and Laboratory Methods. 1137 



4. Attempts to isolate parasitic micro-organisms from malignant tumors. 



First. In the histological study tissues were hardened in alcohols of various 

 strengths, Hermann's solution, Flemming's solution, corrosive sublimate, and 

 Zenker's fluid. Zenker's fluid gave the best results. Paraffin imbedding was 

 used. 



For staining, the methods recommended by Sanfelice and PUmmer were 

 tried at first, but as they did not give satisfactory results, the following method 

 suggested by Mallory was used : 



1. Ten per cent. aq. sol. ferric chloride, two minutes. 



2. Aq. sol. haematoxylin (1-2 per cent.), freshly made, two minutes. 



3. Wash in water. 



4. One per cent. sol. ferric chloride until blue color is removed 

 from protoplasm and nuclear stain is distinct. (Watch under 

 microscope.) 



5. Wash in water. 



6. In the following solution for two minutes. 



1 per cent. aq. sol. acid fuchsin, one part. 

 Sat. aq. sol. picric acid, two parts. 



7. Wash in water. 



8. Ninety-five per cent, alcohol. 



9. Xylol, three changes, blotting between each change. 

 10. Mount in Xylol balsam. 



This stain colors nuclei black, protoplasm a faint greenish pink, and con- 

 nective tissue a brilliant red. The peripheral and central portions of inclusions 

 stain a brilliant red, the intermediate portion a faint pink. 



The stain is simple in manipulation, constant and even in its results. 



Thirty-five carcinomata and five sarcomata were examined. In seventeen 

 cases, bodies similar to those described by Sanfelice and Plimmer were found. 

 They were found principally in cancer of the breast, in thirteen out of sixteen 

 cases. They were never found in epidermoid cancer (thirteen cases) nor in 

 sarcoma (five cases). 



Although the writer makes no definite negative statement, he apparently 

 believes that these bodies have nothing to do with the causation of cancer. 



Second. Inoculations were made from tumors which were received within two 

 hours after operation, and which were not ulcerated. Under aseptic precau- 

 tions portions of tumor were removed and placed in the peritoneal cavity of a 

 rabbit and a guinea-pig. In all, nine rabbits and three guinea-pigs were inoc- 

 lated, chiefly with tissues from cancer of the breast. All inoculations were 

 negative. 



Third. Subcutaneous and intraperitoneal inoculations and injections into ear 

 vein, liver, and anterior chamber of the eye of rabbits and of guinea-pigs, of the 

 " Saccharomyces neoformans " of Sanfehce and the micro-organism of Plim- 

 mer produced only inflammatory and proliferative changes. No tissue resem- 

 bling cancer was produced. 



Fourth. Cultures were made from thirteen cases. In three, pyogenic cocci 

 grew, the other ten remained sterile. 



