bluuebunny/crossref_metadata_2025 huggingface dataset:
Storage Cells of Spleen and Bone Marrow in Thalassemia: An Ultrastructural Study
Authors: Beltrami C. A. , Bearzi I. , Fabris G.
Published: October 2019

In the spleens and bone marrows of patients with thalassemia, we have consistently observed PAS-positive histiocytes that contain numerous cytoplasmic bodies surrounded by a single membrane and filled with bundles of fine fibrils (typical thalassemia storage cells). In addition to the characteristic cytoplasmic bodies, some histiocytes contained leukocytes, erythrocytes, platelets, and plasma cells in different stages of digestion (transitional cells). These findings suggest that the storage cells in thalassemia are macrophages that are unable to metabolize completely the products of phagocytized blood cells. In the spleen the histiocytes were closely associated with plasma cells. When stained by the Giemsa method, the storage cells appeared as ‘sea-blue histiocytes." The histologic, histochemical, and ultrastructural features distinguish the storage cells of thalassemia from Gaucher’s cells and from storage cells of other disorders in which there is excessive destruction of formed elements of the blood, i.e., chronic myelogenous leukemia and idiopathic thrombocytopenic purpura.
Leukocyte Interferon Production, RNA Synthesis, and PHA Response in Patients With Infectious Mononucleosis
Authors: Pidot Anne L. Rassiga , Maurer L. Herbert , McIntyre O. Ross
Published: October 2019

Sequential measurements of in vitro Newcastle disease virus-induced interferon (IF) synthesis, spontaneous RNA synthesis, and PHA-induced blastic transformation were made in leukocytes derived from ten patients with infectious mononucleosis (IM). The median leukocyte IF production on the day of diagnosis was found to be significantly depressed, 35 international units (IU) IF per 2 x 106 leukocytes, in comparison to a median titer of 463 IU after recovery (p < 0.002) or to a median of 406 IU found in 52 cultures from normal healthy individuals. No spontaneous production of leukocyte IF or circulating serum IF was noted. An increased rate of spontaneous leukocyte RNA synthesis during the first 2 hr of culture was noted in nine of the ten subjects (p < 0.001). Four of the ten patients showed a markedly defective response to PHA after 96-144 hr in culture. A suggestive, but not statistically significant, correlation was noted between increased spontaneous RNA synthesis, defective late PHA transformation, and defective virus-induced IF production. All defects persisted during the 1-wk study period and were resolved after recovery. We conclude that major temporary alterations occur in several parameters of leukocyte function in IM.
Cytogenetic Studies in Myeloma
Authors: Dartnall Jean A. , Mundy G. R. , Baikie A. G.
Published: October 2019

Chromosomes have been studied in cultures of nucleated peripheral blood cells in 30 patients with myeloma. Both unstimulated cultures and cultures stimulated by phytohemagglutinin (PHA) were examined in all cases. Mitoses were observed in 13 unstimulated cultures of peripheral blood from 12 cases, providing evidence for the presence of circulating abnormal cells in these patients. In 19 stimulated cultures of peripheral blood from 17 cases, abnormal mitoses were present, seven of these included abnormalities involving a chromosome of group 13-15. In five cases, a large acrocentric chromosome was observed similar to that previously described in some cases of myeloma. In eight cases, chromosome studies were also carried out on bone marrow to allow comparison of methods. Examination of stimulated peripheral blood cells was found to be the better method. Marked aneuploidy was not a feature of the results. No correlation was found between the cytogenetic abnormalities and previous treatment, disease stage, further progression of the disease, or serum and urinary protein abnormalities. The results are discussed, particularly with reference to technical factors influencing the frequency of demonstrable chromosome abnormalities in these cases and the relative merits of examination of peripheral blood and bone marrow.
Measurement of Serum Folate Levels and Serum Folic Acid-binding Protein by 3H-PGA Radioassay
Authors: Waxman Samuel , Schreiber Carol
Published: October 2019

A radioassay for the measurement of serum folate levels using commercially available beta lactoglobulin, as the folate-binding protein, and 3H-pteroylglutamic acid (3H-PGA) is reported. The assay was run in a one-step simultaneous addition at room temperature. A standard curve was constructed to a sensitivity of 0.25-10 ng of N-methyltetrahydrofolic acid (methyl-THFA). There was a clear correlation with separation into normal (greater than 6 ng/ml), indeterminate range (3-6 ng/ml), and deficient (0-3 ng/ml), as measured by radioassay. Serums from patients receiving antibiotics had normal folate levels with this assay. 3H-PGA was also used to measure serum folic acid-binding protein (FABP). In normal serums, the mean FABP was 18 pg bound/0.4 ml serum, while in folate-deficient serums it was 133 pg bound/0.4 ml of serum. Folate-deficient patients had a fall in serum FABP to the normal range when treated with folic acid. Serum FABP, in a patient on a folate-deficient diet, increased with early folate deficiency and abruptly fell to normal with a regular diet. FABP in lysates of folate-deficient bone marrow was higher than normal marrow. Patients with B12 deficiency, multiple myeloma, cirrhosis, pregnancy, or taking oral contraceptives had normal FABP. Elevated FABP was found in two out of ten patients taking Dilantin. This radioassay and the measurement of FABP should simplify the diagnosis of folate deficiency.
Differences in Inducing Activity for Human Bone Marrow Colonies in Normal Serum and Serum From Patients With Leukemia
Authors: Mintz Uri , Sachs Leo
Published: October 2019

Normal serum and serums from patients with acute and chronic leukemia were assayed for granulocyte colony-inducing activity with human bone marrow cells. Serum from untreated acute leukemia, but not from the other patients, showed about normal inducing activity at low serum concentration and lower than normal activity at high concentration. This suggests that serum from patients with acute leukemia contained an inhibitor for colony formation. Serums from chronic myeloid leukemia were in about the same range as normal, whereas serums from chronic lymphocytic leukemia showed the highest colony-inducing activity.
5-Azacytidine: A New Active Agent for the Treatment of Acute Leukemia
Authors: Karon Myron , Sieger Lance , Leimbrock Suzanne , Finklestein Jerry Z. , Nesbit Mark E. , Swaney Jerry J.
Published: October 2019

Thirty-seven children with acute leukemia were treated with 5-azacytidine in 5-day courses given every 14 days. Six out of 14 children with acute myelogenous leukemia who were adequately treated achieved an M1 marrow. Five of these subsequently developed complete remissions lasting 8 mo, 6 mo, 3 mo, 2 mo, and 2 mo. Of 22 children with acute lymphocytic leukemia, one achieved an M1 marrow and one an M2 marrow. The former attained a complete remission which lasted 3 mo. The maximum tolerated dose is between 150 to 200 mg/sq m on a daily x 5 schedule given every 14 days. The impressive activity of 5-aza-C in patients with acute myelogenous leukemia resistant to cytosine arabinoside indicates that this drug will become an important addition to the therapeutic armamentaria against this type of leukemia.
"Foreign Serum" Heterophile Antibodies in Patients Receiving Antithymocyte Antisera
Authors: Pirofsky Bernard , Ramirez-Mateos J. C. , August Anne
Published: October 2019

Forty-six patients with venous immunologically mediated or lymphoproliferative diseases received goat antihuman thymocyte antisera globulin (ATG) therapy. Heterophile antibody production occurred in 91 % of these patients, following ATG administration. The heterophile antibody was absorbable with both guinea pig kidney and boiled beef erythrocyte antigens. This corresponds to a form of heterophile antibody previously reported as a "serum sickness" variety. The heterophile antibody was composed of a mixture of gamma G and gamma M antibodies and was partially absorbed by ATG. Serum sickness did not occur in any of these patients, in spite of the production of heterophile antibodies. It is recommended that the involved heterogenetic antigen be termed "foreign serum" variety, and the resulting antibody described as a "foreign serum" type of heterophile antibody.
Cholinesterase as a Possible Marker for Early Cells of the Megakaryocytic Series
Authors: Jackson Carl W.
Published: October 2019

Among marrow cells of rats only megkaryocytes stain appreciably for acetylcholinesterase. An investigation was therefore undertaken to determine whether this staining characteristic might be used to identify precursors of recognizable megakaryocytes. Marrow cells smaller than recognizable megakaryocytes were positive for the enzyme. The number of these cells increased after induction of severe thrombocytopenia by injection of antiplatelet antiserum. Subsequently, the number of recognizable megakaryocytes increased, suggesting their derivation from the small acetylcholinesterase-positive cells. These results suggest that acetylcholinesterase activity can be used as a marker for megakaryocytic cells that cannot be detected with conventional staining methods.
A Hypertransfused Mouse Assay for Thrombopoietic Factors
Authors: Cooper George W. , Cooper Barbara , Ossias A. Lawrence , Zanjani Esmail D.
Published: October 2019

A method was developed for the quantitative separation of platelets from CF1 mouse whole blood. This made it possible to determine the platelet incorporation of 35S-sulfate without the necessity of doing platelet counts. Daily hypertransfusions of the mice to three to four times normal platelet levels for 4-5 days significantly reduced platelet uptake of radiosulfate to an average of about 40% of the nontransfused controls. Mice rendered thrombocytopenic 48 hr earlier by antiplatelet serum, had 2-day 35S uptakes over 2 1/2 times the controls and 6 times the hypertransfused animals. The administration of a total of 2 ml of serum, given twice daily for 3 days from a thrombocytopenic patient with Hodgkin's disease caused a highly significant 103% rise in radiosulfate incorporation when compared with saline in the hypertransfused mouse. Normal human serum from a healthy donor caused a small and insignificant rise. The serum from a patient with Hodgkin's disease caused a highly significant 63% rise in 35S incorporation when compared to the normal serum.
Autoantibodies in Acquired Hemolytic Anemia With Special Reference to the LW System
Authors: Vos G. H. , Petz L. D. , Garratty G. , Fudenberg H. H.
Published: October 2019

Most autoantibodies in patients with warm antibody autoimmune hemolytic anemia (AIHA) have specificity within the Rh system. Using rare cells such as -D- and Rhnull cells, Weiner and Vos (1963) described specificity against normal cells (nl), partially deleted cells (pdl), and deleted cells (dl). Recently, autoantibodies which failed to react with Rhnull cells that were of anti-U specificity have been described. It was suggested that the "Rh related" autoantibodies that cannot be identified as specific Rh antibodies may be anti-U. In the present study we examined eluates from the red cells of eight patients with AIHA using a panel of extremely rare cells and cross-absorption and elution techniques. We demonstrated autoantibody specificities not definable without the rare cells and, further, defined heterogeneity of the LW antigen. Autoantibodies with U specificity occurred in three eluates only. It was always present with an antibody of another specificity. Six of the eluates contained anti-LW, two anti-nl, five anti-pdl, three anti-dl, and one anti-e. Absorption and elution studies using the rare Rh-positive LW-negative (Mrs. Bigelow) showed that anti-pdl may in fact represent anti-LW + LW1 and that Mrs. Bigelow may represent a weak variant of LW. Injection of her red cells into guinea pigs produced an anti-LW that reacted similarly to the antibody produced by injecting Rh-positive LW-positive cells. An analogy to the ABO is suggested that normal Rh-positive LW-positive cells represent LW1, Rh-negative LW-positive cells represent LW2, Mrs. Bigelow represents LW3 and Rhnull cells represent the only true LW-negative (lw).
The Effects of Injection of Human Factor VIII Antibody Into Rabbits
Authors: Shen S. M-C. , Feinstein D. I. , Rapaport S. I.
Published: October 2019

Rabbits were injected with an immunoglobulin fraction of human serum containing a factor VIII antibody. Factor VIII levels fell abruptly, persisted below 10% of a rabbit plasma standard for 12 hr, and returned to normal by 120-168 hr. The factor VIII antigen-antibody reaction did not result in Intravascular clotting as evaluated by kinetic studies with 125I-fibrinogen. However, small falls in factor V and factor VII levels were observed over a 6-hr period after the injection. Platelets fell to about one-half of initial values within 15 min, rose to 80% of initial levels over 2 hr, and subsequently declined to 65%-70% of initial levels. WBC levels fell to below 20% of initial values 2 hr after the injection but returned to about 75% of initial values by 6 hr. Total hemolytic complement activity was unaffected. Animals made granulocytopenic with nitrogen mustard and animals with hereditary C'6 deficiency behaved similarly to normal animals. One may conclude that the injection of human factor VIII antibody into rabbits produces a rabbit model with impaired intrinsic coagulation suitable for studies of the mechanism of endotoxin-induced intravascular clotting.
Intravascular Clotting After Endotoxin in Rabbits With Impaired Intrinsic Clotting Produced by a Factor VIII Antibody
Authors: Shen S. M-C. , Rapaport S. I. , Feinstein D. I.
Published: October 2019

A rabbit model in which intrinsic clotting was selectively impaired by injection of a human factor VIII antibody was used to evaluate the mechanism of endotoxin-induced intravascular clotting in cortisone-treated rabbits. Three groups of animals were studied: a control group given factor VIII antibody followed by saline; a second control group given an inert material followed by endotoxin; and an experimental group given factor VIII antibody followed by endotoxin. The following parameters were measured: 125I-fibrinogen kinetics, fibrinogen levels, factor VIII, factor VII, factor V, WBC, platelets, and hematocrit. The kidneys were examined for deposition of fibrin. Mean values for factor VIII at the time of injection of the second test material and mean values for fibrinogen consumed in the 6 hr after the second injection were as follows: antibody-saline group, 8.5% and 11.0 mg/kg; control material-endotoxin group, 90% and 29.6 mg/kg; and antibody endotoxin group, 7.0% and 32.7 mg/kg. Factor V, factor VII, granulocytes, and platelets fell in both groups of animals given endotoxin. One animal in each group given endotoxin developed gross renal cortical necrosis. These data establish that selective impairment of the intrinsic clotting reactions does not reduce the amount of clotting induced by a single injection of endotoxin in the cortisone-treated rabbit.
Kinetics of Lymphocytes in Chronic Lymphocytic Leukemia: Studies Using Continuous 3H-Thymidine Infusion in Two Patients
Authors: Theml H. , Trepel F. , Schick P. , Kaboth W. , Begemann H.
Published: October 2019

Two patients with typical chronic lymphocytic leukemia received a continuous infusion of 3H-thymidine for 7 days. The production rate of lymphocytes was low in the lymph nodes, the marrow, and the blood. The fraction of newly formed lymphocytes appearing in the blood per day was only about 0.5%. The absolute production rate of blood lymphocytes however was very high, 690 cells/cu mm blood/day in one patient and 725 cells/cu mm/day in the other. The daily accumulation of newly produced lymphocytes in the circulating blood was estimated to be 3.0 x 109 cells. Most of these newly formed cells were lost from the blood lymphocyte compartment within 3 wk. They are considered to be short-lived lymphocytes with half-lives of 3-8 days. About 90% of the blood lymphocytes were long-lived with a turnover time in excess of 1 yr in the blood lymphocyte compartment. The short-lived cells were both large and small lymphocytes, the long-lived cells were almost exclusively small lymphocytes. Lymphoid cell proliferation was virtually absent in the blood, very low in the marrow, and marked in the enlarged lymph nodes. The exchange of lymphocytes seemed to be partially intact between lymph nodes and blood but more or less completely blocked between marrow and blood.
Tissue Factor Activity of Normal and Leukemic Cells
Authors: Garg Sudershan K. , Niemetz Julian
Published: October 2019

Generation of procoagulant activity was studied in leukocytes and lymphocytes from normal human peripheral blood, leukocytes from patients with chronic myeloid leukemia (CML), chronic lymphatic leukemia (CLL), rabbit peripheral leukocytes, and macrophages harvested from the peritoneal cavity of rabbits. Marked procoagulant activity developed in peritoneal macrophages and rabbit peripheral leukocytes when incubated with endotoxin. Human peripheral blood leukocytes and lymphocytes also generated procoagulant activity. When endotoxin was omitted the procoagulant activity generated was considerably (3-12 times) lower. Leukocytes from patients with CML were less active than normal leukocytes. Leukocytes from patients with CLL generated very little procoagulant activity even after prolonged incubation. An antibody to rabbit tissue factor neutralized the tissue factor activity generated by rabbit leukocytes.
Immunologic Studies of Antihemophilic Factor (AHF, Factor VIII). V. Immunologic Properties of AHF Subunits Produced by Salt Dissociation
Authors: Rick Margaret E. , Hoyer Leon W.
Published: October 2019

Human antihemophilic factor (AHF, factor VIII), a large plasma protein with a molecular weight of approximately two million, is dissociated by changes in ionic strength. The immunologic properties of subunits obtained by sucrose density ultracentrifugation in 1 M NaCl and by agarose gel filtration in 0.24 M CaCl2 have been determined using human and rabbit anti-AHF. Asymmetric dissociation of AHF has been identified with formation of two subunits in these separations: a nonfunctional highmolecular-weight (HMW) subunit similar in size to plasma AHF which is identified by immunoprecipitation and radioimmunoassay for AHF antigen, and an active lowmolecular-weight (LMW) subunit which is not detected by these antigen assays. The LMW subunit retains AHF antigens, however, for it is inactivated by both human and rabbit anti-AHF. Antibody neutralization studies verify the presence of AHF antigens on the HMW subunit. These immunologic studies provide constraints which must be incorporated into models of AHF structure.
Inhibition of Human Platelet Aggregation by Plasmin Digests of Human Factor VIII
Authors: Donati M. B. , de Gaetano G. , Vermylen J. , Verstraete M.
Published: October 2019

Human factor VIII, upon prolonged incubation at 37°C with plasmin, inhibits platelet aggregation induced by ADP, adrenaline, or collagen. The human factor VIII digests are not anticoagulant in the thrombin time or partial thromboplastin time. Factor VIII split products may interfere in vivo with platelet function and thus could contribute in some way to the hemorrhagic diathesis of fibrinolytic states.
Lymphocytes Forming Red Cell Rosettes in the Cold in Patients With Chronic Cold Agglutinin Disease
Authors: Feizi Ten , Wernet Peter , Kunkel Henry G. , Douglas Steven D.
Published: October 2019

Lymphocytes that formed rosettes with human red cells in the cold were observed in the peripheral blood of four patients with chronic cold agglutinin disease. Similar rosettes were not observed in normals or other controls studied. Specificity for I and i red cells was demonstrated which paralleled that of the serum macroglobulins. The rosettes were inhibited by soluble I and i antigens in accordance with the specificity of the serum antibodies. Lymphocytotoxicity assays with idiotypic antisera made to the serum macroglobulins detected the same proteins on the lymphocyte surface. Evidence was obtained that the rosette test underestimated the number of cells bearing the specific cold agglutinin protein. Antisera against cold agglutinins as well as anti-IgM antibodies blocked the rosette formation. By all of these criteria, the surface lymphocyte receptors were indistinguishable from the serum antibodies of these patients.
A "Philadelphia-like" Chromosome Derived From the Y in a Patient With Refractory Dysplastic Anemia
Authors: Warburton Dorothy , Bluming Avrum
Published: October 2019

A small acrocentric chromosome, identical in appearance to the Philadelphia chromosome of chronic myelocytic leukemia, was identified in all karyotyped marrow cell metaphases prepared from a 74-yr-old male with refractory dysplastic anemia. The abnormal chromosome was shown by quinacrine fluorescent staining to be derived from a Y chromosome which has lost the characteristic brilliantly fluorescent material of its long arms. A normal Y chromosome was consistently observed in karyotypes of cultured peripheral blood lymphocytes from this patient.
Change in Marrow and Spleen CFU Compartments Following Leukemia Virus Infection: Comparison of Friend and Rauscher Virus
Authors: OKunewick James P. , Phillips Eleanore L.
Published: October 2019

The effects of the Friend and Rauscher leukemia viruses on the marrow and spleen colony-forming unit compartments have been studied. It was found that the two viruses produced qualitatively similar but quantitatively different effects relative to each other. Comparison of the effects of the individual virus on spleen with those on the marrow also showed striking differences. In the spleen by 14 days after infection with either virus the CFU compartment was grossly enlarged. In the marrow of the Friend-infected mouse the CFU number was not significantly different from normal, and in the Rauscher-infected mouse there was a 50% drop in marrow CFU. Measurement of the F factor showed that infection by either virus resulted in colony-forming units that were deficient as compared to the normal in their ability to successfully "seed" in the spleens of irradiated normal recipients, with the greater depression in F factor being found in cells obtained from spleen tissue. These results indicate that in both cases the nature of the host tissue (marrow versus spleen) exerts a significant modifying effect on the expression of the virus action, and also suggest that the pluripotent colony-forming unit may be a target cell for these viruses.
Biological Effects of Repeated Leukapheresis of Patients With Chronic Myelogenous Leukemia
Authors: Vallejos C. S. , McCredie K. B. , Brittin G. M. , Freireich E. J.
Published: October 2019

Fifteen patients with chronic myelogenous leukemia were managed only with repeated leukapheresis for up to 26 mo. At each procedure approximately 10 liters of blood were processed with a continuous-flow blood cell separator over a 3-hr period. Five patients had intermittent leukapheresis (1-2 procedures/wk), and ten of them had one or more series of intensive leukapheresis (4-5 procedures/wk). Addition of hydroxyethyl starch to the extracorporeal circuit was found to increase the removal of leukocytes fourfold. With intensive leukapheresis the leukocyte count decreased 80%, and the platelet count decreased 54% (mean values). With intermittent leukapheresis the corresponding figures were 70% and 35%. Thrombocytopenia was never clinically significant. All 15 patients experienced symptomatic improvement, and those with organomegaly had decrease in the size of the spleen and liver. Leukapheresis was not associated with morbidity, except that anemia became more severe and required periodic transfusions of packed red blood cells. Patients managed with leukapheresis did not enter bone marrow remission, and transformation of CML into blastic crisis was not prevented or delayed.
Iron in the Duodenal Mucosa of Normal, Iron-loaded and Iron-deficient Rats
Authors: Mattii Roberto , Mielke C. Harold , Levine Peter H. , Crosby William H.
Published: October 2019

Iron absorption studies with oral 59FeSO4 were performed on 13 iron-depleted, 11 iron-loaded, and 10 control rats, and nonheme iron was determined in both isolated epithelial cells and defoliated mucosa obtained from the duodenum. Mean absorption by the control animals was 18.5% of the dose. Both iron-depleted and iron-loaded groups showed significant differences in iron absorption (54.5% and 2.2% respectively). Compared with the normal controls, iron was decreased in the epithelial cells of the iron-deficient group, whereas higher concentrations were observed in the defoliated mucosa of iron-loaded animals. The latter observation was confirmed by the presence of iron-laden macrophages seen in sections of the lamina propria of the iron-loaded rats.
Detection of Myeloma Cells in the Urine Sediment
Authors: Pringle Jane P. , Graham Richard C. , Bernier George M.
Published: October 2019

The urine sediments of 18 patients with multiple myeloma were stained with tetrachrome stain and examined by light microscopy. Eleven of the eighteen sediments contained cells with the morphologic characteristics of plasma cells. When stained with fluoroscein conjugated rabbit immune globulin prepared against κ and λ light chains, eight of these sediments contained fluorescent staining plasma cells. In each case the cells stained with the antibody to the light chain of the patient’s myeloma protein and not with the other light-chain antibody. Examinations of sediment from patients with other types of renal disease and normal subjects did not reveal the presence of fluorescent staining cells. In the urine of two myeloma patients, cells were identified as plasma cells by electron microscopy. No correlation was found between the presence of plasma cells and the patient’s monoclonal protein, light-chain type, or the presence or degree of renal disease. The finding of myeloma cells in the urine presumably indicates an extramedullary extension of the disease, and their presence may contribute to the renal impairment commonly found in myeloma patients.
The Effect of Neutropenia on the Cell Cycle of Granulocyte Precursors in an In Vivo Culture System
Authors: Niskanen Eero , Tyler W. S. , Symann Michael , Stohlman Frederick , Howard Donald
Published: October 2019

Proliferation of myeloid elements, macrophages, and pluripotent stem cells (CFU) in diffusion chamber cultures of normal mouse bone marrow was examined. The relative growth of differentiated cells and CFU was clearly inhibited when the concentration of cells cultured was increased from 105 to 20 x 105 per chamber. When chambers were implanted into neutropenic hosts the growth of CFU and granulocytic cells was enhanced. The generative cycle of the earliest recognizable granulocytic cells, as estimated by flash labeling with tritiated thymidine, was comparable to that seen in normal bone marrow and was significantly shortened by culture in a neutropenic environment. Growth of pluripotent stem cells and myeloid elements in these cultures appears to be influenced both by short-range factors related to cell concentration and by long-range humoral factors.
Homozygous State for Hb Constant Spring (Slow-moving Hb X Components)
Authors: Eng Lie-Injo Luan , Ganesan J. , Clegg J. B. , Weatherall D. J.
Published: October 2019

A 12-yr-old Malay boy who was studied because his youngest brother and both his parents had the slow-moving Hb X components (earlier reported to lead to Hb H disease when combined with α-thalassemia) was found to be homozygous for the same slow-moving components. He had splenomegaly and a just palpable liver, mild anemia with microcytosis, hypochromia, slight morphologic changes of the red blood cells, and slight reticulocytosis. Of eight children in the family, six had the trait for the abnormality, one was normal, and one, the propositus, was homozygous. Structural studies of the isolated abnormal hemoglobin showed it to be identical to Hb Constant Spring (Hb CoSp), an α-chain variant with 172 residues instead of the usual 141, the additional 31 being attached to the C-terminal end. In addition to the abnormal α variant for which the propositus was homozygous, he also had normal Hb A and normal Hb A2 with normal α-chains. If the theory that Hb CoSp is due to a structural mutation affecting the terminator codon is correct, this case provides evidence for a duplication of the gene for α-chain production. Results of study of several erythrocyte enzymes are also reported.
Decreased Glutathione Peroxidase Activity Secondary to Severe Iron Deficiency: A Possible Mechanism Responsible for the Shortened Life Span of the Iron-deficient Red Cell
Authors: Rodvien Robert , Gillum Amanda , Weintraub Lewis R.
Published: October 2019

Red cell glutathione peroxidase activity decreased to 26% of initial activity following the induction of severe iron deficiency in rabbits. The fall in enzyme activity does not appear to be related to anemia per se, aging of the animals during the course of the experiment, or a generalized decrease in enzyme protein synthesis. With iron repletion, glutathione peroxidase activity did not return to initial values for 10 wk after the hemoglobin concentration had returned to normal. During this period of time, the young red cell population had a higher concentration of GSH-Px activity than the older cells. In contrast, there is no age differential in the concentration of this enzyme in normal red cells, and during the induction of iron deficiency young red cells have decreased enzyme activity as compared to older cells. Reduction in glutathione peroxidase activity may result in increased oxidative damage to the iron-deficient red cell. This, in turn, may be responsible for the previously reported membrane abnormalities and shortened red cell survival of iron-deficient red cells. It remains to be shown if glutathione peroxidase is dependent on iron for its synthesis or if the function of the enzyme is dependent on an iron-containing protein serving as an electron carrier.
Erythrocyte Coproporphyrin and Protoporphyrin in Ethanol-induced Sideroblastic Erythropoiesis
Authors: Ali M. A. M. , Sweeney G.
Published: October 2019

Sideroblastic erythropoiesis is a recognized complication of acute alcoholism1 and disturbances in porphyrin synthesis have been reported in sideroblastic anemia2. In order to investigate the mechanism of alcoholic-induced sideroblastic erythropoiesis coproporphyrin (C.P.) and protoporphyrin (P.P.) levels were measured in 26 patients admitted with acute alcoholism. Bone marrow examination showed that 11 patients had ring sideroblasts, while in 15 other patients there were adequate iron stores, but no ring sideroblasts. The red blood cell (C.P.) and (P.P.) levels were elevated in all of the 11 patients with ring sideroblasts, but in only three out of the 15 patients with no ring sideroblasts. These data clearly separate those patients with a disturbance of mitochondrial iron metabolism and increased erythrocyte porphyrins from other patients suffering from a comparable degree of alcoholism in whom neither abnormality appeared to exist. We conclude therefore that alcoholic sideroblastic erythropoiesis probably reflects ethanol-induced disturbance in the heme synthetic pathway.
Androgenic Hormones and Human Granulopoiesis In Vitro
Authors: Rosenblum Alan L. , Carbone Paul P.
Published: October 2019

Androgenic steroids in physiologic concentrations were found to stimulate human granulopoiesis in vitro. Testosterone in high concentration, but probably within the range achievable after intensive treatment, was found to inhibit granulopoiesis. Analysis of growth kinetics suggested that increased granulopoiesis results from proliferation of committed granulocyte precursors in the marrow rather than stem-cell activation.
The Effect of Methotrexate on Transformation and Mitosis of Normal Human Blood Lymphocytes In Vitro
Authors: Rozenszajn L. A. , Radnay J.
Published: October 2019

The effect of methotrexate (MTX) on transformation and mitosis of normal lymphocyte cultures was investigated. Mitosis was 1000-fold more sensitive to MTX than was transformation; mitosis could be prevented without any observable effect on transformation. The antimitotic effect of MTX depended on its concentration, duration of contact with lymphocytes, as well as on the developmental stage of the lymphocytes at the time of exposure. Incorporation of labeled precursors into protein, DNA, and RNA proceeded at normal rates unaffected by the presence of MTX. Transformation was partially inhibited by high concentrations of MTX (above 75 µg/ml). Folinic acid abolished the antimitotic effect of MTX, with the number of cells undergoing division directly proportional to the quantity of folinic acid added.
Hemoglobin Affinity for Oxygen in Chronic Renal Disease: The Effect of Hemodialysis
Authors: Lichtman Marshall A. , Murphy Marion S. , Byer Barbara J. , Freeman Richard B.
Published: October 2019

The affinity of hemoglobin for oxygen may increase significantly in subjects who are hypophosphatemic and alkalotic. We studied the organic phosphate content and oxygen binding by hemoglobin of red cells in subjects undergoing hemodialysis, during which time a decrease in plasma inorganic phosphate and an increase in blood pH may occur. Red cell 2,3-DPG was not correlated with plasma inorganic phosphorus, whereas red cell ATP was highly correlated with plasma inorganic phosphorus when analyses were made on predialysis samples. Predialysis red cell inorganic phosphorus was highly correlated with plasma inorganic phosphorus, supporting the concept that intraerythrocytic inorganic phosphorus is maintained by a gradient from plasma to cell. Plasma inorganic phosphorus decreased by 45% during the period of hemodialysis, whereas red cell inorganic phosphorus did not change. Red cell 2,3-DPG, ATP, and oxygen binding by hemoglobin at standard conditions of temperature, pH, and pCO2 were not altered after 6 hr of hemodialysis. Plasma pH and base excess increased during dialysis. The increase in base excess, an estimate of the non-pH-dependent effect of CO2 on oxygen binding by hemoglobin, counterbalanced a portion of the effect of elevated pH on hemoglobin— oxygen affinity under in vivo conditions. Hence, only a slight increase in oxygen binding by hemoglobin occurred. Moreover, late dialysis symptoms were not associated with the degree of alkalosis or with the extent of change in hemoglobin’s affinity for oxygen. Red cell 2,3-DPG content was lower and hemoglobin’s affinity for oxygen was higher in subjects with chronic renal disease than in nonazotemic subjects with similar hemoglobin deficits. Moreover, increased red cell ATP in chronic renal disease patients did not influence oxygen binding by hemoglobin.
Hepatic Erythropoietin Production in the Lead-poisoned Rat
Authors: Schooley J. C. , Mahlmann L. J.
Published: October 2019

Extrarenal erythropoietin production is increased above normal in lead-injected rats exposed to hypoxia either immediately after or 1 day after nephrectomy. Extrarenal erythropoietin production in lead-poisoned rats is actinomycin D sensitive, and the major extrarenal site of production or activation of the hormone in normal or lead-poisoned anephric rats is the liver.
MU-Chain Disease in an African Patient
Authors: Bonhomme J. , Seligmann M. , Mihaesco C. , Clauvel J. P. , Danon F. , Brouet J. C. , Bouvry P. , Martine J. , Clerc M.
Published: October 2019

A new case of µ-chain disease occurring in an African patient is described. The main clinical feature was a massive splenomegaly. Contrary to five previously reported cases, this patient was not affected with chronic lymphocytic leukemia, and vacuoles were not found in the cytoplasm of the bone marrow plasma cells. A considerable increase of the plasma volume was noted without hyperviscosity. The electrophoretic pattern showed an abnormal band in the α2 region. The anomalous protein was devoid of light chains and was present in the form of disulfide-linked polymers of incomplete µ-chains. The molecular weight of the monomer was approximately 58,000. The protein comprised the Fc fragment and a part of the Fd segment. Bence Jones protein was not found in the urine, and light chains were not present in the cells producing the µ-chain disease protein.
The Use of 125I as a Membrane Protein Label for Erythrocyte Survival Studies
Authors: Weintraub Martin , Gerson Kenneth , Silber Robert
Published: October 2019

Radioactive iodide, incubated with erythrocytes in the presence of lactoperoxidase and hydrogen peroxide, is covalently bound to membrane proteins. Using this technique, rabbit erythrocytes were labeled with 125I, reinjected into the donors, and erythrocyte survivals determined. The values for the erythrocyte half-lives and the shape of the decay curves were comparable to those reported using DF32P. Following hemolysis of erythrocytes double labeled with 125I and 51Cr, radioactivity from both these isotopes appeared mainly in liver, lung, kidney, spleen, and urine. Enzymatic iodination provides a noneluting label of erythrocyte membrane proteins for the study of survival, sequestration and turnover of this red cell component.
Some Properties of the Circulating Hemopoietic Stem Cells
Authors: Gidáli Julia , Fehér I. , Antal Sára
Published: October 2019

Some properties of hemopoietic stem cells (CFU) circulating in the peripheral blood of the normal mouse (PCFU) were studied. Those parameters were chosen that are well-known characteristics of bone marrow stem cell populations. Leukocytes separated from the peripheral blood were the source of PCFU. The radiosensitivity (D0 value), growth curve, seeding efficiency (f number), and the turnover state (number of PCFU in S phase measured by 3H-thymidine killing technique in vitro) of PCFU were compared to the same parameters for bone marrow CFU. Several characteristic changes in these parameters were found for PCFU: (1) Lower radiosensitivity (D0 = 140 R), (2) Higher seeding efficiency (f number 20.9%), (3) Higher sensitivity to 3H-thymidine in vitro (killing effect 32%). The doubling time was the same both for CFU and PCFU (21 hr), but the logarithmic growth of the PCFU population started by approximately 36 hr later than that of the marrow-derived CFU population. All these findings support the idea that PCFU represent a subpopulation of the heterogeneous bone marrow CFU population.
Platelet Coagulant Activities and Hemostasis: A Hypothesis
Authors: Walsh Peter N.
Published: October 2019

Platelets have recently been shown to participate in reactions with blood coagulation factors at every stage of intrinsic clotting, from contact activation to fibrin formation. Platelets can trigger intrinsic coagulation by two alternative pathways, the first of which involves factors XII and XI and adenosine diphosphate, and the second of which bypasses factor XII, provided factor XI and collagen are present. Additional evidence indicates that subsequent coagulation reactions occur on the platelet surface, where active clotting factors are protected from inactivation by naturally occurring inhibitors. Based on these observations, an hypothesis is presented in which the events of primary hemostasis (platelet adhesion, aggregation, and release) and blood coagulation are linked. As platelets aggregate to form a hemostatic plug, they provide a protective and catalytic surface for activation of the clotting mechanism and fibrin formation. Localized hemostasis is promoted and circulating blood kept fluid by means of a number of control mechanisms, some of which are mediated by autocatalytic effects of thrombin.
A Comparison of the Behavior of 111In and 59Fe-labeled Transferrin on Incubation With Human and Rat Reticulocytes
Authors: Beamish Michael R. , Brown Elmer B.
Published: October 2019

The uptake by human and rat reticulocytes of 111In and 59Fe bound to transferrin has been studied. The results indicate a significant difference between the behavior of the two isotopes in both human and rat incubation mixtures. Reticulocyte uptake of 111In from human and rat serum was 30% and 12% that of the 59Fe after a 30-min incubation. The process was temperature dependent, inhibited by sodium arsenite, and related to the reticulocyte percentage of the cells in the reaction mixture. Washed reticulocytes, previously incubated for 30 min with either 59Fe or 111In bound to serum were reincubated in unlabeled serum. Up to 85% of the 111In label and less than 10% of the 59Fe on the reticulocytes were released on reincubation, indicating that, in contrast to 59Fe, the majority of the 111In label remained membrane bound. Specific binding of unlabeled In transferrin was demonstrated by its inhibitory effect on 59Fe-transferrin uptake by rat reticulocytes. Both In- and Fe-transferrin were found to have similar binding affinities for the receptor sites. The 111In remaining in lysates obtained from washed reticulocytes after incubation with 111In-labeled serum and reincubation in unlabeled serum did not appear to be associated with either the hemoglobin or heme molecules.
The Role of Peroxidase in the Bactericidal Activity of Human Blood Eosinophils
Authors: Bujak Joseph S. , Root Richard K.
Published: October 2019

Utilizing cell preparations that contained greater than 90% eosinophils and sodium azide to inhibit the lysosomal enzyme peroxidase, studies of eosinophil bactericidal activity were undertaken. Eosinophils did not kill bacteria as well as neutrophils, principally because of a reduced phagocytic capacity. In contrast to neutrophils, eosinophils had high resting iodination activity but failed to increase this activity following phagocytosis. One millimolar sodium azide, which inhibits both eosinophil and neutrophil peroxidase as measured by the iodination of trichloroacetic acid precipitable protein, impaired neutrophil staphylocidal activity, but enhanced eosinophil killing of Staphylococcus aureus primarily by increasing phagocytic uptake. Bactericidal studies utilizing lysostaphin suggest that neutrophil peroxidase exerts its contribution to bactericidal activity only during the early postphagocytic period. Eosinophil peroxidase is genetically and biochemically distinct from neutrophil peroxidase, and appears to play no role in the bactericidal activity of intact eosinophils.
Granulocyte Transfusions in Leukopenic Dogs: In Vivo and In Vitro Function of Granulocytes Obtained by Continuous-Flow Filtration Leukopheresis
Authors: Debelak Kim M. , Epstein Robert B. , Andersen Burton R.
Published: October 2019

The present studies were carried out to (1) evaluate a leukoadhesive technique for obtaining granulocytes for transfusion, (2) assess the granulocytes by in vitro techniques, and (3) determine the efficacy of granulocyte transfusion in preventing sepsis in leukopenic dogs. Dogs were rendered transiently leukopenic (< 500 per cu mm) by intravenous cyclophosphamide, 40 mg/kg. Quantitative and qualitative blood cultures were obtained from all animals until death or hematologic recovery. Granulocytes were obtained on nylon filters by a continuous flow system and eluted with an ACD plasma saline solution. Granulocyte function was studied in vitro by chemotaxis, phagocytosis, intracellular killing, and electron microscopy. In vivo studies consisted of the measurement of granulocyte increments in transfused leukopenic dogs, T ½ of infused granulocytes, and protection of transfused dogs from septicemic episodes. Eluted granulocytes, when compared to normal controls, showed reduction in in vitro functions. These functions improved in granulocytes isolated post-transfusion from recipient dogs. An average of 3 x 1010 granulocytes could be obtained during a 1-hr leukopheresis of normal donors. Increments in recipient dogs averaged 2590 per cu mm. Five nontransfused leukopenic dogs developed septicemia and died within 7 days. Six dogs were treated with infusions of granulocytes. Three recovered completely, and three died of thrombocytopenic hemorrhage with negative blood cultures. One dog showed a transiently positive blood culture that became negative following transfusion. Septic episodes were significantly reduced in granulocyte transfused dogs (p < 0.01). It was concluded that continuous-flow leukofiltration yielded granulocytes in sufficient number and with adequate functional capabilities to provide significant protection against septic death in the leukopenic host.
Dysplastic Platelets and Circulating Megakaryocytes in Chronic Myeloproliferative Diseases. I. The Platelets: Ultrastructure and Peroxidase Reaction
Authors: Maldonado Jorge E. , Pintado Tomás , Pierre Robert V.
Published: October 2019

A light and electron microscopic study was made of the circulating platelets from five patients with variants of the chronic myeloproliferative diseases (agnogenic myeloid metaplasia, chronic granulocytic leukemia, and "megakaryocytic leukemia"). Morphologic abnormalities were found in at least some of the platelets in all of the patients. Presence of giant forms, paucity or absence of granulation, disorganization and scarcity of the microtubules, and haphazard distribution and hypertrophy of the dense tubular and open canalicular systems constituted the main abnormalities. Additionally, there were features of platelet immaturity, including abundant rough endoplasmic reticulum and ribosomes and presence of Golgi profiles, centrioles, and nuclear remnants. The relationship, if any, of these morphologic abnormalities to the hemostatic defects in myeloproliferative diseases remains to be elucidated.
Dysplastic Platelets and Circulating Megakaryocytes in Chronic Myeloproliferative Diseases. II. Ultrastructure of Circulating Megakaryocytes
Authors: Maldonado Jorge E.
Published: October 2019

Small circulating megakaryocytes were identified in four patients with chronic myeloproliferative disorders, and their morphologic features by light and electron microscopy are described. The morphologic and cytogenetic studies suggest that at least some of these cells are truly mononuclear. The circulating megakaryocytes were associated with the presence of dysplastic platelets. Both findings are thought to represent involvement of the megakaryocytic-platelet line, probably as an expression of stem cell disease.
Clonal Origin of the Philadelphia Chromosome From Either the Paternal or the Maternal Chromosome Number 22
Authors: Gahrton G. , Lindsten J. , Zech L.
Published: October 2019

The heteromorphic regions of chromosome 22 were studied with the quinacrine mustard fluorescence technique in eight patients with Philadelphia chromosome-positive chronic myelocytic leukemia and in their parents. The fluorescence pattern showed that the Philadelphia chromosome had originated from the paternal chromosome 22 in one case and the maternal in another. The other six cases were noninformative in this respect. The fluorescence pattern was consistent between cells in both the informative cases. These results speak in favor of a clonal origin of the Philadelphia chromosome from either the paternal or the maternal chromosome 22.