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.

High order schemes on three-dimensional general polyhedral meshes - Application to the level set method

In this article, we detail the methodology developed to construct arbitrarily high order schemes - linear and WENO - on 3D mixed-element unstructured meshes made up of general convex polyhedral elements. The approach is tailored specifically for the solution of scalar level set equations for application to incompressible two-phase flow problems. The construction of WENO schemes on 3D unstructured meshes is notoriously difficult, as it involves a much higher level of complexity than 2D approaches. This due to the multiplicity of geometrical considerations introduced by the extra dimension, especially on mixed-element meshes. Therefore, we have specifically developed a number of algorithms to handle mixed-element meshes composed of convex polyhedra with convex polygonal faces. The contribution of this work concerns several areas of interest: the formulation of an improved methodology in 3D, the minimisation of computational runtime in the implementation through the maximum use of pre-processing operations, the generation of novel methods to handle complex 3D mixed-element meshes and finally the application of the method to the transport of a scalar level set. © 2012 Global-Science Press.

Autoignition of monodisperse biodiesel and diesel sprays in turbulent flows

A new experimental configuration has been developed to examine the effects of flow on the autoignition of dilute diesel and biodiesel sprays, where the spray is injected in the form of monodisperse individual droplets at right angles to a hot air turbulent flow. The ignition location has been measured by monitoring the OH * chemiluminescence. A qualitative comparison of the flame behaviour between ethanol, acetone, heptane and biodiesel as fuels has also been carried out. With decreasing volatility of the fuel, the flame showed progressively a higher number of individual droplets burning, with the first autoignition spots appearing at random locations but in general earlier than the intense droplet-flame emission. The time-averaged autoignition length increased with increasing air velocity and with increasing intensity of the turbulence, while it decreased with the temperature and the droplet size. The data can be used for validating models for two-phase turbulent combustion. © 2012 Elsevier Inc.

Characteristics of blood flow in microchannels and relevant impact on modelling blood behaviour in biochip separators

This paper reports a perspective investigation of computational modelling of blood fluid in microchannel devices as a preparation for future research on fluid-structure interaction (FSI) in biofluid mechanics. The investigation is carried out through two aspects, respectively on physical behaviours of blood flow in microchannels and appropriate methodology for modelling. The physics of blood flow is targeted to the challenges for describing blood flow in microchannels, including rheology of blood fluid, suspension features of red blood cells (RBCs), laminar hydrodynamic influence and effect of surface roughness. The analysis shows that due to the hyperelastic property of RBC and its comparable dimension with microchannels, blood fluid shows complex behaviours of two phase flow. The trajectory and migration of RBCs require accurate description of RBC deformation and interaction with plasma. Following on a discussion of modelling approaches, i.e. Eulerian method and Lagrangian method, the main stream modelling methods for multiphase flow are reviewed and their suitability to blood flow is analysed. It is concluded that the key issue for blood flow modelling is how to describe the suspended blood cells, modelled by Lagrangian method, and couple them with the based flow, modelled by Eulerian method. The multiphase flow methods are thereby classified based on the number of points required for describing a particle, as follows: (i) single-point particle methods, (ii) mutli-point particle methods, (iii) functional particle methods, and (iv) fluid particle methods. While single-point particle methods concentrate on particle dynamic movement, multipoint and functional particle methods can take into account particle mechanics and thus offer more detailed information for individual particles. Fluid particle methods provide good compromise between two phases, but require additional information for particle mechanics. For furthermore detailed description, we suggest to investigate the possibility using two domain coupling method, in which particles and base flow are modelled by two separated solvers. It is expected that this paper could clarify relevant issues in numerical modelling of blood flow in microchannels and induce some considerations for modelling blood flow using multiphase flow methods. © 2012 IEEE.

Multi-scale modelling of granular avalanches

Avalanches, debris flows, and landslides are geophysical hazards, which involve rapid mass movement of granular solids, water and air as a single-phase system. The dynamics of a granular flow involve at least three distinct scales: the micro-scale, meso-scale, and the macro-scale. This study aims to understand the ability of continuum models to capture the micro-mechanics of dry granular collapse. Material Point Method (MPM), a hybrid Lagrangian and Eulerian approach, with Mohr-Coulomb failure criterion is used to describe the continuum behaviour of granular column collapse, while the micromechanics is captured using Discrete Element Method (DEM) with tangential contact force model. The run-out profile predicted by the continuum simulations matches with DEM simulations for columns with small aspect ratios ('h/r' < 2), however MPM predicts larger run-out distances for columns with higher aspect ratios ('h/r' > 2). Energy evolution studies in DEM simulations reveal higher collisional dissipation in the initial free-fall regime for tall columns. The lack of a collisional energy dissipation mechanism in MPM simulations results in larger run-out distances. Micro-structural effects, such as shear band formations, were observed both in DEM and MPM simulations. A sliding flow regime is observed above the distinct passive zone at the core of the column. Velocity profiles obtained from both the scales are compared to understand the reason for a slow flow run-out mobilization in MPM simulations. © 2013 AIP Publishing LLC.

On kinematic relaxation and deposition of water droplets in the last stages of low pressure steam turbines

In the first part of the paper steady two-phase flow predictions have been performed for the last stage of a model steam turbine to examine the influence of drag between condensed fog droplets and the continuous vapour phase. In general, droplets due to homogeneous condensation are small and thus kinematic relaxation provides only a minor contribution to the wetness losses. Different droplet size distributions have been investigated to estimate at which size inter-phase friction becomes more important. The second part of the paper deals with the deposition of fog droplets on stator blades. Results from several references are repeated to introduce the two main deposition mechanisms which are inertia and turbulent diffusion. Extensive postprocessing routines have been programmed to calculate droplet deposition due to these effects for a last stage stator blade in three-dimensions. In principle the method to determine droplet deposition by turbulent diffusion equates to that of Yau and Young [1] and the advantages and disadvantages of this relatively simple method are discussed. The investigation includes the influence of different droplet sizes on droplet deposition rates and shows that for small fog droplets turbulent diffusion is the main deposition mechanism. If the droplets size is increased inertial effects become more and more important and for droplets around 1 μm inertial deposition dominates. Assuming realistic droplet sizes the overall deposition equates to about 1% to 3% of the incoming wetness for the investigated guide vane at normal operating conditions. Copyright © 2013 by Solar Turbines Incorporated.

Water droplet flow paths and droplet deposition in low pressure steam turbines

The complex three-dimensional two-phase flow in a low pressure steam turbine is investigated with comprehensive numerical flow simulations. In addition to the condensation process, which already takes place in the last stages of steam turbines, the numerical flow model is enhanced to consider the drag forces between the droplets and the vapour phase. The present paper shows the differences in the flow path of the phases and investigates the effect of an increasing droplet diameter. For the flow simulations a performance cluster is used because of the high effort for such multi-momentum two-phase flow calculations. In steam turbines the deposition of small water droplets on the stator blades or on parts of the casing is responsible for the formation of large coarse water droplets and these may cause additional dissipation as well as damage due to blade erosion. A method is presented that uses detailed CFD data to predict droplet deposition on turbine stator blades. This simulation method to detect regions of droplet deposition can help to improve the design of water removal devices. © Springer-Verlag Berlin Heidelberg 2013.

Unsteady numerical study of wet steam flow in a low pressure steam turbine

In steam power plants condensation already starts in the flow path of the low pressure part of the steam turbine, which leads to a complex three-dimensional two-phase flow. Wetness losses are caused due to thermodynamic and mechanical relaxation processes during condensation and droplet transport. The present investigation focuses on the unsteady effects due to rotor-stator interaction on the droplet formation process. Results of unsteady three dimensional flow simulations of a two-stage steam turbine are presented, whereby this is the first time that non-equilibrium condensation is considered in such simulations. The numerical approach is based on RANS equations, which are extended by a wet steam specific nucleation and droplet growth model. Despite the use of a high performance cluster the unsteady simulation has a considerably high simulation time of approximately 60 days by use of 48 CPUs. © Springer-Verlag Berlin Heidelberg 2012.

Wetness loss prediction for a low pressure steam turbine using computational fluid dynamics

Two-phase computational fluid dynamics modelling is used to investigate the magnitude of different contributions to the wet steam losses in a three-stage model low pressure steam turbine. The thermodynamic losses (due to irreversible heat transfer across a finite temperature difference) and the kinematic relaxation losses (due to the frictional drag of the drops) are evaluated directly from the computational fluid dynamics simulation using a concept based on entropy production rates. The braking losses (due to the impact of large drops on the rotor) are investigated by a separate numerical prediction. The simulations show that in the present case, the dominant effect is the thermodynamic loss that accounts for over 90% of the wetness losses and that both the thermodynamic and the kinematic relaxation losses depend on the droplet diameter. The numerical results are brought into context with the well-known Baumann correlation, and a comparison with available measurement data in the literature is given. The ability of the numerical approach to predict the main wetness losses is confirmed, which permits the use of computational fluid dynamics for further studies on wetness loss correlations. © IMechE 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Rapid patterning of 1-D collagenous topography as an ECM protein fibril platform for image cytometry.

Cellular behavior is strongly influenced by the architecture and pattern of its interfacing extracellular matrix (ECM). For an artificial culture system which could eventually benefit the translation of scientific findings into therapeutic development, the system should capture the key characteristics of a physiological microenvironment. At the same time, it should also enable standardized, high throughput data acquisition. Since an ECM is composed of different fibrous proteins, studying cellular interaction with individual fibrils will be of physiological relevance. In this study, we employ near-field electrospinning to create ordered patterns of collagenous fibrils of gelatin, based on an acetic acid and ethyl acetate aqueous co-solvent system. Tunable conformations of micro-fibrils were directly deposited onto soft polymeric substrates in a single step. We observe that global topographical features of straight lines, beads-on-strings, and curls are dictated by solution conductivity; whereas the finer details such as the fiber cross-sectional profile are tuned by solution viscosity. Using these fibril constructs as cellular assays, we study EA.hy926 endothelial cells' response to ROCK inhibition, because of ROCK's key role in the regulation of cell shape. The fibril array was shown to modulate the cellular morphology towards a pre-capillary cord-like phenotype, which was otherwise not observed on a flat 2-D substrate. Further facilitated by quantitative analysis of morphological parameters, the fibril platform also provides better dissection in the cells' response to a H1152 ROCK inhibitor. In conclusion, the near-field electrospun fibril constructs provide a more physiologically-relevant platform compared to a featureless 2-D surface, and simultaneously permit statistical single-cell image cytometry using conventional microscopy systems. The patterning approach described here is also expected to form the basics for depositing other protein fibrils, seen among potential applications as culture platforms for drug screening.