scholarly journals Antisickling effect of tellurite: a potent membrane-acting agent in vitro

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 305-307 ◽  
Author(s):  
T Asakura ◽  
Y Shibutani ◽  
MP Reilly ◽  
RH DeMeio
Keyword(s):  
Hemoglobin Concentration ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Cell Swelling ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Potassium Tellurite ◽  
New Type ◽  
Acting Agent

Abstract Potassium tellurite (K2TeO3) was found to be a potent antisickling agent that inhibited red cell sickling at concentrations less than 10 mumol/L. The inhibitory effect depended on the incubation time, with the effect increasing with longer incubation periods. Because tellurite causes swelling of red cells, and because the antisickling effect of tellurite correlated with the degree of red cell swelling, the antisickling effect of tellurite is assumed to be due to the decreased mean cell hemoglobin concentration. Swelling of red cells by tellurite was accelerated by the addition of reduced glutathione. Tellurite appears to be a new type of antisickling agent that interacts with the red cell membrane.

scholarly journals Antisickling effect of tellurite: a potent membrane-acting agent in vitro

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 305-307
Author(s):  
T Asakura ◽  
Y Shibutani ◽  
MP Reilly ◽  
RH DeMeio
Keyword(s):  
Hemoglobin Concentration ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Cell Swelling ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Potassium Tellurite ◽  
New Type ◽  
Acting Agent

Potassium tellurite (K2TeO3) was found to be a potent antisickling agent that inhibited red cell sickling at concentrations less than 10 mumol/L. The inhibitory effect depended on the incubation time, with the effect increasing with longer incubation periods. Because tellurite causes swelling of red cells, and because the antisickling effect of tellurite correlated with the degree of red cell swelling, the antisickling effect of tellurite is assumed to be due to the decreased mean cell hemoglobin concentration. Swelling of red cells by tellurite was accelerated by the addition of reduced glutathione. Tellurite appears to be a new type of antisickling agent that interacts with the red cell membrane.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals The relationship between in vivo generated hemoglobin skeletal protein complex and increased red cell membrane rigidity

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1427-1431 ◽  
Author(s):  
N Fortier ◽  
LM Snyder ◽  
F Garver ◽  
C Kiefer ◽  
J McKenney ◽  
...  
Keyword(s):  
Protein Complex ◽  
Sodium Dodecyl ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Cellular Dehydration ◽  
Thalassemia Minor ◽  
Membrane Rigidity

Abstract In vitro induced oxidative damage to normal human RBCs has previously been shown to result in increased membrane rigidity as a consequence of the generation of a protein complex between hemoglobin and spectrin. In order to determine if in vivo generated hemoglobin-spectrin complexes may play a role in increased membrane rigidity of certain pathologic red cells, we measured both these parameters in membranes prepared from hereditary xerocytosis (Hx), sickle cell disease (Sc), and red cells from thalassemia minor (beta thal). Membranes were prepared from density-fractionated red cells, and membrane deformability was measured using an ektacytometer. Hemoglobin-spectrin complex was determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel analysis, as well as by Western blot analysis using a monoclonal antibody against the beta- subunit of hemoglobin. For these three types of pathologic red cells, progressive cellular dehydration was associated with increased membrane rigidity and increased content of hemoglobin-spectrin complex. Moreover, the increase in membrane rigidity appeared to be directly related to the quantity of hemoglobin-spectrin complex associated with the membrane. Our findings imply that hemoglobin-spectrin complex is generated in vivo, and this in turn results in increased membrane rigidity of certain pathologic red cells. The data further suggest that oxidative crosslinking may play an important role in the pathophysiology of certain red cell disorders.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393 ◽  
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

Abstract The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

Influence of red cell membrane on diffusion of oxygen

1960 ◽  
Vol 15 (6) ◽  
pp. 1117-1122 ◽  
Author(s):  
F. Kreuzer ◽  
W. Z. Yahr
Keyword(s):  
Cell Membrane ◽  
Light Transmission ◽  
Hemoglobin Concentration ◽  
Pure Oxygen ◽  
Red Cell ◽  
Freezing And Thawing ◽  
Red Cell Membrane ◽  
Equal Concentration ◽  
Significant Difference

Experiments in vitro were conducted on human blood at 37°C. Centrifuged blood samples were divided into packed red cells and three kinds of hemoglobin solution of equal concentration with the hemolysis effected by freezing and thawing, distilled water and ether. Since the four media had the same hemoglobin concentration, the conditions for the propagation of oxygen were substantially identical, with the only difference that the oxygen had to cross the barrier of the red cell membranes in the case of the cells. All media were analyzed with the thin-layer technique. Light transmission changes resulting from the uptake of pure oxygen by the reduced media were continuously recorded. From such tracings, the one-third and one-half saturation times were evaluated and compared. No significant difference was found between any of the highly concentrated hemoglobin solutions and the packed red cell suspensions. It was concluded that the red cell membrane does not seem to represent an essential impediment to the propagation of oxygen under these circumstances. Submitted on April 20, 1960

scholarly journals Partial spectrin deficiency in hereditary pyropoikilocytosis

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 919-924 ◽  
Author(s):  
TL Coetzer ◽  
J Palek
Keyword(s):  
Degradation Products ◽  
Membrane Skeleton ◽  
Red Cells ◽  
Proteolytic Degradation ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Erythroid Precursor ◽  
Single Defect

Abstract Hereditary pyropoikilocytosis (HPP) is a severe hemolytic anemia in which an instability of the red cell membrane skeleton has been correlated with structural and functional defects of spectrin. We now report that 13 unrelated HPP subjects have approximately 30% less spectrin than normal as evidenced by a decreased spectrin/band 3 ratio. We also examine the role of spectrin degradation as an underlying cause of this partial spectrin deficiency. Our studies demonstrate that the reduced spectrin content of HPP red cells remains constant during in vivo aging of the cells in the peripheral blood, as well as during in vitro incubation. Furthermore, immunoblotting experiments using an affinity-purified antispectrin antibody indicate that there is no loss of spectrin during membrane preparation and also that neither whole HPP red cells nor ghosts nor cytosol contains any abnormal spectrin degradation products. These data suggest that spectrin is not degraded and that it is stable on the membrane of the circulating HPP red cell. In contrast, however, incubation of free spectrin with a lysate of nucleated erythroid precursor cells indicates that HPP alpha I/46 spectrin, but not HPP alpha I/74 spectrin, is more susceptible to proteolytic degradation than a control. These data imply that the decreased spectrin content of HPP is not due to a single defect but that a more complex mechanism is involved. In HPP Sp alpha I/46 subjects, an increased proteolytic degradation in bone marrow erythroid precursors of cytosolic spectrin, prior to its assembly on the membrane, could contribute toward the partial spectrin deficiency.

scholarly journals An Unexpectedly High Frequency of Sptb Gene Mutation (SPTB exon 29: c.6055T>C (p.Ser2019Pro; Spectrin Thai)) with a Single Origin in Thailand Suggesting a New Model of Red Blood Cell Trait Against Malarial Pressure

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2322-2322
Author(s):  
Supachai Ekwattanakit ◽  
Jiraporn Korchuenjit ◽  
Thidarat Suksangpleng ◽  
Suchada Riolueang ◽  
Tana Taechalertpaisarn ◽  
...  
Keyword(s):  
Research Funding ◽  
Population Level ◽  
Extensive Study ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Thai Population ◽  
First Case ◽  
The Impact

Abstract Mutations in SPTB gene coding β-spectrin can lead to congenital hemolytic anemia (HA) or red cell membranopathy. Most patients presented with mild to moderate HA with either spherocytes or ovalo-elliptocytes. Only one report in the literature described a family with non-immune hydrops fetalis (HF) due to homozygous SPTB: c.6055T>C (p.Ser2019Pro). Recently, we have identified the second patient presented with HF in Thailand with the same genotypes through genomic approach. This finding has driven extensive study to determine the impact of SPTB mutation in our cohort of unexplained HA and HF. From 107,947 live births at our center between 2005 and 2016, 83 non-immune HF were identified. Nineteen cases from 42 unexplained HF (45%) had FFPE DNA available from autopsy (Fig 1). In addition, 7 patients who were transfusion dependent anemia due to unknown cause were recruited. These were subjects of our Whole Exome Analysis (WES) using Illumina HiSeq4000 capturing by SureSelect V5 plus UTR covering 21,522 genes. Moreover, we studied WES in 120 Thai individuals without anemia as controls. Surprisingly, 6 out of 26 patients (23%) were homozygous SPTB: c.6055T>C and none in controls. We confirmed this finding by direct genomic Sanger sequencing and set up a PCR-RFLP to detect this further in other cohorts. Interestingly, we found additional 13 heterozygotes of this mutation in our thalassemia patients (1 in 361) and general population (12 in 2,576) (Fig 1) making an unexpectedly high carrier rate of this "so-called rare genetic disease" into 0.47% in Thai population. Using Hardy-Weinberg principle, homozygous SPTB: c.6055T>C would be 5.42 per million or expected 358 cases in Thailand. Since most reported SPTB mutations are sporadic or familial specific, we wondered whether this affected allele has a single or multiple origins. We generated a haplotype analysis of SPTB on chromosome 14q23 using 23 informative SNPs at the gene locus and adjacent region from WES data. Total 23 different haplotypes were generated and the SPTB c.6055T>C originated from the same ancestral chromosome as it was found only in haplotype IA (Fig 2). Therefore, we proposed to name this as spectrin Thai, instead of spectrin Providence which was named after the place of the first case reported (Gallagher PG, et al. J Clin Invest 1995), but not related to its native population. All 4 survived patients due to homozygous spectrin Thai (one received intrauterine transfusion) had severe anemia at birth and neonatal jaundice requiring therapy and became transfusion dependent afterward. All had marked hepatosplenomegaly with blood picture of pyropoikilocytosis. Three patients underwent splenectomy without improvement on anemia, this finding contradicted those in patients with red cell membranopathies. Of note, hematology from 25 heterozygotes including 12 parents showed no apparently abnormal indices and normal EMA binding assay. Using shortgun proteomics to analyze red cell membrane proteins, hemolysate and plasma proteins, the only difference was we could detect unbound free β-spectrin in plasma from controls but not from spectrin Thai traits. This might be useful for future screening of heterozygotes at population level. To explore whether red cells with spectrin Thai trait has any protection against malaria, the most important selective pressure in the past of our region we studied in vitro growth of P. falciparum and kinetics of erythroid invasion using live-cell microscopy. We found no significant differences between normal and spectrin Thai traits on these aspects. Since β-spectrin is involved on knob formation of malarial infected red cells that plays an important role on sequestration and cerebral manifestation, a further study using scanning electron microscopy comparing between malarial infected normal against spectrin Thai red cells is underway. For the first time, we showed that red cell membranopathy might be endemic in the tropics than we expected. Homozygous spectrin Thai is one of the most common causes of unexplained HF and HA in our region. The possibility that red cell membranopathy traits might be under malarial pressure throughout human history has completed the whole red cell quantitative trait genetic polymorphism in our population. This is in consistent with those of hemoglobinopathy (thalassemia) and enzymopathy (G6PD), both are highly prevalent in our region due to the same biological advantage against malaria. Disclosures Viprakasit: Agios: Consultancy, Research Funding; Protagonist Therapeutics: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; F. Hoffmann-La Roche Ltd: Consultancy, Research Funding.

scholarly journals Partial spectrin deficiency in hereditary pyropoikilocytosis

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 919-924
Author(s):  
TL Coetzer ◽  
J Palek
Keyword(s):  
Degradation Products ◽  
Membrane Skeleton ◽  
Red Cells ◽  
Proteolytic Degradation ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Erythroid Precursor ◽  
Single Defect

Hereditary pyropoikilocytosis (HPP) is a severe hemolytic anemia in which an instability of the red cell membrane skeleton has been correlated with structural and functional defects of spectrin. We now report that 13 unrelated HPP subjects have approximately 30% less spectrin than normal as evidenced by a decreased spectrin/band 3 ratio. We also examine the role of spectrin degradation as an underlying cause of this partial spectrin deficiency. Our studies demonstrate that the reduced spectrin content of HPP red cells remains constant during in vivo aging of the cells in the peripheral blood, as well as during in vitro incubation. Furthermore, immunoblotting experiments using an affinity-purified antispectrin antibody indicate that there is no loss of spectrin during membrane preparation and also that neither whole HPP red cells nor ghosts nor cytosol contains any abnormal spectrin degradation products. These data suggest that spectrin is not degraded and that it is stable on the membrane of the circulating HPP red cell. In contrast, however, incubation of free spectrin with a lysate of nucleated erythroid precursor cells indicates that HPP alpha I/46 spectrin, but not HPP alpha I/74 spectrin, is more susceptible to proteolytic degradation than a control. These data imply that the decreased spectrin content of HPP is not due to a single defect but that a more complex mechanism is involved. In HPP Sp alpha I/46 subjects, an increased proteolytic degradation in bone marrow erythroid precursors of cytosolic spectrin, prior to its assembly on the membrane, could contribute toward the partial spectrin deficiency.

Abnormal Erythrocyte Endothelial Adherence in Hereditary Stomatocytosis

Blood ◽  
1997 ◽  
Vol 89 (9) ◽  
pp. 3451-3456 ◽  
Author(s):  
Brian D. Smith ◽  
George B. Segel
Keyword(s):  
Clinical Course ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Endothelial Monolayers ◽  
Cell Membrane Protein ◽  
Asymptomatic Sibling ◽  
Normal Controls

Abstract Hereditary stomatocytosis is a red cell membrane protein disorder, which results in hemolytic anemia. Some patients with hereditary stomatocytosis experience dyspnea, chest pain, and abdominal pain, particularly after splenectomy. These symptoms may represent vaso-occlusion secondary to adherence of an abnormal erythrocyte membrane to vascular endothelium. We studied three members of a family with varying clinical expression of hereditary stomatocytosis. Adherence of red cells to endothelium was quantified by measuring the shear force required to separate individual cells from endothelial monolayers using a micropipette technique. Two patients with symptoms of in situ thromboses had a higher percentage of adherent cells compared with their asymptomatic sibling and normal controls. Correlation between this in vitro phenomenon and the clinical course suggests that flow abnormalities in the microcirculation attributable to erythrocyte endothelial adherence may play an important pathogenetic role in the illness. When the proportion of adherent red cells was reduced by a chronic transfusion program in one patient and pentoxifyllin therapy in another, the vaso-occlusive complications were eliminated.

scholarly journals Diisopropylfluorophosphate Is Not a Specific Label for the Red Cell Membrane

Blood ◽  
1969 ◽  
Vol 34 (3) ◽  
pp. 376-379 ◽  
Author(s):  
DAVID A. SEARS ◽  
ROBERT I. WEED
Keyword(s):  
Cell Membrane ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Membrane Enzyme ◽  
Specific Label

Abstract When red cells were labeled either in vitro or in vivo with DF32P, the label was attached primarily at intracellular, not membrane, sites. Thus, despite the fact that it binds to and inactivates the red cell membrane enzyme, acetylcholinesterate, DF32P is not a specific label for the red cell membrane.

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