654 



SCIENCE 



[N. S. Vol. XXVII. No. 695 



journal. It was further shown by Dr. 

 Abel and myself that this Amanita-hemo- 

 lysin is not a tox-albumin as Kobert had 

 stated, since all proteid can be removed 

 from it, by the use of freshly prepared 

 metaphosphorie acid, and by uranyl 

 acetate, without any appreciable impair- 

 ment of its hemolytic activity. Our hemo- 

 lytic solution thus treated no longer re- 

 sponds to any of the well-recognized tests 

 for either native or derived proteids. 

 Although this hemolysin has by no means 

 been obtained in a condition of chemical 

 purity, it must, for the present at least, be 

 classified as a glucoside because of the fol- 

 lowing reactions which our purest sub- 

 stance gives. 



1. It reduces Fehling's solution and 

 ammoniacal silver solution onlj^ very 

 slightly without previous hydrolysis with 

 acids, and very poAverfuUy after such 

 hydrolysis. 



2. It does not ferment with brewers' 

 yeast either before or after hydrolysis. 



3. It gives characteristic tests for pen- 

 toses with (a) a-napthol and sulphuric 

 acid, (&) phloroglucinol and hydrochloric 

 acid, (c) orcinol, hydrochloric acid, and 

 ferric chloride. It also decolorizes an 

 alkaline solution of potassium permanga- 

 nate at room temperature, and after 

 hydrolysis gives a yellow color with sodium 

 hydrate. 



The alcohol-soluble Amanita-toxin, which 

 proba;bly is more important in cases of 

 poisoning in man because of its resistance 

 to the action of heat and acids than the 

 Amanita-}iem.olysm has been shown by Dr. 

 Schlesinger and myself to be either an 

 indol or pyrrol derivative or an aromatic 

 phenol so combined with an amine gi'oup 

 that it readily forms an indol or pyrrol 

 ring on fusion. This substance can be ob- 



• Schlesinger and Ford, The Journal of Biolog- 

 ical Chemistry, Vol. III., No. 4, September, 1907. 



tained free from both the glucosides and 

 the native proteids present in the plants. 



Since publishing these various observa- 

 tions a number of experiments have been 

 completed which confirm our earlier con- 

 clusions and throw light upon some of our 

 difficulties, and it is desirable at the present 

 time to take up these further experiments 

 in brief detail. In the first place. Dr. 

 Kinyoun while at the Mulf ord Laboratories 

 at Glenolden, Pa., was good enough to im- 

 munize a horse for me with aqueous ex- 

 tracts of Amanita pJialloides and found 

 that its serum contained anti-bodies for the 

 poisons of this fungus of such a strength 

 that one fourth of a cubic centimeter would 

 neutralize the poisonous dose for a 500- 

 gram guinea-pig. While this is of hardly 

 more than theoretical value, in studying 

 this serum during the past year we have 

 found that it contains a strong and per- 

 manent anti-hemolysin operative in a dilu- 

 tion of 1/1,000, using as an index that 

 quantity of hemolysin which will dissolve 

 1 cubic centimeter of a 5-per-cent. suspen- 

 sion of blood corpuscles, and this anti- 

 hemolysin is still present, even though the 

 serum is nearly a year old. 



Again, it has been shown that both the 

 J.mam'ia-hemolysin and the Amamta-toxin 

 are poisonous to small animals, the lesions 

 produced by the latter substance being 

 similar to those seen in fatal cases of 

 poisoning in man. The Amanita-hemolysm 

 apparently owes its toxicity entirely to its 

 blood-making properties, the pure toxin 

 acting as a cellular poison, producing both 

 the hemorrhages and the fatty degenera- 

 tion. 



The Antanita-hemolysin, moreover, tends 

 to lose its activity on heating to 65° C. for 

 one half hour and may play but a secondary 

 role in fatal cases in man, the toxin 

 possibly being the more important prin- 

 ciple. Various animals have been immu- 



