CC167 How to Use Pastures

Campaign circular 167: How to use Pastures, talks about the growth of desirable grasses and legumes; how to avoid overgrazing and simply make the best pasture

Genome sequencing and analysis reveals possible determinants of Staphylococcus aureus nasal carriage

<p>Abstract</p> <p>Background</p> <p>Nasal carriage of <it>Staphylococcus aureus </it>is a major risk factor in clinical and community settings due to the range of etiologies caused by the organism. We have identified unique immunological and ultrastructural properties associated with nasal carriage isolates denoting a role for bacterial factors in nasal carriage. However, despite extensive molecular level characterizations by several groups suggesting factors necessary for colonization on nasal epithelium, genetic determinants of nasal carriage are unknown. Herein, we have set a genomic foundation for unraveling the bacterial determinants of nasal carriage in <it>S. aureus.</it></p> <p>Results</p> <p>MLST analysis revealed no lineage specific differences between carrier and non-carrier strains suggesting a role for mobile genetic elements. We completely sequenced a model carrier isolate (D30) and a model non-carrier strain (930918-3) to identify differential gene content. Comparison revealed the presence of 84 genes unique to the carrier strain and strongly suggests a role for Type VII secretion systems in nasal carriage. These genes, along with a putative pathogenicity island (SaPIBov) present uniquely in the carrier strains are likely important in affecting carriage. Further, PCR-based genotyping of other clinical isolates for a specific subset of these 84 genes raise the possibility of nasal carriage being caused by multiple gene sets.</p> <p>Conclusion</p> <p>Our data suggest that carriage is likely a heterogeneic phenotypic trait and implies a role for nucleotide level polymorphism in carriage. Complete genome level analyses of multiple carriage strains of <it>S. aureus </it>will be important in clarifying molecular determinants of <it>S. aureus </it>nasal carriage.</p

Gender accommodative versus transformative approaches: a comparative assessment within a post-harvest fish loss reduction intervention

Technical and social constraints limit value chain actors from equitably engaging in and benefiting from capture fisheries in low-income settings. Extension and development programs often focus on the former, which reflects a technocratic orientation of the fisheries sector and uncertainty about effective ways for development programs to engage with gender and other social constraints. This study presents empirical insights that address these challenges to fisheries development. The study took place in fishing camps in the Barotse Floodplain, Zambia to compare two approaches addressing gender constraints within a broader post-harvest fish loss reduction intervention: an accommodative and a transformative approach. The former embodied a more common ‘practical needs’ set of strategies to ensure female participation, while the latter comprised a communication tool embedded in an action research process to build critical consciousness. Results indicate that the use of a transformative approach led to significant changes in gender equal attitudes and women’s empowerment outcomes compared to only using an accommodative approach. Development programs working in fisheries can apply the findings to engage effectively with gender constraints, especially using transformative approaches to help enable women and men to overcome the social and technical barriers that constrain their lives and livelihoods

A Compensatory Mutation Provides Resistance to Disparate HIV Fusion Inhibitor Peptides and Enhances Membrane Fusion

Fusion inhibitors are a class of antiretroviral drugs used to prevent entry of HIV into host cells. Many of the fusion inhibitors being developed, including the drug enfuvirtide, are peptides designed to competitively inhibit the viral fusion protein gp41. With the emergence of drug resistance, there is an increased need for effective and unique alternatives within this class of antivirals. One such alternative is a class of cyclic, cationic, antimicrobial peptides known as θ-defensins, which are produced by many non-human primates and exhibit broad-spectrum antiviral and antibacterial activity. Currently, the θ-defensin analog RC-101 is being developed as a microbicide due to its specific antiviral activity, lack of toxicity to cells and tissues, and safety in animals. Understanding potential RC-101 resistance, and how resistance to other fusion inhibitors affects RC-101 susceptibility, is critical for future development. In previous studies, we identified a mutant, R5-tropic virus that had evolved partial resistance to RC-101 during in vitro selection. Here, we report that a secondary mutation in gp41 was found to restore replicative fitness, membrane fusion, and the rate of viral entry, which were compromised by an initial mutation providing partial RC-101 resistance. Interestingly, we show that RC-101 is effective against two enfuvirtide-resistant mutants, demonstrating the clinical importance of RC-101 as a unique fusion inhibitor. These findings both expand our understanding of HIV drug-resistance to diverse peptide fusion inhibitors and emphasize the significance of compensatory gp41 mutations. © 2013 Wood et al

Filaggrin-stratified transcriptomic analysis of pediatric skin identifies mechanistic pathways in patients with atopic dermatitis

BackgroundAtopic dermatitis (AD; eczema) is characterized by a widespread abnormality in cutaneous barrier function and propensity to inflammation. Filaggrin is a multifunctional protein and plays a key role in skin barrier formation. Loss-of-function mutations in the gene encoding filaggrin (FLG) are a highly significant risk factor for atopic disease, but the molecular mechanisms leading to dermatitis remain unclear.ObjectiveWe sought to interrogate tissue-specific variations in the expressed genome in the skin of children with AD and to investigate underlying pathomechanisms in atopic skin.MethodsWe applied single-molecule direct RNA sequencing to analyze the whole transcriptome using minimal tissue samples. Uninvolved skin biopsy specimens from 26 pediatric patients with AD were compared with site-matched samples from 10 nonatopic teenage control subjects. Cases and control subjects were screened for FLG genotype to stratify the data set.ResultsTwo thousand four hundred thirty differentially expressed genes (false discovery rate, P < .05) were identified, of which 211 were significantly upregulated and 490 downregulated by greater than 2-fold. Gene ontology terms for “extracellular space” and “defense response” were enriched, whereas “lipid metabolic processes” were downregulated. The subset of FLG wild-type cases showed dysregulation of genes involved with lipid metabolism, whereas filaggrin haploinsufficiency affected global gene expression and was characterized by a type 1 interferon–mediated stress response.ConclusionThese analyses demonstrate the importance of extracellular space and lipid metabolism in atopic skin pathology independent of FLG genotype, whereas an aberrant defense response is seen in subjects with FLG mutations. Genotype stratification of the large data set has facilitated functional interpretation and might guide future therapy development

An Inversion Method for Measuring Beta in Large Redshift Surveys

A precision method for determining the value of Beta= Omega_m^{0.6}/b, where b is the galaxy bias parameter, is presented. In contrast to other existing techniques that focus on estimating this quantity by measuring distortions in the redshift space galaxy-galaxy correlation function or power spectrum, this method removes the distortions by reconstructing the real space density field and determining the value of Beta that results in a symmetric signal. To remove the distortions, the method modifies the amplitudes of a Fourier plane-wave expansion of the survey data parameterized by Beta. This technique is not dependent on the small-angle/plane-parallel approximation and can make full use of large redshift survey data. It has been tested using simulations with four different cosmologies and returns the value of Beta to +/- 0.031, over a factor of two improvement over existing techniques.Comment: 16 pages including 6 figures Submitted to The Astrophysical Journa

Single-qubit lasing in the strong-coupling regime

Motivated by recent ``circuit QED'' experiments we study the lasing transition and spectral properties of single-qubit lasers. In the strong coupling, low-temperature regime quantum fluctuations dominate over thermal noise and strongly influence the linewidth of the laser. When the qubit and the resonator are detuned, amplitude and phase fluctuations of the radiation field are coupled, and the phase diffusion model, commonly used to describe conventional lasers, fails. We predict pronounced effects near the lasing transition, with an enhanced linewidth and non-exponential decay of the correlation functions. We cover a wide range of parameters by using two complementary approaches, one based on the Liouville equation in a Fock state basis, covering arbitrarily strong coupling but limited to low photon numbers, the other based on the coherent-state representation, covering large photon numbers but restricted to weak or intermediate coupling.Comment: 11 pages, 11 figure

Phylogenetic relationships among Staphylococcus species and refinement of cluster groups based on multilocus data

Background: Estimates of relationships among Staphylococcus species have been hampered by poor and inconsistent resolution of phylogenies based largely on single gene analyses incorporating only a limited taxon sample. As such, the evolutionary relationships and hierarchical classification schemes among species have not been confidently established. Here, we address these points through analyses of DNA sequence data from multiple loci (16S rRNA gene, dnaJ, rpoB, and tuf gene fragments) using multiple Bayesian and maximum likelihood phylogenetic approaches that incorporate nearly all recognized Staphylococcus taxa. Results: We estimated the phylogeny of fifty-seven Staphylococcus taxa using partitioned-model Bayesian and maximum likelihood analysis, as well as Bayesian gene-tree species-tree methods. Regardless of methodology, we found broad agreement among methods that the current cluster groups require revision, although there was some disagreement among methods in resolution of higher order relationships. Based on our phylogenetic estimates, we propose a refined classification for Staphylococcus with species being classified into 15 cluster groups (based on molecular data) that adhere to six species groups (based on phenotypic properties). Conclusions: Our findings are in general agreement with gene tree-based reports of the staphylococcal phylogeny, although we identify multiple previously unreported relationships among species. Our results support the general importance of such multilocus assessments as a standard in microbial studies to more robustly infer relationships among recognized and newly discovered lineages

RNase-mediated protein footprint sequencing reveals protein-binding sites throughout the human transcriptome

Although numerous approaches have been developed to map RNA-binding sites of individual RNA-binding proteins (RBPs), few methods exist that allow assessment of global RBP–RNA interactions. Here, we describe PIP-seq, a universal, high-throughput, ribonuclease-mediated protein footprint sequencing approach that reveals RNA-protein interaction sites throughout a transcriptome of interest. We apply PIP-seq to the HeLa transcriptome and compare binding sites found using different cross-linkers and ribonucleases. From this analysis, we identify numerous putative RBP-binding motifs, reveal novel insights into co-binding by RBPs, and uncover a significant enrichment for disease-associated polymorphisms within RBP interaction sites