Analysis of a conformation-specific epitope of the alpha subunit of human chorionic gonadotropin: Study using monoclonal antibody probes

Monoclonal antibodies (MAbs) have been used extensively for identification of sequence-specific epitopes using either the ELISA or/and IRMA methods, However, attempts to use MAbs for identification of conformation-specific epitopes have been very few as they are considered very labile. We have investigated the stability of conformation-specific epitopes of human chorionic gonadotropin (hCG) using a quantitative solid-phase radioimmnunoassay (SPRIA) technique. Several epitopes are stable to mild modification (chemical and proteolytic) conditions, and epitopes show differential stability for these modifications. Based on these observations, a monoclonal antibody (MAb 16) for an a-subunit-specific epitope of hCG has been used to monitor changes at the epitopic site (identified as epitope 16) on modification of hCG, using SPRIA with immobilized MAb 16. Modifications of amino groups, hydroxyl group of tyrosine as well as carboxyl group of Asp/Glu all bring about sufficient changes in the epitope integrity. Peptide bond hydrolysis at lysine residues damages the epitope, but not at arginine residues, Hydrolysis at tyrosine does not affect the epitope, though modification of the side-chain of tyrosine inactivates the epitope. Destruction of the epitope occurs on reduction of the disulphide bonds. Partial retention of the epitope activity is seen on modification of carboxyl or the epsilon-amino groups of lysine. Based on these results four to six amino acids have been identified to be at the epitopic site, and the data suggest that two peptide segments are brought together by the disulphide bond Cys10-Cys60 to form the epitope.

Flap-lag-torsion stability in hover and forward flight with three-dimensional wake

The contributions of full-wake dynamics in trim analysis are demonstrated for finding the control inputs and periodic responses simultaneously, as well as in Floquet eigenanalysis for finding the damping levels. The equations of flap bending, lag bending, and torsion are coupled with a three-dimensional, finite state wake, and low-frequency (<1/rev) to high frequency (>1/rev) multiblade modes are considered. Full blade-wake dynamics is used in trim analysis and Floquet eigenanalysis. A uniform cantilever blade in trimmed flight is investigated over a range of thrust levels, advance ratios, number of blades, and blade torsional frequencies. The investigation includes the convergence characteristics of control inputs, periodic responses, and damping levels with respect to the number of spatial azimuthal harmonics and radial shape functions in the wake representation. It also includes correlation with the measured lag damping of a three-bladed untrimmed rotor. The parametric study shows the dominant influence of wake dynamics on control inputs, periodic responses, and damping levels, and wake theory generally improves the correlation.

Stability of laminated composite plates with cut-outs

Critical buckling loads of laminated fibre-reinforced plastic square panels have been obtained using the finite element method. Various boundary conditions, lay-up details, fibre orientations, cut-out sizes are considered. A 36 degrees of freedom triangular element, based on the classical lamination theory (CLT) has been used for the analysis. The performance of this element is validated by comparing results with some of those available in literature. New results have been given for several cases of boundary conditions for [0°/ ± 45°/90°]s laminates. The effect of fibre-orientation in the ply on the buckling loads has been investigated by considering [±?]6s laminates.

Engineered dimer interface mutants of triosephosphate isomerase: the role of inter-subunit interactions in enzyme function and stability

The role of inter-subunit interactions in maintaining optimal catalytic activity in triosephosphate isomerase (TIM) has been probed, using the Plasmodium falciparum enzyme as a model. Examination of subunit interface contacts in the crystal structures suggests that residue 75 (Thr, conserved) and residue 13 (Cys, variable) make the largest number of inter-subunit contacts. The mutants Cys13Asp (C13D) and Cys13Glu (C13E) have been constructed and display significant reduction in catalytic activity when compared with wild-type (WT) enzyme (similar to 7.4-fold decrease in k(cat) for the C13D and similar to 3.3-fold for the C13E mutants). Analytical gel filtration demonstrates that the C13D mutant dissociates at concentrations < 1.25 mu M, whereas the WT and the C13E enzymes retain the dimeric structure. The order of stability of the mutants in the presence of chemical denaturants, like urea and guanidium chloride, is WT > Cys13Glu > Cys13Asp. Irreversible thermal precipitation temperatures follow the same order as well. Modeling studies establish that the Cys13Asp mutation is likely to cause a significantly greater structural perturbation than Cys13Glu. Analysis of sequence and structural data for TIMs from diverse sources suggests that residues 13 and 82 form a pair of proximal sites, in which a limited number of residue pairs may be accommodated.

Microstructural stability and nanoindentation of an electrodeposited nanocrystalline Ni-1.5wt.%P alloy

Microstructural stability of nanocrystalline Ni-1.5wt.%P alloy with an initial grain size of 3 nm processed by pulsed electrodeposition was studied using differential scanning calorimetry (DSC) and annealing. Microstructural characterization suggests that the observed exothermic peak during heating in DSC is related to both concurrent grain growth and Ni3P formation. Nanoindentation on samples with grain sizes from 3 to 50 nm revealed a breakdown in Hall-Petch strengthening in nano Ni-P alloy at grain sizes <= 10 nm, consistent with some previous observations. It is concluded that there is a grain boundary weakening regime for grain sizes < 10 nm, based on analysis which show that the data cannot be rationalized in terms of microstrain relaxation, variation in elastic modulus, texture evolution and duplex structure formation.

Stability of spatially developing boundary layers in pressure gradients

A new formulation of the stability of boundary-layer flows in pressure gradients is presented, taking into account the spatial development of the flow and utilizing a special coordinate transformation. The formulation assumes that disturbance wavelength and eigenfunction vary downstream no more rapidly than the boundary-layer thickness, and includes all terms nominally of order R(-1) in the boundary-layer Reynolds number R. In Blasius flow, the present approach is consistent with that of Bertolotti et al. (1992) to O(R(-1)) but simpler (i.e. has fewer terms), and may best be seen as providing a parametric differential equation which can be solved without having to march in space. The computed neutral boundaries depend strongly on distance from the surface, but the one corresponding to the inner maximum of the streamwise velocity perturbation happens to be close to the parallel flow (Orr-Sommerfeld) boundary. For this quantity, solutions for the Falkner-Skan flows show the effects of spatial growth to be striking only in the presence of strong adverse pressure gradients. As a rational analysis to O(R(-1)) demands inclusion of higher-order corrections on the mean flow, an illustrative calculation of one such correction, due to the displacement effect of the boundary layer, is made, and shown to have a significant destabilizing influence on the stability boundary in strong adverse pressure gradients. The effect of non-parallelism on the growth of relatively high frequencies can be significant at low Reynolds numbers, but is marginal in other cases. As an extension of the present approach, a method of dealing with non-similar flows is also presented and illustrated. However, inherent in the transformation underlying the present approach is a lower-order non-parallel theory, which is obtained by dropping all terms of nominal order R(-1) except those required for obtaining the lowest-order solution in the critical and wall layers. It is shown that a reduced Orr-Sommerfeld equation (in transformed coordinates) already contains the major effects of non-parallelism.

Stability of the viscous flow of a fluid through a flexible tube

The stability of the Hagen-Poiseuille flow of a Newtonian fluid in a tube of radius R surrounded by an incompressible viscoelastic medium of radius R < r < HR is analysed in the high Reynolds number regime. The dimensionless numbers that affect the fluid flow are the Reynolds number Re = (rho VR/eta), the ratio of the viscosities of the wall and fluid eta(r) = (eta(s)/eta), the ratio of radii H and the dimensionless velocity Gamma = (rho V-2/G)(1/2). Here rho is the density of the fluid, G is the coefficient of elasticity of the wall and V is the maximum fluid velocity at the centre of the tube. In the high Reynolds number regime, an asymptotic expansion in the small parameter epsilon = (1/Re) is employed. In the leading approximation, the viscous effects are neglected and there is a balance between the inertial stresses in the fluid and the elastic stresses in the medium. There are multiple solutions for the leading-order growth rate s((0)), all of which are imaginary, indicating that the fluctuations are neutrally stable, since there is no viscous dissipation of energy or transfer of energy from the mean flow to the fluctuations due to the Reynolds stress. There is an O(epsilon(1/2)) correction to the growth rate, s((1)), due to the presence of a wall layer of thickness epsilon(1/2)R where the viscous stresses are O(epsilon(1/2)) smaller than the inertial stresses. An energy balance analysis indicates that the transfer of energy from the mean flow to the fluctuations due to the Reynolds stress in the wall layer is exactly cancelled by an opposite transfer of equal magnitude due to the deformation work done at the interface, and there is no net transfer from the mean flow to the fluctuations. Consequently, the fluctuations are stabilized by the viscous dissipation in the wall layer, and the real part of s(1) is negative. However, there are certain values of Gamma and wavenumber k where s((1)) = 0. At these points, the wall layer amplitude becomes zero because the tangential velocity boundary condition is identically satisfied by the inviscid flow solution. The real part of the O(epsilon) correction to the growth rate s((2)) turns out to be negative at these points, indicating a small stabilizing effect due to the dissipation in the bulk of the fluid and the wall material. It is found that the minimum value of s((2)) increases proportional to (H-1)(-2) for (H-1) much less than 1 (thickness of wall much less than the tube radius), and decreases proportional to H-4 for H much greater than 1. The damping rate for the inviscid modes is smaller than that for the viscous wall and centre modes in a rigid tube, which have been determined previously using a singular perturbation analysis. Therefore, these are the most unstable modes in the flow through a flexible tube

Stochastic Aeroelastic Analysis of Composite Helicopter Rotor

The effect of uncertainty in composite material properties on the aeroelastic response, vibratory loads, and stability of a hingeless helicopter rotor is investigated. The uncertainty impact on rotating natural frequencies of the blade is studied with Monte Carlo simulations and first-order reliability methods. The stochastic aeroelastic analyses in hover and forward flight are carried out with Monte Carlo simulations. The flap, lag, and torsion responses show considerable scatter from their baseline values, and the uncertainty impact varies with the azimuth angle. Furthermore, the blade response shows finite probability of resonance-type conditions caused by modal frequencies approaching multiples of the rotor speed. The 4/rev vibratory forces show large deviations from their baseline values. The lag mode damping shows considerable scatter due to uncertain material properties with an almost 40% probability of instability in hover.

Microstructural stability and superplasticity in an electrodeposited nanocrystalline Ni-P alloy

Stabilization of nanocrystalline grain sizes by second phase particles can facilitate superplasticity at high strain rates and/or low temperatures. A metastable single phase nano-Ni-P alloy prepared by electrodeposition, with a grain size of similar to 6 nm, transforms to a nanoduplex structure at T> 673 K, with similar to 4 vol.% Ni3P particles at triple junctions and within Ni grains. The nanoduplex microstructure is reasonably stable up to 777 K, and the growth of Ni grains occurs in a coupled manner with the growth of Ni3P particles such that the ratio of the two mean sizes (Z) is essentially constant. High temperature tests for a grain size of 290 nm reveal superplastic behavior with an optimum elongation to failure of 810% at a strain rate of 7 x 10(-4) s(-1) and a relatively low temperature of 777 K. Superplastic deformation enhances both grain growth and the ratio Z, implying that grain boundary sliding (GBS) significantly influences the microstructural dynamics. Analysis of the deformation processes suggests that superplasticity is associated with GBS controlled by the overcoming of intragranular particles by dislocations, so that deformation is independent of the grain size. The nano-Ni-P alloy exhibits lower ductility than nano-Ni due to concurrent cavitation caused by higher stresses. (C) 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Nonlinear analysis of adhesively bonded lap joints considering viscoplasticity in adhesives

This paper presents nonlinear finite element analysis of adhesively bonded joints considering the elastoviscoplastic constitutive model of the adhesive material and the finite rotation of the joint. Though the adherends have been assumed to be linearly elastic, the yielding of the adhesive is represented by a pressure sensitive modified von Mises yield function. The stress-strain relation of the adhesive is represented by the Ramberg-Osgood relation. Geometric nonlinearity due to finite rotation in the joint is accounted for using the Green-Lagrange strain tensor and the second Piola-Kirchhoff stress tensor in a total Lagrangian formulation. Critical time steps have been calculated based on the eigenvalues of the transition matrices of the viscoplastic model of the adhesive. Stability of the viscoplastic solution and time dependent behaviour of the joints are examined. A parametric study has been carried out with particular reference to peel and shear stress along the interface. Critical zones for failure of joints have been identified. The study is of significance in the design of lap joints as well as on the characterization of adhesive strength. (C) 1999 Elsevier Science Ltd. All rights reserved.

Stability of degenerate diffusions with state-dependent switching

This paper deals with the ergodic properties of hybrid systems modelled by diffusion processes with state-dependent switching. We investigate the sufficient conditions expressed in terms of the parameters of the underlying process which would ensure the existence of a unique invariant probability and stability in distribution of the flow. It turns out that the conditions would depend on certain averaging mechanisms over the states of the discrete component of the hybrid system. (C) 1999 Academic Press.

Weakly non-linear stability of a hydrodynamic accretion disc

When the cold accretion disc coupling between neutral gas and a magnetic field is so weak that the magnetorotational instability is less effective or even stops working, it is of prime interest to investigate the pure hydrodynamic origin of turbulence and transport phenomena. As the Reynolds number increases, the relative importance of the non-linear term in the hydrodynamic equation increases. In an accretion disc where the molecular viscosity is too small, the Reynolds number is large enough for the non-linear term to have new effects. We investigate the scenario of the `weakly non-linear' evolution of the amplitude of the linear mode when the flow is bounded by two parallel walls. The unperturbed flow is similar to the plane Couette flow, but with the Coriolis force included in the hydrodynamic equation. Although there is no exponentially growing eigenmode, because of the self-interaction, the least stable eigenmode will grow in an intermediate phase. Later, this will lead to higher-order non-linearity and plausible turbulence. Although the non-linear term in the hydrodynamic equation is energy-conserving, within the weakly non-linear analysis it is possible to define a lower bound of the energy (alpha A(c)(2), where A(c) is the threshold amplitude) needed for the flow to transform to the turbulent phase. Such an unstable phase is possible only if the Reynolds number >= 10(3-4). The numerical difficulties in obtaining such a large Reynolds number might be the reason for the negative result of numerical simulations on a pure hydrodynamic Keplerian accretion disc.

Optimum design of a helicopter rotor for low vibration using aeroelastic analysis and response surface methods

An aeroelastic analysis based on finite elements in space and time is used to model the helicopter rotor in forward flight. The rotor blade is represented as an elastic cantilever beam undergoing flap and lag bending, elastic torsion and axial deformations. The objective of the improved design is to reduce vibratory loads at the rotor hub that are the main source of helicopter vibration. Constraints are imposed on aeroelastic stability, and move limits are imposed on the blade elastic stiffness design variables. Using the aeroelastic analysis, response surface approximations are constructed for the objective function (vibratory hub loads). It is found that second order polynomial response surfaces constructed using the central composite design of the theory of design of experiments adequately represents the aeroelastic model in the vicinity of the baseline design. Optimization results show a reduction in the objective function of about 30 per cent. A key accomplishment of this paper is the decoupling of the analysis problem and the optimization problems using response surface methods, which should encourage the use of optimization methods by the helicopter industry. (C) 2002 Elsevier Science Ltd. All rights reserved.

Performance analysis of the random early detection algorithm

In this article we consider a finite queue with its arrivals controlled by the random early detection algorithm. This is one of the most prominent congestion avoidance schemes in the Internet routers. The aggregate arrival stream from the population of transmission control protocol sources is locally considered stationary renewal or Markov modulated Poisson process with general packet length distribution. We study the exact dynamics of this queue and provide the stability and the rates of convergence to the stationary distribution and obtain the packet loss probability and the waiting time distribution. Then we extend these results to a two traffic class case with each arrival stream renewal. However, computing the performance indices for this system becomes computationally prohibitive. Thus, in the latter half of the article, we approximate the dynamics of the average queue length process asymptotically via an ordinary differential equation. We estimate the error term via a diffusion approximation. We use these results to obtain approximate transient and stationary performance of the system. Finally, we provide some computational examples to show the accuracy of these approximations.

Reliability Based Design Optimization for Internal Stability of Reinforced Soil Structures Using Pseudo‐Static Method

Seismic design of reinforced soil structures involves many uncertainties that arise from the backfill soil properties and tensile strength of the reinforcement which is not addressed in current design guidelines. This paper highlights the significance of variability in the internal stability assessment of reinforced soil structures. Reliability analysis is applied to estimate probability of failure and pseudo‐static approach has been used for the calculation of the tensile strength and length of the reinforcement needed to maintain the internal stability against tension and pullout failures. Logarithmic spiral failure surface has been considered in conjunction with the limit equilibrium method. Two modes of failure namely, tension failure and pullout failure have been considered. The influence of variations of the backfill soil friction angle, the tensile strength of reinforcement, horizontal seismic acceleration on the reliability index against tension failure and pullout failure of reinforced earth structure have been discussed.