47^ Enzymes as Factors in Edema [Mar. 



from the beginning of the 25th to the 72d hours, inclusive. At the 

 end of this period the alcoholic liquids were poured off and retained. 

 In each case the macerated tissue was further extracted for 48 

 hours with 500 c.c. of 5 per cent. sodium chloride Solution. The 

 alcoholic and salin extracts from each tissue were combined before 

 the analytic procedura was started. 



Second series: With kidney^, testicles and heart.^ The tissues 

 were subjected to conditions which were essentially the same as 

 those that prevailed in the first experiment, but 20 per cent. sodium 

 Chloride Solution was used instead of 95 per cent. alcohol in the pre- 

 liminary treatment and instead of 5 per cent. sodium chloride Solu- 

 tion for the final extractions. 



Third series: With kidneys and heart. The results of the fore- 

 going tests (see the data on page 480) suggested that, in all but one 

 instance, sufficient hydrolysis of protein occurred in the edematous 

 tissues to give them perceptibly larger Contents of non-coagulable 

 nitrogenous substances. We then assumed that our treatment for 

 the total inhibition of enzymic hydrolysis in the control tissues had 

 not been wholly successful in the first two series of tests, and con- 

 cluded that the observed difference between the corresponding 

 amounts of " non-coagulable nitrogen" would be greater if enzymic 

 cleavage were completely prevented in new control masses. In this 

 series of tests we accordingly aimed to destroy all enzymes in the 

 control tissue, with the aid of heat but without causing hydrolysis 

 by the heating process. 



The control masses were dropped into hoiling 95 per cent. 

 alcohoP in covered beakers on water baths. The boiling was con- 

 tinued for six minutes. After the alcoholic extract became cold (an 

 hour later), the tissue was kept in 150 c.c. of water about 24 hours. 

 For the second portion of each tissue ("edema tissue"), the order 



" In this series and in all others that included experiments on heart, the 

 organ was divided longitudinally through the auricles and ventricies into approx- 

 imately equal masses, one of which was used for control purposes, the other 

 for the edema tests. 



• We selected alcohol for this purpose because of its dehydrating influence. 

 At the temperature of boiling alcohol under the imposed conditions, the tissue 

 enzymes must have been destroyed, the coagulable proteins were coagulated and 

 little if any hydrolysis could have taken place. 



