6 SECTION I. HYGIENIC MICKOBIOLOGY AND PARASITOLOGY. 



II. HISTOLOGICAL STUDIES OF THE TISSUES IN IMMUNIZED ANIMALS WITH 

 A COMPARISON OF CHEMICAL AND MICROCHEMICAL TESTS FOR GLYCO- 

 GEN. 40 



The histopathological studies of Gay and Southard 41 in serum 

 anaphylaxis in the guinea pig have seemed an interesting contribu- 

 tion to the relation of structure to function. These authors found 

 that anaphylactic intoxication is accompanied by definite lesions in 

 the nature of hemorrhages and more particularly by fatty changes 

 in parenchyma and in the endothelium. These endothelial fat 

 changes could be produced in a few minutes following intravenous 

 injection and obviously bear direct relation to the cyclonic symptoms 

 of the syndrome. It occurred to us that similar evidence of func- 

 tional cellular activity might be histologically demonstrable in the 

 cells that are engaged in antibody formation. For this purpose 

 rabbits were highly immunized by repeated intravenous injection of 

 washed guinea-pig corpuscles and following a rest of two or more 

 weeks were given a critical reinjection of 1 cubic centimeter of 

 washed guinea-pig blood and at subsequent intervals bled and the 

 tissues fixed in various ways and stained by many methods. The 

 critical reinjection was aimed, obviously, to obtain the antibody- 

 producing cells in a condition of highest activity. 



In the first experiment a series of immunized rabbits received 

 each the critical reinjection of antigen and were then bled at 1, 4. 

 and 24 hours and 4 and 6 days subsequently. A careful histological 

 study of tissues from this series showed in the 24-hour animal a very 

 marked increase of glycogen in the liver (Best's carmine stain, alco- 

 hol fixation). The animals bled before and after this period gave 

 a regular increase and decrease of glycogen to the 24-hour maximum. 

 A more careful control of this experiment has involved us in techni- 

 cal difficulties that seemed too time consuming and led, moreover, to 

 far less encouraging results than the first experiment. The question 

 of the effect of starvation on the amount of glycogen alone seemed 

 unsurmountable ; it was found, for example, that rabbits that have 

 not been fed for two days have stomachs stuffed with food. We are 

 still in doubt as to the significance of our first find of increased 

 glycogen. An attempt to correlate the microchemical reaction of 

 glycogen with a careful chemical analysis of total glycogen in the 

 liver has, however, led to results of importance. So far as we are 

 aware, there has been no systematic comparison between the chemical 

 analysis of an organ and microchemical staining reactions in a sam- 

 ple of it. Dr. Rusk has studied the amount and distribution of 

 glycogen in sections of 22 rabbit livers stained by the Ehrlich iodin 

 method and the Best carmine method, with a chemical analysis of a 

 greater (weighed) portion of the same livers, following Pfluger's 



