A stable fish reporter cell line to study estrogen receptor transactivation by environmental (xeno)estrogens.

International audienceCross-species differences between human and fish estrogen receptor (ER) binding by environmental chemicals have been reported. To study ER transactivation in a fish cellular context, we stably co-transfected the PLHC-1 fish hepatoma cell line with a rainbow trout estrogen receptor (rtER) and the luciferase reporter gene driven by an estrogen response element (ERE). This new cell model, called PELN-rtER (for PLHC-1-ERE-Luciferase-Neomycin), responded to 17beta-estradiol (E2) in a both concentration- and temperature-dependent manner, as well as to environmental ER ligands from different chemical classes: natural and synthetic estrogens, zearalenone metabolites, genistein, alkyphenoles and benzophenone derivatives. The comparison with other in vitro models, i.e. human reporter cell lines (HELN-rtER, MELN) and vitellogenin induction in primary cultures of rainbow trout hepatocytes, showed an overall higher sensitivity of the human cells for a majority of ligands, except for benzophenone derivatives which were active at similar or lower concentrations in fish cells, suggesting species-specificity for these substances. Correlation analyses suggest that the fish cell line is closer to the trout hepatocyte than to the human cell context, and could serve as a relevant mechanistic tool to study ER activation in fish hepatic cellular context

Screening Estrogenic Activities of Chemicals or Mixtures In Vivo Using Transgenic (cyp19a1b-GFP) Zebrafish Embryos

The tg(cyp19a1b-GFP) transgenic zebrafish expresses GFP (green fluorescent protein) under the control of the cyp19a1b gene, encoding brain aromatase. This gene has two major characteristics: (i) it is only expressed in radial glial progenitors in the brain of fish and (ii) it is exquisitely sensitive to estrogens. Based on these properties, we demonstrate that natural or synthetic hormones (alone or in binary mixture), including androgens or progestagens, and industrial chemicals induce a concentration-dependent GFP expression in radial glial progenitors. As GFP expression can be quantified by in vivo imaging, this model presents a very powerful tool to screen and characterize compounds potentially acting as estrogen mimics either directly or after metabolization by the zebrafish embryo. This study also shows that radial glial cells that act as stem cells are direct targets for a large panel of endocrine disruptors, calling for more attention regarding the impact of environmental estrogens and/or certain pharmaceuticals on brain development. Altogether these data identify this in vivo bioassay as an interesting alternative to detect estrogen mimics in hazard and risk assessment perspective

GCN2 phosphorylates HIV-1 integrase and decreases HIV-1 replication by limiting viral integration

AbstractGCN2 is a serine/threonine kinase involved in cellular stress response related to amino acid starvation. Previously, we showed that GCN2 interacts with HIV-1 integrase and is activated during HIV-1 infection. Herein, we identified HIV-1 integrase as a previously unknown substrate of GCN2 in vitro with a major site of phosphorylation at residue S255 located in the C-terminal domain of HIV-1 integrase. The underlying mechanism was investigated and it appeared that the integrase active site was required in order for GCN2 to target the integrase residue S255. Moreover, various integrases from other retroviruses (e.g. MLV, ASV) were also recognized as a substrate by GCN2. In cells, HIV-1 lentiviral particles harboring mutation at integrase position 255 were affected in their replication. Preventing phosphorylation resulted in an increase in infectivity that correlated with an increase in viral DNA integration. Infectivity of MLV was also higher in cells knocked-out for GCN2 suggesting a conserved mechanism to control viral replication. Altogether, our data suggest that GCN2 may constitute a general guardian of genome stability by regulating foreign DNA integration and as such be part of the antiviral armamentarium of the cell.</jats:p

Precision requirements for space-based XCO2 data

Author Posting. © American Geophysical Union, 2007. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 112 (2007): D10314, doi:10.1029/2006JD007659.Precision requirements are determined for space-based column-averaged CO2 dry air mole fraction (XCO2) data. These requirements result from an assessment of spatial and temporal gradients in XCO2, the relationship between XCO2 precision and surface CO2 flux uncertainties inferred from inversions of the XCO2 data, and the effects of XCO2 biases on the fidelity of CO2 flux inversions. Observational system simulation experiments and synthesis inversion modeling demonstrate that the Orbiting Carbon Observatory mission design and sampling strategy provide the means to achieve these XCO2 data precision requirements.This work was supported by the Orbiting Carbon Observatory (OCO) project through NASA’s Earth System Science Pathfinder (ESSP) program. SCO and JTR were supported by a NASA IDS grant (NAG5-9462) to JTR

Mixture effects in samples of multiple contaminants – An inter-laboratory study with manifold bioassays

© 2018 Chemicals in the environment occur in mixtures rather than as individual entities. Environmental quality monitoring thus faces the challenge to comprehensively assess a multitude of contaminants and potential adverse effects. Effect-based methods have been suggested as complements to chemical analytical characterisation of complex pollution patterns. The regularly observed discrepancy between chemical and biological assessments of adverse effects due to contaminants in the field may be either due to unidentified contaminants or result from interactions of compounds in mixtures. Here, we present an interlaboratory study where individual compounds and their mixtures were investigated by extensive concentration-effect analysis using 19 different bioassays. The assay panel consisted of 5 whole organism assays measuring apical effects and 14 cell- and organism-based bioassays with more specific effect observations. Twelve organic water pollutants of diverse structure and unique known modes of action were studied individually and as mixtures mirroring exposure scenarios in freshwaters. We compared the observed mixture effects against component-based mixture effect predictions derived from additivity expectations (assumption of non-interaction). Most of the assays detected the mixture response of the active components as predicted even against a background of other inactive contaminants. When none of the mixture components showed any activity by themselves then the mixture also was without effects. The mixture effects observed using apical endpoints fell in the middle of a prediction window defined by the additivity predictions for concentration addition and independent action, reflecting well the diversity of the anticipated modes of action. In one case, an unexpectedly reduced solubility of one of the mixture components led to mixture responses that fell short of the predictions of both additivity mixture models. The majority of the specific cell- and organism-based endpoints produced mixture responses in agreement with the additivity expectation of concentration addition. Exceptionally, expected (additive) mixture response did not occur due to masking effects such as general toxicity from other compounds. Generally, deviations from an additivity expectation could be explained due to experimental factors, specific limitations of the effect endpoint or masking side effects such as cytotoxicity in in vitro assays. The majority of bioassays were able to quantitatively detect the predicted non-interactive, additive combined effect of the specifically bioactive compounds against a background of complex mixture of other chemicals in the sample. This supports the use of a combination of chemical and bioanalytical monitoring tools for the identification of chemicals that drive a specific mixture effect. Furthermore, we demonstrated that a panel of bioassays can provide a diverse profile of effect responses to a complex contaminated sample. This could be extended towards representing mixture adverse outcome pathways. Our findings support the ongoing development of bioanalytical tools for (i) compiling comprehensive effect-based batteries for water quality assessment, (ii) designing tailored surveillance methods to safeguard specific water uses, and (iii) devising strategies for effect-based diagnosis of complex contamination

Self-paced exercise performance in the heat after pre-exercise cold-fluid ingestion

10.4085/1062-6050-46.6.592Journal of Athletic Training466592 - 599JATT

Continuous thermoregulatory responses to a mass participation 89 km ultramarathon road race

This is the author accepted manuscriptAbstract Purpose: To continuously measure body core temperature (Tc) throughout a mass-participation ultramarathon in sub-elite recreational runners to quantify Tc magnitude and the influence of aerobic fitness and body fat. Methods: Twenty-three participants (19 males, 4 females; age 45 ± 9 y; body mass 72.0 ± 9.3 kg; body fat 26 ± 6%; VO2peak 50 ± 6 ml∙kg-1 ·min-1 ) had gastrointestinal temperature measured during an 89 km ultramarathon. Pre-to-post-race changes in body mass, plasma sodium, and fluid and food recall quantified body water balance. Results: In maximal environmental conditions of 26.3°C and 53% humidity, 21 of the 23 participants finished in 10:28 ± 01:10 hh:mm whilst replacing 49 ± 27% of sweat losses, maintaining plasma sodium (140 ± 3 mmol·l-1 ), and dehydrating by 4.1 ± 1.3%. Mean maximum Tc was 39.0 ± 0.5 (range 38.2-40.1°C) with 90% of race duration ≤39.0°C. Mean maximum ∆Tc was 1.9 ± 0.9 (0.9-2.7°C) with 95% of race duration ≤2.0°C. Over 0-45 km, associations between ΔTc and VO2peak (positive) and body fat (negative) were observed. Over 58-89 km, associations between Tc and VO2peak (negative) and body fat (positive) were observed. Conclusions: Modest Tc responses were observed in recreational ultramarathon runners. Runners with higher levels of aerobic fitness and lower levels of body fat demonstrated the greatest changes in Tc during the first half of the race. Conversely, runners with lower levels of aerobic fitness and higher levels of body fat demonstrated the greatest absolute Tc in the final third of the race