Global transcriptional responses of the toxic cyanobacterium, Microcystis aeruginosa, to nitrogen stress, phosphorus stress, and growth on organic matter

Whole transcriptome shotgun sequencing (RNA-seq) was used to assess the transcriptomic response of the toxic cyanobacterium Microcystis aeruginosa during growth with low levels of dissolved inorganic nitrogen (low N), low levels of dissolved inorganic phosphorus (low P), and in the presence of high levels of high molecular weight dissolved organic matter (HMWDOM). Under low N, one third of the genome was differentially expressed, with significant increases in transcripts observed among genes within the nir operon, urea transport genes (urtBCDE), and amino acid transporters while significant decreases in transcripts were observed in genes related to photosynthesis. There was also a significant decrease in the transcription of the microcystin synthetase gene set under low N and a significant decrease in microcystin content per Microcystis cell demonstrating that N supply influences cellular toxicity. Under low P, 27% of the genome was differentially expressed. The Pho regulon was induced leading to large increases in transcript levels of the alkaline phosphatase phoX, the Pst transport system (pstABC), and the sphX gene, and transcripts of multiple sulfate transporter were also significantly more abundant. While the transcriptional response to growth on HMWDOM was smaller (5–22% of genes differentially expressed), transcripts of multiple genes specifically associated with the transport and degradation of organic compounds were significantly more abundant within HMWDOM treatments and thus may be recruited by Microcystis to utilize these substrates. Collectively, these findings provide a comprehensive understanding of the nutritional physiology of this toxic, bloom-forming cyanobacterium and the role of N in controlling microcystin synthesis.

Heat-stress and light-stress induce different cellular pathologies in the symbiotic dinoflagellate during coral bleaching

Coral bleaching is a significant contributor to the worldwide degradation of coral reefs and is indicative of the termination of symbiosis between the coral host and its symbiotic algae (dinoflagellate; Symbiodinium sp. complex), usually by expulsion or xenophagy (symbiophagy) of its dinoflagellates. Herein, we provide evidence that during the earliest stages of environmentally induced bleaching, heat stress and light stress generate distinctly different pathomorphological changes in the chloroplasts, while a combined heat- and light-stress exposure induces both pathomorphologies; suggesting that these stressors act on the dinoflagellate by different mechanisms. Within the first 48 hours of a heat stress (32°C) under low-light conditions, heat stress induced decomposition of thylakoid structures before observation of extensive oxidative damage; thus it is the disorganization of the thylakoids that creates the conditions allowing photo-oxidative-stress. Conversely, during the first 48 hours of a light stress (2007 µmoles m−2 s−1 PAR) at 25°C, condensation or fusion of multiple thylakoid lamellae occurred coincidently with levels of oxidative damage products, implying that photo-oxidative stress causes the structural membrane damage within the chloroplasts. Exposure to combined heat- and light-stresses induced both pathomorphologies, confirming that these stressors acted on the dinoflagellate via different mechanisms. Within 72 hours of exposure to heat and/or light stresses, homeostatic processes (e.g., heat-shock protein and anti-oxidant enzyme response) were evident in the remaining intact dinoflagellates, regardless of the initiating stressor. Understanding the sequence of events during bleaching when triggered by different environmental stressors is important for predicting both severity and consequences of coral bleaching

The elastic-plastic indentation response of a columnar thermal barrier coating

Thermal barrier coatings with a columnar microstructure are prone to erosion damage by a mechanism of surface cracking upon impact by small foreign particles. In order to explore this erosion mechanism, the elastic indentation and the elastic-plastic indentation responses of a columnar thermal barrier coating to a spherical indenter were determined by the finite element method and by analytical models. It was shown that the indentation response is intermediate between that of a homogeneous half-space and that given by an elastic-plastic mattress model (with the columns behaving as independent non-linear springs). The sensitivity of the indentation behaviour to geometry and to the material parameters was explored: the diameter of the columns, the gap width between columns, the coefficient of Coulomb friction between columns and the layer height of the thermal barrier coating. The calculations revealed that the level of induced tensile stress is sufficient to lead to cracking of the columns at a depth of about the column radius. It was also demonstrated that the underlying soft bond coat can undergo plastic indentation when the coating comprises parallel columns, but this is less likely for the more realistic case of a random arrangement of tapered columns. © 2009 Elsevier B.V.

Vibration and stress analysis in the presence of structural uncertainty

At medium to high frequencies the dynamic response of a built-up engineering system, such as an automobile, can be sensitive to small random manufacturing imperfections. Ideally the statistics of the system response in the presence of these uncertainties should be computed at the design stage, but in practice this is an extremely difficult task. In this paper a brief review of the methods available for the analysis of systems with uncertainty is presented, and attention is then focused on two particular "non- parametric" methods: statistical energy analysis (SEA), and the hybrid method. The main governing equations are presented, and a number of example applications are considered, ranging from academic benchmark studies to industrial design studies. © 2009 IOP Publishing Ltd.

Development of a universal stress sensor for graphene and carbon fibres

Carbon fibres are a significant volume fraction of modern structural airframes. Embedded into polymer matrices, they provide significant strength and stiffness gains by unit weight compared with competing structural materials. Here we use the Raman G peak to assess the response of carbon fibres to the application of strain, with reference to the response of graphene itself. Our data highlight the predominance of the in-plane graphene properties in all graphitic structures examined. A universal master plot relating the G peak strain sensitivity to tensile modulus of all types of carbon fibres, as well as graphene, is presented. We derive a universal value of - average - phonon shift rate with axial stress of around -5ω0 -1 (cm -1 Mpa-1), where ω0 is the G peak position at zero stress for both graphene and carbon fibre with annular morphology. The use of this for stress measurements in a variety of applications is discussed. © 2011 Macmillan Publishers Limited. All rights reserved.

An analysis of the cooperative mechano-sensitive feedback between intracellular signaling, focal adhesion development, and stress fiber contractility

Cells communicate with their external environment via focal adhesions and generate activation signals that in turn trigger the activity of the intracellular contractile machinery. These signals can be triggered by mechanical loading that gives rise to a cooperative feedback loop among signaling, focal adhesion formation, and cytoskeletal contractility, which in turn equilibrates with the applied mechanical loads. We devise a signaling model that couples stress fiber contractility and mechano-sensitive focal adhesion models to complete this above mentioned feedback loop. The signaling model is based on a biochemical pathway where IP3 molecules are generated when focal adhesions grow. These IP3 molecules diffuse through the cytosol leading to the opening of ion channels that disgorge Ca2+ from the endoplasmic reticulum leading to the activation of the actin/myosin contractile machinery. A simple numerical example is presented where a one-dimensional cell adhered to a rigid substrate is pulled at one end, and the evolution of the stress fiber activation signal, stress fiber concentrations, and focal adhesion distributions are investigated. We demonstrate that while it is sufficient to approximate the activation signal as spatially uniform due to the rapid diffusion of the IP3 through the cytosol, the level of the activation signal is sensitive to the rate of application of the mechanical loads. This suggests that ad hoc signaling models may not be able to capture the mechanical response of cells to a wide range of mechanical loading events. © 2011 American Society of Mechanical Engineers.

Impact response of polyethylene nanocomposites

The quasi-static and dynamic behaviour of Linear Low Density Polyethylene (LLDPE) and two LLDPE nanocomposites were studied. Nanocomposites consisting of LLDPE filled with 1% carbon black and 0.5% nanoclay fillers, by weight, were considered. Under quasi-static tensile loading, an improvement in the energy absorbing capability was achieved by adding 1% carbon black fillers. However, during quasi-static puncture and dynamic impact loading, the advantage provided by the fillers was lost. Thermal softening due to adiabatic heating under high strain rate deformation and difference s in the state of stress are considered as reasons for this reduction. © 2011 Published by Elsevier Ltd.

Dynamic compressive response of composite square honeycombs

Carbon fibre-epoxy composite square honeycombs, and the parent composite material, were tested in quasi-static compression at a strain rate of 10 -3 s -1 and in dynamic compression at strain rates of 10 3-10 4 s -1 using an instrumented Kolsky bar arrangement. Taken together, these tests provide an assessment of the potential of this composite topology for use as a lightweight sandwich core. The honeycombs had two relative densities, 0.12 and 0.24, and two material orientations, ±45° and 0/90° with respect to the prismatic, loading direction of the honeycomb. Honeycomb manufacture was by slotting, assembling and bonding together carbon fibre/epoxy woven plies of composite sheets of 2 × 2 twill weave construction. The peak value of wall stress in the honeycombs was about one third that of the parent material, for all strain rates. An elastic finite element analysis was used to trace the source of this knock-down in strength: a stress concentration exists at the root of the slots and leads to premature failure by microbuckling. Shock-wave effects were evident at impact velocities exceeding 50 ms -1 for the honeycomb of relative density 0.12. This was traced to stubbing of the buckled cell walls against the face of the Kolsky bar. © 2011 Elsevier Ltd. All rights reserved.

Tensile response of passivated films with climb-assisted dislocation glide

The tensile response of single crystal films passivated on two sides is analysed using climb enabled discrete dislocation plasticity. Plastic deformation is modelled through the motion of edge dislocations in an elastic solid with a lattice resistance to dislocation motion, dislocation nucleation, dislocation interaction with obstacles and dislocation annihilation incorporated through a set of constitutive rules. The dislocation motion in the films is by glide-only or by climb-assisted glide whereas in the surface passivation layers dislocation motion occurs by glide-only and penalized by a friction stress. For realistic values of the friction stress, the size dependence of the flow strength of the oxidised films was mainly a geometrical effect resulting from the fact that the ratio of the oxide layer thickness to film thickness increases with decreasing film thickness. However, if the passivation layer was modelled as impenetrable, i.e. an infinite friction stress, the plastic hardening rate of the films increases with decreasing film thickness even for geometrically self-similar specimens. This size dependence is an intrinsic material size effect that occurs because the dislocation pile-up lengths become on the order of the film thickness. Counter-intuitively, the films have a higher flow strength when dislocation motion is driven by climb-assisted glide compared to the case when dislocation motion is glide-only. This occurs because dislocation climb breaks up the dislocation pile-ups that aid dislocations to penetrate the passivation layers. The results also show that the Bauschinger effect in passivated thin films is stronger when dislocation motion is climb-assisted compared to films wherein dislocation motion is by glide-only. © 2012 Elsevier Ltd.

The influence of matrix integrity on stress-fiber remodeling in 3D.

Matrix anisotropy is important for long term in vivo functionality. However, it is not fully understood how to guide matrix anisotropy in vitro. Experiments suggest actin-mediated cell traction contributes. Although F-actin in 2D displays a stretch-avoidance response, 3D data are lacking. We questioned how cyclic stretch influences F-actin and collagen orientation in 3D. Small-scale cell-populated fibrous tissues were statically constrained and/or cyclically stretched with or without biochemical agents. A rectangular array of silicone posts attached to flexible membranes constrained a mixture of cells, collagen I and matrigel. F-actin orientation was quantified using fiber-tracking software, fitted using a bi-model distribution function. F-actin was biaxially distributed with static constraint. Surprisingly, uniaxial cyclic stretch, only induced a strong stretch-avoidance response (alignment perpendicular to stretching) at tissue surfaces and not in the core. Surface alignment was absent when a ROCK-inhibitor was added, but also when tissues were only statically constrained. Stretch-avoidance was also observed in the tissue core upon MMP1-induced matrix perturbation. Further, a strong stretch-avoidance response was obtained for F-actin and collagen, for immediate cyclic stretching, i.e. stretching before polymerization of the collagen. Results suggest that F-actin stress-fibers avoid cyclic stretch in 3D, unless collagen contact guidance dictates otherwise.

The influence of matrix integrity on stress-fiber remodeling in 3D

Matrix anisotropy is important for long term in vivo functionality. However, it is not fully understood how to guide matrix anisotropy in vitro. Experiments suggest actin-mediated cell traction contributes. Although F-actin in 2D displays a stretch-avoidance response, 3D data are lacking. We questioned how cyclic stretch influences F-actin and collagen orientation in 3D. Small-scale cell-populated fibrous tissues were statically constrained and/or cyclically stretched with or without biochemical agents. A rectangular array of silicone posts attached to flexible membranes constrained a mixture of cells, collagen I and matrigel. F-actin orientation was quantified using fiber-tracking software, fitted using a bi-model distribution function. F-actin was biaxially distributed with static constraint. Surprisingly, uniaxial cyclic stretch, only induced a strong stretch-avoidance response (alignment perpendicular to stretching) at tissue surfaces and not in the core. Surface alignment was absent when a ROCK-inhibitor was added, but also when tissues were only statically constrained. Stretch-avoidance was also observed in the tissue core upon MMP1-induced matrix perturbation. Further, a strong stretch-avoidance response was obtained for F-actin and collagen, for immediate cyclic stretching, i.e. stretching before polymerization of the collagen. Results suggest that F-actin stress-fibers avoid cyclic stretch in 3D, unless collagen contact guidance dictates otherwise. © 2012 Elsevier Ltd.

Local stress relaxation and shear banding in a dry foam under shear.

We have developed a realistic simulation of 2D dry foams under quasistatic shear. After a short transient, a shear-banding instability is observed. These results are compared with measurements obtained on real 2D (confined) foams. The numerical model allows us to exhibit the mechanical response of the material to a single plastication event. From the analysis of this elastic propagator, we propose a scenario for the onset and stability of the flow localization process in foams, which should remain valid for most athermal amorphous systems under creep flow.

Factors affecting seismic response of submarine slopes

The response of submerged slopes on the continental shelf to seismic or storm loading has become an important element in the risk assessment for offshore structures and "local" tsunami hazards worldwide. The geological profile of these slopes typically includes normally consolidated to lightly overconsolidated soft cohesive soils with layer thickness ranging from a few meters to hundreds of meters. The factor of safety obtained from pseudo-static analyses is not always a useful measure for evaluating the slope response, since values less than one do not necessarily imply slope failure with large movements of the soil mass. This paper addresses the relative importance of different factors affecting the response of submerged slopes during seismic loading. The analyses use a dynamic finite element code which includes a constitutive law describing the anisotropic stress-strain-strength behavior of normally consolidated to lightly overconsolidated clays. The model also incorporates anisotropic hardening to describe the effect of different shear strain and stress histories as well as bounding surface principles to provide realistic descriptions of the accumulation of the plastic strains and excess pore pressure during successive loading cycles. The paper presents results from parametric site response analyses on slope geometry and layering, soil material parameters, and input ground motion characteristics. The predicted maximum shear strains, permanent deformations, displacement time histories and maximum excess pore pressure development provide insight of slope performance during a seismic event. © 2006 Author(s). This work is licensed under a Creative Commons License.

Seismic response of submarine slopes

The geological profile of submerged slopes on the continental shelf typically includes soft cohesive soils with thicknesses ranging from a few meters to tens or hundreds of meters. The response of these soils in simple shear tests is largely influenced by the presence of an initial consolidation shear stress, inducing anisotropic stress-strain-strength properties which depend also on the direction of shear. In this paper, a new simplified effective-stress-based model describing the behavior of normally to lightly overconsolidated cohesive soils is used in conjunction with a one-dimensional seismic site response analysis computer code to illustrate the importance of accounting for anisotropy and small strain nonlinearity. In particular, a simple example is carried out to compare results for different slope inclinations. Depth profiling of the maximum shear strains and permanent deformations provide insight into the mechanisms of deformation during a seismic event, and the effects of sloping ground conditions.

Compressive response of glass fiber composite sandwich structures

Sandwich panels with crushable foam cores have attracted significant interest for impulsive load mitigation. We describe a method for making a lightweight, energy absorbing, glass fiber composite sandwich structure and explore it is through thickness (out-of-plane) compressive response. The sandwich structure utilized corrugated composite cores constructed from delamination resistant 3D woven E-glass fiber textiles folded over triangular cross section prismatic closed cell, PVC foam inserts. The corrugated structure was stitched to 3D woven S2-glass fiber face sheets and infiltrated with a rubber toughened, impact resistant epoxy. The quasi-static compressive stress-strain response of the panels was experimentally investigated as a function of the strut width to length ratio and compared to micromechanical predictions. Slender struts failed by elastic (Euler) buckling which transitioned to plastic microbuckling as the strut aspect ratio increased. Good agreement was observed between experimental results and micromechanical predictions over the wide range of core densities investigated in the study.