188 Mr. J. A. Craw. On the Physical [Mar. 22, 
Table IV.—Diffusion of Lysin (0°5 Per Cent.) with Antilysin after attaining 
Equilibrium, through Gelatine Columns, showing Hemolysis, in Milli- 

metres. 
Noi. Nor 22 iiNos) i Nios 4: 4) No. bs |) a NiontGr 
oe. of porie 5 3-73 255 1°25 0 625 0°317 Saline 
fT bt yal ae net aN ae controls. 
PAD OIYS eres ae Ro oscinse tr. tr. | tr. tr. 1 °*4 2°0 0 
OMB Rare comes 1°0 1:0 1:0 1:0 15 3°5 0 

no proportionality had been found to exist between mass of lysin and effect. 
It is remarkable that although in 70 hours this effect was evident, yet in 
less than 40 hours the corresponding free lysin had a greater hemolytic effect. 
This seems most easily explained on Ehrlich’s view that lysins are complex and 
that antilysin will neutralise the most active constituents first. The same effects 
were obtained from mixtures of lysin and antilysin in the same relative 
proportions, but present in 4, 4, and th of the concentration obtaining in the 
above experiment. Controls with saline showed no effect. The partially 
neutralised mixture containing 0°5 per cent. lysin and 2°5 per cent. antilysin 
No. 3, gave the same effect as the neutral mixture and likewise a mixture of 
0-5 per cent. lysin and 1:25 per cent. antilysin, No. 4, but when the antilysin 
concentration was reduced to 0°625 per cent. (No. 5), the effect indicated 
a concentration equal to 0°125 per cent. free lysin. With 0°3175 per cent. anti- 
lysin (No. 6), and less the effect indicated 0°5 per cent. lysin free. 
These results indicate that the addition of antilysin up to 1/10th the amount 
required to entirely prevent hemolysis at 37° in the standard time (neutral 
mixture) does not appreciably neutralise the lysin. That the combination 
of lysin with antilysin is easily reversible at 18° C., does not seem 
probable, for on the addition of double the amount of antilysin the free lysin 
decreases to less than 1/4th of the total amount added. This then 
apparently conforms well with Ehrlich’s view that no neutralisation of toxin 
takes place when small quantities of antitoxin are added. 
On the Nature of the Hquilibria: Fractional Addition of Lysin to Antilysin. 
Danysz (1902) found that when a certain quantity of diphtheria toxin 
was added in fractions to antitoxin, less toxin was neutralised than in the 
case in which the whole quantity was added in one portion. This result 
has been confirmed by von Dungern (1904). Dr. C. Todd first drew my 
