198 



SCIENCE. 



[Vol. XIX. No. 479 



TOX-ALBUMIN DIPHTHERIA.' 



In a preliminary communication published in the Hospital 

 Bulletin, No. 15,° we called attention to the histological 

 changes in the organs of animals which had died of experi- 

 mental diphlhei'ia, following the inoculation of pure cultures 

 of the bacillus diphtheriae. Since then we have extended 

 our investigations so as to include the study of the lesions 

 produced by the inoculations of the toxic products of the 

 diphtheria bacillus. This study virtually finishes the work 

 we have undertaken, and it is hoped soon to publish our re- 

 sults in detail. However, in order to make our preliminary 

 communication complete, we append this report: — 



The toxic products of the diphtheria bacillus with which 

 we have operated were obtained by filtering through a new 

 and sterilized Chamberland filter the culture of the organisms 

 in glycerine bouillon, several weeks old. The fluid so ob- 

 tained was tested by means of cover-slips and inoculations 

 on glycerine-agar, and proved to be sterile. 



Guinea pigs were used for the experimental inoculations. 

 The sterile culture fluid was introduced subcutaneously into 

 the tissues of the belly wall by means of a Koch's syringe. 

 The method pursued Will be given in connection with the 

 case of which the lesions are to be described. The guinea 

 pig received on Dec. 10, 1891, one cubic centimetre of the 

 filtrate. Not having succumbed on Dec. 14, it received two 

 cubic centimetres more. The animal died on Jan. 5, 1892, 

 the duration of life since the first inoculation having been 

 three weeks and five days, and since the last three weeks and 

 one day. 



At the autopsy the vessels of the subcutaneous tissues were 

 injected, and hemorrhage had taken place into the tissues of 

 the axillary and inguinal regions. The subcutaneous tissues 

 were moist, but there was bo actual oedema present. Neither 

 was there a visible area of localized inflammation ; no micros- 

 copical examination was made of the seat of inoculation. 

 The lymphatic glands of the axillary and inguinal regions 

 were enlarged and reddened, the cervical glands were swollen, 

 and the thyroid gland was greatly congested. 



There was a considerable excess of clear fluid in the peri- 

 toneal cavity. Both layers of the peritoneum were reddened, 

 the vessels of the visceral layer being especially injected. 

 The spleen was enlarged to double the average size. It was 

 mottled, and the white follicles were distinctly outlined 

 against the red ground. The liver was dark in color, and 

 contained much blood. On the surface a prominent yellowish- 

 white area two millimetres in diameter, surrounded by a zone 

 of hyperaemia, was observed. Smaller dot-like points of 

 the same color and general appearance were seen elsewhere 

 in the liver. The kidneys were congested and the cut sur- 

 face was cloudy. The adrenal glands appeared normal, as 

 did the mesenteric glands. 



The pleural cavity did not contain such a marked excess 

 of fluid. The pericardial sac, however, was distended with 

 clear serum. Under the epicardium were many ecohymotic 

 spots. The lungs exhibited areas of intense congestion, or 

 actual hemorrhage into the tissues. The glands of the thorax 

 were, perhaps, swollen. 



The examination of the heart muscle by means of frozen 

 sections showed it to be slightly fatty. The epithelium of 

 the tubules of the kidney was extremely granular and much 



' " The Histological Lesions Produced by the Tox-Albumln of Diphtheria," 

 by Wm. H. Welch, M.D., professor of pathology, and Simon Flexner, M.D., 

 fellow In pathology. From the Pathological Laboratory of the Johns Hopkins 

 University and Hospital. Bulletin of the Johns Hopkins Hospital, No. 20, 

 March, 1892. 



2 Abstract, Science, No. 457, November 6, 1891. 



swollen, but not fatty. The liver was very fatty; the lighter 

 areas and dots were seen to correspond to foci of dead liver 

 cells, whose refractions were much greater than that of the 

 normal cells. 



Cultures were made from the blood and organs of the 

 animal, and they remained sterile. Cover-slips also were 

 examined and no organisms found. 



The histological lesions observed in this case are identical 

 with those described by us in counection with the inocula- 

 tions of the living organisms. Lymphatic apparatus: In 

 general, the changes are the same throughout. They are 

 found in the greatest intensity in the glands of the axillary 

 and inguinal regions, and less in the bronchial and cervical, 

 mediastinal, and mesenteric glands. Yet these are consid- 

 erably affected. The same fragmentation of nuclei affecting 

 the lymph- nodes and sinuses is met with. These fragments 

 exhibit the variety of form previously described by us, and 

 they have the same affinity for coloring agents. Much of 

 the nuclear detritus is free, but a part is contained within 

 large pale cells. In the spleen there is a similar diffuse 

 fragmentation of the nuclei of the spleen cells. Both the lym- 

 phoid cells of the follicles and the larger cells of the sinuses 

 are affected. Like the lymphatic glands, some of the nuclear 

 detritus ti enclosed in large cells. Besides the destruction of 

 cells in the spleen there is hemorrhage into the organ, or an 

 extreme degree of congestion, so that the tissue elements are 

 widely separated from one another. Nuclear figures occur 

 in the lymph glands and spleen. In the former they are 

 found among the fragmented cells. 



Stained sections of the liver, especially those stained in 

 methylene-blue and eosine, show the yellowish-white areas 

 to be composed of hyaline, necrotic liver cells. The necrotic 

 cells stain deeply in the eosine, and they are usually devoid 

 of nuclei. They form, on the whole, more or less definite 

 foci of hyaline cells into which leucocytes have wandered. 

 The largest area was two millimetres in diameter, and the 

 outlines of it were formed by hemorrhage into the tissues 

 corresponding with the hyperaemic zone spoken of above. 

 The cells in this focus have lost their nuclei and they are in- 

 tensely refractive. Many of the dead cells have retained 

 their individuality, and, indeed, their borders are more dis- 

 tinct than those of the normal cells. Others, however, tend 

 to become fused together and to lose their individual cell 

 outlines. Occasionally, outside the main focus of hyaline 

 cells, single necrotic cells occur, which are surrounded by 

 quite normal ones. Many leucocytes have wandered into 

 this area of dead liver cells, and they are especially abundant 

 at one place in the focus in which the hyaline cells are in pro- 

 cess of disintegration. An exquisite nuclear fragmentation 

 is to be observed throughout this area. 



Should the focus just described be compared to many 

 similar foci which occur in the livers of animals dead of in- 

 oculation with the bacilli themselves, it will be seen to con- 

 tain more polynuclear leucocytes within it. The explanation 

 of this fact would seem to depend somewhat on the inoculation- 

 time, but more, probably, on the progression, or stage, of the 

 necrotic process. Inoculation of the bacilli usually leads to 

 death in a very short time, twenty-four to forty-eight hours. 

 In this inoculation with the toxic products alone, the incu- 

 bation period exceeded three weeks. On account of this, 

 time has been allowed tor the softening and disintegration of 

 the dead cells, and leucocytes have been strongly attracted 

 to these foci. 



In the kidneys, besides the condition described in the frozen 

 sections, a slight fragmentation of the nuclei of the tubular 



