Shaping surface acoustic waves for cardiac tissue engineering

The heart is a non-regenerating organ that gradually suffers a loss of cardiac cells and functionality. Given the scarcity of organ donors and complications in existing medical implantation solutions, it is desired to engineer a three-dimensional architecture to successfully control the cardiac cells in vitro and yield true myocardial structures similar to native heart. This thesis investigates the synthesis of a biocompatible gelatin methacrylate hydrogel to promote growth of cardiac cells using biotechnology methodology: surface acoustic waves, to create cell sheets. Firstly, the synthesis of a photo-crosslinkable gelatin methacrylate (GelMA) hydrogel was investigated with different degree of methacrylation concentration. The porous matrix of the hydrogel should be biocompatible, allow cell-cell interaction and promote cell adhesion for growth through the porous network of matrix. The rheological properties, such as polymer concentration, ultraviolet exposure time, viscosity, elasticity and swelling characteristics of the hydrogel were investigated. In tissue engineering hydrogels have been used for embedding cells to mimic native microenvironments while controlling the mechanical properties. Gelatin methacrylate hydrogels have the advantage of allowing such control of mechanical properties in addition to easy compatibility with Lab-on-a-chip methodologies. Secondly in this thesis, standing surface acoustic waves were used to control the degree of movement of cells in the hydrogel and produce three-dimensional engineered scaffolds to investigate in-vitro studies of cardiac muscle electrophysiology and cardiac tissue engineering therapies for myocardial infarction. The acoustic waves were characterized on a piezoelectric substrate, lithium niobate that was micro-fabricated with slanted-finger interdigitated transducers for to generate waves at multiple wavelengths. This characterization successfully created three-dimensional micro-patterning of cells in the constructs through means of one- and two-dimensional non-invasive forces. The micro-patterning was controlled by tuning different input frequencies that allowed manipulation of the cells spatially without any pre- treatment of cells, hydrogel or substrate. This resulted in a synchronous heartbeat being produced in the hydrogel construct. To complement these mechanical forces, work in dielectrophoresis was conducted centred on a method to pattern micro-particles. Although manipulation of particles were shown, difficulties were encountered concerning the close proximity of particles and hydrogel to the microfabricated electrode arrays, dependence on conductivity of hydrogel and difficult manoeuvrability of scaffold from the surface of electrodes precluded measurements on cardiac cells. In addition, COMSOL Multiphysics software was used to investigate the mechanical and electrical forces theoretically acting on the cells. Thirdly, in this thesis the cardiac electrophysiology was investigated using immunostaining techniques to visualize the growth of sarcomeres and gap junctions that promote cell-cell interaction and excitation-contraction of heart muscles. The physiological response of beating of co-cultured cardiomyocytes and cardiac fibroblasts was observed in a synchronous and simultaneous manner closely mimicking the native cardiac impulses. Further investigations were carried out by mechanically stimulating the cells in the three-dimensional hydrogel using standing surface acoustic waves and comparing with traditional two-dimensional flat surface coated with fibronectin. The electrophysiological responses of the cells under the effect of the mechanical stimulations yielded a higher magnitude of contractility, action potential and calcium transient.

Peripheral chemoreceptors and cardiorespiratory coupling: a link to sympatho-excitation

Chronic intermittent hypoxia (CIH) has been identified as a relevant risk factor for the development of enhanced sympathetic outflow and arterial hypertension. Several studies have highlighted the importance of peripheral chemoreceptors for the cardiovascular changes elicited by CIH. However, the effects of CIH on the central mechanisms regulating sympathetic outflow are not fully elucidated. Our research group has explored the hypothesis that the enhanced sympathetic drive following CIH exposure is, at least in part, dependent on alterations in the respiratory network and its interaction with the sympathetic nervous system. In this report, I discuss the changes in the discharge profile of baseline sympathetic activity in rats exposed to CIH, their association with the generation of active expiration and the interactions between expiratory and sympathetic neurones after CIH conditioning. Together, these findings are consistent with the theory that mechanisms of central respiratory–sympathetic coupling are a novel factor in the development of neurogenic hypertension.

Indirect coupling of phosphate release to de novo tension generation during muscle contraction.

A key question in muscle contraction is how tension generation is coupled to the chemistry of the actomyosin ATPase. Biochemical and mechanochemical experiments link tension generation to a change in structure associated with phosphate release. Length-jump and temperature-jump experiments, on the other hand, implicate phase 2slow, a significantly faster, markedly strain-sensitive kinetic process in tension generation. We use a laser temperature jump to probe the kinetics and mechanism of tension generation in skinned rabbit psoas fibers--an appropriate method since both phosphate release and phase 2slow are readily perturbed by temperature. Kinetics characteristic of the structural change associated with phosphate release are observed only when phosphate is added to fibers. When present, it causes a reduction in fiber tension; otherwise, no force is generated when it is perturbed. We therefore exclude this step from tension generation. The kinetics of de novo tension generation by the temperature-jump equivalent of phase 2slow appear unaffected by phosphate binding. We therefore propose that phosphate release is indirectly coupled to de novo tension generation via a steady-state flux through an irreversible step. We conclude that tension generation occurs in the absence of chemical change as the result of an entropy-driven transition between strongly bound crossbridges in the actomyosin-ADP state. The mechanism resembles the operation of a clock, with phosphate release providing the energy to tension the spring, and the irreversible step functions as the escapement mechanism, which is followed in turn by tension generation as the movement of the hands.

Thermodynamic origin of cooperativity in actomyosin interactions: The coupling of short-range interactions with actin bending stiffness in an Ising-like model

We present Monte Carlo simulations for a molecular motor system found in virtually all eukaryotic cells, the acto-myosin motor system, composed of a group of organic macromolecules. Cell motors were mapped to an Ising-like model, where the interaction field is transmitted through a tropomyosin polymer chain. The presence of Ca(2+) induces tropomyosin to block or unblock binding sites of the myosin motor leading to its activation or deactivation. We used the Metropolis algorithm to find the transient and the equilibrium states of the acto-myosin system composed of solvent, actin, tropomyosin, troponin, Ca(2+), and myosin-S1 at a given temperature, including the spatial configuration of tropomyosin on the actin filament surface. Our model describes the short- and long-range cooperativity during actin-myosin binding which emerges from the bending stiffness of the tropomyosin complex. We found all transition rates between the states only using the interaction energy of the constituents. The agreement between our model and experimental data also supports the recent theory of flexible tropomyosin.

Breakup coupling effects on near-barrier quasi-elastic scattering of (6,7)Li on (144)Sm

Excitation functions of quasi-elastic scattering at backward angles have been measured for the (6,7)Li + (144)Sm systems at near-barrier energies, and fusion barrier distributions have been extracted from the first derivatives of the experimental cross sections with respect to the bombarding energies. The data have been analyzed in the framework of continuum discretized coupled-channel calculations, and the results have been obtained in terms of the influence exerted by the inclusion of different reaction channels, with emphasis on the role played by the projectile breakup.

Acoustic Simulation of a Special Switched Reluctance Drive by Means of Field-Circuit Coupling and Multiphysics Simulation

The approach presented in this paper consists of an energy-based field-circuit coupling in combination with multi-physics simulation of the acoustic radiation of electrical machines. The proposed method is applied to a special switched reluctance motor with asymmetric pole geometry to improve the start-up torque. The pole shape has been optimized, subject to low torque ripple, in a previous study. The proposed approach here is used to analyze the impact of the optimization on the overall acoustic behavior. The field-circuit coupling is based on a temporary lumped-parameter model of the magnetic part incorporated into a circuit simulation based on the modified nodal analysis. The harmonic force excitation is calculated by means of stress tensor computation, and it is transformed to a mechanical mesh by mapping techniques. The structural dynamic problem is solved in the frequency domain using a finite-element modal analysis and superposition. The radiation characteristic is obtained from boundary element acoustic simulation. Simulation results of both rotor types are compared, and measurements of the drive are presented.

Both beta(2)- and beta(1)-adrenergic receptors mediate hastened relaxation and phosphorylation of phospholamban and troponin I in ventricular myocardium of fallot infants, consistent with selective coupling of beta(2)-adrenergic receptors to G(s)-protein

Background-In adult human heart, both beta(1)- and beta(2)-adrenergic receptors mediate hastening of relaxation; however, it is unknown whether this also occurs in infant heart. We compared the effects of stimulation of beta(1)- and beta(2)-adrenergic receptors on relaxation and phosphorylation of phospholamban and troponin I in ventricle obtained from infants with tetralogy of Fallot. Methods and Results-Myocardium dissected from the right ventricular outflow tract of 27 infants (age range 2-1/2 to 35 months) with tetralogy of Fallot was set up to contract 60 times per minute. Selective stimulation of beta(1)-adrenergic receptors with (-)-norepinephrine (NE) and beta(2)-adrenergic receptors with (-)-epinephrine (EPI) evoked phosphorylation of phospholamban (at serine-16 and threonine-17) and troponin I and caused concentration-dependent increases in contractile force (-log EC50 [mol/L] NE 5.5+/-0.1, n=12; -EPI 5.6+/-0.1, n=13 patients), hastening of the time to reach peak force (-log EC50 [mol/L] NE 5.8+/--0.2; EPI 5.8+/-0.2) and 50% relaxation (-log EC50 [mol/L] NE 5.7+/-0.2: EPI 5.8+/-0.1), Ventricular membranes from Fallot infants, labeled with (-)-[I-125]-cyanopindolol, revealed a greater percentage of beta(1)- (71%) than beta(2)-adrenergic receptors (29%). Binding of (-)-epinephrine to beta(2)-receptors underwent greater GTP shifts than binding of (-)-norepinephrine to beta(1)-receptors. Conclusions-Despite their low density, beta(2)-adrenergic receptors are nearly as effective as beta(1)-adrenergic receptors of infant Fallot ventricle in enhancing contraction, relaxation, and phosphorylation of phospholamban and troponin I, consistent with selective coupling to G(s)-protein.

Ground-state order and spin-lattice coupling in tetrahedral spin systems Cu2Te2O5X2 (X = Br or Cl)

High-resolution ac susceptibility and thermal conductivity measurement on Cu2Te2O5X2 (X=Br,Cl) single crystals are reported. For Br-sample, sample dependence prevents one from distinguishing between possibilities of magnetically ordered and spin-singlet ground states. In Cl-sample a three-dimensional transition at 18.5 K is accompanied by almost isotropic behavior of susceptibility and almost switching behavior of thermal conductivity. Thermal conductivity studies suggest the presence of a tremendous spin-lattice coupling characterizing Cl- but not Br-sample. Below the transition Cl-sample is in a complex magnetic state involving AF order but also the elements consistent with the presence of a gap in the excitation spectrum.

Experimental Vibration Characteristics of a Beam with Nonlinear Support Using Receptance Coupling Analysis

This paper applies the Multi-Harmonic Nonlinear Receptance Coupling Approach (MUHANORCA) (Ferreira 1998) to evaluate the frequency response characteristics of a beam which is clamped at one end and supported at the other end by a nonlinear cubic stiffness joint. In order to apply the substructure coupling technique, the problem was characterised by coupling a clamped linear beam with a nonlinear cubic stiffness joint. The experimental results were obtained by a sinusoidal excitation with a special force control algorithm where the level of the fundamental force is kept constant and the level of the harmonics is kept zero for all the frequencies measured.

Excitation of low-profile equilateral-triangular dielectric resonator antenna using a conducting conformal strip

A new configuration that employs a conducting conformal strip to excite the low-profile equilaterial-triangular dielectric resonator antenna (DRA) of very high permittivity is proposed. As compared with the previous aperture-coupling configuration, the new configuration has a wider impedance bandwidth (- 5.5%) and a higher front-to-back radiation ratio. The return loss, radiation patterns, and antenna gain are measured and discussed

New calculations of P(b) curves for 1s_\sigma excitation in low-Z (Z\le10) ion-atom scattering

Ab initio fully relativistic SCF molecular calculations of energy eigenvalues as well as coupling-matrix elements are used to calculate the 1s_\sigma excitation differential cross section for Ne-Ne and Ne-O in ion-atom collisions. A relativistic perturbation treatment which allows a direct comparison with analogous non-relativistic calculations is also performed.

Equilibrium K-shell excitation of highly ionized neon

Augerelectron emission from foil-excited Ne-ions (6 to 10 MeV beam energy) has been measured. The beam-foil time-of-flight technique has been applied to study electronic transitions of metastable states (delayed spectra) and to determine their lifetimes. To achieve a line identification for the complex structure observed in the prompt spectrum, the spectrum is separated into its isoelectronic parts by an Augerelectron-ion coincidence correlating the emitted electrons and the emitting projectiles of well defined final charge states q_f. Well resolved spectra were obtained and the lines could be identified using intermediate coupling Dirac-Fock multiconfiguration calculations. From the total KLL-Augerelectron transition probabilities observed in the electronion coincidence experiment for Ne (10 MeV) the amount of projectiles with one K-hole just behind a C-target can be estimated. For foil-excited Ne-projectiles in contrast to single collision results the comparison of transition intensities for individual lines with calculated transition probabilities yields a statistical population of Li- and Be-like configurations.

Thermodynamics/dynamics coupling in weakly compressible turbulent stratified fluids

In traditional and geophysical fluid dynamics, it is common to describe stratified turbulent fluid flows with low Mach number and small relative density variations by means of the incompressible Boussinesq approximation. Although such an approximation is often interpreted as decoupling the thermodynamics from the dynamics, this paper reviews recent results and derive new ones that show that the reality is actually more subtle and complex when diabatic effects and a nonlinear equation of state are retained. Such an analysis reveals indeed: (1) that the compressible work of expansion/contraction remains of comparable importance as the mechanical energy conversions in contrast to what is usually assumed; (2) in a Boussinesq fluid, compressible effects occur in the guise of changes in gravitational potential energy due to density changes. This makes it possible to construct a fully consistent description of the thermodynamics of incompressible fluids for an arbitrary nonlinear equation of state; (3) rigorous methods based on using the available potential energy and potential enthalpy budgets can be used to quantify the work of expansion/contraction B in steady and transient flows, which reveals that B is predominantly controlled by molecular diffusive effects, and act as a significant sink of kinetic energy.

Excitation and decay of solar-wind driven flows in the magnetosphere-ionosphere system

Basic concepts of the form of high-latitude ionospheric flows and their excitation and decay are discussed in the light of recent high time-resolution measurements made by ground-based radars. It is first pointed out that it is in principle impossible to adequately parameterize these flows by any single quantity derived from concurrent interplanetary conditions. Rather, even at its simplest, the flow must be considered to consist of two basic time-dependent components. The first is the flow driven by magnetopause coupling processes alone, principally by dayside reconnection. These flows may indeed be reasonably parameterized in terms of concurrent near-Earth interplanetary conditions, principally by the interplanetary magnetic field (IMF) vector. The second is the flow driven by tail reconnection alone. As a first approximation these flows may also be parameterized in terms of interplanetary conditions, principally the north-south component of the IMF, but with a delay in the flow response of around 30-60 min relative to the IMF. A delay in the tail response of this order must be present due to the finite speed of information propagation in the system, and we show how "growth" and "decay" of the field and flow configuration then follow as natural consequences. To discuss the excitation and decay of the two reconnection-driven components of the flow we introduce that concept of a flow-free equilibrium configuration for a magnetosphere which contains a given (arbitrary) amount of open flux. Reconnection events act either to create or destroy open flux, thus causing departures of the system from the equilibrium configuration. Flow is then excited which moves the system back towards equilibrium with the changed amount of open flux. We estimate that the overall time scale associated with the excitation and decay of the flow is about 15 min. The response of the system to both impulsive (flux transfer event) and continuous reconnection is discussed in these terms.

The excitation of ionospheric convection

This review presents recent observations of high-latitude ionospheric plasma convection, obtained using the EISCAT radar in the 'Polar' experiment mode. The paper is divided into two main parts. Firstly, the delay in the response of dayside high-latitude flows to changes in the interplanetary magnetic field is discussed. The results show the importance for the excitation of dayside convection of the transfer of magnetic flux from the dayside into the tail lobe. Consequently, ionospheric convection should be thought of as the sum of two intrinsically time-dependent flow patterns. The first of these patterns is directly driven by solar wind-magnetosphere coupling, dominates ionospheric flows on the dayside, is associated with an expanding polar cap area and is the F-region flow equivalent of the DP-2 E-region current system. The second of the two patterns is driven by the release of energy stored in the geomagnetic tail, dominates ionospheric flows on the nightside, is associated with a contracting polar cap and is equivalent to the DP-1, or substorm, current system. In the second half of the paper, various transient flow bursts observed in the vicinity of the dayside cusp are studied. These radar data, combined with simultaneous optical observations of transient dayside aurorae, strongly suggest that momentum is transferred across the magnetopause and into the ionosphere in a series of bursts, each associated with voltages of 30-80 kV. Similarities between these bursts and flux transfer events observed at the magnetopause are discussed.