Shear-thinning behaviour of blood in response to active hyperaemia: Implications for the assessment of arterial shear stress-mediated dilatation.

NEW FINDINGS: What is the central question of this study? Quantitative values of shear rate-specific blood viscosity and shear stress in the human macrovasculature in response to exercise hyperaemia are unknown. What is the main finding and its importance? Using the handgrip exercise model, we showed that an increase in brachial artery shear rate led to a decrease in blood viscosity, despite concomitant haemoconcentration. This shear-thinning behaviour of blood, secondary to increased erythrocyte deformability, blunted the expected increase in brachial artery shear stress based on shear rate prediction. Our data yield new insights into the magnitude and regulation of macrovascular blood viscosity and shear stress in physiological conditions of elevated metabolic demand and blood flow in humans. ABSTRACT: Blood viscosity is a well-known determinant of shear stress and vascular resistance; however, accurate quantitative assessments of shear rate-specific blood viscosity in the macrovasculature in conditions of elevated blood flow are inherently difficult, owing to the shear-thinning behaviour of blood. Herein, 12 men performed graded rhythmic handgrip exercise at 20, 40, 60 and 80% of their maximal workload. Brachial artery shear rate and diameter were measured via high-resolution Duplex ultrasound. Blood was sampled serially from an i.v. cannula in the exercising arm for the assessment of blood viscosity (cone-plates viscometer). We measured ex vivo blood viscosity at 10 discrete shear rates within the physiological range documented for the brachial artery in basal and exercise conditions. Subsequently, the blood viscosity data were fitted with a two-phase exponential decay, facilitating interpolation of blood viscosity values corresponding to the ultrasound-derived shear rate. Brachial artery shear rate and shear stress increased in a stepwise manner with increasing exercise intensity, reaching peak values of 940 ± 245 s-1 and 3.68 ± 0.92 Pa, respectively. Conversely, brachial artery shear rate-specific blood viscosity decreased with respect to baseline values throughout all exercise intensities by ∼6-11%, reaching a minimal value of 3.92 ± 0.35 mPa s, despite concomitant haemoconcentration. This shear-thinning behaviour of blood, secondary to increased erythrocyte deformability, blunted the expected increase in shear stress based on shear rate prediction. Consequently, the use of shear stress yielded a higher slope for the brachial artery stimulus versus dilatation relationship than shear rate. Collectively, our data refute the use of shear rate to infer arterial shear stress-mediated processes.

Optofluidic laser speckle image decorrelation analysis for the assessment of red blood cell storage.

Red blood cells (RBCs) undergo irreversible biochemical and morphological changes during storage, contributing to the hemorheological changes of stored RBCs, which causes deterioration of microvascular perfusion in vivo. In this study, a home-built optofluidic system for laser speckle imaging of flowing stored RBCs through a transparent microfluidic channel was employed. The speckle decorrelation time (SDT) provides a quantitative measure of RBC changes, including aggregation in the microchannel. The SDT and relative light transmission intensity of the stored RBCs were monitored for 42 days. In addition, correlations between the decorrelation time, RBC flow speed through the channel, and relative light transmission intensity were obtained. The SDT of stored RBCs increased as the storage duration increased. The SDTs of the RBCs stored for 21 days did not significantly change. However, for the RBCs stored for over 35 days, the SDT increased significantly from 1.26 ± 0.27 ms to 6.12 ± 1.98 ms. In addition, we measured the relative light transmission intensity and RBC flow speed. As the RBC storage time increased, the relative light transmission intensity increased, whereas the RBC flow speed decreased in the microchannel. The optofluidic laser speckle image decorrelation time provides a quantitative measure of assessing the RBC condition during storage. Laser speckle image decorrelation analysis may serve as a convenient assay to monitor the property changes of stored RBCs.

Viscosity of Plasma as a Key Factor in Assessment of Extracellular Vesicles by Light Scattering.

Extracellular vesicles (EVs) isolated from biological samples are a promising material for use in medicine and technology. However, the assessment methods that would yield repeatable concentrations, sizes and compositions of the harvested material are missing. A plausible model for the description of EV isolates has not been developed. Furthermore, the identity and genesis of EVs are still obscure and the relevant parameters have not yet been identified. The purpose of this work is to better understand the mechanisms taking place during harvesting of EVs, in particular the role of viscosity of EV suspension. The EVs were harvested from blood plasma by repeated centrifugation and washing of samples. Their size and shape were assessed by using a combination of static and dynamic light scattering. The average shape parameter of the assessed particles was found to be ρ ~ 1 (0.94-1.1 in exosome standards and 0.7-1.2 in blood plasma and EV isolates), pertaining to spherical shells (spherical vesicles). This study has estimated the value of the viscosity coefficient of the medium in blood plasma to be 1.2 mPa/s. It can be concluded that light scattering could be a plausible method for the assessment of EVs upon considering that EVs are a dynamic material with a transient identity.

Testosterone Therapy: An Assessment of the Clinical Consequences of Changes in Hematocrit and Blood Flow Characteristics.

INTRODUCTION: Clinical guidelines indicate that hematocrit should be monitored during testosterone replacement therapy (TTh), with action taken if a level of 0.54 is exceeded. AIM: To consider the extent of changes in hematocrit and putative effects on viscosity, blood flow, and mortality rates after TTh. METHODS: We focused on literature describing benefits and possible pitfalls of TTh, including increased hematocrit. We used data from the BLAST RCT to determine change in hematocrit after 30 weeks of TTh and describe a clinical case showing the need for monitoring. We consider the validity of the current hematocrit cutoff value at which TTh may be modified. Ways in which hematocrit alters blood flow in the micro- and macro-vasculature are also considered. MAIN OUTCOME MEASURES: The following measures were assessed: (i) change in hematocrit, (ii) corresponding actions taken in clinical practice, and (iii) possible blood flow changes following change in hematocrit. RESULTS: Analysis of data from the BLAST RCT showed a significant increase in mean hematocrit of 0.01, the increase greater in men with lower baseline values. Although 0 of 61 men given TTh breached the suggested cutoff of 0.54 after 30 weeks, a clinical case demonstrates the need to monitor hematocrit. An association between hematocrit and morbidity and mortality appears likely but not proven and may be evident only in patient subgroups. The consequences of an increased hematocrit may be mediated by alterations in blood viscosity, oxygen delivery, and flow. Their relative impact may vary in different vascular beds. CONCLUSIONS: TTh can effect an increased hematocrit via poorly understood mechanisms and may have harmful effects on blood flow that differ in patient subgroups. At present, there appears no scientific basis for using a hematocrit of 0.54 to modify TTh; other values may be more appropriate in particular patient groups. König CS, Balabani S, Hackett GI, et al. Testosterone Therapy: An Assessment of the Clinical Consequences of Changes in Hematocrit and Blood Flow Characteristics. Sex Med Rev 2019;7:650-660.

Assessment of viscous energy loss and the association with three-dimensional vortex ring formation in left ventricular inflow: In vivo evaluation using four-dimensional flow MRI.

PURPOSE: To evaluate viscous energy loss and the association with three-dimensional (3D) vortex ring formation in left ventricular (LV) blood flow during diastolic filling. THEORY AND METHODS: Thirty healthy volunteers were compared with 32 patients with corrected atrioventricular septal defect as unnatural mitral valve morphology and inflow are common in these patients. 4DFlow MRI was acquired from which 3D vortex ring formation was identified in LV blood flow at peak early (E)-filling and late (A)-filling and characterized by its presence/absence, orientation, and position from the lateral wall. Viscous energy loss was computed over E-filling, A-filling, and complete diastole using the Navier-Stokes energy equations. RESULTS: Compared with healthy volunteers, viscous energy loss was significantly elevated in patients with disturbed vortex ring formation as characterized by a significantly inclined orientation and/or position closer to the lateral wall. Highest viscous energy loss was found in patients without a ring-shaped vortex during E-filling (on average more than double compared with patients with ring-shape vortex, P < 0.003). Altered A-filling vortex ring formation was associated with significant increase in total viscous energy loss over diastole even in the presence of normal E-filling vortex ring. CONCLUSION: Altered vortex ring formation during LV filling is associated with increased viscous energy loss. Magn Reson Med 77:794-805, 2017. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

IgMκ and IgMλ Measurements for the Assessment of Patients with Waldenström's Macroglobulinaemia.

PURPOSE: Accurate quantification of monoclonal IgM immunoglobulins is essential for response assessment in patients with Waldenström's macroglobulinaemia (WM). The propensity of IgM to form multimers in serum makes sample evaluation by current laboratory methods particularly challenging. EXPERIMENTAL DESIGN: We assessed the precision and linearity of IgMκ and IgMλ heavy/light chain (HLC, Hevylite) assays, and established reference intervals using 120 normal donor sera. We compared the quantitative performance of HLC assays with serum protein electrophoresis (SPE) and total IgM nephelometry for 78 diagnostic samples and follow-up samples from 25 patients with WM. Comparisons were made between the three methods for diagnostic sensitivity and response assessment. RESULTS: IgMκ and IgMλ HLC assays showed low imprecision and good linearity. There was good agreement between summated HLC (IgMκ + IgMλ) and total IgM (measured nephelometrically; R2 = 0.90), but only moderate agreement between involved IgM HLC and SPE densitometry (R2 = 0.49). Analysis of 120 normal donor sera produced the following normal ranges: IgMκ: 0.29-1.82 g/L; IgMλ: 0.17-0.94 g/L; IgMκ/IgMλ ratio: 0.96-2.30. Using these ranges, IgM HLC ratios were abnormal in all WM presentation sera tested, including 15 with non-quantifiable SPE. Despite discordance in quantitation, responses assigned with HLC assays showed excellent agreement to those based on international guidelines using SPE or total IgM; although abnormal HLC ratios indicated residual disease in some patients with negative electrophoresis results. CONCLUSIONS: Nephelometric assessment of IgMκ and IgMλ HLC pairs offers a quantitative alternative to traditional laboratory techniques for the measurement of monoclonal IgM and may aid in the management of WM. Clin Cancer Res; 22(20); 5152-8. ©2016 AACR.

Contribution of blood viscosity in the assessment of flow-mediated dilation and arterial stiffness.

Flow-mediated dilation (FMD) is a non-invasive index of endothelial function. In an attempt to standardize FMD for shear stimulus, shear rate (velocity/diameter), rather than shear stress (viscosity*velocity/diameter), is commonly used as a surrogate measure, although it is limited by individual differences in blood viscosity. The purpose of this study was to determine the contribution of whole blood viscosity to FMD and other key measures of vascular function. Blood viscosity, FMD, carotid artery compliance, and carotid-femoral pulse wave velocity (cfPWV) were measured in 98 apparently healthy adults varying widely in age (18-63 years). Whole blood viscosity was not significantly correlated with FMD, cfPWV, or carotid artery compliance. Shear rate was a stronger correlate with FMD than shear stress that takes blood viscosity into account (r = 0.43 vs 0.28). No significant differences were observed between whole blood viscosity and traditional risk factors for cardiovascular disease. Age was positively correlated with cfPWV (r = 0.65, p < 0.001) and negatively correlated with FMD (r = -0.24, p < 0.05) and carotid artery compliance (r = -0.45, p < 0.01). Controlling for viscosity did not reduce the strength of these relations. These results indicate that whole blood viscosity does not significantly impact measures of vascular function and suggests that the common practice to use shear rate, rather than shear stress, in the adjustment of FMD is valid.

Cardiac mechanoenergetic cost of elevated plasma viscosity after moderate hemodilution.

The purpose of this study was to investigate how plasma viscosity affects cardiac and vascular function during moderate hemodilution. Twelve anesthetized hamsters were hemodiluted by 40% of blood volume with two different viscosity plasma expanders. Experimental groups were based on the plasma expander viscosity, namely: high viscosity plasma expander (HVPE, 6.3 mPa · s) and low viscosity plasma expander (LVPE, 2.2 mPa · s). Left ventricular (LV) function was intracardiacally measured with a high temporal resolution miniaturized conductance catheter and concurrent pressure-volume results were used to calculate different LV indices. Independently of the plasma expander, hemodilution decreased hematocrit to 28% in both groups. LVPE hemodilution reduced whole blood viscosity by 40% without changing plasma viscosity, while HVPE hemodilution reduced whole blood viscosity by 23% and almost doubled plasma viscosity relative to baseline. High viscosity plasma expander hemodilution significantly increased cardiac output, stroke volume and stroke work compared to baseline, whereas LVPE hemodilution did not. Furthermore, an increase in plasma viscosity during moderate hemodilution produced a higher energy transfer per unit volume of ejected blood. Systemic vascular resistance decreased after hemodilution in both groups. Counter-intuitively, HVPE hemodilution showed lower vascular resistance and vascular hindrance than LVPE hemodilution. This result suggests that geometrical changes in the circulatory system are induced by the increase in plasma viscosity. In conclusion, an increase in plasma viscosity after moderate hemodilution directly influenced cardiac and vascular function by maintaining hydraulic power and reducing systemic vascular resistance through vasodilation.

Assessment of blood coagulation under various flow conditions with ultrasound backscattering.

Several in vitro studies have employed ultrasonic techniques to detect varying properties of coagulating blood under static or stirred conditions. Most of those studies mainly addressed on the development and feasibility of modalities and however were not fully considering the effect of blood flow. To better elucidate this issue, ultrasonic backscattering were measured from the coagulating porcine blood circulated in a mock flow loop with various steady laminar flows at mean shear rates from 10 to 100 s(-1). A 3 ml of 0.5 M CaCl2 solution for inducing blood coagulation was added to that of 30 ml blood circulated in the conduit. For each measurement carried out with a 10-MHz transducer, backscattered signals digitized at 100-MHz sampling frequency were acquired for a total of 20 min at temporal resolution of 50 A-lines per s. The integrated backscatter (IB) was calculated for assessing backscattering properties of coagulating blood. The results show that blood coagulation tended to be increased corresponding to the addition of CaCl2 solution: the IB was increased approximately 6.1 +/- 0.6 (mean +/- standard deviation), 5.4 +/- 0.9, and 4.5 +/- 1.2 dB at 310 +/- 62, 420 +/- 88, and 610 +/- 102 s associated with mean shear rates of 10, 40, and 100 s(-1), respectively. The rate of increasing IB for evaluating the growth of clot was estimated to be 0.075 +/- 0.017, 0.052 +/- 0.027, and 0.038 +/- 0.012 delta dB delta s(-1) corresponding to the increase of mean shear rates. These results consistently demonstrate that higher shear rate tends to prolong the duration for the flowing blood to be coagulated and to decrease the rate of IB. Moreover, the laminar flow was changed to turbulent flow during that the blood was clotting discerned by spatial variations of ultrasound backscattering in the conduit. All these results validate that ultrasound backscattering is feasible to be utilized for detecting and assessing blood coagulation under dynamic conditions.

Noninvasive simultaneous assessment of wall shear rate and wall distension in carotid arteries.

A novel technique has been developed for the noninvasive real-time simultaneous assessment of both blood velocity profile and wall displacements in human arteries. The novel technique is based on the use of two ultrasound beams, one set at optimal angle for wall motion measurements and the other for blood velocity profile measurements. The technique was implemented on a linear array probe divided into two subapertures. A modified commercial ultrasound machine and a custom PC board based on a high-speed digital signal processor was used to process the quadrature demodulated echo signals and display results in realtime. Flow phantom experiments demonstrated the validity of the technique, providing wall shear rate (WSR) estimates within 10% of the theoretical values. The system was also tested in the common carotid arteries of 16 healthy volunteers (age 30 to 53 y). Results of simultaneous diameter distension and WSR measurements were in agreement with published data.

Assessment by transient elastography of the viscoelastic properties of blood during clotting.

Blood clotting is a natural process that can be both beneficial and life-threatening for the human body. It allows the maintenance of hemostasis after vascular injury, but it can also cause deep vein thrombosis and heart stroke. This study aimed better to understand the clotting process from a biomechanical point of view by using an acoustic method. The long-term objective is the staging of the age of clots in deep veins for therapy planning. The transient elastography method using a shear elasticity probe served to evaluate the shear wave velocity (V(S)) and shear wave attenuation (alpha(S)) of porcine whole blood during in vitro clot formation. By solving an inverse problem, it was then possible to provide images of the elasticity (mu(B)) and of the viscosity (eta(B)) from clotting blood. The time-varying elasticity and viscosity were very similar to what has been observed for the sol-gel transition of polymers. The mechanical properties of blood clot, which were modified by varying the hematocrit and by adding heparin or fibrinogen, were clearly assessed by the transient elastography technique. It is concluded that the shear elasticity probe is an appropriate tool to quantify and follow the sol-gel transition of blood during clotting.

Precision assessment of biofluid viscosity measurements using molecular rotors.

Blood viscosity changes with many pathologic conditions, but its importance has not been fully investigated because the current methods of measurement are poorly suited for clinical applications. The use of viscosity-sensitive fluorescent molecular rotors to determine fluid viscosity in a nonmechanical manner has been investigated recently, but it is unknown how the precision of the fluorescence-based method compares to established mechanical viscometry. Human blood plasma viscosity was modulated with high-viscosity plasma expanders, dextran, pentastarch, and hetastarch. The samples were divided into a calibration and a test set. The relationship between fluorescence emission and viscosity was established using the calibration set. Viscosity of the test set was determined by fluorescence and by cone-and-plate viscometer, and the precision of both methods compared. Molecular rotor fluorescence intensity showed a power law relationship with solution viscosity. Mechanical measurements deviated from the theoretical viscosity value by less than 7.6%, while fluorescence-based measurements deviated by less than 6%. The average coefficient of variation was 6.9% (mechanical measurement) and 3.4% to 3.8% (fluorescence-based measurement, depending on the molecular rotor used). Fluorescence-based viscometry exhibits comparable precision to mechanical viscometry. Fluorescence viscometry does not apply shear and is therefore more practical for biofluids which have apparent non-Newtonian properties. In addition, fluorescence instrumentation makes very fast serial measurements possible, thus promising new areas of application in laboratory and clinical settings.

Non-invasive assessment of systemic elastic behaviour in hypertensive patients: analysis of possible determinants.

Knowledge about the viscoelastic behaviour of the arterial wall has been proved to have physiological importance and clinical usage. Our purpose was to study the changes of the systemic arterial wall's elastic properties non-invasively, in patients with established essential and with borderline hypertension, and to evaluate its possible determinants. Three groups of normotensive, borderline and established essential hypertensive patients were evaluated. Arterial pulse wave velocity (PWV) was measured and arterial compliance (Cm) was derived in all patients. Pulse wave velocity was obtained from the pressure values of digitized carotid and radial arteries. Arterial compliance (Cm = dD/dP with P pressure and D diameter) was calculated using a formula derived from the Bramwell and Hill equation: Cm = (1,334 x D)/(2 rho x PWV2), where for D humeral diameter was used as measured by high resolution echograph, and rho is the blood density (rho = 1.06). Pulse wave velocity was significantly higher in established essential hypertensive patients with respect to normotensive patients (p < 0.05). Arterial compliance was significantly diminished in established and in borderline hypertensive patients with respect to normotensive patients (p < 0.05), which implies early alterations in hypertensive cardiovascular disease. Multiple regression analysis of the cofactors showed that age and diastolic pressure are independent determinants of Cm. Impairment of the arterial wall's intrinsic elastic properties was demonstrated in established essential hypertension, independent of age and diastolic pressure.

Flow instabilities induced by coronary artery stents: assessment with an in vitro pulse duplicator.

An in vitro pulse duplicator system was used to investigate whether coronary artery stents induce downstream flow instabilities. Hot film or electrochemical probes were used to measure wall shear stress before and after deployment of both single and multiple (overlapping) stents in normal and diseased coronary geometries. Left main coronary diameters ranged from 4 to 5 mm, whereas left anterior descending (LAD) and left circumflex (LCX) diameters ranged from 2 to 4 mm. Under resting conditions, all coronary flow waveforms remained laminar, even after stent placement. However, disturbances were found downstream from a stent placed in the proximal LAD under mild exercise conditions. These disturbances were found 5 mm distal to the stent, in both the LAD and the proximal LCX. Turbulence intensities of order 5% were induced by a single slotted stent in a normal LAD geometry. In cases of distal disease, the turbulence intensity was 9% with one stent and 11% with tandem stents. In cases of proximal disease, these values were 19 and 25%, respectively. The shear stress from these disturbances (20-200 dynes cm-2) is sufficient to delay re-endothelialization and promote restenosis. Therefore, the disturbances could contribute to the increased incidence of restenosis reported with multiple stents, and with stents used in cases of diffuse coronary disease.

Assessment of prosthetic valve hemodynamics by Doppler: lessons from in vitro studies of the St. Jude valve.

Although Doppler assessment of native heart valve hemodynamics is quite accurate, assessment of prosthetic mechanical valve hemodynamics presents more potential problems. These problems arise from the unique geometry of prosthetic valves. Two important prosthetic valve phenomena can violate the basic assumptions underlying Doppler calculations: pressure recovery and localized gradients. When pressure recovery is present, Doppler systematically overestimates the actual net catheter gradient across the valve. In contrast, the presence of localized gradients may cause Doppler to measure differing gradients depending on which part of the valve is interrogated. These phenomena can also affect the calculation of prosthetic valve areas using the continuity equation. When assessing prosthetic valves with Doppler echocardiography, it must be remembered that Doppler measures velocity, not pressure gradients and that the echocardiographer must exercise great care when interpreting Doppler velocities as either pressure gradients or in calculating valve areas for prosthetic valves.