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Red Blood Cell Aggregation Characterization: Quantification and Modeling Implications of Red Blood Cell Aggregation at Low Shear RatesMehri, Rym January 2016 (has links)
Red blood cells (RBCs) are the most abundant cells in human blood, representing 40 to 45% of the blood volume (hematocrit). These cells have the particular ability to deform and bridge together to form aggregates under very low shear rates. The theory
and mechanics behind aggregation are, however, not yet completely understood.
The purpose of this work is to provide a novel method to analyze, understand and mimic blood behaviour in microcirculation. The main objective is to develop a methodology to quantify and characterize RBC aggregates and hence enhance the current understanding of the non-Newtonian behaviour of blood at the microscale. For this purpose, suspensions of porcine blood and human blood are tested in vitro in a Poly-di-methylsiloxane (PDMS) microchannel to characterize RBC aggregates within these two types of blood. These microchannels are fabricated using standard photolithography methods. Experiments are performed using a micro Particle Image Velocimetry ( PIV) system for shear rate measurements coupled with a high speed camera for the flow visualization.
Corresponding numerical simulations are conducted using a research Computational
Fluid Dynamic (CFD) solver, Nek5000, based on the spectral element method
solution to the incompressible non-Newtonian Navier-Stokes equations. RBC aggregate sizes are quantified in controlled and measurable shear rate environments for 5, 10 and 15% hematocrit. Aggregate sizes are determined using image processing techniques. Velocity fields of the blood flow are measured experimentally and compared to numerical simulations using simple non-Newtonian models (Power law and Carreau models).
This work establishes for the first time a relationship between RBC aggregate sizes
and corresponding shear rates in a microfluidic environment as well as one between RBC aggregate sizes and apparent blood viscosity at body temperature in a microfluidic controlled environment. The results of the investigation can be used to help develop new numerical models for non-Newtonian blood flow, provide a better understanding of the mechanics of RBC aggregation and help determine aggregate behaviour in clinical settings such as for degenerative diseases like diabetes and heart disease.
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Cellular lifespan based pharmacodynamic analysis of erythropoiesisFreise, Kevin Jay 01 July 2009 (has links)
The disposition of cells whose mechanism of death is related to the age of the cell cannot be appropriately represented by pharmacodynamic (PD) models where the elimination rate is related to the number of cells. In cells with age-related death their disposition is determined by their lifespan. Thus in these cells PD models of cellular response must incorporate a lifespan component. Previous cellular lifespan PD models assumed that the lifespan of cells is predetermined and does not vary over time. However, in many instances these assumptions are inappropriate and thus extensions to the existing models are needed. An important application of these time variant PD models is determining the erythropoiesis rate, since the lifespan of reticulocytes and mature erythrocytes are known to change over time under specific conditions.
The objectives us this work were to develop a general time variant lifespan-based PD model of cellular response and to use the model to determine the dynamic changes over time in both the erythrocyte lifespan and erythropoiesis rate under a variety of complex conditions. An initial time variant cellular lifespan model was formulated assuming no variability in lifespans and used to determine the dynamic changes in both the reticulocyte lifespan and erythropoiesis rate in sheep. Subsequently, the time variant model was extended to account for a distribution of cellular lifespans, which resulted in better capturing the physiology of sheep erythrocyte maturation. The model was then further extended to account for the effect of changes in the environment on cell lifespans and used to determine the effect of chemotherapy administration on sheep erythrocytes. In order to conduct studies on erythropoiesis in premature very low birth weight (VLBW) infants the ability to accurately measure erythrocytes and hemoglobin from clinically collected excess blood was validated. Then an in depth analysis of the relationship between erythropoietin, erythrocytes, and hemoglobin was conducted in a clinical study of premature VLBW infants that accounted for the dynamic hematological conditions experienced by these subjects. This analysis indicated that a nearly 4-fold increase in erythropoiesis could be achieved with only a modest increase in plasma erythropoietin concentrations.
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Blood Microflow Characterization Using Micro-Particle Image Velocimetry and 2-Beam Fluorescence Cross-Correlation SpectroscopyLe, Andy Vinh 04 December 2020 (has links)
Blood flow through microcirculation in both simple and complex geometry has been difficult to predict due to the composition and complex behavior of blood at the microscale. Blood is a dense suspension of deformable red blood cells that is comparable in dimensions to the microchannels that it flows through. As a result, rheological properties at the microscale can vastly differ from bulk rheological properties due to non-continuum effects. To further develop our understanding of blood microflow; experimental techniques should be explored.
In this work, we explore micro-particle image velocimetry (μPIV) and two-beam fluorescence cross-correlation spectroscopy (2bFCCS) in the application of characterizing blood in microflow conditions. For the development of the μPIV analysis, a polydimethylsiloxane co-flow channel is used to observe blood flow in controlled conditions. Flow conditions (velocity profile and blood layer thickness) are selected based on an analytical model and compared to experimental measurement. The experimental results presented indicate that current flow conditions are inadequate in providing a controlled rate of shear on the blood layer in the co-flow channel and further optimization are required to improve the measurement of the velocity profile. For the development of the 2bFCCS application for blood flow analysis, a wide glass capillary microfluidic device is used to complete the verification of fluorescence fluid admissibility, the effect of laser intensity on inducing photobleaching and the velocity measurement performance. The experimental measurement of the velocity profile is validated against the theoretical profile for a rectangular channel. Results of the velocity profile of high concentration red blood cells show promise in the technique’s ability to measure blood microflows closer to physiological conditions.
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Fragilidade osmótica eritrocitária e reticulocitometria em cães acometidos por doença renal crônica /Bizare, Amanda January 2020 (has links)
Orientador: Áureo Evangelista Santana / Resumo: A anemia é considerada um dos fatores para avaliar progressão da doença renal e diminuição da qualidade de vida do paciente. Conforme a doença renal progride, ocorre aumento gradativo na produção de toxinas urêmicas que reduz a meia vida dos eritrócitos circulantes por interferir na estabilidade da membrana eritrocitária. Para tanto, utiliza-se a contagem de reticulócitos para classificar a anemia como regenerativa ou não regenerativa. Objetivou-se neste estudo avaliar a resistência da membrana das hemácias, utilizando-se do teste de Fragilidade Osmótica Eritrocitária (FOE) em cães com doença renal crônica (DRC) e avaliação de reticulócitos. Foram avaliados 43 cães provenientes da rotina do Serviço de Nefrologia e Urologia do Hospital Veterinário Governador Laudo Natel da Faculdade de Ciências Agrárias e Veterinárias - UNESP - campus Jaboticabal. As referidas unidades experimentais foram distribuídas em três grupos, quais sejam, G0 (n=13), composto por cães hígidos e G1, DRC estádios 1 e 2 (n=14) e G2, DRC estádios 3 e 4 (n=16), classificados de acordo com o recomendado pela International Renal Interest Society. A fim de definir os critérios de inclusão dos cães foram feitos, além do exame físico, a avaliação de pressão arterial, hemograma, contagem de reticulócitos, exames bioquímicos, urinálise e relação proteína/creatinina urinária (UP/C). Para execução do teste de FOE as hemácias foram diluídas em concentrações decrescentes de cloreto de sódio e analisadas por citometria ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Anemia is considered one of the factors to assess the kidney disease progress and the decrease in patient's quality of life. As kidney disease progresses, there is a gradual increase in urinary toxin production that shortens the circulating erythrocyte half-life by interfering with erythrocyte membrane stability. To do this, use a reticulocyte count to classify anemia as regenerative or non-regenerative. The objectives of this study were to evaluate the resistance of the red blood cells, using the Erythrocyte Osmotic Fragility test in dogs with chronic kidney disease (CKD) and to evaluate reticulocytes. Forty-three dogs were charged, followed by the routine of the Nephrology and Urology Service of the Governor Laudo Natel Veterinary Hospital of the Faculty of Agricultural and Veterinary Sciences - UNESP - Campus Jaboticabal. The experimental units were divided into three groups, namely, G0 (n = 13), consisting of healthy dogs and G1, CKD stages 1 and 2 (n = 14) and G2, CKD stages 3 and 4 (n = 16), classification proposed by the International Renal Interest Society. In order to define the inclusion criteria of dogs made, in addition to physical examination, an assessment of blood pressure, blood count, reticulocyte count, biochemical tests, urinalysis, and urinary protein/creatinine ratio. To perform the erythrocyte osmotic fragility test, red blood cells were diluted in decreasing sodium chloride filters (0.9 to 0.0%) and analyzed by flow cytometry. As creatinine serum concen... (Complete abstract click electronic access below) / Mestre
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Decreased Total Carbonic Anhydrase Esterase Activity and Decreased Levels of Carbonic Anhydrase 1 Isozyme in Erythrocytes of Type II Diabetic PatientsGambhir, Kanwal K., Ornasir, Jehan, Headings, Verle, Bonar, Adolphus 01 June 2007 (has links)
In this exploratory study, we investigated total erythrocyte carbonic anhydrase (CA) estrase activity as well as CA I isozyme concentration in patients with diabetes mellitus type II (DM) and healthy individuals of Howard University Hospital community. Total estrase activity of CA was measured spectrophotometrically using p-nitrophenol acetate before and after inhibition with acetazolamide. CA I isozyme was measured by radial immunodiffusion using monoclonal antibody (CA I) in agarose plates. The study involved 20 consented participants; 10 normal (N) and 10 (DM), 21 to 84 years of age. The study was approved by the Howard University Institution Review Board. The CA activity was measured following lysis of cells as U/min/mL and CA I concentration as mg/l. We observed CA activity as 46.3±4(N) and 25±2.1 (DM) whereas CA I concentration as 1896±125 (N) and 1104 ±63 (DM). We speculate that the change in the CA activity may of fundamental importance in the regulation of intracellular; pHi for the basic control of metabolism in diabetes mellitus. Further, we propose that CA activity is a good candidate for a biomarker of diabetes mellitus for the early detection of insulin resistance because the CA activity variation was proportional to the severity of the diabetes.
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Biophysics of Blood MembranesHimbert, Sebastian 11 1900 (has links)
Red blood cells (RBCs) are the predominant cell type in blood and have a two-layered outer shell which is composed of a cytoskeleton network tethered to a cytoplasmic membrane.
In this thesis, I study the structure and mechanical properties of the RBC’s cytoplasmic membrane (RBCcm) on the nanoscale and utilize this knowledge to functionalize this biological structure on a molecular level. In a first case study, I measure the membrane’s bending rigidity from thermal fluctuations observed in X-ray diffuse scattering (XDS) and Neutron Spin Echo (NSE) experiments, as well as Molecular Dynamics (MD) simulations. I provide evidence of the RBCcm's highly deformable nature with a bending rigidity that is substantially softer as compared to synthetic membranes. The methods are applied to RBCs that were stored for up to 5 weeks. I demonstrate that storage of RBCs leads to an increased fraction of liquid ordered membrane domains and an increased bending rigidity.
RBCs are ideal for pharmaceutical applications as they provide access to numerous targets in the body, however lack specificity. Functionalizing the cytoplasmic membrane is thus a prerequisite to use these cells in biotechnology. I develop protocols throughout two studies to tune the membrane's lipid and protein composition. I investigate the impact of synthetic lipid molecules on the membrane's structure and demonstrate that small molecules can be encapsulated into liposomes that are formed from these hybrid membranes.
Further, I provide direct evidence that the SARS-CoV 2 spike protein can be anchored into the RBCcm through a detergent mediated insertion protocol. These virus-like particles are observed to trigger seroconversion in mouse models, which demonstrates the potential of functionalized RBC in biotechnology. / Thesis / Doctor of Philosophy (PhD)
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The Effects of the Endothelial Surface Layer on Red Blood Cell Dynamics in Microvessel BifurcationsCarlson Bernard Triebold (11198889) 28 July 2021 (has links)
<div>Red blood cells (RBCs) make up 40-45% of blood and play an important role in oxygen transport. That transport depends on the RBC distribution throughout the body, which is highly heterogeneous. That distribution, in turn, depends on how RBCs are distributed or partitioned at diverging vessel bifurcations where one vessel flows into two. Several studies have used mathematical modeling to consider RBC partitioning at such bifurcations in order to produce useful insights. However, these studies assume that the vessel wall is a flat impenetrable homogeneous surface. While this is a good first approximation, especially for larger vessels, the vessel wall is typically coated by a flexible, porous endothelial surface layer (ESL) that is 0.5-1 microns thick. To better understand the possible effects of this layer on RBC partitioning, a diverging capillary bifurcation is analyzed using a flexible, two-dimensional RBC model. The model is also used to investigate RBC deformation and penetration of the ESL region when ESL properties are varied. The RBC is represented using interconnected viscoelastic elements. Stokes flow equations (viscous flow) model the surrounding fluid. The flow in the ESL is modeled using the Brinkman approximation for porous media with a corresponding hydraulic resistivity. The resistance of the ESL to compression is modeled using an osmotic pressure difference. The study includes isolated cells that pass through the bifurcation one at a time with no cell-cell interactions and two cells that pass through the bifurcation at the same time and interact with each other. A range of physiologically relevant hydraulic resistivities and osmotic pressure differences are explored.</div><div><br></div><div>For isolated cell simulations, decreasing hydraulic resistivity and/or decreasing osmotic pressure difference produced four behaviors: 1) RBC distribution nonuniformity increased; 2) RBC deformation decreased; 3) RBCs slowed down slightly; and 4) RBCs penetrated more deeply into the ESL. The presence of an altered flow profile and the ESL's resistance to penetration were primary factors responsible for these behaviors. In certain scenarios, ESL penetration was deep enough to present a possibility of cell adhesion, as can occur in pathological situations.</div><div><br></div><div>For paired cell simulations, more significant and complex changes were observed. Three types of effects that alter partitioning as hydraulic resistivity is changed are identified. Decreasing hydraulic resistivity in the ESL produced lower RBC deformation. Including cell-cell interactions tended to increase deformation sharply compared to isolated cell scenarios. ESL penetration generally decreased for lower hydraulic resistivities except in scenarios with significant cell-cell interactions. This was primarily due to changes in flow profiles induced by the altered hydraulic resistivity levels.</div>
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Experimental and Computational Modeling of Ultrasound Correlation TechniquesGeorge, Brian Patrick 19 May 2010 (has links)
No description available.
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Vesiculation of Red Blood Cells in the Blood Bank: A Multi-Omics Approach towards Identification of Causes and ConsequencesFreitas Leal, Joames F., Lasonder, Edwin, Sharma, Vikram, Schiller, Jürgen, Fanelli, Giuseppina, Rinalducci, Sara, Brock, Roland, Bosman, Giel 19 April 2023 (has links)
Microvesicle generation is an integral part of the aging process of red blood cells in vivo and in vitro. Extensive vesiculation impairs function and survival of red blood cells after transfusion, and microvesicles contribute to transfusion reactions. The triggers and mechanisms of microvesicle generation are largely unknown. In this study, we combined morphological, immunochemical, proteomic, lipidomic, and metabolomic analyses to obtain an integrated understanding of the mechanisms underlying microvesicle generation during the storage of red blood cell concentrates. Our data indicate that changes in membrane organization, triggered by altered protein conformation, constitute the main mechanism of vesiculation, and precede changes in lipid organization. The resulting selective accumulation of membrane components in microvesicles is accompanied by the recruitment of plasma proteins involved in inflammation and coagulation. Our data may serve as a basis for further dissection of the fundamental mechanisms of red blood cell aging and vesiculation, for identifying the cause-effect relationship between blood bank storage and transfusion complications, and for assessing the role of microvesicles in pathologies affecting red blood cells.
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Computational Modeling of Intracapillary Bacteria Transport in Tumor MicrovasculatureWindes, Peter 06 October 2016 (has links)
The delivery of drugs into solid tumors is not trivial due to obstructions in the tumor microenvironment. Innovative drug delivery vehicles are currently being designed to overcome this challenge. In this research, computational fluid dynamics (CFD) simulations were used to evaluate the behavior of several drug delivery vectors in tumor capillaries—specifically motile bacteria, non-motile bacteria, and nanoparticles. Red blood cells, bacteria, and nanoparticles were imposed in the flow using the immersed boundary method. A human capillary model was developed using a novel method of handling deformable red blood cells (RBC). The capillary model was validated with experimental data from the literature. A stochastic model of bacteria motility was defined based on experimentally observed run and tumble behavior. The capillary and bacteria models were combined to simulate the intracapillary transport of bacteria. Non-motile bacteria and nanoparticles of 200 nm, 300 nm, and 405 nm were also simulated in capillary flow for comparison to motile bacteria. Motile bacteria tended to swim into the plasma layer near the capillary wall, while non-motile bacteria tended to get caught in the bolus flow between the RBCs. The nanoparticles were more impacted by Brownian motion and small scale fluid fluctuations, so they did not trend toward a single region of the flow. Motile bacteria were found to have the longest residence time in a 1 mm long capillary as well as the highest average radial velocity. This suggests motile bacteria may enter the interstitium at a higher rate than non-motile bacteria or nanoparticles of diameters between 200–405 nm. / Master of Science
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