FREE AMINO ACIDS IN BLOOD. IV 397 
studied with a control sample followed by samples at 5, 24, 48, 72, and 96h. All of 
the animals in the 2.5 mg/kg group and 1 of the animals in the 1 mg/kg group were 
sacrificed at the end of 4 days since this was the time of maximum response of peri- 
pheral blood leukocytes to the mustard injection. The 2 remaining animals in the 
I mg/kg group were sacrificed at 9 and 13 days. The liver, kidneys, spleen, appendix, 
and bone marrow were removed from all animals and examined for free amino acids. 
The total leukocyte, granulocyte, and lymphocyte counts were determined for each 
blood sample and a micro-hematocrit value was obtained on each occasion. 
The effects of phenylhydrazine on the free amino acid levels of plasma and cells 
in rabbits was determined on blood samples taken from 3 rabbits that had been 
injected extensively with phenylhydrazine. These animals were supplied by Dr. 
Henry Borsook and Dr. GEOFFREY KEIGHLEY of the California Institute of Tech- 
nology. These animals showed a marked reticulocytosis that represented 88, 91 and 
94° of the circulating red cells, respectively, in the 3 animals. At autopsy the animals 
were seen to have marked changes in the liver and other organs. The animals were 
generally bled 1 or 2 days prior to the expected time of death where the maximum 
reticulocytosis was obtained. 
RESULTS AND DISCUSSION 
Effects of mtrogen mustard on free amino acids of blood in humans 
The results of the study of the administration of 4 doses of 0.7 mg of nitrogen mustard 
intravenously on 4 consecutive days to a patient with chronic lymphatic leukemia 
are shown in Figs. 133-144. Figs. 133, 134 and 135 show plasma, erythrocytes and 
leukocytes examined just prior to injection of the first dose of nitrogen mustard. The 
plasma glutamine level was somewhat lower than normal and the plasma glutamic 
acid level was distinctly above the normal level in the control sample. The total 
erythrocyte free amino acid pool was above normal, while the leukocyte free amino 
acid pool was most notable for the very low level of glutamine (see discussion in 
part III). 
Figs. 136, 137 and 138 show the plasma, erythrocytes and leukocytes, respectively 
on the third day of the study after a total dose of 1.4 mg of nitrogen mustard had 
been administered. The plasma amino acid levels were higher with the most noticeable 
increases seen for the leucines, valine, glycine, glutamic acid and aspartic acid. 
Taurine was decreased. The erythrocyte free amino acid pool showed a general 
decrease in free amino acids with the most marked change being a reduction in the 
glutathione level. The total free amino acid content of the leukocytes was increased 
with particularly large increases in taurine, valine and leucine plus isoleucine. 
Figs. 139-141 show the plasma, erythrocytes and leukocytes obtained 1 day after 
the total dose of 2.8 mg of nitrogen mustard had been administered (the fifth day 
of the study). The plasma levels of the leucines and valine resembled the control 
levels and taurine was much nearer the control level. Most noticeable was the marked 
decrease in glutamine and the high level of glutamic acid (seen also in the previous 
plasma sample). The free amino acid pool of the erythrocytes shown in Fig. 140 re- 
presented a decrease of all free amino acids to a level even lower than seen in the 
previous blood sample. Glutathione and taurine were completely absent from the 
erythrocytes. As shown by Figs. 134, 137 and 140 there was a progressive drop in 
References p. 447/448 
