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Technique for the purification of the hyphae of Filobasidiella neoformans

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A technique for the purification of the hyphae ofFilobasidiella neoformans is described. Cultural conditions and strains for maximal hyphal production were determined. Blastospores were separated from hyphae by sonification of mycelial suspensions. Following this, the initial density ranges of all cell types were determined by isopycnic centrifugation, using Renografin-60 as a supporting medium. Based on these data, rate centrifugation with different density ranges was used to obtain cell separation. The final percentage of non-hyphal cells to total population was 0.007%. These were determined to be still viable at the end of separation.

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  1. 1.

    Bowman, P. I. & Ahearn, D. J., 1977. Ecology of Cryptococcus neoformans in Georgia. Symposium: the black and white yeasts. Pan. Am. Health Org. Pub. No. 25.

  2. 2.

    Brown, H. J., Cook, K. M., Ney, T. T. & Hatch, T., 1950. Influence of particle size upon the retention of particulate matter in the human lung. Am. J. Pub. Health. 40: 450–458.

  3. 3.

    Davidson, E. A. & Rosett, T. Ultrasonic disruption of bacteria and other small microorganisms. Bronwell Sonifier Instruction Manual.

  4. 4.

    Dykstra, M. A., Friedman, L. & Murphy, J., 1977. Size of Cryptococcus neoformans: control and relationship to virulence. Infect. and Immun. 16: 129–135.

  5. 5.

    Edwards, M. R., Gordon, A. R., Laper, E. W. & Ghiorse, W. C., 1967. Micromorphology of Cryptococcus neoformans. J. Bact. 94: 766–777.

  6. 6.

    Erke, K. H., 1976. Light microscopy of basidia, basidiospores, and nuclei in spores and hyphae of Filobasidiella neoformans (Cryptococcus neoformans). J. Bact. 128: 445–455.

  7. 7.

    Farhi, F., Bulmer, G. J. & Tacker, J. R., 1970. Cryptococcus neoformans: the not-so-encapsulated yeast. Infect. & Immun. 1: 526–531.

  8. 8.

    Fisher, W. D., Cline, G. B. & Anderson, N. G., 1964. Density gradient centrifugation in angle-head rotors. Analyt. Biochem. 9: 477–489.

  9. 9.

    Forman, L., 1978. The quantitation of hemoglobin transition during normal and thyroid induced metamorphosis in the bullfrog Rana catesbiana. Ph.D. Dissertation. University of Kentucky.

  10. 10.

    Grdina, D. J., Milas, L., Hewitt, R. R. & Withers, H. R., 1973. Buoyant density separation of hyman blood cells in Renografin gradients. Exp. Cell Res. 81: 250–254.

  11. 11.

    Hatch, T. T., 1961. Distribution and deposition of inhaled particles in the respiratory tract. Bact. Rev. 25: 237–240.

  12. 12.

    Hass, F. J., Lee, P. S. & Lourence, R. V., 1978. Regional lung deposition of inhaled particles. J. Clin. Res. 26: 687a.

  13. 13.

    Kwon-Chung, K. J., 1975. A new genus, Filobasidiella, the perfect state of Cryptococcus neoformans. Mycologia 67: 1197–1200.

  14. 14.

    Kwon-Chung, K. J., 1976a. Morphogenesis of Filobasidiella neoformans, the sexual state of Cryptococcus neoformans. Mycologia 68: 821–833.

  15. 15.

    Kwon-Chung, K. J., 1976b. A new species of Filobasidiella, the sexual state of Cryptococcus neoformans, B and C serotypes. Mycologia 68: 942–946.

  16. 16.

    Kwon-Chung, K. J., 1978. Heterothallism vs. self-fertile isolates of Filobasidiella neoformans (Cryptococcus neoformans). Proc. IV Int. Conf. Mycoses, Brasilia, Brazil. PAHO Sci. Publ. 356-204-213.

  17. 17.

    Kwon-Chung, K. J. & Bennett, J. E., 1978. Distribution ofα anda mating types of Cryptococcus neoformans among natural and clinical isolates. Am. J. Epidem. 108: 337–340.

  18. 18.

    Larsh, H. W., Scholes, V. E., Hinton, A. & Silberg, S., 1958. Minimal infectious inoculum of Histoplasma capsulatum for mouse and chick embryo. Proc. Soc. Exp. Biol. Med. 98: 570–573.

  19. 19.

    McCall, J. S. & Potter, B. J., 1973. Ultracentrifugation. Williams and Wilkins, Baltimore.

  20. 20.

    Mrak, E. M., Phaff, H. J. & Douglas, H. C., 1942. A sporulation stock medium for yeast and other fungi. Science 96: 432.

  21. 21.

    Neilson, J. B., Fromtling, R. D. & Bulmer, G. S., 1977. Cryptococcus neoformans: size range of infectious particles from aerosolized soil. Infect. & Immun. 17: 634–638.

  22. 22.

    Powell, K. E., Dahl, B. A., Weeks, R. J. & Tosh, H. E., 1972. Airborne Cryptococcus neoformans: particles from pigeon excreta compatible with alveolar deposition. J. Infect. Dis. 125: 412–415.

  23. 23.

    Sena, E. P., Radin, D. N., Welch, J. & Fogel, S., 1975. Synchronous mating in yeasts. Chapter 4. Methods in Cell Biology 11: 71–88. Academic Press, N.Y.

  24. 24.

    Shadomy, H. J. & Lurie, H. L, 1971. Histopathological observations in experimental cryptococcosis caused by a hypha-producing strain of Cryptococcus neoformans (Coward strain) in mice. Sabouraudia 9: 6–9.

  25. 25.

    Shadomy, H. J. & Utz, J. P., 1966. Preliminary studies on a hypha-forming mutant of Cryptococcus neoformans. Mycologia 58: 383–390.

  26. 26.

    Smith, C. D. & Furculow, M. L., 1964. Evidence of the presence in yeast extract of substances which stimulate the growth of Histoplasma capsulatum and Blastomyces dermatitidis similar to that found in starling (Sturnis vulvaris) manure. Mycopath. Mycol. Appl. 22: 99–105.

  27. 27.

    Utz, J. P., Shadomy, H. J. & Lurie, H. I., 1969. Cryptococcus neoformans and cryptococcosis. Trans. Amer. Clin. Climatol. Assn. 81: 220–226.

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Zimmer, B.L., Hempel, H.O. & Goodman, N.L. Technique for the purification of the hyphae of Filobasidiella neoformans. Mycopathologia 73, 171–176 (1981). https://doi.org/10.1007/BF00575088

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  • Purification
  • Centrifugation
  • Cultural Condition
  • Total Population
  • Initial Density