Lithium Distribution in Red Blood Cells and Plasma: NMR Studies of Rat Blood

To understand the interaction of lithium (Li⁺) with a coadministered drug in both the blood and the brain, we have treated rats with either Li⁺ alone or Li⁺ and a codrug. In this paper we address the important problem of quantitation of intra and extracellular Li⁺ ion contents in blood by the ⁷Li-NMR technique and the use of a shift reagent (SR). Although Li⁺ can be studied by atomic absorption techniques, these techniques involve tedious separation of intra- and extracellular components prior to chemical analysis. Magnetic resonance studies on rat blood, in the dose range of 0.5 to 10 meq/kg, indicate that the intracellular red blood cell Li⁺ predominates in the lower dose range of 0.5–1.0 meq/kg. As the lithium dose increases, a significantly larger amount of Li⁺ accumulates in the extracellular volume. Our studies on a number of animals at various doses of LiCl indicate that ⁷Li-NMR of blood samples provide a reliable, noninvasive quantification of red blood cell and plasma Li⁺ concentrations. The NMR method was further used to study the effect of coadministered drugs such as thioridazine on the intra- and extracellular Li⁺ concentration of RBCs.     

Numerical simulation of the red blood cell aggregation and deformation behaviors in ultrasonic field

The objective of this paper is to propose an immersed boundary lattice Boltzmann method (IB-LBM) considering the ultrasonic effect to simulate red blood cell (RBC) aggregation and deformation in ultrasonic field. Numerical examples involving the typical streamline, normalized out-of-plane vorticity contours and vector fields in pure plasma under three different ultrasound intensities are presented. Meanwhile, the corresponding transient aggregation behavior of RBCs, with special emphasis on the detailed process of RBC deformation, is shown. The numerical results reveal that the ultrasound wave acted on the pure plasma can lead to recirculation flow, which contributes to the RBCs aggregation and deformation in microvessel. Furthermore, increasing the intensity of the ultrasound wave can significantly enhance the aggregation and deformation of the RBCs. And the formation of the RBCs aggregation leads to the fluctuated and dropped vorticity value of plasma in return.     

A non-invasive method for the assessment of hemostasis in vivo by using dynamic light scattering

We present a new non-invasive method for assessing hemostasis in vivo. This method is based on the analysis of the movement characteristics of red blood cells (RBCs) during blood stasis condition. Stasis is intermittently induced by occlusion of arterial blood flow at the finger root. We assumed that under zero flow conditions, RBC movement is driven mostly by Brownian motion, and we characterized the RBC movement by utilizing the dynamic light scattering (DLS) technique in vivo. We found that during the stasis the RBCs diffusion coefficient in plasma decreases. We speculate that the RBC diffusion coefficient is most strongly related to endothelial and hemostatic activity. This assumption is supported by our findings that RBC movement, being expressed through the characteristics of the measured DLS signal, is correlative to the biological age and also is related to the coagulation factors. This new method can serve as a new diagnostic and research tool for the assessment of hemostasis and vascular function.     

Estimation of red blood cell aggregate velocity during sedimentation using the Hough transform

A method for velocity estimation of sedimenting three-dimensional (3D) red blood cell (RBC) aggregates by means of an image processing technique is proposed. Successive images of RBC suspension near the wall of a container reveal rouleaux formation, sedimentation of 3D RBC aggregates and formation of the deposit of the cells. Plots of the position versus time for the 3D RBC aggregates were extracted by a processing of successive images of the suspension. The plots exhibit a quasi-linear structures in noisy background. With the use of the Hough transform the detection of the slope of the structures was performed and the velocity of the aggregates was estimated. To show the potential of the method spatio-temporal dependence of the aggregate velocity is presented for RBCs in plasma, RBCs in Dextran and for hardened cells at haematocrit 5%.     

舌诊归一化反射光谱用于红细胞数无创测量

利用舌诊归一化反射光谱对人体血液中红细胞总数(RBC)进行了无创检测研究.采集240名志愿者舌尖反射光谱并进行反射率归一化同时用数码相机记录志愿者舌象.将样本分为两组:校正集和预测集,以血液成分含量生化分析值为参考,建立红细胞总数与光谱数据的偏最小二乘回归(PLSR)预测模型.所建模型的相关系数为0.991,用所建模型...     

Numerical Study on the Dynamics and Oxygen Uptake of Healthy and Malaria-Infected Red Blood Cells

Red blood cells (RBCs) are very important due to their role of oxygen transport from lungs. As the malaria parasite grows in the malaria-infected red blood cells (IRBCs), the properties of the cells change. In the present work, the oxygen uptake by RBCs and IRBCs at the pulmonary capillaries is simulated using a numerical technique based on the two-dimensional immersed interface method. The results for the oxygen uptake by a stationary single RBC have fair agreements with the previously reported results. The numerical results show that the malaria infection could significantly cause deterioration on the oxygen uptake by red blood cells. The results also suggest that the oxygen uptake by individual stationary RBC/IRBC would not be significantly affected by the neighboring cells provided the separation distance is about the dimension of the cell. Furthermore, it appears that the oxygen uptake by both RBCs and IRBCs is dominated by mass diffusion over the convection although the Peclet number is of the order of unity.     

Mechanical behavior of pathological and normal red blood cells in microvascular flow based on modified level-set method

The research of the motion and deformation of the RBCs is important to reveal the mechanism of blood diseases. A numerical method has been developed with level set formulation for elastic membrane immersed in incompressible fluid. The numerical model satisfies mass and energy conservation without the leaking problems in classical Immersed Boundary Method(IBM), at the same time, computing grid we used can be much smaller than the general literatures. The motion and deformation of a red blood cell(including pathological normal status) in microvascular flow are simulated. It is found that the Reynolds number and membrane's stiffness play an important role in the transmutation and oscillation of the elastic membrane. The normal biconcave shape of the RBC is propitious to create high deformation than other pathological shapes. With reduced viscosity of the interior fluid both the velocity of the blood and the deformability of the cell reduced. With increased viscosity of the plasma both the velocity of the blood and the deformability of the cell reduced. The tank treading of the RBC membrane is observed at low enough viscosity contrast in shear flow. The tank tread fixed inclination angle of the cell depends on the shear ratio and viscosity contrast, which can be compared with the experimental observation well.     

NMR Studies of Intra- and Extracellular Red Blood Cell Lithium by Transverse Relaxation Measurements and Shift Reagents

Lithium salts are used in the treatment and prophylaxis of bipolar or mood disorders. The mechanism of action by which the cation exerts its therapeutic influence is unknown. A knowledge of brain Li concentration, its distribution in the brain, and its properties in the cellular microenvironment may have a strong influence on the understanding of Li function. The differentiation of lithium in the intra and extracellular environments has been achieved in a noninvasive manner in red blood cell (RBC) model. The two distinct transverse relaxation (T₂) components have been observed in the blood sample drawn from lithium treated rats. These results indicate two different environments for Li with a fast (T_2f) and a slow (T_2s) component in the RBC model corresponding fractions that contribute to each relaxation component. The results compare well with the intra- and extracellular RBC lithium measured using shift reagents. Our studies indicate that the T₂ method has utility in estimating the intracellular Li in systems that exhibit similar T₂ behavior. The studies performed at different Li doses in the rat model indicate that the method may have utility in following a wide range of intracellular Li.     

Experimental ultrasound characterization of red blood cell aggregation using the structure factor size estimator

The frequency dependence of the ultrasonic backscattering coefficient (BSC) was studied to assess the level of red blood cell (RBC) aggregation. Three monoelement focused wideband transducers were used to insonify porcine blood sheared in a Couette flow from 9 to 30 MHz. A high shear rate was first applied to promote disaggregation. Different residual shear rates were then used to promote formation of RBC aggregates. The structure factor size estimator (SFSE), a second-order data reduction model based on the structure factor, was applied to the frequency-dependent BSC. Two parameters were extracted from the model to describe the level of aggregation at 6% and 40% hematocrits: W, the packing factor, and D the aggregate diameter, expressed in number of RBCs. Both parameters closely matched theoretical values for nonaggregated RBCs. W and D increased during aggregation with stabilized values modulated by the applied residual shear rate. Furthermore, parameter D during the kinetics of aggregation at 6% hematocrit under static conditions correlated with an optical RBC aggregate size estimation from microscopic images (r(2)=0.76). To conclude, the SFSE presents an interesting framework for tissue characterization of partially correlated dense tissues such as aggregated RBCs.     

Assessment of accuracy of the structure-factor-size-estimator method in determining red blood cell aggregate size from ultrasound spectral backscatter coefficient

A computer simulation study to produce ultrasonic backscatter coefficients (BSCs) from red blood cell (RBC) clusters is discussed. The simulation algorithm is suitable for generating non-overlapping, isotropic, and fairly identical RBC clusters. RBCs were stacked following the hexagonal close packing (HCP) structure to form a compact spherical aggregate. Such an aggregate was repeated and placed randomly under non-overlapping condition in the three-dimensional space to mimic an aggregated blood sample. BSCs were computed between 750 KHz and 200 MHz for samples of various cluster sizes at different hematocrits. Magnitudes of BSCs increased with mean aggregate sizes at low frequencies (<20 MHz). The accuracy of the structure-factor-size-estimator (SFSE) method in determining mean aggregate size and packing factor was also examined. A good correlation (R(2) ≥ 0.94) between the mean size of aggregates predicted by the SFSE and true size was found for each hematocrit. This study shows that for spherical aggregates there exists a region for each hematocrit where SFSE works most accurately. Typically, error of SFSE in estimating mean cluster size was <20% for dimensions between 14 and 17 μm at 40% hematocrit. This study suggests that the theoretical framework of SFSE is valid under the assumption of isotropic aggregates.     

Lithium spectroscopic imaging of rat brain at therapeutic doses

Since the discovery that lithium (Li) is efficacious for the treatment of manic depressive illness, the brain Li distribution of mammals treated with lithium has been of interest. However, the spatial relationship of lithium in the brain regions to its function remains largely unknown. Knowledge of Li distribution in the brain is necessary to localize its action in the brain. Both the therapeutic and neurotoxic side effects of Li are centered mainly in the central nervous system and hence there is considerable interest in understanding the extent of lithium penetration into the central nervous system. The mechanism by which neurotoxic side effects are generated is not known and may, in part, be related to the particular distribution of lithium in the brain. The regional specificity in lithium's brain distribution could underlie important steps on its action. Li levels in various brain regions for autopsied rats and humans have been reported. However, many results are conflicting due to ion redistribution at death or during sample preparation. A direct nondestructive measurement of Li levels in the brain where the drug exerts its effects is certainly desirable. Because magnetic resonance technique can be used to observe Li, it can be an appropriate method to monitor and map the distribution in the brain. The application of MR technology to rat brain regions has provided information on lithium distribution in a non-invasive manner. The earlier development work at lower field strengths provided brain lithium information at high dose of Li administration. Here we demonstrate the feasibility of quantitative spectroscopic imaging on rat brain under therapeutic doses.     

High‐resolution Raman imaging reveals spatial location of heme oxidation sites in single red blood cells of dried smears

Raman confocal microscopy with 488 nm excitation wavelength supported with atomic force microscopy (AFM), scanning near‐field optical microscopy (SNOM) and UV–Vis spectrometry was used to investigate air‐dried erythrocytes (red blood cells, RBCs) in whole human blood smears. The central internal part of the cell was dominated by the laser‐induced O₂ dissociated oxyHb form as evidenced by the Fe²⁺ marker band appearing at 1356 cm⁻¹. The existence of a thin outer layer of hemoglobin in the periphery of RBCs was assigned to hemichrome. Evidence for hemichrome includes the oxidation state marker band appearing at 1376 cm⁻¹, the absence of FeO₂ band at 570 cm⁻¹ and a UV–Vis spectrum consistent with hemichrome. This is the first time that distributions of Fe²⁺/Fe³⁺ hemes inside the single RBC have been reported. The outer layer formation of hemichrome was additionally studied when RBCs were in contact with leucocytes and carotenoids of blood plasma. Copyright © 2014 John Wiley & Sons, Ltd.     

Application of phosphoenolpyruvate into canine red blood cell cryopreservation with hydroxyethyl starch

Phosphoenolpyruvate (PEP) is a phosphorylated glycolytic intermediate that can penetrate the RBC membrane and be metabolized to 2,3-DPG and ATP. In this study, we evaluated the effects of PEP treatment on canine red blood cells (RBCs) cryopreserved with 12.5% (w/v) HES. RBCs were incubated for 30, 60, and 90 min at 37 degrees C with PEP solution containing 60 mM mannitol, 30 mM sodium chloride, 25 mM glucose, 1 mM adenine and 50 mM PEP (340 m osm/kg), pH 6.0 and then cryopreserved in liquid nitrogen with 12.5% (w/v) HES for 2 weeks. 2,3-DPG and saline stabilities of the PEP treated groups were increased and osmotic fragility indices were significantly decreased compared to the untreated control group. There were no differences in 2,3-DPG levels within the PEP treated groups with different PEP incubation times. These results suggest that PEP treatment may be beneficial for the cryopreservation of canine RBCs with HES.     

Modeling the frequency dependence (5-120 MHz) of ultrasound backscattering by red cell aggregates in shear flow at a normal hematocrit

The frequency dependence of the ultrasound signal backscattered by blood in shear flow was studied using a simulation model. The ultrasound backscattered signal was computed with a linear model that considers the characteristics of the ultrasound system and tissue acoustic properties. The tissue scattering properties were related to the position and shape of the red blood cells (RBCs). A 2D microrheological model simulated the RBC dynamics in a Couette shear flow system. This iterative model, described earlier [Biophys. J. 82, 1696-1710 (2002)], integrates the hydrodynamic effect of the flow, as well as adhesive and repulsive forces between RBCs. RBC aggregation was simulated at 40% hematocrit and shear rates of 0.05-2 s(-1). The RBC aggregate sizes ranged, on average, from 3.3 RBCs at 2 s(-1) to 33.5 cells at 0.05 s(-1). The ultrasound backscattered power was studied at frequencies between 5-120 MHz and insonification angles between 0-180 degrees. At frequencies below approximately 30 MHz, the ultrasound backscattered power increased as the shear rate was decreased and the size of the aggregates was raised. A totally different scattering behavior was noted above 30 MHz. Typical spectral slopes of the backscattered power (log-log scale) between 5-25 MHz equaled 3.8, whereas slopes down to 0.6 were measured at 0.05 s(-1), between 40-60 MHz. The ultrasound backscattered power was shown to be angle dependent at low frequencies (5-25 MHz). The anisotropy persisted at high frequencies (>25 MHz) for small aggregates (at 2 s(-1)). In conclusion, this study sheds some light on the blood backscattering behavior with an emphasis on the non-Rayleigh regime. Additional experimental studies may be necessary to validate the simulation results, and to fully understand the relation between the ultrasound backscattered power, level of RBC aggregation, shear rate, frequency, and insonification angle.     

Time-dependent surface adhesive force and morphology of RBC measured by AFM

Atomic force microscopy (AFM) is a rapidly developing tool recently introduced into the evaluation of the age of bloodstains, potentially providing legal medical experts useful information for forensic investigation. In this study, the time-dependent, morphological changes of red blood cells (RBC) under three different conditions (including controlled, room-temperature condition, uncontrolled, outdoor-environmental condition, and controlled, low-temperature condition) were observed by AFM, as well as the cellular viscoelasticity via force-vs-distance curve measurements. Firstly, the data indicate that substrate types have different effects on cellular morphology of RBC. RBC presented the typical biconcave shape on mica, whereas either the biconcave shape or flattened shape was evident on glass. The mean volume of RBCs on mica was significantly larger than that of cells on glass. Surprisingly, the adhesive property of RBC membrane surfaces was substrate type-independent (the adhesive forces were statistically similar on glass and mica). With time lapse, the changes in cell volume and adhesive force of RBC under the controlled room-temperature condition were similar to those under the uncontrolled outdoor-environmental condition. Under the controlled low-temperature condition, however, the changes in cell volume occurred mainly due to the collapse of RBCs, and the curves of adhesive force showed the dramatic alternations in viscoelasticity of RBC. Taken together, the AFM detections on the time-dependent, substrate type-dependent, environment (temperature/humidity)-dependent changes in morphology and surface viscoelasticity of RBC imply a potential application of AFM in forensic medicine or investigations, e.g., estimating age of bloodstain or death time.     

Some comments on the effectiveness of the therapy for Β-thalassemia

In the present study, twenty-five samples of red blood cells (RBCs) and two samples of lyophilized serum, drawn from thirteen patients with -thanassemia major, have been examined by Mössbauer spectroscopy. All these patients undergo long-term therapy by regular blood transfusion and deferoxamine. The samples were obtained at the end of one cycle of treatments, before the successive blood transfusion. The results show, within the experimental errors, that the ferritin-like iron appears to be absent in the RBCs of the patients but it is still present in the serum.On leave from Tsinghua University, Beijing, PR China.On leave fro East China Normal University, Shanghai, PR China.     

Accurate coarse-grained modeling of red blood cells

We develop a systematic coarse-graining procedure for modeling red blood cells (RBCs) using arguments based on mean-field theory. The three-dimensional RBC membrane model takes into account the bending energy, in-plane shear energy, and constraints of fixed surface area and fixed enclosed volume. The coarse-graining procedure is general, it can be used for arbitrary level of coarse-graining and does not employ any fitting parameters. The sensitivity of the coarse-grained model is investigated and its behavior is validated against available experimental data and in dissipative particle dynamics (DPD) simulations of RBCs in capillary and shear flows.     

Assessment of scanner performance and normalization of estimated relaxation rate values

The purpose of this study was to develop and test a method for the assessment of Magnetic Resonance (MR) scanner performance suitable for routine brain MR studies and for normalization of calculated relaxation times. We hypothesized that regular monitoring of machine performance changes could provide a helpful normalization tool for calculating tissue MR parameters, thus contributing to support their use for longitudinal and comparative studies of both normal and diseased tissues.The method is based on the acquisition of phantom images during routine brain studies with standard spin-echo sequences. MR phantom and brain tissue parameters were used to assess the influence of machine related changes on relaxation parameter estimates. Experimental results showed that scanner performance may affect relaxation rate estimates. Phantom and in vivo results indicate that the correction method yields a reduction in variability of estimated phantom R1 values up to 29% and of R1 for different brain structures up to 17%. These findings support the validity of using brain coil phantoms for routine system monitoring and correction of tissue relaxation rates.     

Effects of red blood cell aggregates dissociation on the estimation of ultrasound speckle image velocimetry

Ultrasound speckle image of blood is mainly attributed by red blood cells (RBCs) which tend to form RBC aggregates. RBC aggregates are separated into individual cells when the shear force is over a certain value. The dissociation of RBC aggregates has an influence on the performance of ultrasound speckle image velocimetry (SIV) technique in which a cross-correlation algorithm is applied to the speckle images to get the velocity field information. The present study aims to investigate the effect of the dissociation of RBC aggregates on the estimation quality of SIV technique. Ultrasound B-mode images were captured from the porcine blood circulating in a mock-up flow loop with varying flow rate. To verify the measurement performance of SIV technique, the centerline velocity measured by the SIV technique was compared with that measured by Doppler spectrograms. The dissociation of RBC aggregates was estimated by using decorrelation of speckle patterns in which the subsequent window was shifted as much as the speckle displacement to compensate decorrelation caused by in-plane loss of speckle patterns. The decorrelation of speckles is considerably increased according to shear rate. Its variations are different along the radial direction. Because the dissociation of RBC aggregates changes ultrasound speckles, the estimation quality of SIV technique is significantly correlated with the decorrelation of speckles. This degradation of measurement quality may be improved by increasing the data acquisition rate. This study would be useful for simultaneous measurement of hemodynamic and hemorheological information of blood flows using only speckle images.     

Mössbauer spectroscopy in biomedical physics research

Several applications of Mössbauer spectroscopy (MS) as an analytical tool in research on biomedical physics are reviewed: (1) The evaluation of treatments for some diseases such as thalassemia, iron-overload disease, high altitude polycythemia. (2) Medical research on the effects of environmental factors on the human body, for example, the effects of electromagnetic radiation on human red blood cells (RBCs). Some advantages and weaknesses of MS, a new application of the Mössbauer effect, cancer therapy, and some possible applications such as monitoring the RBCs of the patients before, during, and after surgical operation, are discussed.