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Microfluidic devices are frequently manufactured with polydimethylsiloxane (PDMS) due to its affordability, transparency, and simplicity. However, high-pressure flow through PDMS microfluidic channels lead to an increase in channel size due to the compliance of the material. As a result, longer response times are required to reach steady flow rates, which increases the overall time required to complete experiments when using a syringe pump. Due to its excellent optical properties and increased rigidity, Norland Optical Adhesive (NOA) has been proposed as a promising material candidate for microfluidic fabrication. This study compares the compliance and deformation properties of three different characteristic sized (width of parallel channels: 100, 40 and 20 µm) microfluidic devices made of PDMS and NOA. The comparison of the microfluidics devices is made based on the Young's modulus, roughness, contact angle, channel width deformation, flow resistance and compliance. The experimental resistance is estimated through the measurement of the flow at a given pressure and a precision flow meter. The characteristic time of the system is extracted by fitting the two-element resistance-compliance (RC) hydraulic circuit model. The compliance of the microfluidics chips is estimated through the measurement of the characteristic time required for channels to achieve an output flow rate equivalent to that of the input flow rate using a syringe pump and a precision flow meter. The Young modulus was found to be 2 MPa for the PDMS and 1743 MPa for the NOA 63. The surface roughness was found to be higher for the NOA 63 than for the PDMS. The hydrophilicities of materials were found comparable with and without plasma treatment. The results show that NOA devices have lower compliance and deformation than PDMS devices.
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This method describes the use of thick round borosilicate glass micro-channels for blood flow visualization using micro-particle image velocimetry (µPIV) techniques. In contrast with popular methods using squared polydimethylsiloxane channels, this method allows for visualization of blood flow in channel geometries that resemble more the natural physiology of human blood vessels. With a custom designed enclosure, the microchannels were submerged in glycerol to reduce light refraction occurring during µPIV due to the thick walls of the glass channels. A method is proposed to correct the extracted velocity profiles from the µPIV to account for out-of-focus error. The customized elements of this method include: ⢠The use of thick circular glass micro-channels, ⢠a custom designed mounting solution for the channels on a glass slide for flow visualization, ⢠a MATLAB code to correct velocity profile accounting for out-of-focus error.
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Patients with sickle cell disease (SCD) have poorly deformable red blood cells (RBC) that may impede blood flow into microcirculation. Very few studies have been able to directly visualize microcirculation in humans with SCD. Sublingual video microscopy was performed in eight healthy (HbAA genotype) and four sickle cell individuals (HbSS genotype). Their hematocrit, blood viscosity, red blood cell deformability, and aggregation were individually determined through blood sample collections. Their microcirculation morphology (vessel density and diameter) and microcirculation hemodynamics (local velocity, local viscosity, and local red blood cell deformability) were investigated. The De Backer score was higher (15.9 mm-1) in HbSS individuals compared to HbAA individuals (11.1 mm-1). RBC deformability, derived from their local hemodynamic condition, was lower in HbSS individuals compared to HbAA individuals for vessels < 20 µm. Despite the presence of more rigid RBCs in HbSS individuals, their lower hematocrit caused their viscosity to be lower in microcirculation compared to that of HbAA individuals. The shear stress for all the vessel diameters was not different between HbSS and HbAA individuals. The local velocity and shear rates tended to be higher in HbSS individuals than in HbAA individuals, notably so in the smallest vessels, which could limit RBC entrapment into microcirculation. Our study offered a novel approach to studying the pathophysiological mechanisms of SCD with new biological/physiological markers that could be useful for characterizing the disease activity.
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Anemia Falciforme , Soalho Bucal , Humanos , Microcirculação/fisiologia , Hemodinâmica , Eritrócitos/fisiologia , Hemoglobina Falciforme , ReologiaRESUMO
In microcirculation, red blood cells (RBCs) tend to migrate toward the centre of the vessel leaving a region of a cell depleted layer or cell-free layer (CFL) at the vessel wall and a core of RBCs at the centre. This heterogenous distribution of cells has an effect on the blood apparent viscosity and the exchanges of gases and nutrients between the RBCs and the vessel. Understanding the formation of the CFL and obtaining accurate measurement of it is paramount for furthering development of devices such as drug administration. This paper presents a general semi-automatic method to quantify the thickness of the CFL for different channel geometries and image quality. It enables the use of a method based on intensity, a method using the gradient of the intensity, or a method based on spatiotemporal variation. The main features are reported, the performance is demonstrated on experimentally obtained image sets and accuracy is validated using synthetic images with known CFL thickness. A pure automatic detection is limited by the most visually correct using the spatiotemporal method, however proposed thresholding through automatic detection allows for quality controls through manual adjustments. With a semi-automatic approach RBC core variability between 3 % to 8 % was found when the test user was tasked with repeating the analysis of the same set. The presented method provides, users without programming ability with a user-friendly interface that can extract CFL automatically with quality control through manual adjustments.
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Eritrócitos , Hemodinâmica , Viscosidade Sanguínea , MicrocirculaçãoRESUMO
Teacher registration is increasingly utilised as a governance mechanism to audit teachers' work and drive professional practice. There is limited and mixed empirical evidence, however, as to whether registration drives teaching quality. Our study extends this limited empirical base by critically examining the policy trajectory in Australia to bring early childhood teachers into a uniform system of registration with primary and secondary teachers. Adopting a relatively novel methodology, the study intertwined a critical social policy framing with a national quantitative survey. Results showed that respondents perceived their professional self, followed by their workplace (colleagues and employer) as key influencers of quality practice, and neither agreed nor disagreed that teacher registration was beneficial. Findings problematise the need for, and benefits of, teacher registration. That early childhood teachers' practice and development was most driven by intrinsic motivation and, to a lesser extent, being employed in high-quality, not-for-profit, and preschool settings where other early childhood teachers are employed, suggests that more effective and progressive policy approaches to support quality early childhood education require an addressing of the contexts and conditions in which early childhood teachers work.
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Predicting blood microflow in both simple and complex geometries is challenging because of the composition and behavior of the blood at microscale. However, characterization of the velocity in microchannels is the key for gaining insights into cellular interactions at the microscale, mechanisms of diseases, and efficacy of therapeutic solutions. Image-based measurement techniques are a subset of methods for measuring the local flow velocity that typically utilize tracer particles for flow visualization. In the most basic form, a high-speed camera and microscope setup are the only requirements for data acquisition; however, the development of image processing algorithms and equipment has made current image-based techniques more sophisticated. This mini review aims to provide a succinct and accessible overview of image-based experimental measurement techniques to characterize the velocity field of blood microflow. The following techniques are introduced: cell tracking velocimetry, kymographs, micro-particle velocimetry, and dual-slit photometry as entry techniques for measuring various velocity fields either in vivo or in vitro.
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Although quantitative ultrasound techniques based on the parameterization of the backscatter coefficient (BSC) have been successfully applied to blood characterization, theoretical scattering models assume blood as an isotropic scattering medium. However, the red blood cell (RBC) aggregates form anisotropic structures such as rouleaux. The present study proposes an anisotropic formulation of the effective medium theory combined with the local monodisperse approximation (EMTLMA) that considers perfectly aligned prolate-shaped aggregates. Theoretical BSC predictions were first compared with computer simulations of BSCs in a forward problem framework. Computer simulations were conducted for perfectly aligned prolate-shaped aggregates and more complex configurations with partially aligned prolate-shaped aggregates for which the size and orientation of RBC aggregates were obtained from blood optical observations. The isotropic and anisotropic EMTLMA models were then compared in an inverse problem framework to estimate blindly the structural parameters of RBC aggregates from the simulated BSCs. When considering the isotropic EMTLMA, the use of averaged BSCs over different insonification directions significantly improves the estimation of aggregate structural parameters. Overall, the anisotropic EMTLMA was found to be superior to the isotropic EMTLMA in estimating the scatterer volume distribution. These results contribute to a better interpretation of scatterer size estimates for blood characterization.
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Eritrócitos , Ultrassom , Anisotropia , Simulação por Computador , UltrassonografiaRESUMO
Blood vessels in living tissues are an organized and hierarchical network of arteries, arterioles, capillaries, veinules and veins. Their sizes, lengths, shapes and connectivity are set up for an optimum perfusion of the tissues in which they deploy. In order to study the hemodynamics and hemophysics of blood flows and also to investigate artificial vasculature for organs on a chip, it is essential to reproduce most of these geometric features. Common microfluidic techniques produce channels with a uniform height and a rectangular cross section that do not capture the size hierarchy observed in vivo. This paper presents a new single-mask photolithography process using an optical diffuser to produce a backside exposure leading to microchannels with both a rounded cross section and a direct proportionality between local height and local width, allowing a one-step design of intrinsically hierarchical networks.
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Velocidade do Fluxo Sanguíneo , Técnicas Analíticas Microfluídicas , Humanos , Tamanho da PartículaRESUMO
The wall adhesion of 1⯵m microparticles in human blood was studied in circular microchannels. The level of particle wall adhesion was measured for varying levels of shear rate and varying degrees of red blood cell aggregation, which was modulated by the addition of macromolecule dextran 500. The blood preparations were injected into PDMS microfluidic devices that were modified to have circular channels, better matching the geometry of physiological microcirculation compared to square channels or Couette flow systems. The circular walls of the microchannels were embedded with biotinylated phospholipids to which marginating microspheres coated with streptavidin bound. The particle wall adhesion was evaluated by counting the particles adhering to the channel wall after flushing the channel. Blood preparations of five dextran concentrations (including baseline case of 0%) were tested for four flow velocities, to quantify the effects of aggregation for varying shear rate. It was found that the level of particle wall adhesion was positively correlated with the level of RBC aggregation, particularly at low shear rates, when aggregation was enhanced. The particle adhesion was especially enhanceat aggregation levels in the range of physiological aggregation levels of whole blood, suggesting that RBC aggregation plays an important role in the dynamic of platelets and leukocytes in vivo.
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Adesão Celular , Agregação Eritrocítica , Microesferas , Microtecnologia/instrumentação , Adesão Celular/efeitos dos fármacos , Dextranos/farmacologia , Agregação Eritrocítica/efeitos dos fármacos , HumanosRESUMO
Red blood cells (RBCs) are the most abundant cells in human blood. Remarkably RBCs 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 main objective of this work is to quantify and characterize RBC aggregates in order to enhance the current understanding of the non-Newtonian behaviour of blood in microcirculation. Suspensions of human blood were flowed and observed in vitro in poly-di-methyl-siloxane (PDMS) microchannels to characterize RBC aggregates. These microchannels were fabricated using standard photolithography methods. Experiments were performed using a micro particle image velocimetry (µPIV) system for shear rate measurements, coupled with a high-speed camera for flow visualization. RBC aggregate sizes were quantified in controlled and measurable shear rate environments for 5, 10 and 15% hematocrit. Aggregate sizes were determined using image processing techniques, while apparent viscosity was measured using optical viscometry. For the samples suspended at 5% H, aggregate size was not strongly correlated with shear rate. For the 10% H suspensions, in contrast, lowering the shear rate below 10 s-1 resulted in a significant increase of RBC aggregate sizes. The viscosity was found to increase with decreasing shear rate and increasing hematocrit, exemplifying the established non-Newtonian shear-thinning behaviour of blood. Increase in aggregation size did not translate into a linear increase of the blood viscosity. Temperature was shown to affect blood viscosity as expected, however, no correlation for aggregate size with temperature was observed. Non-Newtonian parameters associated with power law and Carreau models were determined by fitting the experimental data and can be used towards the simple modeling of blood's non-Newtonian behaviour in microcirculation. This work establishes a relationship between RBC aggregate sizes and corresponding shear rates and one between RBC aggregate sizes and apparent blood viscosity at body and room temperatures, in a microfluidic environment for low hematocrit. Effects of hematocrit, shear rate, viscosity and temperature on RBC aggregate sizes have been quantified.
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Viscosidade Sanguínea , Agregação Eritrocítica , Eritrócitos/fisiologia , Hematócrito , Eritrócitos/citologia , Humanos , Microcirculação , Microfluídica/instrumentação , Microfluídica/métodosRESUMO
Blood, as a non-Newtonian biofluid, represents the focus of numerous studies in the hemorheology field. Blood constituents include red blood cells, white blood cells and platelets that are suspended in blood plasma. Due to the abundance of the RBCs (40% to 45% of the blood volume), their behavior dictates the rheological behavior of blood especially in the microcirculation. At very low shear rates, RBCs are seen to assemble and form entities called aggregates, which causes the non-Newtonian behavior of blood. It is important to understand the conditions of the aggregates formation to comprehend the blood rheology in microcirculation. The protocol described here details the experimental procedure to determine quantitatively the RBC aggregates in microcirculation under constant shear rate, based on image processing. For this purpose, RBC-suspensions are tested and analyzed in 120 x 60 µm poly-dimethyl-siloxane (PDMS) microchannels. The RBC-suspensions are entrained using a second fluid in order to obtain a linear velocity profile within the blood layer and thus achieve a wide range of constant shear rates. The shear rate is determined using a micro Particle Image Velocimetry (µPIV) system, while RBC aggregates are visualized using a high speed camera. The videos captured of the RBC aggregates are analyzed using image processing techniques in order to determine the aggregate sizes based on the images intensities.
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Agregação Eritrocítica/fisiologia , Microfluídica/métodos , Humanos , Microfluídica/instrumentaçãoRESUMO
High speed photography in micro-particle image velocimetry (µPIV) using red blood cells as tracer particles and the use of fluorescing tracer particles (in conjunction with pulsed images) are directly compared by using both methods simultaneously. Measurements are taken on the same blood sample in the same microchip using both methods. This work directly and statistically compares the two methods of µPIV measurement in a controlled in vitro environment for the first time in literature. The pulsed method using fluorescing tracer particles is found to decrease the depth of correlation as expected, and to better represent the shape of the velocity profile. Two methods of velocity characterization are used (single and double parameter) and the pulsed images provide better shape representation in both cases.
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Eritrócitos/citologia , Corantes Fluorescentes , Microesferas , Imagem Molecular/métodos , Reologia/métodos , Animais , SuínosRESUMO
It is desired to understand the effect of alginic acid sodium salt from brown algae (alginate) as a viscosity modifier on the behavior of blood in vitro using a micro-particle image velocimetry (µPIV) system. The effect of alginate on the shape of the velocity profile, the flow rate and the maximum velocity achieved in rectangular microchannels channels are measured. The channels were constructed of polydimethylsiloxane (PDMS), a biocompatible silicone. Porcine blood cells suspended in saline was used as the working fluid at twenty percent hematocrit (Hâ=â20). While alginate was only found to have minimal effect on the maximum velocity and the flow rate achieved, it was found to significantly affect the shear rate at the wall by between eight to a hundred percent.
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Alginatos/farmacologia , Microfluídica/métodos , Microesferas , Reologia/métodos , Animais , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Dimetilpolisiloxanos/química , Ácido Glucurônico/farmacologia , Hematócrito , Ácidos Hexurônicos/farmacologia , Resistência ao Cisalhamento/efeitos dos fármacos , Sus scrofaRESUMO
Micro-particle image velocimetry (µPIV) is used to visualize paired images of micro particles seeded in blood flows. The images are cross-correlated to give an accurate velocity profile. A protocol is presented for µPIV measurements of blood flows in microchannels. At the scale of the microcirculation, blood cannot be considered a homogeneous fluid, as it is a suspension of flexible particles suspended in plasma, a Newtonian fluid. Shear rate, maximum velocity, velocity profile shape, and flow rate can be derived from these measurements. Several key parameters such as focal depth, particle concentration, and system compliance, are presented in order to ensure accurate, useful data along with examples and representative results for various hematocrits and flow conditions.
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Dispositivos Lab-On-A-Chip , Fluxo Sanguíneo Regional/fisiologia , Reologia/métodos , Animais , Tamanho da Partícula , Reologia/instrumentação , SuínosRESUMO
Micro particle image velocimetry (µPIV) is a common method to assess flow behavior in blood microvessels in vitro as well as in vivo. The use of red blood cells (RBCs) as tracer particles, as generally considered in vivo, creates a large depth of correlation (DOC), even as large as the vessel itself, which decreases the accuracy of the method. The limitations of µPIV for blood flow measurements based on RBC tracking still have to be evaluated. In this study, in vitro and in silico models were used to understand the effect of the DOC on blood flow measurements using µPIV RBC tracer particles. We therefore employed a µPIV technique to assess blood flow in a 15 µm radius glass tube with a high-speed CMOS camera. The tube was perfused with a sample of 40% hematocrit blood. The flow measured by a cross-correlating speckle tracking technique was compared to the flow rate of the pump. In addition, a three-dimensional mechanical RBC-flow model was used to simulate optical moving speckle at 20% and 40% hematocrits, in 15 and 20 µm radius circular tubes, at different focus planes, flow rates and for various velocity profile shapes. The velocity profiles extracted from the simulated pictures were compared with good agreement with the corresponding velocity profiles implemented in the mechanical model. The flow rates from both the in vitro flow phantom and the mathematical model were accurately measured with less than 10% errors. Simulation results demonstrated that the hematocrit (paired t tests, p = 0.5) and the tube radius (p = 0.1) do not influence the precision of the measured flow rate, whereas the shape of the velocity profile (p < 0.001) and the location of the focus plane (p < 0.001) do, as indicated by measured errors ranging from 3% to 97%. In conclusion, the use of RBCs as tracer particles makes a large DOC and affects the image processing required to estimate the flow velocities. We found that the current µPIV method is acceptable to estimate the flow rate on the condition that the measurement takes place at the equatorial plane of the vessel. Otherwise, it is not an appropriate method to estimate the shape of the velocity profile.
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Hemodinâmica , Modelos Biológicos , Fenômenos Ópticos , Reologia/métodos , Animais , Fenômenos Biomecânicos , Velocidade do Fluxo Sanguíneo , Biologia Computacional , Eritrócitos/citologia , SuínosRESUMO
To elucidate the relationship between microscopic red blood cell (RBC) interactions and macroscopic rheological behavior, we propose a two-dimensional particle model capable of mimicking the main characteristics of RBC aggregation kinetics. The mechanical model of RBCs sheared in Couette flow is based on Newton law. We assumed a hydrodynamic force to move particles, a force to describe aggregation and an elasticity force. The role of molecular mass and concentration of neutral polymers on aggregation [Neu, B., and H. J. Meiselman. Biophys. J. 83:2482-2490, 2002] could be mimicked. Specifically, it was shown that for any shear rate (SR), the mean aggregate size (MAS) grew with time until it reached a constant value, which is consistent with in vitro experiments. It was also demonstrated that we could mimic the modal relationship between MAS and SR and the occurrence of maximum aggregation at about 0.1 s(-1). As anticipated, simulations indicated that an increase in aggregation force augmented MAS. Further, augmentation of the depletion layer thickness influenced MAS only for SR close to zero, which is a new finding. To conclude, our contribution reveals that the aggregation force intensity and SR influence the steady state MAS, and that the depletion and layer thickness affect the aggregation speed.
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Agregação Eritrocítica/fisiologia , Eritrócitos/citologia , Eritrócitos/fisiologia , Modelos Cardiovasculares , Tamanho Celular , Células Cultivadas , Simulação por Computador , Módulo de Elasticidade/fisiologia , Humanos , Cinética , Tamanho da Partícula , Resistência ao Cisalhamento/fisiologia , Estresse MecânicoRESUMO
Power Doppler imaging (PDI) has been shown to be influenced by the wall filter when assessing arterial stenoses. Real-time 3-D Doppler imaging may likely become a widespread practice in the near future, but how the wall filter could affect PDI during the cardiac cycle has not been investigated. The objective of the study was to demonstrate that the wall filter may produce unexpected major signal losses in real-time 3-D PDI. To test our hypothesis, we first validated binary images obtained from analytical simulations with in vitro PDI acquisitions performed in a tube under pulsatile flow conditions. We then simulated PDI images in the presence of a severe stenosis, considering physiological conditions by finite element modeling. Power Doppler imaging simulations revealed important signal losses within the lumen area at different instants of the flow cycle, and there was a very good concordance between measured and predicted PDI binary images in the tube. Our results show that the wall filter may induce severe PDI signal losses that could negatively influence the assessment of vascular stenosis. Clinicians should therefore be aware of this cause of signal loss to properly interpret power Doppler angiographic images.
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Arteriopatias Oclusivas/diagnóstico por imagem , Artérias/diagnóstico por imagem , Imageamento Tridimensional/normas , Processamento de Sinais Assistido por Computador , Ultrassonografia Doppler em Cores/normas , Simulação por Computador , Análise de Elementos Finitos , Humanos , Imageamento Tridimensional/métodos , Técnicas In Vitro , Modelos Biológicos , Ultrassonografia Doppler em Cores/métodosRESUMO
We propose a method for calculating directly total body water (TBW) volumes (V (t)) from whole body resistance extrapolated at infinite frequency (R (infinity)) using a XITRON 4200 impedance meter. Mean TBW resistivities for men and women were determined from measurements of R (infinity) and fat-free mass (FFM(d)) measured by DXA in 58 healthy subjects assuming an average hydration coefficient of 73.2%. Mean differences between V (t) measured by our new method and those deduced from DXA data were +0.11 +/- 1.61 L for women and +0.13 +/- 2.16 L for men. For validation, this method was tested with the same resistivities against a 2nd group of 16 volunteers and the mean difference between V (t) from impedance and DXA was -0.80 +/- 1.43 L. Since the resistance at 50 kHz (R (50)) was found to be equal, in average, to 1.230 R (infinity) for men and 1.223 R (infinity) for women, this method can also be applied at 50 kHz with a similar accuracy by estimating R (infinity) from R (50). When our new method was applied to the monitoring of water loss during 28 dialysis runs performed on 13 patients, it predicted a mean water loss equal to 94% of ultrafiltered volume.
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Água Corporal , Impedância Elétrica , Análise Espectral/métodos , Absorciometria de Fóton/métodos , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Matemática , Pessoa de Meia-Idade , Diálise Renal/métodosRESUMO
Extracellular (ECW) and intracellular (ICW) volumes were measured using both segmental and wrist-ankle (W-A) bioimpedance spectroscopy (5-1000 kHz) in 15 healthy subjects (7 men, 8 women). In the 1st protocol, the subject, after sitting for 30 min, laid supine for at least 30 min. In the second protocol, the subject, who had been supine for 1 hr, sat up in bed for 10 min and returned to supine position for another hour. Segmental ECW and ICW resistances of legs, arms and trunk were measured by placing four voltage electrodes on wrist, shoulder, top of thigh and ankle and using Hanai's conductivity theory. W-A resistances were found to be very close to the sum of segmental resistances. When switching from sitting to supine (protocol 1), the mean ECW leg resistance increased by 18.2%, that of arm and W-A by 12.4%. Trunk resistance also increased but not significantly by 4.8%. Corresponding increases in ICW resistance were smaller for legs (3.7%) and arm (-0.7%) but larger for the trunk (21.4%). Total body ECW volumes from segmental measurements were in good agreement with W-A and Watson anthropomorphic correlation. The decrease in total ECW volume (when supine) calculated from segmental resistances was at 0.79 l less than the W-A one (1.12 l). Total ICW volume reductions were 3.4% (segmental) and 3.8% (W-A). Tests of protocol 2 confirmed that resistance and fluid volume values were not affected by a temporary position change.
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Água Corporal/fisiologia , Líquido Extracelular/fisiologia , Deslocamentos de Líquidos Corporais/fisiologia , Líquido Intracelular/fisiologia , Pletismografia de Impedância/métodos , Postura/fisiologia , Análise Espectral/métodos , Adulto , Tornozelo , Antropometria/métodos , Braço/fisiologia , Compartimentos de Líquidos Corporais/fisiologia , Impedância Elétrica , Espaço Extracelular/fisiologia , Feminino , Humanos , Espaço Intracelular/fisiologia , Perna (Membro)/fisiologia , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tórax/fisiologia , PunhoRESUMO
Bioimpedance spectroscopy (BIS) permits evaluation of extra- and intracellular fluid volumes in patients. We wished to examine whether this technique, used in combination with hematocrit measurement, can reliably monitor fluid transfers during dialysis. Ankle to wrist BIS measurements were collected during 21 dialysis runs while hematocrit was continuously monitored in the blood line by an optical device. Extracellular (ECW) and intracellular (ICW) water volumes were calculated using Hanai's electrical model of suspensions. Plasma volume variations were calculated from hematocrit, and changes in interstitial volume were calculated as the difference between ECW and plasma volume changes. Because accuracy of ICW was too low, changes in ICW were calculated as the difference between ultrafiltered volume and ECW changes. Total body water (TBW) volumes calculated pre- and postdialysis were, respectively, 3.25+/-3.2 and 1.95+/-2.5 liters lower on average than TBW given by Watson et al.'s correlation. Average decreases in fluid compartments expressed as percentage of ultrafiltered volume were as follows: plasma, 18%; interstitial, 28%, and ICW, 54%. When the ultrafiltered volume was increased in a patient in successive runs, the relative contributions of ICW and interstitial fluid were augmented so as to reduce the relative drop in plasma volume.