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 supertree approach to shorebird phylogeny

BACKGROUND: Order Charadriiformes (shorebirds) is an ideal model group in which to study a wide range of behavioural, ecological and macroevolutionary processes across species. However, comparative studies depend on phylogeny to control for the effects of shared evolutionary history. Although numerous hypotheses have been presented for subsets of the Charadriiformes none to date include all recognised species. Here we use the matrix representation with parsimony method to produce the first fully inclusive supertree of Charadriiformes. We also provide preliminary estimates of ages for all nodes in the tree. RESULTS: Three main lineages are revealed: i) the plovers and allies; ii) the gulls and allies; and iii) the sandpipers and allies. The relative position of these clades is unresolved in the strict consensus tree but a 50% majority-rule consensus tree indicates that the sandpiper clade is sister group to the gulls and allies whilst the plover group is placed at the base of the tree. The overall topology is highly consistent with recent molecular hypotheses of shorebird phylogeny. CONCLUSION: The supertree hypothesis presented herein is (to our knowledge) the only complete phylogenetic hypothesis of all extant shorebirds. Despite concerns over the robustness of supertrees (see Discussion), we believe that it provides a valuable framework for testing numerous evolutionary hypotheses relating to the diversity of behaviour, ecology and life-history of the Charadriiformes

Magnetic microscopy of topologically protected homochiral domain walls in an ultrathin perpendicularly magnetized Co film

Next-generation concepts for solid-state memory devices are based on current-driven domain wall propagation, where the wall velocity governs the device performance. It has been shown that the domain wall velocity and the direction of travel is controlled by the nature of the wall and its chirality. This chirality is attributed to effects emerging from the lack of inversion symmetry at the interface between a ferromagnet and a heavy metal, leading to an interfacial Dzyaloshinskii-Moriya interaction that can control the shape and chirality of the magnetic domain wall. Here we present direct imaging of domain walls in Pt/Co/AlO$_x$ films using Lorentz transmission electron microscopy, demonstrating the presence of homochiral, and thus topologically protected, N\'{e}el walls. Such domain walls are good candidates for dense data storage, bringing the bit size down close to the limit of the domain wall width

Evolutionary diversification of an ancient gene family (rhs) through C-terminal displacement

<p>Abstract</p> <p>Background</p> <p><it>Rhs </it>genes are prominent features of bacterial genomes that have previously been implicated in genomic rearrangements in <it>E. coli</it>. By comparing <it>rhs </it>repertoires across the Enterobacteriaceae, this study provides a robust explanation of <it>rhs </it>diversification and evolution, and a mechanistic model of how <it>rhs </it>diversity is gained and lost.</p> <p>Results</p> <p><it>Rhs </it>genes are ubiquitous and comprise six structurally distinct lineages within the Enterobacteriaceae. There is considerable intergenomic variation in <it>rhs </it>repertoire; for instance, in <it>Salmonella enterica</it>, <it>rhs </it>are restricted to mobile elements, while in <it>Escherichia coli </it>one <it>rhs </it>lineage has diversified through transposition as older lineages have been deleted. Overall, comparative genomics reveals frequent, independent gene gains and losses, as well as occasional lateral gene transfer, in different genera. Furthermore, we demonstrate that Rhs 'core' domains and variable C-termini are evolutionarily decoupled, and propose that <it>rhs </it>diversity is driven by homologous recombination with circular intermediates. Existing C-termini are displaced by laterally acquired alternatives, creating long arrays of dissociated 'tips' that characterize the appearance of <it>rhs </it>loci.</p> <p>Conclusion</p> <p><it>Rhs </it>repertoires are highly dynamic among Enterobacterial genomes, due to repeated gene gains and losses. In contrast, the primary structures of <it>Rhs </it>genes are evolutionarily conserved, indicating that <it>rhs </it>sequence diversity is driven, not by rapid mutation, but by the relatively slow evolution of novel core/tip combinations. Hence, we predict that a large pool of dissociated <it>rhs </it>C-terminal tips exists episomally and these are potentially transmitted across taxonomic boundaries.</p

Robust microorganisms for biofuel and chemical production from municipal solid waste

BACKGROUND: Worldwide 3.4 billion tonnes of municipal solid waste (MSW) will be produced annually by 2050, however, current approaches to MSW management predominantly involve unsustainable practices like landfilling and incineration. The organic fraction of MSW (OMSW) typically comprises ~ 50% lignocellulose-rich material but is underexplored as a biomanufacturing feedstock due to its highly inconsistent and heterogeneous composition. This study sought to overcome the limitations associated with studying MSW-derived feedstocks by using OMSW produced from a realistic and reproducible MSW mixture on a commercial autoclave system. The resulting OMSW fibre was enzymatically hydrolysed and used to screen diverse microorganisms of biotechnological interest to identify robust species capable of fermenting this complex feedstock. RESULTS: The autoclave pre-treated OMSW fibre contained a polysaccharide fraction comprising 38% cellulose and 4% hemicellulose. Enzymatic hydrolysate of OMSW fibre was high in D-glucose (5.5% w/v) and D-xylose (1.8%w/v) but deficient in nitrogen and phosphate. Although relatively low levels of levulinic acid (30 mM) and vanillin (2 mM) were detected and furfural and 5-hydroxymethylfurfural were absent, the hydrolysate contained an abundance of potentially toxic metals (0.6% w/v). Hydrolysate supplemented with 1% yeast extract to alleviate nutrient limitation was used in a substrate-oriented shake-flask screen with eight biotechnologically useful microorganisms (Clostridium saccharoperbutylacetonicum, Escherichia coli, Geobacillus thermoglucosidasius, Pseudomonas putida, Rhodococcus opacus, Saccharomyces cerevisiae, Schizosaccharomyces pombe and Zymomonas mobilis). Each species' growth and productivity were characterised and three species were identified that robustly and efficiently fermented OMSW fibre hydrolysate without significant substrate inhibition: Z. mobilis, S. cerevisiae and R. opacus, respectively produced product to 69%, 70% and 72% of the maximum theoretical fermentation yield and could theoretically produce 136 kg and 139 kg of ethanol and 91 kg of triacylglycerol (TAG) per tonne of OMSW. CONCLUSIONS: Developing an integrated biorefinery around MSW has the potential to significantly alleviate the environmental burden of current waste management practices. Substrate-oriented screening of a representative and reproducible OMSW-derived fibre identified microorganisms intrinsically suited to growth on OMSW hydrolysates. These species are promising candidates for developing an MSW biorefining platform and provide a foundation for future studies aiming to valorise this underexplored feedstock

Energy justice beyond Borders? Exploring the impact of Brexit on Ireland's all-island energy market

Northern Ireland is part of the United Kingdom, which has left the European Union under the terms of Brexit. The Republic of Ireland is an EU Member State that has remained within the EU. The island of Ireland operates an all-island energy market, and the impact of Brexit on these complex transboundary energy arrangements has been largely overlooked. This study analyses and assesses the significance of Brexit for Ireland's all-island energy market so that the complexities underlying these transboundary circumstances, and the Brexit-related factors acting on them, can be better understood. An energy justice framework is employed that emphasises the consideration of potential distributional, recognitional and procedural injustices in this setting, and that assists in drawing out potentially negative impacts of Brexit on the all-island energy market

EchoBASE: an integrated post-genomic database for Escherichia coli

EchoBASE (http://www.ecoli-york.org) is a relational database designed to contain and manipulate information from post-genomic experiments using the model bacterium Escherichia coli K-12. Its aim is to collate information from a wide range of sources to provide clues to the functions of the approximately 1500 gene products that have no confirmed cellular function. The database is built on an enhanced annotation of the updated genome sequence of strain MG1655 and the association of experimental data with the E.coli genes and their products. Experiments that can be held within EchoBASE include proteomics studies, microarray data, protein–protein interaction data, structural data and bioinformatics studies. EchoBASE also contains annotated information on ‘orphan’ enzyme activities from this microbe to aid characterization of the proteins that catalyse these elusive biochemical reactions

Tips for Teachers of Evidence-based Medicine: Making Sense of Decision Analysis Using a Decision Tree

Decision analysis is a tool that clinicians can use to choose an option that maximizes the overall net benefit to a patient. It is an explicit, quantitative, and systematic approach to decision making under conditions of uncertainty. In this article, we present two teaching tips aimed at helping clinical learners understand the use and relevance of decision analysis. The first tip demonstrates the structure of a decision tree. With this tree, a clinician may identify the optimal choice among complicated options by calculating probabilities of events and incorporating patient valuations of possible outcomes. The second tip demonstrates how to address uncertainty regarding the estimates used in a decision tree. We field tested the tips twice with interns and senior residents. Teacher preparatory time was approximately 90 minutes. The field test utilized a board and a calculator. Two handouts were prepared. Learners identified the importance of incorporating values into the decision-making process as well as the role of uncertainty. The educational objectives appeared to be reached. These teaching tips introduce clinical learners to decision analysis in a fashion aimed to illustrate principles of clinical reasoning and how patient values can be actively incorporated into complex decision making