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Researches on the cultivation of the parasites of malaria in Liverpool were commenced some time ago at my suggestion by Dr. Sinton, and then, with better success, by Drs. J. G. Thomson and McLellan, and by Dr. D. Thomson. We are greatly obliged to Sir Edwin Durning-Lawrence, Bart., for giving us the services of Dr. J. G. Thomson for this important enquiry.—Ronald Ross, 21st May, 1913.

Prefatory Note by R. Ross ―For a long time it has appeared to me that much light might be thrown on infectious diseases, immunity and treatment, by more exact enumeration of the infecting organisms, and that we might even be able ultimately to apply mathematical reasoning to the study of these subjects. In 1903 I elaborated a method of blood examination, called the thick-film process , which enables us to detect small organisms in the blood about twenty times more easily us to detect small organisms in the blood about twenty times more easily than in ordinary preparations; but for the lack of the necessary assistance I was long unable to apply the method to the laborious enumeration of such organisms. Recently Fund has placed considerable funds at the disposal of the Liverpool School of Tropical Medicine for the study of cases in the tropical ward of the Royal southern Hospital, Liverpool―with the result that the investigations referred to were commenced by Dr. David Thomson and myself from the beginning of this year. As I expected, methodical counting of the parasites has at once verified or disclosed several facts of importance in connection with malaria and trypanosomiasis. We now limit ourselves to a brief description of the remarkable periodical increase of Trypanosoma gambiense revealed by careful daily counting in a case in my clinic (the case, which is still under treatment, will be fully reported elsewhere). W. A., male, aged 26 years, a strong young man born in Northumberland, was infected in N. E. Rhodesia in September, 1909, the trypanosomes being found in his blood in Africa on November 17. On coming to Liverpool for treatment he was admitted into the Southern Hospital on December 4. From then until February 16 (73 days) the number of trypanosomes in his blood was estimated only by the rough methods in common use; that is, by the proportion of trypanosomes to red cells or leucocytes, or to “fields” of the microscope examined. These methods are obviously open to such great error that they can scarcely be depended upon to indicate any but very large differences in the numbers of objects counted. During the 73 days 46 counts were made; but on several occasions none was attempted for three or four days in succession―so that, even if the methods of counting employed had been more accurate, sudden fluctuations might easily have been missed. Hence, as was to be expected, the graph during this period is very irregular and almost worthless. On admission on December 4 the patient was reported to contain about 6000 trypanosomes per cubic millimetre, were found on December 17 and 28 and on January 16. All this time the patient was given the usual treatment with atoxyl and mercury, and received altogether 10 doses of two to four grains of the former. Nevertheless the parasites never fell below about 200 per cubic millimetre in number, as roughly estimated.

Prefatory Note by R. Ross .—Towards the end of last year the Advisory Committee for the Tropical Diseases Research Fund (Colonial Office) allotted considerable funds to the Liverpool School of Tropical Medicine for researches to be carried out in Liverpool. This enabled us to commence, under my direction, a number of minute co-ordinated studies on cases admitted into the Tropical Ward of the Royal Southern Hospital—material which, though it offered pecuilar facilities for research, had long remained neglected owing to want of funds. The cases (occurring in the clinics of Dr. Macalister, Dr. Lloyd Roberts, and myself) were placed in charge of Dr. David Thomson; the chemical studies in charge of Dr. G. C. E. Simpson; the parasitological studies in charge Dr. H. B. Fantham. Parallel researches on animals were also assigned to Dr. John Thomson who is working under Sir Edward Durning-Lawrence’s fund the investigation of the effect of temperature on disease; Dr. V. T. Korke (Research Fellow) has studied coagulation times and other details; the literature was in charge of Mr. W. R. Drawz, the Malaria Bibliographer (Advisory Committee’s Fund); and much valuable help has been given by the staffs of the University, the School of Tropical Medicine, and the Royal Southern Hospital of Liverpool, and by Sister Linaker of the Tropical Ward. The researches were commenced on January 1, 1910. A paper by Dr. David Thomson and myself, describing a regular periodical increase of the trypanosomes in a case of Sleeping Sickness, was published; and we now present to the Society brief accounts of our results regarding malaria, blackwater fever, trypanosomiasis, and various therapeutical agencies, obtained (mostly by new methods) up to the end of July, 1910. Further details will be published, if necessary, in the ‘Annals of Tropical Medicine,’ Liverpool.

In a previous paper by us, published in the ‘Proceedings’ of July 21, 1910, B 557, we recorded our observations on this case during two and a-half months, and described particularly the regular periodical rises in the numbers of trypanosomes disclosed in the patient’s peripheral blood by methodical daily countings extending over that period. Our technique has been detailed in another paper by us on “Enumerative Studies on Malarial Fever,” recently submitted to the Society. We now record our further observations on the case during two more months—until the patient’s death. A chart and a table giving daily details of the observations are attached; and accompanying papers by Drs. J. G. Thomson and H. B. Fantham record studies on animals and on the parasites themselves. We must refer also to a recent communication to the Society by Drs. J. W. W. Stephens and Fantham suggesting that the species found in this case may not be identical with T. ganibiense . The patient, a strong young Englishman, age 26, weight 154 lbs., was infected in N. E. Rhodesia near the River Luangwa in September, 1909. The trypanosomes were found in his blood in Africa on November 17.

On two occasions the authors have definitely succeeded in cultivating human tumour tissue in vitro . The tissue was obtained at operations performed by Sir John Bland-Sutton at the Middlesex Hospital, and conveyed in sterile Ringer’s solution in a thermos flask to the laboratory, where small portions were immediately inoculated into the culture medium. (a) ' Intracystic Papilloma of the Ovary (not truly malignant).—This tissue was grown in a medium composed of fowl plasma 1 part, Ringer’s solution (containing 0·5 per cent, of glucose) 1 part, and extract of the tumour in Ringer’s solution 1 part. On the third day of incubation at 37·5° C. definite buds of new growing tissue appeared. On the fifth day these were more distinct and on the eighth day the amount of growth had increased considerably (fig. 1, Plate 7). This growth consisted of a solid extension of epithelial cells. As the growth increased it caused some liquefaction of the medium, which was of a gelatinous consistence, and in the more liquefied parts the new growing cells were scattered (fig. 2), but as a rule they remained in contact with each other by means of long fine protoplasmic connections (fig. 3). The new actively proliferating cells varied markedly from the cells of the original tissue planted in the medium. The former were large and amoeboid, with long processes which communicated with each other, and they also contained large highly refractile granules. The original cells, on the other hand, were much smaller; they showed no amoeboid processes, did not exhibit amoeboid movement and they contained few or no refractile granules. This tumour was a very soft one and appeared to contain little or no fibrous stroma. It was composed entirely of epithelial cells, and it will be noted that the new growth also consisted of epithelial cells only.