I9I2] Edgar G. Miller, Jr. and William J. Gies 479 



of treatment was reversed — 24 hours in water, six minutes In boil- 

 ing alcohol and one hour in the latter as it cooled. 



At the end of the " edema period " the triturated tissue in each 

 case was placed in 200 c.c. of water and aqueous extraction was 

 continued for 24 hours. The corresponding extracts were com- 

 bined before analysis was begun. 



Foiirth series: With kidneys and heart. The results of the 

 third series of tests did not bear out the assumption on which the 

 special treatment was based. To our surprise the edematous tis- 

 sues yiclded smaller quantities of " non-coagulable nitrogen " than 

 the Controls. At this point our table showed three plus balances 

 and three minus balances.'^ It was then assumed that the produc- 

 tion of non-coagulable protein would increase greatly during the 

 period of shrinkage and softening that follows the attainment of 

 the maximum degree of bloating in such experiments. In the 

 fourth series of tests we accordingly duplicated the conditions of 

 the third, except that the "edema period" was 72 hours instead of 

 24 hours, and the water surrounding the tissues was renewed at 

 the end of each of the three days.^ All the corresponding extracts 

 were combined before the analyses were begun. 



The result of the test on kidney is striking. The edematous 

 kidney gained a fairly large proportion of "non-coagulable nitro- 

 gen." This particular mass of edematous tissue was the only one 

 (IV-V) that lost considerable weight after a period of 24 hours. 

 Possibly this loss of weight was due in part to enzymic formation, 



' Every endeavor was made to prevent analytic error or irregularities in 

 duplicate processes. It is possible that the preliminary treatment of the control 

 tissue with hot alcohol in the third series was more effective in preparing the 

 tissue for complete extraction than the similar treatment of the edematous 

 tissue at the end of the experiment. The control tissue could be more finely 

 triturated because, even after the concluding alcohol treatment of the edematous 

 tissue, the latter was softer, apparently more hydrous, and probably more reten- 

 tive of extractable substances. We are fully aware, also, of the difficulties 

 besetting quantitative removal of such substances as proteoses and peptones 

 from tissues. For these reasons we do not lay much stress on the differences 

 for " non-coagulable nitrogen" in any of the tests of the first three series of 

 experiments. 



* This course of treatment prevented cumulative increase of an edema- 

 favoring acidity, but it also effected the removal of such inhibitive factors as 

 salins. 



