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Effects of the hydrodynamic environment and shear protectants on survival of erythrocytes in suspension.

Survival of media-suspended porcine erythrocytes exposed to various hydrodynamic environments was investigated with and without such shear protectant additives as bovine serum albumin, dextran and the non-ionic surfactant Pluronic F68. Erythrocytes provided a model cell population with cells of a uniform size, metabolic state and shear tolerance. Because the cells were non-growing, any shear adaptation effects were avoided. Cell lysis was followed by microscopic counts or release of haemoglobin. The cells were susceptible to agitation damage in unaerated shake flasks agitated at 100 rpm or greater. Relative to additives-free operation, the presence of 0.1% (w/v) dextran or albumin prolonged cell survival, but Pluronic F68 actually enhanced cell lysis in flasks agitated at 100 rpm. The protective effect of the additives depended on the hydrodynamic conditions. The protective effect of albumin was demonstrated also in aerated conditions in a split-cylinder airlift bioreactor (aspect ratio of 8.8; riser-to-downcomer cross-sectional area ratio of 1.0; specific power input of 0.34 W m-3). Comparison of the cell lysis characteristics in the airlift device and the best case performance of the shake flask showed longer survival in the flask (100 rpm); however, the length of survival in the reactor (approx. 70 h) was sufficient for practical purposes. In all cases, the cell lysis pattern conformed initially to zero-order dependence in cell concentration, becoming first-order after varying degrees of exposure to hydrodynamic forces. Fatigue failure of cells was inferred.

Animals↗

On the hydrodynamic analysis of macromolecular conformation.

Hydrodynamics provides a powerful complementary role to the traditional "high resolution" techniques for the investigation of macromolecular conformation, especially in dilute solution, conditions which are generally inaccessible to other structural techniques. This paper describes the state of art of hydrodynamic representations for macromolecular conformation, in terms of (1) simple but straightforward ellipsoid of revolution modelling; (2) general triaxial ellipsoid modelling; (3) hydrodynamic bead modelling; (4) the ability, especially for polydisperse macromolecular systems, to distinguish between various conformation types; (5) analysis of macromolecular flexibility.

Journal Article↗

Superposition of hydrodynamic forces on a hair bundle.

Vertebrates sense sound, orientation, and motion by means of bundles of microscopic sensory hairs that protrude from the surfaces of receptor (hair) cells. To determine the effects of the sensory epithelium, tectorial structures, and fluids on the motions of hair bundles, we examine a class of mathematical models in which hair-cell organs are represented as a system of rigid mechanical structures surrounded by fluid. The epithelium and tectorial structures are represented by rigid basal and tectorial plates, respectively; the hair bundle by a rigid body hinged to the basal plate. When the displacements of these structures are small, the equations of motion for the fluid are predominately linear. Therefore, both the fluid velocity and the force of fluid origin on the body can be expressed as a sum of components; each component results from motion of a single structure while all others are stationary. This analysis leads to a network description of the motion of the rigid body in which hydrodynamic forces are segregated from mechanical forces. The separation of hydrodynamics and mechanics not only clarifies the effects of fluids on motion but also minimizes the number of hydrodynamic computations needed to analyze models of hair-bundle motion.

Animals↗

Prevention of experimental autoimmune myocarditis by hydrodynamics-based naked plasmid DNA encoding CTLA4-Ig gene delivery.

BACKGROUND: Rat experimental autoimmune myocarditis (EAM) is a T cell-mediated disease that resembled the giant cell myocarditis seen in humans. Soluble CTLA4 improves some autoimmune diseases by blocking costimulatory signals on T cell. We investigated the effect of hydrodynamics-based naked plasmid DNA encoding CTLA4-immunoglobulin (Ig) gene delivery. METHODS AND RESULTS: Lewis rats were immunized with cardiac myosin and treated with hydrodynamic-based transfection, namely a rapid tail vein injection of a large volume of pCAGGS encoding CTLA4-Ig chimera solution on Day 0. The vector-derived CTLA4-Ig mRNA expressions were mainly detected in the liver and plasma CTLA4-Ig protein levels were maintained at about 2 mug/mL during the experiment period. On Day 17, the ratio of heart to body weight, the amount of mRNA of atrial natriuretic peptide, and the inflammatory areas in CTLA4 group were significantly lower than in the control group treated with empty plasmid. Maximum rate of intraventricular pressure rise and decline (dP/dT), minimum dP/dT, left ventricular end-diastolic pressure, and central venous pressure improved significantly after treatment with CTLA4-Ig. On Day 14, expressions of IL-2 in popliteal lymph nodes in the CTLA4-Ig group were significantly lower than in the control group. CONCLUSION: Hydrodynamics-based transfection of plasmid encoding CTLA4-Ig chimera dramatically prevented EAM.

Abatacept↗

Hydrodynamic properties of oxidized extracellular polysaccharides from Erwinia chrysanthemi spp.

The molecular weights of the native polysaccharides of Erwinia chrysanthemi strains range from 1.8 to 7.1 x 10(6) and their hydrodynamic properties are those of polydisperse, polyanionic biopolymers with pseudoplastic, non-thixotropic flow characteristics in aqueous solutions. The effect on the hydrodynamic properties of the polysaccharides by adding carboxyl groups to increase the charge density is studied, with particular reference to their molecular weight (MW), viscosity and conformation. In general, it is found that periodate oxidation of the extracellular polysaccharides of E. chrysanthemi strains, Ech9Sm6 and Ech6S+, introduces little change in the hydrodynamic properties of the resulting polyaldehydes. However, bromine oxidation at neutral pH of the polyaldehydes results in polycarboxylate biopolymers that show significant reduction in MW and viscosity, but they are still characteristic polyanions.

Aldehydes↗

Expansion and hydrodynamic properties of cellulose-stainless steel powder composite matrix for expanded bed adsorption.

For better understanding the influences of solid phase properties on the performance of the expanded bed, the expansion and hydrodynamic properties of cellulose-stainless steel powder composite matrix with a series of densities was investigated and analyzed in an expanded bed. Two kinds of matrix particle diameter fractions, the small one (60-125 microm) and the large (125-300 microm), were used in the present work. In general, the expansion factors decreased obviously with the increase of matrix density. A linear relation between the mean density of matrix and superficial velocity at expansion factor of 2.5 was found for same series of matrices. The Richardson-Zaki equation could correlate the bed expansion and operation fluid velocity for all matrices tested. The theoretical prediction of correlation parameters (the terminal settling velocity U(t) and expansion index n) was improved with the modification of equations in the literature. The residence time distributions were investigated to characterize the hydrodynamic property in expanded bed. Compared with three evaluation factors (the height equivalent of theoretical plate, Bo number and axial distribution coefficient D(ax)), the results indicated that D(ax) is the best parameter to analyze the bed stability of expanded bed under various operation conditions and matrix properties. In addition, it was found that fluid velocity is the most essential factor to influence the hydrodynamic properties in the bed. A linear relation between the D(ax) and superficial fluid velocity for all matrices tested was established.

Adsorption↗

Hydrodynamic fragmentation of nanoparticle aggregates at orthokinetic coagulation.

Hydrodynamic forces on a doublet of large particles or aggregates during sedimentation cause the non-inertial fragmentation of the doublet, if the doublet Reynolds number and Stokes number are small. In lio-dispersed systems, this non-inertial fragmentation is known to promote the wet classification of large particles whereas the small particles are aggregated (in the secondary minimum) and hence cannot be separated despite the electrostatic repulsion. In aero-dispersed systems with negligible electrostatic repulsion, it is possible to have a narrower separation between two interacting particles so that the attractive surface forces such as van der Waals force could increase in orders of magnitude. As a result, the doublet fragmentation by the aerodynamic detaching force becomes very difficult or even impossible in sedimentation at a small Re. However, this study shows that, when extending above analysis for the interacting fractal aggregates of nanoparticles in a suspension, it is still possible to have aggregate (doublet of two nano-aggregates) fragmentation by the aerodynamic detaching force because the surface forces for nanoparticle contact between two aggregates may be in orders of magnitude smaller than that for micron-sized particles. Even with multiple contacts between two interacting nanoparticle aggregates, this prediction of aggregate fragmentation may still be valid because the contacts may break step by step due to the aggregate rolling along each other caused by a short-range aerodynamic interaction during their differential settling. The aerodynamics of aero-dispersed nanoparticle aggregates is analogous to the hydrodynamics of lio-dispersed solid particles. Therefore the hydrodynamic fragmentation model may be used to partially interpret the stability of nanoparticle fluidization process. Our model indicates that, without continued doublet fragmentation of nano-aggregates, the fluidized nanoparticle suspension would be de-fluidized within minutes via the cascading aggregation. However, the experimental evidence of a sustained operation of fluidization of nanoparticle aggregates over a very long time period without changing the size of nanoparticle aggregates indicates that there is a dynamic balance between the aggregation and fragmentation of nano-aggregates in nanoparticle fluidization. The prediction of critical size of nanoparticle aggregates caused by fragmentation in stabilized fluidization agrees with our in situ measurements.

Kinetics↗

Hydrodynamic radius of fractal clusters.

The Kirkwood-Riseman theory has been used to derive an analytical formula for the evaluation of the hydrodynamic radii of fractal clusters. The proposed relation is based on knowledge of the particle-particle correlation function and can be applied to clusters containing any number of particles larger than 4. The calculated values of the hydrodynamic radius are compared with the results of other theoretical approaches proposed in the literature, as well as several experimental data. Finally, the developed relation has been used in connection with a population balance equation model that computes the clusters' mass distribution to estimate the average hydrodynamic radius for a population of fractal clusters of different sizes. The obtained results have been compared to suitable experimental data for a silica colloidal suspension aggregating under reaction-limited conditions.

Journal Article↗

Hydrodynamic characterization of the DEAD-box RNA helicase DbpA.

The Escherichia coli DEAD-box protein A (DbpA) belongs to the highly conserved superfamily-II of nucleic acid helicases that play key roles in RNA metabolism. A central question regarding helicase activity is whether the process of coupling ATP hydrolysis to nucleic acid unwinding requires an oligomeric form of the enzyme. We have investigated the structural and functional properties of DbpA by multi-angle laser light-scattering, size-exclusion chromatography, analytical ultracentrifugation, chemical cross-linking and hydrodynamic modeling. DbpA is monomeric in solution up to a concentration of 25 microM and over the temperature range of 4 degrees C to 22 degrees C. Binding of neither nucleotide (ATP or ADP) nor peptidyl transferase center (PTC) RNA, the presumed physiological RNA substrate, favor oligomerization. The hydrodynamic parameters were used together with hydrodynamic bead modeling and structural homology in conjunction with ab initio structure prediction methods to define plausible shapes of DbpA. Collectively, the results favor models where DbpA functions as an active monomer that possesses two distinct RNA binding sites, one in the helicase core domain and the other in the carboxyl-terminal domain that recognizes 23S rRNA and interacts specifically with hairpin 92 of the PTC.

Amino Acid Motifs↗

Oxidation of alkylarenes to the corresponding acids using aqueous potassium permanganate by hydrodynamic cavitation.

Oxidation of toluene using aqueous potassium permanganate was studied under heterogeneous condition in the presence of hydrodynamic cavitation and compared with the results of the reaction under acoustic cavitation. Various parameters, such as quantity of potassium permanganate, toluene to aqueous phase ratio, reaction time and cavitation parameters such as orifice plate, and pump discharge pressure were optimized. The reaction was found to be considerably accelerated at ambient temperature in the presence of cavitation. On comparison, it was found that when 1 kJ of energy was passed to the reaction mixture in the case of acoustic cavitation, the product obtained was 4.63 x 10(-6) mol, whereas when 1 kJ of energy was passed to the reaction mixture in the case of hydrodynamic cavitation the product obtained was 2.70 x 10(-5) mol. Hence, about six times more product would be obtained in the case of hydrodynamic cavitation than in the case of acoustic cavitation at same energy dissipation. It has been observed that further optimization is possible.

Journal Article↗

Application of salicylic acid dosimetry to evaluate hydrodynamic cavitation as an advanced oxidation process.

The generation of OH* radicals inside hydrodynamic cavitation bubbles was monitored using a salicylic acid dosimeter. The reaction of this scavenger with OH* produces 2,5-dihydroxybenzoic acid (2,5-DHB) and, to a lesser degree, 2,3-DHB. The former, is a specific reaction product that can be determined with a very high sensitivity using HPLC-IF. This method has been applied to study the influence of the flow-rate and the solution pH for a given cavitation chamber geometry. The salicylic dosimetry has proven especially suitable for the characteristic time scales of hydrodynamic cavitation (higher than those of ultrasonic cavitation), which usually gives rise to recombination of radicals before they can reach the liquid-phase. Working at low pH the hydrophobic salicylic acid migrates to the gas-liquid interface and reacts with the OH* radicals, increasing the trapping efficiency of the dosimeter. Hydrodynamic cavitation works as a very low frequency sonochemical reactor, and therefore its potential as an Advanced Oxidation Process might be limited to reactions at the gas-liquid interface and inner bubble (i.e. with volatiles and/or hydrophobic substances).

Journal Article↗

Superior hydrodynamics of a modified cavopulmonary connection for the Norwood operation.

BACKGROUND: In the Fontan circulation, energy consumption at the cavopulmonary connection is crucial. Our hypothesis was that a modification of the standard Norwood variant of cavopulmonary connection with an extended anastomosis would improve hydrodynamics. METHODS: The in vitro hydrodynamics of two different Perspex glass models resembling the Norwood variant of cavopulmonary connection (model I) and the modification (model II) were analyzed in a mock circulation at nonpulsatile flows of 2 to 5 L/min to simulate rest and exercise. The pulmonary flow split was varied to imitate varying lung resistances. Inferior-to-superior caval flow ratio and size of models were increased to simulate growth. RESULTS: The pulmonary flow was preferentially directed to the left lung in model I and was better balanced in model II. Power losses increased exponentially with total flow in both models and were markedly higher in model I. These differences were attenuated in the larger models. Anastomotic turbulences were larger in model I. Power losses in both models were relatively insensitive to changes in pulmonary flow split. CONCLUSIONS: The proposed modification of the Norwood variant of cavopulmonary connection seems to be hydrodynamically advantageous and warrants further evaluation.

Anastomosis, Surgical↗

Calculation of hydrodynamic properties of globular proteins from their atomic-level structure.

The solution properties, including hydrodynamic quantities and the radius of gyration, of globular proteins are calculated from their detailed, atomic-level structure, using bead-modeling methodologies described in our previous article (, Biophys. J. 76:3044-3057). We review how this goal has been pursued by other authors in the past. Our procedure starts from a list of atomic coordinates, from which we build a primary hydrodynamic model by replacing nonhydrogen atoms with spherical elements of some fixed radius. The resulting particle, consisting of overlapping spheres, is in turn represented by a shell model treated as described in our previous work. We have applied this procedure to a set of 13 proteins. For each protein, the atomic element radius is adjusted, to fit all of the hydrodynamic properties, taking values close to 3 A, with deviations that fall within the error of experimental data. Some differences are found in the atomic element radius found for each protein, which can be explained in terms of protein hydration. A computational shortcut makes the procedure feasible, even in personal computers. All of the model-building and calculations are carried out with a HYDROPRO public-domain computer program.

Computer Simulation↗

The conformation of serum albumin in solution: a combined phosphorescence depolarization-hydrodynamic modeling study.

There is a striking disparity between the heart-shaped structure of human serum albumin (HSA) observed in single crystals and the elongated ellipsoid model used for decades to interpret the protein solution hydrodynamics at neutral pH. These two contrasting views could be reconciled if the protein were flexible enough to change its conformation in solution from that found in the crystal. To investigate this possibility we recorded the rotational motions in real time of an erythrosin-bovine serum albumin complex (Er-BSA) over an extended time range, using phosphorescence depolarization techniques. These measurements are consistent with the absence of independent motions of large protein segments in solution, in the time range from nanoseconds to fractions of milliseconds, and give a single rotational correlation time phi(BSA, 1 cP, 20 degrees C) = 40 +/- 2 ns. In addition, we report a detailed analysis of the protein hydrodynamics based on two bead-modeling methods. In the first, BSA was modeled as a triangular prismatic shell with optimized dimensions of 84 x 84 x 84 x 31.5 A, whereas in the second, the atomic-level structure of HSA obtained from crystallographic data was used to build a much more refined rough-shell model. In both cases, the predicted and experimental rotational diffusion rate and other hydrodynamic parameters were in good agreement. Therefore, the overall conformation in neutral solution of BSA, as of HSA, should be rigid, in the sense indicated above, and very similar to the heart-shaped structure observed in HSA crystals.

Animals↗

Hydrodynamic forces applied on intercellular bonds, soluble molecules, and cell-surface receptors.

Cells and biomolecules exposed to blood circulation experience hydrodynamic forces that affect their function. We present a methodology to estimate fluid forces and force loading rates applied on cellular aggregates, cell-surface proteins, and soluble molecules. Low Reynolds-number hydrodynamic theory is employed. Selected results are presented for biological cases involving platelets, neutrophils, tumor cells, GpIb-like cell-surface receptors, and plasma von Willebrand factor (vWF)-like soluble proteins. Calculations reveal the following: 1), upon application of constant linear shear, cell aggregates and biomolecules experience time-varying forces due to their tumbling motion. 2), In comparison to neutrophil homotypic aggregates, the maximum force applied on neutrophil-platelet aggregates is approximately threefold lower. Thus, alterations in cell size may dramatically alter adhesion molecule requirement for efficient cell binding. Whereas peak forces on homotypic cell doublets are tensile, shear forces dominate in heterotypic doublets with radius ratio <0.3. 3), The peak forces on platelet GpIb and von Willebrand factor are of comparable magnitude. However, they are orders-of-magnitude lower than those applied on intercellular bonds. Charts are provided to rapidly evaluate the magnitude of hydrodynamic force and rotation time-period occurring in any given experiment. The calculation scheme may find application in studies of vascular biology and receptor biophysics.

Blood Platelets↗

Hydrodynamic model of temperature change in open ionic channels.

Most theories of open ionic channels ignore heat generated by current flow, but that heat is known to be significant when analogous currents flow in semiconductors, so a generalization of the Poisson-Nernst-Planck theory of channels, called the hydrodynamic model, is needed. The hydrodynamic theory is a combination of the Poisson and Euler field equations of electrostatics and fluid dynamics, conservation laws that describe diffusive and convective flow of mass, heat, and charge (i.e., current), and their coupling. That is to say, it is a kinetic theory of solute and solvent flow, allowing heat and current flow as well, taking into account density changes, temperature changes, and electrical potential gradients. We integrate the equations with an essentially nonoscillatory shock-capturing numerical scheme previously shown to be stable and accurate. Our calculations show that 1) a significant amount of electrical energy is exchanged with the permeating ions; 2) the local temperature of the ions rises some tens of degrees, and this temperature rise significantly alters for ionic flux in a channel 25 A long, such as gramicidin-A; and 3) a critical parameter, called the saturation velocity, determines whether ionic motion is overdamped (Poisson-Nernst-Planck theory), is an intermediate regime (called the adiabatic approximation in semiconductor theory), or is altogether unrestricted (requiring the full hydrodynamic model). It seems that significant temperature changes are likely to accompany current flow in the open ionic channel.

Electrochemistry↗

Theory for the hydrodynamic and electrophoretic stretch of tethered B-DNA.

We have developed a theory for the extension and force of B-DNA tethered at a fixed point in a uniform hydrodynamic flow or in a uniform applied electric field. The chain tethered in an electric field is considered to be subject to free electrophoresis compensated by free sedimentation in the opposite direction. This allows the use of results of free electrophoresis for including the effects of small ions. The force on the chain is derived for a sequence of ellipsoidal segments, each twice the persistence length of the wormlike chain. Hydrodynamic interaction between these segments is based on the long-range limit of flow around the prolate ellipsoids, as derived from equivalent Stokes spheres. The chain extension is derived by applying the entropic elasticity relation of Marko and Siggia (1995 Macromolecules. 28:8759-8770) to each segment for polymer chains under constant tension. We justify this procedure by comparing with extension results based on the Boltzmann averaged orientation of straight, freely jointed segments. Predicted results agree well with recent extension-flow experiments by Perkins et al., 1995. Science. 258:83-87, and with electrophoretic stretch experiments by Smith and Bendich (1990 Biopolymers. 29:1167-1173) on fluorescently stained B-DNA. We find that the equivalence of hydrodynamic and electrophoretic stretch, proposed by Long et al. (1996 Phys. Rev. Lett. 76:3858-3861; 1996 Biopolymers 39:755-759), is valid only for very small chain deformations, but not in general.

Biophysical Phenomena↗

Experimental observations of the hydrodynamic behavior of solvent systems in high-speed counter-current chromatography. II. Phase distribution diagrams for helical and spiral columns.

Extensive studies have been performed on hydrodynamic distribution of two immiscible solvent phases in helical and spiral columns by means of coil planet centrifugation. Under the Scheme IV [see Fig. 1 in Y. Ito, J. Chromatogr., 301 (1984) 377] planetary motion, the two phases display unilateral distribution in the coil, i.e., one phase travels toward the head and the other phase toward the tail. This hydrodynamic trend is sensitively affected by various factors such as physical properties of the solvent system, helical diameter of the column, revolutional radius, etc. Phase distribution diagrams obtained from a set of conventional two-phase solvent systems under various experimental conditions provide extremely useful information for both basic hydrodynamic studies and practical application in high-speed counter-current chromatography.

Chromatography, High Pressure Liquid↗