172 research outputs found

    Precision-Cut Liver Slices of Salmo salar as a tool to investigate the oxidative impact of CYP1A-mediated PCB 126 and 3-methylcholanthrene metabolism

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    a b s t r a c t Fish isolated cell systems have long been used to predict in vivo toxicity of man-made chemicals. In present study, we tested the suitability of Precision-Cut Liver Slices (PCLS) as an alternative to these models that allows the evaluation of a global tissue response to toxicants, to investigate oxidative stress response to cytochrome P450 1A (CYP1A) induction in fish liver. PCLS of Salmo salar were exposed for 21 h to increasing doses of 3-methylcholanthrene (3-MC) and Polychlorobiphenyl 126 (PCB 126). 3-MC (25 lM) strongly induced CYP1A transcription. In dose-response analysis (25-100 lM), EROD activity was strongly increased at intermediate 3-MC concentrations. We found the counter-intuitive decline of EROD at the highest 3-MC doses to result from reversible competition with ethoxyresorufin. No increases of H 2 O 2 production, antioxidant enzymes activities or oxidative damage to lipids were found with 3-MC treatments. PCLS subjected to PCB 126 (2-200 nM) showed increased contamination levels and a parallel increased CYP1A mRNA synthesis and EROD activity. H 2 O 2 production tended to increase but no oxidative damage to lipids was found. As antioxidant enzymes activities declined at the highest PCB 126 dose, it is suggested that longer incubation periods could be required to generate oxidative stress in PCLS

    Insights into metazoan evolution from Alvinella pompejana cDNAs.

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    International audienceBACKGROUND: Alvinella pompejana is a representative of Annelids, a key phylum for evo-devo studies that is still poorly studied at the sequence level. A. pompejana inhabits deep-sea hydrothermal vents and is currently known as one of the most thermotolerant Eukaryotes in marine environments, withstanding the largest known chemical and thermal ranges (from 5 to 105°C). This tube-dwelling worm forms dense colonies on the surface of hydrothermal chimneys and can withstand long periods of hypo/anoxia and long phases of exposure to hydrogen sulphides. A. pompejana specifically inhabits chimney walls of hydrothermal vents on the East Pacific Rise. To survive, Alvinella has developed numerous adaptations at the physiological and molecular levels, such as an increase in the thermostability of proteins and protein complexes. It represents an outstanding model organism for studying adaptation to harsh physicochemical conditions and for isolating stable macromolecules resistant to high temperatures. RESULTS: We have constructed four full length enriched cDNA libraries to investigate the biology and evolution of this intriguing animal. Analysis of more than 75,000 high quality reads led to the identification of 15,858 transcripts and 9,221 putative protein sequences. Our annotation reveals a good coverage of most animal pathways and networks with a prevalence of transcripts involved in oxidative stress resistance, detoxification, anti-bacterial defence, and heat shock protection. Alvinella proteins seem to show a slow evolutionary rate and a higher similarity with proteins from Vertebrates compared to proteins from Arthropods or Nematodes. Their composition shows enrichment in positively charged amino acids that might contribute to their thermostability. The gene content of Alvinella reveals that an important pool of genes previously considered to be specific to Deuterostomes were in fact already present in the last common ancestor of the Bilaterian animals, but have been secondarily lost in model invertebrates. This pool is enriched in glycoproteins that play a key role in intercellular communication, hormonal regulation and immunity. CONCLUSIONS: Our study starts to unravel the gene content and sequence evolution of a deep-sea annelid, revealing key features in eukaryote adaptation to extreme environmental conditions and highlighting the proximity of Annelids and Vertebrates

    Kinetic studies of peroxiredoxin 6 from Arenicola marina: Rapid oxidation by 3 hydrogen peroxide and peroxynitrite but lack of reduction by hydrogen sulfide

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    , respectively, at pH 7.4 and 25°C. Reduction of tert-butyl hydroperoxide was slower. 34 The pK a of the peroxidatic thiol of AmPrx6 was determined as 5

    Évaluations par Cartes Conceptuelles à trous et apprentissage par les pairs

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    Cet article décrit un nouveau dispositif d’évaluation des acquis d’apprentissage basé sur des cartes conceptuelles « à trous » (CCàT) permettant également l’apprentissage par les pairs en grands auditoires durant les tests et une correction automatisée par des formulaires QCM.L’intérêt du dispositif est de garantir une évaluation qualitative (à haut niveau taxonomique) des apprentissages tout en facilitant la conception et la correction de ces évaluations par l’enseignant, même pour de grands groupes d’étudiants (>500) et en favorisant la coopération entre étudiants en amont et pendant les évaluations

    Startle disease in Irish wolfhounds associated with a microdeletion in the glycine transporter GlyT2 gene

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    AbstractDefects in glycinergic synaptic transmission in humans, cattle, and rodents result in an exaggerated startle reflex and hypertonia in response to either acoustic or tactile stimuli. Molecular genetic studies have determined that mutations in the genes encoding the postsynaptic glycine receptor (GlyR) α1 and β subunits (GLRA1 and GLRB) and the presynaptic glycine transporter GlyT2 (SLC6A5) are the major cause of these disorders. Here, we report the first genetically confirmed canine cases of startle disease. A litter of seven Irish wolfhounds was identified in which two puppies developed muscle stiffness and tremor in response to handling. Although sequencing of GLRA1 and GLRB did not reveal any pathogenic mutations, analysis of SLC6A5 revealed a homozygous 4.2kb microdeletion encompassing exons 2 and 3 in both affected animals. This results in the loss of part of the large cytoplasmic N-terminus and all subsequent transmembrane domains due to a frameshift. This genetic lesion was confirmed by defining the deletion breakpoint, Southern blotting, and multiplex ligation-dependent probe amplification (MLPA). This analysis enabled the development of a rapid genotyping test that revealed heterozygosity for the deletion in the dam and sire and three other siblings, confirming recessive inheritance. Wider testing of related animals has identified a total of 13 carriers of the SLC6A5 deletion as well as non-carrier animals. These findings will inform future breeding strategies and enable a rational pharmacotherapy of this new canine disorder

    Evaluating the Potential of Legumes to Mitigate N2O Emissions From Permanent Grassland Using Process-Based Models

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    Funding Information: This modeling study was a joint effort of the Models4Pastures project within the framework of FACCE-JPI. Lutz Merbold and Kathrin Fuchs acknowledge funding received for the Swiss contribution to Models4Pastures (FACCE-JPI project, SNSF funded contract: 40FA40_154245/1) and for the Doc. Mobility fellowship (SNSF funded project: P1EZP2_172121). Lutz Merbold further acknowledges the support received for CGIAR Fund Council, Australia (ACIAR), Irish Aid, the European Union, the Netherlands, New Zealand, Switzerland, UK, USAID, and Thailand for funding to the CGIAR Research Program on Climate Change, Agriculture and Food Security (CCAFS) as well as for the CGIAR Research Program on Livestock. The NZ contributors acknowledge funding from the New Zealand Government Ministry of Primary Industries to support the aims of the Livestock Research Group of the Global Research Alliance on Agricultural Greenhouse Gases and from AgResearch's Strategic Science Investment Fund (the Forages for Reduced Nitrate Leaching (FRNL) research program). The UK partners acknowledge funding by DEFRA and the RCUK projects: N-Circle (BB/N013484/1), UGRASS (NE/M016900/1), and GREENHOUSE (NE/K002589/1). R.M. Rees and C.F.E. Topp also received funding from the Scottish Government Strategic Research Programme. Lorenzo Brilli, Camilla Dibari, and Marco Bindi received funding from the Italian Ministry of Agricultural Food and Forestry Policies (MiPAAF). The FR partners acknowledge funding from CN-MIP project funded by the French National Research Agency (ANR-13-JFAC-0001) and from ADEME (no. 12-60-C0023). Open access funding enabled and organized by Projekt DEAL Funding Information: This modeling study was a joint effort of the Models4Pastures project within the framework of FACCE‐JPI. Lutz Merbold and Kathrin Fuchs acknowledge funding received for the Swiss contribution to Models4Pastures (FACCE‐JPI project, SNSF funded contract: 40FA40_154245/1) and for the Doc. Mobility fellowship (SNSF funded project: P1EZP2_172121). Lutz Merbold further acknowledges the support received for CGIAR Fund Council, Australia (ACIAR), Irish Aid, the European Union, the Netherlands, New Zealand, Switzerland, UK, USAID, and Thailand for funding to the CGIAR Research Program on Climate Change, Agriculture and Food Security (CCAFS) as well as for the CGIAR Research Program on Livestock. The NZ contributors acknowledge funding from the New Zealand Government Ministry of Primary Industries to support the aims of the Livestock Research Group of the Global Research Alliance on Agricultural Greenhouse Gases and from AgResearch's Strategic Science Investment Fund (the Forages for Reduced Nitrate Leaching (FRNL) research program). The UK partners acknowledge funding by DEFRA and the RCUK projects: N‐Circle (BB/N013484/1), UGRASS (NE/M016900/1), and GREENHOUSE (NE/K002589/1). R.M. Rees and C.F.E. Topp also received funding from the Scottish Government Strategic Research Programme. Lorenzo Brilli, Camilla Dibari, and Marco Bindi received funding from the Italian Ministry of Agricultural Food and Forestry Policies (MiPAAF). The FR partners acknowledge funding from CN‐MIP project funded by the French National Research Agency (ANR‐13‐JFAC‐0001) and from ADEME (no. 12‐60‐C0023). Open access funding enabled and organized by Projekt DEAL Publisher Copyright: ©2020. The Authors. Open access funding enabled and organized by Projekt DEALPeer reviewedPublisher PD

    New hyperekplexia mutations provide insight into glycine receptor assembly, trafficking, and activation mechanisms

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    Background: Hyperekplexia mutations have provided much information about glycine receptor structure and function. Results: Weidentified and characterized nine new mutations. Dominant mutations resulted in spontaneous activation, whereas recessive mutations precluded surface expression. Conclusion: These data provide insight into glycine receptor activation mechanisms and surface expression determinants. Significance: The results enhance our understanding of hyperekplexia pathology and glycine receptor structure-function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A

    Sources and Sinks of Greenhouse Gases from European Grasslands and Mitigation Options: The ‘GreenGrass’ Project

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    Adapting the management of grasslands may be used to enhance carbon sequestration into soil, but could also increase N2O and CH4 emissions. In support of the European post-Kyoto policy, the European \u27GreenGrass\u27 project (EC FP5, EVK2-CT2001-00105) has three main objectives: i) to reduce the large uncertainties concerning the estimates of CO2, N2O and CH4 fluxes to and from grassland plots under different climatic conditions and assess their global warming potential, ii) to measure net greenhouse gas (GHG) fluxes for different management which reflect potential mitigation options, iii) to construct a model of the controlling processes to quantify the net fluxes and to evaluate mitigation scenarios by up-scaling to a European level

    The use of biogeochemical models to evaluate mitigation of greenhouse gas emissions from managed grasslands

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    Simulation models quantify the impacts on carbon (C) and nitrogen (N) cycling in grasslandsystems caused by changes in management practices. To support agricultural policies, it is however important to contrast the responses of alternative models, which can differ greatly in their treatment of key processes and in their response to management. We applied eight biogeochemical models at five grassland sites (in France, New Zealand, Switzerland, United Kingdom and United States) to compare the sensitivity of modelled C and N fluxes to changes in the density of grazing animals (from 100% to 50% of the original livestock densities), also in combination with decreasing N fertilization levels (reduced to zero from the initial levels). Simulated multi-model median values indicated that input reduction would lead to an increase in the C sink strength (negative net ecosystem C exchange) in intensive grazing systems: −64 ± 74 g C m−2 yr−1 (animal density reduction) and −81 ± 74 g C m−2 yr−1(N and animal density reduction), against the baseline of −30.5 ± 69.5 g C m−2 yr−1 (LSU [livestock units] ≥ 0.76 ha−1 yr−1). Simulations also indicated a strong effect of N fertilizer reduction on N fluxes, e.g. N2O-N emissions decreased from 0.34 ± 0.22 (baseline) to 0.1 ± 0.05 g N m−2 yr−1 (no N fertilization). Simulated decline in grazing intensity had only limited impact on the N balance. The simulated pattern of enteric methane emissions was dominated by high model-to-model variability. The reduction in simulated offtake (animal intake + cut biomass) led to a doubling in net primary production per animal (increased by 11.6 ± 8.1 t C LSU−1 yr−1 across sites). The highest N2O-N intensities (N2O-N/offtake) were simulated at mown and extensively grazed arid sites. We show the possibility of using grassland models to determine sound mitigation practices while quantifying the uncertainties associated with the simulated outputs
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