A cell-based constitutive relation for bio-artificial tissues.
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By using a combination of continuum and statistical mechanics we derive an integral constitutive relation for bio-artificial tissue models consisting of a monodisperse population of cells in a uniform collagenous matrix. This constitutive relation quantitatively models the dependence of tissue stress on deformation history, and makes explicit the separate contribution of cells and matrix to the mechanical behavior of the composite tissue. Thus microscopic cell mechanical properties can be deduced via this theory from measurements of macroscopic tissue properties. A central feature of the constitutive relation is the appearance of "anisotropy tensors" that embody the effects of cell orientation on tissue mechanics. The theory assumes that the tissues are stable over the observation time, and does not in its present form allow for cell migration, reorientation, or internal remodeling. We have compared the predictions of the theory to uniaxial relaxation tests on fibroblast-populated collagen matrices (FPMs) and find that the experimental results generally support the theory and yield values of fibroblast contractile force and stiffness roughly an order of magnitude smaller than, and viscosity comparable to, the corresponding properties of active skeletal muscle. The method used here to derive the tissue constitutive equation permits more sophisticated cell models to be used in developing more accurate representations of tissue properties. (+info)
Anisotropic reentry in a perfused 2-dimensional layer of rabbit ventricular myocardium.
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BACKGROUND: Anisotropy creates nonuniformity in electrical propagation and may contribute to the occurrence of unidirectional conduction block and reentry. We describe the characteristics of reentrant tachycardia in a 2D layer of anisotropic ventricular myocardium. METHODS AND RESULTS: A Langendorff-perfused epicardial sheet (1.0+/-0.4 mm, n=35) was created by freezing the intramural layers of the rabbit left ventricle. Epicardial activation maps were constructed by use of different high-resolution mapping arrays connected to a mapping system. In 5 experiments, monophasic action potentials were recorded. In the intact left ventricle, no arrhythmias except VF could be induced. After freezing, programmed electrical stimulation or rapid pacing led to the induction of sustained VT (cycle length 130+/-11 ms). VT was caused by reentry around a functional line of block oriented parallel to the epicardial fiber direction. Action potential recordings demonstrated that the central line of block was kept refractory by electrotonic currents generated by the depolarization waves propagating at either side of the line of block. At the pivot points of the line of block, the pronounced curvature of the turning wave and abrupt loading changes created an excitable gap of 30 ms in the reentrant pathway. CONCLUSIONS: In uniform anisotropic myocardium, reentry around a functional Z-shaped line of block may occur. The core of the circuit is kept refractory by electrotonic currents. The pronounced wave-front curvature and abrupt loading changes at the pivot points cause local conduction delay and create a small excitable gap. (+info)
Diffusion-weighted MR imaging of the normal human spinal cord in vivo.
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BACKGROUND AND PURPOSE: Diffusion-weighted imaging is a robust technique for evaluation of a variety of neurologic diseases affecting the brain, and might also have applications in the spinal cord. The purpose of this study was to determine the feasibility of obtaining in vivo diffusion-weighted images of the human spinal cord, to calculate normal apparent diffusion coefficient (ADC) values, and to assess cord anisotropy. METHODS: Fifteen healthy volunteers were imaged using a multi-shot, navigator-corrected, spin-echo, echo-planar pulse sequence. Axial images of the cervical spinal cord were obtained with diffusion gradients applied along three orthogonal axes (6 b values each), and ADC values were calculated for white and gray matter. RESULTS: With the diffusion gradients perpendicular to the orientation of the white matter tracts, spinal cord white matter was hyperintense to central gray matter at all b values. This was also the case at low b values with the diffusion gradients parallel to the white matter tracts; however, at higher b values, the relative signal intensity of gray and white matter reversed. With the diffusion gradients perpendicular to spinal cord, mean ADC values ranged from 0.40 to 0.57 x 10(-3) mm2/s for white and gray matter. With the diffusion gradients parallel to the white matter tracts, calculated ADC values were significantly higher. There was a statistically significant difference between the ADCs of white versus gray matter with all three gradient directions. Strong diffusional anisotropy was observed in spinal cord white matter. CONCLUSION: Small field-of-view diffusion-weighted images of the human spinal cord can be acquired in vivo with reasonable scan times. Diffusion within spinal cord white matter is highly anisotropic. (+info)
Diffusion tensor MR imaging of the brain: effect of diffusion weighting on trace and anisotropy measurements.
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BACKGROUND AND PURPOSE: In human brain, the relationship between MR signal and b value is complicated by cerebral perfusion, restricted diffusion, anisotropy, cellular membrane permeability, and active cellular transport of water molecules. Our purpose was to evaluate the effect of the number and strength of diffusion-sensitizing gradients on measured isotropic apparent diffusion coefficients (ADCi), fractional anisotropy (FA), and their respective SD in different anatomic locations of the brain. METHODS: Quantitative apparent diffusion coefficients and diffusion anisotropy brain maps were obtained from 10 healthy volunteers by using six different levels of diffusion weighting (b0 = 0, bl = 160, b2 = 320, b3 = 480, b4 = 640, and b5 = 800 s/mm2), applied sequentially in six different directions (Gxx, Gyy, Gzz, Gxy, Gxz, Gyz) and coupled to a single-shot spin-echo echo-planar (2,045/115 [TR/TE]) MR imaging technique. ADCi, FA, eigenvalues (lambda1, lambda2, lamdba3)1 of the principal eigenvectors, and their respective SD were measured from seven different anatomic locations in the brain. Repeated measures analysis of variance was used to evaluate for the existence of significant differences in the average and SD of the calculated ADCi and FA as a function of the number and strength of b values. When a difference existed, the Bonferroni t method was used for paired comparisons of the groups. RESULTS: The measured ADCi was affected by the number and strength of b values (P < .05). The SD of the ADCi was affected by the strength (P < .05) but not the number of b values (P > .05). The measured FA was unaffected by the number and strength of b values (P > .05). The SD was affected by the number and strength of b values (P < .05). CONCLUSION: The number and strength of b values do influence measures of diffusion and anisotropy. Attention to the choice of diffusion sensitization parameters is important in decisions regarding clinical feasibility (acquisition time) and normative measures. (+info)
Ambiguity in the perception of moving stimuli is resolved in favour of the cardinal axes.
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The aim of this study was to determine whether there is a link between the statistical properties of natural scenes and our perception of moving surfaces. Accordingly, we devised an ambiguous moving stimulus that could be perceived as moving in one of three directions of motion. The stimulus was a circular patch containing three square-wave drifting gratings. One grating was always either horizontal or vertical; the other two had component directions of drift at 120 degrees to the first (and to each other), producing four possible stimulus geometries. These were presented in a pseudorandom sequence. In brief presentations, subjects always perceived two of the gratings to cohere and move as a pattern in one direction, and the third grating to move independently in the opposite direction (its component direction). Although there were three equally plausible axes (one cardinal and two oblique) along which the coherent and independent motions could occur, subjects routinely saw motion along one of the cardinal axes. Thus, the visual system preferentially combines the two oblique gratings to form a pattern that drifts in the opposite direction to the cardinal grating. It was only when the contrast of one of the oblique gratings was changed that an oblique axis of motion was perceived. This perceptual anisotropy can be related to naturally occurring bias in the visual environment, notably the predominance of horizontal and vertical contours in our visual world. (+info)
A fluorescence study of the interaction and location of (+)-totarol, a diterpenoid bioactive molecule, in model membranes.
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(+)-Totarol, a diterpene extracted from Podocarpus totara, has been reported as a potent antioxidant and antibacterial agent. Although the molecular mechanism of action of this hydrophobic molecule remains unknown, recent work made in our laboratory strongly suggests that it could be lipid-mediated. Since (+)-totarol contains a phenolic ring, we have studied the intrinsic fluorescent properties of this molecule, i.e., quantum yield, lifetime, steady-state anisotropy and emission spectra, both in aqueous and in phospholipid phases, in order to obtain information on the interaction and location of (+)-totarol in biomembrane model systems. The phospholipid/water partition coefficient of (+)-totarol was found to be very high (K(p)=1.8x10(4)), suggesting that it incorporates very efficiently into membranes. In order to estimate the transverse location (degree of penetration) of the molecule in the fluid phase of DMPC model membranes, the spin labelled fatty acids 5-NS and 16-NS were used in differential quenching experiments. The results obtained show that (+)-totarol is located in the inner region of the membrane, far away from the phospholipid/water interface. Since (+)-totarol protects against oxidative stress, its interaction with an unsaturated fatty acid, trans-parinaric acid, was studied using fluorescence resonance energy transfer. No significant interactions were observed, molecules of trans-parinaric acid distributing themselves randomly amongst those of (+)-totarol in the phospholipid membrane. (+info)
Equilibrium binding of single-stranded DNA to the secondary DNA binding site of the bacterial recombinase RecA.
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The bacterial recombinase RecA forms a nucleoprotein filament in vitro with single-stranded DNA (ssDNA) at its primary DNA binding site, site I. This filament has a second site, site II, which binds ssDNA and double-stranded DNA. We have investigated the binding of ssDNA to the RecA protein in the presence of adenosine 5'-O-(thiotriphosphate) cofactor using fluorescence anisotropy. The RecA protein carried out DNA strand exchange with a 5'-fluorescein-labeled 32-mer oligonucleotide. The anisotropy signal was shown to measure oligonucleotide binding to RecA, and the relationship between signal and binding density was determined. Binding of ssDNA to site I of RecA was stable at high NaCl concentrations. Binding to site II could be described by a simple two-state equilibrium, K = 4.5 +/- 1.5 x 10(5) m(-1) (37 degrees C, 150 mm NaCl, pH 7.4). The reaction was enthalpy-driven and entropy-opposed. It depended on salt concentration and was sensitive to the type of monovalent anion, suggesting that anion-dependent protein conformations contribute to ssDNA binding at site II. (+info)
Solution structure of hpTX2, a toxin from Heteropoda venatoria spider that blocks Kv4.2 potassium channel.
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HpTX2 is a toxin from the venom of Heteropoda venatoria spider that has been demonstrated to bind on Kv4.2 potassium channel. We have determined the solution structure of recombinant HpTX2 by use of conventional two-dimensional NMR techniques followed by distance-geometry and molecular dynamics. The calculated structure belongs to the Inhibitory Cystin Knot structural family that consists in a compact disulfide-bonded core, from which four loops emerge. A poorly defined two-stranded antiparallel beta-sheet (residues 20-23 and 25-28) is detected. Analysis of the electrostatic charge anisotropy allows us to propose a functional map of HpTX2 different from the one described for kappa-conotoxin PVIIA, but strongly related to the one of charybdotoxin. The orientation of the dipole moment of HpTX2 emerges through K27 which could therefore be the critical lysine residue. Close to this lysine are a second basic residue, R23, an aromatic cluster (F7, W25, W30) and an hydrophobic side chain (L24). The high density in aromatic side chains of the putative functional surface as well as the lack of an asparagine is proposed to be the structural basis of the specificity of HpTX2 toward Kv4.2 channel. (+info)