316 E. ROBERTS AND D. G. SIMONSEN 
Results similar to those above (Figs. 169-180) were obtained when 3.5 mg of 
sarkomycin was administered to 35-g mice 48h after inoculation with the Ehrlich 
ascites tumor. Even at 5 min after the injection both cytoplasmic and nuclear damage 
were evident (Fig. 172) and glutamine, which was not detected in the control cells 
or fluid, appeared (Figs. 173, 174). There was a marked decrease in the content of 
glutathione in the tumor cells. Valine and the leucines increased in amounts, and a 
new ninhydrin-reactive material, possibly an addition product of glutathione and 
sarkomycin, appeared to the left of glutamic acid (spot X) and persisted through 
the period of study. The chromatographic results obtained at the ro- and 20-min 
intervals (results for ro min, Figs. 173, 174) were similar to those found at the 5-min 
period with the exception that taurine appeared in the fluid in considerably larger 
amounts. However, the cytological damage progressed rapidly (Fig. 175). The cells 
in the 40-min sample (Fig. 178) showed extensive cytoplasmic and nuclear damage. 
In many cells disintegration of nucleus and cytoplasm appeared to be occurring 
simultaneously. The ascitic fluid (Fig. 180) showed elevations in the levels of gluta- 
mine, taurine, and aspartic acid over the control. At this time, however, the badly 
damaged cells (Fig. 179) showed markedly elevated levels of glutamine, valine, 
leucine and isoleucine, tyrosine and lysine over the controls, while the other ninhydrin- 
reactive constituents were unchanged. It is truly remarkable that cells showing the 
degree of destruction observed in Fig. 178 should be able to retain high levels of 
small molecules such as amino acids. The findings are certainly not in keeping with 
these constituents being in free solution in the cytoplasm or nucleoplasm and being 
maintained intracellularly by intact membranes. Separate experiments with 'C- 
labeled glutamine gave results in keeping with the interpretation that the effect of 
sarkomycin resulting in the increased content of easily extractable glutamine in the 
tumor cells is not necessarily related to an altered ability of the tumor cells to take 
up exogenous glutamine and to convert it to glutamic acid*4. 
Podophyllin is another material which produces both cytoplasmic and nuclear 
damage in tumor cells. Podophyllin (10-15 mg/kg) was administered to J-strain 
rats bearing the Yoshida sarcoma. Extensive and progressive damage occurred to 
the tumor cells even within 1 h after the administration of the drug, many irregular 
processes or atypical ameboid protrusions appearing in the cytoplasm of the cells 
and all cells in metaphase showing chromosome clumping or other degenerative 
changes such as formation of deformed, rounded, or bizarre bodies. The chromatog- 
rams of both extracts from the cells and the fluid for the control sample and for 
those obtained at 30, 45 and 60 min after injection are shown in Figs. 181-188. In 
the case of the ascitic fluid, the post-injection samples showed the presence of taurine, 
glutamic acid, aspartic acid, and ethanolamine phosphate in small but detectable 
amounts, while these constituents were not noted on the chromatograms of the con- 
trol sample of fluid, suggesting a slight leakage of some of the intracellular constit- 
uents. At most, traces of glutamine were detected in the extracts of fluid or cells 
at the times studied. The most remarkable thing about the above findings is the 
constancy of the pattern of the easily extractable amino acids of the tumor cells 
even at 60 min after treatment (compare Fig. 187 with the control in Fig. 181) at 
a time when marked cytological damage had occurred. A similar constancy in amino 
acid distribution in cells and fluid was found in a serial study of the Yoshida tumor 
after treatment with nitromin (methyl-bis-($-chloroethyl)amine-N-oxide hydro- 
kreferences p. 348/349 
