ALBERT

Description: By means of fractional denaturation, it is possible to follow the progress of alkaline degradation of resistant hemoglobins. A characteristic pattern was established for the fetal compound. The alkali resistant hemoglobin fractions in Mediterranean anemia and in some acquired hematologic conditions (acute leukemia and chronic aregenerative anemia) behaved like the fetal pigment. In sickle cell anemia only 4 out of 11 specimens were found to have fetal hemoglobin, whereas the others seem to have a fetal-like compound. Similarly, in 2 members of one family with hereditary spherocytosis, the resistant pigment was also fetal-like. The significance of these findings for the hypothesis that the resistant hemoglobin fractions in these disorders represent either a continuation or a reactivation of the production of the embryonic pigment is discussed.

Description: 1. In 9 patients with various types of megaloblastic anemia responding to treatment with vitamin B12, folic acid or liver extract, no significant deviations from the normal amounts of total or polymerized DNA were observed in the nuclei of red blood cells in marrow smears. 2. During the maturation of megaloblasts in the bone marrow there is a gradual loss of nuclear DNA. 3. This pattern is quantitatively and qualitatively similar for normal marrow and for that of pernicious anemia in relapse and after treatment.

Description: 1. Folic acid, when administered alone, did not prevent the development or progression of subacute combined degeneration in 12 of 22 patients receiving this agent for from twelve to twenty-five months. 2. One patient with total gastrectomy and a macrocytic anemia developed subacute combined degeneration after five months of folic acid therapy. 3. Neurologic disease did not develop in 6 pernicious anemia patients treated with folic acid and liver extract for three and one-half to thirty-nine months. 4. In 10 pernicious anemia patients with good nutrition, neurologic relapses did not progress when liver extract or vitamin B12 therapy was instituted, even though folic acid therapy was continued. In 2 patients with abnormal nutrition and complicating organic abnormalities, nervous system disease progressed after institution of liver extract therapy. 5. Our observations are best explained by the theory that the hematologic and neurologic manifestations of pernicious anemia and other macrocytic anemias associated with gastro-intestinal tract pathology and inadequate nutrition are due to a deficiency of more than one substance. The administration of folic acid may improve the hematologic status but induce a deficiency of another substance or substances, e.g., vitamin B12, which are essential for the maintenance of a normal blood picture and the integrity of the central nervous system. This deficiency will eventually result in the development of a suboptimal blood picture or subacute combined degeneration of the spinal cord, or both. 6. The hematologic status of patients with pernicious anemia is not maintained in a more satisfactory state by supplementation of liver extract or vitamin B12 therapy with folic acid. 7. Folic acid therapy did not produce neurologic disease in patients with iron deficiency anemia who had free gastric hydrochloric acid in their gastric secretions and presumably sufficient intrinsic factor. It did not influence response to ferrous sulfate therapy. 8. Patients with sprue, nutritional macrocytic anemia and other macrocytic anemias associated with gastro-intestinal tract pathology who are treated with folic acid should also be given supplemental liver extract or vitamin B12 to insure against the development of nervous system disease.

Description: 1. Normal gastric juice (intrinsic factor) and vitamin B12 together form a thermolabile hemopoietic factor which ripens megaloblasts in vitro, both gastric juice and B12 alone being inactive. 2. The hemopoietic factor in normal serum which ripens megaloblasts in vitro also appears to be thermolabile, heating to 56 C. for 2 hours destroying some of its activity. 3. The relationship of these factors is discussed and an extra-gastric as well as a gastric source of intrinsic factor is postulated.

Description: Differential Diagnosis. An unresponsive anemia associated with a leukoerythroblastic peripheral blood picture may be seen in such varying clinical conditions as carcinomatosis, leukemia in early phase, Hodgkin’s disease, toxic aplastic anemia, hemolytic anemia, myelosclerosis, myeloma, syphilis, tuberculosis, lipoidosis, Leishmaniasis and infective hepatitis. From the clinical course of the disease and routine methods of investigation, the majority of these diseases could be excluded. It was soon apparent from the appearance of the marrow and the result of the marrow culture that this disease was to be regarded as a primary blood dyscrasia. There is a group of such dyscrasias which is characterized by abnormalities in both erythro- and leukopoiesis. All gradations between erythremic myelosis (Copelli,2 Guglielmo,3 Leitner et al.6) and myeloblastic leukemia may occur. The case here described is neither one of erythremic myelosis nor one of acute leukemia. Heilmeyer and Schöner5 describe a true chronic erythroblastosis of adults which, however, is accompanied by a leukocytosis, never present in this case. Several transitional cases, where there is neither a predominant erythroblastic nor a proponderating myeloblastic hyperplasia, have been reported. In these there is, however, evidence of a neoplastic hyperplasia of both erythroid and myeloid tissue. Such cases have been described by Penati,10 Moeschlin,7 Moeschlin and Rohr,8 Rohr,11 Harvier et al.,4 and Stahel.12 The course of six months in the present case resembles that of Moeschlin.7 It is of academic interest to differentiate between a true aplastic anemia, an aplastic initial phase of leukemia and the mixed neoplastic forms of the erythroleukemic group. Etythroblasts in cases of this group frequenfly show "megaloid" changes (as do those in acute leukemia). A series of transitional forms from normoblasts to megaloblasts through "megaloid" forms may be seen. This could be due to a relative deficiency in nutritive factors consequent on the abnormal cellular proliferation. Similarly in many types of acute leukemia this type of cell, when cultured in normal serum, reverts to the normal maturation rate and morphology of the normoblast. So do the cells of aplastic anemia, where the ultimate defect is further removed. When, however, an abnormal normoblastic maturation rate is found, a qualitative, probably neoplastic, change in the nature of these cells is suggested, since the same changes may be found in the myeloid series in cases of myeloid leukemia. Hence marrow culture may prove of diagnostic value in differentiating these somewhat allied conditions. This case is one of progressive, irresponsive anemia with a peripheral leukoerythroblastosis, and bone marrow showing a considerably lowered leukoerythrogenetic ratio with a relative myeloid predominance of early forms and of atypical blast cells (paramyeloblasts). Culture of the bone marrow established that the rate of maturation of both the erythroid and myeloid tissue was abnormal, although the defect was more marked in the erythroid series. On these findings a diagnosis of erythroleukemia seems justified. Since this report, 2 similar cases investigated by marrow culture have shown comparable maturation abnormalities.

Description: 1. A simple method for distinguishing between prothrombin and prothrombin conversion accelerators has been described. 2. A deficiency in prothrombin conversion accelerators in the early days after major surgical operations has been observed.

Description: 1. Seventy patients with acute leukemia were treated with various folic acid antagonists. Sixty-five survived for a sufficient length of time to evaluate the effect of the therapeutic agents. Types of leukemia observed included 38 cases of acute lymphatic leukemia, 23 patients with acute monocytic leukemia and 4 with the acute myelogenous type of leukemia. 2. The best results, both clinically and hematologically, were obtained in acute lymphatic leukemia. Although the most satisfactory results were observed in the youngest age group, excellent remissions were produced in older individuals. Of 65 cases observed, an excellent clinical and hematologic remission was observed in 11 patients, a partial emission in 19 subjects, and no response in 35 individuals. 3. When a response occurred, a rater definite hematologic pattern was noted. An increased platelet count in most instances was the first evidence of regeneration and occurred in about the third or fourth week. The neutrophilic polymorphonuclear leukocytes began to regenerate at about the same time and an increase in their number was followed by a rise in the erythrocyte count. 4. Observations of the bone marrow indicated that although excellent clinical and hematologic remissions might occur, primitive leukemic cells were still present. In some instances megaloblasts were observed in addition to a peripheral macrocytosis and anisocytosis of erythrocytes. 5. Toxic manifestations were common. These included glossitis, ulceration of the oral cavity, nausea, vomiting, diarrhea and alopecia. In one instance there was ulceration of the entire gastro-intestinal tract, including the esophagus and colon. Hematologic toxic reactions included thrombocytopenia, leukopenia and anemia. Aplasia of marrow tissue was observed in 1 instance. In many instances the margin of safety between a toxic reaction and death was indeed small. 6. It should be emphasized that in no instance has a cure of leukemia resulted from treatment with a folic acid antagonist although prolonged remissions have occurred.

Description: 1. The concentration of acid soluble phosphorus, lipid phosphorus and nucleic acid phosphorus was determined in the red cells and polymorphonuclear leukocytes of the rabbit. The concentration of acid soluble phosphorus was of the same order in both types of cells, but there was more lipid phosphorus and nucleic acid phosphorus in the polymorphonuclear leukocytes than in the red cell. Over 80 per cent of the nucleic acid in white cells was of the desoxypentose type. 2. Immature cells were produced in the peripheral blood stream of rabbits either by the administration of phenylhydrazine or by successive bleedings. Either treatment caused a fall in the concentration of hemoglobin, the red cell count and the hematocrit with an increase in the mean corpuscular hemoglobin. There was also an increase in the concentration of acid soluble phosphorus and lipid phosphorus and a great increase in the concentration of nucleic acid phosphorus which was coincident with the increase in number of immature cells as judged by the reticulocyte count. Almost all of the excess nucleic acid was of the pentose type. 3. The basophilia of the immature cells disappeared after treatment with ribonuclease or after hydrolysis with normal hydrochloric acid. Desoxyribonuclease was without effect. 4. These results suggest that the basophilic substance of the immature rabbit red cell, and presumably the reticulum of the reticulocyte, is of cytoplasmic rather than nuclear origin.

Description: In 2 cases of chronic hemolytic anemia with erythrocyte fragility to cold and acid, although the abnormality resides in the red blood cells, serum, whether from the patients or from normal controls, is necessary for the hemolysis. Complement is most likely not responsible for the serum hemolytic activity, because the latter, while similar to complement in certain properties, may be completely dissociated from it. From human serum the euglobulin separated by CO2 and sodium sulphate and the pseudoglobulin I obtained by sodium sulphate precipitation possess hemolytic activity for the cells of the patient and not those of the control. The hemolytic activity of these globulin fractions is increased by chilling and by addition of acid. It is inhibited by serum pseudoglobulin II and albumin. Guinea pig serum aids human serum in its hemolytic activity on susceptible human cells. This augmenting effect of the guinea pig serum is not due to its complement content, nor due to its hetero-hemolysin for all human erythrocyte. In view of the data presented in this and the preceding papers, a new syndrome is suggested which can be clearly differentiated from paroxysmal cold hemoglobinuria and from paroxysmal nocturnal hemoglobinuria.

Description: Experiments are described which attempt to define the relation of methemoglobin to hemolysis. A comparison was made in vitro of the resistance of erythrocytes containing chiefly oxyhemoglobin or methemoglobin to hemolysis by saponin, hypotonic saline, bile salts and lysolecithin. These data are interpreted to indicate that intracorpuscular methemoglobin does not alter the resistance of erythrocytes to hemolysis by these lysins, although there are certain peculiarities in the results with bile salts and lysolecithin. In the case of bile salts and lysolecithin, oxyhemoglobin or methemoglobin may have an equal or different inhibitory effect on hemolysis depending upon the conditions of pH and temperature. Chronic methemoglobinemia in dogs maintained by the administration of sodium nitrite does not produce an anemia. These data suggest that the formation of methemoglobin is an independent effect and not a part of the hemolytic action of certain chemical agents.

Description: Quantitative changes in two serum anti-proteases in several types of adult leukemia are described. High anti-chymotrypsin and low anti-rennin concentrations were observed in the acute leukemias while the reverse was found in chronic forms of the disease. The relationship between the enzyme inhibitors and disease activity in leukemia is similar to that reported in cancer patients.

Description: 1. A series of 212 cases of acute leukemia classified according sex and age is presented. Fifty-eight out of 212 were diagnosed by means of phase contrast microscopy. 2. In addition to this personal series, other series collected from the literature tile reviewed. These include both acute and chronic leukemia. 3. The thorough analysis of maximal frequency of the disease according to age, shows that there is, in the course of life, an inverse relation between lymphoid and myeloid leukemias as to the period of life in which they appear. This relation favors the view that a particular ratio of equilibrium must exist between the two systems: lymphoid and myeloid. 4. It is suggested that such an equilibrium might depend upon endocrine factors.

Description: 1. Five human lymphatic and myeloid reticulosarcomas were transmitted to fowls in series. 2. The pathologic changes produced in fowls consisted in proliferation of reticulo-endothelial cells (reticulosarcoma and allied conditions) or those of immature myeloid cells (myelosarcoma, myelosis or myeloid metaplasia). These different forms of leukosis may occur alternately in subsequent generations or simultaneously in the same bird. Neither erythroleukosis nor lymphatic leukosis was encountered. These observations conform with our earlier experience gained in experimental studies of human leukemia. 3. The blood picture of infected fowls was consistently aleukemic either with marked leukopenia or slight leukocytosis. There was usually an increase in mature granulocytes at the expense of lymphocytes. Anemia was usually slight. 4. The causative agent can pass through a Chamberland L2 filter. 5. The infective agent can be demonstrated in the blood of patients, in fluid from the thoratic cavity, in the enlarged lymph nodes as well as in the human tumors.

Description: A case of cholelithiasis is reported in a child with the severe form of Mediterranean anemia. The patient was one of a group of children with this disease but without biliary tract symptoms on whom a roentgenographic survey was carried out. Cholecystitis subsequently occurred in this patient, and a cholecystectomy was performed. Cholelithiasis occurs as a complication of severe Mediterranean anemia, less frequently than in spherocytic or in sickle cell anemia but on a comparable etiologic basis.

Description: MOVITT, E. R.: Megaloblastic erythropoiesis in patients with cirrhosis of the liver Blood 5: 468-477 (May), 1950. In figure 2, pages 471, parts (a) and (b) were reversed. The upper photomicrograph (a) actually shows bone marrow after treatment; the lower (b) shows marrow before treatment.

Description: "L. E." cells and rosettes were demonstrated in the blood of patients with systemic lupus erythematosus without the use of anticoagulants. Concentration technics using defibrinated blood and unmodified blood in silicone coated tubes were employed.

Description: 1. Porphyrin and hemoglobin metabolism have been studied in a case of photosensitive or congenital porphyria with the aid of glycine containing N15. The case was that of a 4 year old girl who also exhibited splenomegaly and hemolytic anemia. The reticulocytes and fecal urobilinogen were markedly increased and the bone marrow was hyperplastic-normoblastic. The urine contained large amounts of uroporphyrin I and lesser amounts of coproporphyrin I, while the reverse was true in the feces. Splenectomy resulted in disappearance of anemia, together with marked reduction of porphyrin excretion. 2. In two separate periods of study a rapid incorporation of glycine N15 was observed in the hemoglobin protoporphyrin, in both coproporphyrin isomers of the urine and in the type I isomer in the feces; also in the uroporphyrin I of the urine, and in the protoporphyrin and stercobilin of the feces. 3. The N15 data obtained for the urinary and fecal coproporphyrin and the urinary uroporphyrin, clearly reveal a synthesis similar in character to that of the hemoglobin protoporphyrin. The striking decline in formation of copro- and uroporphyrin after splenectomy is believed to be correlated directly with the reduced erythropoiesis incident to removal of the excessive hemolysis. 4. In both experiments the peak concentration of N15 was earlier and greater in the copro- than in the uroporphyrin. This, together with the closely similar slopes of decrease of the N15 curves for the two substances, does not appear to support a transition of uro- to coproporphyrin in the present case. The data are more consistent with the view that coproporphyrin I is a precursor of uroporphyrin I, or that both porphyrins are synthesized independently from common precursors. 5. The amount of coproporphyrin III in the excreta was too small to permit N15 analysis except in one period early in the first experiment; here the value was approximately the same as that of the copro- I for the same period, indicating a closely related synthesis.

Description: 1. Erythrocyte and plasma cholinesterase activities of the blood were determined by Michel’s potentiometric method in 20 healthy individuals and in 20 patients ill with various blood dyscrasias. 2. The mean erythrocyte cholinesterase activity in the blood of healthy individuals was 0.71 ± 0.11 Δ pH/hr. and the mean plasma cholinesterase activity was 0.98 ± 0.17 Δ pH/hr. 3. Low erythrocyte cholinesterase activity was found in the blood of patients ill with pernicious anemia or ill with hypoplastic anemia. 4. High erythrocyte cholinesterase activity was found in the blood of patients suffering with anemia secondary to hemorrhage and of patients ill with pernicious anemia or macrocytic anemia of nontropical sprue in early remission. 5. The plasma cholinesterase activity of blood from patients ill with blood dyscrasias usually reflected the general clinical status of the patients and, in general, was related to the albumin concentration in the serum. 6. The low erythrocyte cholinesterase activity increased following multiple blood transfusions and during a reticulocyte response in a case of hypoplastic anemia, whereas the plasma cholinesterase activity remained unchanged. 7. The erythrocyte cholinesterase activity in patients ill with pernicious anemia or macrocytic anemia of nontropical sprue increased rapidly during therapy beginning with the reticulocyte response and continuing during the period of increased erythropoietic activity. The plasma cholinesterase activity increased slowly parallel to improvement in hematologic and clinical status. 8. The possible relationships between alterations in erythrocyte cholinesterase activity in blood dyscrasias and reticulocytosis and erythropoiesis are discussed.

Description: 1. A general description is given of the known effects of a series of allelic genes, W, Wv, w,, which are responsible for varying degrees of a macrocytic anemia as well as for differences in viability, gonad development and intensity and location of hair pigment. A summary is given of previous investigations into the basic nature of these anemias. 2. The mean erythrocyte count for each genotype is given for sixteen day embryos, and for zero day, two weeks and four weeks after birth. All determinations are based on comparisons of normal and anemic littermates. 3. A detailed description is given of the methods used to obtain blood samples from embryos. In all three severely affected genotypes the anemia is already well established at the sixteenth day of embryonic life. 4. In each interval thereafter, some increase in erythrocyte level is found in all genotypes. All genotypes, normal and anemic, made approximately the same relative increase during each time interval, indicating the same relative response to the physiologic stimulus of the individual’s growth. 5. Certain genotypes are lethal at particular stages. It is suggested that these deaths are due to a failure of blood formation to reach the absolute level necessary for continued development and resistance to minor noxious stimuli at these critical periods of rapid growth. 6. Certain other factors possibly affecting blood formation or survival were tested. The presence, absence or degree of anemia of the mother were found to have no effect on the erythrocyte level of newborn young of the slightly anemic Wvw genotype. Continuing injections of diluted whole blood into extremely anemic WWs led to survival for more than 100 days. Two WW individuals survived spontaneously to seven months, indicating that there are influences, as yet unanalyzed, which can lift even this genotype to a level where it can produce sufficient erythrocytes to maintain itself for a considerable period.

Description: The Rh antibody titer was studied during pregnancy and after delivery in 11 cases in which previously sensitized Rh-negative mothers had Rh-negative babies. In addition, 1 case was studied in which a type Rh1 mother, previously sensitized to the rh'' factor, gave birth to a type Rh1 baby. In all cases the babies were normal at birth and the baby’s serum contained univalent Rh antibodies equal in titer to those present in the maternal serum. In no case was there a rise in the antibody titer in the maternal serum either during the pregnancy or after the baby was born. Thus, it has not been possible to confirm claims concerning a "nonspecific anamnestic rise" in Rh antibody titer said to be caused by an Rh-negative baby. Two illustrative cases are described in detail. In one of the cases described, there was no appreciable change in Rh antibody titer during the pregnancy despite the use of so-called Rh hapten; nor had there been an appreciable change in titer for as long as two and one-half years previously. The Rh antibody titer of the baby’s blood dropped about 98 per cent by the end of the sixth month, while during the same period of time the maternal antibody titer fell only approximately 35 per cent. Based on the course of the Rh antibody titer of the baby’s serum, the half-life of the antibody globulin molecule is estimated to be about thirty days.

Description: Through clinical, genetic and physicochemical studies a new inherited abnormality of hemoglobin has been recognized in American Negroes. The new hemoglobin, provisionally called hemoglobin III, can be separated from both normal and sickle cell hemoglobin by electrophoretic analysis. The structural anomaly of the hemoglobin molecule is determined by a gene inherited as a simple Mendelian dominant. A distinct hemolytic syndrome which is intermediate between the benign sickle cell trait and sickle cell anemia results from the combination of the new hemoglobin with sickle cell hemoglobin. A tentative characterization of the syndrome is presented. In contrast to classical sickle cell anemia this form of sickle cell disease is characterized by a mild hemolytic anemia with slowly progressive splenomegaly in the absence of cardiac or musculo-skeletal manifestations. In vitro, the erythrocytes sickle like those of sickle cell anemia. The bone marrow shows erythroid hyperplasia, fecal urobilinogen excretion is increased, and the survival time of the erythrocytes in normal recipients is shortened, but in the patients the red cell count and hemoglobin concentration are only slightly depressed. Reticulocytosis is slight and icterus is not observed. Whereas in sickle cell anemia both parents are expected to have the sickle trait, only one parent of these individuals shows sickling, while the nonsickling parent is a carrier of the hemoglobin III. The new syndrome does not appear to be identical with that resulting from the simultaneous presence of the sickling gene and the thalassemiagene. The presence of hemoglobin III when combined with structurally normal hemoglobin is expressed as an asymptomatic carrier state. The erythrocytes do not sickle but have a high incidence of target cell deformity and increased resistance to hypotonic saline. Although there is no evidence of hemolysis in such individuals their erythrocytes are eliminated with abnormal rapidity from the circulation of normal recipients. The homozygous state with respect to this new hemoglobin has not yet been identified but may well be some already recognized atypical form of chronic hemolytic anemia. Studies are now in progress to determine the incidence of this new molecular abnormality of hemoglobin.

Description: Four new cases of congenital leukemia reported here may be added to the 14 acceptable cases in the literature. In all except 1 of these cases cells of the granulocytic series were involved; males predominated over females in a ratio of more than 2 to 1. Congenital anomalies were present in 3 of the 4 cases we have reported, indicating a growth disturbance early in fetal life. Contrary to the usual finding in childhood leukemia, cells of the granulocytic series in the 4 cases reported here exhibited a fair degree of differentiation and in this respect are similar to those which characterize the chronic form of the disease. It seems likely, therefore, that the leukemia existed during a significant portion of intrauterine life.

Description: A number of investigations were made in a hemophiliac who had become refractory to treatment with whole blood, plasma and plasma fractions. This refractoriness was traced to the presence of a circulating anticoagulant. By fractionation of the plasma it was determined that the anticoagulant was associated with the pseudoglobulin fraction of the plasma, and acted by inhibiting the action of antihemophilic globulin (coagulation globulin) present in natural blood. The antihemophilic globulin, however, was not physically altered by the anticoagulant and could be reprecipitated without loss of its antihemophilic properties. The anticoagulant did not inhibit prothrombin, thromboplastin, or fibrinogen. It was unrelated to the lipid antithromboplastin or to heparin and did not appear to be an antibody.

Description: The factors which normally limit the life of the red cell are described as being a continuous metabolic process involving the enzymatic oxidation of Hb and a terminal event, which may be hemolysis, fragmentation or phagocytosis. A number of lytic substances, such as soaps, lipids and lysolecithin-like substances can be extracted from plasma and from tissues. These substances are associated with inhibitors and accelerators to form complexes. The activity of these naturally occurring hemolytic complexes tends to be small, although it can increase to such an extent that appreciable red cell destruction results. Instances are given in which hemolysis in vivo results from the concentration of one of these lysins increasing (as when fat is fed), from an accelerator of one of the lysins being introduced (usually as a drug), and from the concentration of inhibitory material being reduced (as by a low cholesterol diet). Most hemolytic episodes are due to the establishment of a new hemolytic mechanism, which may appear after the introduction of a drug, of an agglutinin such as silicic acid or ricin, of an immune agglutinin which is not itself a hemolysin or of an agglutinin which is a lysin in the presence of complement. The various mechanisms which may result in hemolysis are discussed. The case in which the agglutinin becomes a lysin in the presence of complement presents no difficulty; in other cases the mechanism of hemolysis is not so clear, nor is it clear whether red cell destruction depends primarily on hemolysis or primarily on phagocytosis. Special processes are involved in the destruction of red cells which have intrinsic defects of structure. The abnormally thick red cells of congenital hemolytic icterus are selectively sequestered in the spleen, where there are a number of hemolytic mechanisms which can destroy them the more readily because of their abnormal shape. The sickle cell, with its poor osmotic properties, its reduced mechanical fragility, and its tendency to lose part of its structure as filaments at each disk-sickle transformation, is destroyed by processes which are probably hemolytic but less easy to specify. The flat red cells of Mediterranean disease are abnormally prone to fragmentation. In all these diseases the abnormal shape of the red cell seems to be accompanied by peculiarities in its contained hemoglobin, an observation which requires further study before its significance is clear.

Description: 1. Data have been presented, showing that it is possible to standardize conditions under which absolute eosinophil levels in the tail blood of mature male mice characteristic for different strains can be obtained. 2. The highest eosinophil levels were found in the strain of mice with the lowest susceptibility to mammary cancer among the stocks tested. 3. The absence of the mouse mammary cancer milk agent did not affect the characteristic eosinophil morning level in A or Z mice. 4. Emphasis is placed upon the necessity of rigid control of the circumstances of observation if reproduceable results are to be obtained in the study of eosinophil levels.

Description: 1. The blood cells of trout and roach consist of nucleated erythrocytes and reticulocytes, nucleated thrombocytes, coarse and fine granulocytes, and lymphocytes of varying sizes. It is difficult to distinguish any cells showing characteristics similar to those of the monocytes of mammals. Immature cells occur more frequently in the blood than is the case in mammals. 2. The coarse granulocytes very commonly escape from the blood vessels, and have been observed in large numbers in the intestinal mucosa and submucosa, in the gill epithelia, and in the peritoneum. These cells migrate to the epithelial surface, where they undergo changes in structure leading to the formation of a characteristic discharge pattern of their granules. It is proposed that the "granules" are in reality vesicles with fluid contents, which are ultimately discharged at epithelial surfaces. 3. The hematopoietic organs of these fishes are chiefly the intertubular tissues of the kidneys; in the trout the spleen is also active; in the roach, only the kidney is active; in the perch, only the spleen is active. 4. Two alternative hypotheses of blood cell formation are proposed. On the first hypothesis, the common stem cell is described as a "large lymphoid hemoblast," which gives rise to granulocytes by direct transformation and undergoes mitotic division to give rise to "small lymphoid hemoblasts." From the latter develop the erythrocytes, thrombocytes and blood lymphocytes. On the second hypothesis, the large lymphoid hemoblast, derived by transformation of reticular cells is the precursor solely of the granulocytes, and the small lymphoid hemoblast is to be derived from endothelial cells and is the precursor of erythrocytes and thrombocytes. In this case the large cell is to be compared with the "primitive white cell" of Doan, Cunningham and Sabin, and the small cell with the "megaloblast" of the same authors. No evidence however is available of the derivation of small hemoblasts from endothelial cell components of the reticulo-endothelium. 5. In the maturation of the erythrocyte in teleost fishes, there is a progressive increase in the size of the cell; in mammals and birds there is a decrease in size. In both cases there is a decrease in size in granulocyte maturation. 6. There are no essential differences between the blood cells and hematopoietic processes of the freshwater and marine teleost fishes examined.

Description: Two cases of hypoprothrombinemia of unknown cause are presented. In each case there was a marked hemorrhagic diathesis which, in Case 1, led to death. In each there were additional findings (clinical or necropsy) which suggested that the prothrombin deficiency was only one manifestation of a generalized disease. The pertinent literature is reviewed and a classification of the previously reported cases of "idiopathic hypoprothrombinemia" into "congenital" and "acquired" types is offered. Both of our cases fall into the "acquired" group. The treatment of the condition is discussed and it is pointed out that vitamin K1 oxide in large doses restored the prothrombin time to normal after the apparent failure of large doses of water soluble menadione. A rationale for this difference in response is suggested.

Description: 1. The manner of inheritance of the sickling phenomenon has been investigated in 75 kindreds, with hematologic observations on 465 persons. 2. Data are presented to support the contention that with a possible rare exception, the differential diagnosis between the sickle cell trait and sickle cell anemia is readily made. 3. The hematologic findings pertinent to a decision as to the mode of heredity of the sickle cell trait and sickle cell disease are as follows: a. Out of a total of 94 parents of children with sickle cell disease, 93 have shown the sickle cell trait. b. The incidence of sickle cell disease in segregating sibships would appear to lie between 0.18 and 0.26. c. In sibships segregating for sickle cell disease, the ratio of sickle cell trait to normal is approximately 2:1, but in sibships resulting from the marriage of a normal person with an individual with the sickle cell trait, the ratio of sickle cell trait to normal is significantly less than 1:1. d. Of the 4 children in this series who have a parent with sickle cell disease, all have the sickle cell trait. 4. The most reasonable hypothesis which will render the above data intelligible is that the sickling phenomenon is due to a gene which in single dose (heterozygous condition) produces only the sickle cell trait, and in double dose (homozygous condition), sickle cell disease. The chief and only discrepancy between theory and fact at the present time appears to lie in a deficiency of individuals with the sickle cell trait in certain types of matings. 5. No clear cut differences between normal and sickle cell trait were observed with respect to erythrocyte number, hemoglobin level, hematocrit, the cell constants, the leukocyte number or the differential count. 6. Certain minor hematologic abnormalities encountered in the kindreds under study and not obviously related to the sickling phenomenon are described.

Description: In rats, the intravascular injection of large numbers of lymphocytes obtained from the lymphatic system fails to increase the number of circulating lymphocytes. This is the result, chiefly, of a specific selective action of the lungs in removing the cells, after either intravenous or intra-arterial injection. The spleen, and perhaps the liver and kidneys also play a part in removing the transfused lymphocytes, but are not as active as the lungs in this respect. These results are similar to those obtained with the transfusion of granulocytes obtained from the peritoneal cavity of rabbits and raise the possibility that the lungs may act as a homeostatic organ in controlling the level of the white blood count. Under the conditions of this experiment, relatively large doses of ACTH and cortisone had no significant immediate effect on transfused lymphocytes. These results indicate that inadequate preservation of leukocytes in stored blood does not account for their failure to appear in the circulation after transfusion, and that better preservation will not, in all probability, alter the situation.

Description: Prothrombin and fibrinogen represent the fundamental materials in the formation of the solid clot. During the process of blood coagulation, prothrombin is converted to thrombin, and this enzyme, in turn, clots fibrinogen. The formation of thrombin is the essential step in the clotting of blood. Besides prothrombin, at least five other agents participate in the reaction. They may be divided into: a) conversion factors, agents actively participating and indispensable for the formation of thrombin, at least for physiologic needs; and b) accelerators, which increase the speed of the conversion of prothrombin to thrombin and the yield of this enzyme once the process has been initiated. The first group includes thromboplastin, calcium and a plasma prothrombin conversion factor (PPCF); the second, a serum accelerator and a platelet accelerator. Platelets may also supply a factor which accelerates the formation of fibrin from fibrinogen. Thrombin, once formed, is able to accelerate its own production (autocatalysis), thus determining an "explosive" formation of fibrin. This mechanism is of great importance in assuring efficient hemostasis. Adsorption of the thrombin on the fibrin clot and neutralization of the enzyme by the natural plasma antithrombin when it is slowly released during the retraction of the clot are two known mechanisms by which the organism protects itself from any undue extension of the intravascular thrombus.

Description: Estimates of bone marrow cellularity based upon gross appearances and upon total nucleated counts of the aspirates were compared in 50 cases. These comparisons revealed some correlation between the gross appearances and the averages of the total nucleated counts. That this correlation did not always exist in individual cases was demonstrated by the wide range in total nucleated counts and by incorrect gross appearance estimates in two cases of acute leukemia. Differential counts as obtained from particle smears and random sample smears of the same aspirates were compared in 50 cases. Such comparisons revealed certain consistent differences in cellular elements. These differences and their significance were discussed. The dilution of aspirate fluid and aspirate particles with peripheral blood was determined in 10 cases. Appreciable and unpredictble dilution of aspirate fluid was demonstrated, but none was demonstrated in aspirate particles. This dilution of aspirate fluid with peripheral blood is implicated as the cause of the inaccurate estimations of bone marrow cellularity based upon gross appearances and total nucleated counts. It is also suggested as one of the factors underlying the observed differences in the cellular elements of particle smear and random sample smear differentials.

Description: 1. The clinical features, laboratory findings and special studies of a case of photosensitive (congenital) porphyria in a 4 year old girl have been presented. This case was of particular interest in view of severe hemolytic anemia with hepatosplenomegaly. 2. Copro- and uroporphyrin I were isolated from the urine and feces. The ratio of these porphyrins in the urine varied from 1:10, to 1:30 respectively, while in the feces the ratio was reversed at about 70:1. Coproporphyrin III was isolated in much smaller amount than the type I isomer, from both urine and feces. Isomer analyses of the coproporphyrins in the excreta indicated that approximately 98 per cent was type I. 3. Prior to splenectomy copro- and uroporphyrin I were isolated in crystalline form for the first time from circulating human erythrocytes. Coproporphyrin III was also isolated in lesser amount. Uroporphyrin I was crystallized from the plasma, which also contained coproporphyrin I. Microfluorospectrometry of the bone marrow revealed large amounts of porphyrin in the developing red cells. The porphyrin fluorescence spectra indicated that at least three porphyrins were present. 4. Splenectomy was followed by disappearance of uro- and coproporphyrins from the erythrocytes, and a marked decrease in plasma, urine and feces without any essential change in the type of porphyrins excreted. The metabolic defect porphyria, was still present, but now latent in character. Reduction in porphyrin excretion was apparently related to elimination of hypersplenic hemolysis and compensatory increase of erythropoiesis. Anemia and dermal photosensitivity to sunlight disappeared simultaneously with the reduction in porphyrin excretion. 5. Efforts to reproduce the skin lesions by artificial light were unsuccessful.

Description: 1. The hemoglobin concentration in the cord blood of infants affected with hemolytic disease is very closely related to their chance of survival. If the hemoglobin concentration is plotted against the chance of survival, a sigmoid curve is obtained. Probit analysis can be applied and the results of treatment can then be summarized in terms of the severity of the cases treated. For example, in the present series the cord hemoglobin concentration corresponding to a 50 per cent chance of survival was 8.09 Gm./100 ml. 2. Application of probit analysis makes it possible to compare results obtained in different centers, for due allowance is made for any difference in severity in the cases included in the different series. 3. In the present series, despite uniform treatment (exchange transfusion), over-all mortality fell from 37 percent in 1947 and 1948 to 3 per cent in 1950. It is shown that this can be entirely explained by a decrease in the number of severely affected infants. 4. Predictions of survival based on the cord hemoglobin concentration and cord bilirubin concentration combined are not significantly better than predictions based on cord hemoglobin concentration alone. 5. Infants with positive direct Coombs tests whose cord hemoglobin concentrations exceed 15.5 Gm./100 ml. have some risk (approximately 1 in 12) of developing kernicterus, if untreated.

Description: 1. By simultaneously sampling venous and arterial blood by cardiac catheterization or vessel cannulation, the number of leukocytes entering and leaving the lungs was observed in 12 patients on 14 occasions. 2. The intravenous administration of histamine phosphate in doses of 0.1 to 0.3 mg. (as base) over 10 to 60 seconds, was accompanied by a prompt decrease in leukocyte number in the arterial blood 20 to 60 seconds before the venous white cell count fell. This was interpreted as demonstrating that the leukocytes were removed from tine peripheral blood in the pulmonary circulation. The granulocytic series appeared to be more involved in the leukopenia, although a similar but less apparent change was noted in the agranulocytes. 3. The leukopenia persisted for 40 to 180 seconds following which the arterial leukocyte count exceeded that in the venous blood indicating a return of leukocytes from the lungs into the peripheral circulation. 4. The intravenous administration of histamine also resulted in an immediate decrease in clotting time as determined both by glass and siliconed tube technics. 5. The intravenous injection of histamine affords a relatively simple technic to study one type of leukocyte removal mechanism present in the pulmonary circulation.

Description: 1. Further studies contrasting the effect of varying doses of 4-amino-N10-methyl-PGA on a resistant variant (Ak4R) of transplanted mouse leukemia Ak4 have been reported. 2. The usual therapeutic dosage of 4-amino-N10-methyl-PGA prevented the increase in total leukocytes, delayed the visceral infiltration, and increased survival time in the Ak4 strain but was without effect in strain Ak4R. 3. In the Ak4 leukemic mice with high leukocyte counts a marked drop within twenty-four hours could be demonstrated after a single dose of 3 mg./Kg. of this drug. With the Ak4R strain there was sufficient resistance so that a dose of at least 25 mg. Kg. was needed to show the marked leukopenia in twenty-four hours. 4. The Ak4R strain was as sensitive as the Ak4 to the leukotoxic effects of 2,6-diaminopurine. This drug also was effective in prolonging the survival time of mice with Ak4R leukemia.

Description: 1. Activity-pS curves were determined experimentally for the cholinesterase of the erythrocytes in various human anemias. 2. The data were analyzed to give experimental values for the apparent dissociation constants, K8 of the active enzyme-substrate complex (ES) and K2 of the inactive complex (ES2), and the theoretic limiting velocity V. 3. The analysis showed that the enzyme in the cells of anemia is the same as that in normal cells. The abnormally high enzymic activity (initial velocity at a single substrate concentration) known to occur in various anemias and the change from low activity in pernicious anemia in relapse to high activity during treatment are attributable to changes in concentration of the enzyme. 4. The enzyme in the red cells of pernicious anemia in relapse was not activated by folic acid or liver extract. 5. It was shown experimentally that the reticulocytes and young cells of normal blood and of pernicious anemia during treatment contain much higher concentrations of enzyme than old cells. The increased concentration persists for some time after the reticulum has disappeared. 6. The relative concentration of enzyme in the red cells is a more sensitive indicator of hyperactive hematopoiesis than the reticulocyte count. 7. Evidence is presented that failure of elevation of enzymic concentration in severe anemia is associated specifically with suppression or malfunction of the hematopoietic system. 8. It is believed that the relative concentration of enzyme can be used as a test of bone marrow function in severe anemias. It is hoped that further investigation may result in a useful test of wider application.

Description: 1. The subcutaneous administration of a single massive dose of cortisone to mature male albino mice produces: (1) A prompt and prolonged fall in the level of circulating lymphocytes and eosinophils which persists as long as nine days after the administration of the hormone. (2) A granulocytopenia which resolves at two days and is accompanied by a period of bone marrow blocking followed by resumption of delivery and myeloid hyperplasia. 2. No inhibitory effect of cortisone on cell proliferation is seen in the bone marrow. 3. There is no change in hemoglobin concentration following cortisone treatment. 4. Adult male albino mice are satisfactory animals for serial observations of the effect of cortisone on the circulating blood elements and the bone marrow.

Description: 1. Arteriovenous eosinophil differences across the spleen were measured in 10 dogs during the development of adrenal-mediated eosinopenia. In 7 instances the stimulus was epinephrine, in 2 ACTH, and in 1 operation alone. 2. In no case was significant net splenic uptake of eosinophils indicated. In 3 out of 7 epinephrine experiments and in 1 ACTH experiment, a significant net splenic output of eosinophils was found. For all 10 experiments averaged, the splenic output of eosinophils was significant. 3. Arteriovenous neutrophil and hematocrit differences were insignificant and were independent of simultaneous eosinophil differences. 4. It is concluded that the spleen is not a significant site of uptake on destruction of eosinophils during the development of adrenal-mediated eosinopenia. Under certain conditions, it actually extrudes eosinophils into the general circulation at a time when other factors are causing eosinophil disappearance. 5. The results suggest but do not establish the presence of a discriminatory splenic mechanism for the control of blood cells, capable of storing cells of one type while it releases others into the circulation.

Description: 1. The literature on eosinophil variations under various stresses is reviewed in the light of recent interest in the adrenal cortex as a regulator of blood eosinophil levels. Chamber eosinophil counts are necessary for accuracy in these studies. Phloxine-propylene glycol or eosin-acetone stains with standard white cell pipets and chambers may be employed for simple and satisfactory chamber counts. 2. Statistical formulae are applied to chamber eosinophil counts and marrow differential counts to delineate the theoretic errors. Actual observations differ slightly from the predicted results but substantiate the appreciable intrinsic errors frequently seen. 3. Studies on physiologic variation are reviewed. There are significant minute-to-minute fluctuations as well as a more marked diurnal trend in eosinophil levels with a mid-morning low and a night-time peak. Comparison of single morning eosinophil levels with single noon specimens may be misleading. 4. Oral ephedrine is a potent eosinopenic agent. 5. Repeated tests of the eosinopenic response to ephedrine or epinephrine frequently show significant variation in the same subject. 6. Many patients with various diseases fail to react adequately to ephedrine or epinephrine. 7. Repeated and controlled observations are necessary before conclusions regarding pituitary-adrenal function can be drawn from eosinophil counts alone.

Description: 1. Detailed hematologic observations, bone marrow aspirations and blood volume determinations were made on 20 patients with rheumatoid arthritis and allied disorders before, during and after the administration of either ACTH or cortisone. 2. Significant reticulocytosis occurred in every patient during therapy, but its magnitude was poorly correlated with either the initial degree of anemia or subsequent increase in circulating red cell mass. 3. There was an increase in hematocrit and total circulating red cell mass of all anemic patients who responded clinically to either ACTH or cortisone. There was little or no improvement of anemia when the clinical response was poor. 4. Polycythemia did not occur in any patient during prolonged therapy or with repeated courses of either ACTH or cortisone. 5. Hemodilution and hemoconcentration were much more profound during and after ACTH administration than they were with cortisone. 6. Bone marrow studies revealed moderate depression of the erythroid series before treatment. At the end of therapy erythroid elements were normal. 7. Significant polymorphonuclear leukocytosis occurred its all patients during therapy while lymphopenia was inconstant and unsustained. Circulating eosinophils were depressed more with ACTH than with cortisone treatment. 8. Before treatment eosinophils and their precursors were present in the bone marrow its normal or increased numbers. During therapy the number of these cells was unchanged in the marrow, even when there was profound peripheral eosinopenia 9. The role of ACTH and cortisone in the physiologic mechanism of hematopoiesis is discussed. 10. The improvement in the anemia associated with inflammatory disease in response to ACTH or cortisone therapy probably is a reflection of the control of the underlying disease rather than a primary "stimulation" of the bone marrow.

Description: 1. The hematologic findings on 21 patients who received fractionated intra-arterial HN2 therapy are presented. The arteries cannulized were the external carotid in 9 cases, the internal carotid in 4 cases, the external iliac in one case and the lower abdominal aorta in 7 cases. The total dose of drug given varied from 0.25 to 3.10 mg./Kg. 2. The effects on bone marrow and peripheral blood were essentially the same in character as those observed after intravenous administration of HN2. Larger doses of HN2 were tolerated when administration was intra-arterial. 3. A consistent early depression of marrow eosinophils was noted in all cases with original differential counts of 2 per cent or more, except in one patient who had Hodgkin’s disease and extensive psoriasis. 4. A differential effect of HN2 on marrow in the region of therapy and the marrow from other sites was demonstrated by simultaneous aspiration studies. 5. Factors reducing hematologic damage include occlusion of venous return from the injected site and prolongation of therapy. 6. Only one patient in this group developed severe hypoplasia of the bone marrow. Studies on this and other patients treated by the lower abdominal aortic route suggest that more than 1.1 mg./Kg. HN2 per course in this region may be hazardous. 7. Four patients died within two weeks following therapy with symptoms suggesting profound electrolyte disturbance. It is probable that limiting factors other than hematologic may occur with localized HN2 therapy.

Description: 1. Thin films of freshly drawn hemolymph of about 360 insects belonging to 61 different species were observed under the phase contrast microscope and the process of coagulation investigated. Control of the observations by motion picture recordings were performed in a few species. 2. In the hemolymph of all the insects investigated, a category of hyaline hemocytes can easily be recognized under the phase contrast microscope from the other kinds of blood cells. Attempts of identification of these hyaline hemocytes with elements described in the classifications of insect blood cells based on fixation and staining were inconclusive. 3. In the insects in which the blood clotting does not occur, these hyaline hemocytes do not exhibit any important alteration. In species inn which blood coagulation occurs, the cells of this category appear to be highly labile to contact to glass surfaces. They undergo rapid modifications in their structure which play an important if not exclusive part in the initiation of the plasma coagulation. In the present material these alterations are of two kinds, each of them being related to a different appearance of the plasma reaction. They can occur alone or together. 4. In contrast to the hyaline hemocytes, the other categories of blood cells do not take part in the process of coagulation. Scattered or agglutinated at random, they are passively embedded in the coagulum. 5. In the present material, hemolymph coagulation appears to be a continuous process, initiated by alterations taking place in a single category of hemocytes. These alterations are followed by various degrees of plasma coagulation, from a general macroscopic clotting to a limited reaction detectable only under the microscope. 6. The results are at variance with former data in which the coagulation of insect blood was described as being either a cellular agglutination, in which no special part was recognized to be played by a special category of cells, or a plasma coagulation, both considered as two physiologically distinct processes, which can occur independently or together. On the other hand, the present results show an analogy with the type of coagulation described in crustacean blood by Hardy, Tait, Tait and Gunn, in which a special category of cells, Hardy's explosive cells, as distinguished from the other blood cells, has a specific and important part to play.

Description: 1. A practical chamber method has been devised for enumerating the megakaryocyte concentration of aspirated bone marrow. 2. Marrow aspirations from 23 healthy young adults had a mean megakaryocyte concentration of 4.0 ± 1.1 per 10,000 total nucleated cells or 6.1 ± 1.6 per 10,000 granulocytes. 3. Of the three principal cell types in marrow, i.e. granulocytes, erythrocytes and lymphocytes, the megakaryocyte content seemed to parallel most closely the granulocytes.

Description: Human red cells treated with trypsin in such a way as to become agglutinable in the presence of incomplete antibodies are affected in certain other respects. Their volume is slightly increased, their ghosts are unusually rigid or "volume-occupying," their osmotic and mechanical fragilities are slightly increased and their electrophoretic mobility is reduced. These changes are probably due to effects on the protein components of the red cell surface ultrastructure. Similar effects are also produced by the related enzyme mexacain.

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: The alkaline and acid phosphatase content of blood and bone marrow has been studied by the methods of Gomori-Takamatsu and Gomori in normal individuals and in a great number of different diseases. The presence of a particular phosphatase is related to cellular reaction. Alkaline phosphatase was found in orthochromic normoblasts, polymorphonuclear leukocytes, metamyelocytes and older megakaryocytes. Acid phosphatase was demonstrated in basophilic normoblasts, polymorphonuclear leukocytes and their juvenile forms and in young megakaryocytes. In red blood cells and young megakaryocytes phosphatase was present chiefly in the nuclei, in leukocytes and older megakaryocytes in the cytoplasm. By studying erythropoiesis, especially during active regeneration of blood, it has been shown that alkaline phosphatase plays a decisive part in the disappearance of the normoblast nucleus. In certain diseases abnormality is observed in the disappearance of the normoblast nucleus (effort syndrome, rheumatoid arthritis). In others acid phosphatase occurs in large amounts in the youngest normoblasts (polycythemia vera, rheumatoid arthritis, nephritis). At the same time phosphatase may also be found in polymorphonuclear leukocytes and their juvenile forms. In rare cases of severe anemia alkaline phosphatase (panmyelophthisis) or acid phosphatase (achrestic anemia) is noted in large amounts in nucleated red cells and in leukocytes. Only alkaline phosphatase is present in polymorphonuclear leukocytes and metamyelocytes in leukemias, myeloma, Hodgkin’s and Paget’s disease, and in some malignant tumors of the skeleton and the liver. Several circumstances indicate that phosphatases might destroy cells or their nuclei. In some cases phosphatase activity is physiologic, in others phosphatase concentration is related to accelerated or malignant growth of bone marrow or tissue associated with blood formation. Finally there are cases in which there is as yet no explanation for the appearance of phosphatases.

Description: Periodic Rh antibodies were carried out over a period of one to four years in a series of 60 sensitized Rh-negative individuals. The results indicate that once an individual has become sensitized to the Rh factor, the antibodies persist in his body for the remainder of his life. An illustrative case is presented in which antibodies were found in the serum of an Rh-negative woman who had never received an injection of blood and who had had her last pregnancy twenty-two years previously.

Description: 1. Bitches with erythrocytes lacking the canine A factor were immunized with intravenous injections of A-positive dog cells and mated with A-positive sires. ll A-positive pups born to such dams developed hemolytic disease provided they suckled the immunized dam during the first day of life. There was no evidence of transplacental isoimmunization or of transfer of antibody across the placenta from mother to pup. Anti-A could not be demonstrated in the blood of pups at birth and was not acquired by pups that first suckled an immunized bitch after the first day of life. A-positive pups born to a non-immunized bitch developed hemolytic disease after suckling an immunized foster dam on the day of birth. 2. Of 24 affected A-positive pups, 3 were sacrificed, 9 died within three days of birth and 12 recovered, 2 with the aid of transfusions of A-negative blood. Autopsies revealed varying degrees of hepatomegaly, splenomegaly, erythroid hyperplasia of the bone marrow, extra-medullary erythropoiesis, and questionable evidence of specific injury of the nerve cells of the basal nuclei of the brain. 3. The degree of anemia in the A-positive pups varied widely, the minimum hematocrit for the group being 10 per cent at forty-eight hours after birth. Erythroblastosis, reticulocytosis and spherocytosis were noted in most of the severely affected pups and osmotic fragility of the red cells was substantially increased in all of the A-positive pups exposed to anti-A. 4. The concentration of bilirubin in the serum of most of the affected pups was only slightly increased. The relatively small increases in serum bilirubin, compared with those in hemolytic disease of human infants, are presumably attributable to the unusual capacity of the dog liver for excreting bilirubin. 5. The red cells of all affected A-positive pups gave antiglobulin (Coombs) reactions and in surviving pups the cells were agglutinable in antiglobulin rabbit serum for as long as twenty-two days in A pups and sixty-five days in A’ pups. The degree of reactivity with antiglobulin serum was not correlated with the severity of the hemolytic process. Erythrocytes of the very mildly affected A’ pups were strongly agglutinated by antiglobulin serum but showed no definite sphering and only slight increase in osmotic fragility. 6. Serum of the 20 A-negative litter mates examined in this study contained anti-A for periods as long as thirty-two days while the red cells of these pups remained normal. A few of the A-positive pups also had anti-A in the serum. The in vitro behavior of anti-A in the pups’ sera was identical with that of anti-A in the maternal sera. 7. Twelve C-positive pups in 2 litters born to bitches immunized against the canine C factor showed no evidence of hemolytic disease. 8. Hemolytic disease of newborn dogs is compared with that of human infants and the need for further investigation of certain aspects of the disorder in both species is stressed.

Description: Dilution with physiologic saline solution and other fluids accelerates the coagulation of properly collected normal and hemophilic blood and plasma in silicone coated vessels, with or without the aid of activating agents such as platelets, thromboplastin, cephalin, glass particles or plasma euglobulin fractions. When normal plasma is diluted under a concentration of about 20 per cent, its rate of clotting is prolonged, principally because of diminution in prothrombin. Regardless of the activating agent used, the rate of coagulation of hemophilic plasma can be made equal to that of normal plasma by appropriate dilution. These findings speak against the existence in hemophilic blood or plasma of a deficiency in any procoagulant factor, and support the concept of the presence in excess of a stabilizing inhibitor which slows the conversion of prothrombin to thrombin by one or both of the following mechanisms: (1) reducing or inactivating the effect of released coagulants (antithromboplastin activity) (2) conjugation with a procoagulant thereby maintaining it in an inactive form (anti Ac-globulin activity).

Description: The investigations reported in the present paper deal with the behavior of the peripheral blood and the bone marrow in urethane-produced agranulocytosis and panmyelopathy in the cat. The principal conclusions follow: 1. A severe panmyelopathy can be produced regularly in cats by the administration of relatively small doses of urethane: viz., 0.05 to 0.1 Gm. per Kg. per day. The panmyelopathy is still reversible if the urethane is stopped early enough and if the animal is maintained with antibacterial therapy. 2. The time required for recovery of the bone marrow after cessation of urethane administration depends on the total dosage of the drug. After small doses recovery occurs rapidly; after large doses much more slowly. Individual sensitivity of the experimental animal also plays a role, however. 3. The effect of smaller doses of urethane (0.05 Gm./Kg./day) on the bone marrow is to produce inhibition of maturation of granulocytopoiesis, and, to a lesser degree, of thrombocytopoiesis. To a still lesser extent, there is also inhibition of maturation of the red cell series. 4. The effect of larger doses of urethane (0.1 Gm./Kg./day) on the bone marrow, the effect on maturation is overshadowed by a generalized depletion of all marrow cells. Part of this effect is due to a decrease in the number of mitoses, with reduction of the mitotic index; as well as qualitative alterations in the mitotic patterns. It is probable that the effect on mitoses results in the formation of cells which are incapable of life as well as in an overall decrease in the formation of cells. 5. In the cat, the neutrophilic granulocytes are most sensitive to the action of urethane. The lymphocytes are next most sensitive; then the megakaryocytes, finally the erythroblasts. The eosinophils were remarkably insensitive. 6. The normal plasmacellular reticulum cells are relatively insensitive to urethane. This is in contrast to the neoplastic myeloma cells in human beings which have been reported to be sensitive to urethane. 7. The administration of folic acid or of vitamin B6 to the experimental animals had no effect on the course of urethane agranulocytosis. 8. The normal granulocytopoiesis of the cat is about as sensitive towards urethane as the pathologic granulocytopoiesis of chronic myeloid leukoses of human beings. This is in contrast to the findings in other animals, and to normal granulocytopoiesis of man. The variation in behavior possibly indicates variations in enzyme systems in the various types of granulocytopoiesis, with only certain enzymes being affected by urethane. 9. Further experiments are in progress to determine if the effect of urethane on the bone marrow can be counteracted by means of other growth substances. It is hoped that better insight may thus be gained into the mechanism of action of urethane.

Description: 1. Methods for continuous intervenous, arterial venous, and interarterial cross transfusions in man have been developed and a total of seven procedures have been successfully performed. 2. The interarterial method was preferred for an investigation of the pulmonary leukocyte removal mechanism and has been carried on for as long as twenty-six hours exchanging 150 liters of whole blood both to and from each participant. 3. On three occasions within twelve hours after the cross transfusions were terminated, a marked decrease in the leukocyte count occurred in 1 leukemic participant. Marked generalized improvement in the leukemic status occurred after each drop in the leukocyte count. 4. By cross transfusing an adult with myelogenous leukemia and a child with lymphogenous leukemia it was possible to pass myeloid cells through the pulmonary leukocyte removal mechanism into the circulation of the patient with lymphogenous leukemia. 5. An excessive leukocyte removal mechanism was demonstrated during another cross transfusion by a patient with sub-leukemic lymphogenous leukemia. 6. Cross transfusion in man is experimental and offers a technic of value as an investigative method for the study of formed elements and chemical constituents of the blood under these circumstances. 7. Careful cross matching for compatability of blood and Rh type is essential and the hazards and risks of the procedure have been emphasized.

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.

Description: 1. The effect of transfusion polycythemia upon bone marrow activity and erythrocyte survival has been studied in 4 normal young adult males. 2. Plasma volumes did not change significantly throughout the period of study. 3. Total red cell masses increased to the "expected" levels, i.e., to the total of the subjects’ cells plus the transfused cells immediately after the transfusions. Thereafter these fell progressively, reaching the control levels in about forty days. 4. Survival of the infused cells was normal. 5. The subjects’ own red cell masses fell progressively at first. This was due to decreased erythropoiesis rather than to increased destruction. This is indicated by (a) no consistent significant elevation in hemolytic indexes; (b) change of the myeloid-erythroid ratio from normal values of 4 to 7:1 to about 20:1 two weeks post-infusion; (c) consistent decrease in circulating reticulocytes. 6. The depression of erythropoiesis was directly related to the quantity of red cells infused; almost complete cessation of red cell synthesis followed an increase of the red cell mass by forty per cent. 7. The depression of erythropoiesis was only temporary. As soon as the total circulating red cell mass returned to the pre-injection level, erythropoiesis proceeded at a normal rate. 8. No consistent changes in the circulating white blood cells, totals and differentials were noted. 9. Mild abnormalities in some liver functions were observed. Whether these should be attributed to the effects of the transfusions directly, or to the mild febrile responses experienced by the subjects shortly after the infusions cannot be stated with certainty at present.

Description: 1. Two patients with a syndrome of hemolytic anemia, thrombocytopenic purpura and arteriolar occlusions are described. 2. In one patient, a long remission followed splenectomy, and improvement for ten days followed the administration of ACTH. No claim is made that either form of therapy induced the remission, but trial of these procedures in other cases is indicated. 3. Evidence is presented that the primary vascular lesion is a degenerative process in the arteriolar and capillary walls rather than thrombus formation in the lumen of the vessel. The occurrence of aneurysms in arterioles and precapillanes was a striking observation. 4. The histologic changes suggest a relationship to the so-called collagen group of diseases. 5. It is suggested that the term, thrombotic thrombocytopenic purpura, be retained until further study of the conditions permits application of a more suitable name.

Description: 1. A case of sickle cell anemia who had numerous hemolytic crises and also hypersplenic effects such as pancytopenia was treated by splenectomy with marked improvement in general condition and in the blood. The literature relating to this subject is reviewed. Those cases with the largest spleens appear to have shown the most striking degrees of improvement. 2. In cases of sickle cell anemia showing various indications of hypersplenism, and excessive hemolysis, splenectomy may be a rational therapeutic procedure.

Description: Simultaneous bone marrow smears and platelet counts on the peripheral blood were made preoperatively and on the third and the tenth days after operation in 14 patients. Observations of the activity of the megakaryocytes of the bone marrow on the third day were inconclusive and require further observation. There was, however, a significant increase in activity on the tenth day. It is postulated that the thrombocytosis which occurs on the tenth postoperative day is the result of increased activity of the megakaryocytes of the bone marrow induced by the attempt of the body to replenish depleted platelet reserves.

Description: 1. The original Arinkin technic of sternal puncture has been modified to include iliac, spinous process and tibial marrow aspiration. 2. There has been a need for improvement and standardization of the method of handling aspirated bone marrow. 3. An improved technic of marrow aspiration is described which combines direct Arinkin type smears with marrow particle and concentrate preparations. Fixed sections may also be obtained. An important innovation is the use of a dilute solution of sodium heparin in the aspirating syringe to prevent clotting so that all marrow obtained may be utilized. 4. The "pure" marrow fragment or particle smear is the most valuable for diagnostic purposes. Smears prepared from concentrated marrow cells and directly from the mixed aspirate are also studied routinely but are chiefly of value when marrow particle smears either can not be obtained or are unsatisfactory. Section of marrow particles is rarely indicated. Surgical trephine biopsy of the sternum is still performed when repeated aspirations fail to procure adequate marrow. 5. A gradual transition from the Arinkin method to the use of marrow fragments and concentrate smears is advisable because of the differences in cellular content and appearance of the preparations. A knowledge of the normal is essential. 6. The use of this combined technic of marrow aspiration has resulted in improved knowledge of normal and abnormal marrow obtained by the puncture method. It is of great value in the study of cells of the lymphocytic, plasmocytic and megakaryocytic series, and in routine search for the "L. E." cell. A higher degree of accuracy has been attained in diagnosis than was possible by the Arinkin technic. The procedure is especially valuable in the diagnosis of lymphosarcoma, plasma cell myeloma, thrombocytopenic purpura, leukopenic and pancytopenic states, hypoplastic and aplastic anemias and aleukemic leukemias.

Description: 1. Three patients with hereditary spherocytosis and 1 patient with the Fanconi syndrome (pancytopenia and multiple congenital abnormalities) were transfused prior to splenectomy with normal erythrocytes of types which could be differentiated serologically from those of the recipients. The proportions of donated and patient’s cells in peripheral blood and in blood washed from the minced spleens were determined by differential agglutination, and the osmotic and mechanical fragilities of the two types of cells in peripheral and splenic blood were measured by differential agglutination of the corpuscles remaining in each test tube after partial hemolysis had occurred. 2. In each case of hereditary spherocytosis the proportion of recipient’s cells was much higher in splenic than in peripheral blood, indicating selective retention of the thicker corpuscles within the spleen. Osmotic fragility of thie patient’s red cells was much greater in samples of splenic mince blood than in peripheral venous samples, while the fragility of the donated red corpuscles was normal or nearly normal in both splenic and peripheral blood. In the patient exhibiting the Fanconi syndrome, on the other hand, neither the patient’s red cells nor donated red cells were retained to any extent in the spleen and the fragility of neither type of cell was altered. 3. Spleens removed surgically from 3 patients with idiopathic thrombocytopenic purpura were perfused with mixtures of normal A or B cells and group O cells drawn from a splenectomized individual with hereditary spherocytosis. During perfusion the spheroidal cells were selectively removed from the mixtures and at the end of each experiment red cells of this type predominated in the blood samples washed from the minced spleens. A fourth excised spleen was perfused with a mixture of two types of normal cells, neither of which was retained to any extent by the spleen during perfusion. The perfusion experiments show that spleens from patients with nonhemolytic disease are also capable of selective trapping of spheroidal cells. 4. The experiments described indicate that the spleen acts as a filter and trap and as an "incubator" in accelerating destruction of red corpuscles in patients with hereditary spherocytosis.

Description: 1. A case of chronic hemolytic anemia, present since infancy, in a young Negro male has been reported. There were no hematologic abnormalities in the six family members who were examined. 2. The unusual features in this patient were normal red blood cell fragility, the absence of spherocytosis and marked punctate basophilia. These findings appear to conform to the syndrome of hereditary hemolytic anemia without spherocytosis described by Haden. This is the first reported case of this syndrome in a Negro. 3. Examination of material obtained by splenic puncture revealed that: a. Stippled cells were much fewer in the spleen aspirate than in the peripheral blood. b. Basophilic granules were found engulfed by the reticulo-endothelial cells of the spleen. These data suggest as a possible explanation for the hemolytic mechanism that the spleen selectively destroys the abnormal stippled erythrocytes. The mechanism of formation of these erythrocytes remains obscure.

Description: A simple but effective method of demonstrating "L. E." cells in the peripheral blood has been described.

Description: Clinical, hematologic and genetic data on 28 cases of hereditary spherocytosis are presented for the purpose of characterizing this disorder as completely as possible. On the basis of this experience it is recommended that the following typical laboratory findings be sought in establishing a diagnosis in suspected cases: (1) Presence of spherocytes or abnormally thick red cells in peripheral blood; (2) greater than normal osmotic fragility of the red cells; in cases in which the fragility of fresh cells is not significantly increased, determinations should be made after sterile incubation of the blood at body temperature for 24 hours; (3) greater than normal mechanical fragility of freshly drawn red cells; (4) negative antiglobulin (Coombs) test; (5) greater than normal lysis of the red cells during sterile incubation at body temperature for 48 hours; and (6) presence of similar abnormalities in relatives. Abnormality of the erythrocyte persisted in all of the 11 patients in this series followed one or more years after splenectomy. An unusual case of chronic hemolytic anemia is described but not included in the numbered series because (1) both parents were hematologically normal and (2) spherocytosis and abnormally great osmotic and mechanical fragility and autohemolysis could not be demonstrated after the fifth postoperative month. Classification of this case is deferred pending further experience. Demonstration in a parent, sibling or offspring of red cells showing the afore-mentioned abnormalities is necessary for an unequivocal diagnosis, but this requirement cannot always be met because relatives may not be available for examination. Moreover, when parents and/or several siblings are examined without positive findings, low gene expressivity, gene mutation and illegitimacy may be considered as explanations. Evidence is cited to suggest the possibility of a low degree of penetrance or expression in some cases and to illustrate the need for still more sensitive laboratory tests that might aid in diagnosis of the mildest forms of this disease. The lower incidence of spherocytosis in siblings of propositi than in offspring of propositi is cited as evidence bearing on the theory of gene mutation in some propositi. A simplified "qualitative" test of osmotic fragility of incubated red cells is described.

Description: Saccharated iron oxide has been given to 23 patients in doses of 480 to 1,430 mg. Toxic symptoms were minimal. Hematologic response was good in 21 patients. Continued bleeding accounted for the poor response in 2 persons. Saccharated iron oxide is a safe preparation for clinical use. It is easy to administer. Further clinical evaluation is warranted. Excessive amounts of intravenous iron should not be given to any individual.

Description: Using the Nadi reaction, cytochemical studies of the cytochrome oxidase activity of cells of lymphoid and myeloid tissue were carried out. Normal cells and those from patients with Hodgkin’s disease and leukemia were examined. With the exception of monocytes and macrophages cells from lymph nodes, spleen, bone marrow and peripheral blood show a low level of cytochrome activity when compared with myocardial, liver and renal tubular epithelium. Leukemic cells and those from lymph nodes affected by Hodgkin’s disease contain about the same degree of cytochrome activity as their normal counterparts, under the conditions of this study. The cytoplasmic particles stained by the Nadi reaction correspond in size, distribution and number to particles which can be stained supravitally by janus green.

Description: 1. Cerebral blood flow and metabolism were measured in 16 patients with pernicious anemia. Seven of the patients were restudied in various stages of therapy. 2. The patients fell into two equal groups, those with severe anemia and those with moderate or no anemia. In the first group, cerebral blood flow was increased and cerebral vascular resistance decreased; in the second group, cerebal blood flow was decreased and vascular resistance increased. In both groups, cerebral oxygen and glucose consumption was decreased, as was cerebral venous oxygen tension . 3. There was a good correlation between the mental status defects and cerebral oxygen consumption and between severity of neurologic involvement and cerebral oxygen consumption. There was no correlation between cerebro-vascular resistance and cerebral oxygen consumption, nor between degree of anemia and cerebral oxygen consumption. 4. Specific therapy resulted in a moderate increase in cerebral oxygen consumption and cerebro-vascular resistance. In no instance did cerebral oxygen consumption become normal. 5. The disparity between the functional ability of the patients and the low values for cerebral metabolism is discussed. 6. It is concluded that pernicious anemia results in specific nervous system involvement not related to the anemia, and that this damage is at least partially irreversible in many patients.

Description: The authors report a fatal case of essential pulmonary hemosiderosis in a 5 year old boy. The child showed typical signs of the disease with relapsing pulmonary attacks with hemoptysis and pronounced anemia of iron deficiency type. Huge amounts of hydrolyzable iron were found in the lungs.

Description: When used in equimolar concentrations, five different calcium-combining anticoagulants produced uniform dilution of a given blood sample. The potassium anticoagulants seem to exert a hemolytic effect on the erythrocytes. The plasma dilution caused by a given concentration of the calcium-combining anticoagulants ranged from 8 to 20 per cent in the bloods examined. This shows the importance of using serum or unoxalated plasma for comparing the plasma protein concentrations of individual blood samples.

Description: 1. A method has been presented for the quantitative enumeration of the bone marrow cells by adjusting the differential count for the degree of cellularity, which is derived from imprint preparations of femoral marrow. 2. This method has been employed in a study of bone marrow changes at various intervals following 700 r total body x-irradiation in the rat. The findings have been correlated with the peripheral blood counts. 3. On the basis of numerical changes and morphologic observations, the mechanism of the effects of irradiation on the various cell series of bone marrow is evaluated. The erythroid cells show rapid decreases in number due to their marked destruction. The myeloid cells chiefly undergo accelerated maturation resulting in increased segmented forms in both blood and bone marrow. The decreases in megakaryocytes, mast cells and lymphocytes appear to be related to their life spans in the absence of further production. The reticulum cells and plasma cells show neither an absolute increase nor decrease. 4. Formation of new cells, except possibly for plasma cells, from the reticulum cells is completely inhibited for the first nine days following total body irradiation of rats with 700 r. Regeneration is first apparent at twelve days after x-ray as evidenced by areas of erythropoiesis, and to a lesser extent, myelopoiesis. Megakaryocytes reappear in small numbers at fifteen days. Although this regeneration is apparent in only 50 per cent of the animals studied at twelve and fifteen days following irradiation, all these animals showed evidence of transition of reticulum cells to mast cells.

Description: 1. In the usual therapeutic dosage in mice with the Ak4 strain of leukemia, TEM caused a prolongation of survival time, held the total leukocyte count down at severely leukopenic levels, delayed the appearance of immature lymphocytes in the peripheral blood, and caused a marked delay in the appearance of leukemic infiltrations of the liver, spleen, bone marrow and kidney. 2. A single massive dose of TEM, administered on the seventh day of the disease when the total leukocyte count was high, caused a rapid fall in the count and depressed the number of prolymphocytes in the peripheral blood. 3. Single massive doses of such structurally related compounds as hexamethylene diethylenurea, 2,4-diethylenimino-6-amino-s-triazine, and to a lesser degree, 2-ethylenimino-4,6-dimethoxy-s-triazine caused a similar fall in the total leukocyte count when administered on the seventh day of the disease.

Description: 1. Vitamin B12a, derived from vitamin B12 by catalytic hydrogenation, is as potent a hematopoietic agent as vitamin B12 when administered parenterally to patients with pernicious anemia in relapse. 2. The hematopoietic activity of vitamin B12a, like that of vitamin B12, and of vitamin B12b, is potentiated by simultaneous oral administration with normal human gastric juice. 3. Observations on one patient suggest that vitamin B12a, like vitamin B12, will arrest the progress of subacute degeneration of the spinal cord in pernicious anemia.

Description: 1. A simplified microspectrograph for use in the visible range is described. 2. A nonrecording microdensitometer for use with the above is discussed. 3. The Soret band of hemoglobin is demonstrable in individual erythrocytes and is not affected by fixing or staining. 4. The ratio of the hemoglobin concentration to cosin staining is constant in erythrocytes and the cytoplasm of immature red cells but not in the nucleus. 5. There is no detectable hemoglobin or verdoperoxidase in the granules present in the eosinophile granulocytes of the camel. 6. Both dried and fixed and stained erythrocytes can take up carbon monoxide from the atmosphere.

Description: 1. Twelve cases of immunization by pregnancy or blood transfusion in which the Rhesus antibody anti-E was found, to the exclusion of anti-D or any other Rh antibody are reported. 2. Two hundred and fifty Rh antibodies were discovered in this laboratory during 1949 in 37,972 antenatal cases not previously tested, i.e., 1 in 152. Seven of these were pure anti-E, i.e., 2.8 per cent. This frequency is considerably higher than any previously reported. 3. All of the anti-E sera tested possessed to some degree, and 5 of them to a marked degree, the property of distinguishing cells with a double from those with a single dose of the antigen E. 4. The presence, in cells of various genotypes, of tine antigen D in association with the antigen E reduced their agglutinability with the same anti-E serum in such a manner as to suggest that the agglutinability of these cells depends upon the relative quantities of the E and D anitigens present. 5. By means of parallel titrations with a genotyping anti-E serum, against cells of known and unknown genotypes, it was possible to distinguish E/E from E/e cells in the presence or absence of associated D and, as a corollary, to distinguish D/D from D/d cells in the presence of associated E/E or E/e. 6. These results fit the hypothesis that the specific Rhesus antigens C, D and E are produced from a limited amount of "basic raw material" by the activity of the three genes: that the genes act independently and in competition and that the amount of each antigen produced is proportional to the dosage and "competitive value" of the corresponding gene.

Description: Two cases previously diagnosed as aplastic anemia were found to have abnormal susceptibility of the red blood cells to hemolysis after chilling and after addition of acid in the presence of their own as well as control serum. The serum of the patients had no such effect on control erythrocytes. This erythrocyte abnormality was accompanied by a severe anemia of the macrocytic type, slight reticulocytosis, depression of granulocytes and platelets, and erythropoietic hyperactivity of the bone marrow in the early stages and hypoactivity in the late stages. Increased blood destruction was evidenced not by jaundice, but by persistent hyperhemoglobinemia in the serum and increased iron excretion in the urine, without nocturnal intensification. Chilling in vivo produced hemoglobinuria in Case 1, but only slight evidence of increased blood destruction in Case 2. Administration of large doses of ammonium chloride failed to produce hemoglobinuria in either case, although evidence of an increase in blood destruction was detectable. Both patients had historical and serologic evidence of syphilis. Autopsy of Case 1 showed, in addition to syphilitic aortitis, marked hemosiderosis of the kidneys, liver, spleen and lymph nodes. These were clearly not cases of paroxysmal hemoglobinuria é frigore (Donath-Landsteiner). They resembled cases of chronic hemolytic anemia with nocturnal paroxysmal hemoglobinuria (Marchiafava-Micheli), but the erythrocyte fragility to chilling, the absence of nocturnal increase in the hemolytic process and the presence of syphilis rendered the acceptance of such diagnosis difficult.

Description: Observations of thick smears of marrow obtained from the femur, tibia, humerus, sternum, and ribs (in rats), showed that in the first three bones, the number of mast cells was much higher than in the latter two bones. A method for the quantitative estimation of the total number of nucleated cells and of mast cells in the bone marrow of rats is presented. The method involves dilution of the marrow, in a red cell pipet, with a 1:50,000 solution of toluidine blue in 3 per cent acetic acid. This method confirmed the results of mast-cell distribution obtained in the study of marrow smears.