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The aim of this study was to evaluate changes in RBC aggregation and deformability over 24 hr and suggest specific shear stress values for detecting RBC deformability in a mouse endotoxemia model using lipopolysaccharide (LPS). Six-week-old male BALB/c mice received LPS (20 mg/kg) intraperitoneally. Aggregation indices (AIs) and T1/2 were measured to assess RBC aggregation, and elongation indices (EIs) were used to assess RBC deformability at shear stress values of 0.3, 0.5, 1, 3, 7, 10, 15 and 20 Pascals (Pa) 0, 30 min, 1, 2, 4, 6, 9, 12, 18 and 24 hours after the LPS injection. No significant differences were detected in the AIs during the study period, however, T1/2 shortened significantly 2, 6, 12, 18, and 24 hr after the LPS injection. The EIs increased significantly 24 hr after LPS injection at 0.5 and 1 Pa shear stress, whereas it decreased significantly at 10 Pa of shear stress 24 hr after the LPS injection. Altered RBC deformability was detectable 24 hr after the LPS injection and T1/2 may be a sensitive marker for detecting changes in RBC aggregation. The EIs should be measured at 1 Pa to detect changes in RBC deformability in LPS-induced septic mice. [ABSTRACT FROM AUTHOR]

This study examines the effects of endurance training on red blood cells (RBC) in seventeen non-insulin-dependent type 2 diabetic men with a special focus on in vivo RBC aging. Venous blood was collected pre- and post-training at rest. RBC from whole blood and RBC separated according to cell age by density-gradient centrifugation were analyzed. RBC deformability was measured by ektacytometry. Immunohistochemical staining was performed to quantify the RBC-nitric oxide (NO) synthase activation (RBC-NOSSer1177) because RBC-NOS-produced NO can contribute to increased RBC deformability. The proportion of "young" RBC was significantly higher post-training. RBC deformability of all RBC (RBC of all ages) remained unaltered post-training. During RBC aging, RBC deformability decreased in both pre- and post-training. However, the training significantly increased RBC deformability in "young" and reduced their deformability in aging RBC. RBC-NOS activation remained unaltered in all RBC post-training. It tendentially increased in aging RBC pre-training, but did not change during aging post-training. The training significantly reduced RBC-NOS activation in "old" RBC. Endurance training may improve the RBC system (higher amount of "young" RBC which are more deformable). It remains speculative whether changes in older RBC (reduced RBC-NOS activation and deformability) could lead to more rapid elimination of aged RBC. [ABSTRACT FROM AUTHOR]

Significant progress has been made over the years on the topic of hemorheology, not only in terms of the development of more accurate and sophisticated techniques, but also in terms of understanding the phenomena associated with blood components, their interactions and impact upon blood properties. The rheological properties of blood are strongly dependent on the interactions and mechanical properties of red blood cells, and a variation of these properties can bring further insight into the human health state and can be an important parameter in clinical diagnosis. In this article, we provide both a reference for hemorheological research and a resource regarding the fundamental concepts in hemorheology. This review is aimed at those starting in the field of hemodynamics, where blood rheology plays a significant role, but also at those in search of the most up-to-date findings (both qualitative and quantitative) in hemorheological measurements and novel techniques used in this context, including technical advances under more extreme conditions such as in large amplitude oscillatory shear flow or under extensional flow, which impose large deformations comparable to those found in the microcirculatory system and in diseased vessels. Given the impressive rate of increase in the available knowledge on blood flow, this review is also intended to identify areas where current knowledge is still incomplete, and which have the potential for new, exciting and useful research. We also discuss the most important parameters that can lead to an alteration of blood rheology, and which as a consequence can have a significant impact on the normal physiological behavior of blood. [ABSTRACT FROM AUTHOR]

Microvascular circulation plays a vital role in regulating physiological functions, such as vascular resistance, and maintaining organ health. Pathologies such as hypertension, diabetes, or hematologic diseases affect the microcirculation posing a significant risk to human health. The retinal vasculature provides a unique window for non-invasive visualisation of the human circulation in vivo and retinal vascular image analysis has been established to predict the development of both clinical and subclinical cardiovascular, metabolic, renal and retinal disease in epidemiologic studies. Blood viscosity which was otherwise thought to play a negligible role in determining blood flow based on Poiseuille's law up to the 1970s has now been shown to play an equally if not a more important role in controlling microcirculation and quantifying blood flow. Understanding the hemodynamics/rheology of the microcirculation and its changes in diseased states remains a challenging task; this is due to the particulate nature of blood, the mechanical properties of the cells (such as deformability and aggregability) and the complex architecture of the microvasculature. In our review, we have tried to postulate a possible role of red blood cell (RBC) biomechanical properties and laid down future framework for research related to hemorrheological aspects of blood in patients with retinal vascular disorders. [ABSTRACT FROM AUTHOR]

BACKGROUND: Postmenopausal women often develop hemorheological disorders which may affect the systemic blood circulation and present a cardiovascular risk factor. OBJECTIVE:We evaluated effects of secoisolariciresinol (SECO), a phytoestrogen, on hemorheological parameters and lipid peroxidation in a model of the age-related and/or surgical menopause induced by ovariectomy in rats. METHODS: Arterial blood was sampled from sham-operated female rats, ovariectomized rats (OVX), and OVX treated with SECO (OVXSECO) (20 mg/kg/day intragastrically for two weeks). Plasma estrogen concentration and the following hemorheological parameters were measured: RBC aggregation (half-time of aggregation, T1/2; amplitude of aggregation, AMP; aggregation index, AI), RBC deformability (elongation index, EI), whole blood viscosity at the shear rate of 3-300 s-1, plasma viscosity, hematocrit, plasma fibrinogen. Lipid peroxidation was evaluated by measuring conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS) in plasma. RESULTS: Ovariectomy in rats caused a 60% decrease in plasma estrogen level and triggered the development of macro- and microhemorheological abnormalities. Blood viscosity increased by 12-31%, RBC elongation index reduced by 16-28%, and T1/2 and AI increased by 35% and 29% respectively. The increase in blood viscosity correlated predominantly with reduced RBC deformability. Plasma CD and TBARS were elevated by 47% and 104% respectively. SECO therapy for OVX rats reduced blood viscosity by 9-18% and T1/2 by 32%, and increased EI by 4-17%. SECO therapy disrupted the correlation between blood viscosity and RBC deformability. Lipid peroxidation was significantly inhibited, as shown by the reduction in CD and TBARS plasma concentrations by 89% and 70% respectively. SECO did not affect plasma viscosity, estrogen or fibrinogen levels. CONCLUSIONS: SECO treatment for OVX rats improves blood macro- and microrheological parameters, possibly through antioxidant protection of RBC. [ABSTRACT FROM AUTHOR]

A key challenge in transfusion medicine research and clinical hematology is to develop a simple and non-destructive method to measure the quality of each blood unit prior to use. RBC deformability has long been proposed as an indicator of blood quality. We measured RBC deformability using the pressure required for single cells to transit through a micrometer scale constriction to examine longitudinal changes in RBC deformability, as well as the variability in blood quality and storage capacity across donors. We used a microfluidic device to monitor deformability changes in RBCs stored in plastic tubes and in blood bags over 14 and 56 days respectively. We found consistent storage based degradation of RBC deformability with statistically significant variability in both the initial RBC deformability and storage capacity among donors. Furthermore, all samples exhibited a transient recovery phenomenon. Deformability profiling of stored RBCs using transiting pressure showed significant donor variability in initial quality and storage capacity. This measurement approach shows promise as a rapid method to individually assess the quality of stored RBC units. [ABSTRACT FROM AUTHOR]

The article presents a study conducted to the find out the effects of electrostimulation of the whole body (WB-EMS) on the deformability of human red blood cells (RBC). Topics discussed includes improvement in the strength of trained and untrained athletes by performing WB-EMS, assessment of performance of athletes by maximal strength test and increased nitric oxide production due to red blood cell-nitric oxide synthase (RBC-NOS) enzyme.

Rheohaemapheresis aims to normalize major rheological parameters and is used to treat patients with dry age-related macular degeneration (AMD). While effective, this approach is invasive and requires specially trained personnel. Therefore, the search for novel effective compounds with hemorheological properties that can be taken orally to treat AMD is justified. The use of a robust rodent model of AMD with high blood viscosity is crucial to test the efficacy of potential hemorheological drugs to treat this disease. The objective of this study was to investigate whether OXYS rats, generally used as an animal model of AMD, have hyperviscosity syndrome. The results of this study show that blood viscosity in OXYS rats at low (3-10 s-1) and high (45-300 s-1) shear rates were 14-20% and 7-10% higher than in Wistar rats, while hematocrit and plasma viscosity were not different. Red blood cells (RBCs) in OXYS rats were more prone to aggregation as shown by 39% shorter half-time than in Wistar rats. RBCs were also more rigid in OXYS than in Wistar rats as shown by 21-33% lower index of elongation at the shear stress of 1-7 Pa. These data indicate that OXYS rats have hyperviscosity syndrome as the result of abnormal RBC deformability and aggregation. We propose to use OXYS rats as an animal model for preclinical studies to test compounds with hemorheological properties aimed to treat AMD. [ABSTRACT FROM AUTHOR]

• Blood microflows in a micropillar array are studied. • Healthy and hardened red blood cell (RBCs) suspensions are examined and their flows compared. • The velocity field is resolved by micro Particle Image Velocimetry techniques. • RBC distributions are inferred from image intensity. • Impact of RBC deformability more evident on haematocrit distributions rather than velocity field. Red blood cell (RBC) deformability is an important haemorheological factor; it is impaired in many pathologies leading to microvascular complications. Several microfluidic platforms have been utilized to examine the role of deformability in RBC flows but their geometries tend to be simplified. In the present study, we extend our previous work on healthy RBC flows in micropillar arrays [1] to probe the effect of impaired RBC deformability on the velocity and haematocrit distributions in microscale RBC flows. Healthy and artificially hardened RBC suspensions at 25% haematocrit were perfused through the micropillar array at various flow rates and imaged. RBC velocities were determined by Particle Image Velocimetry (PIV) and haematocrit distributions were inferred from the image intensity distributions. The pillars divide the flow into two distinct RBC streams separated by a cell-depleted region along the centreline and in the rear/front stagnation points. RBC deformability was not found to significantly affect the velocity distributions; the shape of the velocity profiles in the interstitial space remained the same for healthy and hardened RBCs. Time-averaged and spatiotemporal intensity distributions, however, reveal differences in the dynamics and local distributions of healthy and hardened cells; hardened cells appear to enter the cell-depleted regions more frequently and their interstitial distributions are more uniform. The study highlights the importance of local RBC distributions and the impact of RBC deformability on cell transport in complex microscale flows. [ABSTRACT FROM AUTHOR]

Altitude training is frequently used by athletes to improve sea-level performance. However, the objective benefits of altitude training are controversial. This study aimed to investigate the possible alterations in hemorheological parameters in response to altitude training. Sprague Dawley rats, were divided into 6 groups: live low-train low (LLTL), live high-train high (LHTH), live high-train low (LHTL) and their controls live high and low (LHALC), live high (LHC), live low (LLC). LHC and LHTH groups were exposed to hypoxia (15% O2, altitudes of 3000 m), 4 weeks. LHALC and LHTL were exposed to 12 hours hypoxia/normoxia per day, 4 weeks. Hypoxia was maintained by a hypoxic tent. The training protocol corresponded to 60-70% of maximal exercise capacity. Rats of training groups ran on treadmill for 20-30 min/day, 4 days/week, 4 weeks. Erythrocyte deformability of LHC group was increased compared to LHALC and LLC. Deformability of LHTH group was higher than LHALC and LLTL groups. No statistically significant alteration in erythrocyte aggregation parameters was observed. There were no significant relationships between RBC deformability and exercise performance. The results of this study show that, living (LHC) and training at altitude (LHTH) seems more advantageous in hemorheological point of view. [ABSTRACT FROM AUTHOR]

Background: The storage of red blood cells (RBC) is associated with impairment of their properties that can induce a circulatory risk to recipients. In a preceding study (2009), we reported that post-storage rejuvenation (RJ) of stored RBC (St-RBC) efficiently reduced the storage-induced RBC/endothelial cell interaction, while only partially reversing the level of intracellular Ca2+, reactive oxygen species, and surface phosphatidylserine. In the present study, we examined the RJ effectiveness in repairing St-RBC mechanical properties. Methods: RBC, stored in CPDA-1 without pre-storage leukoreduction, were subjected to post-storage RJ, and the deformability, osmotic fragility (OF), and mechanical fragility (MF) of the rejuvenated St-RBC (St-RBCRj) were compared to those of untreated St-RBC and of freshly-collected RBC (F-RBC). Results: 5-week storage considerably increased OF and MF, and reduced the deformability of St-RBC. All alterations were only partially (40-70%) reversed by RJ, depending on the extent of the damage: the greater the damage, the lesser the relative effect of RJ. Conclusion: The findings of the present and preceding studies suggest that different St-RBC properties are differentially reversed by RJ, implying that some of the changes occur during storage and are irreversible. © 2014 S. Karger GmbH, Freiburg [ABSTRACT FROM AUTHOR]

This in vitro study investigated time-related effects of propofol at the plasma concentrations required for sedation and general anaesthesia, on RBC aggregation, deformability, and morphology. Blood containing propofol at plasma concentrations of 0, 2 and 4 μg ml-1 was incubated in a water bath at 37°C for 1, 2, or 4 hours. RBC elongation indices (EIs) and aggregation indices (AIs), which represent RBC deformability and aggregation, respectively, were measured. Also, RBC morphological indices (MIs), which represent RBC morphology, were calculated. EIs and AIs were similar at propofol concentrations of 0, 2, or 4 μg ml-1 after 1, 2, or 4 hours of incubation. MIs at propofol plasma concentrations 0 or 2 μg ml-1 were similar after 1, 2, and 4 hours of incubation, however, MI at a propofol concentration of 4 μg ml-1 after 4 hours of incubation was higher than its value after 1 or 2 hours of incubation. No significant difference was observed between MIs at propofol plasma concentrations 0, 2, or 4 μg ml-1 after 1, 2, and 4 hours of incubation. At clinical doses, propofol has no direct effects on RBC deformability, aggregation, or morphology over a 4 hours incubation period. [ABSTRACT FROM AUTHOR]

The present study was designed to investigate the oxidant susceptibility of red blood cells (RBC) from four species (echidna, human, koala, Tasmanian devil) based on changes in cellular deformability. These species were specifically chosen based on differences in lifestyle and/or biology associated with varied levels of oxidative stress. The major focus was the influence of superoxide radicals generated within the cell (phenazine methosulfate, PMS, 50 μM) or in the extracellular medium (xanthine oxidase-hypoxanthine, XO-HX, 0.1 U/ml XO) on RBC deformability at various shear stresses (SS). RBC deformability was assessed by laser-diffraction analysis using a 'slit-flow ektacytometer'. Both superoxide-generating treatments resulted in significant increases of methemoglobin for all species (p<0.01), with Tasmanian devil RBC demonstrating the most sensitivity to either treatment. PMS caused impaired RBC deformability for all species, but vast interspecies variations were observed: human and koala cells exhibited a similar sigmoid-like response to SS, short-beaked echidna values were markedly lower and only increased slightly with SS, while Tasmanian devil RBC were extremely rigid. The effect of XO-HX on RBC deformability was less when compared with PMS (i.e., smaller increase in rigidity) with the exception of Tasmanian devil RBC which exhibited essentially no deformation even at the highest SS; Tasmanian devil RBC response to XO-HX was thus comparable to that observed with PMS. Our findings indicate that ektacytometry can be used to determine the oxidant susceptibility of RBC from different species which varies significantly among mammals representing diverse lifestyles and evolutionary histories. These differences in susceptibility are consistent with species-specific discrepancies between observed and allometrically-predicted life spans and are compatible with the oxidant theory of aging. [ABSTRACT FROM AUTHOR]

In this study, the human RBC capillary flow has been modeled by passing 11 μl of RBC suspension (Hematocrit = 6%) in phosphate buffer solution (PBS) of a viscosity of 1 and 2.6 cP (in the presence of 2% Dextran) through 5 μm pore diameter polycarbonate Nuclepore filters. We have developed a digitally controlled experimental system for measuring the RBC filterability at a constant driving pressure, in the range of 10–400 Pa, producing a wall shear stress range of 1–50 Pa. The RBC filterability was evaluated by measuring the cell suspension flow rate normalized by the PBS flow rate. The RBC filterability has been found to be a nonlinear function of the driving pressure, having a single minimum locus at 25 Pa. Lowering the driving pressure below 25 Pa revealed an unexpected increase of the RBC filterability. The maximal RBC filterability (near unity) was detected at the lowest driving pressure (10 Pa) and the corresponding estimated RBC linear velocity while traveling through the capillary pore was as high as 800 μm/s. Increasing the driving pressure above 25 Pa confirmed previous results, where RBC filterability is monotonically and asymptotically increasing. Increasing the PBS medium viscosity from 1 to 2.26 cP significantly attenuated the RBC filterability and led to the anomalous increase of RBC deformability at the 10 Pa pressure range. We propose that the anomalous increase in RBC deformability was caused by RBCs undergoing spontaneous mechanical fluctuations. [ABSTRACT FROM AUTHOR]

Abstract: Ryanodine receptors (RyRs) are a family of Ca2+ channel proteins that mediate the massive release of Ca2+ from the endoplasmic reticulum into the cytoplasma. In the present study, we manipulated the incorporation of RyR1 into RBC membrane and investigated its influences on the intracellular Ca2+ ([Ca2+]in) level and the biomechanical properties in RBCs. The incorporation of RyR1 into RBC membranes was demonstrated by both immunofluorescent staining and the change of [Ca2+]in of RBCs. In the presence of RyR1, [Ca2+]in showed biphasic changes, i.e., it increased with the extracellular Ca2+ ([Ca2+]ex) up to 5μM and then decreased with the further increase of [Ca2+]ex. However, [Ca2+]in remained constant in the absence of the RyR1. The results of biomechanical measurements on RBCs, including deformability, osmotic fragility, and membrane microviscosity, reflected similar biphasic changes of [Ca2+]in mediated by RyR1 with the increases of [Ca2+]ex. Therefore, it is believed that RyR1 can incorporate into RBC membrane in vitro, and mediate Ca2+ influx, and then regulate RBC biomechanical properties. This information suggests that RBCs may serve as a model to study the function of RyR1 as a Ca2+ release channel. [Copyright &y& Elsevier]

Abstract: Background: Hemorrhagic shock is associated with severe rheological abnormalities. We hypothesized that in the setting of hemorrhagic shock, resuscitation can alter hemorheological characteristics dramatically, and different fluids cause different effects. The aim of this study was to investigate whether the type of fluid administered has an impact on hemorheological characteristics at the early stage of resuscitation in a rodent model of hemorrhagic shock. Methods: Animals were randomized into five groups: (1) sham hemorrhage (SHAM); (2) shock and sham resuscitation (SHOCK); (3) shock and resuscitation with normal saline 32ml/kg (NS); (4) shock and resuscitation with 7.5% hypertonic saline 4ml/kg (HS); (5) shock and resuscitation with 7.5% hypertonic saline/6% Dextran 70 4ml/kg (HSD). Hemorheological characteristics were measured at 60min after resuscitation. Results: Results showed that NS resuscitation deteriorated red blood cell (RBC) deformability compared with the SHOCK group. The HS group showed improved RBC deformability compared with the NS group, although the differences were not statistically significant. There were significant improvements of RBC deformability at all shear rates in the HSD group compared with the NS group. Whole blood and plasma viscosities decreased significantly in the SHOCK group compared with the SHAM group. At shear rates of 60 and 150s−1, the NS group decreased whole blood viscosity compared with the SHOCK group. The HSD group showed elevated plasma viscosity compared with the SHOCK, NS and HS groups. Conclusion: These results suggested that at the early stage of hemorrhagic shock resuscitation, hypertonic–hyperoncotic resuscitation could improve RBC deformability compared with isotonic crystalloid resuscitation. Dextran 70 could elevate plasma viscosity to nearly baseline level. These effects of hypertonic–hyperoncotic resuscitation could be beneficial to maintain microcirculation. [Copyright &y& Elsevier]

Irradiation has been shown to induce biochemical changes in stored red blood cells (RBCs) and to generate reactive oxygen species (ROS). This study evaluated the hemorheological properties, the degree of lipid peroxidation and the oxidative susceptibility of irradiated RBCs. Furthermore, we investigated the radioprotective role of N-t-butyl hydroxylamine (NtBHA) against gamma-ray exposure of RBCs. RBC concentrates were irradiated with a minimum dose of 25 Gy, and were exposed to FeSO4 to examine the oxidative susceptibility. RBC deformability was evaluated by the use of a microfluidic ektacytometer, in relation to the hematological and biochemical properties. The deformability of the irradiated RBCs was significantly lower than that of control. Exposure to gamma rays significantly increased the mean corpuscular volume (MCV) and lipid peroxidation. Changes in RBC deformability were more prominent in irradiated RBCs than in non-irradiated RBCs also under conditions of oxidative stress. The deformability of NtBHA treated RBCs prior to irradiation was not altered as compared with irradiated RBCs not treated with NtBHA. In conclusion, irradiation reduces RBC deformability during storage and the irradiated RBCs seem susceptible to oxidative stress. NtBHA may not have a protective role against the effects of gamma-ray exposure in RBCs but further evaluation of NtBHA or another radioprotective compound is required. [ABSTRACT FROM AUTHOR]

Summary: Objectives: Diabetes mellitus is accompanied by microvascular complications leading to organ dysfunction, while sepsis is a major cause of morbidity and mortality in diabetics. We addressed the hypothesis that red blood cell (RBC) deformability may be additively compromised in septic diabetic patients, leading to a further impairment of microcirculation. Methods: Forty patients suffering from severe sepsis, 12 patients suffering from diabetes and 24 diabetic patients with severe sepsis were enrolled. A filtration method and a hemorheometer were used to measure the RBCs'' index of rigidity (IR). Results: We observed no differences in severity, organ dysfunction and outcome between diabetic and non-diabetic septic patients. Mean SAPS II score was 23.5% vs 26.8% in non-diabetic and diabetic septic patients, respectively. The mortality in non-diabetic septic patients was 22.5% and in septic diabetics was 34.3%, while septic shock occurred in 15.0% and 20.8%, respectively. We detected higher IR (17.72±6.31) in septic diabetics than in patients with diabetes and no sepsis (12.26±2.28, p ≤0.001) and in patients with sepsis and no diabetes (13.9±2.86, p ≤0.01). Conclusion: The presence of diabetes mellitus seems to affect the already compromised RBC deformability of septic patients, probably leading to serious microcirculatory functional impairments in septic diabetic patients. [Copyright &y& Elsevier]

Abstract: Infection-induced RBC dysfunction has been shown to play a role in the modulation of host response to injury and infection. The underlying biochemical mechanisms are not known. This study investigated alterations in RBC band-3 phosphorylation status and its relationship to anion exchange activity in vitro as well as under in vivo septic conditions induced by cecal ligation and puncture (CLP) in mice. Pervanadate treatment in vitro increased band-3 tyrosine phosphorylation that was accompanied by decreased RBC deformability and anion exchange activity. Following sepsis, band-3 tyrosine phosphorylation in whole RBC ghosts as well as in cytoskeleton-bound or soluble RBC protein fractions were elevated as compared to controls. Although anion exchange activity was similar in RBCs from septic and control animals, band-3 interaction with eosin-5-maleimide (EMA), which binds to band-3 lysine moieties, was increased in cells from septic animals as compared to controls, indicating that sepsis altered band 3 organization within the RBC membrane. Since glucose-6-phosphate dehydrogenase is a major antioxidant enzyme in RBC, in order to assess the potential role of oxidative stress in band-3 tyrosine phosphorylation, sepsis-induced RBC responses were also compared between WT and (G6PD) mutant animals (20% of normal G6PD activity). Band-3 membrane content and EMA staining were elevated in G6PD mutant mice compared to WT under control non-septic conditions. Following sepsis, G6PD mutant animals showed lessened responses in band-3 tyrosine phosphorylation and EMA staining compared to WT. RBC anion exchange activity was similar between mutant and WT animals under all tested conditions. In summary, these studies indicate that sepsis results in elevated band-3 tyrosine phosphorylation and alters band-3 membrane organization without grossly affecting RBC anion exchange activity. The observations also suggest that factors other than oxidative stress are responsible for the sepsis-induced increase in RBC band-3 tyrosine phosphorylation. [Copyright &y& Elsevier]

Anemia is the main problem for patients suffering from end stage renal disease (ESRD). This study aimed to determine whether the index of rigidity (IR), that shows red blood cells (RBCs) deformability and the possible IR disturbances can provide an explanation about the cause of anemia, in patients undergoing maintenance hemodialysis (HD) or on peritoneal dialysis. The IR was determined in 39 hemodialyzed patients, who were already in dialysis for a period of time ranging from 16 to 120 months (mean ± SD = 41.8 ± 24.1) (Group A). Furthermore, the IR was measured in 32 patients on continuous ambulatory peritoneal dialysis (CAPD), who were in CAPD for a period of time ranging from 6 to 60 months (mean ± SD = 10.7 ± 9.9) (Group B). Finally, the IR was determined in 17 normal individuals (group C). The RBCs IR was measured twice in group A (before and after the end of a hemodialysis session) and once in groups B and C. The IR was determined by hemorrheometry (method of filtration), using special equipment. In group A the IR was increased in comparison to the control group (C) (17.9 ± 6.2 vs. 10.2 ± 1.8, p < 0.0001). This increase was even higher in the measurement at the end of the hemodialysis session (paired t-test, p < 0.0001). The RBCs IR in CAPD patients was significantly lower than that of HD patients (12 ± 3.8 vs. 17.9 ± 6.2, p < 0.0001) and was not statistically different from the control group (12 ± 3.8 vs. 10.2 ± 1.8, p = 0.068). It is concluded from the study that: 1) in HD patients occur disturbances in the deformability of the RBCs, that are worsened by the hemodialysis session; 2) the index of rigidity of RBCs is significantly higher in the HD patients than in CAPD patients; 3) in patients on CAPD, the disturbance of deformability of the RBCs was less in comparison to the control group, which however does not reach the statistically significant levels. [ABSTRACT FROM AUTHOR]

The hemorheological behavior pre‐ and post‐freezing and tissue survival area (TSA) of frostbitten hind feet in cold‐acclimated rats (group CA) and the control (group C) were observed in order to go further into the mechanism of cold acclimation. The results indicated that blood viscosity and RBC aggregation tendency were lowered and RBC deformability improved obviously in CA rats as compared with those in group C (p<0.01). After the rats' hind feet were frozen, the hemorheological behavior in both group CA and C rats was changed abnormally and the changes were less serious in CA rats than those in C rats. The TSA of CA rats was much larger than that of C rats (p<0.01). The aforesaid results revealed that after rats were acclimated to cold, their resistance to freezing increased and their adaptive changes in hemorheological behavior occurred. The extent of abnormal changes in hemorheological behavior caused by freezing was reduced in CA rats, therefore the disorder microcirculation could be improved. These changes were beneficial to the frostbitten tissue to recovering and healing. So it is considered that in CA rats, the adaptive change in hemorheological behavior is one of the cause enhancing resistance to frostbite. [ABSTRACT FROM AUTHOR]

Introduction: Hereditary spherocytosis (HS) is characterized by decreased erythrocyte deformability resulting in hemolytic anemia. This is a heterogeneous disease regarding underlying protein deficiency, disease severity, age at diagnosis and clinical course. Although largely considered as pediatric disease, HS could be initially diagnosed also in elder patients as a result of gallstones or splenomegaly fortuitous finding. Concurrently, common adulthood metabolic disorders like diabetes or dyslipidemia are also known to impair RBC rheology and deformability. Therefore, we aimed to investigate if these diseases affect the screening and diagnostic tools used for HS diagnosis., Methods: We applied our workflow for HS diagnosis on 95 pathological samples: 29 patients with diabetes, 20 with hypercholesterolemia, 17 with dyslipidemia, 6 with hypertriglyceridemia, 23 with metabolic syndrome (MS). Thus, a total of 73 samples were analyzed by automated reticulocyte analysis, 52 by cryohemolysis test, and 41 by ektacytometry osmoscan analysis as we used two out of the three tests for each individual sample., Results: Applying our screening algorithm based on automated reticulocyte indices, a total of 4 samples (4.2%): one sample (5%) from the diabetes group and three samples (16.7%) from the MS group, positioned into the HS zone. However, no significant difference was found between any of the pathological groups and the controls for the cryohemolysis test or the osmoscan., Conclusion: While diabetes and hypercholesterolemia are pathologic conditions known to present with decreased erythrocyte deformability and disturbed rheology, their possible concomitant presence with HS would not interfere with the screening and confirmatory laboratory methods., (© 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

The influence of isotonic or hypotonic dilution on viscoclastic properties of blood is examined. The viscous, as well as the clastic, properties of blood samples diluted with isotonic saline or pure water, respectively, and of undiluted whole blood samples are compared by means of a dynamic capillary viscosimeter (OCR-D, A. Paar K.G., Austria). The dilution (∼17% of the total blood volume) was performed isovolemically and retained the same rbc count. The rbc swelling observed as a consequence of changes in plasma osmolarity was tracked by the high resolution density measurement, according to the mechanical oscillator technique. Since no significant rbc swelling was found in the dilution with isotonic saline, viscoelastic resistance of blood was efficiently reduced in the observed range of shear rates (2 s-100 s). This decrease is due to reduced plasma protein concentration, which also lowers plasma viscsity by approximately 18%. Although plasma viscosity is significantly decreased in hypotonic dilutions (−12%), tlow properties of the rbc suspensions are, in general, significantly impaired. This is due to the osmotic rbc swelling (hematocrit = +8%), which increases viscous resistance within the suspending fluid, as well as elastic resistance of the rbc due to a loss in rbc deformability. It can be concluded that isotonic dilution leads to a decrease in the viscosity of blood, whereas hypotonic dilution - in an order of magnitude which may occur during resorption of water -Ieads to increased viscous and elastic resistance of the blood. [ABSTRACT FROM AUTHOR]

Red blood cells (RBCs) deformability refers to the cells' ability to adapt their shape to the dynamically changing flow conditions so as to minimize their resistance to flow. The high red cell deformability enables it to pass through small blood vessels and significantly determines erythrocyte survival. Under normal physiological states, the RBCs are attuned to allow for adequate blood flow. However, rigid erythrocytes can disrupt the perfusion of peripheral tissues and directly block microvessels. Therefore, RBC deformability has been recognized as a sensitive indicator of RBC functionality. The loss of deformability, which a change in the cell shape can cause, modification of cell membrane or a shift in cytosol composition, can occur due to various pathological conditions or as a part of normal RBC aging ( in vitro or in vivo ). However, despite extensive research, we still do not fully understand the processes leading to increased cell rigidity under cold storage conditions in a blood bank ( in vitro aging), In the present review, we discuss publications that examined the effect of RBCs' cold storage on their deformability and the biological mechanisms governing this change. We first discuss the change in the deformability of cells during their cold storage. After that, we consider storage-related alterations in RBCs features, which can lead to impaired cell deformation. Finally, we attempt to trace a causal relationship between the observed phenomena and offer recommendations for improving the functionality of stored cells., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Barshtein, Pajic-Lijakovic and Gural.)

In this work we applied a multimodal approach to define the age– and atherosclerosis–related biochemical and functional alterations in red blood cells (RBCs) in ApoE/LDLR−/− mice. Our results revealed that age–related changes in RBCs, such as decreases in RBC deformability and mean height, were more pronounced in ApoE/LDLR−/− mice than in age–matched control mice (C57BL/6J). The decreases in phospholipid content and level of lipid unsaturation were accompanied by an increase in cholesterol esters and esterified lipids in RBC membranes in aged C57BL/6J mice. The age–related decrease in the phospholipid content was more pronounced in ApoE/LDLR−/− mice. In contrast, the increase in the total lipid content in RBC membranes occurred only in ApoE/LDLR−/− mice with advanced atherosclerosis. The age–related alterations also included a decrease in the ratio of turns to α–helices in the secondary structure of hemoglobin (Hb) inside intact RBCs. On the other hand, an increase in the ratio of unordered conformations to α–helices of Hb was observed only in ApoE/LDLR−/− mice and occurred already at the age of 5–weeks. This was related to hypercholesterolemia and resulted in an increased oxygen-carrying capacity. In conclusion, progressive mechanical and functional alterations of RBCs in aged ApoE/LDLR−/− mice were more pronounced than in age–matched C57BL/6J mice. Although, several biochemical changes in RBCs in aged ApoE/LDLR−/− mice recapitulated age–dependent changes observed in control mice, some biochemical features of RBC membranes attributed to hypercholesterolemia were distinct and could contribute to the accelerated deterioration of RBC function in ApoE/LDLR−/− mice. Unlabelled Image • Age–related mechanical and functional changes of RBCs are accelerated in ApoE/LDLR−/− mice. • Advanced atherosclerosis is linked with an increase in total lipid content in RBC membranes. • Aging and hypercholesterolemia progression affect secondary structure of Hb differently. • Hypercholesterolemia in young ApoE/LDLR−/− mice induces RBC changes prior to atherosclerosis. [ABSTRACT FROM AUTHOR]

In blood banking practice, the storage duration is used as the primary criterion for inventory management, and usually, the packed red blood cells (PRBC) units are supplied primarily according to first-in-first-out (FIFO) principle. However, the actual functionality of individual PRBC units is mostly ignored. One of the main features of the RBCs not accounted for under this approach is the deformability of the red cells, i.e., their ability to affect the recipients' blood flow. The objective of the study was to analyze unit-to-unit variability in the deformability of PRBCs during their cold storage. RBC samples were obtained from twenty leukoreduced PRBC units, stored in SAGM. The deformability of cells was monitored from the day of donation throughout 42 days. RBC deformability was determined using the computerized cell flow-properties analyzer (CFA) based on cell elongation under a shear stress of 3.0 Pa, expressed by the elongation-ratio (ER). The image analysis determines the ER for each cell and provides the ER distribution in the population of 3000–6000 cells. The deformability of freshly-collected RBCs exhibited marked variability already on the day of donation. We also found that the aging curve of PRBC deformability varies significantly among donors. The present study has demonstrated that storage duration is only one of the factors, and seemingly not even the major one, affecting the PRBCs functionality. Therefore, the FIFO approach is not sufficient for assessing the potential transfusion outcome, and the PRBC functionality should be determined explicitly for each unit. [ABSTRACT FROM AUTHOR]

Throughout storage, red blood cells (RBCs) undergo detrimental changes in viability and their ability to effectively transport oxygen. RBC storage lesions are mediated, in part, by a progressive loss of cell deformability, and associated with the release of extracellular vesicles (EVs). Accumulation of EVs during the storage of RBCs correlates with a decrease in RBC surface area to volume ratio. Similarly, the loss of RBC-deformability is associated with loss of RBC surface area to volume ratio. In this study we thus tested whether loss of RBC-deformability is associated with increased RBC-EV production during blood storage. EVs obtained by differential centrifugation of stored RBCs (non-leukoreduced non-irradiated or leukoreduced γ-irradiated RBCs stored 35 or 28 days respectively) were enumerated by high-sensitivity flow cytometry. RBC deformability was quantified, using a cell-flow-properties-analyzer, by measuring the median cell elongation ratio (MER) and percentage of low and high deformable cells in the population (%, LDFC, and HDFC, respectively). The number of EVs was inversely correlated with the MER and positively correlated with the %LDFC with both measures showing highly significant logarithmic dependence with EV levels in stored RBCs. Considering how highly deformable cells did not correlate with EV formation as compared with low deformable RBCs we propose that the formation of EVs is a key factor leading to increased RBC-rigidity. [ABSTRACT FROM AUTHOR]

Donated blood is stored in the blood bank as packed red blood cell units. In the process of packed cells preparation, the red blood cells (RBCs) are subjectedto high level of shear stress, which can induce alterations in their properties. In the present study, we examined the effect of packed RBCs preparation (which included leuko-filtration) on red cell deformability. Blood samples were collected from 25 healthy donors and from corresponding units of packed RBCs. The portion of undeformable cells (%UDFC) was determined for each sample. The median value of %UDFC was equal to 6.75 ± 0.70 %, for freshly-donated RBCs, and to 6.36 ± 0.51 %, for packed cells. Wherein, %UDFC may increase or decrease following packed cells preparation, depending upon the initial portion of undeformable cells. Likely, exposure of RBCs to high shear stress, during packed cells preparation, induces opposing effects: (a) removal/destruction of rigid (undeformable) cells, thereby reducing their total amount (i.e., decreasing the %UDFC) on the one hand, and (b) mechanical damage to the cell membrane and subsequent reduction of the cell deformability (thereby increasing the %UDFC) on the other. As a consequence, the final impact of packed cells preparation is primarily determined by the initial state of erythrocytes in the blood of the donor. [ABSTRACT FROM AUTHOR]

Aim: Nitric oxide (NO) prevents the decline of RBC deformability under high altitude and other ischemic and hypoxic conditions, but the clear mechanisms remain unknown. Here, we have carried out a systematic study to find the mechanisms of NO-induced regulation of RBC deformability under hypoxia., Methods: NO levels, RBCs membrane elongation index (EI), membrane protein band 3 methemoglobin (MetHb) were determined during hypoxia (0 to 120 minutes). To validate the role of NO in regulating RBC deformability, tests were also performed with a NO donor (sodium nitroprusside) or a NO synthase inhibitor (l-nitro-arginine methylester) under 60 minutes hypoxia., Results: Hypoxia for 45 minutes increased NO levels from 25.65 ± 1.95 to 35.26 ± 2.01 μmol/L, and there was a plateau after 60 minutes hypoxia. The EI did not change before 45 minutes hypoxia, but decreased from 0.567 ± 0.019 to 0.409 ± 0.042 (30 Pa) after 60 minutes hypoxia. The cross-linking of band 3 and phosphotyrosine increased after 45 minutes hypoxia. All can be alleviated by supplement NO and aggregated by inhibiting NOS. However, the MetHb was not present this trend., Conclusion: NO may prevent decreased of RBCs deformability through reducing the cross-linking of membrane band 3 under hypoxia; this helps microvascular perfusion of RBCs during ischemic and hypoxic disease states., (© 2018 Wiley Periodicals, Inc.)

Background: The Nordic hamstring exercise (NHE) has been proven to be an effective preventive technique for hamstring injuries. Hemorheological parameters (erythrocyte deformability and aggregation) play a critical role in exercise influencing oxygenation. Although previous studies presented hemorheological alterations induced by different types of exercise, changes in red blood cell (RBC) deformability and aggregation following NHE remain unknown. Present study was designed to explore possible alterations in hemorheological and oxidative parameters after an acute bout of NHE., Methods: 10 healthy, male, active students (mean age 19.9±0.23, BMI: 21.56±0.54) participated to the study. They performed a single session of seven-repetitions of NHE followed by a familiarisation period. Blood samples were obtained before and immediately after the exercise from the antecubital vein. Hemorheological parameters were measured by an ektacytometer., Results: NHE did not change deformability, hematocrit and oxidative stress but, increased RBC aggregation index (AI, p = 0.011) and decreased RBC aggregation half time (t½, p = 0.009)., Conclusions: Our results suggest that, increased RBC aggregation following an acute bout of NHE may result in increased plasma skimming and thus ease the flow of blood.

Biophysical properties are widely used to detect pathophysiological processes of vascular diseases or clinical states. For early detection of cardiovascular diseases, it is necessary to simultaneously measure multiple biophysical properties in a microfluidic environment. However, a microfluidic-based technique for measuring multiple biophysical properties has not been demonstrated. In this study, a simple measurement method was suggested to quantify three biophysical properties of blood, including red blood cell (RBC) deformability, RBC aggregation, and hematocrit. To demonstrate the suggested method, a microfluidic device was constructed, being composed of a big-sized channel (BC), a parallel micropillar (MP), a main channel, a branch channel, inlet, and outlets. By operating a single syringe pump, blood was supplied into the inlet of the microfluidic device, at a periodic on-off profile (i.e., period = 240 s). The RBC deformability index (DI) was obtained by analyzing the averaged blood velocity in the branch channel. Additionally, the RBC aggregation index (AIN) and the hematocrit index (HiBC) were measured by analyzing the image intensity of blood flows in the MP and the BC, respectively. The corresponding contributions of three influencing factors, including the turn-on time (Ton), the amplitude of blood flow rate (Q0), and the hematocrit (Hct) on the biophysical indices (DI, AIN, and HiBC) were evaluated quantitatively. As the three biophysical indices varied significantly with respect to the three factors, the following conditions (i.e., Ton = 210 s, Q0 = 1 mL/h, and Hct = 50%) were maintained for consistent measurement of biophysical properties. The proposed method was employed to detect variations of biophysical properties depending on the concentrations of autologous plasma, homogeneous hardened RBCs, and heterogeneous hardened RBCs. Based on the observations, the proposed method exhibited significant differences in biophysical properties depending on base solutions, homogeneous hardened RBCs (i.e., all RBCs fixed with the same concentration of glutaraldehyde solution), and heterogeneous hardened RBCs (i.e., partially mixed with normal RBCs and homogeneous hardened RBCs). Additionally, the suggested indices (i.e., DI, AIN, and HiBC) were effectively employed to quantify three biophysical properties, including RBC deformability, RBC aggregation, and hematocrit. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate changes in RBC aggregation and deformability over 24 hr and suggest specific shear stress values for detecting RBC deformability in a mouse endotoxemia model using lipopolysaccharide (LPS).Six-week-old male BALB/c mice received LPS (20 mg/kg) intraperitoneally. Aggregation indices (AIs) and T1/2 were measured to assess RBC aggregation, and elongation indices (EIs) were used to assess RBC deformability at shear stress values of 0.3, 0.5, 1, 3, 7, 10, 15 and 20 Pascals (Pa) 0, 30 min, 1, 2, 4, 6, 9, 12, 18 and 24 hours after the LPS injection.No significant differences were detected in the AIs during the study period, however, T1/2 shortened significantly 2, 6, 12, 18, and 24 hr after the LPS injection. The EIs increased significantly 24 hr after LPS injection at 0.5 and 1 Pa shear stress, whereas it decreased significantly at 10 Pa of shear stress 24 hr after the LPS injection.Altered RBC deformability was detectable 24 hr after the LPS injection and T1/2 may be a sensitive marker for detecting changes in RBC aggregation. The EIs should be measured at 1 Pa to detect changes in RBC deformability in LPS-induced septic mice.

Even though exercise hemorheology at hypoxic condition has been considered as a good tool to understand clinical hemorheology, there have been limited studies reported. Previous researches showed that hemorheological variables are closely correlated with oxygen delivery capacity during exercise. The present study investigated hypoxic responses including RBC deformability and aggregation, metabolic parameters and complete blood cell counts at various hypoxic conditions during cycling exercise and recovery. Eleven Korean healthy male subjects performed submaximal bike exercise at sea level (20.9% O2) and under various hypoxic conditions (16.5% O2, 14.5% O2, 12.8% O2, and 11.2% O2) in a random order. The submaximal bike exercise intensity of the subjects was 70% maximum heart rate at sea level. All variables were measured at rest, during exercise and recovery 30-minute, respectively. As oxygen partial pressure decreased, arterial blood oxygen saturation decreased but oxygen uptake did not change much. Heart rate and lactate concentration during exercise increased when oxygen partial pressure is less than or equal to 14.5% O2 condition. Red blood cell (RBC) counts, hemoglobin counts, and hematocrit level were not apparently altered with hypoxic conditions. RBC deformability showed significant alterations at 11.2% O2 conditions compared with other hypoxic conditions during exercise or recovery, except at 10 minutes recovery. However, decreases in oxygen partial pressure did not affect red blood cell aggregation. Therefore, we conclude that alterations in RBC deformability may reduce aerobic capabilities at hypoxic condition.