Cellular and biochemical heterogeneity contributes to the phenotypic diversity of transfusion-dependent beta thalassemia
Konstantina TheocharakiAlkmini T. AnastasiadiSophia DelicouVassilis L. TzounakasIoanna BarlaStella RouvelaEvgenia KazoliaGeorgia TzafaGeorge MpekoulisTheodore GousdovasEfthymia PavlouIoannis V. KostopoulosAthanassios D. VelentzasNikolaos SimantirisAikaterini XydakiNiki VassilakiErsi VoskaridouIoanna-Katerina AggeliEfrosyni G. NomikouOurania E. TsitsilonisEfstathia G. PapageorgiouΝikolaos S. ΤhomaidisEvagelos GikasMarianna PolitouVeroniki KomninakaMarianna H. Antonelou
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Haptoglobin
Homeostasis
Erythrocyte fragility
Objective To research whether the propyl gallate can influence the hemolysis and osmotic fragility of red blood cells. Methods Different concentrations of propyl gallate ( 256 μg/ml, 128 μg/ml, 64 μg/ml, 32 μg/ml, 16 μg/ml, 8 μg/ml) were incubated with red blood cells for 12 hours. The levels of hemolysis were detected by visual observation and hemiglobincyanide; The red blood cell osmotic fragility test was used to detected the maximum and minimum osmotic fragility. The hemiglobincyanide was also used to detect the hemolytic status of maximum osmotic fragility. The statistical software SPSS 16.0 was applied to analyze the results. Results No significant hemolysis was observed by eyes. There were no conspicuous differences of the maximum and minimum osmotic fragility among the experimental groups, the control group and the reference value of health adults. When comparing the hemolysis of the the maximum osmotic fragility, there were no significant differences (P 0.05). Conclusion Propyl gallate can't influence the hemolysis and osmotic fragility of red blood cells.
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When 2-butoxyethanol (2-BE) is administered to rats, hemolysis occurs as the active metabolite butoxyacetic acid (BAA) is formed. Human red blood cells appear to be relatively resistant to the hemolytic effects of BAA in vitro, whereas rat red blood cells undergo changes in deformability, cell swelling, and hemolysis. In this study, exposure of human red blood cells to high concentrations of BAA resulted in loss of deformability and a small increase in mean cellular volume, but no significant hemolysis. These changes resembled the changes that occur in rat erythrocytes exposed to much lower concentrations of BAA. Therefore, a comparison was made between the sub-hemolytic effects of BAA at high concentrations (up to 10 mM) on human red cells with the sub-hemolytic effects of lower concentrations of BAA (up to 0.1 mM) on rat erythrocytes. Under these conditions, human and rat erythrocyte deformability decreased, while mean cellular volume (MCV) and osmotic fragility increased. Although there was a substantial shift in rat erythrocytes to lower densities, human erythrocyte density was only slightly decreased. Human and rat erythrocyte sodium also increased. Rat erythrocytes demonstrated increased spherocytosis. In a survey of blood samples from adults and children, none demonstrated an increase in hemolysis (n = 97) or MCV (n = 65) after exposure to 10 mM BAA for 4 h. In these experiments, in which hemolysis was not evident, human erythrocytes required exposure to a 100-fold greater concentration of BAA to develop changes in red cell deformability, osmotic fragility, and sodium content similar to those observed in rat erythrocytes. These concentrations are not likely to occur under normal human use of 2-BE-containing products.
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Background: This study examines that erythrocyte was fragility-susceptible in diabetes. Methods: Forty-five outpatients with type 2 diabetes mellitus (aged 46 ± 13 years) and 20 healthy individuals with no history of diabetes disorders (aged 43 ± 7 y
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Storage of human blood for six to eight weeks in ACD solution resulted in definite hemolysis which paralleled an increase in osmotic fragility determined by a new method. Mechanical injury as a result of delivering blood by airdrop markedly aggravated this hemolysis. Whole blood stored for six weeks in ACD solution with adenine added had less overt hemolysis, less osmotic fragility and better posttransfusion survival than blood from the same subject stored only in ACD solution. Comparison of changes in the osmotic fragility of stored red blood cells showed good correlation with in vivo survival measurements and suggested that this method of testing might be useful in programs to evaluate the effect of additives on the viability of banked blood.
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Objective To observe the effects of elevated temperatures(37~44.5℃)on the cell counts, hemolysis and osmotic fragility of stored red blood cells.Methods After 7~35 days of storage at 4℃, erythrocyte suspensions were heated by incubation at 37℃ and 44.5℃ for 30 min in an incubator and then examined for cell counts, hemolysis and osmotic fragility.Results Red blood cell, white blood cell and platelet counts, hemoglobin and potassium were unchanged following the heat treatment. In the osmotic fragility test, hemolysis remained within normal limits. Hemolysis of groups incubated at 37℃ and 44.5℃ began at (4.41±0.38)g/L and (4.3±0.2)g/L respectively while a complete hemolysis occurred at (3.42±0.36)g/L and (3.4±0.1)g/L , showing no significant difference compared to the room temperature control (4.5±0.18)g/L and (3.6±0.1)g/L. There was no significant distinction in the proportions of smooth spherocyte among the samples incubated at different temperatures, except the samples stored for 35 days, probably due to the long storage period.Conclusions Heating at temperatures between 37～44.5℃ for as long as 30 min does not cause hemolysis or other damage to red blood cells.
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An osmotic fragility test is a useful way to determine the extent of red blood cell (RBC) hemolysis resulting from osmotic stress. A microfluidic chip-based system for measuring the osmotic fragility of RBCs has been developed. The chip was made from a Y-shaped polydimethylsiloxane (PDMS) microchannel sealed to a glass cover plate. Fresh rabbit blood diluted 1 : 10 with an isotonic solution and pure water respectively were introduced into the long serpentine channel using two syringe pumps. Hypotonic saline solutions with three different NaCl concentrations were prepared on a chip and images of RBCs at different locations in the channel were captured. The extent of hemolysis was estimated by comparing the cell numbers in the images using an automatic image processing program. Different degrees of hemolysis (no hemolysis, partial hemolysis, and complete hemolysis) can be estimated with this platform. This device provides a promising screening platform for diseases marked by RBC abnormalities with great simplicity, high speed and minimal requirement of blood samples.
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Continuous-flow left ventricular assist devices (LVADs) subject elements of the blood to significant stress, resulting in clinically significant and subclinical hemolysis. We sought to prospectively determine whether baseline red-cell osmotic fragility of an advanced heart-failure patient influences the hemolytic response to LVAD support. Osmotic fragility assesses the degree of red-blood-cell hemolysis under varying degrees of osmotic stress. Assays were prospectively obtained on 50 consecutive patients prior to placement of continuous-flow LVADs: HeartMate II (n = 34), Jarvik 2000 (n = 5), HeartWare (n = 6). The mean age of the patients was 60.2 years and 87% were male and 47% were nonischemic. The overall median post-LVAD lactate dehydrogenase (LDH) was 583 (427-965), and there was no difference between devices. Mean hemolysis was 15.68 ± 12.96% at 0.45% NaCl (the inflection point of the osmotic fragility hemolysis curve). A scatter plot did not reveal any relationship between preoperative osmotic fragility and postoperative LDH. Linear regression confirmed no predictive relationship (p = 0.71). In conclusion, preoperative variations in osmotic fragility do not appear to account for differences in hemolysis following ventricular assist device placement. Mechanical forces generated by existing LVADs result in similar levels of biochemical hemolysis, as assessed by LDH, despite baseline differences in a patient's osmotic red-cell fragility.
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The effects of tetrachlorobiphenyl (TCB) isomers on the osmotic fragility and shape of human erythrocytes were examined. Low concentrations of 2, 3, 2', 3'-, 2, 4, 2', 4'-and 2, 5, 2', 5'-TCB protected erythrocytes from hypotonic hemolysis, but at high concentrations, these compounds greatly promoted hypotonic hemolysis. Along with the anti-hemolytic effect, the compounds caused alterations in cell shape that ranged from normal discocytes to cup-formed cells and to spherocytes. Thus, these compounds can be classified as cup-formers. The alterations in shape were brought about by conditions in which the membranes were protected from hypotonic hemolysis. The break point in the Arrhenius plot for hypotonic hemolysis was decreased by about 3°C when 2, 3, 2', 3'-TCB was added. Therefore, these compounds increase fluidity of the bilayer, which is clearly demonstrated by the ability of 2, 3, 2', 3'-TCB to lower the break point temperature, which enables it to protect erythrocytes from osmotic swelling. By contrast, 2, 6, 2', 6'-and 3, 4, 3', 4'-TCB neither protected erythrocytes from hypotonic hemolysis nor altered cell shape. The relation between these phenomena and chemical structure is discussed.
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The specific antibody to human adult hemoglobin A1 was prepared. And the immunochemical properties of haptoglobin which also binds specifically with hemoglobin were compared with Fig. 3. Complement fixation reaction between Hb Al and (a) antihemoglobin antibody or (b) human Hp. 4: 100% hemolysis, 3: 75% hemolysis, 2: 50% hemolysis, 1: 25% hemolysis, 0: non-hemolysis. those of antihemoglobin A1 antibody. Hemoglobin-haptoglobin shows neither precipitation reaction nor complement fixation reaction under conditions in which hemoglobin-antihemoglobin antibody shows these immunological reactions.
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