Morphometry and Stiffness of Red Blood Cells—Signatures of Neurodegenerative Diseases and Aging

Human red blood cells (RBCs) are unique cells with the remarkable ability to deform, which is crucial for their oxygen transport function, and which can be significantly altered under pathophysiological conditions. Here we performed ultrastructural analysis of RBCs as a peripheral cell model, looking for specific signatures of the neurodegenerative pathologies (NDDs)—Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS) and Alzheimer’s disease (AD), utilizing atomic force (AFM) and conventional optical (OM) microscopy. We found significant differences in the morphology and stiffness of RBCs isolated from patients with the selected NDDs and those from healthy individuals. Neurodegenerative pathologies' RBCs are characterized by a reduced abundance of biconcave discoid shape, lower surface roughness and a higher Young’s modulus, compared to healthy cells. Although reduced, the biconcave is still the predominant shape in ALS and AD cells, while the morphology of PD is dominated by crenate cells. The features of RBCs underwent a marked aging-induced transformation, which followed different aging pathways for NDDs and normal healthy states. It was found that the diameter, height and volume of the different cell shape types have different values for NDDs and healthy cells. Common and specific morphological signatures of the NDDs were identified.

Download Full-text

Passive haemagglutination test for human neurocysticercosis immunodiagnosis: II - Comparison of two standardized procedures for the passive haemagglutination reagent in the detection of anti-Cysticercus cellulosae antibodies in cerebrospinal fluids

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46651988000100010 ◽

1988 ◽

Vol 30 (1) ◽

pp. 57-62 ◽

Cited By ~ 2

Author(s):

Mirthes Ueda ◽

Adelaide José Vaz ◽

Eide Dias Camargo ◽

Ana Maria Carvalho de Souza ◽

Regina Maria Figueiredo Benelli ◽

...

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Treated Group ◽

Healthy Individuals ◽

Saline Extract ◽

Passive Haemagglutination ◽

Haemagglutination Test ◽

Human Red Blood Cells ◽

Cysticercus Cellulosae ◽

Level Of Significance

A comparison of two different standardized reagent procedures for the passive haemagglutination test (PHA) in the detection of specific antibody to Cysticercus cellulosae in cerebrospinal fluid (CSF) was carried out. The formaldehyde-treated group O Rh-human red blood cells (HuRBC) and glutaraldehyde-treated sheep red blood cells (SRBC) were the supplies for the reagents preparation and, in the tests, they were designated as PHA-1 and PHA-2, respectively. For both reagents the cells were coated with the cysticerci total saline extract (TS) antigen. PHA-1 and PHA-2 were assessed in a total of 204 CSF from patients with neurocysticercosis, from non-related infections and from healthy individuals. The positivity and specificity indices obtained were respectively 81.7% and 94.4% for PHA-1 and for PHA-2, 88.7% and 96.6%. Since no significant differences were observed between the results provided by two reagents, at level of significance of 0.05, either processes of cell sensitization can alternatively be used according to the own laboratory convenience.

Download Full-text

Coronavirus SARS-CoV-2: Hypotheses of Impact on the Circulatory System, Prospects for the Use of Perfluorocarbon Emulsion, and Feasibility of Biophysical Research Methods

General Reanimatology ◽

10.15360/1813-9779-2020-3-0-1 ◽

2020 ◽

Vol 16 (3) ◽

pp. 4-13 ◽

Cited By ~ 1

Author(s):

V. V. Moroz ◽

A. M. Chernysh ◽

Elena K. Kozlova

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Molecular Mechanisms ◽

Circulatory System ◽

High Resolution Spectroscopy ◽

Perfluorocarbon Emulsion ◽

Erythrocyte Morphology ◽

Force Microscopy ◽

Human Red Blood Cells ◽

Atomic Force

This paper highlights published hypotheses on the possibility of coronavirus SARS-CoV-2 entry into the bloodstream, its interaction with vascular endothelium, red blood cells, hemoglobin and its fragments. As a result of such interaction, iron ions may be released into the bloodstream and, subsequently, a cytokine storm may occur. In this context, it is important to find a cytoprotective agent capable of blocking such processes. The perfluorocarbon emulsion could be a candidate for this role.The aim of the paper is to show the feasibility of biophysical methods to study the molecular mechanisms of action of SARS-CoV-2 on human red blood cells and hemoglobin as well as the restorative and cytoprotective effect of the perfluorocarbon emulsion during Fe2+ oxidation in heme.Materials and methods. High resolution spectroscopy, atomic force microscopy, atomic force spectroscopy, electroporation were used. Blood was exposed to oxidizing agents of different nature. Perfluorocarbon emulsion was added in various concentrations and its effect at various incubation times was studied. Concentration of hemoglobin derivatives was calculated considering multicollinearity, and statistical analysis of the results was performed.Results. The perfluorocarbon emulsion was shown to have an effective restorative and cytoprotective action in iron ion oxidation in the heme: Fe3+ was restored to Fe2+. The degree of MetHb reduction to HbO2 and Hb depended on the concentration of the oxidizing agent and incubation time. We observed a change in MetHb content from 80-90% to 5-12%. The perfluorocarbon emulsion in clinical concentrations helped eliminate local membrane defects and restored normal erythrocyte morphology.Conclusion. In the light of the studied hypotheses, the use of perfluorocarbon emulsion can become an effective method for blocking the consequences of coronavirus effect on the blood cells and restoring a normal gas exchange.

Download Full-text

Desaggregation of Human Red Blood Cells by Various Surface-Active Agents as Related to Changes of Cell Shape and Hemolysis

Acta Haematologica ◽

10.1159/000208160 ◽

1975 ◽

Vol 53 (2) ◽

pp. 82-89 ◽

Cited By ~ 3

Author(s):

P. Gaehtgens ◽

K.U. Benner

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Cell Shape ◽

Human Red Blood Cells ◽

Surface Active Agents ◽

Surface Active

Download Full-text

Measurements of Mechanical Properties of Human Red Blood Cells

Microelectromechanical Systems ◽

10.1115/imece2006-15175 ◽

2006 ◽

Author(s):

Choongbae Park ◽

Steven T. Wereley ◽

Osvaldo H. Campanella ◽

David E. Nivens ◽

Kenneth M. Little ◽

...

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Thermal Stresses ◽

Phase Lag ◽

Human Red Blood Cells ◽

Atomic Force ◽

Spherical Bead ◽

Novel Method ◽

Height Change ◽

Afm Cantilever

We developed a novel method to measure the rheological properties of single red blood cells (RBC) using the atomic force microscope (AFM). A spherical bead at the AFM cantilever tip compressed and relaxed the RBC. The force and displacement were converted into effective stress and strain. The impulse viscoelastic technique was used to compute the effective storage (E') and loss (E") moduli and phase lag (δ). Unfixed and fixed red blood cells were tested. Both cells were on glass coated with poly-l-lysine and then kept in phosphate buffered saline (PBS) until the experiment was finished. Measurements were done with height change and force up to 451nm and 64nN. The cells were found to be quite elastic, with phase lag on the order of 10-2 to 10-1 rad. Stepped changes in oscillation rate from 0.5Hz to 2.5Hz did not result in significant change in the measured results. To improve accuracy, we also design a bimaterial cantilever which consists of a gold layer on silicon with controlled thermal stresses such that the cantilever is curved. The curvature allows the root to fits the angle of the AFM head and the tip to be parallel to the substrate so that the RBC is squeezed between two parallel surfaces.

Download Full-text

Quantitative Scanning Condition Effects on Atomic Force Microscopy Measurements of Human Red Blood Cells

Advanced Science Letters ◽

10.1166/asl.2011.1880 ◽

2011 ◽

Vol 4 (11) ◽

pp. 3637-3643 ◽

Cited By ~ 1

Author(s):

R. C. Curley ◽

L. P. Silva

Keyword(s):

Atomic Force Microscopy ◽

Red Blood Cells ◽

Blood Cells ◽

Force Microscopy ◽

Human Red Blood Cells ◽

Atomic Force

Download Full-text

Relating the blood-thinning effect of pentoxifylline to the reduction in the elastic modulus of human red blood cells: anin vivostudy

Biomaterials Science ◽

10.1039/c8bm01691g ◽

2019 ◽

Vol 7 (6) ◽

pp. 2545-2551 ◽

Cited By ~ 3

Author(s):

Katerina E. Aifantis ◽

Sanjiv Shrivastava ◽

Sygkliti-Henrietta Pelidou ◽

Alfonso H. W. Ngan ◽

Stavros I. Baloyannis

Keyword(s):

Elastic Modulus ◽

Red Blood Cells ◽

Blood Cells ◽

Human Subjects ◽

Healthy Human ◽

Force Microscopy ◽

Human Red Blood Cells ◽

Atomic Force ◽

Healthy Human Subjects ◽

Thinning Effect

Atomic force microscopy indentation was employed to illustrate that pentoxifylline reduces the elastic modulus of red blood cells (from healthy human subjects), increasing hence their flexibility.

Download Full-text

Changes of cell shape and surface charge topography in ATP-depleted human red blood cells

Mechanisms of Ageing and Development ◽

10.1016/0047-6374(85)90070-3 ◽

1985 ◽

Vol 29 (3) ◽

pp. 309-316 ◽

Cited By ~ 15

Author(s):

Yehuda Marikovsky ◽

Ronald S. Weinstein ◽

Ehud Skutelsky ◽

David Danon

Keyword(s):

Red Blood Cells ◽

Surface Charge ◽

Blood Cells ◽

Cell Shape ◽

Human Red Blood Cells

Download Full-text

The circulatory physiopathology of human red blood cells investigated with a multiplatform model of cellular homeostasis. II. Capillary transits and the role of PIEZO1

10.1101/2020.03.07.981787 ◽

2020 ◽

Cited By ~ 2

Author(s):

Simon Rogers ◽

Virgilio L. Lew

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Time Course ◽

Capillary Flow ◽

Cell Model ◽

Recovery Period ◽

Large Body ◽

Human Red Blood Cells ◽

Early Results ◽

Depth Study

AbstractIn this and the next paper of this series we apply the red cell model introduced in the previous paper to investigate the changes in RBC homeostasis during capillary transits and over the full circulatory lifespan of the cells. These are topics inaccessible to direct experimentation but rendered mature for a modelling approach by recent findings and by a large body of apparently unrelated early results which robustly constrain the parameter space offering the opportunity for an in depth study of the mechanisms involved. Capillary transit times vary between 0.5 and 1.5s during which the red blood cells squeeze and deform in the capillary stream transiently opening stress-gated PIEZO1 channels, creating minuscule quantal changes in RBC ion contents and volume. Widely accepted early views originally based on results from experimentally shear-stressed red cells suggested that quantal changes generated during capillary transits add up over time to generate the documented changes in RBC density during their long circulatory lifespan, the quantal hypothesis. Applying the new PIEZO1 extension of the RBC model (RCM) introduced in the previous paper we investigated in detail the changes in homeostatic variables that may be expected during single capillary transits resulting from transient PIEZO1 channel activation. The predicted quantal volume changes were infinitesimal in magnitude, biphasic in nature, and essentially irreversible within inter-transit periods. A sub-second transient PIEZO1 activation triggered a sharp swelling peak followed by a much slower recovery period towards lower-than-baseline volumes. The peak response was caused by net CaCl2 and fluid gain via PIEZO1 channels driven by the steep electrochemical inward Ca2+ gradient. The ensuing dehydration followed a complex time-course with sequential, but partially overlapping contributions by KCl loss via Ca2+-activated Gardos channels, PMCA mediated calcium extrusion and chloride efflux by the Jacobs-Steward mechanism. The change in relative cell volume predicted for single capillary transits was below 10−4, an infinitesimal volume change incompatible with a functional role in capillary flow. The biphasic response predicted by the RCM appears to conform to the quantal hypothesis, but whether its cumulative effects could account for the documented changes in density during RBC senescence required an investigation of the effects of myriad transits over the full period of circulatory lifespan, the subject of the next paper of this series.

Download Full-text

Up-down biphasic volume response of human red blood cells to PIEZO1 activation during capillary transits

PLoS Computational Biology ◽

10.1371/journal.pcbi.1008706 ◽

2021 ◽

Vol 17 (3) ◽

pp. e1008706 ◽

Cited By ~ 2

Author(s):

Simon Rogers ◽

Virgilio L. Lew

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Time Course ◽

Capillary Flow ◽

Cell Model ◽

Recovery Period ◽

Large Body ◽

Computational Modelling ◽

Companion Paper ◽

Human Red Blood Cells

In this paper we apply a novel JAVA version of a model on the homeostasis of human red blood cells (RBCs) to investigate the changes RBCs experience during single capillary transits. In the companion paper we apply a model extension to investigate the changes in RBC homeostasis over the approximately 200000 capillary transits during the ~120 days lifespan of the cells. These are topics inaccessible to direct experimentation but rendered mature for a computational modelling approach by the large body of recent and early experimental results which robustly constrain the range of parameter values and model outcomes, offering a unique opportunity for an in depth study of the mechanisms involved. Capillary transit times vary between 0.5 and 1.5s during which the red blood cells squeeze and deform in the capillary stream transiently opening stress-gated PIEZO1 channels allowing ion gradient dissipation and creating minuscule quantal changes in RBC ion contents and volume. Widely accepted views, based on the effects of experimental shear stress on human RBCs, suggested that quantal changes generated during capillary transits add up over time to develop the documented changes in RBC density and composition during their long circulatory lifespan, the quantal hypothesis. Applying the new red cell model (RCM) we investigated here the changes in homeostatic variables that may be expected during single capillary transits resulting from transient PIEZO1 channel activation. The predicted quantal volume changes were infinitesimal in magnitude, biphasic in nature, and essentially irreversible within inter-transit periods. A sub-second transient PIEZO1 activation triggered a sharp swelling peak followed by a much slower recovery period towards lower-than-baseline volumes. The peak response was caused by net CaCl2 and fluid gain via PIEZO1 channels driven by the steep electrochemical inward Ca2+ gradient. The ensuing dehydration followed a complex time-course with sequential, but partially overlapping contributions by KCl loss via Ca2+-activated Gardos channels, restorative Ca2+ extrusion by the plasma membrane calcium pump, and chloride efflux by the Jacobs-Steward mechanism. The change in relative cell volume predicted for single capillary transits was around 10−5, an infinitesimal volume change incompatible with a functional role in capillary flow. The biphasic response predicted by the RCM appears to conform to the quantal hypothesis, but whether its cumulative effects could account for the documented changes in density during RBC senescence required an investigation of the effects of myriad transits over the full four months circulatory lifespan of the cells, the subject of the next paper.

Download Full-text