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Heme synthesis by copper-deficient cells was investigated to elucidate the nature of the defect in intracellular iron metabolism. Iron uptake from transferrin by copper-deficient reticulocytes was 52% of normal, and the rate of heme synthesis was 33% of normal. Hepatic mitochondria isolated from copper-deficient animals were deficient in cytochrome oxidase activity and failed to synthesize heme from ferric iron (Fe III) and protoporphyrin at the normal rate. The rate of heme synthesis correlated with the cytochrome oxidase activity. Heme synthesis from Fe(III) and protoporphyrin by normal mitochondria was enhanced by succinate and inhibited by malonate, antimycin A, azide, and cyanide. It is proposed that an intact electron transport system is required for the reduction of Fe(III), thereby providing a pool of ferrous iron (Fe II) for protoheme and heme a synthesis.

The biphasic shape of the survival curve of 99 patients with aplastic anemia suggested that there may be at least two subgroups of patients with this disease, one with a very short survival and another with a longer survival. Patients who survived for 4 mo or less after the first clinic visit (group A) were different from the patients who survived longer (group B) with respect to their modes of onset, sex, intervals from the onset of symptoms to first clinic visit, and initial hematologic values. These differences suggested that short survival could be predicted from data available at the first contact with the physician. From these measurements, a prognostic index could be calculated which was useful in identifying the patients in group A. Although this method of prognostication needs further testing, if validated, it may prove useful in selecting patients for therapeutic trials and could explain the divergent results in previous studies of androgen treatment of aplastic anemia. When our androgen-treated subjects were compared with subjects with a similar prognostic index who had not received androgens, a beneficial effect of androgen therapy on survival could not be demonstrated.

The azide-resistant ferroxidase activity of fresh, undialyzed human serum, either normal or from patients with Wilson's disease, can be accounted for by citrate and by the incomplete inhibition of ceruloplasmin ferroxidase activity in 1 mM concentrations of NaN3. Prolonged dialysis of serum at pH 5.5 resulted in the development of appreciable ferroxidase activity when the dialyzed serum was assayed at pH 6.0 in acetate buffer. This activity could not be accounted for by either citrate or ceruloplasmin and corresponds to the ferroxidase II activity reported by others. Since this activity was not detected in human serum under physiologic conditions and its activity was a function of the pH, duration and temperature of dialysis, its physiologic role in iron metabolism has not been established.

This report summarizes information obtained on four generations of a kindred afflicted with a mild hemorrhagic diathesis. Studies carried out on a family of eight members and platelet counts obtained on fifty-one additional members demonstrated only mild thrombocytopenia in eleven members of three generations. The thrombocytopenia appears to be inherited as an autosomal dominant trait.

1. The amount of free coproporphyrin and protoporphyrin its the erythrocytes of twenty normal male subjects was found to be 0.76 ± 0.26 and 31.3 ± 11.9 µg./100 ml. of packed RBC, respectively. In ten normal female subjects the values were 0.71 ± 0.23 and 44.6 ± 19.6 µg./100 ml. of packed RBC, respectively. 2. In eighteen male patients with chronic infection, mean values of 1.53 ± 0.61 µg./100 ml. of packed RBC for free erythrocyte coproporphyrin and 181.2 ± 90.6 µg./100 ml. of packed RBC for free erythrocyte protoporphyrin were found.

SummaryThe chronic, disseminated inflammatory reaction observed in rats following a single intradermal injection of Freund's-type adjuvant was associated with a mild, non-progressive anemia which morphologically, biochemically and kinetically resembled the anemia associated with chronic disorders in man. It is suggested that “adjuvant disease” in the rat is an appropriate experimental model in which to study the complex pathogenesis of this distinctive but not uncommon form of anemia in man.

SummaryThis study was designed to determine whether inflammatory granulocytes are derived solely from the blood or from a hypothetical extramedullary tissue pool of granulocytes, in addition to the blood. Blood granulocytes were labeled in vitro with DFP32, infused, and their subsequent appearance in induced inflammatory exudates studied. From a comparison of the maximum blood granulocyte specific activity and the exudate granulocyte specific activity it was evident that at least three-fourths of the exudate neutrophils were derived directly from the blood. To determine if more than three-fourths of the exudate neutrophils were from the blood the proportion of granulocytes in an exudate 4 hours old which had left the blood during each of the preceding 4 hours was calculated. It was found that approximately 36% of the granulocytes in a 4 hour exudate had left the blood during the first hour of inflammation. The figures for the 2nd, 3rd and 4th hours of inflammation were, respectively, 32%, 25% and 9%, maki...

1. A fourth case of the syndrome of congenital hemolytic anemia with abnormal pigment metabolism and red cell inclusion bodies following splenectomy is described. 2. In this case an abnormality was found on paper and starch electrophoresis of the red cell hemolysate at pH 8.6. A similar abnormality has not been reported previously. 3. An increased rate of erythrocyte autohemolysis was found during in vitro incubation, Partial correction of this defect occurred in the presence of glucose or purine ribosides. 4. Genetic transmission of the defect could not be demonstrated.

SummarySingle intravenous injections of colloidal thorium dioxide in the rat produced a marked transient hypoferremia. A more persistent hypoferremia resulted from the administration of this substance daily for 3 days both in intact and in splenectomized rats. The hypoferremia producing effect of turpentine was markedly decreased by the prior administration of colloidal thorium dioxide.

The anemia of infection is associated with a number of alterations in iron metabolism. Previous studies (1-4) have shown that, with the development of inflammation, hypoferremia occurs and a decrease takes place in the concentration of the metal binding protein in the plasma. When iron is given by mouth, there is little change in the level of plasma iron and, following the intravenous injection of iron, only a transient rise in the plasma iron develops. These findings have suggested that diversion of iron from the plasma takes place in inflammation and infection. Studies in experimental animals have shown that iron is diverted primarily to the liver and spleen. Whether this iron is simply stored, since less can be used in the face of diminished hemoglobin production, or performs a special function in connection with the presence of inflammation, is unknown. These findings contrast with those associated

Summary1. Plasma copper was determined in 29 patients with sprue in relapse, 27 patients with sprue in remission, and 6 patients with megaloblastic anemia of pregnancy. 2. Mean plasma copper concentration (± 1 S.D.) in patients with sprue in relapse was 87 ± 33 μg%; in treated patients, 114 ± 33; and in patients with megaloblastic anemia of pregnancy, 159 (105 − 242). 3. It is suggested that hypocupremia in sprue may frequently be due primarily to an inability to synthesize the protein portion of ceruloplasmin. In a few patients malabsorption of copper may be a contributing factor.

Ferrokinetic studies were performed in three copper-deficient swine and the results have been compared with similar studies in 18 normal pigs. The mean value for the plasma iron turnover rate in the deficient swine was 1.76 mg./kg. day; for the red cell iron incorporation rate, 1.24 mg./kg. day; for the red cell iron turnover rate, 1.18 mg./kg. day; for the red cell life span, 13 days. Corresponding figures in the normal swine were 1.11 mg./kg. day, 1.01 mg./kg. day, 0.59 mg./kg. day and 63 days, respectively. The red cell life span was measured by the use of radioactive chromium in a total of 26 pigs. The mean erythrocyte half-life of normal cells transfused into normal pigs was 17 days. The mean half-life of erythrocytes from copperdeficient swine transfused into copper-deficient swine was 9 days. The mean half-life of red cells from control animals transfused into copper-deficient swine was 16 days while that of erythrocytes from copper-deficient swine transfused into normal pigs, was 13 days. The mean half-life of cells from iron-deficient pigs transfused into iron-deficient pigs was 19 days. It is concluded that copper deficiency anemia results from both a shortened erythrocyte survival time and limited capacity of the bone marrow to produce red cells. It is suggested that copper is an essential component of erythrocytes in swine.

SummaryDogs were given nitrogen mustard (HN2) and radioactive diisopropylfluo-rophosphate (DFP32) and the type of neutropenia which developed, whether of “early” or “late” onset, was correlated with changes observed in the blood granulocyte specific activity (BGSA) curve. The duration of phase X+II of the BGSA curve measures the time required for the storage pool of the marrow to empty by releasing cells to the blood. In dogs developing “late” onset neutropenia there was an abnormally short phase I+II while in those developing “early” onset neutropenia phase I+II was normal. These results were compared with those reported previously in vinblastine sulfate treated dogs. It was concluded that dogs with “early” onset neutropenia after HN2 must have developed a defect leading to an abnormally low rate of output cells from the marrow granulocyte reserve (MGR) to the blood. Reduced output developed despite the presence of enough cells in the MGR to maintain a normal rate of output.

1. The observations made during the administration of 114 courses of busulfan to 30 patients over a period of seven years are recorded. The responses to initial courses of therapy corresponded with those reported previously. With repeated courses of therapy, subjective improvement, the decline in the number of leukocytes, decrease of anemia and the subsidence of physical signs of the disease were as satisfactory as during the first course but tended to appear later. Patients were ambulatory and most were symptomatically and objectively improved during and after repeated courses as compared to their status beforehand. 2. Remissions were longest when the leukocyte values were 10,000 per cu. mm., or less, at the time busulfan was discontinued. Among such patients, remissions of six months or longer were seen in 85 per cent after the initial course of busulfan but in only 19 per cent after fifth or later courses. 3. Evidence of partial resistance to busulfan was seen in some cases after multiple courses of treatment. Complete resistance occurred in 15 patients upon development of the acute, myeloblastic phase of chronic myelocytic leukemia. When busulfan failed, 6-mercaptopurine and colcemide were temporarily of value; x-ray was not of benefit. After the onset of the acute phase. the median survival was three months. This mode of termination is probably no more frequent (50 per cent of cases) since the advent of busulfan therapy than before. 4. If anemia was not relieved, or a large spleen was not reduced 50 per cent in size following a course of therapy, the prognosis was poor and the acute phase imminent. 5. Thrombocytopenia in early, untreated cases was not necessarily a bad sign, nor was the use of busulfan precluded because of it. However, when thrombocytopenia appeared in a previously treated patient, without other evidence indicating busulfan toxicity, or when it occurred in a patient with three or more years of known disease, it usually presaged the development of the terminal acute phase. 6. Side effects of busulfan were significant in only one patient; this man received 8 milligrams daily, double the usual dose, for eight months and developed glossitis, anhydrosis and alopecia totalis. The major hazard in the use of the drug was the occurrence of pancytopenia. This could be related to excessive dosage. 7. Morbidity was clearly decreased. Longevity (median 42 months) was greater than in Minot’s untreated cases (median 31 months) and at least as great as that achieved by treatment with radioactive phosphorus and x-ray (median 32-41 months). Repeated courses of busulfan are considered to offer an effective and practical palliative form of therapy for chronic myclocytic leukemia, up to the time of appearance of the terminal acute myeloblastic phase.

The longevity, reckoned from the symptomatic onset of acute leukemia as well as from the initiation of treatment, of a group of 89 cases including children and adults, treated in this clinic since March, 1954, was found to exceed that of a similar group of 78 patients treated in preceding years in the same clinic. The median survival of all cases considered as a unit was increased from three to six months, as measured from the date treatment was begun. The change was found to be statistically significant as well as clinically important. The longer survival found in the 1954-57 series correlated with the greater use of three, rather than only two, chemotherapeutic agents. On analysis, improved survival was statistically proven only for individuals with lymphoblastic leukemia; for these, the median survival was 10 months after the symptomatic onset of leukemia and 7½ months after the start of treatment. Within this group, there was an even greater improvement for those whose leukocyte values immediately prior to the initiation of therapy were less than 10,000 per cu.mm.; half of these patients lived more than 12 months after treatment was begun. The reason for the exceptional longevity of patients with the lower pre-treatment leukocyte values is unknown. It appears that one can attribute at least part of the improvement to therapy. The type of statistical analysis used presents the minimum figures for improvement in median survival. When the ultimate longevity is known for the 17 per cent of the cases surviving at the closing date of the study an additional small increment may be found. The relative inefficacy of chemotherapy in myeloblastic leukemia should be a signal for greater screening of new agents in this type of tumor.

Neutrophil kinetics of acute experimental infection were studied with diisopropylfluorophosphate-32P labeling in 31 dogs inoculated intra- bronchially with pneumococci. In vitro neutrophil labeling indicated a rapid transit time through the blood in early infections, with an elevated marginal granulocyte pool sometimes preceding an elevation of the circulating granulo- cyte pool. 13 hr after infection, the circulating and total blood granulocyte pools were increased but the rate of neutrophil transit through the blood was normal. During the recovery from infection there was a marked prolongation of neutrophil blood transit time, suggesting virtually complete cessation of bone marrow release of neutrophils into the blood. Labeling of neutrophils in vivo indicated an increased rate of emptying of the bone marrow storage pool proportional to the severity of infection as measured by the fever index. The change in the blood ratio of nonsegmented to segmented neutrophils was a much more accurate index of the severity of infection than the blood granu- locyte concentration, correlating significantly with the fever index.

Summary1. Twenty-three swine were fed a diet deficient in lysine. In addition, 5 swine were given hexahomoserine (6-hydroxy-DL-norleucine), a lysine antagonist. 2. All animals developed a normocytic, normochromic anemia, which was accompanied by hypoalbuminemia and hypocupremia. No alterations from the normal were noted in the plasma iron concentration or in bone marrow morphology. The anemia responded rapidly to administration of lysine. 3. The data presented indicate that lysine is essential for normal erythropoiesis in swine.

A technic for the in vitro labeling of human platelets with DFP32 is presented, critically evaluated, and compared to in vivo methods employing DFP32 and to in vitro methods using Cr51. The initial recovery of platelets labeled in vitro with DFP32 averaged 79 per cent, but the survival curve was characterized by an irreversible initial loss of platelet radioactivity. Experiments in which platelets were simultaneously labeled in vitro with both DFP32 and Cr51 suggest that this is not due to elution of DFP32. The survival curve of platelets labeled in vivo with DFP32 shows an initial transient reduction in platelet radioactivity. It is suggested that both of these aberrations in initial survival are the result of platelet injury by DFP32. Significant "tailing" was observed in the survival curves obtained with DFP32, and possible explanations of this phenomenon are discussed. DFP32-labeled platelets circulating after 5 hours apparently survive normally and disappear from the circulation as a rectilinear function over the next 6-8 days. Although both in vitro and in vivo labeling methods employing DFP32 provide a meaningful approximation of platelet lifespan, the initial and terminal aberrations of the survival curves greatly complicate further interpretation. Dextran had no detectable effect on platelet survival, and epinephrine, Mecholyl, and cutaneous vasodilatation did not alter the platelet count or the specific activity of circulating labeling platelets in human subjects. The problem of initial platelet survival and the question of an extravascular or marginal platelet pool is discussed in the light of these data.

1. A total of 108 erythrocyte protoporphyrin determinations has been made in 66 normal individuals. The geometric mean ± standard error of the mean was 31 (26-38). 2. A total of 196 determinations of plasma iron in 92 normal individuals was made. The mean ± standard error of the mean was 104.7 ± 3.4 µg per cent. 3. In a total of 150 determinations of plasma copper in 105 normal individuals, the mean ± standard error of the mean was 118.6 ± 1.2. µg per cent. 4. No significant difference in plasma iron was noted between the sexes but in females the plasma copper was significantly higher and the erythrocyte protoporphyrin slightly higher than in males. 5. Erythrocyte protoporphyrin, plasma iron, and plasma copper determinations have been made in over 112 patients with a variety of clinical conditions associated with anemia. In general, it was found that in pernicious anemia in relapse the erythrocyte protoporphyrin values were normal, the plasma iron normal or high and the plasma copper usually normal. Anemia due to iron deficiency as well as the anemia of infection were accompanied by high values for erythrocyte protoporphyrin, hypoferremia and hypcrcupremia. In nephritis with anemia the erythrocyte protoporphyrin was generally increased, the plasma iron low or normal and the plasma copper increased. Anemia associated with lymph node disorders or leukemia was accompanied by a normal or high EP, a low or normal plasma iron and an increase in plasma copper. Thalassemia major was found to be accompanied by both hypercupremia and hyperferremia; in thalassemia minor the serum iron values were normal although hypercupremia was found. Hyperferremia was noted in aplastic anemia. In cases of plumbism the erythrocyte protoporphyrin was markedly increased. Hypocupremia was noted only twice, in one patient with severe nephritis and hypoalbuminemia and in one patient with hemochromatosis. ACKNOWLEDGMENTS For samples of plasma from patients with thalassemia we are indebted to Drs. W. N. Valentine, Rochester, N. Y., P. Sturgeon, Los Angeles, and L. K. Diamond, Boston. The following gave valuable technical assistance: Misses Mary Iles, Betty Tatting and Wanda Worth.

1.1. Total copper and ceruloplasmin concentrations have been determined in the serums of normal subjects, patients with Wilson's disease and relatives of patients with Wilson's disease. 2.2. Total serum copper was determined in 205 normal subjects. The mean value ±1 standard deviation (S.D.) was 114 ± 17.4 μg./100 ml. In two subjects the values were 68 μg./100 ml.The mean value ± 1 S.D. in thirty-six patients with Wilson's disease was 61 ± 20.8 μg./100 ml. In seven patients the values were within 2 S.D. of the normal mean. Five normal relatives of patients with Wilson's disease were found to have serum copper values below 79 μg./100 ml. 3.3. Calculation of ceruloplasmin values from the 205 normal serum copper values gave a normal range, defined as ±2 S.D., of 23 to 44 mg./100 ml. Ceruloplasmin was measured immunologically in ten normal subjects. The mean value ±1 S.D. was 34 ± 4.0 mg./100 ml. The immunologically determined values for ceruloplasmin in the two normal subjects with low serum copper values were 14 to 22 mg./100 ml. By the same method the mean value ±1 S.D. in twenty-eight patients with Wilson's disease was 9 ± 5.2 mg./100 ml. In all twenty-eight patients the value was less than 23 mg./100 ml. Ceruloplasmin values between 19 and 23 mg./100 ml. were observed in eight normal relatives of patients with Wilson's disease. 4.4. Within the group of patients studied there was poor correlation between the ceruloplasmin concentration and the duration and severity of the clinical manifestations. 5.5. It is suggested that a decreased concentration of ceruloplasmin is not the single uncomplicated determinant of the disease.

1. A deficiency of pteroylglutamic acid has been produced in 32 swine fed a purified diet containing casein and supplemented with seven B vitamins, sulfasuxidine and a folic acid antagonist. The casein was fed at two levels, 10 and 26 per cent. Two types of casein were used: a crude preparation possessing significant "extrinsic factor" activity and a purified casein with little activity. 2. The hematologic manifestations observed were (a) severe macrocytic anemia, (b) leukopenia, due to a proportionately greater reduction in polymorphonuclear than in mononuclear cells, (3) slight thrombocytopenia, and (4) hyperplastic bone marrow with an increase in immature nucleated red cells which resemble the megaloblasts seen in the bone marrow of patients with pernicious anemia. 3. The feeding of a 26 per cent rather than a 10 per cent crude casein diet did not prevent but did delay the onset of the blood changes. Anemia developed most rapidly in the animals receiving 10 per cent purified casein. 4. The group receiving 26 per cent casein developed a greater degree of macrocytosis in the same period of time than did the group receiving 10 per cent casein. In all groups the degree of macrocytosis increased as the duration of the anemia increased. 5. The hematologic manifestations were not delayed nor was their development prevented by the intramuscular administration of 15 U.S.P. units of liver extract every 15 days. 6. The blood and bone marrow returned rapidly to normal following the administration of pteroylglutamic acid, pteroyldiglutamic acid, pteroyltriglutamic and pteroylheptaglutamic acid. Thymine and xanthopterin had little or no activity. Tyrosine, adenine and uracil were inactive. 7. Purified liver extracts and crystalline vitamin B12 were found to possess some hemopoietic activity in several animals but the activity was considerably less than that of the pteroylglutamic acid compounds. 8. The urinary excretion of "tyrosyl" (hydroxphenyl compounds) was not abnormal in the pteroylglutamic acid deficient pigs and was not altered by either pteroylglutamic acid or liver extract therapy. 9. The urinary excretion of allantoin and uric acid did not differ significantly from the normal. Immediately following therapy with pteroylglutamic acid, however, in association with the reticulocytosis and lasting for the same period, there was a marked increase in the excretion of allantoin. 10. The results suggest that both pteroylglutamic acid and a factor in liver extract similar to or identical with vitamin B12 are required for normal hemopoiesis in the pig. ACKNOWLEDGEMENTS The crude methylfolic acid antagonist, xanthopterin, and the pteroylglutamic acid compounds, with the exception of pteroylheptaglutamic acid, were kindly furnished by the Lederle Laboratories, Pearl River, New York, through the courtesy of Dr. T. H. Jukes and Dr. S. M. Hardy. Sulfasuxidine was generously furnished by Sharp & Dohme, Inc., Philadelphia, Pa., through the courtesy of Dr. W. A. Feirer. Pteroylheptaglutamic acid and Natola were supplied by Parke, Davis & Company, Detroit, Mich., through the courtesy of Dr. A. E. Sharp and Dr. J. J. Pfiffner. Biotin was obtained from Hoffmann-LaRoche, Inc., Nutley, N. J., through the courtesy of Dr. E. L. Sevringhaus. The vitamins, with the exception of pteroylglutamic acid and biotin and including vitamin B12 were kindly furnished by Merck and Company, Inc., Rahway, N. J., through the courtesy of Dr. A. Gibson and the late Dr. D. F. Robertson. Experimental liver extracts (No. 1124, 1063, 1066 and 1067) were generously furnished by Armour and Company, Chicago, Illinois through the courtesy of Dr. E. E. Hays. We are indebted to Mrs. Darlene Kehl, Mr. George Trappett, and Mr. Ocie Hadley for technical assistance.

1. Thirteen cases of plasma thromboplastin component (PTC) deficiency or Christmas disease are reviewed and summarized. Of these cases, only in one were the whole blood coagulation time and prothrombin consumption normal. 2. Two cases of a mild bleeding disorder, occurring in brothers with slightly prolonged whole blood coagulation times and normal prothrombin consumption are described. 3. In these two cases, the thromboplastin generation test revealed the deficiency of a serum factor essential for normal thromboplastin generation. A mild deficiency of PTC was demonstrated by the correction of the serum deficiency by the addition to the patients’ serum of a partially purified preparation of PTC. 4. The differentiation of PTC deficiency from hemophilia is discussed. 5. Mild bleeding disorders due to a moderate reduction of antihemophilic globulin (AHG) or PTC can be differentiated by the use of the thromboplastin generation test.

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.

Normal dog plasma and serum, human, rat, and Swiss-Webster mouse plasma, phytohemagglutinin, sheep red cells, mumps and influenza vaccine, fibrinogen, and endotoxin injected before irradiation led to an increased number of endogenously derived spleen colonies in irradiated mice. Spleen weight and uptake of radioactive iron and iododeoxyuridine into such spleens were also increased. The relationship between these parameters of splenic hematopoiesis was unchanged by plasma injection suggesting that, while the number of colonies was increased, the composition of individual colonies was unchanged. This conclusion was supported by studies on plethoric mice in which splenic erythropoiesis is abolished. Increased splenic hematopoiesis was accompanied by an increase in the volume of packed red blood cells 10 days after irradiation. The total volume of plasma injected, the number of days of plasma injection preceding irradiation, and the route of administration were all important variables influencing the effect of plasma injections. Crude fractions of human albumin and gamma globulin, cortisol, C57BL (maternal) and DBA (paternal) mouse plasma, and isogeneic plasma were without effect. The ineffectiveness of isogeneic and closely related allogeneic plasma rendered unlikely the hypothesis that this effect represented the presence of homeostatic hematopoietic regulating factors in plasma. The increased hematopoiesis induced with plasma appeared to be limited to the spleen, for increased bone marrow hematopoiesis was not detected. Certain observations suggested that the effect of plasma may not be due to an antigenic or an inflammatory effect. From current observations, it was unclear whether the increased colonies induced by plasma were representative of expansion of the colony-forming cell pool or of increased efficiency of growth of the fraction surviving irradiation.

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.

The effect of a single injection of vinblastine sulfate was studied in 50 mongrel dogs. Nine of 34 dogs given 0.2 mg./Kg. of VLB died with gastrointestinal toxicity and the mortality rate increased as the dosage of VLB was increased. The morphologic pattern of leukocyte suppression and recovery in the bone marrow and blood was studied in detail in surviving animals. The cells of the bone marrow were markedly affected by VLB. Within 4 hours there was an increase in the number of cells in metaphase and, by day 1, virtually all proliferating leukocytes and erythrocytes had disappeared. An orderly repopulation of the bone marrow followed. The neutrophils, eosinophils, lymphocytes and monocytes of the blood were all markedly altered in concentration after VLB. Each type of cell first decreased to abnormally small numbers and then increased to abnormally large numbers in the blood. The curve of disappearance from and reappearance in the blood differed for each cell type. The changes in blood neutrophil number and morphology were correlated with changes in the blood neutrophil precursor cells of the marrow. The following conclusions were reached concerning the neutrophils and the assumptions implicit to these conclusions were detailed. 1. In the dog, the marrow contains enough post-mitotic granulocytes to replace those lost from the blood for at least 3 to 4 days. 2. The release of mature neutrophils from the bone marrow is a function of the rate at which blood neutrophils are lost and proceeds normally even when the marrow granulocyte reserve is partially depleted.

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.

Strauss and Brokaw1 recently reviewed and evaluated various clinical and metabolic changes in pernicious anemia. They concluded that "the occurrence of numerous manifestations of Addison's disease of the suprarenal capsule in Addisonian pernicious anemia in relapse suggest that functional adrenocortical insufficiency may be present in certain cases of this condition." It was pointed out that, in addition to certain clinical similarities between Addison's disease and pernicious anemia, there is indirect evidence of impaired adrenocortical function in pernicious anemia. Thus, patients with pernicious anemia were unable to conserve sodium chloride or excrete a water load rapidly; the oral glucose tolerance curves . . .

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.

1. Two patients with microcytic hypochromic anemia, hyperferremia, and hemosiderosis have been described .A partial hematologic remission was observed in both patients following the administration of pyridoxine. 2. Interruption of pyridoxine therapy resulted in reticulocytopenia, a decline in the concentration of hemoglobin and a decrease in free erythrocyte protoporphyrin. Increased excretion of kynurenine and kynurenic acid, but not xanthurenic acid, was observed in the urine of one of the patients following administration of tryptophane. The excretion of tryptophane metabolites was within normal limits in the second patient. 3. Reinstitution of pyridoxine therapy was followed by reticulocytosis, an increase in free erythrocyte protoporphyrin and an increase in the concentration of hemoglobin. The excretion of tryptophane metabolites in the urine following the administration of tryptophane was within normal limits in both patients. However, microcytosis, hypochromia, anemia and hyperferremia persisted. 4. The administration of brewer’s yeast, Valentine’s liver extract, calcium disodium ethylenediamine tetra-acetic acid, ethinyl estradiol, pantothenic acid, pyridoxal, pyridoxamine, ascorbic acid, niacin, riboflavin, glutamic acid and tryptophane failed to elicit a further hematologic response. 5. These patients have been compared with the seven other reported cases of "pyridoxine-responsive" anemia.

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.

SummaryIt has been found that in patients with hepatolenticular degeneration (Wilson's disease) the level of uric acid in the serum is reduced and the amount of uric acid excreted in the urine is increased. Following the intravenous administration of uric acid, such patients excrete uric acid in the urine more rapidly than do normal subjects. No correlation was observed between the excretion of copper and that of uric acid. It is suggested that the alterations in uric acid metabolism are due to an inability of the renal tubules to reabsorb uric acid efficiently.

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.

Mice were irradiated repetitively at 6 wk intervals. The proliferative capacity of the hematopoietic stem cell compartment was studied after each irradiation and compared to that of age-matched controls which had been irradiated only once. Hematopoietic proliferation capacity was measured by determining the number of spleen colonies, splenic iron uptake, spleen weight, and volume of packed red cells 10 days after irradiation. 6 wk after the first irradiation, the hematopoietic compartment was apparently supranormal in size for, when such mice were again irradiated, their postirradiation hematopoiesis was in excess of that of the controls. Thereafter, there was a steady decline in regenerative capacity with each sequential irradiation. After the sixth irradiation, the number of spleen colonies and iron uptake were reduced to one-fifth of that of singly irradiated controls. A decline in body weight and an increase in irradiation mortality accompanied the decline in postirradiation hematopoiesis. The degree of measured decline in hematopoietic proliferative ability was less than that observed by other investigators who studied the effect of serial transplantation of cells upon their ability to proliferate. Furthermore, even after the sixth irradiation, a marked stimulation of postirradiation hematopoiesis was induced by bleeding the animals before irradiation.

1. The peripheral blood changes in uncomplicated kala-azar are those of pancytopenia; namely, anemia, leukopenia and thrombocytopenia. The red blood cell morphology is normal and there is very little evidence of increased erythrogenic activity. The leukopenia is due to a reduction in all types of cells, especially neutrophils. 2. When the disease is complicated by other infections the anemia is more severe and anisocytosis, poikilocytosis, and polychromatophilia may appear and normoblasts may occasionally be seen in the peripheral blood. Leukocytosis may develop, the leukopenia may persist or the syndrome of agranulocytosis may intervene. 3. As the duration of the disease increases, the spleen tends to become larger and the anemia, leukopenia and thrombocytopenia become progressively more severe. Leukopenia generally appears first, followed by anemia and finally thrombocytopenia. The degree of leukopenia, anemia and thrombocytopenia follow closely the degree of splenic enlargement. 4. The bone marrow in kala-azar is hyperplastic and infiltrated by reticulo-endothelial cells. In spite of this there appears to be an abundance of blood forming tissue, especially erythropoietic tissue. 5. Differential cell studies on preparations of sternal marrow reveal a marked reduction in the polymorphonuclear neutrophils and eosinophils. Myeloblasts, promyelocytes, neutrophilic myelocytes and metamyelocytes are present in approximately normal proportions as are the lymphocytes and monocytes. Plasma cells are somewhat increased. Erythroid cells, especially polychromatic normoblasts are numerous and the leukocyte-erythroid ratio is altered, more than the normal proportion of normoblasts being found. Megakaryocytes are present in normal or slightly reduced numbers. Staining abnormalities are noted in these cells and there is a striking reduction in platelet production. 6. During effective anti-leishmanial therapy parasitized reticulo-endothelial cells disappear and the percentage of reticulo-endothelial cells gradually diminishes as the polymorphonuclear neutrophils increase. There is a significant increase in the eosinophilic cells. Lymphocytes become more numerous and the plasma cells diminish in number. Nucleated red cells become less numerous and the leukocyte-erythroid ratio returns to normal. At the same time the proportion of orthochromatic normoblasts to polychromatic normoblasts increases. The relative number of megakaryocytes increases and platelet formation from the megakaryocytes is accelerated even beyond the normal. Huge groups of platelets are frequently seen in the marrow smears. A rise in platelets in the peripheral blood takes place late, after there has been a significant rise in hemoglobin and leukocytes. The three cellular elements are restored to normal in the peripheral blood in the same order as their reduction from normal. 7. Evidence is presented which contradicts the view that the pancytopenia is due to a crowding out of the bone marrow by reticulo-endothelial cells. 8. Certain similarities between the hematologic changes in this disease and those accompanying the hypersplenic syndromes are noted.

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.