Viscoelastic behavior of erythrocyte membrane.

A nonlinear viscoelastic relation is developed to describe the viscoelastic properties of erythrocyte membrane. This constitutive equation is used in the analysis of the time-dependent aspiration of an erythrocyte membrane into a micropipette. Equations governing this motion are reduced to a nonlinear integral equation of the Volterra type. A numerical procedure based on a finite difference scheme is used to solve the integral equation and to match the experimental data. The data, aspiration length vs. time, is used to determine the relaxation function at each time step. The inverse problem of obtaining the time dependence of the aspiration length from a given relaxation function is also solved. Analytical results obtained are applied to the experimental data of Chien et al. 1978. Biophys. J. 24:463-487. A relaxation function similar to that of a four-parameter solid with a shear-thinning viscous term is proposed.     

Mechanical stresses in erythrocyte membranes (theoretical models)

The type of mechanical stresses that arise in erythrocyte membranes on exposure to catecholamines and steroid hormones is considered. Tensors of mechanical stresses and displacements were obtained for a membrane interacting with hormones. A possible mechanism of membrane rupture under mechanical stresses is discussed. Catecholamines and androgens increase the microviscosity of membranes, and alternating kink and stretching sites occur in the lipid membrane bilayer to produce a checker-wise pattern. The membrane becomes thinner in a stretching site (smectic A → smectic C transition). When tensile stresses increase further and exceed a certain critical value the membrane may rupture. It is possible that a gel phase L_β? → liquid crystalline phase L_α transition takes place in the stretching site of the lipid bilayer prior to disruption. The density of the lipid bilayer decreases in the process, pores form, and then cracks occur.     

Mechanical behavior of fibroblasts included in collagen lattices

Striae distensae are characterized by linear, smooth bands of atrophic-appearing skin. Excessive steroid activity, genetic and mechanical factors and inherited defects of connective tissues are the most frequent causes of this disease. Fibroblasts derived from women presenting striae distensae lesions were included into collagen gels to study their mechanical behavior: capacity to contract free-floating lattices and to produce isometric force in tense lattices. To measure the retracted lattice diameter, the culture dishes were placed on a transparent metric scale. An isometric force system was used to study quantitatively the forces developed during lattice contraction. alpha 2 beta 1 integrins expression (transmembrane receptors) was evaluated by flux cytometry. Striae distensae fibroblasts contract collagen gels slower than normal human fibroblasts but the final contraction is similar. They produce a greater isometric force which is associated with enhanced alpha 2 beta 1 integrins expression. By their mechanical properties, striae distensae fibroblasts appear as a different population from normal fibroblasts.     

Study of erythrocyte sedimentation behavior by piezoelectric crystal impedance sensor

Based on the impedance characteristic of erythrocytes at high frequency, the response of piezoelectric crystal impedance (PCI) sensor in the erythrocyte suspension was derived and verified experimentally. A method of using PCI sensor to investigate erythrocyte aggregation-sedimentation phenomenon was proposed. From the frequency response of the PCI sensor, the erythrocyte aggregation time and sedimentation rate could be obtained during erythrocyte aggregation and sedimentation. With the present method, the effects of the erythrocyte deformability, the osmotic pressure and the coexisting macromolecules on the erythrocyte sedimentation rate were studied. The results show that the PCI sensor possesses some advantages, such as good sensitivity, simplicity of use and no thermal effect for the impedance study of erythrocyte aggregation and sedimentation.     

The behavior of the human erythrocyte as an imperfect osmometer: a hypothesis

The human erythrocyte does not behave as a perfect osmometer that is its volume does not change as predicted with the change of the tonicity of the medium, as if there was a fraction of the cell water not participating in the osmotic exchange. A mechanism of control of the erythrocyte shape has been previously proposed in which Band 3 (AE1), the protein anion exchanger of Cl(-) and HCO(3)(-), plays a central role. Specifically, decrease and increase of the ratio of its outward-facing conformation and inward-facing conformation (Band 3(o)/Band 3(i)) contract and relax the membrane skeleton, thus favoring echinocytosis and stomatocytosis, respectively. The equilibrium Band 3(o)/Band 3(i) ratio is determined by the Donnan equilibrium ratio of anions and protons, increasing with it (r=Cl(i)(-)/Cl(o)(-)=HCO 3(i)(-)/HCO 3(o)(-)=H(o)(+)/H(i)(+)). The Donnan ratio is influenced by the erythrocyte transport and metabolic activities. The volume change of the human erythrocyte alters the skeleton conformation as it is accompanied by a change of the membrane curvature. Thus, the mechanism could be a hypothesis for explaining the behavior of the human erythrocyte as an imperfect osmometer since the Donnan ratio controls the Band 3(o)/Band 3(i) ratio which controls the volume by a control of the degree of contraction or relaxation of the skeleton. Predictions made by the hypothesis on the Ponder's coefficient R' values in the presence of sucrose or Band 3 substrates slowly transported as well as on the participation of Band 3 in the osmotic hemolysis appear to be corroborated by previous observations. If the hypothesis was valid, it would follow that there is a pressure gradient across the erythrocyte membrane. The equilibrium volume is antagonistically determined by the Donnan ratio per se and Band 3. Band 3, rather than the ratio of surface-to-volume, primarily controls the osmotic hemolysis.     

Mechanical implications of chimpanzee positional behavior.

Mechanical hypotheses concerning the function of chimpanzee anatomical specializations are examined in light of recent positional behavior data. Arm-hanging was the only common chimpanzee positional behavior that required full abduction of the humerus, and vertical climbing was the only distinctive chimpanzee positional behavior that required forceful retraction of the humerus and flexion of the elbow. Some elements of the chimpanzee anatomy, including an abductible humerus, a broad thorax, a cone-shaped torso, and a long, narrow scapula, are hypothesized to be a coadapted functional complex that reduces muscle action and structural fatigue during arm-hanging. Large muscles that retract the humerus (latissimus dorsi and probably sternocostal pectoralis major and posterior deltoid) and flex the elbow (biceps brachii, probably brachialis and brachioradialis) are argued to be adaptations to vertical climbing alone. A large ulnar excursion of the manus and long, curved metacarpals and phalanges are interpreted as adaptations to gripping vertical weight-bearing structures during vertical climbing and arm-hanging. A short torso, an iliac origin of the latissimus dorsi, and large muscles for arm-raising (caudal serratus, teres minor, cranial trapezius, and probably anterior deltoid and clavicular pectoralis major) are interpreted as adaptations to both climbing and unimanual suspension.     

Nitric oxide in the lymphatic microvessel regulation

Topical application of sodium nitroprusside on rat mesentery has a marked influence on lymph microvessels function. The drug causes a dilation of majority of lymphangions and decrease of the pacemaker activity of the vessel wall structures and valves. These changes do not lead to lymph stasis, and lymph flow velocity does not reduce. The non-selective inhibitor of NO synthase (N-nitro-L-arginine) intensifies vasomotions of lymph microvessels, modulates phasic contractile activity and increases lymph flow velocity. There is a time dependent dynamic of changes in action of N-nitro-L-arginine. During inhibition of endogenous NO synthesis the responses of lymph microvessels on sodium nitroprusside application are modified: the constriction of majority lymphangions and activation of valve work are observed.     

Solubility behavior of enzymes after addition of polyethylene glycol to erythrocyte hemolysates

The addition of polyethylene glycol to a hemolysate of rat erythrocytes reduces the solubility of phosphofructokinase and glucose-6-phosphate and 6-phosphogluconate dehydrogenases in an exponential manner with respect to polymer concentration. Analyses of the solubility curves (log solubility versus polymer concentration) obtained at different pH values suggest that the solubility can be related to both the aggregation state and the intrinsic solubility of the proteins promoted by solution conditions. These findings suggest the possibility of using polyethylene glycol in a rational way for the fractional precipitation of a mixture.     

Dynamic and electrokinetic behavior of erythrocyte membrane in diabetes mellitus and diabetic cardiovascular disease

The dynamic and electrokinetic properties of erythrocyte membrane are explored as significant indices involved in the association of diabetes and diabetic cardiovascular disease. Lipid peroxidation studies reveal malondialdehyde concentration to reach a maximum in diabetic cardiovascular patients. Lower fluidity of erythrocyte membrane implies declined ability of erythrocyte to deform in pathogenic state, which is supported by decreased osmotic resistance. Membrane protein profile modification detected by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) indicates a significant reduction in the quantity of ankyrin protein band 2.1 in diabetic subjects. In addition the reduction in an immunoreactive band against polyclonal anti-ankyrin antibody during Western blot analysis confirms the modification of ankyrin protein in diseased erythrocyte (reported for the first time). The electrokinetic behavior of erythrocyte membrane is monitored by laser Doppler velocimetry mode of the Nano-ZS. Changes in zeta potential values of the red blood cell membrane are consistent with decreased membrane fluidity in diseased erythrocytes (reported for the first time). Membrane potential values of control, diabetic and diabetic cardiovascular erythrocytes are -37.24+/-1.5 mV, -28.44+/-1.34 mV, and -22.21+/-1.21 mV respectively indicating a gradual lowering of zeta potential when erythrocyte membrane undergoes progressive changes - from simple agglomeration to fluid gel formation - and finally to a rigid gel.     

Different behavior of ghost-linked acidic and neutral sialidases during human erythrocyte ageing

Acidic and neutral sialidases (pH optimum 4.7 and 7.2, respectively) were assayed on human circulating erythrocytes during ageing. The assays were performed on intact erythrocytes and resealed erythrocyte ghost membranes. From young to senescent erythrocytes the acidic sialidase featured a 2.7-fold and 2.5-fold decrease in specific activity when measured on intact cells or resealed ghost membranes, whereas the neutral sialidase a 5-fold and 7-fold increase, respectively.The Ca²⁺-loading procedure was employed to mimic the vesiculation process occurring during erythrocyte ageing. Under these conditions the released vesicles displayed an elevated content of acidic sialidase, almost completely linked through a glycan phosphoinositide (GPI) anchor but no neutral sialidase activity, that was completely retained by remnant erythrocytes together with almost all the starting content of sialoglycoconjugates. The loss with vesiculation of acidic sialidase with a concomitant relative increase of neutral sialidase was more marked in young than senescent erythrocytes.The data presented suggest that during ageing erythrocytes loose acidic sialidase, and get enriched in the neutral enzyme, the vesiculation process, possibly involving GPI-anchors-rich membrane microdomains, being likely responsible for these changes. The enhanced neutral sialidase activity might account for the sialic acid loss occurring during erythrocyte ageing.     

Mechanical action of the blood onto atheromatous plaques: influence of the stenosis shape and morphology

The vulnerability of atheromatous plaques in the carotid artery may be related to several factors, the most important being the degree of severity of the endoluminal stenosis and the thickness of the fibrous cap. It has recently been shown that the plaque length can also affect the mechanical response significantly. However, in their study on the effect of the plaque length, the authors did not consider the variations of the plaque morphology and the shape irregularities that may exist independently of the plaque length. These aspects are developed in this paper. The mechanical interactions between the blood flow and an atheromatous plaque are studied through a numerical model considering fluid–structure interaction. The simulation is achieved using the arbitrary Lagrangian–Eulerian scheme in the COMSOL TM commercial finite element package. The stenosis severity and the plaque length are, respectively, set to 45% and 15 mm. Different shapes of the stenosis are modelled, considering irregularities made of several bumps over the plaque. The resulting flow patterns, wall shear stresses, plaque deformations and stresses in the fibrous cap reveal that the effects of the blood flow are amplified if the slope upstream stenosis is steep or if the plaque morphology is irregular with bumps. More specifically, the maximum stress in the fibrous cap is 50% larger for a steep slope than for a gentle slope. These results offer new perspectives for considering the shape of plaques in the evaluation of the vulnerability.     

Mechanical behavior of coronary stents investigated through the finite element method

Intravascular stents are small tube-like structures expanded into stenotic arteries to restore blood flow perfusion to the downstream tissues. The stent is mounted on a balloon catheter and delivered to the site of blockage. When the balloon is inflated, the stent expands and is pressed against the inner wall of the coronary artery. After the balloon is deflated and removed, the stent remains in place, keeping the artery open. Hence, the stent expansion defines the effectiveness of the surgical procedure: it depends on the stent geometry, it includes large displacements and deformations and material non-linearity.In this paper, the finite element method is applied (i) to understand the effects of different geometrical parameters (thickness, metal-to-artery surface ratio, longitudinal and radial cut lengths) of a typical diamond-shaped coronary stent on the device mechanical performance, (ii) to compare the response of different actual stent models when loaded by internal pressure and (iii) to collect suggestions for optimizing the device shape and performance.The stent expansion and partial recoil under balloon inflation and deflation were simulated. Results showed the influence of the geometry on the stent behavior: a stent with a low metal-to-artery surface ratio has a higher radial and longitudinal recoil, but a lower dogboning. The thickness influences the stent performance in terms of foreshortening, longitudinal recoil and dogboning.In conclusion, a finite element analysis similar to the one herewith proposed could help in designing new stents or analyzing actual stents to ensure ideal expansion and structural integrity, substituting in vitro experiments often difficult and unpractical.     

Mechanical behavior of the fingertip in the range of frequencies and displacements relevant to touch

It was previously suggested that the mechanical properties of the fingertip could be characterized by elasticity from dc to about 100Hz and by viscosity above this frequency. Using a specifically designed high mobility probe, we accurately measured the impedance of the fingertips of seven participants under a variety of conditions relevant to purposeful touch. Direct measurements vindicated previous indirect observations. We also characterized the dependency of the fingertip impedance upon normal load, orientation, and time.     

Use of fluorescence to determine the effects of cholesterol on lipid behavior in sphingomyelin liposomes and erythrocyte membranes

The purpose of this study was to generate the equivalent of a cholesterol/temperature phase map for a biological membrane using fluorescence spectroscopy. The pseudo-phase map was created using human erythrocytes treated with various concentrations of methyl-beta-cyclodextrin to remove defined amounts of cholesterol and a trio of fluorescent probes that assess different membrane properties (laurdan, diphenylhexatriene, and merocyanine 540). Parallel experiments with two-photon microscopy suggested that changes in cellular cholesterol content affected the entire membrane rather than being localized to specific macroscopic domains. The various regions of the composite erythrocyte pseudo-phase map were interpreted using analogous data acquired from multilamellar vesicles that served as simplified models of cholesterol-dependent phases. The vesicles consisted of various concentrations of cholesterol (0 to 50 mol%) with either palmitoyl sphingomyelin, 1:1 dipalmitoylphosphatidylcholine and dioleoylphosphatidylcholine, or phospholipid mixtures intended to simulate either the inner or outer leaflet of erythrocyte membranes. Four distinguishable regions were observed in sphingomyelin phase maps corresponding to the traditional solid-ordered and liquid-disordered phases and two types of liquid-ordered behavior. Physical properties were less diverse in the mixed phospholipid vesicles, as expected, based on previous studies. Erythrocytes displayed five regions of different combinations of membrane properties along the phase map. Some of the observations identified similarities between the cells and liquid-ordered behavior observed in the various types of liposomes as well as some interesting differences.     

Thermotropic behavior of intact human erythrocyte membranes detected by measuring their surface conductivity

A relative surface conductivity Hz/Hm of intact human erythrocytes was investigated by scanning the temperature interval 5-55 degrees C. The form of thus obtained thermogram implies a relation between Hz/Hm and thermotropic behavior of the erythrocyte membrane in vivo. Conditions for the best manifestation of this relation were studied.