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.

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.

What is the value of firms' prior knowledge? Building organisational knowledge capabilities

A firm's prior knowledge facilitates the absorption of new knowledge, thereby renewing a firm's systematic search, transfer and absorption capabilities. The rapidly expanding field of biotechnology is characterised by the convergence of disparate sciences and technologies. This paper, the shift from proteinbased to DNA-based diagnostic technologies, quantifies the value of a firm's prior knowledge and its relation to future knowledge development. Four dimensions of diagnostic and four dimensions of knowledge in biotechnology firms are analysed. A simple scaled matrix method is developed to quantify the positive and negative heuristic values of prior scientific and technological knowledge that is useful for the acquisition and absorption of new knowledge.

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 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.

The absorption and excretion of fluoride and arsenic in humans

The absorption and excretion of fluoride and arsenic were measured in a group of healthy volunteers given drinking water with naturally high concentration of fluoride (F 2.3 mg/l), or high concentration of arsenic (As 0.15 mg/l), or high concentrations of both fluoride and arsenic (F 2.25 mg/l, As 0.23 mg/l and F 4.05 mg/l, As 0.58 mg/l), respectively. The results indicated that, for arsenic, the absorption rate, the proportion of urinary excretion and the biological-half-life did not show statistically significant differences between drinking water containing high arsenic alone and drinking water containing different levels of high arsenic and fluoride. Excretion and retention of arsenic were positively correlated to the total arsenic intake. Similar results were observed for fluoride. This suggests that there are different metabolic processes for arsenic and fluoride in respect to absorption and excretion; and no joint action can be attributed by these two elements. (C) 2002 Elsevier Science Ireland 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.

Linguistic evaluation method for the environmental assessment of infrastructure projects

A soft linguistic evaluation method is proposed for the environmental assessment of physical infrastructure projects based on fuzzy relations. Infrastructure projects are characterized in terms of linguistic expressions of 'performance' with respect to factors or impacts and the 'importance' of those factors/impacts. A simple example is developed to illustrate the method in the context of three road infrastructure projects assessed against five factors/impacts. In addition, a means to include hard or crisp factors is presented and illustrated with respect to a sixth factor.

Beneficial effects of air inclusions on the performance of ethylene vinyl acetate (EVA) mouthguard material

Objective: To investigate the impact characteristics of an ethylene vinyl acetate (EVA) mouthguard material with regulated air inclusions, which included various air cell volumes and wall thickness between air cells. In particular, the aim was to identify the magnitude and direction of forces within the impacts. Method: EVA mouthguard material, A mm thick and with and without air inclusions, was impacted with a constant force impact pendulum with an energy of 4.4 J and a velocity of 3 m/s. Transmitted forces through the EVA material were measured using an accelerometer, which also allowed the determination of force direction and magnitude within the impacts. Results: Statistically significant reductions in the transmitted forces were observed with all the air inclusion materials when compared with EVA without air inclusions. Maximum transmitted force through one air inclusion material was reduced by 32%. Force rebound was eliminated in one material, and reduced second force impulses were observed in all the air inclusion materials. Conclusion: The regulated air inclusions improved the impact characteristics of the EVA mouthguard material, the material most commonly used in mouthguards world wide.

Effect of ethylene vinyl acetate (EVA) closed cell foam on transmitted forces in mouthguard material

Objectives: To compare transmitted forces through ethylene vinyl acetate (EVA) mouthguard material and the same EVA material with gas inclusions in the form of a closed cell foam. Method: EVA mouthguard materials with and without foam gas inclusions and 4 mm thick were impacted with a constant force from an impact pendulum. Various porosity levels in the foam materials were produced by 1%, 5%, and 10% by weight foaming agent. The forces transmitted through the EVA after energy absorption by the test materials were measured with a force sensor and compared. Results: Only minor non-significant differences in transmitted forces through the EVA with and without foam were shown. Conclusions: The inclusion of gas in the form of a closed cell foam in 4 mm thick EVA mouthguard materials did not improve the impact performance of the EVA mouthguard material.