Association between Metabolic Associated Fatty Liver Disease (MAFLD) with treated chronic hepatitis B prognosis

Contexte : un quart des patients infectés par l’hépatite B a un stéatopathie associée et le pronostic de ces patients est mal connu. Par définition le diagnostic de la stéatopathie n’était pas possible en association avec une hépatite B. Ce n’est plus le cas avec la Metabolic associeted fatty liver disease. Objectif : l’objectif de cette étude était d’évaluer l’effet de la MAFLD sur la mortalité toutes causes, la mortalité hépatique et la morbidité hépatique, cardiovasculaire et néoplasique chez les sujets traités pour une infection chronique par le VHB dans la cohorte ANRS CO22 HEPATHER entre 2012 et 2021. Méthode : la population d’étude était constituée des sujets traités à l’inclusion dans la cohorte prospective ANRS CO22 HEPATHER non transplantés hépatiques, non infectés par le VHD et dont le statut vis à vis de la MAFLD pouvait être déterminé. Le FLI était utilisé pour poser le diagnostic de stéatose hépatique. Il a été calculé après imputation multiples des valeurs manquantes de GGT et de triglycérides. Le statut MAFLD était déterminé en utilisant les critères proposés par Eslam et al. Le critère de jugement principal était la survenue du décès toutes causes. Les critères de jugement secondaires étaient la survenue d’une première complication hépatique (dont le CHC), d’une première complication cardiovasculaire ischémique et d’un premier cancer extra-hépatique. Des modèles de Cox ont été réalisés permettant d’ajuster sur les facteurs de confusion identifiés. Résultats : 2180 sujets étaient suivis pendant 7 ans en moyenne. En analyse multivariable, la MAFLD était significativement associée à la mortalité toutes causes (HR = 1,77 [1,24 ; 2,53]). La MAFLD n’était pas significativement associée à la mortalité hépatique (HR = 1,25 [0,70 ; 2,25]) ni à la survenue d’une complication hépatique (HR = 1,32 [0,69 ; 2,52]), d’une complication cardiovasculaire ischémique (HR = 0,63 [0,17 ; 2,25]) ou encore d’un cancer extra-hépatique (HR = 1,42 [0,65 ; 3,13]). Conclusion : dans cette étude de cohorte prospective française de grande taille, la MAFLD était significativement associée à la mortalité chez des sujets traités pour une hépatite virale B chronique.Background: a quarter of patients infected with hepatitis B have associated fatty liver disease and the prognosis of these patients is poorly understood. By definition the diagnosis of fatty liver disease was not possible with hepatitis B. This is no longer the case with Metabolic associeted fatty liver disease. Objective: this study objective was to assess the effect of MAFLD on all-cause mortality, hepatic mortality and hepatic, cardiovascular and neoplastic morbidity in subjects treated for chronic HBV infection in the ANRS CO22 HEPATHER cohort between 2012 and 2021. Methods: the study population consisted of subjects treated at inclusion in the ANRS CO22 HEPATHER prospective cohort who had no history of liver transplantation or co-infection with hepatitis D virus and whose MAFLD status could be determined. The Fatty Liver Index (FLI) was used to diagnose fatty liver disease (FLI> 60). It was calculated after multiple imputation for missing values of GGT and triglycerides. MAFLD status was determined using the criteria proposed by Eslam et al. in 2020. The primary outcome was the occurrence of death from any cause. The secondary outcomes were the occurrence of a first hepatic complication (including HCC), a first ischemic cardiovascular complication and a first extrahepatic cancer. Cox models were performed to adjust for confounding factors. Results: 2180 subjects were followed for 7 years on average. In multivariate analysis adjusted for age, sex and tobacco, MAFLD was significantly associated with all-cause mortality (HR = 1,77 [1,24 ; 2,53]). MAFLD was not associated with hepatic mortality (HR = 1.25 [0.70; 2.25]), hepatic complication (HR = 1.32 [0.69; 2, 52]), ischemic cardiovascular complication (HR = 0.63 [0.17; 2.25]) or extrahepatic cancer (HR = 1.42 [0.65; 3.13] ). Conclusion: in this large French prospective cohort study, MAFLD was significantly associated with mortality in subjects treated for chronic viral hepatitis B

Etudes des microplastiques appliquées à la sécurité sanitaire des produits de la pêche

International audienceLes plastiques ainsi que leurs produits de dégradation comme les microplastiques (MPs) sont présents en quantités importantes dans les Océans. Si les conséquences des MPs sur l’environnement et le biote sont de plus en plus documentées, en revanche peu d’études concernent les organismes aquatiques en tant qu’aliment. Nos recherches portent sur l’impact des MPs sur la sécurité sanitaire des produits de la pêche.Afin de pouvoir réaliser des études comparables, des modes opératoires uniformisés sont nécessaires. Ainsi lors d’une première étude, nous avons cherché à sélectionner parmi un ensemble de protocoles issus de la littérature, celui permettant d’extraire puis d’identifier un maximum de polymères plastiques. La méthode retenue utilise une solution d’hydroxyde de potassium 10% pendant 24H à 60°C. Elle a permis de digérer des échantillons de produits de la mer tout en s’assurant que 14 des 15 types de polymères testés ne sont pas dégradés par la méthode.Parallèlement à cette partie récupération des MPs, des travaux ont été menés en terme d’identification des polymères plastiques par une approche de pyrolyse couplée à un chromatographe en phase gaz et un spectromètre de masse (Py-GC/MS). Cette méthode a fait l’objet d’une validation : estimation de la limite de détection, tests de répétabilité et de reproductibilité. Cette technique pourrait être appliquée en complément d’autres techniques d’identification comme la microspectrométrie Raman ou le FT-IR pour identifier des particules de petite taille comme les fibres ou encore les “pigments”.Par ailleurs, la Py-GC/MS sous réserve d’appliquer des températures de pyrolyse faibles (<100°C) serait également capable de détecter la présence d’additifs dans les polymères. Dans le cadre de cette thématique, un modèle digestif in vitro a été développé pour le poisson et l’Homme. L’application de cet outil à des polymères contenant différents additifs permettra d’évaluer la possibilité d’un relargage de produits chimiques dans le chyme

Occurrence and effects of plastic additives on marine environments and organisms: a review

International audiencePlastics debris, especially microplastics, have been found worldwide in all marine compartments. Much research has been carried out on adsorbed pollutants on plastic pieces and hydrophobic organic compounds (HOC) associated with microplastics. However, only a few studies have focused on plastic additives. These chemicals are incorporated into plastics from which they can leach out as most of them are not chemically bound. As a consequence of plastic accumulation and fragmentation in oceans, plastic additives could represent an increasing ecotoxicological risk for marine organisms. The present work reviewed the main class of plastic additives identified in the literature, their occurrence in the marineenvironment, as well as their effects on and transfers to marine organisms. This work identified polybrominated diphenyl ethers (PBDE), phthalates, nonylphenols (NP), bisphenol A (BPA) and antioxidants as the most common plastic additives found in marine environments. Moreover, transfer of these plastic additives to marine organisms has been demonstrated both in laboratory and field studies. Upcoming research focusing on the toxicity of microplastics should include these plastic additives as potential hazards for marine organisms, and a greater focus on the transport and fate of plastic additives is now required considering that these chemicals may easily leach out from plastics

Development of a Py-GC/MS method and its application to identify marine microplastics

International audiencePlastics are found to be major debris composing marine litter; microplastics (MP, < 5 mm) are found in all marine compartments. The amount of MPs tends to increase with decreasing size leading to a potential misidentification when only visual identification is performed. These last years, pyrolysis coupled with gas chromatography/mass spectrometry (Py-GC/MS) has been used to get information on the composition of polymers with some applications on MP identification. The purpose of this work was to optimize and then validate a Py-GC/MS method, determine limit of detection (LOD) for eight common polymers, and apply this method on environmental MP. Optimization on multiple GC parameters was carried out using polyethylene (PE) and polystyrene (PS) microspheres. The optimized Py-GC/MS method require a pyrolysis temperature of 700 degrees C, a split ratio of 5 and 300 degrees C as injector temperature. Performance assessment was accomplished by performing repeatability and intermediate precision tests and calculating limit of detection (LOD) for common polymers. LODs were all below 1 mug. For performance assessment, identification remains accurate despite a decrease in signal over time. A comparison between identifications performed with Raman micro spectroscopy and with Py-GC/MS was assessed. Finally, the optimized method was applied to environmental samples, including plastics isolated from sea water surface, beach sediments, and organisms collected in the marine environment. The present method is complementary to mu-Raman spectroscopy as Py-GC/MS identified pigment containing particles as plastic. Moreover, some fibers and all particles from sediment and sea surface were identified as plastic

Study of the leaching of additive from microplastics using an in vitro enzymatic digestion model

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Py-GC/MS method development and application for the identification of microplastics from seafood

International audiencePlastics are found to be major debris composing marine litter; microplastics (MP, < 5 mm) are found in all marine compartments. The amount of MPs tends to increase with decreasing size leading to a potential misidentification when only visual identification is performed. These last years, pyrolysis coupled with gas chromatography/mass spectrometry (Py-GC/MS) has been used to get information on the composition of polymers with some applications on MP identification. The purpose of this work was to optimize and then validate a Py-GC/MS method, determine limit of detection (LOD) for eight common polymers, and apply this method on environmental MP. Optimization on multiple GC parameters was carried out using polyethylene (PE) and polystyrene (PS) microspheres. The optimized Py-GC/MS method require a pyrolysis temperature of 700 degrees C, a split ratio of 5 and 300 degrees C as injector temperature. Performance assessment was accomplished by performing repeatability and intermediate precision tests and calculating limit of detection (LOD) for common polymers. LODs were all below 1 mug. For performance assessment, identification remains accurate despite a decrease in signal over time. A comparison between identifications performed with Raman micro spectroscopy and with Py-GC/MS was assessed. Finally, the optimized method was applied to environmental samples, including plastics isolated from sea water surface, beach sediments, and organisms collected in the marine environment. The present method is complementary to mu-Raman spectroscopy as Py-GC/MS identified pigment containing particles as plastic. Moreover, some fibers and all particles from sediment and sea surface were identified as plastic. Graphical abstract

Methodological barriers for extraction and characterization of microplastics in biological matrix

International audiencePollution of the oceans by microplastics, defined as plastic particles of size below < 5mm by the NOAA, represent a major environmental problem worldwide. From an ecotoxicological point of view, the ingestion of microplastics by a wide range of marine organisms leading to substantial impacts on major physiological functions has been shown in several marine vertebrates and invertebrates. To date, only a few studies have investigated the levels of contamination of marine organisms collected in situ, partly due to technical difficulties in isolation and characterization of microplastics in biological samples. The crucial step is the identification of solvent(s) or chemical(s) that efficiently dissolve organic matter and biological tissues without degrading plastic polymers, and this in a time and cost effective way. Most published papers, as well as OSPAR recommendations for the development of a common monitoring protocol for plastic particles in fish and shellfish at the European level, used protocols containing nitric acid to digest the biological tissues, despite reports of substantial polyamide (also known as nylon) degradation with this solvent. In addition, testing a wide range of plastic polymers, and especially those commonly found in the marine environment (the “Big six”), is essential to proposevalidate a common protocol and avoid an underestimation of plastic contents in marine organisms after tissue digestion. In the present study, we reviewed existing approaches and we compared (i) their efficiency in digesting biological matrixes and (ii) their effects on 5 different plastic polymers. Effects on plastics were evaluated through visual inspection, weighing, Pyr-GC/MS and Raman analyses before and after digestion, while tissue digestates were filtered on 1µm-mesh fiber glass filters and observed using a binocular microscope. More research is currently ongoing in our laboratory on a wider range of plastic types (n=15 in total). The aim is to identify and proposevalidate a unique and standardized protocol that could be implemented at the international level to insure relevance and comparison of environmental studies on this topic

Optimization, performance, and application of a pyrolysis-GC/MS method for the identification of microplastics

Plastics are found to be major debris composing marine litter; microplastics (MP, < 5 mm) are found in all marine compartments. The amount of MPs tends to increase with decreasing size leading to a potential misidentification when only visual identification is performed. These last years, pyrolysis coupled with gas chromatography/mass spectrometry (Py-GC/MS) has been used to get information on the composition of polymers with some applications on MP identification. The purpose of this work was to optimize and then validate a Py-GC/MS method, determine limit of detection (LOD) for eight common polymers, and apply this method on environmental MP. Optimization on multiple GC parameters was carried out using polyethylene (PE) and polystyrene (PS) microspheres. The optimized Py-GC/MS method require a pyrolysis temperature of 700 °C, a split ratio of 5 and 300 °C as injector temperature. Performance assessment was accomplished by performing repeatability and intermediate precision tests and calculating limit of detection (LOD) for common polymers. LODs were all below 1 μg. For performance assessment, identification remains accurate despite a decrease in signal over time. A comparison between identifications performed with Raman micro spectroscopy and with Py-GC/MS was assessed. Finally, the optimized method was applied to environmental samples, including plastics isolated from sea water surface, beach sediments, and organisms collected in the marine environment. The present method is complementary to μ-Raman spectroscopy as Py-GC/MS identified pigment containing particles as plastic. Moreover, some fibers and all particles from sediment and sea surface were identified as plastic

Identification of an optimized protocol for extraction and characterization of microplastics in seafood products

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Les microplastiques dans les produits de la mer

International audienceLa pollution des océans par les microplastiques (<5mm) est une préoccupation à l’échelle mondiale, puisque que l’on peut les retrouver dans l’ensemble des compartiments de l’écosystème marin dont le biota, et notamment chez plusieurs vertébrés et invertébrés marins. D'un point de vue écotoxicologique, l'ingestion de microplastiques par un large éventail d'organismes marins peut entraîner des répercussions importantes sur leurs fonctions physiologiques. Plus précisément, en ce qui concerne les produits de la mer destinés à la consommation humaine, les microplastiques peuvent être considérés comme des contaminants. Cependant à ce jour très peu d'études ont été menées sur l'exposition potentielle aux microplastiques par la consommation de produits de la mer. Une première étape importante consiste à utiliser une méthode de récupération qui permet de digérer efficacement différents types de matrices biologiques tout en préservant l'intégrité des polymères pour permettre une identification fiable, rapide, peu onéreuse et efficace. L’impact de six protocoles existants, utilisant des solutions acides, basiques, enzymatiques ou oxydantes pour la digestion de la matière organique, a été étudié sur 15 polymères plastiques. Les plastiques choisis comptent parmi ceux qui sont les plus produits : le polyéthylène (PE), le polypropylène (PP), le chlorure de polyvinyle (PVC), le polyuréthane (PUR), le polystyrène (PS) et du polyamide (PA). Les effets de ces méthodes d’extraction sur les matières plastiques ont été évalués par microscopie, pesée, analyse en Pyrolyse et chromatographie en phase gazeuse couplée à un spectromètre de masse (Pyr-GC/MS) et micro-spectrométrie Raman avant et après la « digestion ». Le protocole le plus prometteur, basé sur l’hydroxyde de potassium, a été sélectionné pour son innocuité sur 14 polymères testés et son efficacité de digestion sur plusieurs matrices biologiques. La méthode sélectionnée a permis d’engager une étude de prévalence des microplastiques dans des produits de la mer fréquemment consommés. Quatre espèces de poissons (sole, maquereau, dorade et cabillaud), et deux espèces de coquillages (moules et coques) ont été prélevées en manche et en atlantique afin de déterminer le niveau de contamination des organismes et d’évaluer le risque d’exposition du consommateur