The fluorolysis assay, a highly sensitive method for measuring the cytolytic activity of T cells at very low numbers

We have developed a highly sensitive cytolysis test, the fluorolysis assay, as a simple nonradioactive and inexpensive alternative to the standard Cr-51-release assay. P815 cells were stably transfected with a plasmid expressing the enhanced green fluorescent protein (EGFP) gene. These target cells were coated with or without cognate peptide or anti-CD3 Ab and then incubated with CD8(+) T cells to allow antigen-specific or nonspecific lysis. The degree of target cell lysis was measured using flow cytometry to count the percentage of viable propidium iodide(-) EGFP(+) cells, whose numbers were standardized to a reference number of fluorochrome-linked beads. By using small numbers of target cells (200-800 per reaction) and extended incubation times (up to 2 days), the antigen-specific cytolytic activity of one to two activated CD8(+) T cells of a CTL line could be detected. The redirected fluorolysis assay also measured the activity of very few ( greater than or equal to6) primary CD8(+) T cells following polyclonal activation. Importantly, antigen-specific lysis by small numbers ( greater than or equal to 25) of primary CD8(+) T cells could be directly measured ex vivo. This exquisite sensitivity of the fluorolysis assay, which was at least 8-33-folds higher than an optimized 51 Cr-release assay, allows in vitro and ex vivo studies of immune responses that would otherwise not be possible due to low CTL numbers or frequencies. (C) 2002 Elsevier Science B.V. All rights reserved.

On the fast approximation of Green's functions in MPIE formulations for planar layered media

The numerical implementation of the complex image approach for the Green's function of a mixed-potential integralequation formulation is examined and is found to be limited to low values of k(0) rho (in this context k(0) rho = 2 pirho/ lambda(0), where rho is the distance between the source and the field points of the Green's function and lambda(0) is the free space wavelength). This is a clear limitation for problems of large dimension or high frequency where this limit is easily exceeded. This paper examines the various strategies and proposes a hybrid method whereby most of the above problems can be avoided. An efficient integral method that is valid for large k(0) rho is combined with the complex image method in order to take advantage of the relative merits of both schemes. It is found that a wide overlapping region exists between the two techniques allowing a very efficient and consistent approach for accurately calculating the Green's functions. In this paper, the method developed for the computation of the Green's function is used for planar structures containing both lossless and lossy media.

Marx as a Critical Discourse Analyst: The genesis of a critical method and its relevance to the critique of global capital

In this paper we identify elements in Marx´s economic and political writings that are relevant to contemporary critical discourse analysis (CDA). We argue that Marx can be seen to be e n gaging in a form of discourse analysis. We identify the elements in Marx´s historical materialist method that support such a perspective, and exemplify these in a longitudinal comparison of Marx´s texts.

Polarization squeezing and continuous-variable polarization entanglement

A concept of polarization entanglement for continuous variables is introduced. For this purpose the Stokes-parameter operators and the associated Poincare sphere, which describe the quantum-optical polarization properties of light, are defined and their basic properties are reviewed. The general features of the Stokes operators are illustrated by evaluation of their means and variances for a range of simple polarization states. Some of the examples show polarization squeezing, in which the variances of one or more Stokes parameters are smaller than the coherent-state value. The main object of the paper is the application of these concepts to bright squeezed light. It is shown that a light beam formed by interference of two orthogonally polarized quadrature-squeezed beams exhibits squeezing in some of the Stokes parameters. Passage of such a primary polarization-squeezed beam through suitable optical components generates a pair of polarization-entangled light beams with the nature of a two-mode squeezed state. Implementation of these schemes using the double-fiber Sagnac interferometer provides an efficient method for the generation of bright nonclassical polarization states. The important advantage of these nonclassical polarization states for quantum communication is the possibility of experimentally determining all of the relevant conjugate variables of both squeezed and entangled fields using only linear optical elements followed by direct detection.

R-matrices and the tensor product graph method

A systematic method for constructing trigonometric R-matrices corresponding to the (multiplicity-free) tensor product of any two affinizable representations of a quantum algebra or superalgebra has been developed by the Brisbane group and its collaborators. This method has been referred to as the Tensor Product Graph Method. Here we describe applications of this method to untwisted and twisted quantum affine superalgebras.

Control of chaotic instabilities in a spinning spacecraft with dissipation using Lyapunov's method

Control of chaotic instability in a simplified model of a spinning spacecraft with dissipation is achieved using an algorithm derived using Lyapunov's second method. The control method is implemented on a realistic spacecraft parameter configuration which has been found to exhibit chaotic instability for a range of forcing amplitudes and frequencies when a sinusoidally varying torque is applied to the spacecraft. Such a torque, may arise in practice from an unbalanced rotor or from vibrations in appendages. Numerical simulations are performed and the results are studied by means of time history, phase space, Poincare map, Lyapunov characteristic exponents and bifurcation diagrams. (C) 2002 Elsevier Science Ltd. All rights reserved.

A monolithic smoothing-gap algorithm for contact-impact based on the signed distance function

A new algorithm has been developed for smoothing the surfaces in finite element formulations of contact-impact. A key feature of this method is that the smoothing is done implicitly by constructing smooth signed distance functions for the bodies. These functions are then employed for the computation of the gap and other variables needed for implementation of contact-impact. The smoothed signed distance functions are constructed by a moving least-squares approximation with a polynomial basis. Results show that when nodes are placed on a surface, the surface can be reproduced with an error of about one per cent or less with either a quadratic or a linear basis. With a quadratic basis, the method exactly reproduces a circle or a sphere even for coarse meshes. Results are presented for contact problems involving the contact of circular bodies. Copyright (C) 2002 John Wiley Sons, Ltd.

An assessment of maturity from anthropometric measurements

Purpose: The range of variability between individuals of the same chronological age (CA) in somatic and biological maturity is large and especially accentuated around the adolescent growth spurt. Maturity assessment is an important consideration when dealing with adolescents, from both a research perspective and youth sports stratification. A noninvasive, practical method predicting years from peak height velocity (a maturity offset value) by using anthropometric variables is developed in one sample and cross-validated in two different samples. Methods: Gender specific multiple regression equations were calculated on a sample of 152 Canadian children aged 8-16 yr (79 boys; 73 girls) who were followed through adolescence from 1991 to 1997, The equations included three somatic dimensions (height, sitting height, and leg length), CA, and their interactions. The equations were cross-validated on a Combined sample of Canadian (71 boys, 40 girls measured from 1964 through 1973) and Flemish children (50 boys, 48 girls measured from 1985 through 1999). Results: The coefficient of determination (R2) for the boys' model was 0.92 and for the girls' model 0.91 the SEEs were 0.49 and 0.50, respectively, Mean difference between actual and predicted maturity offset for the verification samples was 0.24 (SD 0.65) yr in boys and 0,001 (SD 0.68) yr in girls. Conclusion: Although the cross-validation meets statistical standards or acceptance, caution 1, warranted with regard to implementation. It is recommended that maturity offset be considered as a categorical rather than a continuous assessment. Nevertheless, the equations presented are a reliable, noninvasive and a practical solution for the measure of biological maturity for matching adolescent athletes.

A variable stepsize implementation for stochastic differential equations

Stochastic differential equations (SDEs) arise from physical systems where the parameters describing the system can only be estimated or are subject to noise. Much work has been done recently on developing higher order Runge-Kutta methods for solving SDEs numerically. Fixed stepsize implementations of numerical methods have limitations when, for example, the SDE being solved is stiff as this forces the stepsize to be very small. This paper presents a completely general variable stepsize implementation of an embedded Runge Kutta pair for solving SDEs numerically; in this implementation, there is no restriction on the value used for the stepsize, and it is demonstrated that the integration remains on the correct Brownian path.

A four-stage index 2 Diagonally Implicit Runge-Kutta method

High index Differential Algebraic Equations (DAEs) force standard numerical methods to lower order. Implicit Runge-Kutta methods such as RADAU5 handle high index problems but their fully implicit structure creates significant overhead costs for large problems. Singly Diagonally Implicit Runge-Kutta (SDIRK) methods offer lower costs for integration. This paper derives a four-stage, index 2 Explicit Singly Diagonally Implicit Runge-Kutta (ESDIRK) method. By introducing an explicit first stage, the method achieves second order stage calculations. After deriving and solving appropriate order conditions., numerical examples are used to test the proposed method using fixed and variable step size implementations. (C) 2001 IMACS. Published by Elsevier Science B.V. All rights reserved.

Application of a simplified method based on regular regime approach to determine the effective moisture diffusivity of mixture of low molecular weight sugars and maltodextrin during desorption

The binary diffusivities of water in low molecular weight sugars; fructose, sucrose and a high molecular weight carbohydrate; maltodextrin (DE 11) and the effective diffusivities of water in mixtures of these sugars (sucrose, glucose, fructose) and maltodextrin (DE 11) were determined using a simplified procedure based on the Regular Regime Approach. The effective diffusivity of these mixtures exhibited both the concentration and molecular weight dependence. Surface stickiness was observed in all samples during desorption, with fructose exhibiting the highest and maltodextrin the lowest. (C) 2002 Elsevier Science Ltd. All rights reserved.

Comparison of an ELISA and an LC/MS/MS method for measuring tacrolimus concentrations and making dosage decisions in transplant recipients

This study compared an enzyme-linked immunosorbent assay (ELISA) to a liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique for measurement of tacrolimus concentrations in adult kidney and liver transplant recipients, and investigated how assay choice influenced pharmacokinetic parameter estimates and drug dosage decisions. Tacrolimus concentrations measured by both ELISA and LC/MS/MS from 29 kidney (n = 98 samples) and 27 liver (n = 97 samples) transplant recipients were used to evaluate the performance of these methods in the clinical setting. Tacrolimus concentrations measured by the two techniques were compared via regression analysis. Population pharmacokinetic models were developed independently using ELISA and LC/MS/MS data from 76 kidney recipients. Derived kinetic parameters were used to formulate typical dosing regimens for concentration targeting. Dosage recommendations for the two assays were compared. The relation between LC/MS/MS and ELISA measurements was best described by the regression equation ELISA = 1.02 . (LC/MS/MS) + 0.14 in kidney recipients, and ELISA = 1.12 . (LC/MS/MS) - 0.87 in liver recipients. ELISA displayed less accuracy than LC/MS/MS at lower tacrolimus concentrations. Population pharmacokinetic models based on ELISA and LC/MS/MS data were similar with residual random errors of 4.1 ng/mL and 3.7 ng/mL, respectively. Assay choice gave rise to dosage prediction differences ranging from 0% to 30%. ELISA measurements of tacrolimus are not automatically interchangeable with LC/MS/MS values. Assay differences were greatest in adult liver recipients, probably reflecting periods of liver dysfunction and impaired biliary secretion of metabolites. While the majority of data collected in this study suggested assay differences in adult kidney recipients were minimal, findings of ELISA dosage underpredictions of up to 25% in the long term must be investigated further.