Autoregulation of the Escherichia coli melR promoter: repression involves four molecules of MelR

The Escherichia coli MelR protein is a transcription activator that autoregulates its own promoter by repressing transcription initiation. Optimal repression requires MelR binding to a site that overlaps the melR transcription start point and to upstream sites. In this work, we have investigated the different determinants needed for optimal repression and their spatial requirements. We show that repression requires a complex involving four DNA-bound MelR molecules, and that the global CRP regulator plays little or no role

Dopamine D2-like receptor stimulation blocks negative feedback in visual and spatial reversal learning in the rat: behavioural and computational evidence

Abstract: Rationale: Dopamine D2-like receptors (D2R) are important drug targets in schizophrenia and Parkinson’s disease, but D2R ligands also cause cognitive inflexibility such as poor reversal learning. The specific role of D2R in reversal learning remains unclear. Objectives: We tested the hypotheses that D2R agonism impairs reversal learning by blocking negative feedback and that antagonism of D1-like receptors (D1R) impairs learning from positive feedback. Methods: Male Lister Hooded rats were trained on a novel visual reversal learning task. Performance on “probe trials”, during which the correct or incorrect stimulus was presented with a third, probabilistically rewarded (50% of trials) and therefore intermediate stimulus, revealed individual learning curves for the processes of positive and negative feedback. The effects of D2R and D1R agonists and antagonists were evaluated. A separate cohort was tested on a spatial probabilistic reversal learning (PRL) task after D2R agonism. Computational reinforcement learning modelling was applied to choice data from the PRL task to evaluate the contribution of latent factors. Results: D2R agonism with quinpirole dose-dependently impaired both visual reversal and PRL. Analysis of the probe trials on the visual task revealed a complete blockade of learning from negative feedback at the 0.25 mg/kg dose, while learning from positive feedback was intact. Estimated parameters from the model that best described the PRL choice data revealed a steep and selective decrease in learning rate from losses. D1R antagonism had a transient effect on the positive probe trials. Conclusions: D2R stimulation impairs reversal learning by blocking the impact of negative feedback

Comparative analyses imply that the enigmatic sigma factor 54 is a central controller of the bacterial exterior

Contains fulltext : 95738.pdf (publisher's version ) (Open Access)BACKGROUND: Sigma-54 is a central regulator in many pathogenic bacteria and has been linked to a multitude of cellular processes like nitrogen assimilation and important functional traits such as motility, virulence, and biofilm formation. Until now it has remained obscure whether these phenomena and the control by Sigma-54 share an underlying theme. RESULTS: We have uncovered the commonality by performing a range of comparative genome analyses. A) The presence of Sigma-54 and its associated activators was determined for all sequenced prokaryotes. We observed a phylum-dependent distribution that is suggestive of an evolutionary relationship between Sigma-54 and lipopolysaccharide and flagellar biosynthesis. B) All Sigma-54 activators were identified and annotated. The relation with phosphotransfer-mediated signaling (TCS and PTS) and the transport and assimilation of carboxylates and nitrogen containing metabolites was substantiated. C) The function annotations, that were represented within the genomic context of all genes encoding Sigma-54, its activators and its promoters, were analyzed for intra-phylum representation and inter-phylum conservation. Promoters were localized using a straightforward scoring strategy that was formulated to identify similar motifs. We found clear highly-represented and conserved genetic associations with genes that concern the transport and biosynthesis of the metabolic intermediates of exopolysaccharides, flagella, lipids, lipopolysaccharides, lipoproteins and peptidoglycan. CONCLUSION: Our analyses directly implicate Sigma-54 as a central player in the control over the processes that involve the physical interaction of an organism with its environment like in the colonization of a host (virulence) or the formation of biofilm

Understanding glucose transport by the bacterial phosphoenolpyruvate. Glycose phosphotransferase system on the basis of kinetic measurements in vitro.

The kinetic parameters in vitro of the components of the phosphoenolpyruvate:glycose phosphotransferase system (PTS) in enteric bacteria were collected. To address the issue of whether the behavior in vivo of the PTS can be understood in terms of these enzyme kinetics, a detailed kinetic model was constructed. Each overall phosphotransfer reaction was separated into two elementary reactions, the first entailing association of the phosphoryl donor and acceptor into a complex and the second entailing dissociation of the complex into dephosphorylated donor and phosphorylated acceptor. Literature data on the K(m) values and association constants of PTS proteins for their substrates, as well as equilibrium and rate constants for the overall phosphotransfer reactions, were related to the rate constants of the elementary steps in a set of equations; the rate constants could be calculated by solving these equations simultaneously. No kinetic parameters were fitted. As calculated by the model, the kinetic parameter values in vitro could describe experimental results in vivo when varying each of the PTS protein concentrations individually while keeping the other protein concentrations constant. Using the same kinetic constants, but adjusting the protein concentrations in the model to those present in cell-free extracts, the model could reproduce experiments in vitro analyzing the dependence of the flux on the total PTS protein concentration. For modeling conditions in vivo it was crucial that the PTS protein concentrations be implemented at their high in vivo values. The model suggests a new interpretation of results hitherto not understood; in vivo, the major fraction of the PTS proteins may exist as complexes with other PTS proteins or boundary metabolites, whereas in vitro, the fraction of complexed proteins is much smaller

New avenues for failure analysis

As evidenced by the presentations at ICEFA-3, failure in electronics equipment has received little attention from the Failure Analysis Community. Electronics is the World’s largest industrial sector, fiercely competitive and facing challenges from environmental demands and continuous miniaturisation. The paper outlines the basic elements in electronic equipment, typical operating conditions, common defects and the likely modes of failure in service. It demonstrates the greater resilience required by solders than most traditional alloys at their peak operating temperatures. With rapidly changing technologies, employment of new alloy systems and dimensions such that properties can no longer be described by bulk measurements, it is contended that the scope for failure and failure analysis is significant and growing

Defects in electronics and their significance for structural integrity

With Electronics likely to become a major area of interest to structural integrity practitioners, the paper aims to introduce the field from a structural integrity perspective. The Industry is presently facing two major challenges: continued miniaturisation and the implementation of lead-free technology associated with interconnections. Following a consideration of the background of both aspects, a brief outline of typical components and production processes is presented. Attention is then focussed upon possible defects that may arise during manufacture, and under the, often, complex conditions of service. Comparisons are drawn with more traditional applications of structural integrity, and the fresh challenges associated with the utilisation of a new generation of solder alloys, and with micro and near-nano dimensions, are introduced

Damage Produced in Solder Alloys during Thermal Cycling

The anisotropy of tin is associated with significant variations in its coefficient of thermal expansion and elastic modulus, with crystallographic direction. Under pure thermal cycling (with no externally applied stress or strain), substantial strains, in excess of 100%, may develop locally, and for very small structures, such as soldered interconnections comprising a few grains, structural integrity may be adversely affected. To examine this possibility, freestanding samples of tin, Sn-3.5wt.%Ag, Sn-0.5wt.%Cu, and Sn-3.8wt.%Ag-0.7wt.%Cu, have been subjected to thermal cycling. Temperature cycles from 30 degrees C to 125 degrees C or from -40 degrees C to 55 degrees C initially caused surface cracking, with openings up to several tens of microns after 3,000 cycles. Subsequently, the surface cracks grew into the interior of the specimens, with the maximum penetration ranging from a few microns after 100 cycles to more than 200 pm after 3,000 cycles. The cracks initiated from damage accumulated along grain boundaries. For the same temperature range, less damage resulted after the lower maximum (or mean) temperature cycle, and there appears to be a thermally activated component of cracking. The microstructure produced by rapid cooling (water quenching) was slightly more resistant than that formed by air, or furnace, cooling. Apart from microstructural coarsening, no damage accrues from isothermal exposure alone

Effects of strain rate and temperature on the stress–strain response of solder alloys

To ensure reliable design of soldered interconnections as electronic devices become smaller, requires greater knowledge and understanding of the relevant mechanical behavior of solder alloys than are presently available. The present paper reports the findings of an investigation into the monotonic tensile properties of bulk samples of three solder alloys; a lead–tin eutectic and two lead-free solders (tin–3.5 copper and a tin–3.5 silver alloy). Temperatures between–10 and 75°C and strain rates between 10–1 and 10–3 s–1 have been studied. Both temperature and strain rate may have a substantial effect on strength, producing changes well in excess of 100%. Strength is reduced by lowering strain rate and increasing temperature, and Sn–37 Pb is usually most sensitive to the latter. Expressions for strain and strain rate hardening have been developed. The Sn–0.5 Cu alloy is usually the weakest and most ductile. Sn–37 Pb is strongest at room temperature but with increasing temperature and lower strain rates it becomes inferior to Sn–3.5 Ag. Ductility changes with temperature and strain rate for all three alloys are generally small with inconsistent trends. The role of such data in stress analysis and modeling is considered and the paramount importance of employing data for conditions appropriate to service, is emphasized