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tion of endotoxin and surfactants. I. Physical and biological properties of endotoxin treated with sodium desoxycholate. J. Bacteriol. 92:1493-1509. 1966.-Endotoxins from three species of gram-negative bacteria were shown to be dissociated by the bile salt sodium deoxycholate (NaD) into nontoxic subunits with molecular weights of about 20,000. When the bile salt was removed by dialysis, the subunits reaggregated in an orderly manner to form a relatively uniform population of biologically active endotoxin particles with average molecular weights of 500,000 to 1,000,000. If a small amount of human plasma was added to the dissociated endotoxin before removal of the NaD, reassociation apparently did not occur and the preparation remained nonpyrogenic. However, the plasma protein could subsequently be removed from the endotoxin subunits, and reaggregation to the toxic form would then occur. The studies on the physical nature of endotoxin performed with biophysical solution techniques were supplemented and confirmed by direct examination of the endotoxin polymers by electron microscopy. The results of these studies were consonant with the theory that the biologically active endotoxic elements are composed of micellar aggregates of linear lipopolysaccharide subunits.

Gerhardt, Philipp (The University of Michigan, Ann Arbor), and Edgar Ribi . Ultrastructure of the exosporium enveloping spores of Bacillus cereus . J. Bacteriol. 88: 1774–1789. 1964.—Structural details in the outer envelope of spores, such as those of Bacillus cereus and B. anthracis , were studied by electron microscopy and by X-ray diffraction analysis. Procedures were developed for isolating homogeneous fragments of the membrane with minimal damage to or germination of the spore proper. Exosporium of B. cereus appeared to embody two main layers. An outer layer was made up of a nap of hairlike projections, irregularly distributed and about 250 A deep; these arose from an intermediate covering, about 60 A in depth and similarly lead-stainable. An inner basal layer had a hexagonally perforate surface pattern of holes, averaging 76 A from center to center, and was made up of four lamellae, which could fragment into crystal-like elements. The intact basal membrane was about 190 A thick and the thinnest elements, 45 A. Microscopic observations of a crystal-like nature of the exosporium basal membrane were confirmed by X-ray diffraction analysis; the pattern of reflection lines in powder diagrams of exosporium fragments or paracrystals, or intact spores, corresponded to a hexagonal, close-packed crystal structure. The unit cell was calculated to have dimensions of 7.6 A along the a axis and 11.9 A along the c axis of the space lattice.

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