Using a wide range of different physical and chemical methods, it was found that the oxidative stress caused by addition of hydrogen peroxide to the incubation medium has a significant effect on the conformation of haematoporphyrin, influencing the oxygen-binding properties of haemoglobin in red blood cells. Morphofunctional characteristics of red blood cells change; in particular, we have observed the transformation of erythrocytes, their transition into echinocytes. In erythrocytes, in response to increased lipid peroxidation (LPO) antioxidant enzymes become active. The use of natural antioxidants (β-carotene and resveratrol) works towards reducting the level of oxidative processes. Resveratrol has the greatest antioxidant effect.
The aim of this study was to investigate the effect of hyperoxia, calcium ions and pH value on the composition of major phospholipids in human erythrocyte membranes and erythrocytes' oxygentransport function. To create a model of hyperoxia, we saturated the incubated mixture with oxygen by constant passing of oxygen-air mixture through the incubation medium. To assess the effect of elevated calcium ion concentrations, CaCl 2 was added to the incubation medium. An incubation medium with different pH was used to study the effect of various pH values. Lipids were extracted from erythrocytes and chromatographic separation was carried out in a thin layer of silica gel deposited on a glass plate. The thiobarbituric acid (TBA)-active products and the content of diene conjugates (DC) in erythrocytes were determined. The oxygen-binding capacity of haemoglobin was evaluated using Raman spectroscopy. The obtained results indicated that hyperoxia causes deep changes both in the composition and character of bilayer lipids of erythrocyte membranes, which affects the functional characteristics of erythrocytes, primarily the oxygen-transport properties of erythrocyte haemoglobin. It should be noted that a combination of Ca 2+ ions and change in the pH value intensify the processes associated with disruption of phospholipids' composition. The findings indicate that the lipid phase is one of the key elements in the functioning of erythrocytes in norm as well as during development of various pathological processes.
The aim of the investigation was to study differential gene expression and signaling pathways activation in radioresistant and radiosensitive cancer cell lines.Materials and Methods. K562 radioresistant cancer cell line of chronic myeloleukosis, and HCT-116p53 (+/+), HCT-116p53 (-/-), Me45 radiosensitive cancer cell lines were used in the study. Transcriptome analysis was performed using Affymetrix DNA microarray HGU133A series. Original Gene Selector program was used for bioinformation analysis. Protein interaction networks were studied by means of online STRING 9.0 system. To range the genes according to the signaling pathways the PANTHER program and QIAGEN SABiosciences database were used. Quantitative activation of signaling pathways was calculated with the help of OncoFinder bioinformation algorithm.Results. Four genes (DAAM1, IFNAR2, PALLD, STK17A) were identified to increase their expression in radioresistant cell line and to decrease it in radiosensitive cell lines under radiation exposure. Three signal pathways -Wnt, Interferon and p53 -in which these genes are involved and which are common to the examined cell lines were found using PANTHER program. A significant difference in activation of these signaling pathways was discovered in radioresistant and radiosensitive cell lines.Conclusion. Based on the analysis of molecular interaction networks and signaling pathway activation it has been revealed that the greatest differences between radioresistant and radiosensitive cell lines are observed in the activity of Interferon and p53 signaling pathways. TP53 gene expression level is not associated with the activity of p53 signaling pathway. Comparison of gene expression changes and activation of signaling pathways allows us to suggest the expression of DAAM1, IFNAR2, PALLD genes as indicators of radioresistance under radiation exposure.
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