We plan to produce amebae of the different strains in quantity to be used for the preparation of high titered chromatographed NACM. The NACM will be used basically in two ways, in the purification and characterization studies, and in the biological studies. The usefullness of isoelectrofocusing for further purification will be explored more fully, as will the techniques of ion-exchange chromatography and electrophoresis. As the methods are evaluated and incorporated into the purification scheme, the products will be subjected to further physical characterization by spectrophotometric and enzymatic techniques. The biological activity will be monitored by tissue culture assays. For the cytological studies of the effects of NACM in cultured cells, we hope to obtain antibody by the inoculation of hamsters with the semi-purified NACM. A comparison of the effects of the amebae of the different strains with the NACM prepared from the same strains on mice will be continued. BIBLIOGRAPHIC REFERENCES: Dunnebacke, T.H. and Schuster, F.L., The nature of a cytopathogenic material present in amebae of the genus Naegleria. Amer. J. Trop Med. and Hyg. 26: 412-421, 1977. Dunnebacke, T.H. and Schuster, F.L., Cytopathogenic material from the amebae of the genus Naegleria. Microbiology, 1977.