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PubMed1985

Hydrodynamic characterizations of estrogen receptors complexed with [3H]-4-hydroxytamoxifen: evidence in support of contrasting receptor transitions mediated by different ligands.

Size-exclusion high-performance liquid chromatography was used to characterize the hydrodynamic molecular properties of estrogen receptors complexed with estradiol and the antiestrogen 4-hydroxytamoxifen. Cytoplasmic estrogen receptors complexed with [3H]-4-hydroxytamoxifen did not undergo reductions in hydrodynamic size after exposure to KCl or urea. Nuclear receptors complexed with 4-hydroxytamoxifen eluted as hydrodynamically larger molecules than nuclear receptors complexed with estradiol. Because identical hydrodynamic characterizations were obtained with the covalent ligand [3H]tamoxifen aziridine, these differences in chromatographic behavior are due to differences in ligand-mediated receptor properties and are not the result of ligand dissociation. When estrogen receptors, complexed with either [3H]estradiol or [3H]-4-hydroxytamoxifen, were exposed to trypsin, the receptors complexed with 4-hydroxytamoxifen eluted as larger hydrodynamic forms than receptors complexed with estradiol. These observations are interpreted to indicate that estradiol and 4-hydroxytamoxifen mediate contrasting transitions in the molecular orientation of estrogen receptors. The consequences of the transitions mediated by 4-hydroxytamoxifen appear to be that intermolecular associations become difficult to disrupt with KCl or urea and that the accessibility of trypsin-sensitive proteolytic sites becomes altered. Chromatin fractionation using DNase I and hypotonic Mg2+ solubilization identified a chromatin region that was less readily penetrated by receptors complexed with 4-hydroxytamoxifen than receptors complexed with estradiol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Study of physical and biological factors involved in the disruption of E. coli by hydrodynamic cavitation.

Hydrodynamic cavitation results in flow restriction in a flow system causing rapid pressure fluctuations and significant fluid forces. These can be harnessed to mediate microbial cell damage. Hydrodynamic cavitation was studied for the partial disruption of E. coli and selective release of specific proteins relative to the total soluble protein. The effects of the cavitation number, the number of passes, and the specific growth rate of E. coli on the release of periplasmic and cytoplasmic proteins were studied. At the optimum cavitation number of 0.17 for this experimental configuration, 48% of the total soluble protein, 88% of acid phosphatase, and 67% of beta-galactosidase were released by hydrodynamic cavitation in comparison with the maximum release attained using multiple passes through the French Press. The higher release of the acid phosphatase over the total soluble protein suggested preferred release of periplasmic compounds. This was supported by SDS-PAGE analysis. The absence of micronization of cell material resulting in the potential for ease of solid-liquid separation downstream of the cell disruption operation was confirmed by TEM microscopy. E. coli cells cultivated at a higher specific growth rate (0.36 h(-1)) were more easily disrupted than slower grown cells (0.11 h(-1)). The specific activity of the enzyme of interest released by hydrodynamic cavitation, defined as the units of enzyme in solution per milligram of total soluble protein, was greater than that obtained on release by the French Press, high-pressure homogenization, osmotic shock, and EDTA treatment. The selectivity offered indicates the potential of enzyme release by hydrodynamic cavitation to ease the purification in the subsequent downstream processing.

Acid Phosphatase↗
PubMed2005

Role of hydrodynamic drag on microsphere deposition and re-entrainment in porous media under unfavorable conditions.

Deposition and re-entrainment of 1.1 microm microspheres were examined in packed glass beads and quartz sand under both favorable and unfavorable conditions for deposition. Experiments were performed at environmentally relevant ionic strengths and flow rates in the absence of solution chemistry and flow perturbations. Numerical simulations of experimental data were performed using kinetic rate coefficients to represent deposition and re-entrainment dynamics. Deposition rate coefficients increased with increasing flow rate under favorable deposition conditions (in the absence of colloid-grain surface electrostatic repulsion), consistent with expected trends from filtration theory. In contrast, under unfavorable deposition conditions (where significant colloid-grain surface electrostatic repulsion exists), the deposition rate coefficients decreased with increasing flow rate, suggesting a mitigating effect of hydrodynamic drag on deposition. Furthermore, the re-entrainment rate was negligible under favorable conditions but was significant under unfavorable conditions and increased with increasing flow rate, demonstrating that hydrodynamic drag drove re-entrainment under unfavorable conditions. The drag torque resulting from hydrodynamic drag was found to be 1 order of magnitude or more lower than the adhesive torque based on pull-off forces from atomic force microscopy measurements. This result indicates that hydrodynamic drag was insufficient to drive re-entrainment of microspheres that were associated with the grain surface via the primary energy minimum and suggests that hydrodynamic drag drove re-entrainment of secondary-minimum-associated microspheres.

Biomechanical Phenomena↗
PubMed2005

Strong effect of hydrodynamic coupling on the electric dichroism of bent rods.

The effect of hydrodynamic coupling on the spatial orientation of rigid bent rods in electric fields has been analyzed by Brownian dynamics simulations. Bead models for smoothly bent rods were constructed with dimensions of DNA double helices, and established simulation procedures were used to calculate their diffusion tensor, including the translational-rotational coupling tensor. The electric and optical parameters were assigned on the basis of known properties of double helices. Brownian dynamics simulations of the orientation of these models in electric fields showed that both transients and amplitudes of the calculated dichroism are very strongly dependent on translational-rotational coupling over a wide range of electric field strengths. For example, the stationary dichroism of a smoothly bent 179 bp DNA fragment calculated at low field strengths is positive in the presence and negative in the absence of hydrodynamic coupling. The transients are converted from a biphasic to a monophasic shape, when hydrodynamic coupling is turned off. The large changes resulting from hydrodynamic coupling were controlled by calculations based on analytical expressions derived for electrooptical response curves in the limit of low electric field strengths; the results obtained by this independent approach are in very satisfactory agreement with our Brownian dynamics simulations. The effect is strongly dependent on the electric dipole and on its direction. In the absence of any dipole the coupling effect was not observed. The coupling effect increases with the size of the bent rods. Because most macromolecular structures are known to have induced and/or permanent dipole moments, large effects of hydrodynamic coupling on both the amplitudes and the transients of the electric dichroism/birefringence must be expected in general for structures with nonsymmetric shape.

Journal Article↗
PubMed2002

Influence of hydrodynamics and particle size on the absorption of felodipine in labradors.

PURPOSE: To study the influence of GI hydrodynamics and drug particle size on felodipine absorption in the dog. METHODS: Labradors fistulated at midjejunum were used to selectively study the influence of hydrodynamics and particle size on the in vivo dissolution and absorption of the poorly soluble, lipophilic drug felodipine. A combination of infusion and oral administration of either normal saline or a 5% glucose solution was used to maintain "fasted" and establish "fed" state motility patterns, respectively. The absorption characteristics of both a micronized (8 microm) and a coarse fraction (125 micom) of felodipine were subsequently studied under these two motility patterns. RESULTS: A reduction in particle size led up to an approximate 22-fold increase in maximum plasma concentration and up to an approximate 14-fold increase in area under the curve, with a commensurate decrease in the time at which the maximum plasma concentration occurred. Although the absorption of felodipine from the solution and micronized suspension was not influenced by a change in the hydrodynamics, felodipine was absorbed from the coarse suspension almost twice as well in the "fed" state as under "fasted" conditions. CONCLUSIONS: Absorption from coarse suspensions of felodipine was sensitive to luminal hydrodynamics, whereas micronized suspensions were not. However, the particle size seems to have a much more important influence on the bioavailability of felodipine than the hydrodynamics per se.

Administration, Oral↗
PubMed2005

Hydrodynamic liver gene transfer mechanism involves transient sinusoidal blood stasis and massive hepatocyte endocytic vesicles.

The present study contributes to clarify the mechanism underlying the high efficacy of hepatocyte gene transfer mediated by hydrodynamic injection. Gene transfer experiments were performed employing the hAAT gene, and the efficacy and differential identification in mouse plasma of human transgene versus mouse gene was assessed by ELISA and proteomic procedures, respectively. By applying different experimental strategies such as cumulative dose-response efficacy, hemodynamic changes reflected by venous pressures, intravital microscopy, and morphological changes established by transmission electron microscopy, we found that: (a) cumulative multiple doses of transgene by hydrodynamic injection are efficient and well tolerated, resulting in therapeutic plasma levels of hAAT; (b) hydrodynamic injection mediates a transient inversion of intrahepatic blood flow, with circulatory stasis for a few minutes mainly in pericentral vein sinusoids; (c) transmission electron microscopy shows hydrodynamic injection to promote massive megafluid endocytic vesicles among hepatocytes around the central vein but not in hepatocytes around the periportal vein. We suggest that the mechanism of hydrodynamic liver gene transfer involves transient inversion of intrahepatic flow, sinusoidal blood stasis, and massive fluid endocytic vesicles in pericentral vein hepatocytes.

Animals↗
PubMed2006

Structural impact of hydrodynamic injection on mouse liver.

The impact of hydrodynamic injection on liver structure was evaluated in mice using various microscopic techniques. Upon hydrodynamic injection of approximately 9% of body weight by volume, the liver rapidly expanded, reaching maximal size at the end of the injection and returned to its original size in 30 min. Histological analysis revealed a swollen appearance in the peri-central region of the liver where delivery of genes and fluorescence-labeled markers was observed. Scanning and transmission electron microscopy showed enlargement and rupture of endothelium that in about 24-48 h regains its morphology and normal function as a barrier against infection by adenovirus viral particles. At the cellular level in hydrodynamically treated animals, four types of hepatocytes were seen: cells with normal appearance; cells with enriched vesicles in the cytoplasm; cells with lightly stained cytosol; and cells with significant dilution of the cytoplasm. In addition, red blood cells and platelets were observed in the space of Disse and even inside hepatocytes. Vesicle formation is triggered by hydrodynamic injection and resembles the process of macropinocytosis. These results, whereas confirming the physical nature of hydrodynamic delivery, are important for a better understanding of this efficient method for intrahepatic gene and small interfering RNA delivery.

Adenoviridae↗
PubMed2005

Brownian dynamics simulations of a flexible polymer chain which includes continuous resistance and multibody hydrodynamic interactions.

Using methods adapted from the simulation of suspension dynamics, we have developed a Brownian dynamics algorithm with multibody hydrodynamic interactions for simulating the dynamics of polymer molecules. The polymer molecule is modeled as a chain composed of a series of inextensible, rigid rods with constraints at each joint to ensure continuity of the chain. The linear and rotational velocities of each segment of the polymer chain are described by the slender-body theory of Batchelor [J. Fluid Mech. 44, 419 (1970)]. To include hydrodynamic interactions between the segments of the chain, the line distribution of forces on each segment is approximated by making a Legendre polynomial expansion of the disturbance velocity on the segment, where the first two terms of the expansion are retained in the calculation. Thus, the resulting linear force distribution is specified by a center of mass force, couple, and stresslet on each segment. This method for calculating the hydrodynamic interactions has been successfully used to simulate the dynamics of noncolloidal suspensions of rigid fibers [O. G. Harlen, R. R. Sundararajakumar, and D. L. Koch, J. Fluid Mech. 388, 355 (1999); J. E. Butler and E. S. G. Shaqfeh, J. Fluid Mech. 468, 204 (2002)]. The longest relaxation time and center of mass diffusivity are among the quantities calculated with the simulation technique. Comparisons are made for different levels of approximation of the hydrodynamic interactions, including multibody interactions, two-body interactions, and the "freely draining" case with no interactions. For the short polymer chains studied in this paper, the results indicate a difference in the apparent scaling of diffusivity with polymer length for the multibody versus two-body level of approximation for the hydrodynamic interactions.

Journal Article↗
PubMed2006

Electro-osmosis at inhomogeneous charged surfaces: hydrodynamic versus electric friction.

Electrokinetic methods are efficient in probing the electrostatic surface properties of charged systems. However, anomalies observed in experiments indicate that the classical electrokinetic theory should be reconsidered. Using Green's function methods and hydrodynamic simulations, we investigate electro-osmosis driven by electric-field-induced ion motion near a charged planar substrate with smooth or rough boundary. First, a reformulation of electro-osmotic theory for planar charged surfaces employing Green's functions shows that the Helmholtz-Smoluchowski (HS) relation between electrostatic potential and solvent velocity is exact for smooth surfaces, even in the presence of ion correlations. Deviations from HS theory are caused by combined hydrodynamic and electric surface friction, as our hydrodynamic simulations of ions at smooth and corrugated charged surfaces in lateral electric fields demonstrate. Within the simulations, hydrodynamic interactions are treated in the continuum limit and the presence of a no-slip boundary condition at the surface is taken into account. While electrofriction is relevant in highly charged system and/or for multivalent ions, hydrodynamic friction is dominant in systems with moderate surface charge density and/or low ionic valency. We also derive the effective electrokinetic surface charge from the electro-osmotic solvent profiles, which is substantially reduced when compared with the bare value and shows qualitative agreement with the experimental tendency.

Journal Article↗
PubMed2002

Calculation of hydrodynamic properties of small nucleic acids from their atomic structure.

Hydrodynamic properties (translational diffusion, sedimentation coefficients and correlation times) of short B-DNA oligonucleotides are calculated from the atomic-level structure using a bead modeling procedure in which each non-hydrogen atom is represented by a bead. Using available experimental data of hydrodynamic properties for several oligonucleotides, the best fit for the hydrodynamic radius of the atoms is found to be approximately 2.8 A. Using this value, the predictions for the properties corresponding to translational motion and end-over-end rotation are accurate to within a few percent error. Analysis of NMR correlation times requires accounting for the internal flexibility of the double helix, and allows an estimation of approximately 0.85 for the Lipari-Szabo generalized order parameter. Also, the degree of hydration can be determined from hydrodynamics, with a result of approximately 0.3 g (water)/g (DNA). These numerical results are quite similar to those found for globular proteins. If the hydrodynamic model for the short DNA is simply a cylindrical rod, the predictions for overall translation and rotation are slightly worse, but the NMR correlation times and the degree of hydration, which depend more on the cross-sectional structure, are more severely affected.

Base Sequence↗
PubMed2004

Influence of hydrodynamics on many-particle diffusion in 2D colloidal suspensions.

We study many-particle diffusion in 2D colloidal suspensions with full hydrodynamic interactions through a novel mesoscopic simulation technique. We focus on the behaviour of the effective scaled tracer and collective-diffusion coefficients DT(rho)/D0 and DC(rho)/D0 respectively, where D0 is the single-particle diffusion coefficient, as a function of the density of the colloids rho. At low Schmidt numbers Sc - 1, we find that hydrodynamics has essentially no effect on the behaviour of DT (rho)/D0. At larger Sc, DT (rho)/D0 seems to be enhanced at all densities, although the differences compared to the case without hydrodynamics are rather minor. The collective-diffusion coefficient, on the other hand, is much more strongly coupled to hydrodynamical conservation laws and is distinctly different from the purely dissipative case without hydrodynamic interactions.

Colloids↗
PubMed2005

Integration of hydrodynamic and odorant inputs by local interneurons of the crayfish deutocerebrum.

Intracellular electrodes were used to record from local interneurons in the olfactory lobes of the midbrain in the crayfish Procambarus clarkii. Cells that resembled previously studied central targets of olfactory receptor neurons on the lateral antennular flagellum were specifically examined for their responses to hydrodynamic stimuli. Initiation of water movement past the antennular flagellum, confined within an olfactometer, evoked a triphasic excitatory-inhibitory-excitatory postsynaptic potential lasting up to 2 s that generated spikes on depolarizing phases of the response sequence. Odorant pulses seamlessly imbedded in the water pulse past the antennule evoked purely excitatory, dose-dependent postsynaptic responses and associated spike trains. The latency of the initial phase of the response to water was approximately half as long as the latency of the response to odorant, suggesting that different afferent pathways are involved in responses to hydrodynamic and odorant stimuli, respectively. In some olfactory lobe interneurons that resembled previously described cells classified as Type I, conjoint stimulation of fluid onset and odorant evoked responses that were twice the amplitude of the summed response to either hydrodynamic or odorant stimulation alone, suggesting that the olfactory responses were potentiated by hydrodynamic input. Individuals of at least one other class of first-order interneuron that responded to both hydrodynamic and odorant stimulation were occasionally recorded from. These results indicate that multimodal integration of chemical and mechanical information occurs at the level of first-order sensory interneurons in the crayfish brain.

Animals↗
PubMedDate not supplied

[Effect of additional loading on kinematics and hydrodynamics of dolphins].

The results of comparative measurements of hydrodynamic resistance of a solid model of dolphin and of additional load imitating an autonomous apparatus placed on the dolphin body are presented. The loading included cylinders (with rounded anterior and posterior ends) and spheres. They were fixed on the dorsal fin of the dolphin model in such a way that the interrelationship between the load and the model by a water stream minimal. In several experiments, a thick ring (dr = 10 mm) was fixed in the region of the dolphin respiratory opening to reach the turbulence of the boundary layer. The dolphin model with loads was towed in a speed range between 1.0 and 4.5 m/s. It was shown that the hydrodynamic resistance of additional loads is comparable with the resistance of the solid dolphin model. For the entire spectrum of loading in the preset range of towing speeds, the coefficient of hydrodynamic resistance of loading (according to the area of the wet body surface) was from 10 to 145% of the coefficient of the dolphin model resistance. It was assumed that the results of the measurements of kinematics and hydrodynamic characteristics of a freely-swimming dolphin can substantially differ from the results obtained on a dolphin carrying devices (at restricted power of the animal). Similar problems can also arise if the animals will carry devices with a higher hydrodynamic resistance.

Animals↗
PubMedDate not supplied

The effect of hydrodynamic shear on 3D engineered chondrocyte systems subject to direct perfusion.

Bioreactors allowing direct-perfusion of culture medium through tissue-engineered constructs may overcome diffusion limitations associated with static culturing, and may provide flow-mediated mechanical stimuli. The hydrodynamic stress imposed on cells within scaffolds is directly dependent on scaffold microstructure and on bioreactor configuration. Aim of this study is to investigate optimal shear stress ranges and to quantitatively predict the levels of hydrodynamic shear imposed to cells during the experiments. Bovine articular chondrocytes were seeded on polyestherurethane foams and cultured for 2 weeks in a direct perfusion bioreactor designed to impose 4 different values of shear level at a single flow rate (0.5 ml/min). Computational fluid dynamics (CFD) simulations were carried out on reconstructions of the scaffold obtained from micro-computed tomography images. Biochemistry analyses for DNA and sGAG were performed, along with electron microscopy. The hydrodynamic shear induced on cells within constructs, as estimated by CFD simulations, ranged from 4.6 to 56 mPa. This 12-fold increase in the level of applied shear stress determined a 1.7-fold increase in the mean content in DNA and a 2.9-fold increase in the mean content in sGAG. In contrast, the mean sGAG/DNA ratio showed a tendency to decrease for increasing shear levels. Our results suggest that the optimal condition to favour sGAG synthesis in engineered constructs, at least at the beginning of culture, is direct perfusion at the lowest level of hydrodynamic shear. In conclusion, the presented results represent a first attempt to quantitatively correlate the imposed hydrodynamic shear level and the invoked biosynthetic response in 3D engineered chondrocyte systems.

Animals↗
PubMed1996

Hydrodynamic thrombectomy of haemodialysis grafts and fistulae: results of 51 procedures.

PURPOSE: To describe efficacy of hydrodynamic thrombectomy for occluded dialysis native and graft fistulae in 51 instances. MATERIALS AND METHODS: Fifty-one hydrodynamic thrombectomies of 34 native and graft a-v fistulae were performed. There were 32 thrombectomies in PTFE grafts and 19 procedures in native Brescia-Cimino fistulae. Multiple thrombectomies were performed in 11 of 34 fistulae. The estimated occlusion time was 36.4+/-22 h. The length of the occluded segment ranged from 2 to 50 cm (mean 28.8 cm). In all cases, a 7 F hydrodynamic thrombectomy catheter was used. Double-cannulation technique was used for graft fistulae, single-cannulation for native fistulae. Additionally, balloon dilatation was performed in all 51 cases, stenting in six, and aspiration thrombectomy in two cases. RESULTS: Arterialized flow was re-established by hydrodynamic thrombectomy and PTA in 43 of 51 cases (84%). By additional use of other techniques, technical success improved to 46 of 51 procedures (90%). Early re-thrombosis occurred in six cases within 24 h of thrombectomy (11%). Clinical success was achieved in 39 of 46 technically successful cases (85%). Cumulative patency was calculated at 63% after 1 week, 57% after 1 month, 48% after 3 months, 37% after 6 months, and 32% after 12 months. Patency of native fistulae after thrombectomy was better than patency of grafts. CONCLUSIONS: Hydrodynamic thrombectomy is an effective percutaneous technique for declotting haemodialysis fistulae and grafts recently thrombosed.

Adult↗
PubMed1995

[Hydrodynamics of syringomyelia].

Abnormality of CSF hydrodynamics is an important factor of the pathogenesis of syringomyelia. Recent development of cine-MRI visualized real time CSF movement in the syrinx or subarachnoid space. In the present study, twenty cases of syringomyelia associated with Chiari I malformation, 10 cases of syringomyelia associated with spina bifida aperta, and 10 cases of syringomyelia associated with spina bifida occulta were studied with MRI and cine-MRI. The result demonstrated that hydrodynamics of these three groups were different to each other. Chiari I malformation showed CSF hydrodynamic abnormality at the foramen magnum and marked flow void in the syrinx. On the other hand, myelomeningocele showed hydrocephalus and less prominent flow void in the syrinx. Spinal lipoma which is the majority of spina bifida occulta showed no abnormality of CSF hydrodynamics. In conclusion, the selection of treatments should be individualized depending on the hydrodynamical abnormality of each patient.

Arnold-Chiari Malformation↗
PubMedDate not supplied

[Study of hydrodynamic characteristics of artificial heart valves by photochromic imaging].

The paper deals with the hydrodynamics of artificial heart valves (AHVs) used in clinical practice. It reviews and analyzes the studies of AHV hydrodynamics, as well as the hydrodynamic beds which stimulate physiological flow through the valve. Photochronic imaging (PCI) is proposed for examination of AHV hydrodynamic characteristics under model physiological flow. The hydrodynamics of different AHVs was tested on the beds simulating blood flow through AHVs by employing PCI. PCI involved preparation of model photochronic solution that simulates blood, colour labels by using laser radiation. In the model photochronic solution, 10(-6)-10(-9)-sec laser radiation gave rise to linear colour labels whose movement was recorded by a speed camera in the flow behind the valve. The profiles of speed behind the valves, the dimensions of congestive areas, the positions of flow detachment and regurgitation flow were calculated by a speed shooting in different periods of valvular performance. PCI defined congestive areas behind the valves, the areas of closed circulations, the sizes of reversing flow areas and examined the time course of flow behind the valves as a whole. The paper is of interest for AHV designers and cardiac surgeons who apply various AHVs.

Color↗
PubMed2005

Effect of background electrolyte on the estimation of protein hydrodynamic radius and net charge through capillary zone electrophoresis.

Two physicochemical models are proposed for the estimation of both hydrodynamic radius and net charge of a protein when the capillary zone electrophoretic mobility at a given protocol, the set of pK of charged amino acids, and basic data from Protein Data Bank are available. These models also provide a rationale to interpret appropriately the effects of solvent properties on protein hydrodynamic radius and net charge. To illustrate the numerical predictions of these models, experimental data of electrophoretic mobility available in the literature for well-defined protocols are used. Five proteins are considered: lysozyme, staphylococcal nuclease, human carbonic anhydrase, bovine carbonic anhydrase, and human serum albumin. Numerical predictions of protein net charges through these models compare well with the results reported in the literature, including those found asymptotically through protein charge ladder techniques. Model calculations indicate that the hydrodynamic radius is sensitive to changes of the protein net charge and hence it cannot be assumed constant in general. Also, several limitations associated with models for estimating protein net charge and hydrodynamic radius from protein structure, amino acid sequence, and experimental electrophoretic mobility are provided and discussed. These conclusions also show clear requirements for further research.

Electrolytes↗