Functional characterization of the initiation enzyme of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus NRS 2004/3a

The glycan chain of the S-layer glycoprotein of Geobacillus stearothermophilus NRS 2004/3a is composed of repeating units [-->2)-alpha-l-Rhap-(1-->3)-beta-l-Rhap-(1-->2)-alpha-l-Rhap-(1-->], with a 2-O-methyl modification of the terminal trisaccharide at the nonreducing end of the glycan chain, a core saccharide composed of two or three alpha-l-rhamnose residues, and a beta-d-galactose residue as a linker to the S-layer protein. In this study, we report the biochemical characterization of WsaP of the S-layer glycosylation gene cluster as a UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase that primes the S-layer glycoprotein glycan biosynthesis of Geobacillus stearothermophilus NRS 2004/3a. Our results demonstrate that the enzyme transfers in vitro a galactose-1-phosphate from UDP-galactose to endogenous phosphoryl-polyprenol and that the C-terminal half of WsaP carries the galactosyltransferase function, as already observed for the UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase WbaP from Salmonella enterica. To confirm the function of the enzyme, we show that WsaP is capable of reconstituting polysaccharide biosynthesis in WbaP-deficient strains of Escherichia coli and Salmonella enterica serovar Typhimurium.

An amino acid substitution in Fasciola hepatica P-glycoprotein from triclabendazole-resistant and triclabendazole-susceptible populations

Control of fasciolosis is threatened by the development of anthelmintic resistance. Enhanced triclabendazole (TCBZ) efflux by ABC transporters such as P-glycoprotein (Pgp) has been implicated in this process. A putative full length cDNA coding for a Pgp expressed in adult Fasciola hepatica has been constructed and used to design a primer set capable of amplifying a region encoding part of the second nucleotide binding domain of Pgp when genomic DNA was used as a template. Application of this primer set to genomic DNA from TCBZ-resistant and -susceptible field populations has shown a significant difference in the alleles present. Analysis of an allele occurring at a three-fold higher frequency in the "resistant" population revealed that it was characterised by a serine to arginine substitution at residue 1144. Homology modelling studies have been used to locate this site in the Pgp structure and hence assess its potential to modify functional activity. © 2012 Elsevier B.V.

Nonlinear Modulated Envelope Electrostatic Wavepacket Propagation in Plasmas

A brief review of the occurrence of amplitude modulated structures in space and laboratory plasmas is provided, followed by a theoretical analysis of the mechanism of carrier wave (self-) interaction, with respect to electrostatic plasma modes. A generic collisionless unmagnetized fluid model is employed. Both cold-(zero-temperature) and warm-(finite temperature) fluid descriptions are considered and compared. The weakly nonlinear oscillation regime is investigated by applying a multiple scale (reductive perturbation) technique and a Nonlinear Schrödinger Equation (NLSE) is obtained, describing the evolution of the slowly varying wave amplitude in time and space. The amplitude’s stability profile reveals the possibility of modulational instability to occur under the influence of external perturbations. The NLSE admits exact localized envelope (solitary wave) solutions of bright (pulses) or dark (holes, voids) type, whose characteristics depend on intrinsic plasma parameters. The role of perturbation obliqueness (with respect to the propagation direction), finite temperature and — possibly — defect (dust) concentration is explicitly considered. The relevance of this description with respect to known electron-ion (e-i) as well as dusty (complex) plasma modes is briefly discussed. © 2004 American Institute of Physics

Opening the climate envelope reveals no macroscale associations with climate in European birds

Predicting how species distributions might shift as global climate changes is fundamental to the successful adaptation of conservation policy. An increasing number of studies have responded to this challenge by using climate envelopes, modeling the association between climate variables and species distributions. However, it is difficult to quantify how well species actually match climate. Here, we use null models to show that species-climate associations found by climate envelope methods are no better than chance for 68 of 100 European bird species. In line with predictions, we demonstrate that the species with distribution limits determined by climate have more northerly ranges. We conclude that scientific studies and climate change adaptation policies based on the indiscriminate use of climate envelope methods irrespective of species sensitivity to climate may be misleading and in need of revision.

Enhanced efficiency of envelope-tracking based broadband inverse class-E power amplifier

In this paper, the envelope-tracking technique is exploited to boost average efficiency of the newly introduced broadband Inverse Class-E power amplifier. A 2.26 GHz - 20.5 dBm - 3 V power amplifier was designed, constructed, and measured. For a multi-carrier input signal with 10 dB peak-to-average ratio, the average PAE was increased from 5.7% to 54.5%. © 2008 IEEE.

Regulatory network of lipopolysaccharide O-antigen biosynthesis in Yersinia enterocolitica includes cell envelope-dependent signals

Lipopolysaccharide (LPS) is a glycolipid present in the outer membrane of all Gram-negative bacteria, and it is one of the signature molecules recognized by the receptors of the innate immune system. In addition to its lipid A portion (the endotoxin), its O-chain polysaccharide (the O-antigen) plays a critical role in the bacterium-host interplay and, in a number of bacterial pathogens, it is a virulence factor. We present evidence that, in Yersinia enterocolitica serotype O:8, a complex signalling network regulates O-antigen expression in response to temperature. Northern blotting and reporter fusion analyses indicated that temperature regulates the O-antigen expression at the transcriptional level. Promoter cloning showed that the O-antigen gene cluster contains two transcriptional units under the control of promoters P(wb1) and P(wb2). The activity of both promoters is under temperature regulation and is repressed in bacteria grown at 37 degrees C. We demonstrate that the RosA/RosB efflux pump/potassium antiporter system and Wzz, the O-antigen chain length determinant, are indirectly involved in the regulation mainly affecting the activity of promoter P(wb2). The rosAB transcription, under the control of P(ros), is activated at 37 degrees C, and P(wb2) is repressed through the signals generated by the RosAB system activation, i.e. decreased [K+] and increased [H+]. The wzz transcription is under the control of P(wb2), and we show that, at 37 degrees C, overexpression of Wzz downregulates slightly the P(wb1) and P(wb2) activities and more strongly the P(ros) activity, with the net result that more O-antigen is produced. Finally, we demonstrate that overexpression of Wzz causes membrane stress that activates the CpxAR two-component signal transduction system.

Dynamical characteristics of solitary waves, shocks and envelope modes in kappa-distributed non-thermal plasmas: an overview

Space plasmas provide abundant evidence of highly energetic particle population, resulting in a long-tailed non-Maxwellian distribution. Furthermore, the first stages in the evolution of plasmas produced during laser-matter interaction are dominated by nonthermal electrons, as confirmed by experimental observation and computer simulations. This phenomenon is efficiently modelled via a kappa-type distribution. We present an overview, from first principles, of the effect of superthermality on the characteristics of electrostatic plasma waves. We rely on a fluid model for ion-acoustic excitations, employing a kappa distribution function to model excess superthermality of the electron distribution. Focusing on nonlinear excitations (solitons), in the form of solitary waves (pulses), shocks and envelope solitons, and employing standard methodological tools of nonlinear plasmadynamical analysis, we discuss the role of excess superthermality in their propagation dynamics (existence laws, stability profile), geometric characteristics and stability. Numerical simulations are employed to confirm theoretical predictions, namely in terms of the stability of electrostatic pulses, as well as the modulational stability profile of bright- and dark-type envelope solitons.

Pushing the Envelope: Future Directions for Evidence-informed Practice

Chapter explores the complexity of implementing evidence-informed practice within the child welfare system in Ontario, Canada. The paper explores the work of Practice and Research Together (PART; www. partcanada.org)