Fairley (1) proved by the results of glucose tolerance tests, and Barker and Rhoads (2) by fat tolerance tests, that the absorption of sugar and fat by patients with sprue is improved after treatment with liver extract. Moreover, Groen (3) observed that the diminished rate of absorption of glucose by the intestinal tract of 3 patients with pernicious anemia became normal after similar therapy.From these observations it was concluded that in both sprue and pernicious anemia a dysfunction of the intestinal tract is present which may be corrected by some constituent of liver extract. These conclusions were not entirely warranted, however, since no proof was advanced that simple diarrhea, or ischemia of the intestine, unrelated to liver extract deficiency, would not result in a similar interference with absorption. Moreover, the evidence concerning the malabsorption of glucose and of fat is repeatedly advanced as proof that the effective principle of liver extract is also poorly absorbed. No adequate experimental data bearing on this point are at hand, however, since the studies of Dakin, Ungley, and West (4) and of Subbarow, Jacobson, and Hartfall (5) indicate that the active substance is a breakdown product of protein, probably a peptide. Accordingly, studies have been made of the absorption of an amino acid, glycine, by patients with sprue and pernicious anemia, as well as with other disorders. The results are presented in this communication.The glycine tolerance test was employed to obtain information concerning both the rate of absorption of the amino acid and the speed with which it is metabolized. Studies were made before and after the intramuscular administration of liver extract to 2 patients with sprue and to 5 patients with pernicious anemia. Twelve other patients-3 with pernicious anemia in induced remissions, 4 with ulcerative colitis, 3 with refractory anemia, and 2 with hepatic cirrhosis-were subjected to single tests and served as control cases.The tests were made by the measurement of the levels of the plasma amino nitrogen and the plasma and urinary urea nitrogen before and after the ingestion of glycine. The levels of the plasma amino nitrogen are considered to indicate the rates of absorption and deaminization of the amino acid, whereas the levels of the plasma and urinary urea nitrogen indicate the rates of urea formation and excretion. Kirk (6) has reviewed the evidence that most, if not all, of the urea in the blood is formed in the liver, and that absorbed amino acids are deaminized by that organ as well as by the kidneys. He also showed by the results of glycine tolerance tests that patients with acute uremia are not able to deaminize glycine as rapidly as normal individuals or as patients with chronic uremia.Heath and Fullerton (7) studied the levels of the plasma amino nitrogen after the oral as well as the intravenous administration of glycine. They concluded that "the amino acid nitrogen content of the blood following the ingestion of 25 grams of glycine gives no useful information reg...
Previous studies (1, 2) have shown that the phosphorus metabolism of mouse tissues is markedly altered by leukemic infiltration, the absorption and retention of radio-phosphorus being increased in the tissues of leukemic animals. The present work goes one The leukemic animals were fed Purina dog chow ad lib. The weighed daily food consumption of each group of leukemic animals determined the amount fed to its respective group of control animals on the following day. In addition, each group of five animals received six grams of whole oats per day and ten grams of fresh lettuce ,every other day. Thus the phosphorus and caloric intake of the leukemic and control animals was approximately the same.The radio-phosphorus was administered by intraperitoneal injection of 0.5 cc. of a solution containing 7.5 mgm. sodium phosphate at pH 7.4 and emitting 5.5 microcuries1 of beta activity at the time the first group was injected. Ten leukemic and ten normal animals were given this solution seven, five, three and one-half, two, and one-quarter days before all the animals were killed, which occurred on the eighteenth day after inoculation with leukemic cells. At this time the weights of leukemic and control animals were approximately the same.At autopsy, all leukemic animals had peritoneal tumors weighing about 800 to 1,000 mgm. and had uniformly enlarged spleens, livers and lymph nodes-both central and peripheral. These tissues (tumor, liver, spleen and lymph nodes) from the animals of each group were pooled, weighed and prepared for analysis. The weighed tissue was ground thoroughly under 95 per cent alcohol with the aid of a little sand and transferred to 100 cc. centrifuge tubes. The carcasses of each normal and leukemic group were pooled, ashed and assayed for radioactivity. METHODS OF TISSUE FRACTIONATIONA. Phospholipid. The tissue phospholipid was removed by four successive extractions, the first with 95 per cent alcohol and the other three with 3:1 alcohol: ether. Seventy cc. of solvent were used in each extraction which was carried out at 50 to 550 C. under a reflux condenser. The tubes containing the tissue were centrifuged after each extraction and the solvent decanted. The total duration of extraction was thirty-six hours.The combined alcohol and ether extracts were evaporated at low temperature to a volume of about 2 cc. Five cc. of water were added and the mixture transferred to a separatory funnel where it was extracted five times with 70 cc. portions of petroleum ether. The combined petroleum ether extracts were evaporated down to a small volume, transferred to ashing capsules, ignited, and set aside for analysis. The aqueous phase which contained only a small amount of activity was added to the acid soluble fraction, since it contained some of the simpler non-lipid compounds removed from the tissue in the course of the alcohol extractions.
The purpose of this paper is (a) to indicate the amount of radio-phosphorus (P32) retained by various fractions of the blood of 4 normal individuals, 12 patients with myeloid, and 15 with lymphoid leukemia, and the variations in retention following the administration of radio-phosphorus when given orally and/or intravenously and when accompanied by varying amounts of non-radioactive phosphorus (P31); (b) to indicate the distribution of pS2 in the bone marrow and in various fractions of white blood cells; and (c) to indicate the amount of radio-phosphorus excreted in urine and feces in these cases. MATERIALS AND METHODSThe radioactive phosphorus was produced by the Berkeley cyclotron (1). The 4 normal individuals were robust, ambulatory workmen with recently healed fractures, all of whom had received the same type and quantity of food over a period of from 1 to 8 weeks, and each of whom had a single regular bowel movement daily during the same period previous to the administration of P'. It was impossible to control the diets or the time of excretion of the patients, all of whom were ambulatory. The blood withdrawn from veins was heparinized, cooled and centrifuged exactly 20 minutes at 1450 times gravity to insure constant volume. The buffy coat was aspirated, suspended in equal amounts of heparinized Ringer's solution and centrifuged exactly 20 minutes at 1450 times gravity. The plasma was then removed from the original tube and finally the red blood cells were extracted. Bone marrow obtained by sternal aspiration was heparinized and centrifuged, as described previously. In some instances, the nuclei were separated from the cytoplasm of the peripheral white blood cells by violent agitation for 20 minutes in cold 5 per cent citric acid (2). The phospholipids were extracted from the white blood cells by the use of ether, alcohol and reflux condensers, and the acid-soluble substances by use of ice-cold 5 per cent trichloracetic acid, thereby leaving 1 These investigations have been aided by a grant from the John and Mary R. Markle Foundation.2Wm. R. Kenan, Jr., Fellow.the nucleoprotein-like substances as residue (3). The assays of radioactivity were made by use of an electrometer.RESULTS
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