The Skin and Nose Microbiome and Its Association with Filaggrin Gene Mutations in Pediatric Atopic Dermatitis

BACKGROUND: Interactions between the skin barrier, immune system, and microbiome underlie the development of atopic dermatitis (AD). OBJECTIVE: To investigate the skin and nasal microbiome in relation to filaggrin gene (FLG) mutations. METHODS: A cross-sectional study including 77 children with difficult-to-treat AD. The entire encoding region of FLG was screened for mutations using single molecule molecular inversion probes and next-generation sequencing. Bacterial swabs from the anterior nares, lesional and nonlesional skin were analyzed using 16S rRNA sequencing. For skin samples, additional qPCR was performed for Staphylococcus aureus and Staphylococcus epidermidis. RESULTS: The prevalence of patients with a mutation in FLG was 40%, including 10 different mutations. Analyzing bacterial swabs from all three niches showed a significant effect for both niche and FLG mutation status on the overall microbiome composition. Using a subset analysis to test the effect of FLG mutation status per niche separately did not show a significant association to the microbiome. Shannon diversity and S. aureus abundance were significantly affected by the niche, but not by the presence of an FLG mutation. CONCLUSIONS: Our results suggest only a minor role for FLG mutation status on the overall microbiome, which is rather caused by differences in the present genera than by microbe richness and evenness

Numerical investigation of the in-plane seismic performance of unstrengthened and TRM-strengthened rammed earth walls

The large availability of raw earth around the World led to its extensive use as a building material through history. Thus, earthen materials integrate several historical monuments, but their main use was to build living and working environments for billions of people. On the other hand, past earthquakes revealed their inadequate seismic behavior, which is a matter of concern as a significant percentage of earthen buildings are located in regions with medium to high seismic hazard. Nevertheless, their seismic behavior and the development of efficient strengthening solutions are topics that are not yet sufficiently investigated in the literature. In this context, this study investigates numerically the in-plane seismic behavior of a rammed earth component by means of advanced nonlinear finite element modeling, which included performing nonlinear static (pushover) and nonlinear dynamic analyses. Moreover, the strengthening effectiveness of a low-cost textile-reinforced mortar on such component was also evaluated. The strengthening was observed to increase the load and displacement capacities, to preserve the integrity for higher lateral load levels and to postpone failure without adding significant mass to the system. Furthermore, the pushover analysis was shown to predict reliably the capacities of the models with respect to the incremental dynamic analysis.This work was financed by FEDER funds through the Competitively Factors Operational Programme (COMPETE) and by national funds through the Foundation for Science and Technology (FCT) within the scope of projects POCI-01-0145-FEDER-016737 (PTDC/ECM-EST/2777/2014) and POCI-01-0145-FEDER-007633. The support from grant SFRH/BPD/97082/2013 is also acknowledged

Primary Structure and Catalytic Mechanism of the Epoxide Hydrolase from Agrobacterium radiobacter AD1

The epoxide hydrolase gene from Agrobacterium radiobacter AD1, a bacterium that is able to grow on epichlorohydrin as the sole carbon source, was cloned by means of the polymerase chain reaction with two degenerate primers based on the N-terminal and C-terminal sequences of the enzyme. The epoxide hydrolase gene coded for a protein of 294 amino acids with a molecular mass of 34 kDa. An identical epoxide hydrolase gene was cloned from chromosomal DNA of the closely related strain A. radiobacter CFZ11. The recombinant epoxide hydrolase was expressed up to 40% of the total cellular protein content in Escherichia coli BL21(DE3) and the purified enzyme had a kcat of 21 s-1 with epichlorohydrin. Amino acid sequence similarity of the epoxide hydrolase with eukaryotic epoxide hydrolases, haloalkane dehalogenase from Xanthobacter autotrophicus GJ10, and bromoperoxidase A2 from Streptomyces aureofaciens indicated that it belonged to the α/β-hydrolase fold family. This conclusion was supported by secondary structure predictions and analysis of the secondary structure with circular dichroism spectroscopy. The catalytic triad residues of epoxide hydrolase are proposed to be Asp107, His275, and Asp246. Replacement of these residues to Ala/Glu, Arg/Gln, and Ala, respectively, resulted in a dramatic loss of activity for epichlorohydrin. The reaction mechanism of epoxide hydrolase proceeds via a covalently bound ester intermediate, as was shown by single turnover experiments with the His275 → Arg mutant of epoxide hydrolase in which the ester intermediate could be trapped.