X. EFFECTS OF DEFICIENCY 557 



is a normal constituent of many urines. No relation between urorosein 

 excretion and nicotinic acid malnutrition could be observed. Furthermore, 

 results of the B.E.S. test could not be correlated with the amount of 

 urinary porphyrin, and the excretion of porphyrin had no constant relation 

 to nicotinic acid deficiency. A red pigment which was observed in urine 

 from patients with pellagra and from dogs with blacktongue was identified 

 as a mixture of several related pigments similar to but not identical with 

 synthetic indirubin. These pigments were found in normal animals as well 

 as in those with nicotinic acid deficiency. Thus, it seems clear that these 

 urinary pigments have no special relationship to nicotinic acid deficiency. 



b. Urinary Fluorescent Substances 



In 1940 Najjar and Wood^^ noted a substance in urine which gave a 

 bluish fluorescence with ultraviolet light, the concentration of which seemed 

 to be increased when nicotinic acid was administered. Shortly thereafter 

 Najjar et al.^^- ^^ noted that this substance (Fo) seemed to be absent in the 

 urine of patients with pellagra and of dogs Avith blacktongue. They also 

 noted another fluorescent substance (Fi) in the urine of patients with 

 pellagra and of dogs with blacktongue which they did not find in normal 

 urine. These investigators suggested that measurement of Fi and F2 in 

 urine would be a useful diagnostic tool in pellagra. Later, Field and asso- 

 ciates,^^ Perlzweig et al.^'^ Raoul,^^ and Ellinger and Coulson^* noted similar 

 fluorescent substances in urine. The latter workers found that N^-methyl- 

 nicotinamide (nicotinamide methochloride) gave a reaction similar to F2 

 in urine. In 1943 Huff and Perlzweig'^" isolated F2 from urine and identified 

 it as N^-methylnicotinamide. Considerable controversy developed as to 

 whether the substance they isolated was identical with F2.''^"''^ In any event, 

 it is now well established that N^-methylnicotinamide is one of the principal 



«3 V. A. Najjar and R. W. Wood, Proc. Soc. Exptl. Biol. Med. 44, 386 (1940). 



6^ V. A. Najjar and L. E. Holt, Jr., Science 93, 20 (1941). 



«5V. A. Najjar, H. J. Stein, L. E. Holt, Jr., and C. A. Kabler, J. Clin. Invest. 21, 



263 (1942). 

 «6H. Field, Jr., D. Melnick, W. D. Robinson, and C. F. Wilkinson, Jr., /. Clin. 



Invest. 20, 379 (1941). 

 "W. A. Perlzweig, H. P. Sarett, and L. H. Margolis, /. Am. Med. Assoc. 118, 28 



(1942). 



68 Y. Raoul, Bull. soc. chim. hiol. 25, 266, 271, 279 (1943). 



69 P. Ellinger and R. A. Coulson, Nature 152, ,383 (1943). 



" J. W. Huff and W. A. Perlzweig, J. Biol. Chcin. 150, 395 (1943). 

 '1 J. W. Huff and W. A. Perlzweig, Science 97, 538 (1943) ; 100, 28 (1944). 

 72 J. W. Huff, J. Biol. Chem. 167, 151 (1947). 



" V. A. Najjar and V. White, Science 99, 284 (1944); 100, 247 (1944). 

 '< V. A. Najjar, V. White, and D. B. M. Scott, Bull. Johns Hopkins Hosp. 74, 378 

 (1944). 



