A new antisickling agent: In vitro studies of its effect on S/S erthrocytes and on hemoglobin S

Summary 2-{benzoylamino} pyridinium benzoate (BAPB) has exhibited an antisickling effect with homozygous S/S erythrocytes in vitro. This study suggests that BAPB prevents sicklin by inhibiting the gelation of hemoglobin S.Acknowledgments. This research has been partly funded by a Sickle Cell Research grant from the State of Tennessee to Meharry Medical College. We gratefully appreciate the technical assistance of Mr Rudolf Harlan and Mrs Marjorie Deveau of the School of Graduate Studies. We offer our thanks to Dr J. W. Davis, J. D., Coordinator, Sickle Cell Project, Meharry Medical College for constant encouragement.     

The chloride effect in human hemoglobin: A new kind of allosteric mechanism

In human hemoglobin hydrogen ions, chloride, 2,3-diphosphoglycerate and CO₂ cooperate to shift the oxygen equilibrium curve to the right. Bovine hemoglobin, by contrast, has an intrinsically low oxygen affinity: when stripped, it is as low as that of human hemoglobin in the presence of 0.1 M NACl+0.1 M DPG.     

Scapharca dimeric hemoglobin: A new mechanism of information transfer between globin chains

The homodimeric hemoglobin component present in the red cells of the bivalve molluscScapharca inaequivalvis, HbI, is endowed with high cooperativity in ligand binding. This behaviour is in contrast with that of vertebrate hemoglobins in which cooperativity is associated with a tetrameric assembly and the presence of two types of chain. Analysis of the aminoacid sequence and immunological data suggested that the assembly of HbI differed from that characteristic of vertebrate hemoglobins and hence that cooperativity had an unusual structural basis. Indeed the X-ray structures of the carbonmonoxy and deoxy derivatives at 2.4 resolution showed that in HbI the heme carrying E and F helices are not exposed to solvent as in the vertebrate hemoglobin tetramer, but form the subunit interface and bring the two heme groups practically in direct contact through a network of hydrogen bonds. Ligand binding brings about marked structural changes that are limited to the heme environment, whereas quaternary changes are only minor. The structural changes in the heme environment result in alterations in the network of interactions between the heme groups which lead to changes in ligand affinity. In HbI therefore cooperativity in ligand binding is achieved through direct heme-heme communication as opposed to the long range information transfer operative in the vertebrate hemoglobin tetramer.     

Pharmacological modification of hemoglobin F expression in sickle cell anemia: An update on hydroxyurea studies

The most studied pharmacological intervention in sickle cell anemia aiming at elevating HbF expression is the use of hydroxyurea. At the present time the experience has been that after 1 year of treatment with maximum tolerated doses (MTD) all patients showed increases of percent HbF, with a mean of 15% HbF, without apparent side effects besides the reversible ones observed during the process of attaining the MTD. The question of efficacy is presently being investigated by a multicenter placebo controlled double blind clinical trial that involves more than 20 sites. The goal of the study is to determine if hydroxyurea can decrease the incidence of painful crises by 50%. Results of this study are not expected before the end of 1993.     

The in vitro antiapoptotic effect of dehydroepiandrosterone sulfate in mouse thymocytes and its relation to caspase-3/caspase-6

The effects of dehydroepiandrosterone sulfate (DHEAS) on thymocyte apoptosis induced by dexamethasone (DEX) were investigated. Apoptosis was measured by using agarose gel electrophoresis of DNA, the terminal deoxynucleotidyltransferase (TdT)- mediated dUTP nick end labeling (TUNEL) assay and flow cytometry. Our results showed that preincubation with 1×10⁻⁴ M DHEAS protected thymocytes from DEX-induced apoptosis in vitro. Moreover, we found no blocking effect on the DEX-induced activation of caspase-3 and caspase-6 by the preincubation of thymocytes with DHEAS. This may be interpreted to mean that the antagonism of DHEAS to DEX-induced apoptosis is not related to the activation of these downstream caspases which play a critical role in the execution of apoptosis. Received 25 June 1999; received after revision 1 September 1999; accepted 13 September 1999     

Penetration and interaction with haemoglobin of Corynebacteria-like microorganisms into erythrocytes in vitro

Summary Following 24 h incubation of normal blood in the presence of the microorganism, the evolution of cell wall deficient forms within the erythrocytes and a process of oxidation of the haemoglobin may be observed.     

Penetration and interaction with haemoglobin of corynebacteria-like microorganisms into erythrocytes in vitro.

Following 24 h incubation of normal blood in the presence of the microorganism, the evolution of cell wall deficient forms within the erythrocytes and a process of oxidation of the haemoglobin may be observed.     

Quantitative studies on in vitro transformation of hamster chondrocytes

Summary Hamster chondrocytes could be transformed in a quantitative assay system which used X-irradiated feeder layer cells. Morphological transformation occurred on addition of, 4NQO, but not in control cultures. Differentiation was classified into 3 types (good, poor and none); normal and transformed colonies contained similar proportions of the 3 types.This work was supported by a grant from the Ministry of Education, Science and Culture, Japan.We thank Miss M. Tanaka for her technical assistance.     

Agglutination of leukemic cells and daunomycin entrapped erythrocytes with lectin in vitro and in vivo

Summary Wheat germ agglutinin (WGA) agglutinated the L₁₂₁₀ leukemic cells and daunomycin entrapped erythrocytes in vitro. Comparing with control preparations, the greatest increase in survival was obtained in vivo when the daunomycin entrapped erythrocytes and WGA were given to BDF₁ mice bearing L₁₂₁₀ cells.     

Agglutination of leukemic cells and daunomycin entrapped erythrocytes with lectin in vitro and in vivo

Wheat germ agglutinin (WGA) agglutinated the L1210 leukemic cells and daunomycin entrapped erythrocytes in vitro. Comparing with control preparations, the greatest increase in survival was obtained in vivo when the daunomycin entrapped erythrocytes and WGA were given to BDF1 mice bearing L1210 cells.     

The influence of sodium salicylate on the formation of inorganic phosphate in human and rabbit erythrocytes in vitro

Zusammenfassung Natriumsalizylat in Konzentrationen von 2–20×10^–3 M verursacht einen bemerkenswerten Anstieg an anorganischem Phosphat sowohl in Kaninchen- wie auch in Menschenerythrozyten, die in 0,9% Natriumchlorid oder Tyrode-Locke's Lösung mehrfach gewaschen und suspendiert wurden.

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Work performed during the tenure of a Welsh Hospital Board Research Scholarship.     

H<Subscript>2</Subscript>S biosynthesis and catabolism: new insights from molecular studies

Hydrogen sulfide (H₂S) has profound biological effects within living organisms and is now increasingly being considered alongside other gaseous signalling molecules, such as nitric oxide (NO) and carbon monoxide (CO). Conventional use of pharmacological and molecular approaches has spawned a rapidly growing research field that has identified H₂S as playing a functional role in cell-signalling and post-translational modifications. Recently, a number of laboratories have reported the use of siRNA methodologies and genetic mouse models to mimic the loss of function of genes involved in the biosynthesis and degradation of H₂S within tissues. Studies utilising these systems are revealing new insights into the biology of H₂S within the cardiovascular system, inflammatory disease, and in cell signalling. In light of this work, the current review will describe recent advances in H₂S research made possible by the use of molecular approaches and genetic mouse models with perturbed capacities to generate or detoxify physiological levels of H₂S gas within tissues.