Abstract
Several investigators have found that hemorrhage could be produced in tumors by the administration of bacterial preparations and that the tumors subsequently receded in a significant proportion of cases. Confirmation of these findings has been reported. 1 , 2 However, the results obtained were not satisfactory inasmuch as small doses were often ineffective and large doses were toxic. The separation of the hemorrhage-producing agent from toxic and inert contaminants was therefore undertaken.
Little progress has hitherto been reported in this direction. The most recent relevant contribution is that of Apitz 3 who attempted to purify the “Shwartzman-active”substances present in B. typhosus and B. paratyphosus B preparations. Partial purification was effected, but Apitz found that the Shwartzman-active material so obtained was highly unstable; this instability halted further purification. It should be pointed out however that, in this study, Apitz was concerned solely with the Shwartzman phenomenon and reported no observations on the effect of this material on tumors.
The agent in B. coli filtrate responsible for hemorrhage production in mouse tumors was found to be stable in most instances, in this laboratory, even after a high degree of purity had been attained. The first steps in the separation of the active fraction were based upon the method devised by Felton, Kauffmann and Stahl 4 for the precipitation of the soluble specific polysaccharide from pneumococcus broth cultures.
The method used was, briefly, as follows:
The organisms were removed from week-old nutrient broth by centrifugation and by Berkefeld filtration. Phosphate was added to the filtrate; then the pH was brought to 9.5 by addition of lime water. The active material was carried down with the basic calcium phosphate precipitate, but a single precipitation did not always suffice to bring it down completely.
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