Red Cell Damage Research Articles

Red Cell Damage During Extracorporeal Life Support.

Sublethal damage to red blood cells (RBCs) during extracorporeal life support (ECLS) may lead to RBC loss. Using flow cytometry, phosphatidylserine-positive (PhS+) RBCs and RBC extracellular vesicles were quantified as measures of sublethal RBC injury in 41 pediatric ECLS runs, stored RBC units, and normal adult subjects. We estimated the clearance half-life of PhS+ RBCs and compared the rates of RBC loss during pediatric ECLS due to phlebotomy, intravascular hemolysis, and extravascular clearance of PhS+ RBCs. Extracorporeal life support patients had 0.9% PhS+ RBCs, sixfold higher than normal subjects (p < 0.0001). Phosphatidylserine-positive RBCs were increased in stored RBC units (twofold in whole blood derived units, p = 0.0013; 12-fold in apheresis RBC units, p < 0.0001). Phosphatidylserine-positive RBCs were cleared with an average half-life of 15 hours. During ECLS, PhS+ RBC clearance accounted for 7% of RBC loss (1-60%), phlebotomy 12%, and intravascular hemolysis 12%. Increasing PhS+ RBCs occurred in 40% of patients that died on ECLS. Red blood cell extracellular vesicles, another marker of red cell injury/activation, were elevated fivefold during ECLS. Phosphatidylserine exposure on RBCs is increased during ECLS, marking these cells for extravascular clearance with a half-life of ~15 hours and accounting for ~7% of RBC loss.

Metabolic Influences Modulating Erythrocyte Deformability and Eryptosis.

Many factors in the surrounding environment have been reported to influence erythrocyte deformability. It is likely that some influences represent reversible changes in erythrocyte rigidity that may be involved in physiological regulation, while others represent the early stages of eryptosis, i.e., the red cell self-programmed death. For example, erythrocyte rigidification during exercise is probably a reversible physiological mechanism, while the alterations of red blood cells (RBCs) observed in pathological conditions (inflammation, type 2 diabetes, and sickle-cell disease) are more likely to lead to eryptosis. The splenic clearance of rigid erythrocytes is the major regulator of RBC deformability. The physicochemical characteristics of the surrounding environment (thermal injury, pH, osmolality, oxidative stress, and plasma protein profile) also play a major role. However, there are many other factors that influence RBC deformability and eryptosis. In this comprehensive review, we discuss the various elements and circulating molecules that might influence RBCs and modify their deformability: purinergic signaling, gasotransmitters such as nitric oxide (NO), divalent cations (magnesium, zinc, and Fe2+), lactate, ketone bodies, blood lipids, and several circulating hormones. Meal composition (caloric and carbohydrate intake) also modifies RBC deformability. Therefore, RBC deformability appears to be under the influence of many factors. This suggests that several homeostatic regulatory loops adapt the red cell rigidity to the physiological conditions in order to cope with the need for oxygen or fuel delivery to tissues. Furthermore, many conditions appear to irreversibly damage red cells, resulting in their destruction and removal from the blood. These two categories of modifications to erythrocyte deformability should thus be differentiated.

P0138 / #1480: COMPARISON OF HEMOLYSIS MARKERS AFTER RBC INFUSION IN SYRINGE INFUSION PUMPS

Aims & Objectives: Syringe infusion pumps (SIP) are used to blood transfusion in some healthcare settings aiming to achieve more accurate infusions in low infusion rates as required by critically ill newborns and infants. Despite the benefits, studies have shown that SIPs can cause red cell damage, increasing the rate of hemolysis of the RBC. The objective of this study was to evaluate hemolysis markers of RBC transfused in three different brands of SIP. Methods: Lab-based experimental study with 16 aliquots of A positive RBC units from different donors. Hemolysis markers (free hemoglobin (g/dL), hemolysis ratio (%) and potassium (mmol/L)) were measured before (control) and after (SIP) RBC infusion. Three different SIP brands (SIPA, SIPB and SIPC) were compared and biomarkers were analyzed in triplicates. Results: Free hemoglobin levels (p=0.114) and hemolysis ratio (p=0.076) increased in all infusions, but without statistical significance; difference according to brands. Potassium levels increased significantly after infusion, and the SIPA caused a threefold increase when compared to SIPB and SIPC (p=0.003). Conclusions: The studied SIPs used to RBC transfusion caused significant alterations on RBC potassium levels and the performance of the devices were different between brands

Trace element status in a rare case of hemoglobin D Punjab trait

Hemoglobin D Punjab is a hemoglobin variant that occurs mainly in northwest India with prevalence rate of 1%. Homozygous form is very rare and clinically more serious than heterozygous HbD. Here we present a case of HbD trait with mild anemia and slightly altered red cell indices in a 31year old female from South India. Decreased plasma trace elements (iron, copper, zinc) and altered metalloproteinsare the novel findings, emphasizing the need for periodic assessment and prompt supplementation of micronutrients to prevent red cell damage and related complications in hemoglobinopathies.&#x0D;

Impact of different syringe pumps on red cells during paediatric simulated transfusion.

Critically ill patients frequently need blood transfusions. For safety, blood must be delivered via syringe infusion pumps, yet this can cause red cell damage and increase the rate of haemolysis. To evaluate biochemical and haemolytic markers of red blood cells transfused in three different types of syringe infusion pumps at two different infusion rates (10 and 100 mL/h). A lab-based study using aliquots of 16 red blood cell bags was undertaken. Haemolysis markers (total haemoglobin [g/dL], haematocrit [%], free haemoglobin [g/dL], potassium [mmol/L], lactate dehydrogenase [U/L], osmolality [mOsm/kg], pH, degree of haemolysis [%]) were measured before and after red blood cell infusion and exposure. Three different syringe infusion pumps brands (A, B, and C) were compared at two different infusion rates (10 and 100 mL/h). Total haemoglobin fell significantly in all red blood cell units during manipulation (pre-infusion: 26.44 ± 5.74; post-exposure: 22.62 ± 4.00; P=.026). The degree of haemolysis significantly increased by 40% after manipulation of the red blood cells. Syringe infusion pump A caused a 3-fold increase in potassium levels (3.78 ± 6.10) when compared with B (-0.14 ± 1.46) and C (1.63 ± 1.98) (P=.015). This pump also produced the worst changes, with an increase in free haemoglobin (0.05 ± 0.05; P=.038) and more haemolysis (0.08 ± 0.07; P=.033). There were significant differences and an increase in the degree of haemolysis (P=.004) at the infusion rate of 100 mL/h. Syringe infusion pumps may cause significant red blood cell damage during infusion, with increases in free haemoglobin, potassium, and the degree of haemolysis. Some pump types, with a cassette mechanism, caused more damage. In many intensive care units, bedside nurses are able to consider infusion pump choice, and understanding the impact of different pump types on red blood cells during a transfusion provides the nurses with more information to enhance decision-making and improve the quality of the transfusion.

Shear stress evaluation on blood cells using computational fluid dynamics.

Thrombus formation and hemolysis are important factors in developing blood pumps and mechanical heart valve prostheses. These phenomena are induced by flow properties. High shear stress induces platelet and red cell damage. Computational fluid dynamics (CFD) analysis calculates shear stress of fluid and particle pathlines of blood cells. We studied blood cell damage in a blood pump by using CFD analysis and proposed a method for estimating blood damage. We analyzed a pulsatile blood pump that was developed as a totally implantable left ventricular assist system at Tokai University. Shear stress on blood cells throughout pulsatile blood pumps were analyzed using CFD software. Based on the assumption that the effect of shear stress on platelets is accumulated along the trace, we proposed a method for estimating blood damage using the damage parameter D. Platelet damage parameter is calculated regardless of the division time 𝛥t which is dependent on the calculation time step. The results of the simulations are in good agreement with Giersiepen's equation obtained from the experiments. The history of shear stress on a particle was calculated using CFD analysis. The new damage parameter D yields a value close to that of Giersiepen's equation with small errors.

A chimeric recombinant protein with peroxidase and superoxide dismutase activities: Physico-chemical characterization and applicability to neutralize oxidative stress caused by ionizing radiation

A chimeric recombinant protein PSH was composed of two antioxidant enzymes, human peroxiredoxin 6 (Prx6) and superoxide dismutase (MnSOD) from Escherichia coli. Analysis of physico-chemical properties of PSH protein confirmed its high antioxidant activity. PSH was shown to protect animals from lethal and sub-lethal doses of ionizing radiation. PSH was most effective after intravenous administration short time before ionizing irradiation. Dose reduction factor for PSH comprised 1.33. PSH was effectively alleviating the degree of radiation-induced leucopenia and thrombocytopenia in animals. Beside that, PSH administration decreased DNA damage in red bone marrow cells. The chimeric recombinant PSH can be considered as an effective radioprotector for minimizing the risks of damage of animal’s body by ionizing radiation, and it could be a promising therapeutic molecule for prevention/suppression of pathological conditions accompanied or caused by oxidative stress.

Arsenic-related oxidative stress in experimentally-dosed wild great tit nestlings

Arsenic (As) is broadly distributed due to natural and anthropogenic sources, and it may cause adverse effects in birds. However, research on other elements (Pb, Hg and Cd) has been prioritized, resulting in scarce data on As exposure and related effects in wild birds. One of the mechanisms responsible for As toxicity is oxidative stress. Therefore, the aim of this study was to investigate if environmentally relevant As levels affected oxidative stress biomarkers in great tits (Parus major). This is the first field experiment studying the effects of As on oxidative stress in wild passerines. Wild great tit nestlings were orally dosed with sodium arsenite (Control: water, Low dose: 0.2 μg g−1 d−1 and High dose: 1 μg g−1 d−1; from day 3 to day 13 post-hatching). We intended to reach As concentrations similar to those at which passerines are exposed to at actual polluted areas. We compared the responses to the experimental manipulations (High, Low and Control groups) with those in an As/metal-exposed population breeding close to a Cu–Ni smelter in Finland (Smelter group). A set of antioxidants (tGSH, GSH:GSSG ratio, CAT, SOD, GST and GPx), and oxidative damage biomarkers (lipid peroxidation, protein carbonylation, 8-hydroxy-2′-deoxyguanosine formation in DNA, and telomere length) were explored in blood. Arsenic administration had no significant effect on most of the biomarkers measured: only the CAT activity was lower in the High As group and the GPx activity was enhanced in the Smelter group compared to the Control. Our results suggest that the dose and duration of the As exposure was not enough to induce oxidative damage in red cells of great tit nestlings. In spite of this, nestlings dosed with 1 μg g−1 d−1 of sodium arsenite showed non-significantly higher oxidative stress biomarkers than controls, suggesting that we were close to an effect level for the redox-defense system. Oxidative effects at equivalent As levels combined with other stressors cannot be dismissed.

Modeling and prediction of flow-induced hemolysis: a review.

Despite decades of research related to hemolysis, the accuracy of prediction algorithms for complex flows leaves much to be desired. Fundamental questions remain about how different types of fluid stresses translate to red cell membrane failure. While cellular- and molecular-level simulations hold promise, spatial resolution to such small scales is computationally intensive. This review summarizes approaches to continuum-level modeling of hemolysis, a method that is likely to be useful well into the future for design of typical cardiovascular devices. Weaknesses are revealed for the Eulerian method of hemolysis prediction and for the linearized damage function. Wide variations in scaling of red cell membrane tension are demonstrated with different types of fluid stresses when the scalar fluid stress is the same, as well as when the energy dissipation rate is the same. New experimental data are needed for red cell damage in simple flows with controlled levels of different types of stresses, including laminar shear, laminar extension (normal), turbulent shear, and turbulent extension. Such data can be curve-fit to create more universal continuum-level models and can serve to validate numerical simulations.

Clinical Significance of Detecting the Erythrocyte Life Expectancy in Patients with Malignant Lymphoma Complicated with Anemia

To investigate the clinical value of erythrocyte life span in the diagnosis and treatment of lymphoma patients with anemia. The clinical data of 45 patients with lymphoma diagnosed by Department of Hematology in Beijing Union Hospital from January 2017 to June 2017 were collected. The life of erythrocytes was detected by alveolar gas CO test. The incidence of anemia in patients with malignant lymphoma was 46.7%, 40.0% of patients displayed the shorting of blood cell life and the hemolytic status existed in 15.56% of patients. The incidence of anemia was 47.1% in patients with DLBCL. Patients with B symptoms, bone marrow involvement, splenomegaly or LDH increase often accompanied with anemia, but only the splenomegaly and LDH level were significantly negatively correlated with red blood cell life; there was a significant difference in the effects of CHOP ± R regimen and other chemotherapy regimens (R+MINE/FC/SMILE/GDP) on patients with anemia and erythrocyte life span. The incidence of anemia in patients with lymphoma is high, red cell damage or shortened life may be involved in the mechanism of anemia, the use of a strong chemotherapy regimen can significantly shorten the red blood cell life of patients and aggravates anemia.

Alzheimer's Disease: Erythrocyte 2,3-diphosphoglycerate Content and Circulating Erythropoietin.

Alzheimer's Disease (AD) features the accumulation of β-amyloid in erythrocytes. The subsequent red cell damage may well affect their oxygen-carrying capabilities. 2,3- diphosphoglycerate (2,3-DPG) binds to the hemoglobin thereby promoting oxygen release. It is theorized that 2,3-DPG is reduced in AD and that the resulting hypoxia triggers erythropoietin (EPO) release. To explore this theory, we analyzed red cell 2,3-DPG content and EPO in AD, mild cognitive impairment, and the control group, subjective cognitive impairment. We studied (i) 2,3-DPG in red cells, and (ii) circulating EPO in AD, and both markers were unaffected by dementia. Disturbances of these oxygen-regulatory pathways do not appear to participate in brain hypoxia in AD.

Influence of mechanical resistance of red cells on expressiveness of hemolysis after cardiopulmonary bypass

The study enrolled patients with ischemic heart disease with moderate (37 patients) and apparent hemolysis (18 patients) after coronary bypass surgery under cardiopulmonary bypass. Before and after the operation the concentration of free hemoglobin in blood plasma, mechanical resistance of red cells, content of TBA-active products and the general antioxidant activity in red cells were studied, while taking into account the artificial blood circulation activity and perfusion speed as well. It was established that among patients with apparent post-perfusion hemolysis the level of TBA-active products in red cells increased after surgery, while their general antioxidant activity tended to decrease. Development of moderate hemolysis is not accompanied by changes in the oxidant-antioxidant balance of red cells. There were no reliable differences in the duration of cardiopulmonary bypass, perfusion volume velocity, as well as mechanical resistance of red cells among the groups of patients at both levels of the study. Thus, individual-conditioned apparent post-perfusion hemolysis is based on a free-radical mechanism of red cell damage, and not on a mechanical trauma of blood cells when under cardiopulmonary bypass.

First Cases of Hb Agrinio Described in Patients from the Republic of Macedonia

Previous molecular analyses of α-thalassemia (α-thal) in the Republic of Macedonia have identified the following genetic defects: –α3.7 (rightward), –(α)20.5 and – –MED I deletions and Hb Icaria [α142, Term→Lys (α2), HBA2: c.427T>A] and polyadenylation signal (polyA) [AATAAA>AATGAA (α2), HBA2: c.*92A>G] point mutations. Here, we report two unrelated patients from the Romani population in the Republic of Macedonia, homozygotes for the α2-globin gene variant Hb Agrinio [α29(B10)Leu→Pro; HBA2: c.89T>C]. To date, Hb Agrinio has been described only in individuals of Greek, Cypriot and Spanish origin. Both of our patients had early presentation of the disease (3.5 years and 2 months, respectively) with frequent blood transfusions from early infancy. They have a severe intermediate phenotype of thalassemia (Hb H disease) with hemoglobin (Hb) levels of 7.8 and 7.7 g/dL, respectively. Although the HBA2: c.89T>C mutation results in an α+ allele, the severe phenotype of the homozygotes is due to the production of hyperunstable α chains that undergo post translational precipitation. This leads to a greater degree of red cell damage and hemolytic anemia. The detection of Hb Agrinio in two unrelated families of Romani ethnic origin, may suggest it is a founder mutation in this population living in the Republic of Macedonia. Considering the severity of the clinical presentation of the homozygotes or compound heterozygotes for this rare Hb variant, a targeted molecular screening for Hb Agrinio mutation carriers should be considered in all patients of Romani ethnic origin with manifested microcytosis.

Metabolomic Analysis and Antioxidant Effect of Amla (Emblica officinalis) Extract in Preventing Oxidative Stress-Induced Red Cell Damage and Plasma Protein Alterations: An In Vitro Study.

Amla (Emblica officinalis) has antidiabetic, hypolipidemic, anti-inflammatory, and antioxidant properties, but its effect on free radical induced red cell damage and membrane and plasma protein alterations has not been adequately addressed. The aim of the present study was to evaluate the antioxidant property of amla against oxidative stress-induced red cell damage and plasma protein alterations. Red blood cells (RBCs) were preincubated with different concentrations of amla extract (50, 100, 150, and 200 μg/mL) and then treated with physiological (5 mM) and pathological (50 mM) concentrations of glucose for 24 h. In another in vitro study the plasma was pretreated with different concentrations of amla extract and then incubated with 2, 2'-Azo-Bis (2-methylpropionamidine) dihydrochloride (AAPH) for 2 h. After the incubation RBC-malondialdehyde (MDA), RBC-reduced glutathione (GSH), RBC indices, RBC morphometric study, plasma MDA, protein carbonylation, total protein, and albumin were estimated. The antioxidant property of amla was assessed by DPPH assay. RBC-MDA levels were significantly decreased and RBC-GSH levels were significantly increased with higher concentration of amla extract (150 and 200 μg/mL). Red cell count and its indices were improved with the increasing concentration of amla. In addition, at higher concentration, amla restored the RBC membrane integrity. The plasma in vitro study also showed that amla improved the plasma MDA, protein carbonylation, total protein, and albumin levels. Amla extract effectively protected the RBCs and plasma proteins from the reactive oxygen species induced oxidative damage. Liquid chromatography-mass spectrometry (LC-MS) analysis of the extract revealed the presence of gallic acid, quinic acid, and quercetin as the major constituents in addition to the other flavonoids.

Autotransfusion - The effects of cell salvage on the filterability of autotransfused red cells

30 patients undergoing elective aortic surgery had intraoperative autologous blood transfusion, 15 with the Solcotrans device and 15 with the Haemonetics Cell Saver system. Markers of red cell damage and alteration of red cell parameters and biochemistry were studied. Using the St. Georges Filtrometer, the filterability of preoperative red cells was compared to that of red cells that had undergone salvage

How do we approach thrombocytopenia in critically ill patients?

A low platelet count is a frequently encountered haematological abnormality in patients treated in intensive treatment units (ITUs). Although severe thrombocytopenia (platelet count <20×109 /l) can be associated with bleeding, even moderate-degree thrombocytopenia is associated with organ failure and adverse prognosis. The aetiology for thrombocytopenia in ITU is often multifactorial and correcting one aetiology may not normalise the low platelet count. The classical view for thrombocytopenia in this setting is consumption associated with thrombin-mediated platelet activation, but other concepts, including platelet adhesion to endothelial cells and leucocytes, platelet aggregation by increased von Willebrand factor release, red cell damage and histone release, and platelet destruction by the complement system, have recently been described. The management of severe thrombocytopenia is platelet transfusion in the presence of active bleeding or invasive procedure, but the risk-benefit of prophylactic platelet transfusions in this setting is uncertain. In this review, the incidence and mechanisms of thrombocytopenia in patients with ITU, its prognostic significance and the impact on organ function is discussed. A practical approach based on the authors' experience is described to guide management of a critically ill patient who develops thrombocytopenia.

Gamma-irradiation of deglycerolized red cells does not significantly affect in vitro quality.

Red cells frozen with glycerol may require gamma-irradiation after thawing and deglycerolization for transfusion to at-risk patients. Both freezing and irradiation are known to cause red cell damage. However, the effect of irradiation on the quality of deglycerolized red cells and the optimal shelf life of such a component is currently unknown. Red cells (<7 days) were pooled, split and glycerolized using an ACP-215 automated cell washer (n = 12 pairs) and frozen at -80°C. Red cells were thawed, deglycerolized and resuspended in SAG-M. One of each pair was gamma-irradiated, while the other served as a control. Products were stored at 2-6°C and sampled for in vitro testing immediately after irradiation, and at 24 and 48 h postirradiation. Irradiation of deglycerolized red cells led to a >1·5-fold increase in extracellular potassium, compared to control units at 24 and 48 h postirradiation. Other parameters, including haemolysis, were not significantly affected by irradiation postdeglycerolization. Deglycerolized, irradiated red cells had increased supernatant potassium, but remained of acceptable quality for 24 h postirradiation.

Splenectomy for hereditary spherocytosis: complete, partial or not at all?

Hereditary spherocytosis (HS) is an hematologic disorder, common in the Northern European population, which is characterized by hemolytic anemia and splenomegaly. The pathophysiological bedrock of HS consists of the retention and damage of abnormal red cells, caused by an intrinsic erythrocyte membrane defect. Hemolysis within the spleen is the main detrminant of erythrocyte destruction in patients with HS. Splenectomy removes the primary ‘graveyard’ for spherocytes and, thus, eliminates anemia and hyperbilirubinemia and lowers the high reticulocyte number to nearly normal levels. Spleen removal is an effective therapeutic option but it is not without risks. The incidence and public health impact of spleen surgery in children with HS is substantial. Unfortunately, splenectomy is sometimes recommended erroneously for young children who have a mild clinical phenotype or is performed without relying on careful assessment of the risks and benefits. This article aims to present criticism and address controversy about HS management. Unclear and still controversial issues are tackled according to the author’s personal viewpoint, because strong evidence in different aspects of HS management is lacking.

Cesium cessation? An advantage of pathogen reduction treatments

Cesium cessation? An advantage of pathogen reduction treatments

Viscoelasticity of Hemoglobin S Gels: Moduli, Kinetics, Structure and Mechanisms

Red cell rigidification and damage due to polymerization and gelation of deoxygenated sickle cell hemoglobin lie at the root of pathogenesis in sickle cell disease. Although the abnormal blood rheology and gel viscosity have been studied, viscoelasticity has been little examined. Using sensitive cone-plate dynamic rheometry at low shear, we followed development of gels kinetically over 5 decades of increasing elasticity, G′ and loss modulus, G", until final levels were reached. Viscoelasticity progresses through 3 regimes. (1) A rapid, brief increase in G′ and G″ that occurs after initial nucleation can be explained as an initial quadratic dependence on time predicted to result from linear progress of both initiating nucleation and fiber growth with time. The percolation transition from fluid to solid viscoelasticity may also contribute to early rheological change. (2) Following stage (1), G′ and G″ increase exponentially, consistent with the dominance of heterogeneous nucleation of new fibers on the existing fiber mass. Exponential rates, B=dlnG/dt, scale as the approximate 100th power of solution hemoglobin concentration. The increasing viscoelasticity depends on polymer density, known to increase exponentially, but is also affected by two patterns of interfiber cross-linking and by domain size, packing, overlap and interdigitation. (3) Viscoelasticity reaches asymptotic levels. Final levels of G′ and G″ have a low, between linear and quadratic, power dependence on gel density. At 15 mM(heme), about 3/4 the normal red cell concentration, G′.150 KPa. This three stage sequence shows more complexity for concentrations near 14 mM heme, with changes in exponential rate during stage (2): slow exponential progress is followed by reacceleration. The slow, near-plateau, regime may depend on entanglements in a rubber-like polymeric system, but with the addition of continuing polymerization that causes residual increase in the plateau modulus, Ge.