Which sub-compartments of fat mass and fat-free mass are related to blood viscosity factors?

The size of body compartments is a determinant of several factors of blood viscosity. Red cell aggregation is proportional to fat mass while hematocrit is proportional to both fat-free mass and abdominal adiposity, but which parts of these body components are involved in this relationship is not known. Segmental bioelectrical impedance analysis (sBIA) provides a possibility to delineate the relationships more precisely between various subdivisions of the body and blood viscosity factors, going farther than preceding studies using non segmental BIA. In this study we investigated in 38 subjects undergoing a standardized breakfast test with mathematical modelling of glucose homeostasis and a segmental bioelectrical impedance analysis (sBIA) the relationships between the various compartments of the body and viscosity factors. Blood and plasma viscosity were measured with the Anton Paar rheometer and analyzed with Quemada's model. The parameters better correlated to hematocrit are fat free mass (r = 0.562) and its two components muscle mass (r = 0.516) and non-muscular fat-free mass (r = 0.452), and also trunk fat mass (r = 0.383) and waist-to hip ratio (r = 0.394). Red cell aggregation measurements were correlated with both truncal and appendicular fat mass (r ranging between 0.603 and 0.728). Weaker correlations of M and M1 are found with waist circumference and hip circumference. This study shows that the correlation between lean mass and hematocrit involves both muscle and non-muscle moieties of lean mass, and that both central and appendicular fat are determinants of red cell aggregation.

Novel blood typing method by discrimination of hemagglutination and rouleaux using an erythrocyte aggregometer.

BACKGROUND: In pretransfusion blood typing, pretreatments such as centrifugation and suspension of red blood cells (RBCs) and mixing them with sufficient amounts of reagents are required, but these steps are time-consuming and costly. OBJECTIVE: Aiming to develop a new blood typing method that requires no dilution and only a small amount of reagent, we attempted to determine blood type using syllectometry, an easy-to-use and rapid optical method for measuring the RBC aggregation that occurs when blood flow is abruptly stopped in a flow channel. METHODS: Samples of whole blood from 20 healthy participants were mixed with antibody reagents for blood typing at mixing ratios of 2.5% to 10% and measured with a syllectometry device. RESULTS: Amplitude (AMP), one of the aggregation parameters, showed significant differences between agglutination and non-agglutination samples at mixing ratios from 2.5% to 10%. Although there were significant individual differences in aggregation parameters, calculation of AMP relative to that of blood before reagent mixing reduced the individual differences and enabled determination of blood type in all participants. CONCLUSIONS: This new method enables blood typing with a small amount of reagent, without the time-consuming and labor-intensive pretreatments such as centrifugation and suspension of RBCs.

Axial shear rate: A hemorheological factor for erythrocyte aggregation under Womersley flow in an elastic vessel based on numerical simulation.

Erythrocyte aggregation (EA) is a highly dynamic, vital phenomenon to interpreting human hemorheology, which would be helpful for the diagnosis and prediction of circulatory anomalies. Previous studies of EA on erythrocyte migration and the Fåhraeus Effect are based on the microvasculature. They have not considered the natural pulsatility of the blood flow or large vessels and mainly focused on shear rate along radial direction under steady flow to comprehend the dynamic properties of EA. To our knowledge, the rheological characteristics of non-Newtonian fluids under Womersley flow have not reflected the spatiotemporal behaviors of EA or the distribution of erythrocyte dynamics (ED). Hence, it needs to interpret the ED affected by temporal and spatial flow variation to understand the effect of EA under Womersley flow. Here, we demonstrated the numerically simulated ED to decipher EA's rheological role in axial shear rate under Womersley flow. In the present study, the temporal and spatial variations of the local EA were found to mainly depend on the axial shear rate under Womersley flow in an elastic vessel, while mean EA decreased with radial shear rate. The localized distribution of parabolic or M-shape clustered EA was found in a range of the axial shear rate profile (-15 to 15s-1) at low radial shear rates during a pulsatile cycle. However, the linear formation of rouleaux was realized without local clusters in a rigid wall where the axial shear rate is zero. In vivo, the axial shear rate is usually considered insignificant, especially in straight arteries, but it has a great impact on the disturbed blood flow due to the geometrical properties, such as bifurcations, stenosis, aneurysm, and the cyclic variation of pressure. Our findings regarding axial shear rate provide new insight into the local dynamic distribution of EA, which is a critical player in blood viscosity. These will provide a basis for the computer-aided diagnosis of hemodynamic-based cardiovascular diseases by decreasing the uncertainty in the pulsatile flow calculation.

Relationship between red blood cell aggregation and dextran molecular mass.

The aim of this study was to investigate the aggregation of red blood cells (RBCs) suspended in dextran solution at various levels of molecular mass. Dextran solutions at molecular mass 40, 70, 100 and 500 kDa at concentration from 2 to 5 g/dL were used to suspend the RBCs. The radius and velocity of sedimenting RBC aggregates were investigated using image analysis. The radius and sedimentation velocity of aggregates increased initially, then decreased after achieving maxima. The maximal velocity of RBC aggregates showed a bell-shaped dependence on dextran molecular mass and concentration, whereas maximal radius showed monotonic increase with both factors. Difference between aggregate and solution density was estimated using aggregate radius and sedimentation velocity and dextran solution viscosity, and was consistent across most molecular mass and concentration levels. This allowed to calculate the porosity of aggregates and to show that it monotonically decreased with the increase in the solution density, caused by the increase in the dextran concentration. The results provide insight into the RBC aggregation process in solutions of proteins of different size, reflecting various pathological conditions. The currently reported data can be potentially applied to specific pathophysiological conditions giving an interpretation that is not yet fully discussed in the literature.

Erythrocyte aggregation in sudden flow arrest is linked to hyperthermia, hypoxemia, and band 3 availability.

BACKGROUND: Erythrocyte aggregation is a phenomenon that is commonly found in several pathological disease states: stroke, myocardial infarction, thermal burn injury, and COVID-19. Erythrocyte aggregation is characterized by rouleaux, closely packed stacks of cells, forming three-dimensional structures. Healthy blood flow monodisperses the red blood cells (RBCs) throughout the vasculature; however, in select pathological conditions, involving hyperthermia and hypoxemia, rouleaux formation remains and results in occlusion of microvessels with decreased perfusion. OBJECTIVES: Our objective is to address the kinetics of rouleaux formation with sudden cessation of flow in variable temperature and oxygen conditions. METHODS: RBCs used in this in vitro system were obtained from healthy human donors. Using a vertical stop-flow system aligned with a microscope, images were acquired and analyzed for increased variation in grayscale to indicate increased aggregation. The onset of aggregation after sudden cessation of flow was determined at proscribed temperatures (37-49°C) and oxygen (0%, 10%), and in the presence and absence of 4, 4'-Diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS). Both autologous and homologous plasma were tested. RESULTS: RBCs in autologous plasma aggregate faster and with a higher magnitude with both hyperthermia and hypoxemia. Preventing deoxyhemoglobin from binding to band 3 with DIDS (dissociates the cytoskeleton from the membrane) fully blocks aggregation. Further, RBC aggregation magnitude is greater in autologous plasma. CONCLUSIONS: We show that the C-terminal domain of band 3 plays a pivotal role in RBC aggregation. Further, aggregation is enhanced by hyperthermia and hypoxemia.

Red Blood Cell Morphodynamics in Patients with Polycythemia Vera and Stroke.

Polycythemia vera (PV) is a Ph-negative myeloproliferative neoplasm (MPN) which is characterized by erythrocytosis and a high incidence of thrombotic complications, including stroke. The study aimed to evaluate red blood cell (RBC) morphodynamic properties in PV patients and their possible association with stroke. We enrolled 48 patients with PV in this cross-sectional study, 13 of which have a history of ischemic stroke. The control group consisted of 90 healthy subjects. RBC deformability and aggregation analysis were performed using a laser-assisted optical rotational red cell analyzer. The following parameters were calculated: aggregation amplitude (Amp), RBC rouleaux formation time constant (Tf), time of formation of three-dimensional aggregates (Ts), aggregation index (AI), rate of complete disaggregation (y-dis), and the maximal elongation of RBC (EImax). Statistical analysis was performed with the R programming language. There were significant differences in RBCs morphodynamics features between patients with PV and the control group. Lower EImax (0.47 (0.44; 0.51) vs. 0.51 (0.47; 0.54), p < 0.001) and γ-dis (100 (100; 140) vs. 140 (106; 188) s-1, p < 0.001) along with higher amplitude (10.1 (8.6; 12.2) vs. 7.7 (6.6; 9.2), p < 0.001) was seen in patients with PV compared with control. A statistically significant difference between PV patients with and without stroke in aggregation amplitude was found (p = 0.03). A logistic regression model for stroke was built based on RBC morphodynamics which performed reasonably well (p = 0.01). RBC alterations may be associated with overt cerebrovascular disease in PV, suggesting a possible link between erythrocyte morphodynamics and increased risk of stroke.

Biosensing of Haemorheological Properties Using Microblood Flow Manipulation and Quantification.

The biomechanical properties of blood have been used to detect haematological diseases and disorders. The simultaneous measurement of multiple haemorheological properties has been considered an important aspect for separating the individual contributions of red blood cells (RBCs) and plasma. In this study, three haemorheological properties (viscosity, time constant, and RBC aggregation) were obtained by analysing blood flow, which was set to a square-wave profile (steady and transient flow). Based on a simplified differential equation derived using a discrete circuit model, the time constant for viscoelasticity was obtained by solving the governing equation rather than using the curve-fitting technique. The time constant (λ) varies linearly with respect to the interface in the coflowing channel (ß). Two parameters (i.e., average value: <λ>, linear slope: dλdß) were newly suggested to effectively represent linearly varying time constant. <λ> exhibited more consistent results than dλdß. To detect variations in the haematocrit in blood, we observed that the blood viscosity (i.e., steady flow) is better than the time constant (i.e., transient flow). The blood viscosity and time constant exhibited significant differences for the hardened RBCs. The present method was then successfully employed to detect continuously varying haematocrit resulting from RBC sedimentation in a driving syringe. The present method can consistently detect variations in blood in terms of the three haemorheological properties.

Blood rheometer based on microflow manipulation of continuous blood flows using push-and-back mechanism.

To understand the contributions of rheological properties to microcirculation, the simultaneous measurement of multiple rheological properties under continuous blood flows has been emphasized. However, existing methods exhibit limitations in terms of continuous and simultaneous monitoring. In this study, a simple method is suggested for simultaneously measuring four rheological properties (i.e., red blood cell (RBC) aggregation, blood viscosity, blood junction pressure, and RBC sedimentation) under a continuous blood flow. Using the push-and-back mechanism, which comprises a co-flowing channel, a test chamber, and an air compliance unit (ACU), blood is supplied to the test chamber and restored into the co-flowing channel periodically and reversely. First, RBC aggregation is quantified based on the intensity of the blood image in the test chamber. Second, blood viscosity and blood junction pressure are determined by analyzing the interface in the co-flowing channel. Lastly, RBC sedimentation is evaluated by analyzing the intensity of the blood image in the blood chamber. Based on quantitative studies involving several vital factors, the tubing length of ACU is set to L = 30 mm. The reference fluid (glycerin [20%]) is controlled in a periodic on-off manner (period = 240 s, and flow rate = 1 mL h-1). The blood flow rate is maintained at 1 mL h-1. Subsequently, the present method is used to determine the rheological properties of several blood samples with different hematocrits or diluents. Compared with previous studies, the present method yields sufficiently consistent trends with respect to the hematocrit level or concentration of dextran solution. The experimental results imply that the present method enables simultaneous and consistent measurements of four rheological properties of blood under continuous blood flows. This method can be regarded as a promising method for monitoring multiple rheological properties of blood circulating under an in vitro closed fluidic circuit.

Evaluation method of the degree of red blood cell aggregation considering ultrasonic propagation attenuation by analyzing ultrasonic backscattering properties.

PURPOSE: Red blood cell (RBC) aggregation is one of the main factors that determines blood viscosity and an important indicator for evaluating blood properties. As a noninvasive and quantitative method for diagnosing blood properties, our research group estimated the size of RBC aggregates by fitting the scattered power spectrum from the blood vessel lumen with the theoretical scattering characteristics to evaluate the degree of RBC aggregation. However, it was assumed that the propagation attenuation of ultrasound in the vascular lumen was the same regardless of whether RBCs were aggregated or not, which caused systematic errors in the estimated size. METHODS: To improve the size estimation accuracy, we calculated and corrected the attenuation of the blood vessel lumen during RBC aggregation and non-aggregation. The attenuation in the blood vessel lumen was calculated with the spectra acquired from two different depths. RESULTS: In the basic experiments using microparticles, the estimation accuracy decreased as the concentration increased in the case of the conventional method, but the estimated size tended to approach the true size irrespective of the concentration, removing the propagation attenuation component with the proposed method. In the in vivo experiment on the human hand dorsal vein, the size was estimated to be larger during RBC aggregation and smaller during non-aggregation using the proposed method. CONCLUSION: These results suggest that the proposed method can provide precise size estimation by considering the propagation attenuation component regardless of differences in blood conditions such as RBC concentration and degree of aggregation.

ß-Dispersion of blood during sedimentation.

Aggregation of human red blood cells (RBC) is central to various pathological conditions from bacterial infections to cancer. When left at low shear conditions or at hemostasis, RBCs form aggregates, which resemble stacks of coins, known as 'rouleaux'. We experimentally examined the interfacial dielectric dispersion of aggregating RBCs. Hetastarch, an RBC aggregation agent, is used to mimic conditions leading to aggregation. Hetastrach concentration is incrementally increased in blood from healthy donors to measure the sensitivity of the technique. Time lapse electrical impedance measurements were conducted as red blood cells form rouleaux and sediment in a PDMS chamber. Theoretical modeling was used for obtaining complex permittivity of an effective single red blood cell aggregate at various concentrations of hetastarch. Time response of red blood cells' impedance was also studied to parametrize the time evolution of impedance data. Single aggregate permittivity at the onset of aggregation, evolution of interfacial dispersion parameters, and sedimentation kinetics allowed us to distinguish differential aggregation in blood.

Ultrasound Standing Wave-Based Cell-to-liquid Separation for Measuring Viscosity and Aggregation of Blood Sample.

When quantifying mechanical properties of blood samples flowing in closed fluidic circuits, blood samples are collected at specific intervals. Centrifugal separation is considered as a required procedure for preparing blood samples. However, the use of centrifuge is associated with several issues, including the potential for red blood cell (RBC) lysis, clotting activation, and RBC adhesions in the tube. In this study, an ultrasonic transducer is employed to separate RBCs or diluent from blood sample. The ultrasonic radiation force is much smaller than the centrifugal force acting in centrifuge, it can avoid critical issues occurring under centrifuge. Then, the RBC aggregation and blood viscosity of the blood sample are obtained using the microfluidic technique. According to the numerical results, ultrasonic transducers exhibited a maximum quality factor at an excitation frequency of 2.1 MHz. Periodic pattern of acoustic pressure fields were visualized experimentally as a column mode. The half wavelength obtained was as 0.5 λ = 0.378 ± 0.07 mm. The experimental results agreed with the analytical estimation sufficiently. An acoustic power of 2 W was selected carefully for separating RBCs or diluent from various blood samples (i.e., Hct = 20% ~ 50%; diluent: plasma, 1x phosphate-buffered saline (PBS), and dextran solution). The present method was employed to separate fixed blood samples which tended to stack inside the tube while using the centrifuge. Fixed RBCs were collected easily with an ultrasonic transducer. After various fixed blood samples with different base solutions (i.e., glutaraldehyde solution, 1x PBS, and dextran solution) were prepared using the present method, RBC aggregation and the viscosity of the blood sample are successfully obtained. In the near future, the present method will be integrated into ex vivo or in vitro fluidic circuit for measuring multiple mechanical properties of blood samples for a certain longer period.

The impact of circulation in a heart-lung machine on function and survival characteristics of red blood cells.

Extracorporeal circulation is accompanied by changes in red blood cell morphology and structural integrity that affect cell function and survival, and thereby may contribute to the various side effects of heart-lung machine-assisted surgery. Our main objectives were to determine the effect of circulation of red blood cells in a stand-alone extracorporeal circuit on several parameters that are known to be affected by, as well as contribute to red blood cell aging. As a source of RBCs, we employed blood bank storage units of different ages. In order to assess the relevance of our in vitro observations for the characterization of extracorporal circulation technology, we compared these changes in those of patients undergoing extracorporeal circulation-assisted cardiac surgery. Our results show that circulation in a heart-lung machine is accompanied by changes in red blood cell volume, an increase in osmotic fragility, changes in deformability and aggregation behavior, and alterations in the exposure of phosphatidylserine and in microvesicle generation. RBCs from 1-week-old concentrates showed the highest similarities with the in vivo situation. These changes in key characteristics of the red blood cell aging process likely increase the susceptibility of red blood cells to the various mechanical, osmotic, and immunological stress conditions encountered during and after surgery in the patient's circulation, and thereby contribute to the side effects of surgery. Thus, aging-related parameters in red blood cell structure and function provide a foundation for the validation and improvement of extracorporeal circulation technology.

High-intensity interval training irrespective of its intensity improves markers of blood fluidity in hypertensive patients.

Aim: The present study examined and compared the effects of two different HIIT (High-intensity interval training) protocols on markers of blood fluidity in hypertensive patients.Methods: Thirty hypertensive (stage 1, systolic BP >140 and diastolic BP>90 mmHg) patients (age, 47.96 ± 3.20 yrs), were randomly allocated to short-duration HIIT (SDHIIT, n = 10), long-duration HIIT (LDHIIT, n = 10), and control (n = 10) groups. After 2 weeks of continuous mild training, patients in SDHIIT group performed 8 weeks of HIIT included 27 min HIIT that encompassed 27 repetitions of 30 s activity at 80%-100% of VO2peak interspersed by 30 s passive/active (10%-20% of VO2peak) recovery, while, patients in LDHIIT group performed 8 weeks of HIIT (32 min per session) included 4 repetitions of 4 min activity at 75%-90% of VO2peak interspersed by 4 min passive/active (15%-30% of VO2peak) recovery. Two blood samples were taken before and after training and were analyzed for hemorheological variables.Results: Significant (P < .05) reductions in systolic blood pressure (SBP), blood and plasma viscosity, fibrinogen concentration and red blood cell (RBC) aggregation (8-12%) were found following two training protocols (P < .05), though, the differences between adaptations were not statistically significant (P > .05). In addition, HIIT protocols increased RBC deformability significantly (P < .05), with no significant differences being observed between two protocols.Conclusion: It is concluded that HIIT training reduces SBP and markers of blood fluidity in patients with stage 1 hypertension irrespective of the HIIT intensity and duration. Therefore, this type of exercise training could be prescribed for improving the blood fluidity markers in hypertensive patients.

Pilot clinical study of quantitative ultrasound spectroscopy measurements of erythrocyte aggregation within superficial veins.

BACKGROUND: An enhanced inflammatory response is a trigger to the production of blood macromolecules involved in abnormally high levels of erythrocyte aggregation. OBJECTIVE: This study aimed at demonstrating for the first time the clinical feasibility of a non-invasive ultrasound-based erythrocyte aggregation quantitative measurement method for potential application in critical care medicine. METHODS: Erythrocyte aggregation was evaluated using modeling of the backscatter coefficient with the Structure Factor Size and Attenuation Estimator (SFSAE). SFSAE spectral parameters W (packing factor) and D (mean aggregate diameter) were measured within the antebrachial vein of the forearm and tibial vein of the leg in 50 healthy participants at natural flow and reduced flow controlled by a pressurized bracelet. Blood samples were also collected to measure erythrocyte aggregation ex vivo with an erythroaggregometer (parameter S10). RESULTS: W and Din vivo measurements were positively correlated with the ex vivoS10 index for both measurement sites and shear rates (correlations between 0.35-0.81, p < 0.05). Measurement at low shear rate was found to increase the sensitivity and reliability of this non-invasive measurement method. CONCLUSIONS: We behold that the SFSAE method presents systemic measures of the erythrocyte aggregation level, since results on upper and lower limbs were highly correlated.

Possible Error in Reflection Pulse Oximeter Readings as a Result of Applied Pressure.

Pulse oximetry is one of the most widely used techniques in modern medicine. In pulse oximetry, photoplethysmography (PPG) signals are measured at two different wavelengths and converted into the parameter Gamma, which is used to calculate the oxygen saturation of arterial blood. Although most pulse oximetry sensors are based on transmission geometry, the reflection mode is required for different form factors such as the forehead or wrists. In reflection oximetry, local pressure is applied to the measurement surface. We investigated the relationship between applied pressure and Gamma and found that for the reflection mode, Gamma tends to increase with increasing applied pressure. To explain this, we described the PPG signal in terms of two alternative models: a volumetric model and a Scattering-Driven Model (SDM). We assumed that the application of external pressure results in a decrease in local blood flow. We showed that only SDM correctly qualitatively describes Gamma as a function of the decrease in blood flow. We concluded that both described models coexist and that the relative influence of each depends on the measurement geometry and blood perfusion in the skin.

MONITORING OF HEMORHEOLOGICAL PARAMETERS IN PATIENTS WITH ATRIAL FIBRILLATION (INITIAL DATA).

The goals of this project were to determine how hemodynamics and hemorheological change in patients diagnosed with different forms of atrial fibrillation; also, how relievers of changes in fibrinogen monitoring in patients with permanent, persistent, paroxysmal forms of arrhythmias. There was examined 30 patients (the average age of patients 65). Patients and control groups have been tested the following studies: index of erythrocytes aggregability, deformability, plasma viscosity to evaluate the blood rheological parameter, also -Fibrinogen to determine of coagulation condition. The Index of the Erythrocytes aggregation was done with the system of textural analyzes. These new innovative methods "Georgian Technique" is created by Georgian scientists and they are famous in the world as direct, numeral and exact. The index of the deformation of the erythrocytes was done with filtration method. Plasma viscosity was measured in the capillary viscosimeter in 370 C. According to the obtained data patients with atrial fibrillation have the same conditions of rheological and coagulation systems, despite of the forms of the atrial fibrillation. And it is different compared to the control group. In the statistical processing of the total row of fibrinogen, the patients were divided into 3 categories. As it turned out fibrinogen and Index of erythrocytes aggregation are in a linear relationship. The quantity of fibrinogen and of erythrocytes aggregation increase simultaneously and the greater the sequential number of the category is, the changes are more pronounced. However, the change/variability of each biological parameter, as shown from our data, is not uniform and linear. The obtained results clearly illustrate the existence of two parallel mechanisms in the body. These are on the one hand the systems of coagulation, anti-coagulation and fibtinollysis. These processes are in a state of the weighted condition, and they are characterized by dynamic equilibrium. On the other hand, the rheology system, which involves the combination of blood flow, blood velocity, vascular stiffness / elasticity, is characterized by one direction going on, with the adequacy and non-existing antipodal mechanisms. The hemorheological system does not have a physiologically balanced opposing anti-rheological system. All of this generate that the hemorheological status is very important in the development and formation of some disease. The arrangement of a rheologycal system is taking preventive character. On the one hand, the rheologycal system is a consolidation of diagnosis and evaluation of the mechanisms. Also, it is the treatment target. Normalization of them is very important in the therapeutic standpoint of the individual.

The effects of stenting on hemorheological parameters: An in vitro investigation under various blood flow conditions.

 Despite their wide clinical usage, stent functionality may be compromised by complications at the site of implantation, including early/late stent thrombosis and occlusion. Although several studies have described the effect of fluid-structure interaction on local haemodynamics, there is yet limited information on the effect of the stent presence on specific hemorheological parameters. The current work investigates the red blood cell (RBC) mechanical behavior and physiological changes as a result of flow through stented vessels. Blood samples from healthy volunteers were prepared as RBC suspensions in plasma and in phosphate buffer saline at 45% haematocrit. Self-expanding nitinol stents were inserted in clear perfluoroalkoxy alkane tubing which was connected to a syringe, and integrated in a syringe pump. The samples were tested at flow rates of 17.5, 35 and 70 ml/min, and control tests were performed in non-stented vessels. For each flow rate, the sample viscosity, RBC aggregation and deformability, and RBC lysis were estimated. The results indicate that the presence of a stent in a vessel has an influence on the hemorheological characteristics of blood. The viscosity of all samples increases slightly with the increase of the flow rate and exposure. RBC aggregation and elongation index (EI) decrease as the flow rate and exposure increases. RBC lysis for the extreme cases is evident. The results indicate that the stresses developed in the stent area for the extreme conditions could be sufficiently high to influence the integrity of the RBC membrane.

Quantitative Measurement and Evaluation of Red Blood Cell Aggregation in Normal Blood Based on a Modified Hanai Equation.

The aggregation of red blood cells (RBCs) in normal blood (non-coagulation) has been quantitatively measured by blood pulsatile flow based on multiple-frequency electrical impedance spectroscopy. The relaxation frequencies fc under static and flowing conditions of blood pulsatile flow are utilized to evaluate the RBC aggregation quantitatively with the consideration of blood flow factors (RBC orientation, deformation, thickness of electrical double layer (EDL)). Both porcine blood and bovine blood are investigated in experiments, for the reason that porcine blood easily forms RBC aggregates, while bovine blood does not. The results show that the relaxation frequencies fc of porcine blood and bovine blood present opposite performance, which indicates that the proposed relaxation frequency fc is efficient to measure RBCs aggregation. Furthermore, the modified Hanai equation is proposed to quantitatively calculate the influence of RBCs aggregation on relaxation frequency fc. The study confirms the feasibility of a high speed, on-line RBC aggregation sensing method in extracorporeal circulation systems.

Red blood cell aggregation, disaggregation and aggregate morphology in autologous plasma and serum in diabetic foot disease.

BACKGROUND: Diabetes mellitus is frequently associated with microcirculation pathology and hemorheological disorders. METHODS: 24 patients with diabetic foot and 22 healthy subjects were recruited. RBC aggregation, disaggregation and morphology of aggregates were determined in autologous plasma and serum. RESULTS: The RBC aggregation in patients with diabetic foot increased in autologous plasma and serum. Increased red blood cell aggregate strength in these patients was observed only in autologous plasma. Microscopic images of RBC aggregates of patients with diabetic foot show the formation of pathologic globular structures of aggregates in autologous plasma and serum. CONCLUSION: The RBC aggregation in autologous plasma and autologous serum in patients with diabetic foot is significantly higher than in healthy subjects. Increase in strength of RBC aggregates in diabetic foot patients was observed only in autologous plasma. The microscopic images of RBC aggregates in patients with diabetic foot indicate the formation of globular (pathologic) structures of aggregates in autologous plasma and serum. The differences in the morphology of RBC aggregates in autologous plasma and serum between healthy subjects and diabetic foot patients, obtained by microscopic image analysis with high magnification light microscope, can be used as an additional diagnostic tool in medical practice.

Fibrinogen-erythrocyte binding and hemorheology measurements in the assessment of essential arterial hypertension patients.

Some studies have reported a positive association between plasma fibrinogen levels, erythrocyte aggregation and essential arterial hypertension (EAH). The aim of this study was to understand how the interaction between fibrinogen and its erythrocyte membrane receptor is altered in EAH. EAH patients (n = 31) and healthy blood donors (n = 65) were enrolled in the study. EAH patients were therapeutically controlled for the disease, presenting a systolic blood pressure between 108 and 180 mmHg and a diastolic blood pressure between 66 and 123 mmHg. Clinical evaluation included blood pressure monitoring, electrocardiography, echocardiography and blood cell count. The hemorheological parameters were also analyzed. Fibrinogen-erythrocyte binding force and frequency were evaluated quantitatively, at the single-molecule level, using atomic force microscopy (AFM). Changes in erythrocyte elasticity were also evaluated. Force spectroscopy data showed that the average fibrinogen-erythrocyte binding forces increase from 40.4 ± 3.0 pN in healthy donors to 73.8 ± 8.1 pN in patients with EAH, despite a lower binding frequency for patients compared to the control group (7.9 ± 1.6% vs. 27.6 ± 4.2%, respectively). Elasticity studies revealed an increase of erythrocyte stiffness in the patients. The stronger fibrinogen binding to erythrocytes from EAH patients and alteration in cell elasticity may lead to changes in the whole blood flow. The patients' altered hemorheological parameters may also contribute to these blood flow perturbations. The transient bridging of two erythrocytes, by the simultaneous binding of fibrinogen to both of them, promoting erythrocyte aggregation, could represent an important cardiovascular risk factor.