Inhibition of iron-catalysed hydroxyl radical formation by inositol polyphosphates: a possible physiological function for myo-inositol hexakisphosphate

1. The ability of myo-inositol polyphosphates to inhibit iron-catalysed hydroxyl radical formation was studied in a hypoxanthine/xanthine oxidase system [Graf, Empson and Eaton (1987) J. Biol. Chem. 262, 11647-11650]. Fe3+ present in the assay reagents supported some radical formation, and a standard assay, with 5 microM Fe3+ added, was used to investigate the specificity of compounds which could inhibit radical generation. 2. InsP6 (phytic acid) was able to inhibit radical formation in this assay completely. In this respect it was similar to the effects of the high affinity Fe3+ chelator Desferral, and dissimilar to the effects of EDTA which, even at high concentrations, still allowed detectable radical formation to take place. 3. The six isomers of InsP5 were purified from an alkaline hydrolysate of InsP6 (four of them as two enantiomeric mixtures) and they were compared with InsP6 in this assay. Ins(1,2,3,4,6)P5 and D/L-Ins(1,2,3,4,5)P5 were similar to InsP6 in that they caused a complete inhibition of iron-catalysed radical formation at > 30 microM. Ins(1,3,4,5,6)P5 and D/L-Ins(1,2,4,5,6)P5, however, were markedly less potent than InsP6, and did not inhibit radical formation completely; even when Ins(1,3,4,5,6)P5 was added up to 600 microM, significant radical formation was still detected. Thus InsP5s lacking 2 or 1/3 phosphates are in this respect qualitatively different from InsP6 and the other InsP5s. 4. scyllo-Inositol hexakisphosphate was also tested, and although it caused a greater inhibition than Ins(1,3,4,5,6)P5, it too still allowed detectable free radical formation even at 600 microM. 5. We conclude that the 1,2,3 (equatorial-axial-equatorial) phosphate grouping in InsP6 has a conformation that uniquely provides a specific interaction with iron to inhibit totally its ability to catalyse hydroxyl radical formation; we suggest that a physiological function of InsP6 might be to act as a 'safe' binding site for iron during its transport through the cytosol or cellular organelles

Measuring performance of virtual learning environment system in higher education

Purpose – The purpose of this paper is to measure the performance of commercial virtual learning environment (VLE) systems, which helps the decision makers to select the appropriate system for their institutions. Design/methodology/approach – This paper develops an integrated multiple criteria decision making approach, which combines the analytic hierarchy process (AHP) and quality function deployment (QFD), to evaluate and select the best system. The evaluating criteria are derived from the requirements of those who use the system. A case study is provided to demonstrate how the integrated approach works. Findings – The major advantage of the integrated approach is that the evaluating criteria are of interest to the stakeholders. This ensures that the selected system will achieve the requirements and satisfy the stakeholders most. Another advantage is that the approach can guarantee the benchmarking to be consistent and reliable. From the case study, it is proved that the performance of a VLE system being used at the university is the best. Therefore, the university should continue to run the system in order to support and facilitate both teaching and learning. Originality/value – It is believed that there is no study that measures the performance of VLE systems, and thus decision makers may have difficulties in system evaluation and selection for their institutions.

Dynamics of on-line learning in radial basis function networks

On-line learning is examined for the radial basis function network, an important and practical type of neural network. The evolution of generalization error is calculated within a framework which allows the phenomena of the learning process, such as the specialization of the hidden units, to be analyzed. The distinct stages of training are elucidated, and the role of the learning rate described. The three most important stages of training, the symmetric phase, the symmetry-breaking phase, and the convergence phase, are analyzed in detail; the convergence phase analysis allows derivation of maximal and optimal learning rates. As well as finding the evolution of the mean system parameters, the variances of these parameters are derived and shown to be typically small. Finally, the analytic results are strongly confirmed by simulations.

Analysis of tissue transglutaminase function in the migration of Swiss 3T3 fibroblasts:the active-state conformation of the enzyme does not affect cell motility but is important for its secretion

Increasing evidence suggests that tissue transglutaminase (tTGase; type II) is externalized from cells, where it may play a key role in cell attachment and spreading and in the stabilization of the extracellular matrix (ECM) through protein cross-linking. However, the relationship between these different functions and the enzyme's mechanism of secretion is not fully understood. We have investigated the role of tTGase in cell migration using two stably transfected fibroblast cell lines in which expression of tTGase in its active and inactive (C277S mutant) states is inducible through the tetracycline-regulated system. Cells overexpressing both forms of tTGase showed increased cell attachment and decreased cell migration on fibronectin. Both forms of the enzyme could be detected on the cell surface, but only the clone overexpressing catalytically active tTGase deposited the enzyme into the ECM and cell growth medium. Cells overexpressing the inactive form of tTGase did not deposit the enzyme into the ECM or secrete it into the cell culture medium. Similar results were obtained when cells were transfected with tTGase mutated at Tyr(274) (Y274A), the proposed site for the cis,trans peptide bond, suggesting that tTGase activity and/or its tertiary conformation dependent on this bond may be essential for its externalization mechanism. These results indicate that tTGase regulates cell motility as a novel cell-surface adhesion protein rather than as a matrix-cross-linking enzyme. They also provide further important insights into the mechanism of externalization of the enzyme into the extracellular matrix.

A type system with usage aspects

Linear typing schemes can be used to guarantee non-interference and so the soundness of in-place update with respect to a functional semantics. But linear schemes are restrictive in practice, and more restrictive than necessary to guarantee soundness of in-place update. This limitation has prompted research into static analysis and more sophisticated typing disciplines to determine when in-place update may be safely used, or to combine linear and non-linear schemes. Here we contribute to this direction by defining a new typing scheme that better approximates the semantic property of soundness of in-place update for a functional semantics. We begin from the observation that some data are used only in a read-only context, after which it may be safely re-used before being destroyed. Formalising the in-place update interpretation in a machine model semantics allows us to refine this observation, motivating three usage aspects apparent from the semantics that are used to annotate function argument types. The aspects are (1) used destructively, (2), used read-only but shared with result, and (3) used read-only and not shared with the result. The main novelty is aspect (2), which allows a linear value to be safely read and even aliased with a result of a function without being consumed. This novelty makes our type system more expressive than previous systems for functional languages in the literature. The system remains simple and intuitive, but it enjoys a strong soundness property whose proof is non-trivial. Moreover, our analysis features principal types and feasible type reconstruction, as shown in M. Konen'y (In TYPES 2002 workshop, Nijmegen, Proceedings, Springer-Verlag, 2003).