Draft Genome Sequence of the Principal Etiological Agent of Farmer?s Lung Disease, Saccharopolyspora rectivirgula

Saccharopolyspora rectivirgula is the main cause of farmer's lung disease. The development of recombinant antigens to standardize the serodiagnosis of the disease requires knowledge of the S. rectivirgula genome. We sequenced the genome of an environmental strain, S. rectivirgula DSM 43113. A total of 3,221 proteins were found to be encoded in a short 3.9-Mb genome

Characterization of ply mixing rules for non-symmetric forms of fully orthotropic laminates

Stacking sequence listings are presented for fully orthotropic angle-ply laminates, with up to 21 plies, together with rules for mixing these sequences to form laminates containing any number of plies. The mixing rules are demonstrated through an abridged set of sequences, which are characterized in terms of angle- and cross-ply sub-sequence symmetries. The abridged set of sequences is derived from a new definitive list that supersedes previously published listings. Stacking sequences are presented together with dimensionless parameters from which the bending stiffness terms are readily calculated and an assessment of the bending stiffness efficiency made for angle- and cross-ply sub-sequences. Expressions relating the dimensionless parameters to the well-known lamination parameters are also given, together with graphical representations of feasible domains for all sub-sequence symmetries contained in the definitive list. Feasible domains for extensionally isotropic and fully isotropic laminates are also presented as important sub-sets of fully orthotropic laminates. Finally, examples are given for tapered laminates with fully orthotropic properties, derived from compatible sequences in the definite list

Combined Bacteriophage and Antibiotic Treatment Prevents Pseudomonas aeruginosa Infection of Wild Type and cftr- Epithelial Cells.

With the increase of infections due to multidrug resistant bacterial pathogens and the shortage of antimicrobial molecules with novel targets, interest in bacteriophages as a therapeutic option has regained much attraction. Before the launch of future clinical trials, in vitro studies are required to better evaluate the efficacies and potential pitfalls of such therapies. Here we studied in an ex vivo human airway epithelial cell line model the efficacy of phage and ciprofloxacin alone and in combination to treat infection by Pseudomonas aeruginosa. The Calu-3 cell line and the isogenic CFTR knock down cell line (cftr-) infected apically with P. aeruginosa strain PAO1 showed a progressive reduction in transepithelial resistance during 24 h. Administration at 6 h p.i. of single phage, phage cocktails or ciprofloxacin alone prevented epithelial layer destruction at 24 h p.i. Bacterial regrowth, due to phage resistant mutants harboring mutations in LPS synthesis genes, occurred thereafter both in vitro and ex vivo. However, co-administration of two phages combined with ciprofloxacin efficiently prevented PAO1 regrowth and maintained epithelial cell integrity at 72 p.i. The phage/ciprofloxacin treatment did not induce an inflammatory response in the tested cell lines as determined by nanoString <sup>®</sup> gene expression analysis. We conclude that combination of phage and ciprofloxacin efficiently protects wild type and cftr- epithelial cells from infection by P. aeruginosa and emergence of phage resistant mutants without inducing an inflammatory response. Hence, phage-antibiotic combination should be a safe and promising anti-Pseudomonas therapy for future clinical trials potentially including cystic fibrosis patients

Stacking sequences for extensionally isotropic, fully isotropic and quasi-homogeneous orthotropic laminates

Stacking sequence listings are presented for fully uncoupled Extensionally Isotropic (EILs), Fully Isotropic (FILs) and Quasi-Homogeneous Orthotropic (QHOLs) angle-ply Laminates, with up to 21 plies. All are sub-sets of a definitive list of Fully Orthotropic Laminates (FOLs), containing generally non-symmetric stacking sequences that are characterized in terms of angle- and cross-ply sub-sequence symmetries. Dimensionless parameters are given for each stacking sequence, from which the ABD matrix is readily derived. Expressions relating these dimensionless parameters to the well-known lamination parameters are also given, together with graphical representations of the feasible domains for Pi/3 and Pi/4 EILs and angle-ply QHOLs containing two and three ply orientations. The feasible domain for Pi/3 FILs is represented graphically by a single point, whereas the domain for angle-ply QHOLs containing four ply orientations is represented by a single stacking sequence

Phosphoproteome profiles of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea during exponential growth in axenic cultures

This study describes the gel-free phosphoproteomic analysis of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea grown in vitro under nonlimiting conditions. Using a combination of strong cation exchange and IMAC prior to LC-MS, we identified over 1350 phosphopeptides per fungus representing over 800 phosphoproteins. The preferred phosphorylation sites were found on serine (>80%) and threonine (>15%), whereas phosphorylated tyrosine residues were found at less than 1% in A. brassicicola and at a slightly higher ratio in B. cinerea (1.5%). Biological processes represented principally among the phoshoproteins were those involved in response and transduction of stimuli as well as in regulation of cellular and metabolic processes. Most known elements of signal transduction were found in the datasets of both fungi. This study also revealed unexpected phosphorylation sites in histidine kinases, a category overrepresented in filamentous ascomycetes compared to yeast.The data have been deposited to the ProteomeXchange database with identifie

Microorganisms

The indoor microbial community is a mixture of microorganisms resulting from outdoor ecosystems that seed the built environment. However, the biogeography of the indoor microbial community is still inadequately studied. Dust from more than 3000 dwellings across France was analyzed by qPCR using 17 targets: 10 molds, 3 bacteria groups, and 4 mites. Thus, the first spatial description of the main indoor microbial allergens on the French territory, in relation with biogeographical factors influencing the distribution of microorganisms, was realized in this study. Ten microorganisms out of 17 exhibited increasing abundance profiles across the country: Five microorganisms (Dermatophagoïdes pteronyssinus, Dermatophagoïdes spp., Streptomyces spp., Cladosporium sphaerospermum, Epicoccum nigrum) from northeast to southwest, two (Cryptococcus spp., Alternaria alternata) from northwest to southeast, Mycobacteria from east to west, Aspergillus fumigatus from south to north, and Penicillium chrysogenum from south to northeast. These geographical patterns were partly linked to climate and land cover. Multivariate analysis showed that composition of communities seemed to depend on landscapes, with species related to closed and rather cold and humid landscapes (forests, located in the northeast) and others to more open, hot, and dry landscapes (herbaceous and coastal regions, located in the west). This study highlights the importance of geographical location and outdoor factors that shape communities. In order to study the effect of microorganisms on human health (allergic diseases in particular), it is important to identify biogeographic factors that structure microbial communities on large spatial scales and to quantify the exposure with quantitative tools, such as the multi-qPCR approach

Optimization of radial diffusion coefficients for the proton radiation belt during the CRRES era

Proton flux measurements from the Proton Telescope instrument aboard the CRRES satellite are revisited, and used to drive a radial diffusion model of the inner proton belt at 1.1 ≤ L ≤ 1.65. Our model utilises a physics‐based evaluation of the cosmic ray albedo neutron decay (CRAND) source, and coulomb collisional loss is driven by a drift averaged density model combining results from the International Reference Ionosphere, NRLMSIS‐00 atmosphere and Radio Plasma Imager plasmasphere models, parameterised by solar activity and season. We drive our model using time‐averaged data at L = 1.65 to calculate steady state profiles of equatorial phase space density, and optimise our choice of radial diffusion coefficients based on four defining parameters to minimise the difference between model and data. This is first performed for a quiet period when the belt can be assumed to represent steady state. Additionally, we investigate fitting steady state solutions to time averages taken during active periods where the data exhibits limited deviation from steady state, demonstrated by CRRES measurements following the 24th March 1991 storm. We also discuss a way to make the optimisation process more reliable by excluding periods of variability in plasmaspheric density from any time average. Lastly, we compare our resultant diffusion coefficients to those derived via a similar process in previous work, and diffusion coefficients derived for electrons from ground and in situ observations. We find that higher diffusion coefficients are derived compared with previous work, and suggest more work is required to derive proton diffusion coefficients for different geomagnetic activity levels

Metabolic engineering of astaxanthin biosynthesis in maize endosperm and characterization of a prototype high oil hybrid

Maize was genetically engineered for the biosynthesis of the high value carotenoid astaxanthin in the kernel endosperm. Introduction of a β-carotene hydroxylase and a β-carotene ketolase into a white maize genetic background extended the carotenoid pathway to astaxanthin. Simultaneously, phytoene synthase, the controlling enzyme of carotenogenesis, was over-expressed for enhanced carotenoid production and lycopene ε-cyclase was knocked-down to direct more precursors into the β-branch of the extended ketocarotenoid pathway which ends with astaxanthin. This astaxanthin-accumulating transgenic line was crossed into a high oil- maize genotype in order to increase the storage capacity for lipophilic astaxanthin. The high oil astaxanthin hybrid was compared to its astaxanthin producing parent. We report an in depth metabolomic and proteomic analysis which revealed major up- or down- regulation of genes involved in primary metabolism. Specifically, amino acid biosynthesis and the citric acid cycle which compete with the synthesis or utilization of pyruvate and glyceraldehyde 3-phosphate, the precursors for carotenogenesis, were down-regulated. Nevertheless, principal component analysis demonstrated that this compositional change is within the range of the two wild type parents used to generate the high oil producing astaxanthin hybrid