The transcription factor Erg inhibits vascular inflammation by repressing NF-kappaB activation and proinflammatory gene expression in endothelial cells.

OBJECTIVE: To test whether ETS-related gene (Erg) inhibits tumor necrosis factor (TNF)-α-dependent endothelial activation and inflammation. METHODS AND RESULTS: Endothelial activation underlies many vascular diseases, including atherosclerosis. Endothelial activation by proinflammatory cytokines decreases expression of the ETS transcription factor Erg. By using human umbilical vein endothelial cells (HUVECs), we showed that Erg overexpression by adenovirus (AdErg) repressed basal and TNF-α-induced expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), and interleukin 8 (IL-8). Erg inhibited TNF-α-dependent activation of the ICAM-1 promoter, nuclear factor (NF)-κB activity, and NF-κB p65 phosphorylation. Basal NF-κB activity was also inhibited by Erg overexpression. Chromatin immunoprecipitation showed that Erg binds to the ICAM-1 proximal promoter region, which contains 7 putative ETS binding sites. To test the anti-inflammatory role of Erg in vivo, we used a murine model of TNF-α-dependent acute inflammation. The injection of AdErg into the paw decreased TNF-α-induced inflammation compared with control. Finally, staining of human coronary plaques showed loss of Erg expression from the endothelium overlaying active plaque shoulders. CONCLUSIONS: We have identified a novel physiological anti-inflammatory pathway under the control of the transcription factor Erg; this pathway inhibits NF-κB-dependent transcription and TNF-α-induced inflammation in vivo. These results suggest a novel approach to anti-inflammatory therapies.

Laminar shear stress acts as a switch to regulate divergent functions of NF-kappaB in endothelial cells.

Regions of the arterial tree exposed to laminar flow, which exerts high shear stress, are protected from inflammation, endothelial cell (EC) death and atherosclerosis. TNFalpha activates NF-kappaB transcription factors, which potentially exert dual functions by inducing both proinflammatory and cytoprotective transcripts. We assessed whether laminar shear stress protects EC by modulating NF-kappaB function. Human umbilical vein EC (HUVEC) were cultured under shear stress (12 dynes/cm2 for 16 h) using a parallel-plate flow chamber or were maintained in static conditions. Comparative real-time PCR revealed that preshearing significantly alters transcriptional responses to TNFalpha by enhancing the expression of cytoprotective molecules (Bcl-2, MnSOD, GADD45beta, A1) and suppressing proinflammatory transcripts (E-selectin, VCAM-1, IL-8). We demonstrated using assays of nuclear localization, NF-kappaB subunit phosphorylation, DNA-binding, and transcriptional activity that NF-kappaB is activated by TNFalpha in presheared HUVEC. Furthermore, a specific inhibitor revealed that NF-kappaB is essential for the induction of cytoprotective transcripts in presheared EC. Finally, we observed that NF-kappaB can be activated in vascular endothelium exposed to laminar shear stress in NF-kappaB-luciferase reporter mice, thus validating our cell culture experiments. We conclude that shear stress primes EC for enhanced NF-kappaB-dependent cytoprotective responsiveness while attenuating proinflammatory activation. Thus modulation of NF-kappaB function may underlie the atheroprotective effects of laminar shear stress.

A cumulant analysis for non-Gaussian displacement distributions in Newtonian and non-Newtonian flows through porous media.

We use displacement encoding pulsed field gradient (PFG) nuclear magnetic resonance to measure Fourier components S(q) of flow displacement distributions P(zeta) with mean displacement (zeta) for Newtonian and non-Newtonian flows through rocks and bead packs. Displacement distributions are non-Gaussian; hence, there are finite terms above second order in the cumulant expansion of ln(S(q)). We describe an algorithm for an optimal self-consistent cumulant analysis of data, which can be used to obtain the first three (central) moments of a non-Gaussian P(zeta), with error bars. The analysis is applied to Newtonian and non-Newtonian flows in rocks and beads. Flow with shear-thinning xanthan solution produces a 15.6+/-2.3% enhancement of the variance sigma(2) of displacement distributions when compared to flow experiments with water.

Remediation of metal contaminated soil with mineral-amended composts.

This study examined the use of two composts derived from green waste and sewage sludge, amended with minerals (clinoptilolite or bentonite), for the remediation of metal-contaminated brownfield sites to transform them into greenspace. Soils contaminated with high or low levels of metals were mixed with the mineral-enhanced composts at different ratios and assessed by leaching tests, biomass production and metal accumulation of ryegrass (Lolium perenne L.). The results showed that the green waste compost reduced the leaching of Cd and Zn up to 48% whereas the composted sewage sludge doubled the leachate concentration of Zn. However, the same soil amended with composted sewage sludge showed an efficient reduction in plant concentrations of Cd, Cu, Pb or Zn by up to 80%. The results suggest that metal immobilisation and bioavailability are governed by the formation of complexes between the metals and organic matter. The amendment with minerals had only limited effects.

Rapid measurement of transient velocity evolution using GERVAIS.

Rapid velocity measurements using GERVAIS (Gradient Echo Rapid Velocity and Acceleration Imaging Sequence), an EPI (Echo Planar Imaging) based technique capable of measuring velocity over an observation time of several milliseconds, are performed on a wide-gap Couette Rheo-NMR cell for the first time. A variable delay time between a control signal to initiate a transition in flow and the start of the measurement sequence is incorporated to allow investigation of the transient evolution of the velocity field following a step change in rotation rate. Both the commencement and the cessation of imposed shear stress are investigated for (i) a shear banding micellar solution of CPyCl (cetylpyridiniumchloride)/NaSal (sodium salicylate) in brine and (ii) a low molecular weight PDMS (polydimethylsiloxane) oil. With respect to the micellar solution, an elastic shear wave is seen to propagate across the cell following the commencement of shear stress whilst an oscillatory 'recoil' is observed following the cessation of shear stress; neither of these phenomena were observed for the PDMS oil which exhibited a purely viscous response as expected for an incompressible Newtonian fluid. This technique has potential applications across a wide range of transient rheological investigations, particularly with respect to optically opaque materials.