Increased fibrosis in a mouse model of anti-laminin 332 mucous membrane pemphigoid remains unaltered by inhibition of aldehyde dehydrogenase

Mucous membrane pemphigoid (MMP) is an autoimmune blistering disease characterized by autoantibodies against the basal membrane zone of skin and surface-close epithelia and predominant mucosal lesions. The oral cavity and conjunctivae are most frequently affected, albeit clinical manifestations can also occur on the skin. MMP-associated lesions outside the oral cavity typically lead to scarring. Mechanisms underlying scarring are largely unknown in MMP and effective treatment options are limited. Herein, we assessed the collagen architecture in tissue samples of an antibody-transfer mouse model of anti-laminin-332 MMP. In MMP mice, increased collagen fibril density was observed in skin and conjunctival lesions compared to mice injected with normal rabbit IgG. The extracellular matrix of MMP skin samples also showed altered post-translational collagen cross-linking with increased levels of both lysine- and hydroxylysine-derived collagen crosslinks supporting the fibrotic phenotype in experimental MMP compared to control animals. In addition, we evaluated a potential anti-fibrotic therapy in experimental anti-laminin-332 MMP using disulfiram, an inhibitor of the aldehyde dehydrogenase (ALDH), which has been implicated in immune-mediated mucosal scarring. In addition, disulfiram also acts as a copper chelator that was shown to block lysyl oxidase activity, an enzyme involved in formation of collagen crosslinks. Topical use of disulfiram (300 μM in 2 [w/v] methocel) did not improve ocular lesions in experimental MMP over the 12-day treatment period in disulfiram-treated mice compared to vehicle-treated mice (n=8/group). Furthermore, C57BL6/J mice (n=8/group) were treated prophylactically with 200 mg/kg p.o. disulfiram or the solvent once daily over a period of 12 days. Systemic treatment did not show any reduction in the severity of oral and ocular lesions in MMP mice, albeit some improvement in skin lesions was observed in disulfiram- vs. vehicle-treated mice (p=0.052). No reduction in fibrosis was seen, as assessed by immunohistochemistry. Whilst blocking of ALDH failed to significantly ameliorate disease activity, our data provide new insight into fibrotic processes highlighting changes in the collagenous matrix and cross-linking patterns in IgG-mediated MMP

Aging and gate bias effects on TID sensitivity of wide bandgap power devices

The effect of oxide stress on the total ionizing dose (TID) radiation sensitivity of silicon carbide (SiC) power MOSFETS and TID sensitivity of gallium nitride (GaN) power transistor is reported. Difference in TID response for stressed and unstressed devices was observed

Deconstructing design thinking as a tool for the implementation of a population health initiative

BACKGROUND: The translation of evidence-based practices and rapid uptake of innovations into global health practice is challenging. Design thinking is a consultative process involving multiple stakeholders and has been identified as a promising solution to create and apply implementation strategies in complex environments like health systems. METHODS: We conducted a process evaluation of a real-world example, namely an initiative to innovate hypertension screening, diagnosis and care in Sao Paulo, Brazil. The parameters of the evaluation were informed by a specification rubric and categorization system, recommended for the investigation of implementation strategies, and the double-diamond conceptual framework to describe and examine the strategic architecture and nature of the design thinking approach, with particular emphasis on identifying potential areas of "value-add" particular to the approach. The retrospective evaluation was performed by an independent partner who had not been involved in the setting up and implementation of the design thinking process. RESULTS: The evaluation unveiled a dense catalogue of strategically driven, mostly theoretically based, activities involving all identified health system stakeholders including patients. Narrative reconstruction illuminated the systematic and coherent nature of this approach, with different resulting actions progressively accounting for all relevant layers of the health system to engineer a broad selection of specific implementation solutions. The relevance of the identified features and the mechanics used to promote more successful implementation practices was manifested in several distinct ways: design thinking offered a clear direction on which innovations really mattered and when, as well as several new dimensions for consideration in the development of an innovation mindset amongst stakeholders. It thereby promoted relationship quality in terms of familiarity and trust, and commitment to evidence-based enquiry and action. Design thinking was also able to navigate the territory between the need for intervention "fidelity" versus "adaptation" and provide the operational know-how to face familiar implementation hurdles. Lastly, it brought a new kind of skill set to the public health stakeholders that incorporated diplomacy, multidisciplinary approaches and management sciences-skills that are considered necessary but not yet widely taught as part of public health training. CONCLUSIONS: Design thinking is a sound and viable tool to use as part of an implementation strategy for engaging with health system stakeholders and successfully translating evidence-based practices and new innovations into routine practice, thereby addressing an important knowledge-practice gap and, more broadly, contributing to the strategic repertoire available to implementation science

Assessing plant diversity and composition in grasslands across spatial scales: the standardised EDGG sampling methodology

This paper presents the details of the EDGG sampling methodology and its underlying rationales. The methodology has been applied during EDGG Research Expeditions and EDGG Field Workshops since 2009, and has been subsequently adopted by various other researchers. The core of the sampling are the EDGG Biodiversity Plots, which are 100‐m2 squares comprising, in two opposite corners, nested‐plot series of 0.0001, 0.001, 0.01, 0.1, 1 and 10 m2 square plots, in which all terricolous vascular plants, bryophytes and lichens are recorded using the shoot presence method. In the 10‐m2 plots, species cover is also estimated as a percentage and various environmental and structural parameters are recorded. Usually the EDGG Biodiversity Plots are complemented by the sampling of additional 10 m2 normal plots with the same parameters as the 10‐m2 corners of the first, allowing coverage of a greater environmental diversity and the achievement of higher statistical power in the subsequent analyses for this important grain size. The EDGG sampling methodology has been refined over the years, while its core has turned out to generate high‐quality, standardised data in an effective manner, which facilitates a multitude of analyses. In this paper we provide the current versions of our guidelines, field forms and data entry spreadsheets, as open‐access Online Resources to facilitate the easy implementation of this methodology by other researchers. We also discuss potential future additions and modifications to the approach, among which the most promising are the use of stratified‐random methods to a priori localise the plots and ideas to sample invertebrate taxa on the same plots and grain sizes, such as grasshoppers (Orthoptera) and vegetation‐dwelling spiders (Araneae). As with any other method, the EDGG sampling methodology is not ideal for every single purpose, but with its continuous improvements and its flexibility, it is a good multi‐ purpose approach. A particularly advantageous element, lacking in most other sampling schemes, including classical phytosociogical sampling, is the multi‐scale and multi‐taxon approach, which provides data that allow for deeper understanding of the generalities and idiosyncrasies of biodiversity patterns and their underlying drivers across scales and taxa

The whole and its parts : why and how to disentangle plant communities and synusiae in vegetation classification

Most plant communities consist of different structural and ecological subsets, ranging from cryptogams to different tree layers. The completeness and approach with which these subsets are sampled have implications for vegetation classification. Non‐vascular plants are often omitted or sometimes treated separately, referring to their assemblages as “synusiae” (e.g. epiphytes on bark, saxicolous species on rocks). The distinction of complete plant communities (phytocoenoses or holocoenoses) from their parts (synusiae or merocoenoses) is crucial to avoid logical problems and inconsistencies of the resulting classification systems. We here describe theoretical differences between the phytocoenosis as a whole and its parts, and outline consequences of this distinction for practise and terminology in vegetation classification. To implement a clearer separation, we call for modifications of the International Code of Phytosociological Nomenclature and the EuroVegChecklist. We believe that these steps will make vegetation classification systems better applicable and raise the recognition of the importance of non‐vascular plants in the vegetation as well as their interplay with vascular plants

Transgenic Expression of the Anti-parasitic Factor TEP1 in the Malaria Mosquito Anopheles gambiae

Mosquitoes genetically engineered to be resistant to Plasmodium parasites represent a promising novel approach in the fight against malaria. The insect immune system itself is a source of anti-parasitic genes potentially exploitable for transgenic designs. The Anopheles gambiae thioester containing protein 1 (TEP1) is a potent anti-parasitic protein. TEP1 is secreted and circulates in the mosquito hemolymph, where its activated cleaved form binds and eliminates malaria parasites. Here we investigated whether TEP1 can be used to create malaria resistant mosquitoes. Using a GFP reporter transgene, we determined that the fat body is the main site of TEP1 expression. We generated transgenic mosquitoes that express TEP1r, a potent refractory allele of TEP1, in the fat body and examined the activity of the transgenic protein in wild-type or TEP1 mutant genetic backgrounds. Transgenic TEP1r rescued loss-of-function mutations, but did not increase parasite resistance in the presence of a wild-type susceptible allele. Consistent with previous reports, TEP1 protein expressed from the transgene in the fat body was taken up by hemocytes upon a challenge with injected bacteria. Furthermore, although maturation of transgenic TEP1 into the cleaved form was impaired in one of the TEP1 mutant lines, it was still sufficient to reduce parasite numbers and induce parasite melanization. We also report here the first use of Transcription Activator Like Effectors (TALEs) in Anopheles gambiae to stimulate expression of endogenous TEP1. We found that artificial elevation of TEP1 expression remains moderate in vivo and that enhancement of endogenous TEP1 expression did not result in increased resistance to Plasmodium. Taken together, our results reveal the difficulty of artificially influencing TEP1-mediated Plasmodium resistance, and contribute to further our understanding of the molecular mechanisms underlying mosquito resistance to Plasmodium parasites

Honey bee foraging distance depends on month and forage type

To investigate the distances at which honey bee foragers collect nectar and pollen, we analysed 5,484 decoded waggle dances made to natural forage sites to determine monthly foraging distance for each forage type. Firstly, we found significantly fewer overall dances made for pollen (16.8 %) than for non-pollen, presumably nectar (83.2 %; P < 2.2 × 10−23). When we analysed distance against month and forage type, there was a significant interaction between the two factors, which demonstrates that in some months, one forage type is collected at farther distances, but this would reverse in other months. Overall, these data suggest that distance, as a proxy for forage availability, is not significantly and consistently driven by need for one type of forage over the other

Database Species-Area Relationships in Palaearctic Grasslands.

The database collects the data resulting from the sampling of species-area relationships (SARs) in grassland communities in the Palaearctic. The core features of the database are the complete data from the EDGG Research Expeditions, but similar data from other studies are also included. Main features of the majority of relevés are: (i) they are part of nested-plot series (typically 0.0001-100 m²), (ii) bryophytes and lichens are treated comprehensively; (iii) detailed soil and other environmental data

Adaptive remodeling of the bacterial proteome by specific ribosomal modification regulates Pseudomonas infection and niche colonisation

Post-transcriptional control of protein abundance is a highly important, underexplored regulatory process by which organisms respond to their environments. Here we describe an important and previously unidentified regulatory pathway involving the ribosomal modification protein RimK, its regulator proteins RimA and RimB, and the widespread bacterial second messenger cyclic-di-GMP (cdG). Disruption of rimK affects motility and surface attachment in pathogenic and commensal Pseudomonas species, with rimK deletion significantly compromising rhizosphere colonisation by the commensal soil bacterium P. fluorescens, and plant infection by the pathogens P. syringae and P. aeruginosa. RimK functions as an ATP-dependent glutamyl ligase, adding glutamate residues to the C-terminus of ribosomal protein RpsF and inducing specific effects on both ribosome protein complement and function. Deletion of rimK in P. fluorescens leads to markedly reduced levels of multiple ribosomal proteins, and also of the key translational regulator Hfq. In turn, reduced Hfq levels induce specific downstream proteomic changes, with significant increases in multiple ABC transporters, stress response proteins and non-ribosomal peptide synthetases seen for both ΔrimK and Δhfq mutants. The activity of RimK is itself controlled by interactions with RimA, RimB and cdG. We propose that control of RimK activity represents a novel regulatory mechanism that dynamically influences interactions between bacteria and their hosts; translating environmental pressures into dynamic ribosomal changes, and consequently to an adaptive remodeling of the bacterial proteome

Does plant diversity affect the water balance of established grassland systems like in manipulative biodiversity experiments?

Land-use intensification and biodiversity loss are known drivers of the water cycle but their interactions are unclear. We investigated how evapotranspiration (ETa), downward water flux (DF), and capillary rise (CR) in topsoil and subsoil are related to land-use and plant diversity in established, commercially managed grassland and compared these results to findings from an experiment in which plant diversity was manipulated. In three Central European regions (“Biodiversity Exploratories”), we studied 29 grassland plots (50 m x 50 m; 9-11 plots per region). Land-use intensity increases in the order, pasture &lt; mown pasture &lt; meadow. In 2010-2015, we measured soil moisture, meteorological conditions, plant species richness, number of species in the functional groups of grasses, herbs, and legumes, and root biomass. ETa, DF, and CR were calculated for two soil layers with a soil water balance model. Land-use and biodiversity effects on water fluxes were analyzed with repeated-measures analysis of variance. Land-use intensity did not affect water fluxes. Species richness did not influence DF and CR. ETa from topsoil decreased with increasing species richness while ETa from subsoil increased. Opposing effects on ETa in the two soil layers were also observed for the number of herbs and legumes. In manipulative biodiversity experiments, such opposing effects were explained by higher biomass in species-rich mixtures, which increases shading of topsoil and reduces evaporation. In subsoil, deeper roots in species-rich mixtures increased transpiration. In the commercially managed grasslands, biomass and species richness correlated negatively because fertilizer application increased biomass and decreased species richness. Thus, similar effects of biodiversity on water fluxes in commercially managed and experimentally manipulated grassland had different reasons. We speculate that improved infiltration and enhanced bioturbation in species-rich grassland explained our observations