INTRACELLULAR CHANGES AFTER PHENYLHYDRAZINE 525 



was erroneously considered as hemoglobin by Warburg and col- 

 laborators ("2944), who were forced to assume that it was a peculiar 

 kind of hem/globin, since the erythrocytes remained brown on reduc- 

 tion. The significance of their finding that denatured globin was 

 also present is discussed below. Lemberg and Legge (1704) showed, 

 however, that after an injection of 50 mg. per kg. phenylhydrazine 

 into a rabbit, choleglobin could be detected in greatly increased 

 amounts and biliverdin could be isolated from the cells. The bili- 

 verdin concentration reached a maximum value of 11.7 mg. per 100 

 ml. of cells after 4 hours, and reached normal levels between the 

 second and sixth day after injection. The real biliverdin concentra- 

 tion in the cells was probably about twice as high since the recovery 

 is not quantitative. During the experiment the hemoglobin level 

 fell by about one third. Cruz (514) has observed a considerable 

 decrease in the mean corpuscular hemoglobin in the dog one day 

 after injection of acetylphenylhydrazine. The choleglobin-containing 

 erythrocytes are rapidly removed from circulation (1527). 



Phenylhydrazine and aromatic amino and nitro compounds have 

 long been known to form inclusion bodies in the erythrocyte, known 

 as Heinz bodies after their discoverer (1230). They were associated 

 with the breakdown of the hemoglobin which occurs under these con- 

 ditions and, more specifically, with hemzglobin formation (1967), 

 "verdohemochromogen" formation (1439), and with Warburg's dis- 

 covery of the presence of intracorpuscular denatured globin. Heubner 

 (1255,1256), Jung (1445), and Kiese and Seipelt (1529) conclude that 

 they are due to the denaturation of stroma protein. It appears 

 more likely, however, that they contain denatured globin. This is 

 well supported by Horecker's observation (1344) that the formation 

 of Heinz bodies in dog erythrocytes after trinitrotoluene adminis- 

 tration coincides with the appearance of a precipitate of denatured 

 globin in hemolyzed blood; by dialysis this is reconvertible into 

 native globin. Denatured globin cholehemochrome has also been 

 found (1529). The claim of Heubner that there is no correlation 

 between formation of choleglobin ("verdoglobin") and Heinz bodies 

 is based on the erroneous equation of sulfhemoglobin and choleglobin 

 (c/. Section 5.4.). 



The appearance of denatured globin within the cell is probably to 

 be associated with the observations made by Lemberg and Legge 

 (1704) that more biliverdin was found in rabbit cells after phenyl- 

 hydrazine than could be accounted for by their choleglobin content. 



