162 • THE MICHIGAN ACADEMY OF SCIENCE. 



A PRELIMINARY REPORT UPON THE EXTRACTION OF A 

 TOXIN FROM LIVER CELLS. 



J. p. MUNSON AND FRANK E. SPENCER. 



The liver of an ox, just after the animal had been slaughtered, was 

 obtained and passed through a sausage machine, thus finely dividing the 

 structure and removing a considerable portion of the connective tissue. 

 This finely divided material was stirred up with five times its volume 

 of 1 per cent, sulphuric acid, and the mixture was heated for three hours 

 and thirty minutes in the water bath at a temperature of 81° C. Then it 

 was heated for three hours longer over the direct flame at 100° and 

 filtered. The filtrate proved to be a clear, light, amber-colored fluid, 

 which was added, drop by drop, to three volumes of 95 per cent, alcohol 

 with constant stirring. The precipitate which formed was collected 

 upon a hard filter, washed with alcohol and ether, and then dissolved in 

 a minimum amount of water. This solution was again precipitated by 

 adding it, drop b}' drop, to three times its volume of 95 per cent, alcohol. 

 The precipitate was washed with alcohol and ether, then redissolved in 

 distilled water. This process was repeated until the aqueous solution 

 failed to give a test for sulphuric acid with dimethylamidoazobenzol. 

 The substance thus obtained was dried in vacuo and ground, first in a 

 porcelain and then in an agate mortar. The cleavage product thus 

 obtained when dissolved in water and injected intraperitoneally into 

 guinea-pigs and rabbits, killed the animal when the proportion is one part 

 of the extract to 500 parts of body-weight. When used in smaller 

 quantities there is more or less marked emaciation, depending upon the 

 quantity injected, from which the animal recovers very slowly. So far 

 we have not been able to secure anv marked degree of immunitv bv be- 

 ginning with small doses and gradually increasing the quantity. 



After acute poisoning with the quantity above mentioned, post-mortem 

 examination shows the liver of the dead animal to be deeply congested. 

 The spleen is soft and mottled, with dark and pale red spots. The kid- 

 neys show no gross changes, but the adrenals are markedly congested. 

 The gastric and mesenteric vessels are greatly dilated, and there are 

 frequently found small hemorrhagic areas under the peritoneum. The 

 minimum fatal dose produces the pathological changes of an acute in- 

 toxication and kills in from twelve to thirtj-six hours. Repeated in- 

 jections of small doses produced chronic poisoning in rabbits. The 

 weight curve shows a marked fall after the first dose, but a partial im- 

 munity is soon acquired, and there is only a slight fall in weight after 

 each succeeding dose. 



We are now injecting rabbits with an emulsion of fresh liver cells of 

 a guinea-pig and Iio])e to obtain a serum from these animals, which will 

 produce cytolysis and immunity in other animals. In brief, we hope to 

 obtain a specific serum. 



J. F. MuNSON^ A. B., and Frank R. Spencer, M. D., 

 Laboratory of Hygiene, University of Michigan. 

 Ann Arbor. 



