35 



but in an aqueous solution of the (NH 4 ) 2 SO 4 -saturation pre- 

 cipitate from the urine, that if a larger amount of substance 

 had been present, the heat-precipitation test would have been 

 obtained in the urine itself. 



D. LEUCOCYTIC PROTEOLYSIS OF OSSEOALBUMOID. The orig- 

 inal assumption which led to this work was that the proteolytic 

 enzymes in the cells of the new bone growth, or the unrestrained 

 action of the enzymes from the bone marrow cells, might cause 

 the production of the Bence Jones protein by the transformation 

 of osseoalbumoid. It was desirable, therefore, to ascertain 

 what the direct action of the enzymes of leucocytes would 

 be on osseoalbumoid and to learn the results that would occur 

 if the substances produced by their proteolysis were injected 

 into dogs. 



The method described by Opie (Jr. Exp. Med., 1906, viii, 

 p. 410) was used to obtain the leucocytes and to separate 

 the enzymes from them in a dry, powdered state. By this 

 method Opie has shown that an enzyme designated by him 

 "lymphopro tease" is destroyed when present in an acid medium, 

 and that one which he calls "leucoprotease" remains active 

 when contained in an alkaline medium. The alkaline condi- 

 tion was favorable for our proposed experiment, as the leuco- 

 protease would, in bone marrow, normally act in an alkaline 

 medium. Opie has shown also (Jr. Exp. Med., 1905, vii, 

 p. 759) that the enzymes from bone marrow are identical 

 with those obtained from polynuclear leucocytes with fine 

 granulations, and, since these cells are formed in the bone 

 marrow, the bone marrow may be regarded as the source of 

 the enzymes which both contain. Furthermore, Opie has 

 shown that the "lympho-pro tease" is contained in the lympho- 

 cytes, and although the greater proportion of the cells in an 

 inflammatory exudate are polynuclears, there are a number 

 of lymphocytes present, and by his method of preparation, 

 the enzymotic action of the lymphocytes is eliminated. 



In the experiment described below a portion of osseoal- 

 bumoid was treated with a suspension of 2.35 grams of dry, 

 powdered leucocytes in saline solution. About 800 cc. of 

 0.25 per cent. N^COg and i cc. of toluol were added and the 



