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Biomedical subjects

G B Nash

Publications and source records attributed to G B Nash.

At least 91 records · Page 5Linked to original sources
PubMed1991

Red cell mechanics: what changes are needed to adversely affect in vivo circulation.

The ability of red cells to deform is essential to allow their circulation. However the degree of rheological abnormality which can be tolerated before flow is impaired is not so clear. Red cell rheology has been characterised in a number of physiological, pathological and genetic conditions, and some inferences can be drawn. In vivo aging causes a small loss of cell deformability attributable to increased membrane and internal viscosity; volume and surface area are also lost. These changes cannot be sufficient to cause cellular removal, since the cells sampled had continued to circulate. In sickle cell disease, the oxygenated blood contains dense cells that are more severely abnormal than dense, aged cells from normal individuals. Melanesian ovalocytes have comparable rigidity to dense SS cells, but this condition has no marked circulatory pathology. Thus circulatory problems in SS disease probably stem from deoxygenation-induced sickling which causes extreme loss of deformability, rather than from the abnormal cells in oxygenated blood. In falciparum malaria, immature parasites cause appreciable loss of deformability but continue to circulate. Maturation of the parasites causes much greater rheological changes, including attachment to vascular endothelium, and the cells cease to circulate. In summary, quite marked changes in cell mechanics can occur without loss of ability to circulate. It thus seems that slight rheological alterations reported in some clinical studies are unlikely to cause appreciable flow disruption.

Biomechanical Phenomena↗
PubMed1991

Effect of dehydration on the viscoelastic behavior of red cells.

Dehydration of red cells alters their rheological behavior and may contribute to the pathology of disorders such as sickle cell disease. We have measured the viscoelastic properties of individual human HbAA red cells after graded dehydration induced by incubation with valinomycin at different external K levels. With dehydration, the cells underwent progressive reduction in their rate of extensional shape recovery (i.e., after elongation by micropipette manipulation). Their rigidity remained unaffected until the mean cell hemoglobin concentration (MCHC) rose above 50 g/dl, but then increased about 100% as judged from the response of membrane tongues drawn into micropipettes. There was also a marked reduction in the shape recovery rate at this level of dehydration, and the cells no longer behaved elastically but rather showed unrecoverable residual deformation. Additionally, the cytoplasm took on solid-like properties. Changes in cell rigidity and shape recovery rate have been previously demonstrated for dense sickle cells; our results indicate that normal red cells can be induced to behave similarly, but that a greater degree of dehydration is required.

Adult↗
PubMed1991

Platelet-induced granulocyte aggregation in vitro.

Recent studies have shown that the response of granulocytes to stimulation is increased by the presence of platelets. Using an assay based on counting aggregates with a Coulter Counter, we found that heparinized or citrated platelet-rich plasma directly caused aggregation of isolated granulocytes. Platelets themselves were frequently present within the granulocyte aggregates. The degree of aggregation depended on the platelet:granulocyte ratio, and increased over a 2--5 min period of mixing, with a slower increase thereafter. Aggregation was calcium-dependent, increased by stimulation of the platelets with ADP or collagen but not serotonin, and only slightly reduced by a thrombin inhibitor (hirudin, at concentrations up to 0.9 U/ml). A similar adhesive interaction in vivo might be relevant to thrombotic and ischaemic pathology.

Blood Platelets↗
PubMed1991

Therapeutic aspects of white blood cell rheology in severe ischaemia of the leg.

In order to investigate the involvement of white blood cells (WBC) in ischaemia, we have used micropore filtration techniques to measure the flow properties of cells from patients with severe leg ischaemia, before and after therapy (amputation or pharmacological). Compared to age-matched controls, WBC from patients had impaired ability to flow through 8 microns and 5 microns pore filters. This applied to fractionated granulocytes and mononuclear cells, as well as to unfractionated mixed WBC. WBC from blood drawn from the ischaemic leg had worse filterability than those from arm blood of the same patients. Judging from WBC morphology, the percentage of active WBC was higher in samples drawn from ischaemic patients compared to controls. After amputation of the ischaemic leg, significant improvement was observed, so that the WBC were no longer significantly different from controls. The flow abnormalities may reflect activation of WBC by factors released in the ischaemic tissue. A cycle of WBC trapping, activation, tissue damage and further activation and trapping could contribute to the worsening of tissue ischaemia. Pharmacological intervention in this degenerative cycle might be possible, and our preliminary data show that Trental infusion (600 mg over 6 hours) improves the filterability of granulocytes from severely ischaemic patients.

Arterial Occlusive Diseases↗
PubMed1991

Increased white cell trapping in the dependent legs of patients with chronic venous insufficiency.

Interest in white cell (WBC) rheology has largely been directed towards hypotensive conditions. However, as capillary flow is dependent on the relative arteriovenous pressure gradient, we investigated the dynamics of WBC flow in patients with venous hypertension. Three groups of matched subjects were studied: normal controls, patients with primary varicose veins, and patients with deep venous insufficiency confirmed by foot volumetry and Doppler studies. A venous cannula inserted at the level of the medial malleolus allowed blood samples to be taken at time 0 (supine) and then at 15, 30, 45, and 60 minutes after sitting up. Further samples were taken after subjects resumed a supine position. After 60 minutes sitting the haematocrit of blood leaving the foot increased by 9.6% in controls and 25.6% in patients with venous insufficiency. The difference in the behaviour of WBC was more marked. Controls showed a 5.0% decrease in the relative number of WBC after 60 minutes sitting compared to 28% decrease in patients with venous insufficiency. On resuming a supine position there was a significant increase in the number of WBC leaving the foot and this "wash-out" appeared to be delayed in the two patient groups. Platelets showed a corresponding fall when the foot was dependent but there appeared to be no washout after elevation.

Chronic Disease↗
PubMed1990

Effect of magnesium ions on red cell membrane properties.

Spectrin forms aggregates in solution when incubated at relatively high concentrations (several millimolar) of divalent cations. According to the evidence of electron microscopy, aggregates of globular appearance and rather uniform size are cooperatively formed from spectrin dimers, no intermediate structures being seen. Inter-dimer chemical cross-linking of spectrin in intact red cell membranes is enhanced if magnesium ions at a concentration of 0.5 mM or more are present. On the other hand, the elimination of magnesium from the interior of intact cells causes no significant change in shear elastic modulus, measured by micropipette assays, nor is there any dependence of membrane filtration rate on intracellular free magnesium concentration in the range 0-1 mM. Magnesium-depleted cells are, however, converted into echinocytes within a short period, in which control cells, exposed to ionophore and external magnesium ions, remain completely discoid. Magnesium-depleted cells also undergo structural changes on heating below the temperature at which vesiculation sets in. These reveal themselves by the transformation of the cells to a unique characteristic shape, by grossly reduced filtrability, and by extensive agglutination of the cells when treated with a bifunctional reagent. Magnesium ions thus regulate the stability, but not to any measurable extent the gross elasticity, of the red cell membrane.

Cross-Linking Reagents↗
PubMed1990

Red cell deformability and haematological disorders.

Blood rheology is the science of the flow and deformation of blood. Clinically, blood rheology is important because circulatory resistance has two major components, vascular and rheological. In large vessels, blood rheology should be considered in terms of bulk flow, the viscosity of blood depending mainly on red cell concentration and plasma viscosity and, to a lesser extent, on red cell deformability and aggregation. In the microcirculation, where cells must deform to pass through narrow capillaries, cellular rheology (i.e. the deformability of individual cells) is a major determinant of resistance to flow. This ability to deform is also a determinant of the cell's survival time in the circulation. The deformability of the red cell is essentially linked to its structure (i.e. its cellular geometry, membrane properties and cytoplasmic viscosity); thus structural abnormalities, as found in some haematological disorders, can be expected to affect blood flow in the microcirculation and/or red cell lifespan. Blood rheology is a relatively new discipline as applied to the practice of haematology. In 1985 the International Committee for Standardization in Haematology (ICSH) established an Expert Panel on Blood Rheology which has subsequently issued guidelines on the measurement of blood viscosity and erythrocyte deformability and on tests such as erythrocyte sedimentation rate and plasma viscosity that are used to monitor the acute phase response in inflammatory disease. Rheological methods now have sufficiently good sensitivity and specificity for their application to a wide variety of clinical disorders. This review illustrates their potential application to haematological disorders that cause abnormal deformability of red cells.

Erythrocyte Deformability↗
PubMed1990

Effects of deficiencies of glycophorins C and D on the physical properties of the red cell.

Red cells of the rare Leach phenotype lack the membrane glycophorins C and D, and a proportion of the red cells are elliptocytes. Judging from tests on suspensions of red cell ghosts sheared rotationally in an ektacytometer, it has previously been suggested that these membranes are relatively fragile and poorly deformable. We have carried out analyses of individual red cells to investigate possible factors which underlie the physical changes in these glycophorin-deficient cells. Micropipette analysis of the red cell membrane showed that the rigidity and viscosity were normal, both for elliptocytes and discocytes, for three donors deficient in glycophorins C and D. Red cell transit times through 5 microns pores, measured electronically for 2000 individual cells, showed no differences from controls. It was confirmed that the index of deformation obtained using an ektacytometer was reduced, but our results suggest that this arises from shape rather than membrane changes. The elliptocytes were found to have a lower volume and surface area than discocytes from the same donor (measured by micropipette aspiration of single red cells), and were rarely found in less dense red cell fractions. No reticulocytes were found to be elliptical. These data suggest that the elliptocytes are older red cells, and are formed from red cells which are initially released into the circulation with normal shape. Their elongated shape might arise from permanent distortion of the unstable membrane by shear forces in the circulation.

Elasticity↗
PubMed1990

Technological advances in blood rheology.

The science of blood rheology (study of the flow and deformability of blood) is of increasing practical importance to the investigation of blood disorders. In diagnostic laboratories, rheological tests such as the erythrocyte sedimentation rate, zeta sedimentation ratio, and plasma viscosity are used to monitor patients with an acute-phase response of greater than 24 h duration. In sickle-cell anemia, new methods for measuring erythrocyte deformability can be used to investigate the pathogenesis of vaso-occlusion, to test potential anti-sickling drugs, and to monitor drug efficacy in clinical trials. Genetic defects in the structure of the red cell membrane can have rheological consequences, monitoring of which may be useful for diagnosis. Rheological analysis of red cells infected by Plasmodium falciparum has indicated that their abnormal flow behavior may be an important pathological factor in malaria. Finally, the flow behavior of white blood cells, particularly neutrophils, is also important, as these cells, once activated, have the potential to occlude microvessels. The authors have reviewed the laboratory methodology and clinical applications that have led to recent advances in these aspects of blood rheology.

Acute-Phase Reaction↗
PubMed1990

Filterability of blood cells: methods and clinical applications.

It could be claimed that expansion in clinical aspects of haemorheology has largely been fueled by the development of a simple test of blood flow properties, i.e., analysis of filterability. With time, the level of sophistication in equipment, theory and sample preparation has increased. Theories for the development of flow in the filter have been described and these enable cellular parameters, such as transit times, to be calculated from experimental data. These theories can be quite general, and applied to filtration of red or white cells. Ideally, experimental design requires an understanding of cell behaviour at the filter and of the effects of factors such as the sample concentration and volume, and the presence of different types of cells or subpopulations. Otherwise, results are susceptible to misinterpretation, particularly if impurities or mixed populations of cells are present. It is thus very important to know accurately the constituents of the test suspension. In clinical applications the trend has been to move away from whole blood filtration, toward use of relatively pure suspension of separated red cells and white cells. In the area of red cell filtration this has led to reappraisal of some previously reported abnormalities. The relatively new study of white cell filtration should benefit from previous experience, but there is the added problem of the reactivity of the cells under test, and their mixed nature. In any case, critical evaluation of the meaning and clinical relevance of results is necessary.

Blood Flow Velocity↗
PubMed1989

Abnormalities in the mechanical properties of red blood cells caused by Plasmodium falciparum.

Although changes in the mechanical properties of infected red cells may contribute to the pathophysiology of malaria, such changes have not previously been described in detail. In this study, the physical properties of individual cells from both clinical and cultured samples infected with Plasmodium falciparum were tested using micropipette aspiration techniques. Cells containing ring forms took about 50% longer to enter 3 microns pipettes compared with nonparasitised cells, and there was a similar increase in the critical pressure required to induce cell entry. These abnormalities were similar in clinical and cultured samples. More mature cultured parasites (ie, trophozoites and schizonts containing pigment) caused much greater loss of deformability, with entry time and pressure increased four to sixfold. The decrease in deformability of the ring forms was attributable to a deficit in cell surface area/volume ratio (based on micropipette measurement of the surface area and volume of individual cells) and slight stiffening of the cell membrane (shear elastic modulus increased 13%, as measured by pipette aspiration of small membrane tongues). Measurement of the rate of cell shape recovery indicated that the membrane of parasitised cells was not more viscous. The main factor in the drastic loss of deformability of the trophozoites and schizonts was the presence of the large very resistant parasite itself. Otherwise, the cell surface area/volume deficit was slightly less and membrane rigidification slightly greater compared with ring forms. The above abnormalities should cause the trophozoites and schizonts to have great difficulty in traversing splenic or marrow sinuses and could contribute to microvascular occlusion and sequestration. On the other hand, the ring forms may be expected to circulate relatively unhindered.

Animals↗
PubMed1989

A study of red cell membrane properties in relation to malarial invasion.

The shape and mechanical properties of human red cells were modified in several ways and the consequences for the efficiency of invasion by Plasmodium falciparum in culture were investigated. Inhibition of invasion by depletion of ATP was shown to be unrelated to cell shape or deformability changes. Treatment of cells with N-ethylmaleimide (NEM), which dissociates some 70% of the native spectrin tetramers into the dimer, grossly reduced deformation of the cells under shear and increased by a factor of two or more the shear elastic modulus, as measured by the micropipette aspiration technique. Cells thus treated were efficiently invaded by P. falciparum (ca. 75% of control). In a population of cells pretreated with chlorpromazine, parasites were found in stomatocytic cells which were highly undeformable under shear. There was also considerable invasion into cells from subjects with hereditary pyropoikilocytosis, and two types of elliptocytosis. Cells treated with wheat germ agglutinin showed a dose-dependent increase in rigidity; a fivefold increase in elastic modulus (with total loss of deformation under shear in our conditions) still permitted invasion at a level of 50% of the control. The results suggest that gross mechanical properties of the membrane per se, at least within any physiologically relevant range, are unlikely to be the primary determinant of malarial invasion; this may instead be linked to the freedom of membrane proteins to migrate in the course of entry of the parasite.

2,3-Diphosphoglycerate↗
PubMed1989

Alteration of the mechanical properties of sickle cells by repetitive deoxygenation: role of calcium and the effects of calcium blockers.

The formation of dense, poorly deformable sickle cells was studied by subjecting pre-separated, less dense cells to repeated deoxygenation and reoxygenation for 15 h. In the presence of Ca (2 mmol/l), this process caused the number of irreversible sickled cells to increase five-fold, mean cell haemoglobin concentration to increase by 13% and cellular potassium to decrease by 22%. Also, red cell filterability through 5 microns filter pores was greatly worsened. These effects decreased but were not totally abolished when the extracellular Ca concentration was lowered to zero or 0.01 mmol/l. If a high K medium was used (135 mmol/l), cell swelling rather than shrinkage occurred. Swelling also occurred if ouabain was added to the incubation. The Ca-channel blockers nitrendipine and nisoldipine had different effects. Nitrendipine, in the range 10(-7) - 10(-5) mol/l, was partially protective against all the induced changes, but nisoldipine was not protective at 10(-8) or 10(-6) mol/l. Thus, deterioration in the properties of sickle cells appears to be linked to Ca-dependent potassium loss during repeated sickling and is inhibited by nitrendipine.

Anemia, Sickle Cell↗
PubMed1989

Changes in the flow properties of white blood cells after acute myocardial infarction.

Because they can obstruct blood vessels and release noxious substances, white blood cells may contribute to the development of tissue ischaemia. The flow properties of white cells were tested after myocardial infarction, by measuring the filtration rates of cell suspensions through 8 microns pore filters. Compared with mononuclear cells from age matched controls, mononuclear cells from patients with infarction showed impaired filterability within the first day after the onset of pain; this condition persisted for at least two days and by day 10 it was improved. On day 1, granulocyte filterability and the proportion showing morphological evidence of activation were nearly normal. By day 3 the flow resistance and activation had increased, but the changes seen depended on the age of the patient. The filterability and activation of granulocytes from patients aged less than 60 were significantly increased from day 1, whereas there were no changes in granulocytes from patients aged greater than 60 years. Suspensions of unfractionated white cells showed changes intermediate between the mononuclear cells and granulocytes. A group of five patients who presented with chest pain but who were subsequently found not to have had an infarction showed no evidence of abnormal filterability or activation. The changes in filterability probably reflect white cell activation, which may have an adverse effect on the perfusion of the ischaemic myocardium.

Adult↗
PubMed1989

Altered filtrability of white blood cells after myocardial infarction.

Abnormal white blood cell rheological behaviour has been implicated as a cause of blood flow disturbances under conditions of ischaemia and reduced perfusion pressure. Accordingly, we have tested the mechanical properties of white cells following myocardial infarction by measuring the rate at which suspension of these cells cause plugging of Nuclepore filters. The number of clogging particles in a standard white cell suspension increased by the third day after infarction but subsequently decreased to the control levels. Since white cells can cause blockage of narrow blood vessels, it is assumed that such changes in cellular properties may influence the eventual extent of infarction.

Aged↗
PubMed1988

White cell accumulation in dependent legs of patients with venous hypertension: a possible mechanism for trophic changes in the skin.

The mechanism by which chronic venous insufficiency and venous hypertension are associated with ulceration of the legs is not yet understood. To investigate this mechanism further accumulation of white cells in the dependent legs of normal volunteers, patients awaiting surgery for simple varicose veins, and patients with chronic venous insufficiency was studied. About 24% fewer white cells than in normal subjects left the dependent foot of patients with venous hypertension, and this trapping of white cells, was reversed when the foot was raised; similar changes were not observed in normal subjects or patients with varicose veins. The trophic skin changes typically seen in patients with venous hypertension may be aggravated by damage caused by the repeated accumulation of white cells in the microcirculation.

Adult↗