VI. EFFECT OF THE LIPIN MATERIAL ON HEMOLYSIS 



i. RECORD OP EXPERIMENTS: Since the ratios of cholesterol to phos- 

 phatide are different for the different types of streptococci it was interest- 

 ing to investigate the effects which the lipin mixtures would have on the 

 hemolytic action of saponin.* A 10 per cent washed rabbit cell suspen- 

 sion was used in the experiments. The method used was essentially that of 

 Ranson (69). Preliminary titrations were made to obtain the dose of 

 saponin that will hemolyze I .o c.c. of cell suspension in the presence and 

 absence of cholesterol (the amount of cholesterol used approximated that 

 in the lipin mixture) . That amount of saponin which produced hemolysis 

 in the absence of cholesterol but which failed to do so in those tubes which 

 contained cholesterol was taken as the test dose. This dose was then 

 added to graded amounts of lipin material dissolved in ether. The pro- 

 tocols below show in detail the experiments performed and the results 

 obtained. 



Experiment (a): To determine the largest amount of saponin that 

 produces hemolysis in the presence of cholesterol. 



Incubation at 25 C. for 15 Minutes 



This experiment shows that in the presence of cholesterol 0.08 cc. of 

 a 0.2% saponin solution is the largest amount that does not hemolyze 

 i.o c.c. cell suspension in 8 minutes. 



Experiment (b): To determine the smallest amount of saponin that 

 produces hemolysis in the absence of cholesterol . 



Incubation at 25 C. for 15 Minutes 



Cell Suspension i.o i.o i.o i.o i.o i.o i.o i.o i.o 



Readings: 

 8 minutes 



15 minutes 



60 minutes 



12 hours 



*The saponin was obtained from Eimer and Amend, New York City. 



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