430 OCCURRENCE OF FREE AMINO ACIDS —- VERTEBRATES 
FREE AMINO ACIDS IN THE BLOOD OF MAN AND ANIMALS 
VI. CHANGES FOLLOWING GLUTAMINE INGESTION 
BY NORMAL INDIVIDUALS AND PATIENTS WITH CHRONIC LEUKEMIA 
GEORGE ROUSER, KEITH KELLY, BOHDAN JELINEK, EUGENE ROBERTS 
AND FRANCIS W. SAYRE 
Departments of Biochemistry and Medicine, City of Hope Medical Center, 
Duarte, Calif. (U.S.A.) 
These studies arose from the observation that the patients with chronic leukemias 
tended to have a somewhat lower than normal plasma glutamine level and that 
some of the leukocytes appeared to metabolize glutamine relatively rapidly. It was 
then decided to investigate whether or not glutamine might be a limiting metabolite 
in these patients by observing the effects of ingestion of a large load of this amino 
acid. 
GENERAL METHOD OF STUDY 
The subjects were fasted overnight, I to 3 control blood samples and at least one 
urine sample were obtained; the subjects then ingested 50 g of L(-+)-glutamine, 
and were followed at intervals up to 8 h. Following the ingestion of amino acid there 
was no further intake of food, but a limited intake of water was allowed in some 
cases. 
Three adult males on the research staff were studied as controls (a total of 26 blood 
samples and 12 urine samples obtained). One patient with chronic lymphatic leuke- 
mia (R. Tho.) without any prior form of treatment was studied (13 blood samples 
and 5 urine samples obtained). The response of this same patient to dimethylmyleran 
is described in part V (see Figs. 195-230). Three patients with chronic granulocytic 
leukemia were studied before treatment and one of the patients (H. Gol.) was studied 
on two occasions after treatment with dimethylmyleran. The other two patients 
(R. Tap. and Q. Tur.) were studied by paper chromatography and glutamine levels 
of plasma and cells were determined with an enzymatic assay. The method of SAYRE 
AND RoBerts”® was used for the preparation of the purified enzyme. An 80% ethanol 
extract was prepared, evaporated to dryness, and an aliquot in buffer used for assay. 
A total of 40 blood samples and 12 urine samples were examined from the granulocytic 
leukemia patients. A total of g glutamine ingestion studies were carried out (a total 
of 79 blood samples and 29 urine samples examined). 
The blood samples from the normal individuals were centrifuged at 1300 * g for 
8 min, the plasma aliquot withdrawn, the white cell layer overlying the red cells was 
removed, and erythrocytes were then removed largely free of contamination with 
leukocytes and platelets. The samples were then extracted. The processing of the 
blood of the leukemic subjects was as described in part I. Blood samples from 
normal individuals required 15~20 min for processing (from the time of withdrawal 
of blood to extraction), while the blood samples from the patients required 30-35 min 
for workup. 
References p. 447/448 
