Jpn. J. Med. Mycol. Vol. 21, , 1980 ISSN
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1 Jpn. J. Med. Mycol. Vol. 21, , 1980 ISSN
2 Fig. 1. Separation and purification procedures of cellular mannans from C. albicans and S. cereaisiae
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4 Jpn. J. Med. Mycol. Vol. 21 (No. 4),
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6 Jpn. J. Med. Mycol, Vol. 21 (No. 4), Table 3. Limulus amoebocyte lysate gelation with the toxin (Fr II-2) in comparison with the C. albicans mannan and E. coli lipopolysaccharide Fig. 4. Immunodiffusion between mannan and antiserum to C. albicans. Mannan was dissolved in phisiological saline at a concentration of 1mg/ ml. Central well: anti-c. albicans rabbit serum. A: C. albicans mannan CM-1, B: C. albicans mannan CM-2, C: S. cerevisiae mannan SM-1, D: S. cerevisiae mnnan SM-2. Table 2. Hydrolysis of C. albicans and S. cerevisiae mannans by Oerskovia sp. mannanase Reducing sugar was determined by Somogyi- Nelson method.
7 Table 7. Detection of C. albicans mannan in blood or saline by Limulus amoebocyte lysate test. Table 5. Effect of mannanase on the gelation of Limulus amoebocyte lysate by S, cerevisiae mannan a Normal rabbit blood was used. +: LAL test positive, -: LAL test negative. Table 6. Gelation of Limulus amoebocyte lysate by C, albicans formol-killed cells.
8 Jpn. J. Med. Mycol, Vol. 21 (No. 4), Table 8. Effect of toxin (Fr II-2) and C. albicans mannan on clotting time after recalcification. a Mean + standard error. Incubation temperature: 37C Table 9. Relationships between of dose of C. albicans formol-killed cells or celite and clotting time after recalcification. Assay was done after 30 minutes incubation at 37C. a Mean +standard error. Table 10. Effect of the toxin (Fr II-2) and the mannan from C. albicans on rabbit platelet aggregation in comparison with a LPS from E. coli - to ++ indicates the degree of platelet aggre- gation. Incucation temperature: 37C
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10 Jpn. J. Med. Mycol. Vol. 21 (No. 4), ) Cooperstock, MS., Tucker, R. P. and Baublis, J. V.: Possible pathogenic role of endotoxin in reye's syndrome. Lancet, June 7: , ) Cutler, J. E., Friedman, L. and Milner, K. C.: Biological and chemical characterization of toxic substances from Candida albicans. Infect. Immun., 6: , ) Elm, R. F. and Wolff, S. M.: Nonspecificity of the Limulus amoebocyte lysate test: Positive reaction with polynucleotides and proteins. J. Infect. Dis., 128: , ) Holder, I. H. and Nathan, P.: Effect in mice of injection of viable Candida albicans and a cell- extract on circulating platelats. Infect. free Immun., 7: , ) Horowitz, HI., Des Prez, R. M. and Hock, E. W.: Effects of bacteriall endotoxin on rabbit platelets. II. Enhancement of platelet factor 3 activity in vitro. J. Exp. Med., 116: , ) Isenberg, H. D., Allerhand, J. and Berkman, J. I.: An endotoxin-like fraction extracted from the cells of Candida albicans. Nature, 197: , ) Iwata, K.: Fungal toxins and their role in the etiopathology of fungal infections. In: Recent Advances in Medical and Verterinary Mycology (Iwata, K. ed.). pp , Univer- of Tokyo Press, Tokyo, sity 13) Jorgensen, J. H., Carvajal, H. F., Chipps, B. E. and Smith, R. F.: Rapid detection of gramnegative bacteria by use of the Limulus endo- toxin assay. Appl. Microbiol., 26: 38-42, ) Ballou, C. E.: Some aspects of the structure, 15) Kind, L. S., Kaushal, P. K. and Drury, P.: immunochemistry, and genetic control of yeast Fatal anaphylaxis-like reaction induced by mannan. Adv. Enzymol., 40: , yeast mannans in nonsensitized mice. Infect. 2) Bang, F. B.: A bacterial disease of Limulus Immun., 5: , polyphemus. Johns Hopkins Hosp., 98: ) Kobayashi, G. S. and Friedman, L.: Characterization 351, ) Cooper, J. F., Hochstein, H. D. and Seligman, of the pyrogenicity of Candida albicans, Saccharomyces cerevisiae and Cryptococcus neoformans. E. B.: The Limulus test for endotoxin (pyrogen) J. Bact., 88: , in radiopharmaceuticals and biologicals. Bull. Parent. Drug. Assoc., 26: , 1972.
11 274, ) Sakaguchi, O., Suzuki, S., Suzuki, M. and 18) Levin, J., Poore, T. E., Zauber, N. P, and Sunayama, H.: Biochemical and immunochemical studies of fungi. VIII. Immuno- Oser, R. S.: Detection of endotoxin in the blood of patients with sepsis due to gramnegative bacteria. N. Engl. J. Med., 283: and Saccharcmyces cereaisiae. Japan. J. Microchemical studies of mannan of Candida albicans , biol., 11: , ) Louria, D. B.: Experimental infections with 29) Salbin, S. B.: Endotoxin in pathogenic fungi. fungi and yeasts. Contr. Microbiol. Immunol., J. Immunol., 69: 88-99, : 31-47, ) Suzuki, M., Mikami, T., Matsumoto, T. and 21) Niwa, M.: The gelation reaction of horseshoe Suzuki, S.: Gelation of Limulus lysate by crab amoebocyte and endotoxin : The basic synthetic dextran derivatives. Microbial. Immunol., 21: , background and applications. Japan. J. Bactericl., 30: , ) Tripodi, D, and Nowotony, A.: Relation of 23) Niwa, M. and Waguri, O.: Biochemistry of structure of active sites in endotoxic lipopolysaccharide of Serratia marcescens. Ann. N. Y. horseshoe crab amoebocyte. 47: 1-13, Acad. Sci., 133: , ) Weiner, M. H. and Yount, W. J.: Mannan antigenenzia in the diagnosis of invasive Candida infections. J. Clin. Invest., 58: , ) Peat, S., Whelan, W. J. and Edwards, T. E.: Polysaccharides of baker's yeast. part IV. Mannan. J. Chem. Soc., Part I: 29-34, ) Ray, T. L., Hanson, A. B. S., Ray, L. F. and Wuepper, K. D.: Purification of a mannan from Candida albicans which activates serum complement. J. Invest. Dermatol., 73: 269-
12 Jpn. J. Med. Mycol. Vol. 21 (No. 4), Studies on Glycoprotein Toxins Produced by a Candida albicans Strain II. Limulus Amoebocyte Lysate-Gelling Activity of One of the GP Toxins and the Cellular Mannan Isolated from the Same Strain Yoshimasa Yamamoto and Kazuo Iwata Department of Microbiology, Faculity of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113, Japan One of the C, albicans GP toxins (Fr II-2) was demonstrated to possess a Limulus amoebocyte lysate (LAL)-gelling activity. The mannan moiety of the toxin was considered to contribute in exhibiting the activity. Purified mannans from C, albicans and Saccharomyces cerevisiae also showed a LALgelling activity; the activity of the latter was inactivated by treatment with Oerskovia mannanase. The both mannans manifested a lethal activity for mice when upon i.v, injected, being lower than the GP toxin. The GP toxin showed a pyrogenic activity, while C, albicans mannan did not. The GP toxin, mannan and C, albicans formol-killed cells were found to shorten plasma coagulation time. These results suggest that C, albicans GP toxin are classified into a category of fungal endotoxins.
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