Abstract
Eryptosis is the suicidal destruction-process of erythrocytes, much like apoptosis of nucleated cells, in the course of which the stressed red cell undergoes cell-shrinkage, vesiculation and externalization of membrane phosphatidylserine. Currently, there exist numerous methods to detect eryptosis, both morphometrically and biochemically. This study aimed to design a simple but sensitive, automated computerized approach to instantaneously detect eryptotic red cells and quantify their hallmark morphological characteristics. Red cells from 17 healthy volunteers were exposed to normal Ringer and hyperosmotic stress with sodium chloride, following which morphometric comparisons were conducted from their photomicrographs. The proposed method was found to significantly detect and differentiate normal and eryptotic red cells, based on variations in their structural markers. The receiver operating characteristic curve analysis for each of the markers showed a significant discriminatory accuracy with high sensitivity, specificity and area under the curve values. The software-based technique was then validated with RBCs in malaria. This model, quantifies eryptosis morphometrically in real-time, with minimal manual intervention, providing a new window to explore eryptosis triggered by different stressors and diseases and can find wide application in laboratories of hematology, blood banks and medical research.
Highlights
Erythrocyte is the simplest of all human cells, but endowed with the pivotal role of carrying oxygen from lungs to tissues for metabolism (Jensen, 2009) A decrease in the number of circulating red blood cells (RBCs), due to defect, injury or disease, hampers the oxygen-carrying capacity of blood, resulting in anemia
There are numerous diseases and conditions that are found to have an association with accelerated eryptosis, some of them being iron deficiency anemia (Kempe et al, 2006), malaria (Föller et al, 2009a), bacterial sepsis (Kempe et al, 2007), sickle cell anemia (Weiss et al, 2012), β-thalassemia (Basu et al, 2010), hemolytic uremic syndrome (Lang et al, 2006), glucose-6phosphate dehydrogenase deficiency (Lang et al, 2002), diabetes (Fırat et al, 2012) and renal insufficiency (Abed et al, 2014)
Normocytic RBCs appeared to be discoid with a central halo and had no membrane vesiculations (Figure 1A)
Summary
Erythrocyte is the simplest of all human cells, but endowed with the pivotal role of carrying oxygen from lungs to tissues for metabolism (Jensen, 2009) A decrease in the number of circulating red blood cells (RBCs), due to defect, injury or disease, hampers the oxygen-carrying capacity of blood, resulting in anemia. RBCs are removed from circulation by 120 days (Piomelli and Seaman, 1993). Sometimes, it can undergo a premature programmed apoptotic-like death, termed eryptosis, even though it is devoid of a nucleus and mitochondria (Bosman et al, 2005; Lang K.S. et al, 2005). In such cases, RBCs are cleared off from the bloodstream much earlier than its physiological lifespan, limiting the deleterious consequences of intravascular hemolysis (Kiefer and Snyder, 2000). There are numerous diseases and conditions that are found to have an association with accelerated eryptosis, some of them being iron deficiency anemia (Kempe et al, 2006), malaria (Föller et al, 2009a), bacterial sepsis (Kempe et al, 2007), sickle cell anemia (Weiss et al, 2012), β-thalassemia (Basu et al, 2010), hemolytic uremic syndrome (Lang et al, 2006), glucose-6phosphate dehydrogenase deficiency (Lang et al, 2002), diabetes (Fırat et al, 2012) and renal insufficiency (Abed et al, 2014)
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