Co-ordination of pathogenicity island expression by the BipA GTPase in enteropathogenic Escherichia coli (EPEC).

BipA is a novel member of the ribosome binding GTPase superfamily and is widely distributed in bacteria and plants. We report here that it regulates -multiple cell surface- and virulence-associated -components in the enteropathogenic Escherichia coli (EPEC) strain E2348/69. The regulated components include bacterial flagella, the espC pathogenicity island and a type III secretion system specified by the locus of enterocyte effacement (LEE). BipA positively regulated the espC and LEE gene clusters through transcriptional control of the LEE-encoded regulator, Ler. Additionally, it affected the pattern of proteolysis of intimin, a key LEE-encoded adhesin specified by the LEE. BipA control of the LEE operated independently of the previously characterized regulators Per, integration host factor and H-NS. In contrast, it negatively regulated the flagella-mediated motility of EPEC and in a Ler-independent manner. Our results indicate that the BipA GTPase functions high up in diverse regulatory cascades to co-ordinate the expression of key pathogenicity islands and other virulence-associated factors in E. coli.

BipA: a tyrosine-phosphorylated GTPase that mediates interactions between enteropathogenic Escherichia coli (EPEC) and epithelial cells.

We report the functional characterization of BipA, a GTPase that undergoes tyrosine phosphorylation in an enteropathogenic Escherichia coli (EPEC) strain. BipA mutants adhere to cultured epithelial cells but fail to trigger the characteristic cytoskeletal rearrangements found in cells infected with wild-type EPEC. In contrast, increased expression of BipA enhances actin remodelling and results in the hyperformation of pseudopods. BipA appears to be the first example of a new class of virulence regulator, as it also controls flagella-mediated cell motility and resistance to the antibacterial effects of a human host defence protein. Its striking sequence similarity to ribosome-binding elongation factors suggests that it uses a novel mechanism to modulate gene expression.

The bacterial cytoskeleton modulates motility, type 3 secretion, and colonization in Salmonella.

Although there have been great advances in our understanding of the bacterial cytoskeleton, major gaps remain in our knowledge of its importance to virulence. In this study we have explored the contribution of the bacterial cytoskeleton to the ability of Salmonella to express and assemble virulence factors and cause disease. The bacterial actin-like protein MreB polymerises into helical filaments and interacts with other cytoskeletal elements including MreC to control cell-shape. As mreB appears to be an essential gene, we have constructed a viable ΔmreC depletion mutant in Salmonella. Using a broad range of independent biochemical, fluorescence and phenotypic screens we provide evidence that the Salmonella pathogenicity island-1 type three secretion system (SPI1-T3SS) and flagella systems are down-regulated in the absence of MreC. In contrast the SPI-2 T3SS appears to remain functional. The phenotypes have been further validated using a chemical genetic approach to disrupt the functionality of MreB. Although the fitness of ΔmreC is reduced in vivo, we observed that this defect does not completely abrogate the ability of Salmonella to cause disease systemically. By forcing on expression of flagella and SPI-1 T3SS in trans with the master regulators FlhDC and HilA, it is clear that the cytoskeleton is dispensable for the assembly of these structures but essential for their expression. As two-component systems are involved in sensing and adapting to environmental and cell surface signals, we have constructed and screened a panel of such mutants and identified the sensor kinase RcsC as a key phenotypic regulator in ΔmreC. Further genetic analysis revealed the importance of the Rcs two-component system in modulating the expression of these virulence factors. Collectively, these results suggest that expression of virulence genes might be directly coordinated with cytoskeletal integrity, and this regulation is mediated by the two-component system sensor kinase RcsC.

Reinforcement learning with reference tracking control in continuous state spaces

The contribution described in this paper is an algorithm for learning nonlinear, reference tracking, control policies given no prior knowledge of the dynamical system and limited interaction with the system through the learning process. Concepts from the field of reinforcement learning, Bayesian statistics and classical control have been brought together in the formulation of this algorithm which can be viewed as a form of indirect self tuning regulator. On the task of reference tracking using a simulated inverted pendulum it was shown to yield generally improved performance on the best controller derived from the standard linear quadratic method using only 30 s of total interaction with the system. Finally, the algorithm was shown to work on the simulated double pendulum proving its ability to solve nontrivial control tasks. © 2011 IEEE.

Designing and testing an advanced pneumatic braking system for heavy vehicles

Heavy goods vehicles exhibit poor braking performance in emergency situations when compared to other vehicles. Part of the problem is caused by sluggish pneumatic brake actuators, which limit the control bandwidth of their antilock braking systems. In addition, heuristic control algorithms are used that do not achieve the maximum braking force throughout the stop. In this article, a novel braking system is introduced for pneumatically braked heavy goods vehicles. The conventional brake actuators are improved by placing high-bandwidth, binary-actuated valves directly on the brake chambers. A made-for-purpose valve is described. It achieves a switching delay of 3-4 ms in tests, which is an order of magnitude faster than solenoids in conventional anti-lock braking systems. The heuristic braking control algorithms are replaced with a wheel slip regulator based on sliding mode control. The combined actuator and slip controller are shown to reduce stopping distances on smooth and rough, high friction (μ = 0.9) surfaces by 10% and 27% respectively in hardware-in-the-loop tests compared with conventional ABS. On smooth and rough, low friction (μ = 0.2) surfaces, stopping distances are reduced by 23% and 25%, respectively. Moreover, the overall air reservoir size required on a heavy goods vehicle is governed by its air usage during an anti-lock braking stop on a low friction, smooth surface. The 37% reduction in air usage observed in hardware-in-the-loop tests on this surface therefore represents the potential reduction in reservoir size that could be achieved by the new system. © 2012 IMechE.

Adaptive combustion instability control with saturation

The control of a class of combustion systems, suceptible to damage from self-excited combustion oscillations, is considered. An adaptive stable controller, called Self-Tuning Regulator (STR), has recently been developed, which meets the apparently contradictory challenge of relying as little as possible on a particular combustion model while providing some guarantee that the controller will cause no harm. The controller injects some fuel unsteadily into the burning region, thereby altering the heat release, in response to an input signal detecting the oscillation. This paper focuses on an extension of the STR design, when, due to stringent emission requirements and to the danger of flame extension, the amount of fuel used for control is limited in amplitude. A Lyapunov stability analysis is used to prove the stability of the modified STR when the saturation constraint is imposed. The practical implementation of the modified STR remains straightforward, and simulation results, based on the nonlinear premixed flame model developed by Dowling, show that in the presence of a saturation constraint, the self-excited oscillations are damped more rapidly with the modified STR than with the original STR. © 2001 by S. Evesque. Published by the American Institute of Aeronautics and Astronautics, Inc.

The adoption of sustainable remediation behaviour in the US and UK: a cross country comparison and determinant analysis.

The sustainable remediation concept, aimed at maximizing the net environmental, social, and economic benefits in contaminated site remediation, is being increasingly recognized by industry, governments, and academia. However, there is limited understanding of actual sustainable behaviour being adopted and the determinants of such sustainable behaviour. The present study identified 27 sustainable practices in remediation. An online questionnaire survey was used to rank and compare them in the US (n=112) and the UK (n=54). The study also rated ten promoting factors, nine barriers, and 17 types of stakeholders' influences. Subsequently, factor analysis and general linear models were used to determine the effects of internal characteristics (i.e. country, organizational characteristics, professional role, personal experience and belief) and external forces (i.e. promoting factors, barriers, and stakeholder influences). It was found that US and UK practitioners adopted many sustainable practices to similar extents. Both US and UK practitioners perceived the most effectively adopted sustainable practices to be reducing the risk to site workers, protecting groundwater and surface water, and reducing the risk to the local community. Comparing the two countries, we found that the US adopted innovative in-situ remediation more effectively; while the UK adopted reuse, recycling, and minimizing material usage more effectively. As for the overall determinants of sustainable remediation, the country of origin was found not to be a significant determinant. Instead, organizational policy was found to be the most important internal characteristic. It had a significant positive effect on reducing distant environmental impact, sustainable resource usage, and reducing remediation cost and time (p<0.01). Customer competitive pressure was found to be the most extensively significant external force. In comparison, perceived stakeholder influence, especially that of primary stakeholders (site owner, regulator, and primary consultant), did not appear to have as extensive a correlation with the adoption of sustainability as one would expect.