parathions, when exposed to heat, form hydrolysates and such 0-analog as paraoxon as well as S-alky1 isomers. 2. When parathions are exposed to ultra-violet rays at 365 mμ and 254 mμ, the rate of decomposition is extremely slow. For example, when exposed to such rays in Petri dish for 5 hours, only a small amount of S-alkyl isomer is formed. 3. After heating parathions in a small test tube and conducting TLC, when each 0-analog and S-alkyl isomer above mentioned is confirmed, it is possible to identify a minute amount of each parathion by this method, and thus this method is feasible to apply to practical poison examination as a rapid and simple qualitative examination method.