Asunto(s)
Eritrocitos/fisiología , Hematología/historia , Microscopía/historia , Agregación Eritrocitaria/fisiología , Deformación Eritrocítica/fisiología , Eritrocitos/citología , Historia del Siglo XVII , Historia del Siglo XVIII , Historia del Siglo XIX , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Reología/historiaRESUMEN
BACKGROUND: The recent in vitro demonstration that inositol hexaphosphate-loaded red blood cells (IHP-RBCs) may reduce the risks of sickling of sickle RBCs (SS RBCs) exposed to hypoxia make these modified RBCs potentially useful in transfused sickle cell anemia (SCA) patients. STUDY DESIGN AND METHODS: Hemorheologic properties of IHP-RBCs, normal RBCs (AA RBCs), SS RBCs, SS RBCs plus AA RBCs, and SS RBCs plus IHP-RBCs were compared under normoxia and/or after hypoxic challenges. RESULTS: Although IHP-RBCs have reduced deformability compared with SS RBCs or AA RBCs, IHP-RBCs exhibited lower aggregability than AA RBCs and SS RBCs and, when mixed with SS RBCs, the aggregation level was below the one of SS RBCs alone or SS RBCs plus AA RBCs. Blood viscosity of SS RBC plus IHP-RBC suspension was lower than the viscosity of SS RBCs alone and greater than viscosity of SS RBCs plus AA RBCs. The hypoxic challenge was detrimental for deformability and viscosity of SS RBCs alone or SS plus AA RBC suspension but not for SS plus IHP-RBC suspension. CONCLUSION: Our results support the fact that IHP-RBCs could be useful in SCA by decreasing RBC aggregation and blunting the adverse effects of hypoxia on RBC deformability and blood viscosity.
Asunto(s)
Anemia de Células Falciformes/sangre , Eritrocitos Anormales/efectos de los fármacos , Eritrocitos/fisiología , Hemorreología/efectos de los fármacos , Ácido Fítico/farmacología , Anemia de Células Falciformes/patología , Viscosidad Sanguínea/efectos de los fármacos , Agregación Eritrocitaria/efectos de los fármacos , Eritrocitos/efectos de los fármacos , Eritrocitos/patología , Eritrocitos Anormales/patología , Humanos , Fragilidad Osmótica/efectos de los fármacos , Ácido Fítico/administración & dosificación , Resistencia al Corte/efectos de los fármacos , Estrés MecánicoRESUMEN
AIMS: Distorted wall shear stress (WSS) in patients with type 2 diabetes mellitus (T2DM) may be partly explained by an altered red blood cell aggregation tendency (RAT) on viscosity at low shear rate (SR). The present study evaluates viscosity modeling by implementation of hematocrit and RAT in patients with and without T2DM (non-T2DM). METHODS: A Couette viscometer and LORCA aggregometer provided viscosity and RAT on 6 shear rates in 55 patients (46-78 yrs, 66% male, T2DM: n = 28), following informed consent. Using a K-fold cross-validation, two linear mixed models predicted by SR and Hct and by SR, Hct and RAT were compared. RESULTS: In non-T2DM modeling was improved in relatively low RATs (48%, p = 1.0 x 10-11) and became worse in relatively high RATs (-18%, p = 0.019). In T2DM the opposite was observed, as modeling became worse in relatively low RATs (-16%, p = 0.001) but was improved in relatively high RATs (22%, p = 0.022). CONCLUSIONS: In addition to confirming previous research, major differences in modeling improvement between T2DM and non-T2DM were found. Especially patients with T2DM, a high RAT and often high viscosity at low SR benefit from a more accurate viscosity modeling. Further studies should evaluate how these findings affect WSS in these patients.
Asunto(s)
Viscosidad Sanguínea , Diabetes Mellitus Tipo 2/sangre , Agregación Eritrocitaria , Estrés Mecánico , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos BiológicosRESUMEN
BACKGROUND: Test 1 is a recently introduced technique claiming to determine Erythrocyte Sedimentation Rate (ESR) in 20 s. In contrast to the original Westergren procedure this new technique uses undiluted blood and operates at 37 degrees C. It is hypothesized that Test 1 is in fact an erythrocyte aggregometer and does not measure any sedimentation. METHODS: Test 1 results were compared to those obtained with StaRRsed, an automated ESR analyser based on the Westergren technique, and the results of both were correlated to various indices of red blood cell (RBC) aggregation, obtained with an aggrego--meter (LORCA). Measurements were made on blood from 75 patients with various rheumatic disorders. Furthermore, blood that was experimentally manipulated in order to affect RBC aggregation, i.e. by changing the hematocrit, by diminishing plasma protein concentration, by inducing hyperaggregation or by RBC rigidification, was tested on all three instruments. RESULTS: Generally in patient blood, Test 1 results demonstrated a higher correlation with the various aggregation parameters than StaRRsed. Highest correlation (R = -0.8)) with both Test 1 and StaRRsed outcome were seen with I(20), a RBC aggregation parameter directly related to the backscatter intensity. All experimentally induced changes in RBC aggregation paralleled closely those obtained with Test 1 while StaRRsed results followed a different course. CONCLUSIONS: The results obtained in this study strongly support the hypothesis that Test 1 measures only the RBC aggregation process and does not cover any of the indices directly linked to the sedimentation process as determined by the Westergren method.
Asunto(s)
Sedimentación Sanguínea , Agregación Eritrocitaria , Pruebas Hematológicas/instrumentación , Autoanálisis/instrumentación , Autoanálisis/métodos , Proteínas Sanguíneas/análisis , Viscosidad Sanguínea , Deformación Eritrocítica , Hematócrito , Pruebas Hematológicas/normas , Humanos , Enfermedades Reumáticas/sangreRESUMEN
BACKGROUND: various modifications of the Erythrocyte Sedimentation Rate (ESR) determination have been suggested since the original Westergren procedure that has been adopted as the gold standard by the International Council for Standardization in Haematology (ICSH). Recently, an automated method, (Alifax Test 1), based on a technique completely different from Westergren, has been introduced. MATERIAL AND METHODS: In this comparative study, ESR of blood from 680 patients with various rheumatic diseases was determined on both Test 1 and the StaRRsed automated ESR analyser which performs measurements in accordance with ICSH specifications. Furthermore the robustness of the new technique was evaluated. RESULTS: Direct correlation of Test 1 and StaRRsed measurements confirmed the results of previous studies: an overall correlation coefficient of R = 0.90. However, further statistical analysis showed that, depending on the instrument that was used, in 78 samples (i.e. 11.5%) the results could lead to different treatment suggestions. Furthermore it appeared that several procedural factors could influence the final Test 1 outcome. CONCLUSIONS: Due to its sensitivity for procedural variations, Test 1 measurements should be carried out under strictly standardized conditions. Especially at the higher ESR levels the Test 1 technique is, however, not a reliable alternative for the ICSH approved 'Westergren' method.
Asunto(s)
Sedimentación Sanguínea , Pruebas Hematológicas/instrumentación , Automatización de Laboratorios/instrumentación , Pruebas Hematológicas/métodos , Humanos , Estándares de Referencia , Enfermedades Reumáticas/sangre , Sensibilidad y EspecificidadRESUMEN
The International Society for Clinical Hemorheology organized a workshop to compare three instruments for measuring RBC aggregation: LORCA, Myrenne Aggregometer and RheoScan-A. The Myrenne Aggregometer provides indices at stasis (M) and at low shear (M1), with four indices obtained with the LORCA and RheoScan-A: amplitude (AMP), half-time (T1/2), surface area (SA) above (LORCA) or below (RheoScan-A) the syllectogram, and the ratio (AI) of the area above (LORCA) or below (RheoScan-A) the syllectogram to total area (AI). Intra-assay reproducibility and biological variability were determined; also studied were RBC in diluted plasma and in 1% 500 kDa dextran, and 0.003% glutaradehyde (GA)-treated cells in plasma. All measurements were performed at 37 degrees C. Standardized difference values were used as a measure of power to detect differences. Salient results were: (1) intra-assay variations below 5% except for RheoScan-A AMP and SA; (2) biological variability greatest for T1/2 with other indices similar for the three devices; (3) all instruments detected progressive changes with plasma dilution; (4) the Myrenne and LORCA, but not the RheoScan-A, detected differences for cells in dextran; (5) GA-treatment significantly affected the LORCA (AMP, T1/2, SA, AI), the RheoScan-A (AMP, SA, AI) and the Myrenne M parameter. It is concluded that: (a) the LORCA, Myrenne and the RheoScan-A have acceptable precision and suitable power for detecting reduced aggregation due to plasma dilution; (b) greatly enhanced RBC aggregation may not be sensed by the RheoScan-A while the Myrenne M1 index may be insensitive to minor increases of cell rigidity; (c) future studies should define each instrument's useful range for detecting RBC aggregation.
Asunto(s)
Agregación Eritrocitaria/fisiología , Eritrocitos/fisiología , Reología/instrumentación , Adulto , Sedimentación Sanguínea , Deformación Eritrocítica/fisiología , Hemorreología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Red blood cell (RBC) aggregation is the reversible and regular clumping in the presence of certain macromolecules. This is a clinically important phenomenon, being significantly enhanced in the presence of acute phase reactants (e.g., fibrinogen). Both light reflection (LR) and light transmission (LT) from or through thin layers of RBC suspensions during the process of aggregation are accepted to reflect the time course of aggregation. It has been recognized that the time courses of LR and LT might be different from each other. We aim to compare the RBC aggregation measurements based on simultaneous recordings of LR and LT. The results indicate that LR during RBC aggregation is characterized by a faster time course compared to simultaneously recorded LT. This difference in time course of LR and LT is reflected in the calculated parameters reflecting the overall extent and kinetics of RBC aggregation. Additionally, the power of parameters calculated using LR and LT time courses in detecting a given difference in aggregation are significantly different from each other. These differences should be taken into account in selecting the appropriate calculated parameters for analyzing LR or LT time courses for the assessment of RBC aggregation.
Asunto(s)
Algoritmos , Agregación Eritrocitaria/fisiología , Eritrocitos/citología , Eritrocitos/fisiología , Fotometría/métodos , Refractometría/métodos , Adulto , Células Cultivadas , Humanos , Luz , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Dispersión de Radiación , Sensibilidad y EspecificidadRESUMEN
Measurement of red blood cell (RBC) deformability by ektacytometry yields a set of elongation indexes (EI) measured at various shear stresses (SS) presented as SS-EI curves, or tabulated data. These are useful for detailed analysis, but may not be appropriate when a simple comparison of a global parameter between groups is required. Based on the characteristic shape of SS-EI curves, two approaches have been proposed to calculate the maximal RBC elongation index (EI(max)) and the shear stress required for one-half of this maximal deformation (SS(1/2)): (i) linear Lineweaver-Burke (LB) model; (ii) Streekstra-Bronkhorst (SB) model. Both approaches have specific assumptions and thus may be subject to the measurement conditions. Using RBC treated with various concentrations of glutaraldehyde (GA) and data obtained by ektacytometry, the two approaches have been compared for nine different ranges of SS between 0.6-75 Pa. Our results indicate that: (i) the sensitivity of both models can be affected by the SS range and limits employed; (ii) over the entire range of SS-data, a non-linear curve fitting approach to the LB model gave more consistent results than a linear approach; (iii) the LB method is better for detecting SS(1/2) differences between RBC treated with 0.001-0.005% glutaraldehyde (GA) and for a 40% mixture of rigid cells but is equally sensitive to SB for 10% rigid cells; and (iv) the LB and SB methods for EI(max) are equivalent for 0.001% and 0.003% GA and 40% rigid, with the SB better for 0.005% GA and the LB better for 10% rigid.
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Técnicas Citológicas/métodos , Deformación Eritrocítica/fisiología , Eritrocitos/fisiología , Estrés Mecánico , Adulto , Anciano , Deformación Eritrocítica/efectos de los fármacos , Eritrocitos/efectos de los fármacos , Glutaral/farmacología , Humanos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Dinámicas no Lineales , Estándares de Referencia , Análisis de RegresiónRESUMEN
Rheological studies concerning aggregation and elongation of erythrocytes were carried out in 21 patients (mean age 56 years) with chronic venous disease (CVD) and 10 (mean age 45 years) healthy control subjects, with the use of a LORCA device. Higher values of parameters characterizing both erythrocyte elongation (EI) and aggregation (gammathr) in non-control patients than in the control group were found. These values differed significantly ranging from 1.13 to 8.23 Pa for the shear stress and gammathr in patients--432.14, in relation to the control group--166.5. It was proposed, that the increase in deformability may constitute a compensatory mechanism in subjects with chronic venous disease, due to increased resistance in their microcirculation.
Asunto(s)
Deformación Eritrocítica , Eritrocitos/metabolismo , Insuficiencia Venosa/sangre , Adolescente , Adulto , Anciano , Agregación Celular , Enfermedad Crónica , Eritrocitos/patología , Femenino , Humanos , Masculino , Microcirculación , Persona de Mediana Edad , Insuficiencia Venosa/patología , Insuficiencia Venosa/fisiopatologíaRESUMEN
BACKGROUND: Hypertension is associated with inward remodeling of small arteries and decreased erythrocyte deformability, both impairing proper tissue perfusion. We hypothesized that these alterations depend on transglutaminases, cross-linking enzymes present in the vascular wall, monocytes/macrophages and erythrocytes. METHODS AND RESULTS: Wild-type (WT) mice and tissue-type transglutaminase (tTG) knockout (KO) mice received the nitric oxide inhibitor Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME) to induce hypertension. After 1 week, mesenteric arteries from hypertensive WT mice showed a smaller lumen diameter (-6.9 +/- 2.0%, p = 0.024) and a larger wall-to-lumen ratio (11.8 +/- 3.5%, p = 0.012) than controls, whereas inward remodeling was absent in hypertensive tTG KO mice. After 3 weeks, the wall-to-lumen ratio was increased in WT (20.8 +/- 4.8%, p = 0.005) but less so in tTG KO mice (11.7 +/- 4.6%, p = 0.026), and wall stress was normalized in WT but not in tTG KO mice. L-NAME did not influence expression of tTG or an alternative transglutaminase, coagulation factor XIII (FXIII). Suppression of FXIII by macrophage depletion was associated with increased tTG in the presence of L-NAME. L-NAME treatment decreased erythrocyte deformability in the WT mice (-15.3% at 30 dynes/cm(2), p = 0.014) but not in the tTG KO mice. CONCLUSION: Transglutaminases are involved in small artery inward remodeling and erythrocyte stiffening associated with nitric oxide inhibition-related hypertension.
Asunto(s)
Deformación Eritrocítica , Proteínas de Unión al GTP/metabolismo , Hipertensión/metabolismo , Arterias Mesentéricas/metabolismo , Óxido Nítrico/metabolismo , Transglutaminasas/metabolismo , Animales , Presión Sanguínea , Ácido Clodrónico/farmacología , Modelos Animales de Enfermedad , Inhibidores Enzimáticos , Factor XIII/metabolismo , Proteínas de Unión al GTP/deficiencia , Proteínas de Unión al GTP/genética , Hematócrito , Hipertensión/sangre , Hipertensión/inducido químicamente , Hipertensión/enzimología , Hipertensión/patología , Hipertensión/fisiopatología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Arterias Mesentéricas/enzimología , Arterias Mesentéricas/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , NG-Nitroarginina Metil Éster , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/metabolismo , Proteína Glutamina Gamma Glutamiltransferasa 2 , Proyectos de Investigación , Estrés Mecánico , Factores de Tiempo , Transglutaminasas/deficiencia , Transglutaminasas/genéticaRESUMEN
The effect of elevated shear stress upon cellular trauma has been studied for many years, but the effect of long-term cyclic stress trauma on hemorheology has never been explored systematically. This study investigated sublytic trauma of red blood cells (RBCs) caused by repeated exposure to shear stress. A suspension of bovine blood was throttled through a capillary tube (inner diameter 1 mm and length 70 mm) connected to a recirculating flow loop. Samples were withdrawn every 30 min to measure deformability and characteristic time. The deformability of the cell was measured microscopically by observing the shape of the cell during the shear flow. It was found that cyclic shear irreversibly stiffened the cell membrane while the effect was not so much as that of continuous shear. The cell deformability was dramatically reduced by 73% when the stress of 300 Pa was applied for 288 s, while it was 7% under 90 Pa. These results elucidate the need for improved models to predict cellular trauma within the unsteady flow environment of mechanical circulatory assist devices.
Asunto(s)
Deformación Eritrocítica/fisiología , Eritrocitos/citología , Eritrocitos/fisiología , Animales , Bovinos , Hemólisis , Hemorreología/métodos , Resistencia al Corte , Estrés MecánicoRESUMEN
OBJECTIVE: Scavenger receptor class B, type I (SR-BI) is a multifunctional receptor that promotes the selective uptake of cholesteryl esters from high-density lipoprotein (HDL). Disruption of SR-BI in mice results in a dramatic increase in HDL cholesterol. Interestingly, mice lacking SR-BI also develop anemia, as evidenced by accumulation of reticulocytes in the circulation. The objective of the current study was to delineate the mechanism underlying development of anemia in the absence of SR-BI. METHODS: Expression of important mediators of erythropoiesis, as well as key enzymes in the degradation of erythrocytes, were analyzed using real-time polymerase chain reaction in SR-BI wild-type and SR-BI knockout mice. In addition, in vivo studies were performed using biotinylated erythrocytes to determine erythrocyte survival. RESULTS: mRNA expression of TAL-1, GATA-1, FOG-1, erythropoietin receptor, and ferrochelatase, important mediators of erythropoiesis, was increased in spleens of SR-BI-deficient mice. In addition, the relative amount of early Ter119(high)CD71(high) -expressing erythroblasts was increased in SR-BI-deficient spleens. Interestingly, also expression of hemeoxygenase 1 and biliverdin reductase, enzymes involved in the degradation of erythrocytes, was increased. Furthermore, an elevated amount of conjugated bilirubin, the breakdown product of hemoglobin, was found in bile. Using biotinylated erythrocytes, we show that survival of erythrocytes was decreased in SR-BI-deficient mice. Thus, the observed increased erythropoiesis in the SR-BI-deficient mice is most likely a direct response to the reduced erythrocyte lifespan. Finally, we show that increased HDL cholesterol levels due to SR-BI deficiency induce erythrocyte cholesterol:phospholipid ratios, resulting in decreased deformability and increased osmotic fragility, thereby providing an explanation for the observed reduced lifespan. CONCLUSIONS: SR-BI is not only essential for HDL cholesterol homeostasis and atherosclerosis susceptibility, but also for maintaining normal erythrocyte lifespan.
Asunto(s)
HDL-Colesterol/sangre , Envejecimiento Eritrocítico/genética , Anemia/etiología , Animales , Colesterol/análisis , HDL-Colesterol/fisiología , Deformación Eritrocítica , Eritropoyesis/genética , Perfilación de la Expresión Génica , Hemo-Oxigenasa 1/genética , Ratones , Ratones Noqueados , Fragilidad Osmótica , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/genética , Fosfolípidos/análisis , ARN Mensajero/análisis , Receptores Depuradores de Clase B/deficiencia , Bazo/químicaRESUMEN
BACKGROUND: The ability to deform is important for circulating RBCs in vivo, and earlier studies showed that this property can objectively be measured in vitro by the LORCA. In this study it was investigated whether photodynamic treatment of human RBCs (meant to inactivate contaminating pathogens) affects deformability. STUDY DESIGN AND METHODS: WBC-reduced RBC suspensions (30% Hct) were treated with 1,9-dimethylmethylene blue (DMMB) and red light. Changes in deformability were analyzed by LORCA measurements, in which elongation of the cells is measured at increasing shear stress. The effect of DMMB concentration and light dose was determined as well as the interfering effect of two scavengers of reactive oxygen species, that is, dipyridamole and Trolox. RESULTS: Photodynamic treatment with DMMB resulted in clear changes in RBC deformability. Deformability changes occurred before onset of hemolysis. Under relatively mild treatment conditions, especially deformability at low shear stress was decreased, whereas deformability changes at high shear stress only occurred under harsher treatment conditions. Inclusion of dipyridamole and/or Trolox primarily prevented deformability changes at high shear stress. CONCLUSION: LORCA measurements can effectively be used to detect changes in deformability that are induced by photodynamic treatment of human RBCs. A change in deformability represents an early marker of RBC damage under these conditions.