Three-dimensional HepaRG spheroids as a liver model to study human genotoxicity in vitro with the single cell gel electrophoresis assay

International audienceMany efforts have been made in the last 30 years to develop more relevant in vitro models to study genotoxic responses of drugs and environmental contaminants. While 2D HepaRG cells are one of the most promising models for liver toxicology, a switch to 3D cultures that integrate both in vivo architecture and cell-cell interactions has occurred to achieve even more predictive models. preliminary studies have indicated that 3D HepaRG cells are suitable for liver toxicity screening. Our study aimed to evaluate the response of HepaRG spheroids exposed to various genotoxic compounds using the single cell gel electrophoresis assay. HepaRG spheroids were used at 10 days after seeding and exposed for 24 and 48 hours to certain selected chemical compounds (methylmethansulfonate (MMS), etoposide, benzo[a]pyrene (B[a]P), cyclophosphamide (CPA), 7,12-dimethylbenz[a]anthracene (DMBA), 2-acetylaminofluorene (2-AAF), 4-nitroquinoline (4-NQO), 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP), 2-amino-3-methylimidazo[4,5-f]quinolone (IQ), acrylamide, and 2-4-diaminotoluene (2,4-DAT)). After treatment, the comet assay was performed on single cell suspensions and cytotoxicity was determined by the ATP assay. Comet formation was observed for all compounds except IQ, etoposide and 2,4-DAT. Treatment of spheroids with rifampicin increased CYP3A4 activity, demonstrating the metabolic capacity of HepaRG spheroids. These data on genotoxicity in 3D HepaRG spheroids are promising, but further experiments are required to prove that this model can improve the predictivity of in vitro models to detect human carcinogens

Hazard characterization of Alternaria toxins to identify data gaps and improve risk assessment for human health

Fungi of the genus Alternaria are ubiquitous plant pathogens and saprophytes which are able to grow under varying temperature and moisture conditions as well as on a large range of substrates. A spectrum of structurally diverse secondary metabolites with toxic potential has been identified, but occurrence and relative proportion of the different metabolites in complex mixtures depend on strain, substrate, and growth conditions. This review compiles the available knowledge on hazard identification and characterization of Alternaria toxins. Alternariol (AOH), its monomethylether AME and the perylene quinones altertoxin I (ATX-I), ATX-II, ATX-III, alterperylenol (ALP), and stemphyltoxin III (STTX-III) showed in vitro genotoxic and mutagenic properties. Of all identified Alternaria toxins, the epoxide-bearing analogs ATX-II, ATX-III, and STTX-III show the highest cytotoxic, genotoxic, and mutagenic potential in vitro. Under hormone-sensitive conditions, AOH and AME act as moderate xenoestrogens, but in silico modeling predicts further Alternaria toxins as potential estrogenic factors. Recent studies indicate also an immunosuppressive role of AOH and ATX-II; however, no data are available for the majority of Alternaria toxins. Overall, hazard characterization of Alternaria toxins focused, so far, primarily on the commercially available dibenzo-α-pyrones AOH and AME and tenuazonic acid (TeA). Limited data sets are available for altersetin (ALS), altenuene (ALT), and tentoxin (TEN). The occurrence and toxicological relevance of perylene quinone-based Alternaria toxins still remain to be fully elucidated. We identified data gaps on hazard identification and characterization crucial to improve risk assessment of Alternaria mycotoxins for consumers and occupationally exposed workers.The European Partnership for the Assessment of Risks from Chemicals has received funding from the European Union’s Horizon Europe research and innovation program under Grant Agreement No 101057014 and has received co-funding of the authors’ institutions. Views and opinions expressed are, however, those of the author(s) only and do not necessarily reflect those of the European Union or the Health and Digital Executive Agency. Neither the European Union nor the granting authority can be held responsible for them.info:eu-repo/semantics/publishedVersio

Marker residues of dyes in farm fish : study of dye metabolism in fish by in vitro and invivo models

Les colorants de la famille des triarylméthanes sont interdits en aquaculture à cause de leur toxicité. Malgré cela, ils sont toujours utilisés en aquaculture du fait de leurs propriétés antiseptiques et antifongiques, de leurs faibles coûts et de leur disponibilité en tant que produits chimiques pour certaines industries. Afin de protéger les consommateurs, la réglementation européenne impose des contrôles stricts des résidus de substances interdites dans les denrées d’origine animale. En France, pour les colorants à activité pharmacologique, ce contrôle est ciblé sur la présence de résidus de vert de malachite, de cristal violet et de vert brillant dans les produits d’aquaculture, incluant leurs résidus marqueurs métabolisés sous forme de leuco-bases. Pour anticiper le futur de la surveillance des colorants face à d’éventuelles nouvelles pratiques d’usages, cette thèse a exploré différentes approches reposant sur l’analyse du métabolome par chromatographie liquide couplée à la spectrométrie de masse à Haute Résolution et des méthodologies in vitro et in vivo. Les résultats de ce projet apportent des données i) sur le comportement et le devenir de colorants dans les poissons et ii) sur leurs effets, en vue de proposer des marqueurs de traitement issus de l’endo-métabolome et/ou de le xéno-métabolome. Nous avons pu observer une perturbation biologique sur la production d’acides biliaires après traitement au vert de malachite et au victoria pure blue bo (VPBO). Par ailleurs, un métabolite direct issu du traitement de truites au VPBO, le dééthyl-leuco-VPBO, a été souligné comme un résidu marqueur approprié dans le muscle et la peau de poisson. Enfin, une méthode analytique de détection étendue à d’autres familles de colorants, et incluant de nouveaux résidus marqueurs de traitement, a été développée par chromatographie liquide couplée à la spectrométrie de masse en tandem.Dyes from the triarylmethane family are prohibited in aquaculture because of their toxicity. Despite this, they are still used in aquaculture due to their antiseptic and antifungal properties, and to their low costs and availability as chemicals in the industry. In order to protect consumers, European regulations impose strict controls on residues of prohibited substances in food of animal origin. In France, for dyes with pharmacological activity, this control is targeted on the presence of residues of malachite green, crystal violet and brilliant green in aquaculture products, including their marker residues in the form of metabolized leuco-bases. To anticipate the future of dye monitoring due to possible new practices in fish farming, this thesis explored different approaches based on the analysis of the fish metabolome by liquid chromatography coupled with High Resolution mass spectrometry and in vitro or in vivo methodologies. The results of this project provide data i) on the behaviour and fate of dyes in fish and ii) on their effects, with a view to proposing markers of treatments extracted from the endometabolome and / or the xenometabolome. We observed a biological disturbance in the production of bile acids after treatments with malachite green and with victoria pure blue bo (VPBO). Moreover, a direct metabolite resulting from the treatment of trout with VPBO, ie. deethyl-leuco-VPBO, has been highlighted as an appropriate marker residue in fish muscle and skin. Finally, an analytical method extended to several other families of dyes has been developed by means of liquid chromatography coupled with tandem mass spectrometry, and including the new marker residues of treatmen

La dimension argumentative des collocations textuelles en corpus électronique spécialisé au domaine du TAL(N)

Traditionally, the lexical association that leads to the collocational phenomenon is thought to be arbitrary. Nevertheless, when applying the semantico-discursive theory of the Possible Argumentative – PA – on Textual Collocations – TC – this association arises as argumentatively motivated and possibly predictable. We state that TC possibly corresponds to an argumentative collocation, an argumentative relation or an arbitrary collocation. This triple assumption has been tested on the specialized electronic corpus TAL(N), with a multidisciplinary methodology adapted to electronic data (Software and Internet Tools). Our hypothesis has been validated by the results we have obtained. Indeed, they highlight the equal distributions of argumentative collocations and argumentative relations within the corpus compared to arbitrary collocations. This reinforces our intuition of reconsidering the arbitrary nature of collocational associations and motivates our proposition for a description model for composing collocational associations in terms of external argumentative blocks.Traditionnellement, l'association lexicale préexistant au phénomène collocationnel est qualifiée d'arbitraire. Or en soumettant les collocations textuelles – CT – à l'analyse sémantico-discursive des possibles argumentatifs – PA –, modèle de description de la signification lexicale, non seulement cette association s'avère bien souvent argumentativement motivée, mais parfois prédictible. Aussi, nous postulons qu'une CT correspond potentiellement soit à une collocation argumentative, soit à une relation argumentative, soit enfin à une collocation arbitraire. Cette triple hypothèse a été testée sur le corpus électronique spécialisé TAL(N), grâce à une méthodologie pluridisciplinaire adaptée au caractère électronique des données (outils logiciels et Internet). Les résultats obtenus valident notre hypothèse par la présence équilibrée de collocations argumentatives et de relations argumentatives, comparée aux collocations arbitraires. Ces résultats nous confortent dans l'idée de reconsidérer le caractère arbitraire du principe d'association collocationnelle et nous permet d'en proposer une description en termes de blocs d'argumentations externes

Principe de composition sémantico-discursif des collocations lexicales : de l'arbitraire au motivé...

National audienceno abstrac

Résidus marqueurs de colorants dans les poissons d’aquaculture: étude du métabolisme des colorants chez le poisson par des modèles in vitro et in vivo

Dyes from the triarylmethane family are prohibited in aquaculture because of their toxicity. Despite this, they are still used in aquaculture due to their antiseptic and antifungal properties, and to their low costs and availability as chemicals in the industry. In order to protect consumers, European regulations impose strict controls on residues of prohibited substances in food of animal origin. In France, for dyes with pharmacological activity, this control is targeted on the presence of residues of malachite green, crystal violet and brilliant green in aquaculture products, including their marker residues in the form of metabolized leuco-bases. To anticipate the future of dye monitoring due to possible new practices in fish farming, this thesis explored different approaches based on the analysis of the fish metabolome by liquid chromatography coupled with High Resolution mass spectrometry and in vitro or in vivo methodologies. The results of this project provide data i) on the behaviour and fate of dyes in fish and ii) on their effects, with a view to proposing markers of treatments extracted from the endometabolome and / or the xenometabolome. We observed a biological disturbance in the production of bile acids after treatments with malachite green and with victoria pure blue bo (VPBO). Moreover, a direct metabolite resulting from the treatment of trout with VPBO, ie. deethyl-leuco-VPBO, has been highlighted as an appropriate marker residue in fish muscle and skin. Finally, an analytical method extended to several other families of dyes has been developed by means of liquid chromatography coupled with tandem mass spectrometry, and including the new marker residues of treatment.Les colorants de la famille des triarylméthanes sont interdits en aquaculture à cause de leur toxicité. Malgré cela, ils sont toujours utilisés en aquaculture du fait de leurs propriétés antiseptiques et antifongiques, de leurs faibles coûts et de leur disponibilité en tant que produits chimiques pour certaines industries. Afin de protéger les consommateurs, la réglementation européenne impose des contrôles stricts des résidus de substances interdites dans les denrées d’origine animale. En France, pour les colorants à activité pharmacologique, ce contrôle est ciblé sur la présence de résidus de vert de malachite, de cristal violet et de vert brillant dans les produits d’aquaculture, incluant leurs résidus marqueurs métabolisés sous forme de leucobases

La dimension argumentative des collocations textuelles en corpus électronique spécialisé au domaine du TAL (N)

NANTES-BU Lettres (441092102) / SudocSudocFranceF

Chem-AAV: Chemically Modified AAV for Gene Therapy

Chem-AAV: Chemically Modified AAV for Gene Therapy. 20th Annual Meeting of the American-Society-of-Gene-and-Cell-Therapy. (ASGCT

Chem-AAV: Chemically Modified AAV for Gene Therapy

Chem-AAV: Chemically Modified AAV for Gene Therapy. 20th Annual Meeting of the American-Society-of-Gene-and-Cell-Therapy. (ASGCT