Effect of site-specific mutations in different phosphotransfer domains of the chemosensory protein ChpA on Pseudomonas aeruginosa motility

The virulence of Pseudomonas aeruginosa and other surface pathogens involves the coordinate expression of a wide range of virulence determinants, including type IV pili. These surface filaments are important for the colonization of host epithelial tissues and mediate bacterial attachment to, and translocation across, surfaces by a process known as twitching motility. This process is controlled in part by a complex signal transduction system whose central component, ChpA, possesses nine potential sites of phosphorylation, including six histidine-containing phosphotransfer (HPt) domains, one serine-containing phosphotransfer domain, one threonine-containing phosphotransfer domain, and one CheY-like receiver domain. Here, using site-directed mutagenesis, we show that normal twitching motility is entirely dependent on the CheY-like receiver domain and partially dependent on two of the HPt domains. Moreover, under different assay conditions, point mutations in several of the phosphotransfer domains of ChpA give rise to unusual "swarming" phenotypes, possibly reflecting more subtle perturbations in the control of P. aeruginosa motility that are not evident from the conventional twitching stab assay. Together, these results suggest that ChpA plays a central role in the complex regulation of type IV pilus-mediated motility in P. aeruginos

Sialic Acid Mutarotation Is Catalyzed by the Escherichia coli β-Propeller Protein YjhT

The acquisition of host-derived sialic acid is an important virulence factor for some bacterial pathogens, but in vivo this sugar acid is sequestered in sialoconjugates as the {alpha}-anomer. In solution, however, sialic acid is present mainly as the β-anomer, formed by a slow spontaneous mutarotation. We studied the Escherichia coli protein YjhT as a member of a family of uncharacterized proteins present in many sialic acid-utilizing pathogens. This protein is able to accelerate the equilibration of the {alpha}- and β-anomers of the sialic acid N-acetylneuraminic acid, thus describing a novel sialic acid mutarotase activity. The structure of this periplasmic protein, solved to 1.5Å resolution, reveals a dimeric 6-bladed unclosed β-propeller, the first of a bacterial Kelch domain protein. Mutagenesis of conserved residues in YjhT demonstrated an important role for Glu-209 and Arg-215 in mutarotase activity. We also present data suggesting that the ability to utilize {alpha}-N-acetylneuraminic acid released from complex sialoconjugates in vivo provides a physiological advantage to bacteria containing YjhT

A study of the deformation of helical springs under eccentric loading

Much analysis has been done to date on the deformation of helical springs due to normal loadings. The aim of this study is to design a helical spring that will deform under eccentric loading a desired amount due to a given force. Under the assumptions of linear stress strain relationships, the spring will be designed in terms of its material properties and its geometry. The deformation of the spring will be made possible utilizing a Shape Memory Alloy (SMA) active element that undergoes phase transformation upon heating above a certain temperature. Two models for spring deformation have been considered. In the first model we study the differential compression of a spring using SMA wire actuators, and in the second model we investigate the bending of an SMA rod placed inside the spring. Our efforts were a first step towards the development of a structural skeleton for a minimally invasive surgical manipulatorhttp://archive.org/details/studyofdeformati00leecLieutenant, United States NavyApproved for public release; distribution is unlimited

Cluster Populations in A115 and A2283

This paper presents four color narrow-band photometry of clusters A115 ($z=0.191$) and A2283 ($z=0.182$) in order to follow the star formation history of various galaxy types. Although located at similar redshifts, the two clusters display very different fractions of blue galaxies (i.e. the Butcher-Oemler effect, $f_B = 0.13$ for A115, $f_B = 0.30$ for A2283). A system of photometric classification is applied to the cluster members that divides the cluster population into four classes based on their recent levels of star formation. It is shown that the blue population of each cluster is primarily composed of normal starforming (SFR < 1 M_{\sun} yrs$^{-1}$) galaxies at the high luminosity end, but with an increasing contribution from a dwarf starburst population below $M_{5500}= -20$. This dwarf starburst population appears to be the same population of low mass galaxies identified in recent HST imaging (Koo et al 1997), possible progenitors to present-day cluster dwarf ellipticals, irregulars and BCD's. Deviations in the color-magnitude relationship for the red galaxies in each cluster suggest that a population of blue S0's is evolving into present-day S0 colors at this epoch. The radial distribution of the blue population supports the prediction of galaxy harassment mechanisms for tidally induced star formation operating on an infalling set of gas-rich galaxies.Comment: 28 pages including 2 tables and 9 figures, AASTeX v4.0. Accepted by Ap.J. Data, referee report and response are avaliable from http://zebu.uoregon.edu/~j

Selective antagonism of cJun for cancer therapy

The activator protein-1 (AP-1) family of transcription factors modulate a diverse range of cellular signalling pathways into outputs which can be oncogenic or anti-oncogenic. The transcription of relevant genes is controlled by the cellular context, and in particular by the dimeric composition of AP-1. Here, we describe the evidence linking cJun in particular to a range of cancers. This includes correlative studies of protein levels in patient tumour samples and mechanistic understanding of the role of cJun in cancer cell models. This develops an understanding of cJun as a focal point of cancer-altered signalling which has the potential for therapeutic antagonism. Significant work has produced a range of small molecules and peptides which have been summarised here and categorised according to the binding surface they target within the cJun-DNA complex. We highlight the importance of selectively targeting a single AP-1 family member to antagonise known oncogenic function and avoid antagonism of anti-oncogenic function

Mechanical properties and tribological behaviour of electroless Ni–P–Cu coatings on corrosion-resistant alloys under ultrahigh contact stress with sprayed nanoparticles.

Threaded components manufactured from corrosion resistant alloys (CRA's) are vulnerable to galling. This paper develops a test matrix to systematically investigate the mechanical properties and tribological performance of electroless nickel phosphorous coatings on CRA's when subjected to high contact stress. Samples manufactured from 28Cr stainless steel were shot-peened for various periods prior to being electo/electroless coated. The coefficient of friction (CoF) of different coating systems was evaluated via sliding cross-pin method. Various wet and dry lubricants were utilised to examine tribological performance, furthermore the adhesion strength of the coatings was investigated by a bond and pull-off method. The study has shown a significant reduction in CoF for electroless nickel phosphorous coatings with prior shot-peening treatment and sprayed nanoparticles.N/

Have Anglo-Catholics lost their vision for mission agencies? : an empirical enquiry among newly ordained clergy in Britain

Attitude toward Christian mission agencies was investigated in a sample of 827 Anglican clergy ordained in the UK from 2002 to 2006. The Scale of Attitude Toward Mission Agencies (SATMA) consisted of six items related to the work that agencies do, and whether clergy wished to engage with this work. It had a high internal consistency reliability (Cronbach's alpha = 0.80). After controlling for theological liberalism or conservatism, attitudes were most positive among evangelicals and least positive among Anglo-Catholics. Both liberal and conservative Anglo-Catholic clergy showed less positive attitudes toward mission agencies than did other clergy

Structural and functional analysis of the tandem β-zipper interaction of a streptococcal protein with human fibronectin

Bacterial fibronectin-binding proteins (FnBPs) contain a large intrinsically disordered region (IDR) that mediates adhesion of bacteria to host tissues, and invasion of host cells, through binding to fibronectin (Fn). These FnBP IDRs consist of Fn-binding repeats (FnBRs) that form a highly extended tandem β-zipper interaction on binding to the N-terminal domain of Fn. Several FnBR residues are highly conserved across bacterial species, and here we investigate their contribution to the interaction. Mutation of these residues to alanine in SfbI-5 (a disordered FnBR from the human pathogen Streptococcus pyogenes) reduced binding, but for each residue the change in free energy of binding was <2 kcal/mol. The structure of an SfbI-5 peptide in complex with the second and third F1 modules from Fn confirms that the conserved FnBR residues play equivalent functional roles across bacterial species. Thus, in SfbI-5, the binding energy for the tandem β-zipper interaction with Fn is distributed across the interface rather than concentrated in a small number of "hot spot" residues that are frequently observed in the interactions of folded proteins. We propose that this might be a common feature of the interactions of IDRs and is likely to pose a challenge for the development of small molecule inhibitors of FnBP-mediated adhesion to and invasion of host cells