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the kindness of G. Wallmark); normal human and rabbit sera; bacterial (S. aureus, L. monocytogenes, and B. subtilis) antisera from rabbits; and sera from subjects immunized with staphylococcal antigen (courtesy of T. R. Neblett). Details of the basic and protease hemagglutination tests have been described previously (Neter et al. Proe. Soc. Exptl. Biol. Med. 101:484, 1959). Briefly, washed human erythrocytes (2.5%, blood group 0) were mixed with antigen in various concentrations in phosphate buffer (Difco) or in pancreatic protease (62 ,g/ml). After incubation at 37 C for 30 min, the erythrocytes treated with antigen-buffer and antigen-protease were washed and added to serum and gamma globulin in serial dilutions. The mixtures, after 30 min at 37 C, were centrifuged (3 min at 1,200 X g), and the resulting hemagglutination was read grossly. The effect of pH and heat on heterogenetic (staphylococcal) antigen, as determined by hemagglutination, is shown in Table 1. With antigen heated at alkaline pH, there was a decrease in the titer of hemagglutinins in normal and immune gamma globulin and in normal serum, when compared to acid-heated and unheated antigen preparations. In contrast, when bacterial antisera were employed, all antigen preparations were highly active, independent of heat and pH treatment, even with minimal amounts of antigen. Study of L. monocytogenes and B. subtilis antigens (not shown in table) yielded similar results. The possibility

ROLE OF METALS IN MANOMETRIC STUDIES WITH CEPHALOSPORIUM LONGISPORUM