bluuebunny/crossref_metadata_2025 huggingface dataset:
Observations on the Effects of Vitamin B12, Liver Extracts, Folic Acid and Thymine on the Maturation of Megaloblasts in Culture
Authors: THOMPSON R. B.
Published: October 2019

Folic acid has been demonstrated to have a direct maturing effect on cultures of megaloblasts obtained from patients suffering from pernicious anemia in relapse. No effect has been observed with either liver extracts, vitamin B12 or thymine.
Acanthrocytosis
Authors: SINGER KARL , FISHER BEN , PERLSTEIN MEYER A.
Published: October 2019

1. Clinical and laboratory studies are reported on a 13½ year old boy, the offspring of a consanguinous marriage, who, after having been afflicted with a celiac syndrome in early childhood, developed a progressive ataxic neuropathy associated with a peculiar malformation of most of his circulating erythrocytes. Similar observations were previously described in the literature in two siblings also of consanguinous parents. The latter patients, however, had in addition, retinitis pigmentosa.

2. The erythrocytic anomaly consisted of an unusual type of "crenation"; the deformed red cells showed several, irregularly spaced, large and coarse projections on their surface which varied in width and length. Many of the cells resembled spherocytes with pseudopods. The term acanthrocyte (thorny red cell) is proposed for this particular type of misshapen erythrocyte.

3. The acanthrocytes exhibited a slightly decreased osmotic, a markedly increased lysolecithin and mechanical fragility, hut a normal heat and acid fragility.

4. The hemolytic index was within normal range, a finding which seems to preclude the existence of an exaggerated hemolytic process in vivo.

5. It is suggested that acanthrocytosis is genetically conditioned, and due to a mutant recessive allele for a gene which controls the normal architecture of the red cell.

6. Further observations are required to establish whether the association of acanthrocytosis, celiac disease in early childhood, and ataxic neuropathy with or without retinitis pigmentosa constitutes a new hereditary syndrome.
Resistance of Reticulocytes and Young Erythrocytes to the Action of Specific Hemolytic Serum
Authors: CRUZ W. O. , JUNQUEIRA P. C.
Published: October 2019

A photoelectric method is used to determine the degree of hemolysis produced by specific hemolytic serum on erythrocytes of dogs, taking as a standard the amount required for 50 per cent hemolysis. The requirement of the normal dog red blood cell is 1.2 cu. ml. (±0.7 cu. ml.) of hemolytic serum, under the conditions described in the method.

The red cells of dogs given a sufficient dose of acetylphenylhydrazine to produce a severe anemia, followed by an intense period of blood regeneration, required an amount of hemolytic serum about two to four times greater than in the normal animal (values from 3.0 to 4.8 cu. ml. of hemolytic serum are recorded). This result shows that the reticulocytes and young erythrocytes are much more resistant than the normal adult red cell to the action of hemolytic serum. It was possible by this technic to concentrate the reticulocytes in vitro, destroying selectively the adult erythrocytes of a sample of blood by an appropriate amount of specific hemolytic serum.

The relationship between these results and the high reticulocytosis observed during crisis in hemolytic jaundice is discussed.
A Method for Separating and Concentrating Platelets from Normal Human Blood
Authors: MINOR ALLEN H. , BURNETT LEE
Published: October 2019

A method is described for the preparation of platelet concentrates from pint volumes of normal human blood collected in standard ACD bottles. Erythrocytes are removed by sedimentation with high molecular weight dextran, and platelets are concentrated by centrifugation in the presence of a surface-active agent, which permits their resuspension in a small volume of physiological fluid following prolonged centrifugation. The procedure results in the recovery of about 80 per cent of the total platelet complement of the blood. The platelets obtained are discrete and morphologically intact, and induce normal prothrombin consumption and clot retraction when added to platelet-poor plasma or thrombocytopenic blood. Transfusions of platelet concentrates given to thrombocytopenic patients have been followed by hemostatic effects lasting from 1 to 5 days.
Plasma Transfusions in Hemophilia
Authors: VAN CREVELD S. , PAULSSEN M. M. P.
Published: October 2019

Transfusions of heparinized plasma have a greater and more lasting effect on the coagulation time of hemophiliacs than transfusions of citrated plasma. Both in vitro and in vivo, heparinized plasma causes in hemophiliacs a far greater consumption of prothrombin as determined with the two-stage method than citrated plasma. In using the one-stage method no important differences in prothrombin activity are found after transfusions of heparinized and of citrated plasma respectively. This fact was thought to be connected with the more or less rapid appearance of an accelerator. Its a hemophiliac with a circulating anticoagulant, transfusions of heparinized plasma were unable to shorten the coagulation time to any important degree, nor did these transfusions cause an important decrease of serum prothrombin as determined by the two-stage method.
The Effect of a Folic Acid Antagonist, A-Methopterin, on the Level of the Circulating Eosinophils in Humans
Authors: WRIGHT JANE C. , HJELT INGA
Published: October 2019

1. Daily eosinophil counts were obtained on a group of 10 normal individuals. This control group showed a moderate daily fluctuation in the count, with only 1 exception.

2. Daily eosinophil counts were obtained on 16 chronically ill or convalescent patients to whom A-Methopterin was administered. The eosinophil curves of 10 of these patients showed a fall of over 50 per cent at or prior to A-Methopterin toxicity. In 1 case, the count increased then decreased at toxicity to the initial level. In 3 cases definite evidence of toxicity was not demonstrable and the eosinophil level did not decrease.

3. Six patients showed a fall in their eosinophil counts before clinical evidence of A-Methopterin toxicity. Three patients showed a fall in their eosinophil counts of 75 per cent at the time of A-Methopterin toxicity.

4. Twelve of 16 cases (75 per cent), to whom A-Methopterin was administered showed definite falls in the curves of daily eosinophil counts. That cortisone and the adrenocorticotropic hormone do likewise suggests, but is not evidence, that the mechanisms are related.
The Effects of Cobalt, Liver Extract and Vitamin B12 on the Anemia Induced by Hypophysectomy in Adult Female Rats
Authors: CRAFTS ROGER C. , TURNBULL DOROTHY S. , WEBER PATRICIA R.
Published: October 2019

Hypophysectomized adult female rats of the Wistar strain were treated with daily subcutaneous injections of 0.1, 0.3 and 0.5 mg. of cobalt nitrate for 50 days. Most striking results were obtained with the largest dose. The erythrocyte count in the hypophysectomized rats was elevated from a preoperative level of 7.9 million cells per cu. mm. to 11.6 million cells; the hematocrit, from 45.8 per cent to 60.8 per cent; and the hemoglobin, from 16.0 Gm. per 100 cc. to 21.0 Gm. Normal animals responded in a similar manner. Cobalt nitrate, therefore, not only prevented anemia but actually induced a marked elevation in the number of erythrocytes and in hemoglobin. Vitamin B12 and liver extract were ineffective.
Familial Hypoplastic Anemia with Congenital Abnormalities (Fanconi’s Syndrome)
Authors: REINHOLD J. D. L. , NEUMARK E. , LIGHTWOOD R. , CARTER C. O.
Published: October 2019

The published examples of Fanconi's syndrome are reviewed. The syndrome consists of familial hypoplastic anemia and is associated with pigmentation, microcephaly, strabismus, mental backwardness and other congenital anomalies. Two further examples occurring in a family of 7 children are described. Both sisters have macrocytic hypoplastic anemia, microcephaly and convergent strabismus; one has been treated by splenectomy. Three other siblings have microcephaly and other congenital abnormalities, but no anemia. There was no evidence of hemolysis. This condition is probably determined by a recessive gene.
Case Report: A Case of Erythremic Myelosis (Di Guglielmo’s Anemia)
Authors: MACKENZIE IAN , STEPHENSON A. G.
Published: October 2019

A case of erythremic myelosis is described which was characterized by a severe anemia from the beginning. There was irregular remitting fever, splenomegaly and hepatomegaly. The peripheral blood showed immature erythroblasts, granulopenia and thrombocytopenia. The bone marrow showed hyperplasia of erythropoiesis with maturation arrest, and a persistent megaloblastic type of nucleus in abnormal red cells. There was also a well defined proliferation of the reticulo-endothelial system. The course was that of the subacute variety of erythremic myelosis, but it began with an acute phase and ended acutely.

A remission of nearly three months without transfusion followed a transfusion of fresh blood. While this may be coincidental with a spontaneous remission we feel that it is probable that a maturation factor in the fresh blood was responsible. For this reason we feel it is reasonable to suppose that other cases of erythremic myelosis will benefit more from fresh blood than from stored blood; and, as in our case, the inevitable fatal outcome after two months or less can be postponed for an appreciable number of months.
Heparin-induced thrombocytopenia: laboratory studies
Authors: Kelton JG , Sheridan D , Santos A , Smith J , Steeves K , Smith C , Brown C , Murphy WG
Published: October 2019

This report describes studies into the pathophysiology of heparin- induced thrombocytopenia. The IgG fraction from each of nine patients with heparin-induced thrombocytopenia caused heparin-dependent platelet release of radiolabeled serotonin. Both the Fc and the Fab portions of the IgG molecule were required for the platelet reactivity. The platelet release reaction could be inhibited by the Fc portion of normal human or goat IgG, and patient F(ab')2, but not F(ab')2 from healthy controls. These results suggested that the Fab portion of IgG binds to heparin forming an immune complex and the immune complexes initiate the platelet release reaction by binding to the platelet Fc receptors. To directly challenge this hypothesis, we preincubated the serotonin-labeled platelets with the monoclonal antibody against the platelet Fc receptor (IV.3). This monoclonal antibody completely inhibited the release reaction caused by heparin and patient sera, as well as heat aggregated IgG, but did not block collagen or thrombin- induced platelet release. Heparin-dependent platelet release also could be inhibited in vitro by the addition of monocytes and neutrophils, but not by red cells, presumably because the Fc receptors on the phagocytic cells have a higher binding affinity for IgG complexes than do platelets. Platelets from patients with congenital deficiencies of specific glycoproteins Ib and IX (Bernard-Soulier syndrome) and IIb and IIIa (Glanzmann's thrombasthenia) displayed normal heparin-dependent release indicating that the release reaction did not require the participation of these glycoproteins. These studies indicate that heparin-induced thrombocytopenia is an IgG-heparin immune complex disorder involving both the Fab and Fc portion of the IgG molecule.
A monoclonal antibody to exfoliated surface vesicles that recognizes a membrane-associated erythroid burst-promoting activity
Authors: Dainiak N , Warren G , Sutter D , Kreczko S , Howard D
Published: October 2019

A monoclonal antibody (MoAb) recognizing a membrane-associated erythroid burst-promoting factor was prepared by immunizing BALB/c mice with plasma membrane-derived vesicles exfoliated from lymphocytes under serum-free conditions. Hybrids secreting antibody reactive with lymphocyte plasma membranes were formally cloned and IgG was purified from monoclonal supernatants or from BALB/c mouse ascites fluid. Two clones (D3-E4 and D3-G9) were found to suppress burst forming unit- erythroid (BFU-E) proliferation when added directly to serum-free human marrow culture. Inhibition to a level of 100% was observed in a dose- dependent fashion over a wide range of antibody concentrations (0–200 micrograms/mL). Neither antibody altered the proliferation of colony forming unit-granulocyte macrophage (CFU-GM) or colony forming unit- granulocyte-erythroid-monocyte-megakaryocyte (CFU-GEMM) progenitor cells in human bone marrow culture. The D3-E4 clone was found to produce an IgG1 antibody which adsorbs an erythroid burst-promoting activity (BPA) from supernatants of, and octylglucoside extracts of shed vesicles present in, serum-free, lymphocyte conditioned medium (LCM), and which recognizes a vesicular protein of Mr approximate 30,000 on immunoblots of membrane proteins electrophoresed on sodium dodecyl sulfate (SDS)/polyacrylamide and transferred to nitrocellulose. In contrast, the D3-G9 clone was found to produce an IgG1 cytotoxic antibody. These antibodies will be important to the study of cell-cell and growth factor-cell interactions in vitro.
Comparison of influenza A virus and formyl-methionyl-leucyl- phenylalanine activation of the human neutrophil
Authors: Hartshorn KL , Daigneault DE , White MR , Tuvin M , Tauber JL , Tauber AI
Published: October 2019

Influenza A virus (IAV) activates the human neutrophil, but induces a dysfunctional state as well. Cell activation may contribute to the containment of the virus and/or cause local tissue damage. Certain features of the neutrophil activation response elicited by IAV are distinctive when compared with that triggered by formyl-methyl-leucyl- phenylalanine (FMLP). An atypical respiratory burst response occurs in which hydrogen peroxide, but no superoxide, is formed. This unusual respiratory burst stoichiometry persists despite marked priming of the IAV-induced response. A comprehensive examination of the activation cascade initiated by these stimuli failed to show an explanation for these differences. Both IAV and FMLP comparably stimulate inositol trisphosphate and phosphatidic acid production. The subsequent increase in intracellular calcium (Ca2+i) upon FMLP stimulation was more dependent on extracellular Ca2+ than with IAV activation, but both stimuli induced Ca2+ influx. FMLP and IAV exhibited equal susceptibility to inhibition by protein kinase inhibitors in eliciting the respiratory burst, and actin polymerization occurred in response to each agonist. A possible explanation for the anomalous respiratory burst induced by IAV is that O2- is generated at an intracellular site inaccessible to assay, and/or virus binding to sialic acid constituents of the plasma membrane alters the O2- generating capacity of the respiratory burst oxidase; evidence for each mechanism is offered.
Growth in children after bone marrow transplantation: busulfan plus cyclophosphamide versus cyclophosphamide plus total body irradiation
Authors: Wingard JR , Plotnick LP , Freemer CS , Zahurak M , Piantadosi S , Miller DF , Vriesendorp HM , Yeager AM , Santos GW
Published: October 2019

Growth was assessed during the first and second years following bone marrow transplantation (BMT) in 47 children treated by either busulfan plus cyclophosphamide (BU/CY) (n = 24) or cyclophosphamide plus fractionated total body irradiation (CY/TBI) (n = 23). Before transplant, the median height was only 0.2 SD below age- and sex- adjusted means (range, -2.5 to +3.0). Height was greater than 2.0 SD below normal in only three patients (6%). The pretransplant heights were comparable in the BU/CY and CY/TBI groups (-0.1 v -0.6 SD, P = .35). Following transplant, median 1- and 2-year heights were 0.7 and 0.9 SD below normal, respectively. Growth rates were 2.2 SD and 1.4 SD below normal during the first and second years, respectively. Growth rates were greater than 2.0 SD below normal in 24 of 47 (51%) at 1 year and in 12 of 31 (39%) at 2 years after transplant. Growth rates in patients treated with BU/CY were comparable to those treated with CY/TBI during both years: -2.5 versus -1.7 SD during the first year (P = .19, Wilcoxon), and -1.5 versus -1.1 SD during the second year (P = .61). Growth rates during the second year correlated with growth rates during the first year (r = .36, P = .046). Growth rates during the first year were lower in patients who had been given prior cranial irradiation, those who were near pubertal age at the time of transplant, and those who were transplanted for a disease other than acute lymphoblastic leukemia (ALL). During the second year, poor rates of growth were associated only with the use of corticosteroids after transplant.
Treatment of hairy cell leukemia with 2-chlorodeoxyadenosine (2-CdA)
Authors: Estey EH , Kurzrock R , Kantarjian HM , O'Brien SM , McCredie KB , Beran M , Koller C , Keating MJ , Hirsch-Ginsberg C , Huh YO
Published: October 2019

We administered one course of 2-chlorodeoxyadenosine (2CdA) at 4 mg/m2 daily for 7 days by continuous intravenous infusion to 46 patients with hairy cell leukemia. Complete remissions occurred in 36 patients (78%; 95% confidence limits, 63% to 89%), partial remissions in five (11%), and a minor response in one. One patient died of candida sepsis 3 weeks after beginning treatment and three patients were clearly resistant to therapy. These three either had morphologically atypical hairy cells, less than 20% of which expressed Ig light chain on the cell surface, or had failed prior treatment with deoxycoformycin and interferon-alpha. At a median of 37 weeks since discontinuation of therapy, recurrent thrombocytopenia has developed in one patient, whose marrow remains normal, while a bone marrow relapse has occurred in another patient, whose blood counts remain normal. Treatment produced a greater than 50% decrease in neutrophil count in 26 patients, which lasted 3 to 4 weeks and was associated with an increased incidence of febrile episodes. These episodes occurred in 21 patients but were associated with documented infection in only four patients. Decreases in the number of CD4+ lymphocytes appeared to occur regularly after treatment and have persisted for a median of 18 weeks without obvious clinical significance. Although years of follow-up will be needed, our results confirm Piro et al's observation (N Engl J Med 322: 1117, 1990) that 2CdA appears to be highly effective in the treatment of hairy cell leukemia.
Localization of platelet osteonectin at the internal face of the alpha- granule membranes in platelets and megakaryocytes
Authors: Breton-Gorius J , Clezardin P , Guichard J , Debili N , Malaval L , Vainchenker W , Cramer EM , Delmas PD
Published: October 2019

Osteonectin is a 32-Kd phosphoglycoprotein originally described in bone but also found in platelets. Platelet and bone osteonectin are different both structurally and immunologically. We have previously shown that platelet osteonectin, by binding to thrombospondin, is involved in the secretion-dependent phase of the platelet aggregation process. In this study, we used antiosteonectin antibodies in combination with immunogold labeling to investigate by electron microscopy the fine localization of osteonectin within normal and gray platelets. Using both a polyclonal and monoclonal antibody ON3, osteonectin was specifically located at the internal face of alpha- granule membranes within normal platelets. Osteonectin was not distributed within all alpha-granules, probably because of its low platelet content. In addition, using immunofluorescence, osteonectin could also be detected in immature and mature megakaryocytes with a granular pattern of staining, suggesting that osteonectin is synthesized by megakaryocytes. Using platelets from two patients with gray platelet syndrome, osteonectin was absent within all abnormal small alpha-granules, but was detected in some rare normal-sized alpha- granules. In separate double-label studies, thrombospondin and von Willebrand factor did not colocalize with osteonectin in resting platelets. However, osteonectin was located at the inner face of the alpha-granules, as it is for alpha-granule membrane protein GMP-140 and glycoprotein IIb-IIIa. These results, taken together with the fact that monoclonal antibodies to osteonectin bind only to the surface of activated platelets, suggest that platelet osteonectin is redistributed to the cell surface during fusion of alpha-granule membranes with the plasma membrane.
Fatal Transfusion Reaction Due to the Kell Factor
Authors: OTTENSOOSER F. , MELLONE O. , BIANCALANA A.
Published: October 2019

A woman of type B, M, Rh1rh was transfused with blood of the same blood groups, but no direct compatibility tests were made. The patient developed a violent hemolytic reaction, became anuric, and subsequently received four compatible transfusions. The clinical course seemed satisfactory but after a week she died suddenly from a cerebrovascular accident. The donor was Kell positive and the patient Kell negative; the donor’s Kell positive cells had disappeared from her blood one day after the transfusion and her serum contained incomplete Kell antibody.

The patient’s husband and four children were Kell negative, but she had received twenty-three years previously, five transfusions from four donors. Three of these donors were located, two of whom were Kell positive. In tests of the patient’s serum with one hundred and eighty-two random bloods there were fourteen or 7.7 per cent positive reactions, an incidence which is characteristic of anti-Kell in caucasoids.
Case Report: Erythroblastic Anemia
Authors: NIEWEG H. O. , ARENDS A.
Published: October 2019

Atrophy of the spleen was observed in 3 cases of nontropical sprue, 1 case of chronic glomerulonephritis and 1 case of Whipple's disease. The occurrence of orthochromatic normoblasts and Howell-Jolly bodies in the peripheral blood in the absence of splenectomy may be a manifestation of atrophy of the spleen. Folic acid therapy was tried in 1 case and resulted in the disappearance of the orthochromatic normoblasts and fragmentation of the Howell-Jolly bodies.

Some arguments are put forward in favor of the conception that atrophy of the spleen may be caused by a disturbance of cell regeneration due to a chronic folic acid deficiency.
Effects of Total Body X-Irradiation on the Peripheral Blood of the Monkey
Authors: ELDRED EARL , ELDRED BERGLIOT
Published: October 2019

In monkeys subjected to a single dose of LD/50 total body x-irradiation, the periods of initial fall, continued depression and subsequent recovery were determined for total white counts, neutrophils, eosinophils, lymphocytes, monocytes, erythrocytes and platelets.

In general, the effect of irradiation upon the blood picture in the monkey parallels closely that observed in lower animals. Discrepancies in the leukocyte recovery times between human atomic bomb casualties and animals, including the monkey, are noted.

Observations on eosinophils and lymphocytes which bear upon the role of irradiation as a stress stimulus are presented and discussed.
Differences in Antigenic Specificity of Human Normal Adult, Fetal, and Sickle Cell Anemia Hemoglobin
Authors: GOODMAN MORRIS , CAMPBELL DAN H.
Published: October 2019

1. Small differences in the antigenic structures of human normal adult hemoglobin and sickle cell anemia hemoglobin were found to exist. A difference in the serologic behavior of these two hemoglobins attributable to a factor other than that of antigenic specificity was also demonstrated. This factor is thought to pertain to differences in the physicochemical properties of the 2 hemoglobins.

2. Rabbit antiserums failed to show a difference in antigenic specificity, but with chicken antiserums a difference was clearly demonstrated when the in vitro reactions were performed under conditions which produced maximal precipitation. Results suggest that these 2 hemoglobins have a predominance of common antigenic determinants but a small number that are unique for each of the types.

3. Relatively large differences in specificity were found to exist between fetal hemoglobin and adult hemoglobin which suggest that only a few antigenic groups are shared in common by the 2 types.

4. The second, minor hemoglobin in patients with sickle cell anemia was shown to have an antigenic specificity that was either identical with or very similar to that of fetal hemoglobin.
The Nature of Neutral Red Staining Refractive Particles in the Lymphocytes of the Blood of Normal Humans
Authors: HEMPELMANN LOUIS H. , KNOWLTON NORMAN P.
Published: October 2019

1. The refractive neutral red staining particles seen in unstained or supravitally stained lymphocytes have the following properties: They are birefringent and show a polarization cross. Their refractive index is 1.45 to 1.48. They are soluble in fat solvents, in cold alcohol and in strong but not weak acid or base. They stain with lipid stains and basic dyes and are increased in number by the action of neutral red on supravital preparations of blood cells.

2. These properties are interpreted as indicating that the particles seen in human lymphocytes are composed of lipid material. It is probable that much of the fat is of the polar-apolar type, possibly phospholipid in nature.

3. The refractive particles are believed to be the counterparts in human lymphocytes of the phospholipid-containing droplets which occur in somatic cells of other mammalian tissue.

4. The significance of the increase in numbers of these droplets under certain conditions and their relationship to the Golgi apparatus are discussed.
The Use of Nitrogen Mustard in the Management of Two Pregnant Lymphoma Patients
Authors: DEUSCHLE KURT W. , WIGGINS WALTER S.
Published: October 2019

1. It is possible to use nitrogen mustard therapeutically in the management of a patient with "lymphoma" in the second trimester of pregnancy and obtain a viable infant.

2. In view of the findings noted in one of the infants, it is not possible to make a categorical statement with regard to possible deleterious effects of nitrogen mustard on the fetus.
Case Report: Paroxysmal Nocturnal Hemoglobinuria with the Development of Aplastic Anemia
Authors: NELSON M. G. , BRUCE J. H.
Published: October 2019

1. A case of paroxysmal nocturnal hemoglobinuria is presented which developed hemoglobinuria for the first time while in the hospital and showed severe reactions to whole blood transfusion but none to washed red cells.

2. Therapeutic administration of heparin induced severe hemoglobinuric episodes which were followed by a remission in the transfusion reactions to whole blood.

3. Later, aplastic anemia developed with a marked subsidence in the various indicators of the hemolytic process although the erythrocytic defect found in paroxysmal nocturnal hemoglobinuria was still present.
Nonfollicular small B-cell lymphomas: a heterogeneous group of patients with distinct clinical features and outcome
Authors: Berger F , Felman P , Sonet A , Salles G , Bastion Y , Bryon PA , Coiffier B
Published: October 2019

Two hundred sixteen patients with a nonfollicular small cell lymphoma followed up in our department over a 5-year period have been reviewed to define the clinical behavior and survival of patients with each histologic subtype. The respective frequencies of major subtypes were: small lymphocytic/lymphoplasmacytoid lymphoma (immunocytoma, SL/LPL), 28%; large cell-rich immunocytoma (LCRI), 7%; mantle cell lymphoma (MCL), 24%; mucosa-associated lymphoid tissue-lymphoma (MALT-L), 20%; other rare subtypes, 6%; and nonclassified or nonreviewed, 14%. The SL/LPL patients and the MALT-L patients had a relatively indolent disease, usually disseminated for SL/LPL and usually localized for MALT- L. Both subtypes have a long time to treatment failure (TTF; median, 48 and 58 months, respectively) and long survival (median, 118 and 98 months, respectively). The LCRI patients or the MCL patients had more aggressive clinical or biologic features and experienced shorter TTF (median, 26 and 14 months, respectively) and shorter survival (median, 55 and 52 months, respectively). None of these histologic subtypes was associated with a significant cure rate. MALT-L patients did relapse regardless of the initial localization or treatment and at a similar rate to the SL/LPL patients. Factors associated with a worse outcome in nonfollicular small cell lymphoma patients are identical to those described in other lymphoma subtypes: advanced clinical stage, poor performance status, high tumor bulk, and high lactic dehydrogenase or beta 2microglobulin levels. For patients with disseminated disease, standard chemotherapy regimens did not allow a long TTF; therefore, new therapeutic strategies must be developed.
Platelet adhesion to collagen in individuals lacking glycoprotein IV
Authors: McKeown L , Vail M , Williams S , Kramer W , Hansmann K , Gralnick H
Published: October 2019

The Naka isoantigen is expressed on glycoprotein (GP) IV (CD36), a platelet membrane GP that has been identified as having a role in platelet interactions with collagen and thrombospondin and in binding Plasmodium falciparum-infected erythrocytes to endothelial cells and melanoma cells. We have studied normal platelets and Naka- platelets from two Japanese donors that have 1% of GPIV by concentration- dependent antibody binding and flow cytometry. We studied the adherence of normal and Naka- platelets to types I, III, and IV collagen in static and to type I collagen in flowing systems at high shear force. We have also studied aggregation of normal and Naka- platelets to type I collagen. Naka- platelets showed normal or increased aggregation to type I collagen and normal adhesion to types I, III, and IV collagen in the presence of Mg++ or EDTA. Platelet aggregation and adhesion were inhibited by the anti-alpha 2 beta 1 antibody 176D7 to the same extent in Naka- as in normal platelets. We also studied endogenous thrombospondin surface expression and found that thrombin-stimulated Naka- platelets expressed the same amount of thrombospondin as did normal platelets. From our studies with Naka- platelets, we cannot identify a definitive role for GPIV in platelet aggregation, in adhesion to types I, III, and IV collagen, or in endogenous thrombospondin binding to platelets.
Tissue specificity of X-chromosome inactivation patterns
Authors: Gale RE , Wheadon H , Boulos P , Linch DC
Published: October 2019

The analysis of X-chromosome inactivation patterns has been used in a number of clinical situations such as the identification of carrier status in X-linked genetic disorders and the establishment of the monoclonal origin of tumors. Interpretation of the result obtained requires comparison with the constitutive pattern for the individual, and for hematopoietic malignancies, skin biopsies or cultured fibroblasts have often been used as the control tissue because normal cells of the same lineage as the malignancy are not generally available. However, this assumes that patterns in the different tissues are constitutionally the same. We have therefore compared X-chromosome inactivation patterns from peripheral blood (granulocytes, E- cells, and T cells), skin, and muscle from 20 hematologically normal females, and colonic mucosa from 9 individuals. In 11 patients (55%), the results obtained were similar for all tissues of an individual, but in 9 patients, significant differences were observed between tissues. The most consistent feature was a skewing in peripheral blood (> 75% expression of one allele) but not skin and/or muscle. These studies suggest that skin cannot be used as a control tissue for the interpretation of X-chromosome inactivation patterns in hematopoietic cells.
Nonhereditary p53 mutations in T-cell acute lymphoblastic leukemia are associated with the relapse phase
Authors: Hsiao MH , Yu AL , Yeargin J , Ku D , Haas M
Published: October 2019

We have previously reported that greater than 60% of human leukemic T- cell lines possess mutations in the p53 tumor suppressor gene. To determine whether T-cell acute lymphoblastic leukemia (T-ALL) patient samples possess p53 mutations, we screened peripheral blood-and bone marrow-derived leukemia samples, taken at diagnosis and at relapse, for p53 mutations. Exons 4 through 9 and selected intron regions of the p53 gene were analyzed using polymerase chain reaction-single-strand conformation polymorphism and direct sequencing. p53 mutations were found in 0 of 15 T-ALL diagnosis samples, as compared with 10 of 36 (28%) T-ALL relapse samples. To determine whether p53 mutations play a role in the recurrence (relapse) of T-ALL, two special groups of T-ALL patients were studied: (1) a group of 8 relapse patients whose disease was refractory to chemotherapeutic treatment, and (2) a group of 6 “paired” T-ALL cell samples from patients for whom we possess both diagnosis and relapse samples. Three of 8 relapsed patients (37.5%) whose disease was refractory to the reinduction of remission by chemotherapy possessed missense mutations of the p53 gene. All 3 cases had mutations in exon 5. Among the paired samples, 3 of 6 patients harbored p53 mutations at disease recurrence, but possessed only wild- type p53 alleles at diagnosis. One case had mutation on exon 4, 1 case in exon 5, and 1 case in exon 8 with loss of heterozygosity. These data clearly indicate that recurrence of T-ALL is associated with missense mutations in p53. Our results indicate that (1) mutations of p53 do occur in T-ALL in vivo, and such mutations are associated with the relapse phase of the disease; and (2) p53 mutation is involved in the progression of T-ALL. This conclusion is supported by our observation that the introduction of T-ALL-derived mutant p53 expression constructs into T-ALL cell lines further increases their growth rate in culture, enhances cell cloning in methylcellulose, and increases tumor formation in nude mice.
Sustained human hematopoiesis in immunodeficient mice by cotransplantation of marrow stroma expressing human interleukin-3: analysis of gene transduction of long-lived progenitors
Authors: Nolta JA , Hanley MB , Kohn DB
Published: October 2019

We have developed a novel cotransplantation system in which gene- transduced human CD34+ progenitor cells are transplanted into immunodeficient (bnx) mice together with primary human bone marrow (BM) stromal cells engineered to produce human interleukin-3 (IL-3). The IL- 3-secreting stroma produced sustained circulating levels of human IL-3 for at least 4 months in the mice. The IL-3-secreting stroma, but not control stroma, supported human hematopoiesis from the cotransplanted human BM CD34+ progenitors for up to 9 months, such that an average of 6% of the hematopoietic cells removed from the mice were of human origin (human CD45+). Human multilineage progenitors were readily detected as colony-forming units from the mouse marrow over this time period. Retroviral-mediated transfer of the neomycin phosphotransferase gene or a human glucocerebrosidase cDNA into the human CD34+ progenitor cells was performed in vitro before cotransplantation. Human multilineage progenitors were recovered from the marrow of the mice 4 to 9 months later and were shown to contain the transduced genes. Mature human blood cells marked by vector DNA circulated in the murine peripheral blood throughout this time period. This xenograft system will be useful in the study of gene transduction of human hematopoietic stem cells, by tracing the development of individually marked BM stem cells into mature blood cells of different lineages.
Allogeneic bone marrow transplantation with a fixed low number of T cells in the marrow graft [see comments]
Authors: Verdonck LF , Dekker AW , de Gast GC , van Kempen ML , Lokhorst HM , Nieuwenhuis HK
Published: October 2019

Despite prophylaxis with immunosuppressive drugs, severe acute graft- versus-host disease (GVHD) remains a major cause of morbidity and mortality in patients transplanted with unmodified bone marrow (BM) grafts from HLA-identical siblings. Although T-cell depletion of the BM graft has evolved as the most effective method to prevent severe acute GVHD, this beneficial effect is counterbalanced by an increased rate of graft failure and relapse of the disease. To find an approach to T-cell depletion that may avoid these extreme risks, we gave BM recipients a fixed low number of 1 x 10(5) donor T cells per kilogram of recipient's body weight in the graft. This corresponds with 99% T-cell depletion and is achieved by the addition of T cells to the graft that was previously depleted of T cells. A total of 70 patients with hematologic malignancies or aplastic anemia, including 40 patients with standard- risk leukemias, received BM grafts, depleted of T cells according to this approach, from HLA-identical siblings. The preparative regimen consisted of cyclophosphamide and total body irradiation. The patients also received a short course of cyclosporine posttransplant. Graft failure did not occur. Acute GVHD, only grade I or II, was seen in 70% of the patients and was limited to the skin in all patients. Chronic GVHD occurred in 31% of the patients and, with the exception of 1 patient, was limited to the skin as well. Relapse occurred in 3 of 40 (8%) patients with standard-risk leukemias, resulting in a projected survival at 5 years of 80%. Patients with standard-risk diseases had a procedure-related mortality of 11%. Quality of life, determined 1 year after BM transplant, was good in almost all patients with standard-risk diseases. Thus, this approach of T-cell depletion may be an approach that avoids the development of severe acute and chronic GVHD without damaging the function or antileukemic effect of the graft and that has a low transplant-related morbidity and mortality.
Human MafG Is a Functional Partner for p45 NF-E2 in Activating Globin Gene Expression
Authors: Blank Volker , Kim Min J. , Andrews Nancy C.
Published: October 2019

Mammalian globin gene expression is activated through NF-E2 elements recognized by basic-leucine zipper proteins of the AP-1 superfamily. The specificity of NF-E2 DNA binding is determined by several nucleotides adjacent to a core AP-1 motif, comprising a recognition site for transcription factors of the Maf subfamily. Earlier work proposed that p18(MafK) forms a heterodimer with hematopoietic-specific protein p45 NF-E2 to activate transcription through NF-E2 sites. However, there was no direct evidence that p18(MafK) serves this function in vivo; in fact, mice lacking p18(MafK) have no phenotype. Here we describe a novel cDNA clone that encodes the human homolog of chicken MafG. Human MafG heterodimerizes with p45 NF-E2 and binds DNA with specificity identical to that of purified NF-E2 DNA binding activity. A tethered heterodimer of p45 and MafG is fully functional in supporting expression of α- and β-globin, and in promoting erythroid differentiation in CB3, a p45-deficient mouse erythroleukemia cell line. These results indicate that human MafG can serve as a functional partner for p45 NF-E2, and suggest that the p45/MafG heterodimer plays a role in the regulation of erythropoiesis.
Immunotherapy With Recombinant Human Interleukin-2 and Recombinant Interferon-α in Lymphoma Patients Postautologous Marrow or Stem Cell Transplantation
Authors: Nagler A. , Ackerstein A. , Or R. , Naparstek E. , Slavin S.
Published: October 2019

Immune-mediated effects appear to play a major role in controlling minimal residual disease (MRD). We, therefore, investigated the role of recombinant human interleukin-2 (rIL-2) given concomitantly with interferon-α (IFN-α) in malignant lymphoma (ML) patients with responding disease following autologous bone marrow or blood stem cell transplantation (ABSCT). Fifty-six patients were included in this investigation. Thirty-two patients had non-Hodgkin's lymphoma (NHL) and 24 patients had Hodgkin's disease (HD). Sixty-one patients (NHL 36, HD 25) served as historical controls. Patients from both groups had similar demographic characteristics, the same stage of disease at presentation, status of disease at transplantation, conditioning regimens, and type of transplant. rIL-2 and IFN-α were selfadministered in two cycles beginning 2.5 to 10.5 months (median, 4 months) posttransplant and separated by a 4-week interval. Each cycle consisted of IFN-α subcutaneously (SC) 3 × 106 U/d × 5 d/wk combined with rIL-2 SC 3 to 6 IU/m2/d × 5 d/wk for 4 weeks. The incidence of survival and disease-free survival (DFS) was significantly higher in the group under investigation than in the historical controls (P < .01). Of 56 patients with ML treated with IFN-α + rIL-2, 45 patients are DFS (80.4%) after a follow-up of 7 to 78 months (median, 34 months), whereas in the historical controls, 32 of 61 (52.5%) patients are disease free, in a follow-up of 4 to 84 months (median, 23 months) posttransplant (P < .01). Our preliminary results are encouraging and suggest that home administered immunotherapy with IFN-α and rIL-2 is relatively well tolerated and may intensify remission in ML patients with MRD following ABSCT.
Prognostic Significance of Fluorescence Intensity of Surface Marker Expression in Childhood B-Precursor Acute Lymphoblastic Leukemia. A Pediatric Oncology Group Study
Authors: Borowitz Michael J. , Shuster Jonathan , Carroll Andrew J. , Nash Michael , Look A. Thomas , Camitta Bruce , Mahoney Donald , Lauer Stephen J. , Pullen D. Jeanette
Published: October 2019

This report describes the prognostic significance of the intensity of surface membrane antigen expression in a series of 1,231 children older than 1 year with newly diagnosed B-precursor acute lymphoblastic leukemia (ALL) treated on Pediatric Oncology Group (POG) treatment protocols. All patients had dual-color flow cytometric immunophenotyping performed at a central reference laboratory with a standard panel of monoclonal antibodies. The flow cytometers used in the study were calibrated with a standard fluorescence microparticle that permitted conversion of relative fluorescence channels to standard units of mean equivalents of soluble fluorochrome (MESF). In univariate analysis, fluorescence intensity of CD45 and CD20 was significantly associated with event-free survival (EFS), whereas other markers showed no significant correlation with outcome. Patients whose blasts were greater than the 75th percentile of intensity for CD45 (corresponding to 18,000 MESF units with CD45-FITC, or about 8% of the intensity of normal lymphocytes) fared significantly worse than those with lower-density CD45, and those whose blasts were greater than the 25th percentile of intensity for CD20 (corresponding to 17,900 MESF units with CD20-PE) had a poorer EFS. The intensity of both CD45 and CD20 was independently correlated with outcome. There was no significant correlation between intensity of expression of either antigen and traditional clinical risk factors, ploidy, or t(9; 22) or t(1; 19). All patients with t(4; 11) had CD45 intensity greater than the 75th percentile, but CD45 intensity retained its prognostic significance after adjusting for t(4; 11). In multivariate analysis, both CD45 intensity greater than the 75th percentile and CD20 intensity greater than the 25th percentile were significantly correlated with poor outcome independently of previously reported poor prognostic factors including National Cancer Institute (NCI) risk group, ploidy, trisomies of 4 and 10, and adverse translocations including t(1; 19), t(9; 22), and t(4; 11). We conclude that in childhood B-precursor ALL, the intensity of expression of CD20 and CD45 provides prognostic information not available from simple consideration of antigen expression as positive or negative, and adds to that obtained from traditional clinical and biologic risk factors.
Rapid Engraftment Without Significant Graft-Versus-Host Disease After Allogeneic Transplantation of CD34+ Selected Cells From Peripheral Blood
Authors: Urbano-Ispizua Alvaro , Rozman Ciril , Martı́nez Carmen , Marı́n Pedro , Briones Javier , Rovira Montserrat , Féliz Pedro , Cruz Viguria Maria , Merino Ana , Sierra Jorge , Mazzara Roberto , Carreras Enric , Montserrat Emilio
Published: October 2019

We have prospectively evaluated the feasibility and results of the biotin-avidin immunoadsorption method (Ceprate SC system) for a phase I/II study of T-cell depletion of granulocyte colony-stimulating factor (G-CSF ) mobilized peripheral blood progenitor cells (PBPC) for allogeneic transplantation. Twenty consecutive patients, median age, 40 years (21 to 54) and diagnoses of chronic myeloid leukemia in chronic phase (n = 5), acute myeloblastic leukemia (n = 7), acute lymphoblastic leukemia (n = 2), chronic myelomonocytic leukemia (n = 1), refractory anemia with excess of blasts in transformation (n = 3), histiocytosis X (n = 1), and chronic lymphocytic leukemia (n = 1), were conditioned with cyclophosphamide (120 mg/kg) and total body irradiation (13 Gy; 4 fractions). HLA identical sibling donors received G-CSF at 10 μg/kg/d subcutaneously (SC); on days 5 and 6 (19 cases) and days 5 to 8 (1 case) donors underwent 10 L leukapheresis. PBPC were purified by positive selection of CD34+ cells using immunoadsorption biotin-avidin method (Ceprate SC) and were infused in the patients as the sole source of progenitor cells. No growth factors were administered posttransplant. The median recovery of CD34+ cells after the procedure was of 65%. The median number of CD34+ cells infused in the patients was 2.9 (range, 1.5 to 8.6) × 106/kg. The median number of CD3+ cells administered was 0.42 × 106/kg (range, 0.1 to 2). All patients engrafted. Neutrophil counts <500 and <1,000/μL were achieved at a median of 14 days (range, 10 to 18) and 15 days (range, 11 to 27), respectively. Likewise, platelet counts <20,000 and <50,000/μL were observed at a median of 10 days (range, 6 to 23) and 17 days (range, 12 to 130), respectively. Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine plus methylprednisolone. No patient developed either grade II to IV acute or extensive chronic GVHD. After a median follow-up of 7.5 months (range, 2 to 22) three patients have relapsed, and one of them is again in hematologic and cytogenetic remission after infusion of the donor lymphocytes. Two patients died in remission: one on day +109 of pulmonary aspergillosis and the other on day +251 of metastasic relapse of a previous breast cancer. Sixteen of the 20 patients are alive in remission after a median follow-up of 7.5 months (range, 2 to 22). In conclusion, despite the small number of patients and limited follow-up, it appears that this method allows a high CD34+ cell recovery from G-CSF mobilized PBPC and is associated with rapid engraftment without significant GVHD, and with low transplant related mortality.