of growth fluctuated and gradually decreased. In the fifteenth passage the culture was divided and two cultures were made; one of the cultures (figure 2) was fixed after forty-eight hours' incubation. After the twentyfourth passage cell proliferation stopped. This culture was transferred twenty-five times, during a period of fifty-one days. The entire history is given in table I, which shows the passage, treatment, and observations that were made on culture 3.

Passage

Date [1913]

22

22

Jan. 4

Experiment 2.-A series of cultures, Nos. 1, 2, 3, and 4, was made from fragments of the periphery of a large round cell sarcoma3, and cultivated in equal parts of plasma and Ringer's solution. After twenty-four hours there was no evidence of growth in any of the cultures and the medium was in good condition. In forty-eight hours there was evidence of cell proliferation in all cultures. After seventy-two hours the area of cell proliferation had increased, but the medium in cultures 3 and 4 4 had become liquefied. Cultures 1 and 2 were stained.

Cultures 3 and 4 were changed into fresh medium (first passage), and one part of extract was added to the medium. Both cultures developed good growth in forty-eight hours with no liquefaction of the medium. They were again transferred into fresh medium (second passage). Culture 4 showed a few proliferating cells after seventytwo hours' incubation, and the medium was in good condition. The culture was discarded. After twenty-four hours' incubation in the second passage culture No. 3 showed growth with no liquefaction. In forty-eight and seventy-two hours growth had increased, but the medium was partially liquefied. The culture was transferred into fresh medium (third passage) and showed a few cells which had spread out into the medium from the central fragment. After seventy-two hours there was no increase in the number of cells, and the medium had liquefied. The culture was transferred into fresh medium (fourth passage), and after forty-eight hours good growth was observed. The medium was in good condition. The fifth passage into fresh medium was made in the same manner as in the previous passage. After twenty-four hours the medium was totally liquefied and no growth was observed. The fragment was transferred to fresh. medium, but after 24, 48, and 72 hours no growth developed. The culture was discarded.

RESULTS.

Two experiments were made in which fragments from human sarcomatous tissue were cultivated. It was possible to keep cultures of such tissue in a condition of active life in vitro for several generations. During the first twenty-four hours of incubation there was usually no evidence of cell proliferation, and slight liquefaction around the primitive fragments. When no liquefaction occurred, growth of new cells manifested itself after forty-eight hours. Twenty-four hours after passage into fresh medium (first passage), cell proliferation was observed in those cultures which showed no evidence of growth when first cultivated. In comparison with human connective tissue, the rate of growth was practically the same in the beginning, but a gradual decrease in the activity and extent of cell proliferation was observed as the length of time increased during which the culture was carried through successive 5. Pathological diagnosis: large round cell sarcoma.

passages. Microscopic examination of the first outgrowth of cells showed the presence of large, round, as well as elongated and ramified cells. In subsequent passages the round cells were no longer to be identified, and the elongated, ramified variety only were observed. The morphological characteristics of these cells did not appear to differ from the cells present

in cultures of normal human connective tissue. Preparations stained with Giemsa stain showed the large round cells as having a densely stained cytoplasm with from one to two nuclei and a regular outline. The elongated and ramified varieties showed no difference in comparison with those present in cultures of human connective tissue, with the exception that no mitotic figures were observed. Figure 3 shows a few

of the peripheral cells in a culture of sarcomatous tissue which had been carried through twelve passages.

One culture was stained which was growing actively in its twelfth passage (twenty-one days). This culture is shown in figure 1, the area of cell proliferation being that which developed in the twelfth passage during forty-eight hours' incubation. One other culture was carried through for twenty-four passages, that is, fifty-two days. It was possible to divide this culture in its fifteenth passage, making two, and after forty-eight hours; incubation one of these cultures was fixed. Figure 2 shows almost the entire culture.

Sarcomatous tissue grew as well during a few days as normal connective tissue. Afterwards the rate of growth became less rapid and the tissue could not be kept alive for more than fifty-two days, while normal human tissue could be kept for sixty-eight days.

These differences may be due to technical factors, but they may also be the result of the nature of the tissue itself. In his attempts at keeping Rous sarcoma in a condition of permanent life in vitro, Carrel observed that after a few generations the rate of growth became less rapid than the rate of growth of connective tissue. In other experiments with rat sarcoma and normal rat connective tissue, cultivated in guinea pig plasma, Carrel also observed the same difference. The writers observed the same phenomena when rat sarcoma and normal heart tissue of the rat were cultivated in chicken plasma.

The results obtained show that it is possible to cultivate in vitro fragments of human sarcomatous tissue for several generations, and that the method employed may prove of value in the study of the growth of human malignant tumor.

6. Carrel, A., Jour. Exper. Med., 1912, xv, 516.

SCOPOLAMINE-NARCOPHIN SEMINARCOSIS IN LABOR.*

By JAMES A. HARRAR, M. D. and ROSS Mc PHERSON, M. D. Attending Surgeons

UNDERTAKEN rather in a spirit of skepticism, the present investigation was begun by us several months ago. Doubtless many others have shared our recent experience in being the recipients of inquiries on account of sensational articles in the lay press on "painless child-birth. The first attitude was naturally to ridicule the whole matter as preposterous and to recall the agitation on the subject among obstetricians in 1907 and 1908, at which time the method was tried and found dangerous. Many foreign observers, notably Steffens,' Leopold, Hocheisen2

*Read at the Meeting of the American Association of Obstetricians and Gynecologists, Buffalo, N. Y., September 17, 1914. Appears also in Amer. Jr. Obst., Oct., 1914.