bluuebunny/crossref_metadata_2025 huggingface dataset:
Some Interrelationships of Blood and the Fava Bean Principle in Vitro
Authors: CREGER WILLIAM P. , GIFFORD HOUGHTON
Published: October 2019

1. Saline suspensions of human red cells, as well as those of several animal species, were agglutinated by normal saline extracts of the Fava bean.

2. This agglutination was potentiated in titer 100-fold in a medium of 10 per cent acacia, as a diluent.

3. The inhibition of the hemagglutination action of the Fava bean extract by human serum was apparently attributable to the gamma globulin fraction.

4. The Fava bean principle could be transferred from cell to cell, as shown by heat-elution and acacia technics.

5. Fava-sensitized red cells did not exhibit increased susceptibility in the test tube to complement, hemolysin, or osmotic or mechanical fragility.

6. The mechanism of in vivo red cell destruction in Favism is as yet unknown, but a special immunologic susceptibility to the action of the bean’s principle is suspected in certain persons.

7. It is suggested that the relation of acacia to Fava-sensitized red cells may form the basis of a diagnostic test for Favism in the early, acute stages of the disease.
Methods: Non-Wettable Surfaces
Authors: TULLIS JAMES L. , ROCHOW EUGENE G.
Published: October 2019

Types of non-wettable surfaces and their means of application are outlined and discussed.
Red Cell Thickness in Normals and in Pernicious Anemia
Authors: LARSEN GERHARD
Published: October 2019

A method previously described by Williams and Wyckoff, Bessis, and others is adapted for measuring the thickness of the dried blood cell. Statistical analysis of the results reveals the presence in pernicious anemia of a distinct type of cell which in the dried blood film has a mean diameter 1 µ above normal and a thickness 0.224 µ less than normal. These pathologic cells, which thus flatten out when spread on a glass slide, also shrink more than do normal cells when dried in a blood film.
Surface antigenic profile and globin phenotype of two new human erythroleukemia lines: characterization and interpretations
Authors: Papayannopoulou T , Nakamoto B , Kurachi S , Tweeddale M , Messner H
Published: October 2019

Detailed characterization of the composite phenotype of two newly established erythroleukemia lines (OCIM1, OCIM2) shows that these lines share many of their erythroid markers (ie, surface antigens and globin program) as well as several of their nonerythroid properties (myeloid/monocytic/megakaryocytic) with the two known erythroleukemia lines (K562, HEL). In addition, each displays novel and instructive features. We argue that the surface and globin phenotype of all erythroleukemia lines is nonrandom and that it may be of physiologic relevance; it could represent the most prevalent phenotype of cells transformed by leukemia in vivo, and it raises the possibility that cells with similar potentials exist transiently during normal hematopoietic differentiation before their irreversible commitment to a single lineage. As such, these cells demonstrate a greater phenotypic adaptability in vitro than do their single lineage-committed counterparts since they can differentiate toward more than one lineage.
Neutrophil and monocyte adherence to and migration across monolayers of cytokine-activated endothelial cells: the contribution of CD18, ELAM-1, and VLA-4
Authors: Hakkert BC , Kuijpers TW , Leeuwenberg JF , van Mourik JA , Roos D
Published: October 2019

Pretreatment of endothelial cells with cytokines enhances the adherence of leukocytes, a process that is mediated by surface proteins expressed on both cell types. A three-dimensional model system for the simultaneous determination of leukocyte adherence and migration was used to study the contribution of CD11/CD18, endothelial leukocyte- adhesion molecule-1 (ELAM-1) and VLA-4 in neutrophil and monocyte adherence to and migration through cytokine-activated endothelial cells. Pretreatment of endothelial cells for 4 hours with recombinant interleukin-1 beta (rIL-1 beta) was found to enhance neutrophil adherence and migration to a much greater extent than monocyte adherence and migration. Neutrophil adherence was almost completely prevented by the combined use of monoclonal antibodies (MoAbs) against ELAM-1 and CD18. Although ELAM-1 has been designated an endothelial cell-specific cytokine-inducible receptor for neutrophils, we observed that ENA2, an anti-ELAM-1 MoAb, significantly reduced monocyte adherence about 30%. MoAbs against VLA-4, the ligand of the cytokine- inducible receptor VCAM-1, did not affect monocyte adherence. However, the combined use of the MoAbs against CD18, ELAM-1, and VLA-4 had a very strong and additive inhibitory effect on rIL-1 beta-induced monocyte adherence. The anti-CD18 MoAb reduced both rIL-1 beta-induced neutrophil and monocyte migration far below the level of the unstimulated controls, whereas neither the anti-ELAM-1 nor the anti-VLA- 4 MoAb significantly affected the process of migration. Our results indicate that neutrophils and monocytes initially adhere to cytokine- activated endothelial cells by CD18-independent and (to a lesser extent) by CD18-dependent mechanisms and subsequently change gears to a completely CD18-dependent migratory mechanism.
High expression of transforming growth factor-beta long cell cycle times and a unique clustering of S-phase cells in patients with acute promyelocytic leukemia [see comments]
Authors: Raza A , Yousuf N , Abbas A , Umerani A , Mehdi A , Bokhari SA , Sheikh Y , Qadir K , Freeman J , Masterson M
Published: October 2019

Expression of transforming growth factor-beta (TGF-beta), which inhibits the proliferation of hematopoietic progenitors, was investigated simultaneously with cell cycle characteristics in 63 bone marrow biopsies from 23 cases with acute promyelocytic leukemia (APL). Bromodeoxyuridine (BrdU) was administered to every patient (17 newly diagnosed) for determination of the labeling index (LI) and the durations of S-phase (Ts) and the cell cycle (Tc) of leukemic promyelocytes. APL cases had lower LI both in the bone marrow aspirate (6.1% v 11.4%, P = .008) and biopsy (21.1% v 28.0%, P = .001) and longer Tc (93.6 hours v 56.0 hours, P = .002) when compared with other French-American-British subtypes. TGF-beta expression (detected by a monoclonal anti-TGF-beta 2/beta 3 antibody) was dramatically high, especially in interstitial areas of the biopsies. S-phase cells were found as geographically restricted islands of proliferation (GRIPs) in 20 of 22 cases. Weekly biopsies showed an increment in TGF-beta on day 7 of therapy in 13 of 17 cases, while in vivo differentiation was noted in 9 of 15. We conclude that the presence of high TGF-beta expression may explain the biologic basis for the slowly cycling nature of leukemic promyelocytes in APL as well as the unique clustering of S- phase cells observed in GRIPs.
Oxidative red blood cell membrane injury in the pathophysiology of severe mouse beta-thalassemia
Authors: Advani R , Rubin E , Mohandas N , Schrier SL
Published: October 2019

In severe human beta-thalassemia, the pathophysiology relates to accumulation of excess alpha-globin chains at the membrane. One hypothesis is that membrane-associated alpha-globin by virtue of it's iron or hemichromes produces oxidation of adjacent membrane proteins. The availability of a mouse model of severe beta-thalassemia, as well as a transgenic (thalassemic-sickle) mouse that expresses 12% of human beta s-chain, has allowed us to study the effect of graded accumulation of alpha-chains at the red blood cell (RBC) membrane on the clinical status of the animal and on the material properties of its RBCs. Proteins from control, beta-thalassemic, and transgenic mouse RBC membranes were analyzed for evidence of oxidation, as measured by thiol- disulfide exchange chromatography, which detects intramolecular sulfhydryl oxidation. Ratios of oxidized globin to protein 7 were calculated and increased amounts were seen in thalassemic mice as compared with control mice and transgenic mice. Furthermore, there were increased amounts of thiol-free protein 4.1 in the thalassemic mice, compared with very small amounts in the control mice and intermediate amounts in the transgenic mice. Membrane mechanical stability as assessed by ektacytometry showed that the thalassemic mouse RBCs were markedly unstable. Transgenic mouse RBCs showed intermediate levels of membrane instability compared with the controls. We propose that this oxidized globin, in conjunction with oxidized protein 4.1, accounts (at least in part) for membrane instability. A 12% increase in beta s- globin chain synthesis (by decreasing excess globin available) confers considerable protection against both oxidative damage and the consequent membrane instability.
Spectrum of toxicities of amino acid methyl esters for myeloid cells is determined by distinct metabolic pathways
Authors: Thiele DL , Lipsky PE
Published: October 2019

L-leucine methyl ester (Leu-OMe), Leu-Leu-OMe, Phe-OMe, and Glu-(OMe)2 are toxic to mononuclear phagocytes (M phi) and neutrophils. In the present studies, the mechanism of this toxicity was examined. A concentration of NH4Cl known to neutralize lysosomal pH and to block conversion of Leu-OMe to the dipeptide condensation product Leu-Leu-OMe inhibited Leu-OMe- or Glu-(OMe)2- but not Leu-Leu-OMe-mediated M phi toxicity. Leu-OMe-, Glu-(OMe)2-, or Leu-Leu-OMe-mediated killing of M phi was prevented by Gly-Phe-CHN2, a specific inhibitor of the thiol protease, dipeptidyl peptidase I (DPPI). Neither NH4Cl nor Gly-Phe-CHN2 prevented Phe-OMe-mediated M phi toxicity. In contrast, inhibition of M phi serine esterase activity prevented Phe-OMe- but not Leu-OMe- or Glu- (OMe)2-mediated killing of M phi. The myeloid tumor lines U937, HL60, and THP-1 were found to be uniformly enriched in DPPI and susceptible to Leu-Leu-OMe but not Leu-OMe toxicity. Whereas HL60 were resistant to Phe-OMe, THP-1 cells were killed by this agent. Incubation of peripheral blood mononuclear cells with Leu-OMe resulted in loss of natural killer (NK) functions and cytotoxic T lymphocytes (CTL) precursors, a process that requires the DPPI-dependent generation of membranolytic polymerization products. Phe-OMe had no toxic effects on NK cells or CTL precursors. These results indicate that Leu-OMe and Glu- (OMe)2 toxicity for M phi is related to the production of higher molecular weight hydrophobic polymerization products via the sequential action of two nonserine esterase lysosomal enzymes. In contrast, Phe- OMe toxicity for myeloid cells was found to correlate with serine esterase-mediated intracellular trapping of high concentrations of the free amino acid Phe. These distinct enzymatic mechanisms may provide a unique means of targeting agents capable of selectively deleting cells of myeloid lineage.
The Demonstration of the "L.E." Phenomenon in Patients with Penicillin Hypersensitivity
Authors: WALSH JOHN R. , ZIMMERMAN HYMAN J.
Published: October 2019

Of 6 patients with penicillin reactions, 3 whose reactions were severe were found to possess the "L. E." factor in the plasma and "L. E." cells in the bone marrow. The 3 who failed to show the "L. E". phenomenon had only minimal reactions to penicillin. Since the group studied was small, no conclusions regarding the incidence of the "L. E." phenomenon in penicillin reactions can be drawn. The findings are discussed in the light of the possibility that the phenomenon may be related to hypersensitivity reactions as well as to systemic lupus erythematosus.
Quantitative Studies of the Influence of Plasma Proteins and Hematocrit on the Erythrocyte Sedimentation Rate
Authors: MEYERS ALDULA J. , TREVORROW VIRGINIA , WASHBURN ALFRED H. , MUGRAGE EDWARD R.
Published: October 2019

The failure to devise a satisfactory means of adjusting all sedimentation rates to a standard cell volume, together with a critical analysis of the results of other investigators, casts serious doubt on the validity of any such correction charts.

Two formulae are presented by means of which the sedimentation rate of a given sample of blood can be predicted on the basis of the plasma/cell ratio and plasma proteins concentrations. One formula utilizes the results of saltingout analysis of plasma proteins, the other those of electrophoretic analysis of serum proteins. The most influential factors in determininsg sedimentation rate are plasma/cell ratio, fibrinogen, alpha-2 globulin, and gamma globulin. The results suggest that the effective concentrations of the plasma proteins are most adequately expressed as concentrations per unit of cell volume.

Addition of purified protein fractions (fibrinogen, gamma globulin, and albumin) to the blood in vitro confirm the findings from the statistical studies and suggest a direct cause and effect relationship between the concentration of these proteins and the rate of erythrocyte sedimentation.

Determination of the erythrocyte sedimentation rate cannot give, per se, any clue as to the level of any one of the responsible factors. An appreciation of the multiple factors involved is essential for an adequate interpretation of the clinical significance of a sedimentation rate determination.
Thrombocytopathic Purpura with Normal Prothrombin Consumption
Authors: DE VRIES ANDRÉ , SHAFRIR ELEAZAR , EFRATI PINCHAS , SHAMIR ZVI
Published: October 2019

Three cases of thrombocytopathic purpura with partial platelet dysfunction are reported. The platelets were defective in clot retraction, but normal in prothrombin conversion.
Clinical Evaluation of a New Antimetabolite, 6-Mercaptopurine, in the Treatment of Leukemia and Allied Diseases
Authors: BURCHENAL J. H. , MURPHY M. L. , ELLISON R. R. , SYKES M. P. , TAN T. C. , LEONE L. A. , KARNOF-SKY D. A. , CRAVER L. F. , DARGEON H. W. , RHOADS C. P.
Published: October 2019

1. A new antimetabolite, 6-mercaptopurine, has been shown to produce good clinical and hematologic remissions in fifteen out of forty-five children with acute leukemia. Another ten showed partial remissions and clinical improvement.

2. Remissions in adults with acute leukemia have occasionally been brought about by 6-mercaptopurine, and in a few cases it has produced temporary remissions in both the early and the late stages of chronic myelocytic leukemia.

3. The compound has been effective in some children whose disease was resistant to the folic acid antagonists, as shown by the fact that out of twenty-four children with acute leukemia whose disease had been proved to be resistant to amethopterin, five had good clinical and hematologic remissions and five had partial remissions with some marrow and clinical improvement. Some benefit was seen in eight out of eighteen patients whose disease was resistant to ACTH and cortisone.

4. In children the daily oral administration of 2.5 mg./Kg. rarely caused toxic manifestations, but continued therapy at this dose in adults or at higher levels in children occasionally produced bone marrow depression or gastrointestinal symptoms.

5. There is evidence that the therapeutic resistance of the acute leukemias to 6MP develops somewhat more rapidly than it does to the folic acid antagonists but there is, as yet, no laboratory or clinical evidence of cross resistance between these two types of antimetabolites.

6. In a total of thirty-five patients with lymphomas and miscellaneous carcinomas and sarcomas, 6MP did not produce any definite clinical improvement at doses which produced hematologic toxicity.

7. Although 6-mercaptopurine acts as a purine antagonist in certain forms of bacteria, the exact mechanism of its action in leukemia is at present unknown. Since its mode of action appears to differ from that of other agents previously employed clinically in the treatment of leukemia, this compound would appear to be of fundamental as well as practical interest.
Hyperglobulinemia as the Cause of Hemophilia-like Disease
Authors: NILSSON INGA M. , WENCKERT ANDERS
Published: October 2019

A case is reported of a probably familial hyperglobulinemia in a 44 year old woman with a circulating anticoagulant, antagonistic to the initial phase of the coagulation, most likely to the antihemophilic globulin. The anticoagulant factor was separated by means of salt precipitation and by electrophoresis. It was shown to be identical with one of the patient's pathologic gamma globulins. There is no reason to suppose an isoimmunization process. The coagulation defect is therefore assumed to be a secondary phenomenon to the patient's probably familial protein disturbance. Dysproteinemia has been observed in hemophilia-like diseases several times but the etiologic connection between the coagulation anomaly and the protein disease has not been previously established. On the other hand, even serious dysproteinemias seem rarely to be complicated by a circulating anticoagulant.

A connection between the sedimentation rate and the coagulation of the blood was further shown to be present in our patient.
Plasma Thromboplastin Component (PTC) A Hitherto Unrecognized Blood Coagulation Factor Case Report of PTC Deficiency
Authors: WHITE SIDNEY G. , AGGELER PAUL M. , GLENDENING MARY BETH
Published: October 2019

1. A new hemorrhagic disease characterized by a prolonged whole blood coagulation time caused by the delayed formation of thrombin is described.

2. The patient suffering from this disease was found to have normal concentrations of all previously described coagulation factors.

3. Because the coagulation defect could be completely corrected by the addition of tissue thromboplastin or platelet-poor hemophilic plasma and because tests for prothrombin and prothrombin conversion accelerators gave normal results and prothrombin utilization was impaired, it was concluded that the missing factor was concerned with the production of thromboplastin in the plasma. Accordingly it was named the plasma thromboplastin component (PTC).

4. The clinical and laboratory resemblance of PTC deficiency and hemophilia is striking. However, the two diseases can be distinguished by the fact that normal serum, the 45 to 50 per cent saturated ammonium sulfate fraction of normal plasma, the citrate eluate from barium sulfate, which has been mixed with normal plasma or serum, and hemophilic plasma correct PTC deficiency but are ineffective in hemophilia. On the other hand, while the 33 per cent saturated ammonium sulfate fraction of normal plasma, barium sulfate adsorbed normal plasma, Cohn's Fraction I and PTC-deficient plasma correct the hemophilic defect, they are ineffective in PTC deficiency.

5. A method for the partial purification and concentration of the PTC factor is described.
Malignant Lymphomas Their Classification and Relation to Leukemia
Authors: BERMAN LAWRENCE
Published: October 2019

The value of complete hematologic study, including examination of the blood and bone marrow and imprints as well as sections of lymph nodes, for the diagnosis of lymphoblastomas has been indicated. The various forms of these tumors have been described and defined, and their relationships to each other and to leukemia have been discussed and illustrated. A simple working classification has been presented.
Binding of Acid and Basic Dye at Varied pH by Blood and Bone Marrow Cells of Man
Authors: WEISS LEON PAUL
Published: October 2019

In acid solution, tissues tend to acquire a net positive charge and bind acid dye; binding of basic dye is enhanced in basic solution where tissues tend to possess a net negative charge. A measure of the acid or basic strength of a given tissue structure is afforded by its ability to bind basic or acid dye over a range of pH.

The capacity of blood cell components to bind light green and methylene blue in the pH range 0.05 to 12.3, and their affinity for eosin after acetylation and treatment with nitrous acid, procedures used to block or destroy primary and secondary amines and guanidino groups, was studied. Observations were also made of blood smears stained with the Serra test for arginine.

Erythrocytes bound light green through pH 12.3 and methylene blue to pH 6.0. The ground substance of white blood cells and immature blood cells stained with light green through about pH 9 consistently, and often to pH 11: methylene blue was bound through approximately pH 4.0. Eosinophilic granules bound light green through pH 12.0, and did not bind methylene blue. Basophilic granules bound methylene blue in the pH range 8.0 to 1.0 but were dissolved in more acid and basic solutions: they bound no acid dye. Neutrophilic granules were stained with methylene blue through pH 5.3. Nuclei of immature blood cells and monocytes bound methylene blue to about pH 4.0: polymorphous nuclei and nuclei of lymphocytes and normoblasts were stained very faintly to pH 0.05. Many nuclei bound light green to about pH 12.0.

Nuclear and cytoplasmic acidophilia was reduced or destroyed by acetylation or treatment of smears with nitrous acid. Nuclei and cytoplasm of blood cells were stained with the Serra test. Eosinophilic granules were stained strongly.

The cytochemical significance of certain experimental findings have been discussed.
Studies on the Anticoagulant Phenylindanedione
Authors: GUTTAS CHARLES G. , MOLONEY WILLIAM C. , SISE HERBERT S.
Published: October 2019

1. Phenylindanedione-induced hypoprothrombinemia is rapidly counteracted by a vitamin K1 emulsion. This hypoprothrombinemia is antagonized at a more rapid rate than that induced by any of the other commonly employed prothrombopenic agents.

2. The optimum dose of the vitamin K1 emulsion in emergency is 5 mg. per Kg. Since a refractoriness to the prothrombopenic action of phenylindanedione ensues, smaller doses should be used in situations where there is to emergency.
Action of Alpha-Tocopherol Phosphate on Hemolysis in Paroxysmal Nocturnal Hemoglobinuria
Authors: WILLIAMS MARJORIE J. , CLAPP MARY P.
Published: October 2019

1. The action of alpha-tocopherol phosphate (αTPO4), an anticoagulant, on PNH hemolysis was studied by both in vitro and in vivo means. It was found that the addition of αTPO4 to the modified Ham test inhibited lysis of PNH erythrocytes. During the intramuscular administration of αTPO4, the susceptibility of the erythrocytes to lysis by fresh acidified serum progressively decreased. These cells, after incubation with the serum, gave a positive Coombs test.

2. The possible significance of these findings is briefly discussed.

3. It is concluded that αTPO4 has no therapeutic value in PNH.
Studies on the Metabolism of Fibrinogen in Two Patients with Congenital Afibrinogenemia
Authors: GITLIN DAVID , BORGES WAYNE H.
Published: October 2019

1. The rate of disappearance from the plasma of intravenously administered fibrinogen has been studied immunochemically in two children with congenital afibrinogenemia.

2. About half of the administered fibrinogen disappeared from the circulation in the first forty-eight hours, thus emphasizing the importance of the extravascular pool of plasma protein.

3. After the first two days, the fibrinogen followed a logarithmic decay curve, with a half life of four days; this is so close to the half life of 5.6 days estimated from radioisotope turnover studies that it indicates that the fundamental defect in these patients is a failure to synthesize adequate quantities of fibrinogen.

4. Traces of fibrinogen were detected immunochemically in the plasma of each patient; in one of them, the small amount found could not readily be attributed to prior infusion of blood or blood products.

5. The connective tissue in biopsy specimens of skin and muscle from one patient, when examined with fluorescein-labelled antisera, was found to be specifically deficient in fibrinogen. This deficit was quickly rectified by intravenous administration of fibrinogen.
Age, Sex, and Race Selection at Death from Leukemia and the Lymphomas
Authors: GILLIAM ALEXANDER G.
Published: October 2019

Attention has been called to the distinction between "age incidence", which is a measure of risk, and "age distribution" which is not such a measure except under certain unusual circumstances which probably do not exist for any hospital experience in the United States. Examples to illustrate this distinction were drawn from death data for deaths attributed to leukemia and the lymphomas in the United States in 1949.

The sex and race selection have been recorded for the types of leukemia and lymphoma separable in the sixth revision of The International List of Causes of Death. The age selection at death attributed to the numerically important of these causes has also been presented.

To determine the age, sex, and race selection (incidence) of these diseases, with full confidence in adequacy of their classification, will require a cooperative study designed to apply uniform diagnostic technics to all cases occurring in some definable population such as a large city or a state. Data derived from individual hospitals or from literature summations are generally inadequate for this purpose.
Be, a New "Private" Blood Factor
Authors: DAVIDSOHN ISRAEL , STERN KURT , STRAUSER EMORY R. , SPURRIER WILMA
Published: October 2019

A new "private" blood factor, designated as Berrens, was demonstrated with the aid of an immune isoantibody which was found in a woman sensitized by previous transfusion of her husband's blood and by pregnancy. The antibody had a relatively high titer and produced hemolytic disease of the newborn in the second child, manifested as jaundice and anemia. The factor was present in four out of seven members of three generations: grandmother, husband, his sister, and second child. It was absent in the father of the husband, his wife, the first child, and in four hundred and forty-eight random bloods.

Husbands should not be used as donors of blood for their wives.

Tests for immune isoantibodies developing in pregnant women are incomplete unless red cells of the child and/or father are included as tests cells.
Cloning of the human homologue of the murine flt3 ligand: a growth factor for early hematopoietic progenitor cells
Authors: Lyman SD , James L , Johnson L , Brasel K , de Vries P , Escobar SS , Downey H , Splett RR , Beckmann MP , McKenna HJ
Published: October 2019

Using a fragment of the murine flt3 ligand as a probe, we have succeeded in cloning a human flt3 ligand from a human T-cell lambda gt10 cDNA library. The human and murine ligands are 72% identical at the amino acid level. Analysis of multiple cDNA clones shows that alternative splicing of the human flt3 mRNA can occur at a number of positions. A recombinant soluble form of the human flt3 ligand stimulates the proliferation and colony formation of a subpopulation of human bone marrow cells that are CD34+ and are enriched for primitive hematopoietic cells. In addition, the human flt3 ligand also stimulates the proliferation of cells expressing murine flt3 receptors. Northern blot analysis shows widespread expression of flt3 ligand mRNA transcripts in human tissues.
A randomized study of erythropoietin and granulocyte colony-stimulating factor (G-CSF) versus placebo and G-CSF for patients with Hodgkin's and non-Hodgkin's lymphoma undergoing autologous bone marrow transplantation
Authors: Chao NJ , Schriber JR , Long GD , Negrin RS , Catolico M , Brown BW , Miller LL , Blume KG
Published: October 2019

Anemia is a universal finding in patients undergoing autologous bone marrow transplantation (BMT). Effective therapies to increase the number of autologous red blood cells could result in a lower morbidity and mortality associated with red blood cell transfusions. We examined whether the addition of erythropoietin (Epo) to intensive therapy supported by progenitor cell transplantation and granulocyte colony- stimulating factor (G-CSF) would result in a lower requirement for red blood cell transfusions. Thirty-five patients with lymphoma were randomized to receive Epo versus placebo. Epo (600 U/kg three times per week) or placebo was begun 3 weeks before administration of high-dose therapy. Epo was held during the week of the preparatory regimen, and restarted on the day after BMT. All patients also received G-CSF following BMT. No significant differences were noted between the two groups in terms of patient characteristics at pretreatment or post-BMT evaluation. There were no differences in the total number of red blood cell units transfused (median Epo: 8 v placebo: 6, P = .22) nor the number of platelet transfusions given (median Epo: 12 v placebo 5, P = .14). Engraftment of granulocytes (absolute neutrophil count > or = 500/microL) occurred in a median of 12 days (range, 9 to 33) for the patients receiving Epo and G-CSF, compared with a median of 10 days (range, 8 to 22) for those receiving placebo and G-CSF (P = .70). Likewise, there were no differences in the time to platelet count > or = 20,000/microL without further transfusions with a median of 22 days (range, 15 to 150+) for those receiving Epo and G-CSF compared with a median of 20 days (range, 11 to 54) for those patients receiving placebo and G-CSF (P = .28). The combination of G-CSF and Epo as administered in this study appears to be safe but does not result in an improvement in the total number of red blood cell transfusions or total number of single donor platelet units transfused.
Differential regulation of stem cell factor mRNA expression in human endothelial cells by bacterial pathogens: an in vitro model of inflammation
Authors: Koenig A , Yakisan E , Reuter M , Huang M , Sykora KW , Corbacioglu S , Welte K
Published: October 2019

Production of hematopoietic growth factors by endothelial cells plays a pivotal role during inflammatory processes. Stem cell factor (SCF) is known to be expressed constitutively in endothelial cells. To investigate the regulation of this cytokine expression by inflammatory stimuli, we cocultured human umbilical vein endothelial cells (HUVEC) with various gram-negative bacterial strains (Escherichia coli, Yersinia enterocolitica, Chlamydia trachomatis, and Neisseria meningitidis, respectively). Experiments were performed with bacterial concentrations ranging from 10(2) to 10(7) bacteria/mL for 3 hours. SCF- specific mRNA expression was studied using Northern blot analysis. Stimulation with the enteropathogenic bacterial strains Y enterocolitica and E coli resulted in a significant concentration- dependent increase of SCF mRNA expression. Similar results were obtained in coculture experiments with N meningitidis. As shown in experiments with E coli, the accumulation of SCF transcripts was also time-dependent. In contrast, coculture of HUVEC with the intracellular gram-negative strain C trachomatis had no effect on SCF mRNA expression. To elucidate the role of the gram-negative bacterial cell wall components, we stimulated HUVEC with bacterial lipopolysaccharide (LPS). LPS induced a maximal SCF mRNA accumulation within 2 hours followed by decrease of SCF-specific transcripts to the basal level after 24 hours. In addition, we exposed HUVEC to the classical inflammatory cytokine interleukin-1 alpha (IL-1 alpha). Kinetic experiments showed a similar pattern of regulation with an increase of SCF mRNA within 2 hours, persisting up to 12 hours, and a decrease to basal transcription after 24 hours. From these data, we conclude that SCF expression is regulated by inflammatory stimuli, such as IL-1 alpha and bacterial pathogens, suggesting an important role of SCF during inflammation.
Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting
Authors: Cameron S , Taylor DS , TePas EC , Speck NA , Mathey-Prevot B
Published: October 2019

Interleukin-3 (IL-3) is involved in proliferation and differentiation of hematopoietic progenitor cells. Its expression is subject to precise, tissue-specific regulation, and has been studied extensively in the gibbon T-cell line MLA 144 by a combination of functional assays and DNA binding experiments. To extend these studies, the gibbon IL-3 promoter was cloned and in vivo footprinting of the gibbon and human IL- 3 proximal promoters was performed. The gibbon IL-3 promoter was found to be highly homologous to its human counterpart and both promoters yielded identical in vivo footprints after induction of IL-3 synthesis. In particular, we observed specific protection of three guanines over a core sequence TGTGGTTT (IF-1IL3) that had not been recognized in previous studies. IF-1IL3 is not found in other cytokine promoters, but it is conserved in the IL-3 promoter of several species and is similar to a recurring motif in viral and T-cell-specific cellular enhancers. IF-1IL3 binds a specific complex in MLA 144 and Jurkat nuclear extracts in vitro, which shares the same specificity as the complex bound by the polyoma virus and T-cell receptor delta enhancers. Mutation of the three guanines in IF-1IL3 core sequence disrupts binding in vitro and abrogates the ability of the IL-3 promoter to mediate inducible expression in T cells. Although IF-1IL3 is necessary for IL-3 expression, it is not sufficient: a truncated IL-3 promoter with an intact IF-1IL3 site but no other activator sites is transcriptionally silent. These studies describe a new regulatory element within the IL-3 promoter that is essential for expression and conserved between species.
Circulating tumor gangliosides enhance platelet activation
Authors: Valentino LA , Ladisch S
Published: October 2019

Gangliosides enhance tumor formation in experimental animals, and high circulating concentrations of gangliosides shed by tumor cells are associated with rapid progression of human neuroblastoma. We studied these shed molecules for effects on platelet function, because platelet activation may play a role in the metastatic process. Preincubation of normal platelets in patient (tumor ganglioside-containing) serum resulted in their aggregation upon exposure to a subthreshold concentration (1 microgram/mL) of collagen (up to 34% v < 10% in normal serum) and ATP release (up to 1.1 nmol/2.5 x 10(7) platelets v < 0.2 in normal serum). Because circulating shed tumor gangliosides are lipoprotein-associated, we next assessed the effects of the serum lipoprotein fraction on platelet ATP release. The patient serum lipoprotein fraction (d > 1.210) enhanced ATP release (up to 3.1 nmol ATP), whereas the same fraction of normal serum, and both patient and control lipoprotein-depleted serum fractions (d > 1.210), were inactive (< 0.2 nmol ATP released). Finally, as little as 0.5 mumol/L patient serum gangliosides (purified from the lipoprotein fraction) caused significantly greater ATP release than did normal serum gangliosides (P < .01) and caused maximal release at 50 mumol/L (up to 3.0 nmol ATP released v < or = 0.3 nmol released by platelets exposed to normal serum gangliosides). Purified total human neuroblastoma tumor gangliosides, detected in the patient serum and isolated from LA-N5 cells, were highly active; preincubation of platelets with only 5 mumol/L of these gangliosides resulted in release of 2.5 +/- 0.1 nmol ATP. Thus, neuroblastoma patient serum, the lipoprotein fraction, and, specifically, the serum gangliosides enhance platelet activation. This activity appears to reside particularly in the tumor cell gangliosides, which are shed in vivo.
Two types of genomic rearrangements create alternative E2A-HLF fusion proteins in t(17;19)-ALL
Authors: Hunger SP , Devaraj PE , Foroni L , Secker-Walker LM , Cleary ML
Published: October 2019

The t(17;19)(q21-q22;p13.3) in acute lymphoblastic leukemia results in creation of an E2A-HLF fusion protein with structural and functional properties of a chimeric transcription factor. Two types of genomic rearrangements underlie fusion of E2A with HLF. In type I rearrangements, an insertion that codes for a portion of the chimera not found in either wild type protein occurs between E2A exon 13- and HLF exon 4-encoded sequences. This insertion is derived from a cryptic exon created at the junction between chromosomes 17 and 19, and includes intronic portions of both E2A and HLF with intervening nontemplated N nucleotides. Type II rearrangements arise from more 5′ breakpoints in E2A and result in fusion cDNAs with E2A exon 12 spliced directly to HLF exon 4. Analysis of the genomic structure of HLF shows that these different modes of protein fusion result from selective constraints to maintain the proper HLF reading frame, because a direct E2A exon 13 to HLF exon 4 splice would lead to translation of a truncated E2A protein lacking any contribution from HLF. These features underscore the requirement for DNA binding and/or dimerization conferred by the bZIP portion of the E2A-HLF chimera in t(17;19)-ALL.
Mutation His322Asn in human acetylcholinesterase does not alter electrophoretic and catalytic properties of the erythrocyte enzyme
Authors: Masson P , Froment MT , Sorenson RC , Bartels CF , Lockridge O
Published: October 2019

The YT blood group antigen is located on human red blood cell (RBC) acetylcholinesterase. Wild-type acetylcholinesterase, YT1, has histidine at codon 322, whereas the genetic variant of acetylcholinesterase, YT2, has asparagine. This mutation is located within exon 2 of the ACHE gene, an exon that is present in all alternatively spliced forms of acetylcholinesterase. Therefore, acetylcholinesterase in brain and muscle has the same mutation as RBC acetylcholinesterase. We compared the electrophoretic and kinetic properties of RBC acetylcholinesterases having His 322 or Asn 322. We found no differences in the isoelectric point, mobility on non- denaturing gel electrophoresis, affinity for acetylthiocholine, activity per milligram of RBC ghost protein, substrate inhibition constants, binding to the peripheral site ligand (propidium), and binding to active site ligands (tetrahydroaminoacridine and edrophonium). Thus, although the point mutation elicits antibody production in nonmatching blood transfusion recipients, it has no effect on the enzymatic properties of acetylcholinesterase.
Clonogenicity of circulating neuroblastoma cells: implications regarding peripheral blood stem cell transplantation
Authors: Moss TJ , Cairo M , Santana VM , Weinthal J , Hurvitz C , Bostrom B
Published: October 2019

Peripheral blood stem cells (PBSCs) are being used as an alternative to autologous marrow rescue for hematopoietic reconstitution after high- dose chemotherapy in patients with neuroblastoma and other solid malignancies. Use of PBSCs is preferred by some because of the belief that there is less risk of tumor contamination. Because tumor stem cell contamination is thought to be one contributing cause of relapse after myeloablative therapy and autologous reconstitution, we examined the potential risk of reinfusing circulating neuroblastoma cells by in vitro evaluation of their clonogenicity. Immunocytologic and tumor cell clonogenic analyses were performed on 74 blood samples obtained from 56 children with advanced-stage neuroblastoma. Concurrently drawn bone marrow specimens were evaluated in 30 instances. Circulating neoplastic cells were detected in 19 of 74 (26%) for all specimens and by immunologic techniques (26%). Using a clonogenic assay, 13 grew identifiable tumor colonies. Comparing results with the two techniques showed tumor colony growth in 10 of the 19 positive specimens by immunocytology. However, 3 of 53 samples (6%) that were negative by immunocytology were positive by the clonogenic assay. Of the 11 positive blood samples, 9 concurrent marrows contained neuroblastoma cells; of the 19 negative blood specimens, 3 concurrent marrows had metastatic disease. We conclude that circulating neuroblastoma cells are present in peripheral blood and have clonogenic properties in vitro. This supports the view that tumor cell contamination may well be one cause of relapse after autologous reconstitution. Consequently, PBSC collections should also undergo meticulous monitoring for tumor contamination before autologous reinfusion.
A Clinical Evaluation of the International Lymphoma Study Group Classification of Non-Hodgkin's Lymphoma
Authors:   The Non-Hodgkin's Lymphoma Classification Project
Published: October 2019

The recognition of several new types of non-Hodgkin's lymphoma (NHL) in recent years has led to proposals for changing lymphoma classifications, including a new proposal put forth by the International Lymphoma Study Group (ILSG). However, the clinical significance of the new entities and the practical utility of this new proposal have not been studied. Therefore, we performed a clinical evaluation of the ILSG classification. A cohort of 1,403 cases of NHL was organized at nine study sites around the world and consisted of consecutive patients seen between 1988 and 1990 who were previously untreated. A detailed protocol for histologic and clinical analysis was followed at each site, and immunologic characterization as to T- or B-cell phenotype was required. Five expert hematopathologists visited the sites and each classified each case using the ILSG classification. A consensus diagnosis was also reached in each case, and each expert rereviewed a 20% random sample of the cases. Clinical correlations and survival analyses were then performed. A diagnosis of NHL was confirmed in 1,378 (98.2%) of the cases. The most common lymphoma types were diffuse large B-cell lymphoma (31%) and follicular lymphoma (22%), whereas the new entities comprised 21% of the cases. Diagnostic accuracy was at least 85% for most of the major lymphoma types, and reproducibility of the diagnosis was 85%. Immunophenotyping improved the diagnostic accuracy by 10% to 45% for a number of the major types. The clinical features of the new entities were distinctive. Both the histologic types and the patient characteristics as defined by the International Prognostic Index predicted for patient survival. In conclusion we found that the ILSG classification can be readily applied and identifies clinically distinctive types of NHL. However, for clinical application, prognostic factors as defined by the International Prognostic Index must be combined with the histologic diagnosis for appropriate clinical decisions.
Cytokine Production Regulating Th1 and Th2 Cytokines in Hemophagocytic Lymphohistiocytosis
Authors: Osugi Yuko , Hara Junichi , Tagawa Shinichi , Takai Kenji , Hosoi Gaku , Matsuda Yoshiko , Ohta Hideaki , Fujisaki Hiroyuki , Kobayashi Michiko , Sakata Naoki , Kawa-Ha Keisei , Okada Shintaro , Tawa Akio
Published: October 2019

Hemophagocytic lymphohistiocytosis (HLH) is caused by the hyperactivation of T cells and macrophages. The clinical characteristics associated with this disease result from overproduction of Th1 cytokines including interferon-γ (IFN-γ), interleukin-2 (IL-2), and tumor necrosis factor-α (TNF-α). In this study, we analyzed the production of IL-12 and IL-4, which determine Th1 and Th2 response, respectively, and IL-10, which antagonizes Th1 cytokines, in 11 patients with HLH. IL-12 was detected in plasma in all patients (mean peak value, 30.0 ± 5.0 pg/mL), while IFN-γ was massively produced in nine patients (mean peak value, 79.2 ± 112.0 U/mL). IL-4 was not detected in any of the patients. Plasma IL10 levels were elevated in all patients (mean peak value, 2,698.0 ± 3,535.0 pg/mL). There was a positive correlation between the levels of IFN-γ and IL-10 (P < .01). The plasma concentrations of these cytokines were initially high, before decreasing after the acute phase. However, the decrease in IL-10 levels was slower than that of IFN-γ. Although the concentration of IL-12 was high at the acute phase, in some patients, a peak in the level was delayed until the chronic phase. Thus, in HLH, production of cytokines that promote development of Th1 cells appears to be predominant over that for Th2 cell development. Overproduction of IL-10 was also observed indicating that a mechanism suppressing hyperactivation of Th1 cells and monocytes/macrophages functions in patients with this disease.
Retinoid Induced Apoptosis in Leukemia Cells Through a Retinoic Acid Nuclear Receptor-Independent Pathway
Authors: Hsu C. Alex , Rishi Arun K. , Su-Li Xiao , Gerald Tonya M. , Dawson Marcia I. , Schiffer Charles , Reichert Uwe , Shroot Braham , Poirer Guy C. , Fontana Joseph A.
Published: October 2019

Trans retinoic acid (RA) has proven to be a potent therapeutic agent in the treatment of acute promyelocytic leukemia. Unfortunately, other subtypes of acute myelogenous leukemia are resistant to the antiproliferative and differentiating effects of RA. In this report, we describe a novel retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (AHPN; CD437) that not only totally inhibits the proliferation of RA-resistant leukemic cell lines HL-60R and K562 but also induces apoptosis in these cells. Exposure of HL-60R to CD437 results in the rapid (within 30 minutes) increase of the cyclin-dependent kinase inhibitor p21waf1/cip1 as well as GADD45 mRNA. Manifestations of CD437-mediated programmed cell death are noted within 2 hours, as indicated by both the cleavage and activation of the CPP32 protease and cleavage of poly (ADP-ribose) polymerase. This is followed by cleavage of bcl-2 and internucleosomal DNA degradation. HL-60R cells do not express the retinoid nuclear receptor RARβ and RARγ and express a truncated RARα. Thus, CD437 induction of p21waf1/cip1 and GADD45 mRNAs and apoptosis occurs through a unique mechanism not involving the retinoid nuclear receptors. CD437 represents a unique retinoid with therapeutic potential in the treatment of myeloid leukemia.
Spectrin St Claude, a Splicing Mutation of the Human α-Spectrin Gene Associated With Severe Poikilocytic Anemia
Authors: Fournier Catherine M. , Nicolas Gaël , Gallagher Patrick G. , Dhermy Didier , Grandchamp Bernard , Lecomte Marie-Christine
Published: October 2019

An α-spectrin variant with increased susceptibility to tryptic digestion, αII/47, was previously observed in a child with severe, recessively inherited, poikilocytic anemia. The molecular basis of this variant, spectrin St Claude, has now been identified as a splicing mutation of the α-spectrin gene due to a T → G mutation in the 3′ acceptor splice site of exon 20. This polypyrimidine tract mutation creates a new acceptor splice site, AT → AG, and leads to the production of two novel mRNAs. One mRNA contains a 12 intronic nucleotide insertion upstream of exon 20. This insertion introduces a termination codon into the reading frame and is predicted to encode a truncated protein (108 kD) that lacks the nucleation site and thus cannot be assembled in the membrane. In the other mRNA, there is in-frame skipping of exon 20, predicting a truncated (277 kD) α-spectrin chain. The homozygous propositus has only truncated 277 kD α-spectrin chains in his erythrocyte membranes. His heterozygous parents are clinically and biochemically normal. This allele was identified in 3% of asymptomatic individuals from Benin, Africa.