Characterisation of the genetic diversity of Brucella by multilocus sequencing

<p>Abstract</p> <p>Background</p> <p><it>Brucella </it>species include economically important zoonotic pathogens that can infect a wide range of animals. There are currently six classically recognised species of <it>Brucella </it>although, as yet unnamed, isolates from various marine mammal species have been reported. In order to investigate genetic relationships within the group and identify potential diagnostic markers we have sequenced multiple genetic loci from a large sample of <it>Brucella </it>isolates representing the known diversity of the genus.</p> <p>Results</p> <p>Nine discrete genomic loci corresponding to 4,396 bp of sequence were examined from 160 <it>Brucella </it>isolates. By assigning each distinct allele at a locus an arbitrary numerical designation the population was found to represent 27 distinct sequence types (STs). Diversity at each locus ranged from 1.03–2.45% while overall genetic diversity equated to 1.5%. Most loci examined represent housekeeping gene loci and, in all but one case, the ratio of non-synonymous to synonymous change was substantially <1. Analysis of linkage equilibrium between loci indicated a strongly clonal overall population structure. Concatenated sequence data were used to construct an unrooted neighbour-joining tree representing the relationships between STs. This shows that four previously characterized classical <it>Brucella </it>species, <it>B. abortus</it>, <it>B. melitensis</it>, <it>B. ovis </it>and <it>B. neotomae </it>correspond to well-separated clusters. With the exception of biovar 5, <it>B. suis </it>isolates cluster together, although they form a more diverse group than other classical species with a number of distinct STs corresponding to the remaining four biovars. <it>B. canis </it>isolates are located on the same branch very closely related to, but distinguishable from, <it>B. suis </it>biovar 3 and 4 isolates. Marine mammal isolates represent a distinct, though rather weakly supported, cluster within which individual STs display one of three clear host preferences.</p> <p>Conclusion</p> <p>The sequence database provides a powerful dataset for addressing ongoing controversies in <it>Brucella </it>taxonomy and a tool for unambiguously placing atypical, phenotypically discordant or newly emerging <it>Brucella </it>isolates. Furthermore, by using the phylogenetic backbone described here, robust and rationally selected markers for use in diagnostic assay development can be identified.</p

A stepped wedge cluster randomized control trial of dried blood spot testing to improve the uptake of hepatitis C antibody testing within UK prisons

Background: The prevalence of hepatitis C (HCV) is elevated within prison populations, yet diagnosis in prisons remains low. Dried blood spot testing (DBST) is a simple procedure for the detection of HCV antibodies; its impact on testing in the prison context is unknown. Methods: We carried out a stepped-wedge cluster-randomized control trial of DBST for HCV among prisoners within five male prisons and one female prison. Each prison was a separate cluster. The order in which the intervention (training in use of DBST for HCV testing and logistic support) was introduced was randomized across clusters. The outcome measure was the HCV testing rate by prison. Imputation analysis was carried out to account for missing data. Planned and actual intervention times differed in some prisons; data were thus analysed by intention to treat (ITT) and by observed step times. Results: There was insufficient evidence of an effect of the intervention on testing rate using either the ITT intervention time (OR: 0.84; 95% CI: 0.68�1.03; P = 0.088) or using the actual intervention time (OR: 0.86; 95% CI: 0.71�1.06; P = 0.153). This was confirmed by the pooled results of five imputed data sets. Conclusions: DBST as a stand-alone intervention was insufficient to increase HCV diagnosis within the UK prison setting. Factors such as staff training and allocation of staff time for regular clinics are key to improving service delivery. We demonstrate that prisons can conduct rigorous studies of new interventions, but data collection can be problematic. Trial registration: International Standard Randomized Controlled Trial Number Register (ISRCTN number ISRCTN05628482)

The β6/β7 region of the Hsp70 substrate-binding domain mediates heat-shock response and prion propagation.

Hsp70 is a highly conserved chaperone that in addition to providing essential cellular functions and aiding in cell survival following exposure to a variety of stresses is also a key modulator of prion propagation. Hsp70 is composed of a nucleotide-binding domain (NBD) and substrate-binding domain (SBD). The key functions of Hsp70 are tightly regulated through an allosteric communication network that coordinates ATPase activity with substrate-binding activity. How Hsp70 conformational changes relate to functional change that results in heat shock and prion-related phenotypes is poorly understood. Here, we utilised the yeast [PSI +] system, coupled with SBD-targeted mutagenesis, to investigate how allosteric changes within key structural regions of the Hsp70 SBD result in functional changes in the protein that translate to phenotypic defects in prion propagation and ability to grow at elevated temperatures. We find that variants mutated within the β6 and β7 region of the SBD are defective in prion propagation and heat-shock phenotypes, due to conformational changes within the SBD. Structural analysis of the mutants identifies a potential NBD:SBD interface and key residues that may play important roles in signal transduction between domains. As a consequence of disrupting the β6/β7 region and the SBD overall, Hsp70 exhibits a variety of functional changes including dysregulation of ATPase activity, reduction in ability to refold proteins and changes to interaction affinity with specific co-chaperones and protein substrates. Our findings relate specific structural changes in Hsp70 to specific changes in functional properties that underpin important phenotypic changes in vivo. A thorough understanding of the molecular mechanisms of Hsp70 regulation and how specific modifications result in phenotypic change is essential for the development of new drugs targeting Hsp70 for therapeutic purposes

Don’t look back in anger: the rewarding value of a female face is discounted by an angry expression

The modulating effect of emotional expression on the rewarding nature of attractive and nonattractive female faces in heterosexual men was explored in a motivated viewing paradigm. This paradigm, which is an indicator of neural reward, requires the viewer to expend effort to maintain or reduce image-viewing times. Males worked to extend the viewing time for happy and neutral attractive faces but to reduce the viewing time for the attractive angry faces. Attractive angry faces were rated as more aesthetically pleasing than the nonattractive faces; however, the males worked to reduce their viewing time to a level comparable with the nonattractive neutral and happy faces. Therefore, the addition of an angry expression onto an otherwise attractive face renders it unrewarding and aversive to potential mates. Mildly happy expressions on the nonattractive faces did little to improve their attractiveness or reward potential, with males working to reduce viewing time for all nonattractive faces

The Kinematics and Metallicity of the M31 Globular Cluster System

With the ultimate aim of distinguishing between various models describing the formation of galaxy halos (e.g. radial or multi-phase collapse, random mergers), we have completed a spectroscopic study of the globular cluster system of M31. We present the results of deep, intermediate-resolution, fibre-optic spectroscopy of several hundred of the M31 globular clusters using the Wide Field Fibre Optic Spectrograph (WYFFOS) at the William Herschel Telescope in La Palma, Canary Islands. These observations have yielded precise radial velocities (+/-12 km/s) and metallicities (+/-0.26 dex) for over 200 members of the M31 globular cluster population out to a radius of 1.5 degrees from the galaxy center. Many of these clusters have no previous published radial velocity or [Fe/H] estimates, and the remainder typically represent significant improvements over earlier determinations. We present analyses of the spatial, kinematic and metal abundance properties of the M31 globular clusters. We find that the abundance distribution of the cluster system is consistent with a bimodal distribution with peaks at [Fe/H] = -1.4 and -0.5. The metal-rich clusters demonstrate a centrally concentrated spatial distribution with a high rotation amplitude, although this population does not appear significantly flattened and is consistent with a bulge population. The metal-poor clusters tend to be less spatially concentrated and are also found to have a strong rotation signature.Comment: 33 pages, 20 figure

Angiogenesis in Paget's Disease of the Vulva and the Breast: Correlation with Microvessel Density

Our understanding of the pathogenesis of Paget's disease of the vulva and the breast remains limited. Current evidence supports the fact that angiogenesis plays an important role in the pathogenesis of several diseases. Therefore, we sought to define its role, as correlated with microvessel density, in Paget's disease of the vulva and the breast. Microvessels were analysed using anti-von Willebrand factor antibody in 105 cases of Paget's disease of the vulva and the breast comprising 71 cases of Paget's disease of the vulva, including 8 cases with invasive disease, and 34 cases of Paget's disease of the breast. The latter included 12 cases with DCIS, 5 cases with both DCIS and invasive carcinoma, and 6 with carcinoma alone. Eleven cases had no underlying tumour identified. Increased microvessel density was demonstrated in Paget's disease of the breast with DCIS and with carcinoma alone compared to Paget's disease of the breast alone, P < 0.08 and P < 0.013, respectively. There were no significant differences in microvessel density in the vulval cases. Neovascularisation is an important process in the development of Paget's disease of the breast. Other biological and molecular processes are more involved in the pathogenesis of Paget's disease of the vulva

Imagery or meaning? Evidence for a semantic origin of category-specific brain activity in metabolic imaging

Category-specific brain activation distinguishing between semantic word types has imposed challenges on theories of semantic representations and processes. However, existing metabolic imaging data are still ambiguous about whether these category-specific activations reflect processes involved in accessing the semantic representation of the stimuli, or secondary processes such as deliberate mental imagery. Further information about the response characteristics of category-specific activation is still required. Our study for the first time investigated the differential impact of word frequency on functional magnetic resonance imaging (fMRI) responses to action-related words and visually related words, respectively. First, we corroborated previous results showing that action-relatedness modulates neural responses in action-related areas, while word imageability modulates activation in object processing areas. Second, we provide novel results showing that activation negatively correlated with word frequency in the left fusiform gyrus was specific for visually related words, while in the left middle temporal gyrus word frequency effects emerged only for action-related words. Following the dominant view in the literature that effects of word frequency mainly reflect access to lexico-semantic information, we suggest that category-specific brain activation reflects distributed neuronal ensembles, which ground language and concepts in perception-action systems of the human brain. Our approach can be applied to any event-related data using single-stimulus presentation, and allows a detailed characterization of the functional role of category-specific activation patterns

The Constitutive Relations and the Magnetoelectric Effect for Moving Media

In this paper the constitutive relations for moving media with homogeneous and isotropic electric and magnetic properties are presented as the connections between the generalized magnetization-polarization bivector $$ and the electromagnetic field F. Using the decompositions of F and $\mathcal{M}$, it is shown how the polarization vector P(x) and the magnetization vector M(x) depend on E, B and two different velocity vectors, u - the bulk velocity vector of the medium, and v - the velocity vector of the observers who measure E and B fields. These constitutive relations with four-dimensional geometric quantities, which correctly transform under the Lorentz transformations (LT), are compared with Minkowski's constitutive relations with the 3-vectors and several essential differences are pointed out. They are caused by the fact that, contrary to the general opinion, the usual transformations of the 3-vectors $$, $\mathbf{B}$, $\mathbf{P}$, $\mathbf{M}$, etc. are not the LT. The physical explanation is presented for the existence of the magnetoelectric effect in moving media that essentially differs from the traditional one.Comment: 18 pages, In Ref. [10] here, which corresponds to Ref. [18] in the published paper in IJMPB, Z. Oziewicz's published paper is added. arXiv admin note: text overlap with arXiv:1101.329

The propensity of the bacterial rodlin protein RdlB to form amyloid fibrils determines its function in Streptomyces coelicolor.

Streptomyces bacteria form reproductive aerial hyphae that are covered with a pattern of pairwise aligned fibrils called rodlets. The presence of the rodlet layer requires two homologous rodlin proteins, RdlA and RdlB, and the functional amyloid chaplin proteins, ChpA-H. In contrast to the redundancy shared among the eight chaplins, both RdlA and RdlB are indispensable for the establishment of this rodlet structure. By using a comprehensive biophysical approach combined with in vivo characterization we found that RdlB, but not RdlA, readily assembles into amyloid fibrils. The marked difference in amyloid propensity between these highly similar proteins could be largely attributed to a difference in amino acid sequence at just three sites. Further, an engineered RdlA protein in which these three key amino acids were replaced with the corresponding residues from RdlB could compensate for loss of RdlB and restore formation of the surface-exposed amyloid layer in bacteria. Our data reveal that RdlB is a new functional amyloid and provide a biophysical basis for the functional differences between the two rodlin proteins. This study enhances our understanding of how rodlin proteins contribute to formation of an outer fibrillar layer during spore morphogenesis in streptomycetes