I9I2] Edgar G. Miller, Jr. and William J. Gies 477 



the development of edema ranged f rom 24 to 72 hours. An " edema 

 period " of 24 hours was sufficient for our purposes in most of the 

 experiments. Toluene or Chloroform, or both, were used as pre- 

 servatives, when occasion required. The mixtures were gently 

 shaken occasionally during the edema periods. 



At intervals during the " edema period," or at its dose, the tissue 

 masses were suspended above the open Containers, extraneous Hquid 

 was allowed to drain back into the main volumes and the weights of 

 the bloated masses were recorded. At the end of each " edema 

 period" the corresponding edematous tissue was subjected to the 

 conditions which had been appHed to the control tissue for the pre- 

 vention of edema. This was done in order to equaHze, if possible, 

 any hydrolytic or other disturbing influences that might have been 

 due to the different manipulations. 



Promptly after the conclusion of each "edema period," the 

 tissue was minced and then finely triturated with sand in a large 

 mortar. Quantitatively comparable aqueous extracts and iiltrates 

 were prepared, under identical conditions in each series of tests. 

 The mixtures were thoroughly shaken repeatedly during the "ex- 

 traction periods." 



AHquot portions of the neutralized filtrates were carefully freed 

 from coagulable protein by the heat coagulation process.* "Non- 

 coagulable nitrogen " in the fihrates was determined by the Kjeldahl 

 method. 



The special details of each series of experiments are stated in 

 the ensuing descriptions. The analytic data are summarized in the 

 table on page 480. 



Experiments. First series: With kidneys. The control kid- 

 ney was placed in 100 c.c. of water + 100 c.c. of 95 per cent. alco- 

 hol. The second kidney was immersed in 100 c.c. of water. It 

 was assumed that the alcohol would prevent appreciable hydrolysis 

 in the control kidney. These conditions were maintained for 24 

 hours. At the end of that time, 100 c.c. of 95 per cent. alcohol 

 were added to the liquid containing the edematous kidney. The 

 extraction volumes were then equal. The new conditions prevailed 



*We added dilute acetic acid Solution, little by little while the liquid was 

 hoiling (not before), in order to effect complete coagulation. 



