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Background: Described to be antithrombotic and antihypertensive, nattokinase is consumed for putative cardiovascular benefit. However, no large-scale, long-term cardiovascular study has been conducted with nattokinase supplementation., Objective: To determine the effect of nattokinase on subclinical atherosclerosis progression and atherothrombotic biomarkers., Methods: In this double-blinded trial, 265 individuals of median age 65.3 years, without clinical evidence of cardiovascular disease (CVD) were randomized to oral nattokinase 2,000 fibrinolytic units or matching placebo. Primary outcome was rate of change in subclinical atherosclerosis measured by serial carotid ultrasound every 6 months as carotid artery intima-media thickness (CIMT) and carotid arterial stiffness (CAS). Additional outcomes determined at least every 6 months were clinical parameters including blood pressure and laboratory measures including metabolic factors, blood rheology parameters, blood coagulation and fibrinolysis factors, inflammatory markers and monocyte/macrophage cellular activation markers., Results: After median 3 years of randomized treatment, annualized rate of change in CIMT and CAS did not significantly differ between nattokinase supplementation and placebo. Additionally, there was no significant effect of nattokinase supplementation on blood pressure or any laboratory determination., Conclusions: Results of this trial show that nattokinase supplementation has a null effect on subclinical atherosclerosis progression in healthy individuals at low risk for CVD.

While a number of studies have shown short-term beneficial effects of therapeutic plasma exchange (TPE) for treating systemic scleroderma (SSc), there have been no reports on the very long-term usage of TPE as the sole systemic treatment intervention. We report the case of a male patient, originally diagnosed with limited systemic scleroderma (lcSSc) in early 1990, who has been undergoing regular plasmapheresis treatments for more than 22 years, beginning in late 1993. Prior to commencing treatment, the patient exhibited symptoms including severe gastro esophageal reflux disease (GERD) with esophagitis, frequent Raynaud's attacks, reduced lung function, and chronic chilling. With the exception of mild residual Raynaud's, all of the patient's symptoms reversed after three years of regular TPE treatments and he remains in complete remission. While the typical explanation for the therapeutic benefits seen with TPE focuses on temporary reduction of circulating antibodies or other pathogenic factors, we propose instead an explanation based on abnormal blood rheology as a novel disease pathogenesis model for SSc.

Background: Current generation mechanical circulatory assist devices are designed to minimize high shears to blood for prolonged durations to avoid hemolysis. However, red blood cells (RBC) demonstrate impaired capacity to deform when exposed to shear stress (SS) well below the "hemolytic threshold"., Objective: We endeavored to identify how changes in the magnitude and duration of SS exposure alter RBC deformability and subsequently develop a model to predict erythrocyte subhemolytic damage., Methods: RBC suspensions were exposed to discrete magnitudes of SS (1-64 Pa) for specific durations (1-64 s), immediately prior to RBC deformability being measured. Analyses included exploring the maximal RBC deformation (EImax) and SS required for half EImax (SS1/2). A surface-mesh was interpolated onto the raw data to predict impaired RBC deformability., Results: When SS was applied at <16Pa, limited changes were observed. When RBC were exposed to 32 Pa, mild impairments in EImax and SS1/2 occurred, although 64 Pa caused a dramatic impairment of RBC deformability. A clear relation between SS duration and magnitude was determined, which could predict impaired RBC deformability., Conclusion: The present results provide a model that may be used to predict whether RBC deformability is decreased following exposure to a given level and duration of SS, and may guide design of future generations of mechanical circulatory assist devices.

Background: Cell-cell and cell-surface adhesion modulated by water-soluble polymers continues to be of current interest, especially since prior reports have indicated a role for depletion-mediated attractive forces., Objective: To determine the effects of concentration and molecular mass of the neutral polymer dextran (40 kDa to 28 MDa) on the adhesion of human red blood cells (RBC) to coated glass coverslips., Methods: Confocal-reflection interference contrast microscopy (C-IRM), in conjunction with phase contrast imaging, was utilized to measure the adhesion dynamics and contact mechanics of RBC during the initial stages of cell contact with several types of substrates., Results: Adhesion is markedly increased in the presence of dextran with a molecular mass ⩾ 70 kDa. This increased adhesiveness is attributed to reduced surface concentration of the large polymers and hence increased attractive forces due to depletion interaction. The equilibrium deformation of adhering RBC was modeled as a truncated sphere and the calculated adhesion energies were in close agreement with theoretical results., Conclusions: These results clearly demonstrate that polymer depletion can promote RBC adhesion to artificial surfaces and suggest that this phenomenon may play a role in other specific and non-specific cell-cell interactions, such as rouleau formation and RBC-endothelial cell adhesion.

Sickle cell disease (SCD) is characterized by decreased erythrocyte deformability, microvessel occlusion and severe painful infarctions of different organs. Ektacytometry of SCD red blood cells (RBC) is made difficult by the presence of rigid, poorly-deformable irreversibly sickled cells (ISC) that do not align with the fluid shear field and distort the elliptical diffraction pattern seen with normal RBC. In operation, the computer software fits an outline to the diffraction pattern, then reports an elongation index (EI) at each shear stress based on the length and width of the fitted ellipse: EI=(length-width)/(length+width). Using a commercial ektacytometer (LORCA, Mechatronics Instruments, The Netherlands) we have approached the problem of ellipse fitting in two ways: (1) altering the height of the diffraction image on a computer monitor using an aperture within the camera lens; (2) altering the light intensity level (gray level) used by the software to fit the image to an elliptical shape. Neither of these methods affected deformability results (elongation index-shear stress relations) for normal RBC but did markedly affect results for SCD erythrocytes: (1) decreasing image height by 15% and 30% increased EI at moderate to high stresses; (2) progressively increasing the light level increased EI over a wide range of stresses. Fitting data obtained at different image heights using the Lineweaver-Burke routine yielded percentage ISC results in good agreement with microscopic cell counting. We suggest that these two relatively simple approaches allow minimizing artifacts due to the presence of rigid discs or ISC and also suggest the need for additional studies to evaluate the physiological relevance of deformability data obtained via these methods.

Consumption of red wine is associated with a decreased risk of several cardiovascular diseases (e.g., coronary artery disease, stroke), but unfortunately literature reports regarding ethanol's effects on hemorheological parameters are not concordant. In the present study, red blood cell (RBC) deformability was tested via laser ektacytometry (LORCA, 0.3-30 Pa) using two approaches: 1) addition of ethanol to whole blood at 0.25%-2% followed by incubation and testing in ethanol-free LORCA medium; 2) addition of ethanol to the LORCA medium at 0.25%-6% then testing untreated native RBC in these media. The effects of ethanol on deformability for oxidatively stressed RBC were investigated as were changes of RBC aggregation (Myrenne Aggregometer) for cells in autologous plasma or 3% 70 kDa dextran. Significant dose-related increases of RBC deformability were observed at 0.25% (p < 0.05) and higher concentrations only if ethanol was in the LORCA medium; no changes occurred for cells previously incubated with ethanol then tested in ethanol-free medium. The impaired deformability of cells pre-exposed to oxidative stress was improved only if ethanol was in the LORCA medium. RBC aggregation decreased with concentration at 0.25% and higher for cells in both autologous plasma and dextran 70. Our results indicate that ethanol reversibly improves erythrocyte deformability and irreversibly decreases erythrocyte aggregation; the relevance of these results to the health benefits of moderate wine consumption require further investigation.

Background: Previous studies have demonstrated that red blood cells (RBC) either lyse or at least experience mechanical damage following prolonged exposure to high shear stress (≥100 Pa). Conversely, prolonged shear stress exposure within the physiological range (5-20 Pa, 300 s) was recently reported to improve RBC deformability. This study investigated the relationships between shear stress and RBC deformability to determine the breakpoint between beneficial vs. detrimental exposure to shear stress (i.e., "subhemolytic threshold"). A second aim of the study was to determine whether the frequency of intermittent application of shear stress influenced the subhemolytic threshold., Methods: RBC were exposed to various levels of shear stress (0-100 Pa) in a Couette type shearing system for 300 s. RBC deformability was then immediately measured via ektacytometry. Parallel experiments were conducted at the same shear stresses, except the application time differed while keeping constant the total exposure time: shear stress was applied either for 30 s and repeated 10 times (10×30 s) or applied for 15 s and repeated 20 times (20×15 s)., Results: For a range of donors, the subhemolytic threshold with constant shear stress application was between 30-40 Pa. When physiological shear stress was applied in an intermittent manner, more frequent applications tended to improve (i.e., increase) RBC deformability. However, when supra-physiological shear stress was applied, both continuous and intermittent protocols damaged RBC. Changes of RBC mechanical behavior occurred without increases of hemoglobin in the suspending media, thus attesting to the absence of hemolysis., Conclusion: Shear stress has a biphasic effect on the mechanical properties of RBC, with the duration and rate of exposure appearing to have minimal impact on the subhemolytic threshold when compared with the magnitude of applied shear stress.

If a surface is in contact with a solution containing macromolecules or proteins, and the loss of configurational entropy of these molecules at the surface is not balanced by adsorption energy, a polymer-poor layer will develop near the surface. If two such layers overlap, an attractive force develops due to the osmotic pressure difference between these depletion zones and the bulk phase. Recent studies have shown that depletion interaction plays a major role in red blood cell (RBC) aggregation and hence it is a major determinate of blood flow stability; depletion interaction also markedly affects RBC adhesion to vascular endothelial cells. Understanding and quantitating factors that regulate depletion in vivo are thus of importance, yet made difficult since only very small changes of the cell surface (e.g., glycocalyx thickness) such as seen during RBC aging can lead to massive changes of depletion interaction and hence cell-cell adhesion. It is suggested that insight into the in vivo relevance of depletion mechanisms may lead to an improved understanding of how and why blood flow is altered in many diseases, and may also provide new biomarkers (e.g., surface properties) that will aid in the development of novel or improved diagnostic and therapeutic tools.

Cardiovascular diseases (CVD) are the most frequent cause of death throughout the world. The coronary vessel system is a special part of the circulation since there is a continuous change in blood flow, perfusion pressure and shear rate during each cardiac cycle. It is also the place of the narrowest capillaries in the human body, therefore the role of rheological alterations may be of greater importance than in the other parts of the circulatory system. During the past decades, our group has investigated hemorheological parameters (HP) in over 1,000 patients diagnosed with various forms of ischemic heart disease (IHD). In one prospective study, we measured the HP of patients with acute coronary syndrome (ACS). On admission, all examined variables were significantly worse than those of control subjects. During the hospital phase, some of the HP showed further deterioration, and HP remained in the pathologic range during the follow-up period. In another study, we showed that HP are in close correlation with the severity of coronary artery disease. In patients treated with percutaneous coronary intervention, changes in HP were very similar to those observed in subjects with ACS. In a recent study, we analyzed HP in patients undergoing CABG surgery. Our data suggest a hemorheological advantage of off-pump surgery. In another study low Hct/WBV ratio can be regarded as a risk factor of cardiac death in IHD. Our data indicate that rheological parameters are significantly altered in patients with IHD: the extent of the alterations is in excellent correlation with the clinical severity of the disease. Our findings prove that HP play a critical role in the pathogenesis of myocardial ischemia. In recent in vitro and in vivo studies we have investigated the effects of red wine on hemorheological parameters. Our results show that moderate red wine consumption has beneficial effects on hemorheological parameters which may contribute to the French paradox.

Laser-diffraction ektacytometry is a generally accepted technique for measuring RBC deformability induced by fluid shear stress (SS) and yields paired elongation index-SS data at several levels of stress. Unfortunately, comparison of results is hindered by the lack of simple indices that accurately characterize these data. Several mathematical models have been proposed, including those developed for analysis of enzyme kinetics (Lineweaver-Burk, Eadie-Hofstee) and curve fitting (Streekstra-Bronkhorst). All of these analytical approaches provide a value for cell deformation at infinite stress (EImax) and the shear stress required to achieve one-half of this deformation (SS1/2); the use of non-linear regression is essential when calculating these parameters. While the current models provide equivalent results for normal RBC if used with non-linear regression, EImax and SS1/2 are not always concordant for cells with abnormal mechanical behavior. This technical note examines such differences for three conditions: glutaraldehyde treatment, mechanical stress and non-isotonic media. It was found that none of the models yield completely satisfactory values for EImax and SS1/2, especially if there are large changes of EImax. However, the ratio of SS1/2 to EImax (SS1/2/EImax) is much less affected by these problems, has similar power (i.e., standardized difference) as SS1/2 and EImax and is more robust in reflecting alterations of deformability. We thus conclude that the SS1/2/EImax ratio can be used when reporting and comparing various populations of RBC or cells obtained from subjects having different clinical states.

Comparative data on blood composition and blood flow properties indicate different levels of interspecies variation for several parameters. Hematocrit and hemoglobin levels have relatively low variability among mammals, while mean cell volume and red blood cell (RBC) count are more variable. There is also a difference of variability between high and low shear rate blood viscosity in mammals, with low shear rate viscosity having a higher degree of interspecies variation. This observation parallels the diversity of RBC aggregation among mammalian species. Allometric relations for blood rheology parameters indicate highly significant correlations of low shear rate blood viscosity and RBC aggregation with body weight, especially if species belonging to the order Artiodactyla are excluded. These allometric relations can be used to formulate various hypotheses about the evolutionary histories of circulatory functions, as well as hypotheses related to the role of RBC aggregation in the mammalian circulatory system. Such comparative studies and analytical reasoning based on comparative data may contribute to answering the "why" questions, and accordingly may improve our understanding of circulatory functions and hence may have possible clinical importance. During the last several decades, similar approaches to various medical concepts have yielded a new approach to medicine that is now known as Evolutionary Medicine.

Red blood cells (RBC) are exposed to various levels of shear stress (SS) during their flow in the circulatory system, yet no significant damage occurs if their mechanical stability is not impaired. Alternatively, normal RBC may be damaged during flow in non-physiological environments and under extreme SS (e.g., extracorporeal circulation, ventricular assist devices). The shear-induced damage may result in hemolysis or in altered mechanical properties of RBC that, in turn, reduces the ability of RBC to withstand further damage by SS. An ektacytometer employing a Couette shearing system was used to apply SS at a constant level of 100 Pa for 300 seconds as a model of sub-hemolytic mechanical stress. The degree of cellular damage during and after the application was assessed by diffraction pattern analysis. The area of the diffraction pattern was found to correlate with the number of RBC in the sheared suspension. Monitoring the ellipse area during the application of gradually increasing SS provides the concentration of the remaining intact RBC and can therefore be used to estimate the hemolytic threshold as a measure of RBC mechanical stability. The hemolytic threshold determined after the mechanical stress application was found to be ~150 Pa, while it was ~250 Pa in the same samples before the SS application. Additionally, SS-elongation index curves recorded before and after the application of the sub-hemolytic SS significantly differed from each other, indicating the impairment in deformability following the mechanical stress. The Couette type ektacytometer can be used as a tool to assess the sub-hemolytic damage to RBC in testing the biomedical equipment.

Red blood cells (RBC) possess a functional nitric oxide synthase (NOS) enzyme located in the cell membrane and cytoplasm. It has previously been observed that shear stress acting on RBC activates NOS and causes enhanced NO export. The aim of the present study was to investigate the physiological importance (e.g., in local blood flow regulation) of RBC-derived NO stimulated by application of shear stress. Blood samples and arterial vessel segments were obtained from Wistar rats; RBC suspensions were adjusted to a hematocrit of 0.1 l/l using Krebs solution. In order to apply shear stress to the RBC suspensions they were continuously flowed through a small-bore glass tube for 20 minutes at a wall shear stress of 2 Pa. The RBC suspensions were then perfused through endothelium denuded small mesenteric arteries having a diameter of ~300 μm under both high oxygen (PO2 ~130 mmHg) and hypoxic conditions. Perfusion of vessel segments with sheared RBC suspensions caused a significant dilation response under hypoxic conditions but not at high oxygen levels. Incubation of RBC suspensions with the non-specific NOS inhibitor L-NAME (10-3 M) prior to shear stress application abolished this dilation response. Our results indicate that NO released from RBC due to shear stress activation of NOS results in vasodilation of vessel segments under hypoxic conditions, and strongly suggest that NO originating from RBC may have a functional role in local blood flow regulation.

Classically, it is known that red blood cell (RBC) deformability is determined by the geometric and material properties of these cells. Experimental evidence accumulated during the last decade has introduced the concept of active regulation of RBC deformability. This regulation is mainly related to altered associations between membrane skeletal proteins and integral proteins, with the latter serving to anchor the skeleton to the lipid matrix. It has been hypothesized that shear stress induces alterations of RBC deformability: the current study investigated the dynamics of the transient improvement in deformability induced by shear stress at physiologically-relevant levels. RBC were exposed to various levels of shear stress (SS) in a Couette type shearing system that is part of an ektacytometer, thus permitting the changes in RBC deformability during the application of SS to be monitored. Initial studies showed that there is an increase in deformability of the RBC subjected to SS in the range of 5-20 Pa, with kinetics characterized by time constants of a few seconds. Such improvement in deformability, expressed by an elongation index (EI), was faster with higher levels of SS and hence yielded shorter time constants: absolute values of EI increased by 3-8% of the starting level. Upon the removal of the shear stress, this response by RBC was reversible with a slower time course compared to the increase in EI during application of SS. Increased calcium concentration in the RBC suspending medium prevented the improvement of deformability. It is suggested that the improvement of RBC deformability by shear forces may have significant effects on blood flow dynamics, at least in tissues supplied by blood vessels with impaired vasomotor reserve, and may therefore serve as a compensating mechanism for the maintenance of adequate microcirculatory perfusion.

Red blood cell (RBC) aggregate to form two- and three-dimensional structures when suspended in aqueous solutions containing large plasma proteins or polymers; this aggregation is reversible and shear dependent (i.e., dispersed at high shear and reformed at low or stasis). The extent of aggregation is the main determinant of low shear blood viscosity, thus predicting an inverse relationship between aggregation and in vivo blood flow. However, the effects of aggregation on hemodynamic mechanisms (e.g., plasma skimming, Fåhraeus Effect, microvascular hematocrit) may promote rather than impede vascular blood flow. The impact of enhanced RBC aggregation on endothelial function and hemostatic mechanisms adds further complexity, thereby requiring specific attention to the nature, extent and time course of aggregation when considering its overall influence on tissue perfusion. A detailed understanding of aggregation effects is important from a clinical point of view since it may be enhanced during a variety of pathophysiological processes, including infections, circulatory and metabolic disorders, hematological pathologies and several other disease states. Altered RBC aggregation may be an indicator of disease as well as a factor affecting the course of the clinical condition; the prognostic value of RBC aggregation indices has been demonstrated in various diseases. Currently, RBC aggregation is an easily and accurately measurable parameter, and therefore may be expected to have broader clinical usage in the future.

Ektacytometry measures the shape of red blood cells under shear stress by analyzing the diffraction pattern of laser light passing through a thin layer of suspended cells. Here we model the diffraction pattern using a combination of Bessel and anomalous scattering functions, and employ a global pattern-fitting technique over nine different shear stresses to determine the separate mechanical properties of normal and non-deformable cells. This technique is capable of yielding the correct elongation index of the normal cells over a range of shear stresses even when they are mixed with as much as 50% non-deformable cells. Additionally, the relative concentrations of normal and non-deformable cells can be determined.

There is accumulating evidence that exercise may improve disturbed haemorheological parameters that are typically observed in various chronic diseases, thus there is a growing interest in exploring the influence of various exercise models for the improvement of haemorheology. Blood sampling using venipuncture, however, can be limiting during exercise and/or in field settings. The purpose of the present study was to investigate whether venous and capillary blood samples yield comparable red blood cell (RBC) deformability and aggregation indices. Twelve healthy volunteers (6 males and 6 females; age 30 ± 9 yrs; body mass index 24.9 ± 2.8 kg m(-2)) provided blood samples that were collected simultaneously from: i) a prominent forearm vein by venipuncture; ii) the earlobe using a lancet; iii) the middle finger using a lancet. Haematocrit, RBC deformability (Rheoscan-D, Sewon Meditech Inc., Korea) and RBC aggregation (Myrenne GmbH, Roetgen, Germany) were measured for each sample. Haematocrit and RBC deformability were not different between the three sampling sites, and the group averages of RBC aggregation parameters were not different between the three sampling methods. The time course of RBC aggregation was slower when using blood sampled from the earlobe, and there was stronger agreement between RBC aggregation parameters measured using venous and capillary finger samples compared with venous and earlobe. It is suggested that capillary blood sampling from the finger may provide a reliable alternative to venous blood sampling in clinical and field settings.

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.

Nitric oxide (NO) is accepted to be an important factor affecting the degree of vascular tone in various portions of the circulation. Until recently, research in this area has focused on endothelial cells as a NO source, and there is general agreement that: 1) the level of wall shear stress is the primary determinant of endothelial nitric oxide synthase (eNOS) expression; 2) exercise training induces changes of endothelial cell NO synthesizing activity; 3) phosphorylation patterns of eNOS are altered following exercise episodes. However, there is now a growing body of evidence for the existence of similar nitric oxide synthesizing mechanisms in human red blood cells (RBC). Erythrocyte NOS activity has been demonstrated to be induced by applied shear stress and mechanical deformation of RBC, and there are closely linked increases of intracellular nitric oxide levels and of release of NO into the suspending phase. In brief, the RBC is an enzymatic source of NO that is dependent on flow dynamics and from which NO is released in very close proximity to vessel walls. Although reports regarding the influence of exercise on RBC nitric oxide synthesizing mechanisms are not yet concordant, it seems logical to suggest that this source of NO may play a role in the regulation of local blood flow dynamics during exercise.

Prior studies exploring the effects of lanthanides (Ln) on red blood cells (RBC) have primarily focused on ion transport, cell fusion, and membrane protein structure. Our previous report [Biorheology 44 (2007), 361-373] dealt only with lanthanum (La) and cell rigidity; the present study extends these observations to other lanthanides (Nd, Sm, Eu, Dy, Er) and to RBC response to mechanical shear. Deformation-shear stress behavior of normal human RBC was measured at Ln concentrations up to 200 μM. In another series of experiments, RBC were exposed to mechanical stress (190 Pa, 300 s) at 50 μM Ln and deformation-stress data obtained prior to and after this stress. Data were fitted to a Lineweaver-Burke model to obtain the shear stress at one-half maximum deformation (SS1/2). Our results include: (1) lanthanides cause decreased cell deformability with the magnitude of the decrease dependent on concentration and shear stress; (2) this decrease of deformability is affected by Ln ionic radius such that La>Nd>Sm>Eu>Dy>Er and is reversible for cells in Ln-free media; (3) mechanical stress decreases deformability (i.e., increases SS1/2) such that compared to control, La and Sm reduce and Dy and Er enhance the mechanical stress effect; (4) the decrease of deformability consequent to mechanical stress scales inversely with Ln ionic radius. These results indicate a reciprocal relation between cell rigidity and sensitivity to mechanical stress that is mediated by Ln ionic radius. Additional studies are clearly warranted, particularly those that explore membrane-glycocalyx and intracellular mechanisms., (© 2011 – IOS Press and the authors. All rights reserved)

A new method is described in this paper that allows measurement of red blood cell (RBC) aggregation indexes in disposable glass tubes within minutes. Light transmission through the RBC suspension filled into a microhematocrit capillary at stasis is recorded during RBC aggregation; a novel method assures an initial dispersion of aggregates in the capillary. The resulting light transmittance-time data are analyzed to calculate various parameters. Measurement of erythrocyte sedimentation rate (ESR) and RBC aggregation using well established methods and the newly developed capillary tube aggregometer in blood samples with a wide range of RBC aggregation indicated significant correlations between these parameters. Additionally, light transmittance during complete disaggregation allows estimating hematocrit, thereby enabling hematocrit correction of the measured and calculated parameters. The newly developed capillary tube RBC aggregometer is suitable for use as a method to rapidly monitor disease activity and the acute phase response, especially at the point-of-care (e.g., health care facilities, physician's office) and for field studies.

Previous reports have demonstrated that red blood cells (RBC) have an active nitric oxide (NO) synthesizing mechanism which has properties similar to endothelial nitric oxide synthase (eNOS). This red cell NOS activity contributes to the NO export from RBC. The present study explored the influence of shear stress applied to RBC on NO concentrations of cell suspensions. RBC were exposed to shear stress by filtration through 5 microm diameter pores under 10 cm H2O pressure, generating a wall shear stress of approximately 110Pa. NO concentration in the RBC suspensions were measured using electrochemical NO probes before and after filtration through the micropores. NO concentration was found to be significantly increased after a single passage of RBC suspensions through the micropores. The increment in NO concentration depended on the presence of calcium, being 21.8+/-4.4 nM with 1 mM calcium and 13.7+/-2.7 nM without added calcium. Including the calcium chelator EDTA completely abolished this increase. The increment of NO was also affected by the level of oxygenation, being more pronounced under hypoxic conditions. These results confirm that RBC NO generating mechanisms can be stimulated by exposing red cells to shear stress and that calcium plays a role in this stimulation.

The flow properties of blood and its components vary widely throughout the animal kingdom. Even if nucleated avian and reptile red blood cells (RBC) are excluded from the analysis, RBC exhibit different rheological behavior among mammalian species. Both RBC aggregation and cellular deformability have been reported to vary among species, including placental mammals, marsupials, terrestrial and aquatic mammals. Although the relationships between blood flow behavior and species-specific characteristics have not been systematically investigated, studies to date allow recognition of interesting patterns, especially for RBC properties. These properties do not correlate with simple cellular parameters (e.g. mean cell volume), but more detailed analysis of RBC structure may reveal cellular aspects (e.g. surface charge density) that can be related to rheologic behavior. It has been postulated that the athletic capacity of mammalian species may predict the aggregation behavior of their RBC, but this hypothesis has not been supported by data from a wide range of athletic and sedentary species. Aquatic mammals also exhibit a very interesting diversity of hemorheological properties, which again are not yet easily related to specific circulatory adaptations. Data from current comparative studies suggest that a better understanding of the relations between specific hemorheological properties and specific hemodynamic adaptations in a variety of species should contribute to a better understanding of circulatory behavior; future studies are thus clearly indicated.

Sickle cell disease (SCD), a genetically-determined pathology due to an amino acid substitution (i.e., valine for glutamic acid) on the beta-chain of hemoglobin, is characterized by abnormal blood rheology and periods of painful vascular occlusive crises. Sickle cell trait (SCT) is a typically benign variant in which only one beta chain is affected by the mutation. Although both SCD and SCT have been the subject of numerous studies, information related to neurological function and transfusion therapy is still incomplete: an overview of these areas is presented. An initial section provides pertinent background information on the pathology and clinical significance of these diseases. The roles of three factors in the clinical manifestations of the diseases are then discussed: hypoxia, autonomic nervous system regulation and blood rheology. The possibility of a causal relationship between these three factors and sudden death is also examined. It is concluded that further studies in these specific areas are warranted. It is anticipated that the outcome of such research is likely to provide valuable insights into the pathophysiology of SCD and SCT and will lead to improved clinical management and enhanced quality of life.

Blood samples used in hemorheological studies may be stored for a period of time, the effects of storage have yet to be fully explored. This study evaluated the effects of storage temperature (i.e., 4 degrees C or 25 degrees C) and duration on RBC deformability and aggregation for blood from healthy controls and from septic patients. Our results indicate that for normal blood, RBC deformability over 0.3-50 Pa is stable up to six hours regardless of storage temperature; at eight hours there were no significant differences in EI but SS1/2 calculated via a Lineweaver-Burk method indicated impaired deformability. Storage temperature affected the stable period for RBC aggregation: the safe time was shorter at 25 degrees C whereas at 4 degrees C aggregation was stable up to 12 hours. Interestingly, blood samples from septic patients were less affected by storage. Blood can thus be stored at 25 degrees C for up to six hours for deformability studies, but should be limited to four hours for RBC aggregation; storage at 4 degrees C may prolong the storage period up to 12 hours for aggregation but not deformability measurements. Therefore, the time period between sampling and measurement should be as short as possible and reported together with results.

The electrical properties of red blood cell (RBC) suspensions are influenced by flow conditions, and prior studies indicate that electrical properties may reflect the kinetics of RBC aggregation. Changes of conductance and capacitance were monitored and had a time course resembling a "syllectogram" (i.e., temporal change of light reflectance from an RBC suspension after sudden cessation of flow). In the present study, both AC electrical conductance (EC) across and light transmission (LT) through a 1 mm ID glass tube were recorded simultaneously after a sudden stoppage of flow for RBC at various hematocrits in plasma or in isotonic saline (PBS). Preliminary results indicate that EC and LT signals for RBC in plasma have similar time courses, both increasing after an initial decrement of a few seconds duration. Aggregation indexes and aggregation half times calculated using LT and EC showed a similar dependence on hematocrits between 30-50%. Interestingly, RBC in PBS also exhibited a syllectogram time course for conductance, whereas LT continued to decrease after an initial decline reflecting RBC shape recovery. These results suggest that electrical conductance in aggregating and non-aggregating suspensions may be sensitive to phenomena other than RBC aggregation.

It has been previously demonstrated that red blood cells (RBC) possess functional nitric oxide (NO) synthesizing mechanisms. RBC are also equipped with variety of intracellular control mechanisms, and respond to mechanical forces and to various biological stimuli by increased release of ATP. Nitric oxide has also been demonstrated to be released from RBC under certain circumstances, and it has been hypothesized that NO synthase (NOS), which is located in both the RBC membrane and cytoplasm, might be activated by mechanical factors. The present study aimed at investigating NOS activation and NO export induced by mechanical stress applied to RBC in suspension. Heparinized venous blood samples were obtained from healthy, adult volunteers and their hematocrit adjusted to 0.4 l/l. The RBC suspensions were equilibrated at room temperature (22+/-2 degrees C) with either room air or made hypoxic (36 mmHg, approximately 70% saturation) using moisturized 100% nitrogen. The samples were then continuously pumped through a glass tube (diameter = 0.06 cm; length = 33 cm) for 30 min using a dual syringe pump to maintain a wall shear stress of 0.5-2 Pa with NO concentrations in the RBC suspensions measured electrochemically. NO concentration significantly increased under the influence of 2 Pa in hypoxic RBC suspensions: 105.0+/-14.2 nM to 127.1+/-12.0 nM as the peak value at 20 min of perfusion. No increase was observed at lower levels of shear stress. Plasma nitrite/nitrate concentrations were measured in samples obtained at five minute intervals. Application of fluid shear stress to hypoxic RBC suspensions resulted in a significant, time-dependent increase of plasma nitrite/nitrate levels, reaching to 14.7+/-1.5 microM from a control value of 11.2+/-1.3 microM. The presence of the non-specific NOS inhibitor L-NAME (10(-3) M) prevented this increment. Additionally, both eNOS and serine 1177 phosphorylated eNOS immuno-fluorescence staining in RBC cytoplasm were shown to increase in response to applied shear stress. Our results support the hypothesis that RBC NO synthase is activated and that export of NO from RBC is enhanced by mechanical stress.

In December 2008, the International Society for Clinical Hemorheology organized a workshop to evaluate and compare three ektacytometer instruments for measuring deformability of red blood cells (RBC): LORCA (Laser-assisted Optical Rotational Cell Analyzer, RR Mechatronics, Hoorn, The Netherlands), Rheodyn SSD (Myrenne GmbH, Roetgen, Germany) and RheoScan-D (RheoMeditech, Seoul, Korea). Intra-assay reproducibility and biological variation were determined using normal RBC, and cells with reduced deformability (i.e., 0.001-0.02% glutaradehyde (GA), 48 degrees C heat treatment) were employed as either the only RBC present or as a sub-population. Standardized difference values were used as measure of the power to detect differences between normal and treated cells. Salient results include: (1) All instruments had intra-assay variations below 5% for shear stress (SS)>1 Pa but a sharp increase was found for Rheodyn SSD and RheoScan-D at lower SS; (2) Biological variation was similar and markedly increased for SS<3-5 Pa; (3) All instruments detected GA-treated RBC with maximal power at 1-3 Pa, the presence of 10% or 40% GA-modified cells, and the effects of heat treatment. It is concluded that the LORCA, Rheodyn SSD and RheoScan-D all have acceptable precision and power for detecting reduced RBC deformability due to GA treatment or heat treatment, and that the SS range selected for the measurement of deformability is an important determinant of an instrument's power.

Ultrasound characterization of erythrocyte aggregation (EA) is attractive because it is a non-invasive imaging modality that can be applied in vivo and in situ. An experimental validation of the Structure Factor Size Estimator (SFSE), a non-Rayleigh scattering model adapted for dense suspensions, was performed on 4 erythrocyte preparations with different aggregation tendencies. Erythrocyte preparations were circulated in Couette and tube flows while acoustically imaged over a bandwidth of 9-28 MHz. Two acoustically derived parameters, the packing factor (W) and ensemble averaged aggregate size (D), predictably increased with increasing EA, a finding corroborated by bulk viscosity measurements. In tube flow, a "black hole" reflecting the absence of aggregates was observed in the center stream of some parametric images. The SFSE clearly allowed quantifying the EA spatial distribution with larger aggregates closer to the tube walls as the aggregation tendency was increased. In Couette flow, W and D were uniformly distributed across the shear field. Assuming that the viscosity increase at low shear is mainly determined by EA, viscosity maps were computed in tube flow. Interestingly, erythrocyte suspensions with high aggregabilities resulted in homogeneous viscosity distributions, whereas a "normal" aggregability promoted the formation of concentric rings with varying viscosities.

Hematocrit (Hct) is the major determinant of whole blood viscosity and of its oxygen binding capacity: with increasing Hct, viscosity increases exponentially and oxygen capacity increases linearly. Thus, the theoretical oxygen transport potential of blood, as indexed by the ratio of Hct to viscosity (Hct/viscosity), generally yields a curve concave to the Hct axis with a maximum at an "optimal hematocrit" value. This study analyzed relations between Hct, blood viscosity and shear rate for rats and dogs to explore whether different optima exist for Hct or Hct/viscosity. Our results reveal differences depending on both shear rate and species: at equal Hct, rats had higher blood viscosity and thus lower Hct/viscosity levels. Optimum values for Hct/viscosity were markedly different between the two species at shear rates of 90 and 200 s-1. Conversely, Hct/viscosity data at 10 s-1 did not exhibit an optimum but rather a linear decrease of the ratio with increasing hematocrit. Relations between Hct and blood viscosity thus differ among animal species. Inasmuch as animal studies are often utilized as an aid to understanding hemorheological aspects of clinical conditions and/or therapy, evaluating Hct/viscosity ratios may be a useful supplementary tool for research focused on various physiological and patho-physiological processes.

Measurements of red blood cell (RBC) deformability and aggregation can be subject to influence by pre-analytical handling procedures, with the degree of hemoglobin oxygenation having the potential to affect the results. To examine such effects, RBC deformability and aggregation were studied before and after oxygenation or deoxygenation of human blood samples. RBC deformability was assessed using a laser-diffraction ektacytometer having Couette geometry. RBC aggregation was assessed using the same system by monitoring light backscattering after a sudden cessation of high shear; aggregation was also measured by monitoring light transmittance through RBC suspensions. RBC deformability was found to be significantly increased after equilibrating RBC with ambient air (pO2: 142.0+/-3.1 mmHg) compared to the non-oxygenated sample (pO2: 42.4+/-1.8 mmHg). In contrast, equilibration with 100% nitrogen resulted in significant impairment in RBC deformability. RBC aggregation parameters were also affected by oxygenation if measured based on light backscattering, but not if measured using light transmittance. It is thus recommended that blood samples be oxygenated by repeated exposure to ambient air prior to the measurement of hemorheological parameters.

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.

Aerobic performance is dependent on both cardio-respiratory and peripheral factors with hemodynamic parameters playing a major role. However, whether blood rheology might affect aerobic performance through an effect on hemodynamic factors is not known. The aim of the present study was to assess the relationships between hemodynamic, hemorheological and metabolic parameters in response to a sub-maximal cycling exercise protocol consisting of three successive levels of nine min duration (50, 100 and 150 W). Ten young sportsmen participated in the present study. Mean arterial pressure (MAP) was measured manually, with thoracic impedance used to monitor cardiac output (Qc): systemic vascular resistance (SVR) was then calculated. Whole blood viscosity (etab) was measured and used to calculate systemic vascular hindrance. Hematocrit (Hct) was determined by micro-centrifugation and red blood cell (RBC) deformability (EI) was determined by ecktacytometry. A breath-by-breath gas analyzer was used to measure oxygen uptake (VO2); the Fick equation was used to calculate arterio-venous oxygen difference [(a-v)O(2)] from VO(2) and Qc. All measurements were performed at rest, during exercise and during recovery. Compared to baseline, Qc, MAP, Hct, EI, VO(2), and (a-v)O(2) increased during exercise. etab increased above baseline only at 150 W and remained elevated during recovery; the increase in etab during the last level of exercise was associated with a decrease of SVR and systemic vascular hindrance. There was a significant negative correlation between EI and SVR (r=-0.35, p<0.01) and a significant positive relationship between EI and (a-v)O(2) (r=0.35, p<0.01) and between EI and VO(2) (r=0.37, p<0.01) across all exercise workloads, thus suggesting a potential role for RBC deformability as a factor affecting aerobic performance via oxygen delivery to tissues. These data lend support to the concept that hemorheological parameters may contribute to hemodynamic and cardio-respiratory adaptations in response to exercise in moderately trained sportsmen.

We have recently demonstrated that the use of a tourniquet to locate the vein prior to blood sampling may affect red blood cell (RBC) deformability and aggregation, even if the tourniquet is quickly removed after insertion of the needle into the vein. However, in clinical practice, the tourniquet is usually kept in place until blood sampling is completed. The present study tested the effects of tourniquet application during the blood sampling period on RBC deformability and aggregation. We compared blood samples obtained at 5, 30, 60 and 90 seconds following the tourniquet application; a control blood sample was obtained without applying a tourniquet. RBC deformability was slightly increased at 5 seconds (7.6% mean, 1-5 Pa) and at 60 seconds (5.9% mean, 1-30 Pa) but not at 30 or 90 seconds, with the increase inversely related to shear stress. The RBC aggregation index was slightly but significantly decreased at 5, 30 and 60 seconds, with no changes observed for either amplitude or half-time of aggregation. These very modest yet significant effects of tourniquet application on hemorheological parameters may not be meaningful in terms of pathophysiology, but may become important in data interpretation or for specific subjects.

The pathophysiological abnormalities of stable angina (SA) and acute coronary syndromes (ACS) may, in part, be promoted by fluid forces associated with local blood flow and hence by the rheological properties of blood. This study evaluated several hemorheological parameters in 16 healthy controls and in 16 SA, 18 unstable angina (UA) and 19 acute myocardial infarct (AMI) patients; all patients underwent diagnostic angiography following blood sampling. Rheological measurements included whole blood viscosity, plasma viscosity and RBC aggregation via erythrocyte sedimentation rate (ESR) and Myrenne aggregometer indices. Compared to controls, RBC aggregation was significantly elevated in all patient groups (p<0.001), with the rank being AMI>UA>SA. RBC aggregability as tested in 70 kDa dextran exceeded control in all patients. Blood viscosity values calculated at 40% Hct, plasma viscosity and yield shear stress values followed the same pattern (AMI>UA>SA>control); increases of inflammatory markers (i.e., WBC count, hs-CRP) were elevated in all patient groups in the order AMI>UA>SA. Our study thus indicates an association between hemorheological abnormalities and the severity of coronary artery disease, and suggests the merit of evaluating whether therapeutic interventions that normalize blood rheology may reduce the incidence and/or progression of coronary artery disease.

Prior reports describing the effects of lanthanum (La(3+)) on red blood cells (RBC) have focused on the effects of this lanthanide on cell fusion or on membrane characteristics (e.g., ion movement across membrane, membrane protein aggregation); the present study explores its rheological and biophysical effects. Normal human RBC were exposed to La(3+) levels up to 200 microM then tested for: (1) cellular deformability using a laser-based ektacytometer and an optical-based rheoscope; (2) membrane viscoelastic behavior via micropipettes; (3) surface charge via micro electrophoresis. La(3+) concentrations of 12.5 to 200 microM caused dose-dependent decreases of deformability that were greatest at low stresses: these rheological changes were completely reversible upon removing La(3+) from the media either by washing with La(3+)-free buffer or by suspending La(3+)-exposed cells in La(3+)-free media (i.e., viscous dextran solution). Both membrane shear elastic modulus and membrane surface viscosity were increased by 25-30% at 100 or 200 microM. As expected, La(3+) decreased RBC electrophoretic mobility (EPM), with EPM inversely but not linearly associated with deformability; changes of EPM were also completely reversible. These results thus indicate novel aspects of RBC cellular and membrane rheological behavior yet raise questions regarding specific mechanisms responsible for La(3+)-induced alterations.

Although the study of red blood cell (RBC) aggregation continues to be of basic science and clinical interest, aggregation standards for calibration do not exist, and most aggregation studies report data in terms of arbitrary units: quantitative comparisons between studies are thus essentially precluded. However, use of low shear viscometry plus the Casson equation provides a yield shear stress that has defined units and is known to reflect RBC aggregation. Employing human RBC-plasma suspensions exhibiting a wide range of aggregation, the present study examined relations between yield shear stress values and aggregation indices obtained using the Myrenne aggregometer: the latter approach uses a light-transmission technique and provides an "M" index at stasis and an "M1" at very low shear. Our results for normal controls and for angina patients without coronary artery disease indicate highly significant correlations (p<0.001) between the yield stress and both M and M1. Thus, within the range of aggregation studied, these findings lend support to the rheological validity of the Myrenne approach; extension of our findings to intensely aggregating RBC suspensions may require additional validation studies.

Changes in hemorheological parameters were studied in dogs following unilateral renal artery clamping (45-minute ischemia then reperfusion), with and without preoperative administration of allopurinol. Sham-operated animals were also evaluated. Blood samples were collected preoperatively, at beginning and at 30, 60 and 120 minutes of reperfusion, then on the 1st, 3rd, 5th and 7th days. Filtration properties of erythrocytes (relative cell transit time, RCTT), whole blood and plasma viscosity (WBV, PV), fibrinogen level and hematology parameter were determined. RCTT significantly increased for both ischemic groups at 30 minutes of reperfusion, and remained elevated on the 1st and 2nd postoperative days; these changes were abolished by allopurinol pretreatment. WBV and hematocrit increased on the 1st day, and PV and fibrinogen level showed elevation on 1st-5th postoperative days. We thus conclude that decreases of RBC deformability (i.e., higher RCTT) were characteristic and specific on early postoperative days after renal ischemia-reperfusion and that these alterations were prevented by pre-ischemia administration of allopurinol.

3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are the most commonly used cholesterol-lowering drugs, with recent clinical trends usually aimed at achieving the lowest possible plasma cholesterol levels. Although the effects of increased plasma cholesterol have been previously reported, it is not obvious how very low plasma cholesterol levels would affect membrane composition and the deformability of red blood cells (RBC). The present study investigated the effects of hypocholesterolemia achieved by atorvastatin therapy on RBC membrane and mechanical properties in guinea pigs fed a normal diet. Two groups of animals were used (atorvastatin-treated, n=12; control n=12), and atorvastatin given orally in isotonic phosphate-buffered saline (PBS) at a dose of 20 mg/kg/day for a 21-day period. Our results indicate that the atorvastatin-treated group had significantly lower plasma total cholesterol (17.42+/-1.70 mg/dl), low-density lipoprotein cholesterol (5.25+/-2.22 mg/dl) and triglycerides (42.60+/-3.78 mg/dl) than the control group (34.08+/-1.72, 21.17+/-1.41 and 60.64+/-2.43 mg/dl, respectively). In addition, membrane cholesterol content was lower (p<0.0001) and phospholipid content higher (p<0.0001) in the atorvastatin-treated group, thus decreasing the cholesterol to phospholipid ratio; a significant enhancement in sodium-potassium-ATPase activity also occurred. However, in spite the marked changes of plasma and RBC membrane composition, there was no change of RBC deformability. Note that although our results indicate no adverse rheological alterations, extension of our findings to humans requires caution.

Cyclic oligomers of glucose, termed cyclodextrins (CDs), can contain 6 (alpha-CD), 7 (beta-CD) or 8 (gamma-CD) glucose units and are able to remove cholesterol from platelet membranes and decrease platelet aggregation. The present study was designed to examine the effects of these CDs on RBC aggregation and blood viscosity. Blood from normal adult volunteers was incubated at 37 degrees C with 3.0 x 10(-4) to 1.5 mM levels of the CDs, then processed to obtain platelet-rich plasma, platelet poor plasma and 40% hematocrit blood; measurements included collagen-induced platelet aggregation, RBC aggregation (Myrenne Aggregometer) and blood viscosity at 1-1000 sec(-1)(Rheolog). Our results indicate the expected dose-dependent inhibition of platelet aggregation by beta-CD, with no significant effects of alpha-CD or gamma-CD. RBC aggregation studies showed no effect of alpha-CD but highly significant (p<0.01) decreases by both beta-CD and gamma-CD; at the concentrations studied (1.5 x 10(-3) to 1.5 mM), beta-CD had somewhat greater effects. Blood viscosity was not affected by alpha-CD, but was significantly decreased in a dose-dependent manner by beta-CD and, at the highest concentration (1.5 mM), by gamma-CD. Interestingly, the effects of beta-CD and gamma-CD were independent of shear, with these effects not explained by the usual mechanisms. These results suggest the potential hemorheological value of CDs, yet also indicate the need for additional studies.

Knowledge of the effects of red blood cell aggregation on blood flow in small vessels is crucial to a better understanding of resistance changes in the venous microcirculation. Recent studies on rat spinotrapezius muscle indicate that enhanced red blood cell aggregation, induced by dextran 500, significantly affects velocity profiles at pseudoshear rates (the ratio of mean velocity to diameter) less than 40 s(-1). Since the use of a power-law model to describe these profiles does not provide a consistent rheological description, we have evaluated using the Casson model that has been widely used to characterize in vitro blood rheology. In the present study, we report experimental values of rat blood viscosity in the presence of dextran 500 and combine these in vitro measurements with previously obtained in vivo venular velocity profiles to determine whether the Casson model can provide a valid description of in vivo velocity profiles. Our analysis shows that the two-phase Casson model with a peripheral plasma layer is in quantitative agreement with experimentally obtained velocity profiles obtained in venules of rat spinotrapezius muscle under low flow rate. These results have implications for pathological low-flow conditions, such as hemorrhage and sepsis, and they quantitatively describe blunted velocity profiles and elevated flow resistance in postcapillary venules.

The rheologic characteristics of blood and its formed elements continue to be of basic science and clinical interest, with numerous publications dealing with topics such as blood and plasma viscosity, RBC aggregation and cell deformability. Alterations of blood's rheologic behavior in pathologic states have been extensively studied, with the findings usually indicating changes assumed to be detrimental to tissue perfusion (e.g., increased RBC aggregation). However, the current literature contains relatively few studies dealing with two important areas: (1) relations between altered rheologic behavior and in vivo hemodynamics; (2) the effects of therapy in those clinical states associated with altered rheologic behavior. This paper presents brief and selective comments on the assumed importance of two rheologic "abnormalities" (i.e., increased plasma viscosity, increased RBC aggregation), and on whether clinical therapy in diabetes, hypertension and sepsis leads to normalization of the "abnormalities". Finally, a few gratuitous comments are presented regarding possible future directions in the field of hemorheology and hemodynamics.

It is well known that the hematocrit in microvessels with diameters smaller than 1000 microm is lower than either venous or arterial hematocrit, thereby resulting in significantly lower mean hematocrit values for vessels perfusing a given tissue (i.e., lower tissue hematocrit). The mechanisms that underlie this reduction of microvascular hematocrit include axial migration, plasma skimming and the Fahraeus Effect. It has been previously demonstrated in rats that a linear hematocrit gradient normally exists through the thickness of the left ventricular myocardium, and that this gradient is sensitive to alterations of the rheological properties of the circulating blood. The gradient is abolished if the RBC in the perfusate are rigid; fibrinogen infusions, and thus increases of both plasma viscosity and RBC aggregation, also affect this gradient. In a new series of studies, it has been observed that enhanced RBC aggregation affects the myocardial hematocrit gradient regardless of alterations of plasma viscosity. Although the exact mechanisms responsible for the myocardial hematocrit gradient, as well as its physiological significance, are not yet clearly known, it is possible to speculate that alterations in local hematocrit could adversely affect myocardial perfusion and function.

Red blood cell (RBC) aggregation and blood viscosity are important determinants of in vivo blood flow dynamics and, in marine mammals, these parameters may impact diving physiology by altering blood oxygen delivery during the diving response. Weddell seals are superb divers and exhibit age-related patterns in blood oxygen chemistry and diving ability. By contrast, bowhead whales are not long duration divers, and little is known of their blood properties relative to diving. The present study was designed to compare rheological characteristics of blood from Weddell seal pups, Weddell seal adults, and from adult bowhead whales: blood viscosity and RBC aggregation in plasma and in polymer solutions (i.e., RBC "aggregability") were measured. Salient findings included: (1) significant 4- to 8-fold greater aggregation in blood from adult seals compared with pups and human subjects; (2) 2-to 8-fold greater aggregation in bowhead whale blood compared with human blood; (3) compared to human red cells, enhanced RBC aggregability of RBC from adult seals and whales as determined by their greater aggregation in polymer solutions; (4) increasing RBC aggregation and aggregability of seal pup blood over a seven day period following birth; (5) significantly greater blood viscosity for adult seals compared with pups at both native and standardized hematocrits. These results indicate that, for both species, hemorheological parameters differ markedly from those of humans, and suggest progressive changes with seal age; the physiological implications of these differences have yet to be fully defined.