Tnfa signaling through Tnfr2 protects skin against oxidative stress-induced inflammation

14 páginas, 8 figuras.-- Sergio Candel ... et al.-- This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.TNFα overexpression has been associated with several chronic inflammatory diseases, including psoriasis, lichen planus, rheumatoid arthritis, and inflammatory bowel disease. Paradoxically, numerous studies have reported new-onset psoriasis and lichen planus following TNFα antagonist therapy. Here, we show that genetic inhibition of Tnfa and Tnfr2 in zebrafish results in the mobilization of neutrophils to the skin. Using combinations of fluorescent reporter transgenes, fluorescence microscopy, and flow cytometry, we identified the local production of dual oxidase 1 (Duox1)-derived H2O2 by Tnfa- and Tnfr2-deficient keratinocytes as a trigger for the activation of the master inflammation transcription factor NF-κB, which then promotes the induction of genes encoding pro-inflammatory molecules. In addition, pharmacological inhibition of Duox1 completely abrogated skin inflammation, placing Duox1-derived H2O2 upstream of this positive feedback inflammatory loop. Strikingly, DUOX1 was drastically induced in the skin lesions of psoriasis and lichen planus patients. These results reveal a crucial role for TNFα/TNFR2 axis in the protection of the skin against DUOX1-mediated oxidative stress and could establish new therapeutic targets for skin inflammatory disordersThis work was supported by the Spanish Ministry of Science and Innovation (grants BIO2011-23400 and CSD2007-00002 to VM, and PhD fellowship to SC, all co-funded with Fondos Europeos de Desarrollo Regional/European Regional Development Funds), the Fundación Séneca-Murcia (grant 04538/GERM/06 to VM and PhD fellowship to RE-P), Fundação para a Ciência e Tecnologia (PhD fellowship to SdO, SFRH/BD/62674/2009), a Medical Research Council Senior Clinical fellowship to SAR (G0701932), and the European 7th Framework Initial Training Network FishForPharma (PhD fellowship to SDT, PITG-GA-2011-289209).Peer reviewe

Application of the dual-luciferase reporter assay to the analysis of promoter activity in Zebrafish embryos

<p>Abstract</p> <p>Background</p> <p>The dual-luciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. Although this strategy has proved very useful, it does not allow the promoter and gene function to be analyzed in the context of the whole organism.</p> <p>Results</p> <p>Here, we present a rapid and sensitive assay based on the classical dual-luciferase reporter technique which can be used as a new tool to characterize the minimum promoter region of a gene as well as the <it>in vivo </it>response of inducible promoters to different stimuli. We illustrate the usefulness of this system for studying both constitutive (telomerase) and inducible (NF-κB-dependent) promoters. The flexibility of this assay is demonstrated by induction of the NF-κB-dependent promoters using simultaneous microinjection of different pathogen-associated molecular patterns as well as with the use of morpholino-gene mediated knockdown.</p> <p>Conclusion</p> <p>This assay has several advantages compared with the classical <it>in vitro </it>(cell lines) and <it>in vivo </it>(transgenic mice) approaches. Among others, the assay allows a rapid and quantitative measurement of the effects of particular genes or drugs in a given promoter in the context of a whole organism and it can also be used in high throughput screening experiments.</p

Production and mechanism of secretion of interleukin-1b from the marine fish gilthead seabream

©2004. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This document is the Accepted version of a Published Work that appeared in final form in Developmental and Comparative Immunology (DCI). To access the final edited and published work see https://doi.org/10.1016/j.dci.2003.08.002Interleukin-1b (IL-1b) is a secretory cytokine lacking a signal peptide, and does not follow the classical endoplasmic reticulum to Golgi pathway of secretion. Its post-translational processing by IL-1b-converting enzyme (ICE) and subsequent release from activated macrophages requires ATP acting on P2X7 receptors. No information is available on the production and release of fish IL-1b, but the IL-1b gene sequences reported to date lack a conserved ICE recognition site. We show for the first time that lipopolysaccharide (LPS)/macrophage-activating factor (MAF)/bacterial DNA (VaDNA)-primed immune cells of fish accumulate intracellular IL-1b as a ~30 kDa polypeptide (proIL-1b). The combination of LPS and VaDNA was found to be synergistic, suggesting that each ligand is recognized by a different pattern recognition receptor (PRR). More importantly, addition of extracellular ATP does not promote IL-1b secretion by immune cells and fails to induce phosphatidylserine (PS) flip. In contrast, fish SAF-1 fibroblasts shed microvesicles containing a 22 kDa IL-1b form within 30 min of activation with ATP. Notably, the post-translational processing of IL-1b by SAF-1 cells is abrogated by a specific ICE inhibitor

Models of human psoriasis : Zebrafish the newly appointed player

Psoriasis is a human chronic, immune disease with severe cutaneous and systemic manifestations. Its prevalence, among the world population, highly varies with ethnicity and geography, but not sex from remarkable low levels in Asia to 2.3% in Spain, or an impressive 11.5% in Norway. The pathogenesis of psoriasis derives from complex genetic and environmental interactions, which creates aberrant crosstalk between keratinocytes and variated immune cell, resulting in open amplified inflammatory and pro-proliferative circuits. Both, innate and adaptive immune systems are known to be involved in the response at the cellular and humoral levels. Nevertheless, the exact molecular mechanisms are still under debate. Therefore, discovering useful therapeutic targets to stretch the molecular gaps in psoriasis pathogenesis and its associated comorbidities is still mandatory. So far, some mutagenic or pharmacological studies in vitro or using comparative vertebrate models have provided critical molecular insights and directed the human research. Although highly feasible in rodents, the versatile physiology, genetic similarity to humans and outstanding molecular toolbox available, suggest that elaborate forward genetic screenings are far easier to be conducted using the zebrafish model. Thus, in this review, we intend to briefly overview psoriasis and revise in a digested fashion the preclinical research models available, emphasizing the zebrafish as a powerful tool in the study of immune effectors on the same, and how it supports the discovering of new therapies that may help in controlling this widespread disease around the globe.publishedVersionUnit Licence Agreemen

Environmental footprint of organic and conventional grapefruit production irrigated with desalinated seawater in Spain

Citrus fruit production is a major food business with global relevance in the agricultural sector. The surface area of citrus irrigated with desalinated seawater in Spain, the leading citrus producer of Europe, has increased dramatically in the last decade. Desalinated seawater has alleviated water scarcity, but is facing environmental and agronomical challenges due to its high energy consumption and high boron content. The latter particularly affects citrus production due to its sensitivity to boron, since additional water treatment may be required to prevent phytotoxicity damage. The objective of this work was twofold: to quantify and compare, for the first time, the life cycle environmental footprint of (i) organic and conventional grapefruit systems irrigated with desalinated seawater, and (ii) two on-farm boron reduction technologies, namely reverse osmosis and ion exchange resins. Life Cycle Assessment has been used to evaluate the grapefruit production systems and the two technologies. The systems compared had similar characteristics (cultivar and planting density), to enable a fair comparative assessment between organic and conventional woody crops. The results show that the organic grapefruit production had better environmental performance than the conventional system in all selected impact categories and both, land and mass, functional units. The comparison of deboronation technologies showed that ion exchange resins caused a much (one order of magnitude) lower environmental footprint than reverse osmosis. Overall, this study shows that the most environmentally friendly grapefruit system irrigated with desalinated seawater was organic production combined with the use of ion exchange resins for deboronation.This research was supported by the projects SEA4CROP (PID2020-118492RA-C22), funded by MCIN/AEI/10.13039/501100011033 (Spain), and Solution4Farming (PCI2021-122031-2A), funded by MCIN/AEI/10.13039/501100011033 (Spain) and the European Union's NextGenerationEU/PRTR, Horizon2020 Research & Innovation Programme, Joint Call of the Cofund ERA-Nets (grant agreements 696356, 771134, 862665 and 696231). Additional financial support was provided by the AGROALNEXT programme supported by MCIN with funding from European Union NextGenerationEU (PRTR-C17.I1) and by Fundación Séneca with funding from Comunidad Autónoma Región de Murcia (CARM) and the European program NextGenerationEU by the Science and Innovation Missions 2021, Recovery, Transformation and Resilience Plan, NextGenerationEU under the CDTI project SOS-AGUA-XXI" (MIG-20211026). B. Gallego-Elvira acknowledges the support from the Spanish Ministry of Universities (‘Beatriz Galindo’ Fellowship BEAGAL18/00081). Imbernón-Mulero acknowledges the financial support for his PhD work from the project SEA4CROP and the predoctoral program of the Technical University of Cartagena (RV- 484/21, UPCT, Spain)

Multidisciplinary assessment of the agricultural supply of desalinated seawater in south-eastern Spain

Desalinated seawater (DSW) has provided a steady supply of agricultural water for the last decade in south-eastern (SE) Spain, overcoming climatological and hydrological constraints. This article analyses the impacts of the progressive replacement of traditional irrigation water resources with DSW on the main crops of SE Spain, from agronomic, environmental, and economic perspectives, for the first time. The regional magnitude and spatial variability of these impacts have also been evaluated. To that end, six impact indicators were identified and calculated for three water supply scenarios using increasing proportions of DSW, which is representative of the current and possible future situations. The results reflect the high variability of the impact indicators for the different crops. The agronomic impact is favourable but the benefits of reducing water salinity become saturated when DSW exceeds 50 %. The detrimental environmental impact is due to the higher specific energy consumption associated with increased DSW supply, although this is not an intrinsic problem of seawater desalination since it could be mitigated using renewable energies. The economic impact is very sensitive to the cost of DSW and offers heterogeneous results for the different crops. The results show that the combined use of DSW with traditional resources is the most efficient option from the different perspectives of the study, rather than irrigating with DSW alone. The insights from this study could be useful for the assessment of the feasibility of integrating DSW to support agriculture in other regions where desalination plants are being considered to support the water supply.This work is a result of a internship funded by the Autonomous Community of the Region of Murcia through the Fundación Séneca - Agencia de Ciencia y Tecnología de la Región de Murcia (Seneca Foundation - Agency for Science and Technology in the Region of Murcia) and European programme NextGenerationEU. The research was also funded by MCIN/AEI/10.13039/501100011033, project SEA4CROP (PID2020-118492RA-C22). Imbernón-Mulero acknowledges the financial support for his PhD work from the project SEA4CROP and the predoctoral programme of the Technical University of Cartagena (RV-484/21, UPCT, Spain). B. Gallego-Elvira acknowledges the support from the Spanish Ministry of Universities (‘Beatriz Galindo’ Fellowship BEAGAL18/00081). The financial support of Catedra Trasvase y Sostenibilidad – Jose Manuel Claver Valderas and the Autonomous Community of Murcia Region is also acknowledged

Agronomic analysis of the replacement of conventional agricultural water supply by desalinated seawater as an adaptive strategy to water scarcity in South-Eastern Spain

Climate change is affecting water resources in south-eastern Spain, and this mainly affects irrigated agriculture. In this context, seawater desalination is an adaptive strategy that has provided increasing water allotments to agriculture for the last decade, to replace decreasing conventional resources. Farmers are concerned about the agronomic effects of this substitution and its economic consequences. This study focuses on the potential agronomic impacts of the progressive replacement of the irrigation water from the Tagus–Segura transfer (TST) with desalinated seawater (DSW) on the main crops of south-eastern Spain. To that end, five main agronomic concerns were selected and analyzed under three water supply scenarios using increasing rates of DSW (0, 50, and 100%). The results indicated that, in addition to other economic or environmental considerations, sufficiently relevant agronomic aspects exist that need to be considered when replacing the TST supply with DSW. This study evidences the risks of phytotoxicity and soil alkalinization, due to the increase in boron concentration and the imbalance between monovalent and divalent cations in the DSW, respectively, and also a slight increase in the cost of fertilizers. In addition, the irrigation water salinity effect on production and total irrigation requirements was negligible, as both water sources present sufficiently low salinity. The detrimental effects were mitigated under a partial replacement scenario, so the blended use of DSW with conventional resources seems the most recommendable option for its agricultural management, rather than irrigating with DSW alone.This work is a result of the AGROALNEXT program and was supported by MCIN with funding from European Union NextGenerationEU (PRTR-C17.I1) and by Fundación Séneca with funding from Comunidad Autónoma Región de Murcia (CARM). The research was also funded by MCIN/AEI/10.13039/501100011033, project SEA4CROP (PID2020-118492RA-C22). Imbernón-Mulero acknowledges support for his Ph.D. work from the project SEA4CROP and the predoctoral program of the Technical University of Cartagena (RV-484/21, UPCT, Spain). B. Gallego-Elvira acknowledges support from the Spanish Ministry of Universities (“Beatriz Galindo” Fellowship BEAGAL18/00081)