ALBERT

Description: 1. Two morphologically distinct populations of erythrocytes were found in a mother and in two of her daughters, one of whom (the proband) was anemic. One erythrocyte population was morphologically normal; the other was hypochromic and microcytic. 2. The X-linked blood group antigen, Xga, was present in erythrocytes from the mother and the two daughters, but not in erythrocytes from the father. The daughters were, therefore, heterozygous for the gene controlling this antigen. 3. Separation of the two populations of erythrocytes was accomplished by centrifugation in layered gum acacia solutions of different specific gravity. 4. The microcytic cells from the three affected individuals were Xga-positive. Isolated normal cells were Xga-positive in the mother, but negative in both daughters. These data suggest that the erythrocyte defect is X-linked and that the phenomenon of X-inactivation applies to genes controlling both the morphologic defect and the Xga antigen. 5. The free protoporphyrin content of the isolated microcytes was lower than that of the normal cells. The capacity of the microcytes to convert delta-aminolevulinic acid to protoporphyrin was unimpaired. On these bases it is suggested that the hereditary defect lies either at or before the step in which delta-aminolevulinic acid is synthesized.

Description: Studies are described which deal with the metabolism of irons in swine made anemic by feeding a milk diet low in copper. Evidence is presented that in such animals there is (1) impaired ability to absorb irons from the gastro-instestinal tract; (2) incomplete mobilizations of irons from the tissues; and (3) inability to utilize parenterally administered irons for hemoglobin synsthesis evens when it is presented to the bone marrow in normal quantities.

Description: Present knowledge concerning the kinetics of granulopoiesis has been reviewed and quantitative data concerning granulokinetics in normal human subjects are presented. A. When granulocytes are labeled in vitro and returned to the circulation of the donor, the distribution of the cells in the circulation and the rate of disappearance of the cells from the circulation can be measured. 1. The total blood granulocyte pool (TBGP) consists of two compartments which are in equilibrium with each other. These pools have been designated the circulating granulocyte pool (CGP) and the marginal granulocyte pool (MGP). The size of the pools has been measured in 109 normal male subjects. The mean values, expressed as numbers of cells x 107 per Kg. of body weight were as follows: TBGP, 70; CGP, 31; and MGP, 39. The mean ratio of the CGP to the TBGP was 0.44. 2. The labeled granulocytes leave the TBGP in an exponential fashion with a mean half-time disappearance (T½) of 6.7 hours as determined in 56 normal male subjects. No evidence has been obtained for a return of granulocytes to the blood. 3. The mean value for the granulocyte turnover rate (GTR) in 56 normal male subjects was 163 x 107 granulocytes per Kg. of body weight per day. Thus, the TBGP turns over 2.3 times per day and the turnover time for the TBGP is 10.4 hours. B. When granulocytes are labeled in vivo by the intravenous administration of DFP32, the rate of disappearance of granulocytes from the circulation and the time required for myelocytes to divide, mature and appear in the blood can be measured. In addition, the generation time of myelocytes can be approximated. From the time parameters and the GTR, the bone marrow pool sizes and turnover times can be calculated. These determinations and calculations have been made for a group of 21 normal male subjects. 1. The mean half-time disappearance (T½) of in vivo labeled granulocytes from the circulation was 7.2 hours. This value agrees well with the value of 6.7 hours obtained after the in vitro labeling of granulocytes. 2. The mean time required for myelocytes to divide, mature and appear in the blood was 11.4 days. 3. The mean generation time of myelocytes was estimated to be not more than 2.9 days. 4. The total granulocyte pool in the bone marrow (neutrophilic myelocytes, neutrophilic metamyelocytes and PMN neutrophils) was calculated to be 186 x 108 cells per Kg. of body weight with a mean turnover time of 11.4 days. The myelocyte pool was estimated to be 41 x 108 cells per Kg. with a turnover time of 2.5 days; the metamyelocyte pool consisted of about 76 x 108 cells per Kg. with a turnover time of 4.7 days; the average size of the mature marrow PMN neutrophil pool was 69 x 108 cells per Kg. of body weight with a turnover time of 4.2 days. C. A kinetic model for granulopoiesis, based on the studies with the DFP32 label, is presented. In this model, myelocytes are depicted as approaching a self-perpetuating population of cells. Some cells enter this population from populations which are less mature but this latter source of cells is small under conditions of normal steady state kinetics. One of the daughter cells of a myelocyte division remains in the myelocyte population to divide again. The other daughter cell enters the metamyelocyte population. The metamyelocyte and PMN neutrophil population is incapable of division and cells move through this population in sequential fashion in the process of maturation. The cells then enter the blood where they equilibrate rapidly between the two blood compartments. The cells are removed from the total granulocyte pool in a random fashion. There is no appreciable pool of granulocytes in the extramedullary tissues of normal subjects and granulocytes do not return from the tissues to the blood. The entire movement of granulocytes from marrow to tissues is uni-directional.

Description: A 26 year old American housewife of Sardinian extraction with chronic hypochromic anemia was found to have a hemoglobin component identical to hemoglobin H. The A2 hemoglobin fraction was decreased. Relatives were found to exhibit the hematologic features of thalassemia trait. In this pedigree, in contrast to the usual finding in the "thalassemia trait," the A2 values were not increased. The same observation had been made in the only other comparable report.

Description: 1. The urinary excretion of uric acid was studied in 17 normal subjects and in 38 patients with leukemia. The mean excretion of uric acid by the normal subjects was 6.5 mg./Kg. of body weight /24 hrs. The mean excretion of uric acid in 14 patients with acute lymphoblastic leukemia was 30.3 mg./Kg. /24 hrs.; in 13 patients with acute myeloblastic leukemia, 13.0 mg.; in 6 patients with chronic lymphocytic leukemia, 5.2 mg.; and in 5 patients with chronic myelocytic leukemia, 13.5 mg. 2. Following therapy with cortisone, 6-mercaptopurine or Amethopterin, the urinary excretion of uric acid increased in the cases of acute leukemia as the leukocyte count declined. As the leukocyte count approached normal levels, the uric acid excretion decreased. The urinary excretion of xanthine and guanine paralleled the excretion of uric acid. 3. In association with the administration of an aromatic nitrogen mustard derivative to one patient with chronic lymphocytic leukemia and Myleran to one patient with chronic myelocytic leukemia, there was only a slight increase in uric acid excretion.

Description: Studies of a patient with acquired sideroachrestic anemia and metastatic carcinoma of the prostate are presented. Although the administration of pyridoxine did not elicit a hematologic response, the administration of this vitamin was associated with a remarkable accumulation of protoporphyrin within the erythrocytes. It is suggested that there were two defects in the hemoglobin biosynthetic pathway: impaired incorporation of protoporphyrin into hemoglobin and defective delta-aminolevulinic acid synthesis. Pyridoxine had no effect on the former hut partially corrected the latter.

Description: A total of 20 swine were fed a diet adequate in all known respects except that soybean protein was substituted for casein, succinylsulfathiazole and a folic acid antagonist were added, and vitamin B12 and pteroylglutamic acid were withheld from the vitamin supplement. The animals developed macrocytic anemia, leukopenia and neutropenia, accompanied by erythroid hyperplasia of the bone marrow. Tue erythroblasts consisted mainly of immature macronormoblasts but a few atypical megaloblasts were also observed. The anemia responded rapidly and completely to the administration of both vitamin B12 and pteroylglutamic acid. The administration of pteroylglutamic acid alone resulted in an immediate return of the blood and bone marrow to within normal limits but after several months there was a partial hematologic relapse in spite of continued therapy with this vitamin. The administration of vitamin B12 alone resulted in only partial remission of the anemia and the bone marrow remained macronormoblastic although the megaloblasts tended to disappear. Growth of the animals was stimulated by the administration of either vitamin but the administration of both vitamins simultanseously resulted in the greatest rate of growth. No manifestations of neurologic disturbances or of inscreased pigment excretion were observed in the deficient swine.

Description: The intraperitoneal administration of methyl cellulose into rats over a period of fifteen weeks resulted in the development of a syndrome characterized by massive splenomegaly, hyperplasia of the bone marrow elements, normocytic, normochromic anemia, reticulocytosis, leukopenia, a mild thrombocytopenia in 9 of the 10 animals, ascites, and infiltration of the spleen, liver and kidneys with "storage-cell" macrophages. The administration of methyl cellulose to rats previously splenectomized produced similar histologic lesions but failed to produce the hematologic abnormalities. It is suggested that the syndrome produced by methyl cellulose may represent an experimental form of the process to which, in man, the term "secondary hypersplenism" has been applied.

Description: 1. Following splenectomy in the albino rat, the total leukocyte count increased approximately 100 per cent in seven days, and remained significantly elevated for seventy to ninety days, after which time time leukocytes returned to normal levels. This increase in circulating leukocytes was due to an increase in both neutrophils and mononuclear cells. Partial omentectomy and unilateral nephrectomy produced increases of less magnitude and much shorter duration than those which followed splenectomy. 2. Removal of as much as 75 per cent of the spleen resulted in a leukocyte increase resembling in magnitude and duration that of control operations. 3. When small portions (less than 10 per cent) of the spleen were transplanted to other sites, the leukocyte response also resembled that which followed the control operations. 4. When splenectomy was performed in one partner of parabiotic rats, no rise occurred in the leukocyte count of either animal. When the spleen of the second partner was then removed, a rise in the leukocyte count of both animals occurred. 5. When rats were made leukopenic by pteroylglutamic acid deficiency, no rise in the leukocytes in the peripheral blood occurred following splenectomy. 6. It is concluded that in the rat the spleen exerts an influence on the level of circulating leukocytes, and that the results of these experiments support but do not conclusively prove the hypothesis that this organ exerts this influence by controlling the rate of production and/or liberation of leukocytes in the bone marrow. These studies do not exclude the possibility that under certain circumstances the spleen may destroy white cells.

Description: In an effort to produce a deficiency of vitamin B12 a total of 70 pigs were fed a purified diet containing soybean alpha protein in place of casein. One group of animals was started on the diet at 2 to 7 days of age. A second group began at 21 to 28 days of age. Methionine, iodinate casein, desiccated thyroid and pteroylglutamic acid were added to the diet of certain animals and! omitted from the diet of other pigs. In addition, 9 pigs were gastrectomized. Forty-three of the animals survived for a sufficiently long period of time for adequate evaluation of the results of the experiment. Severe liver damage was observed in 24 of the 25 animals autopsied. The only animal not showing liver damage received vitamin B12 from the beginning of the experiment. Necrosis of the liver cells, fatty infiltration, or both, occurred in the presence of a high fat diet containing apparently adequate amounts of protein, choline, vitamin E and methionine. These pathologic changes were apparently prevented but not reversed by the administration of vitamin B12. Growth of the animals on the above diets without added vitamin B12 was retarded as compared with the growth of animals on the same diet supplemented with this vitamin. The administration of vitamin B12 to the deficient animals resulted in rapid growth. Of the 39 animals not receiving vitamin B12 13 failed to develop anemia, 16 developed a mild anemia and in 10 a moderately severe anemia was present. When present the anemia was normocytic and in 24 pigs was accompanied by a moderately severe neutropenia. Differential cell counts on the sternal marrow were normal except for a slight increase in the proportion of normoblasts. These hematologic alterations were neither consistently or completely corrected by the administration of vitamin B12 in spite of the fact that definite and sometimes marked reticulocyte increases followed. When methionine deficiency was associated with vitamin B12 deficiency, anemia appeared to be more severe. The administration of aureomycin, an "animal protein factor," did not stimulate growth and failed to induce a hemopoietic response. There was no macrocytic anemia, the bone marrow was not megaloblastic, and neurologic disturbances or morphologic alterations in the neutrophils were not observed. These results are in contrast to those obtained in pigs with an experimentally produced deficiency of pteroylglutamic acid. Such animals develop macrocytic anemia, leukopenia and a macronormoblastic type of bone marrow. It is not possible to give with any assurance the reason why megaloblastic anemia was not produced in the "B12-deficient" animals. This may have been due to the fact that (1) the deficiency was not sufficiently severe to result in such a change in the hemopoietic system; or (2) because pteroylglutamic acid prevents the development of megaloblastic anemia even in the absence of vitamin B12.