Alteration In Osmotic Fragility Research Articles

Imidacloprid induced osmotic fragility in erythrocytes of rats: Protective role of vit.C and tea

The present study was undertaken to evaluate the effect in vitro induction of sub-acute oral dose of imidacloprid and ameliorating role of tea and Vit.C on oxidative stress and osmotic fragility of albino wistar rats. On the basis of imidacloprid having LD50 450mg/kg body weight in rats, which exert oxidative stress on mammalian system and is moderately toxic via oral route. We observed a significant alteration in osmotic fragility of rat erythrocytes upon imidacloprid treatment shown by mean erythrocyte osmotic fragility at relatively higher NaCl concentration of (0.45%) as compared to mean erythrocyte fragility(MEF) at 0.40%,0.41% 42%,0.32% and 0.30% NaCl in control, Vit. C, tea, IP+ tea, IP + Vit.C treated groups, respectively. The results suggest that imidacloprid induced toxicity exerts OF in erythrocytes of rats and pretreatment with Vit. C and tea can mitigate to some extent the toxic effects.

Alterations in osmotic fragility of the red blood cells in hypo- and hyperthyroid patients

Changes in concentration of thyroid hormones can affect Na+-K+-ATPase number and activity and phospholipid composition of the cell membranes leading to changes in the surface to volume ratio and strength of membrane. In this study, the osmotic fragility of the red blood cells from non-treated hypo- and hyperthyroid patients was compared to that of control subjects. After 3 washings with normal saline, red blood cells were placed in varying concentrations of sodium chloride (Na- Cl) (0-0.9%) and fragility was assessed with colorimetric method; to do this, after the incubation period, tubes were centrifuged and the optical density of the tubes was measured. Hemolysis percentage in tubes was calculated based on 100% hemolysis in the tubes containing no NaCl (0%). Osmotic fragility of the cells from hyperthyroid patients in 0.45% NaCl was significantly lower than control subjects (74.6%+/-30.2 vs 93.8%+/-9.1, p<0.01). The osmotic fragility of red blood cells in 0.5% concentration of sodium chloride in hyperthyroid patients was significantly lower compared to that of controls (27.8%+/-26.0 vs 63.5%+/-27.5, p<0.001). No significant difference was observed between the osmotic fragility of the hypothyroid patients compared with control subjects. Alteration in osmotic fragility is seen in patients with hyperthyroidism; however, anemia reported in hypo- or hyperthyroid patients is not due to high osmotic fragility of red blood cells and other causes need to be investigated.

Mild spherocytic hereditary elliptocytosis and altered levels of α- and γ-adducins in β-adducin-deficient mice

AbstractThe membrane skeleton, a dynamic network of proteins associated with the plasma membrane, determines the shape and mechanical properties of erythrocytes. Deficiencies or defects in membrane skeletal proteins are associated with inherited disorders of erythrocyte morphology and function. Adducin is one of the proteins localized at the spectrin-actin junction of the membrane skeleton. In this work we show that deficiency of β-adducin produces an 80% decrease of α-adducin and a fourfold up-regulation of γ-adducin in erythrocytes. β-Adducin or any other isoform generated by translation of abnormally spliced messenger RNAs could not be detected by our antibodies either in ghosts or in cytoplasm of −/− erythrocytes. Actin levels were diminished in mutant mice, suggesting alterations in the actin-spectrin junctional complexes due to the absence of adducin. Elliptocytes, ovalocytes, and occasionally spherocytes were found in the blood film of −/− mice. Hematological values showed an increase in reticulocyte counts and mean corpuscular hemoglobin concentration, decreased mean corpuscular volume and hematocrit, and normal erythrocyte counts that, associated to splenomegaly, indicate that the mice suffer from mild anemia with compensated hemolysis. These modifications are due to a loss of membrane surface and dehydration that result in an increase in the osmotic fragility of red blood cells. The marked alteration in osmotic fragility together with the predominant presence of elliptocytes is reminiscent of the human disorder called spherocytic hereditary elliptocytosis. Our results suggest that the amount of adducin remaining in the mutant animals (presumably αγ adducin) could be functional and might account for the mild phenotype.

Mild spherocytic hereditary elliptocytosis and altered levels of α- and γ-adducins in β-adducin-deficient mice

The membrane skeleton, a dynamic network of proteins associated with the plasma membrane, determines the shape and mechanical properties of erythrocytes. Deficiencies or defects in membrane skeletal proteins are associated with inherited disorders of erythrocyte morphology and function. Adducin is one of the proteins localized at the spectrin-actin junction of the membrane skeleton. In this work we show that deficiency of β-adducin produces an 80% decrease of -adducin and a fourfold up-regulation of γ-adducin in erythrocytes. β-Adducin or any other isoform generated by translation of abnormally spliced messenger RNAs could not be detected by our antibodies either in ghosts or in cytoplasm of −/− erythrocytes. Actin levels were diminished in mutant mice, suggesting alterations in the actin-spectrin junctional complexes due to the absence of adducin. Elliptocytes, ovalocytes, and occasionally spherocytes were found in the blood film of −/− mice. Hematological values showed an increase in reticulocyte counts and mean corpuscular hemoglobin concentration, decreased mean corpuscular volume and hematocrit, and normal erythrocyte counts that, associated to splenomegaly, indicate that the mice suffer from mild anemia with compensated hemolysis. These modifications are due to a loss of membrane surface and dehydration that result in an increase in the osmotic fragility of red blood cells. The marked alteration in osmotic fragility together with the predominant presence of elliptocytes is reminiscent of the human disorder called spherocytic hereditary elliptocytosis. Our results suggest that the amount of adducin remaining in the mutant animals (presumably γ adducin) could be functional and might account for the mild phenotype.

Haematological Parameters and Altered Erythrocyte Metabolism in Anaemic Dogs

In previous studies, an increase in the activity of pyruvate kinase (PK) and glucose-6-phosphate dehydrogenase (G6PDH), an increase in the concentration of 2,3-diphosphoglycerate (2,3DPG), and alterations in osmotic fragility were found in dogs with haemolytic anaemia. These changes were mainly caused by the presence of immature red cells found in regenerative anaemias. In the present study, the same parameters were evaluated in dogs with different types of anaemia. The haematological patterns of 40 anaemic dogs were analysed to define the pathogenesis and the haematological features of each case. Non-regenerative anaemias could be attributed principally to chronic diseases and to the haemolysis that accompanies the early stages of canine babesiosis. Regenerative anaemias were mainly due to haemolysis, in some cases with an immune-mediated pathogenesis. PK activity was higher in regenerative than in non-regenerative anaemias, but G6PDH activity and 2,3DPG concentration increased in both types of anaemia. This suggests that PK activity is influenced by the presence of immature red cells, but the requirements for reducing compounds and oxygen are not dependent on the type of anaemia. Abnormalities in osmotic fragility were detected in haemolytic anaemias and in those non-regenerative anaemias in which reticulocyte percentage, but not reticulocyte production index (RPI), increased. The osmotic fragility could be used as an early indicator of erythrocyte regeneration.

Relationship of membrane fluidity, chemoprotection, and the intrinsic toxicity of butylated hydroxytoluene

In isolated rat hepatocytes, many chemicals elicit toxicity which is inhibitable by antioxidants such as butylated hydroxytoluene (BHT). Although BHT protection is evident at concentrations of less than about 50 nmol/mg protein, higher concentrations exhibit intrinsic concentration-dependent toxicity, which involves mitochondrial dysfunction. We evaluated the possibility that both chemoprotection and intrinsic toxicity could be explained by a common mechanism involving alterations in the physical properties of cellular membranes. In the red blood cell (RBC) osmotic fragility assay, BHT at less than 60 nmol/mg protein protected against osmotic fragility; however, BHT at higher concentrations enhanced osmotic fragility such that total osmolysis occurred at 135 nmol/mg. The BHT-mediated alterations in osmotic fragility correlated with changes in membrane fluidity, determined by fluorescence polarization of the hydrophobic probe 1,6-diphenyl-1,3,5-hexatriene. Protection from osmolysis correlated with decreased fluidity, while enhanced RBC fragility correlated with increased fluidity. In rat hepatocyte suspensions, high BHT concentrations also permeabilized the plasma and mitochondrial membranes to enzyme leakage, and these effects were accompanied by enhanced membrane fluidity. Although other mechanisms may be operative, alterations in membrane fluidity appear to be, in part, responsible for the observed chemoprotective effects at low concentrations, and intrinsic toxicity at higher concentrations of BHT.