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Study of intracellular anabolism of 5-fluorouracil and incorporation in nucleic acids based on an LC-HRMS method

Journal of Pharmaceutical Analysis 2021年第1期11卷 77-87页

作者： christelle machon Frederic Catez Nicole Dalla Venezia Floriane Vanhalle Laetitia Guyot Anne Vincent Maxime Garcia Beatrice Roy Jean-Jacques Diaz Jerome Guitton Laboratoire de Chimie Analytique Faculte de Pharmacie de LyonUniversite Lyon 18 Avenue RockefellerLyon69373France Laboratoire de Biochimie et de Pharmaco-toxicologie Centre Hospitalier Lyon-SudeHCLPierre Benite69495France Inserm U1052 CNRS UMR5286 Centre de Recherche en Cancerologie de LyonF-69000 LyonFrance.Centre Leon BerardF-69008 LyonFranceUniversite de Lyon 1F-69000 LyonFrance Institut des Biomolecules Max Mousseron(IBMM) UMR 5247 CNRSUniversite de MontpellierENSCMCampus TrioletMontpellierFrance Laboratoire de toxicologie Faculte de pharmacie de Lyon8 avenue RockefellerF-69373 LyonFrance

5-Fluorouracil(5-FU)is an anticancer drug extensively used for different *** metabolic activation leads to several nucleoside and nucleotide metabolites essential to exert its cytotoxic activity on multiple cellular targets such as enzymes,DNA and *** this paper,we describe the development of a method based on liquid chromatography coupled with high resolution mass spectrometry suitable for the simultaneous determination of the ten anabolic metabolites(nucleoside,nucleotide and sugar nucleotide)of *** chromatographic separationwas optimized on a porous graphitic carbon column allowing the analysis of the metabolites of 5-FU as well as endogenous *** detection was performed on an Orbitrap■tandem mass *** of the method was verified in intracellular content and in RNA *** limit of detection was equal to 12 pg injected on column for nucleoside metabolites of 5-FU and 150 pg injected on column for mono-and tri-phosphate nucleotide *** effect was evaluated in cellular contents,DNA and RNA extracts for nucleoside and nucleotides *** method was successfully applied to i)measure the proportion of each anabolic metabolite of 5-FU in cellular contents,ii)follow the consequence of inhibition of enzymes on the endogenous nucleotide pools,iii)study the incorporation of metabolites of 5-FU into RNA and DNA,and iv)to determine the incorporation rate of 5-FUrd into 18 S and 28 S sub-units of rRNA.

关键词： 5-Fluorouracil LC-MS-HRMS Nucleotide RNA DNA Incorporation rate

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Direct impacts of recent climate warming on insect populations

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Integrative Zoology 2010年第2期5卷 132-142页

作者： christelle ROBINET Alain ROQUES National Institute for Agricultural Research INRA UR633OrléansFrance

Effects of recent climate change have already been detected in many species,and,in particular,in *** present paper reviews the key impacts of global warming on insect development and *** effects of climate change appear to be much more complex than a simple linear response to an average increase in *** can differ between seasons and bioclimatic *** flight periods,enhanced winter survival and acceleration of development rates are the major insect *** response of insects and hosts to warming up might also lead to disruption of their phenological synchrony,but adaptive genetic processes are likely to quickly restore this *** a number of cases,warming results in removing or relocating the barriers that limit present species’*** is also likely to facilitate the establishment and spread of invasive alien ***,knowledge gaps are identified and future research interests are suggested.

关键词： adaptation alien species climate warming phenology range distribution

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NRF2 signaling cascade in amyotrophic lateral sclerosis:bridging the gap between promise and reality

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Neural Regeneration Research 2024年第5期19卷 1006-1012页

作者： Pauline Tarot christelle Lasbleiz Jean-Charles Liévens MMDN Univ MontpellierEPHEINSERMMontpellierFrance CNRS ParisFranceEPHEINSERMMontpellierFrance

Amyotrophic lateral sclerosis is a very disabling disease due to the degeneration of motor *** include muscle weakness and atrophy,spasticity,and progressive ***,there is no treatment to reverse damage to motor neurons and cure amyotrophic lateral *** only two treatments actually approved,riluzole and edaravone,have shown mitigated beneficial *** difficulty to find a cure lies in the complexity and multifaceted pattern of amyotrophic lateral sclerosis *** mechanisms,abnormal RNA metabolism,nucleocytoplasmic transport defects,accumulation of unfolded protein,and mitochondrial dysfunction would in fine induce oxidative damage and vice versa.A potent therapeutic strategy will be to find molecules that break this vicious *** the nuclear factor erythroid-2 related factor 2 signaling may fulfill this objective since nuclear factor erythroid-2 related factor 2 has a multitarget profile controlling antioxidant defense,mitochondrial functioning,and *** here discuss the interest of developing nuclear factor erythroid-2 related factor 2-based therapy in regard to the pathophysiological mechanisms and we provide a general overview of the attempted clinical assays in amyotrophic lateral sclerosis.

关键词： amyotrophic lateral sclerosis C9orf72 NRF2 oxidative defense oxidative stress sulforaphane superoxide dismutase 1 TDP43

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Oxidation state specific analysis of arsenic species in tissues of wild-type and arsenic(+3 oxidation state)methyltransferase-knockout mice

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Journal of Environmental Sciences 2016年第11期28卷 104-112页

作者： Jenna M.Currier christelle Douillet Zuzana Drobná Miroslav Styblo Curriculum in Toxicology University of North Carolina at Chapel Hill Department of Nutrition Gillings School of Global Public Health University of North Carolina at Chapel Hill

Arsenic methyltransferase（As3mt） catalyzes the conversion of inorganic arsenic（i As） to its methylated metabolites, including toxic methylarsonite（MAs~Ⅲ） and dimethylarsinite（DMAs~Ⅲ）. Knockout（KO） of As3 mt was shown to reduce the capacity to methylate i As in mice. However, no data are available on the oxidation states of As species in tissues of these mice. Here, we compare the oxidation states of As species in tissues of male C57BL/6 As3mt-KO and wild-type（WT） mice exposed to arsenite（iA s~Ⅲ） in drinking water. WT mice were exposed to50 mg/L As and As3mt-KO mice that cannot tolerate 50 mg/L As were exposed to 0, 15, 20, 25 or30 mg/L As. iA s~Ⅲaccounted for 53% to 74% of total As in liver, pancreas, adipose, lung, heart, and kidney of As3mt-KO mice; tri- and pentavalent methylated arsenicals did not exceed 10% of total As. Tissues of WT mice retained iA s and methylated arsenicals： iA s~Ⅲ, MAs~Ⅲand DMAs~Ⅲ represented 55%‐68% of the total As in the liver, pancreas, and brain. High levels of methylated species, particularly MAs~Ⅲ, were found in the intestine of WT, but not As3mt-KO mice,suggesting that intestinal bacteria are not a major source of methylated As. Blood of WT mice contained significantly higher levels of As than blood of As3mt-KO mice. This study is the first to determine oxidation states of As species in tissues of As3mt-KO mice. Results will help to design studies using WT and As3mt-KO mice to examine the role of iA s methylation in adverse effects of iA s exposure.

关键词： Arsenic speciation analysis Hydride generation-cryotrapping-atomic absorption spectrometry Arsenic(+ 3 oxidation state) methyltransferase As3mt knockout mice

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Impact of a Phosphate Amendment on the Environmental Availability and Phytoavailability of Cd and Pb in Moderately and Highly Carbonated Kitchen Garden Soils

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Pedosphere 2017年第3期27卷 588-605页

作者： Christophe WATERLOT christelle PRUVOT Franck MAROT Francis DOUAY Yncréa Hauts-de-France Laboratoire Génie Civil et géo Environnement (LGCgE) Agence De l’Environnement et de la Matrise de l’Energie (ADEME) Direction Villes et Territoires Durables Services Friches Urbaines et Sites Pollués

The behaviour of metals mainly depends on soil p H, carbonate contents and contamination level, which should be considered for the management of contaminated soils. In this study, kitchen garden topsoils(0–25 cm) were sampled from the area around three smelters in France, with different Cd and Pb contamination levels. Effect of a phosphate amendment(a mixture of diammonium phosphate and hydroxyapatite) on the environmental availability and phytoavailability of Cd and Pb was evaluated by different chemical extractions and cultivating lettuce(Lactuca sativa L.), respectively. Changes in the distribution of Cd and Pb were found in most contaminated soils after phosphate amendment. An increase of Cd and Pb in the residual phase was highlighted in almost all carbonated contaminated soils, whereas a decrease of Pb in the exchangeable, water and acid-soluble phase was observed in most contaminated soils with the lowest carbonate contents. The concentrations of extractable Cd and Pb using calcium chloride and acetic and citric acids generally decreased after the soil amendment. Lettuces grown on amended soils were acceptable for human consumption as regard to Pb concentration. In contrast, some lettuces were unacceptable for human consumption, since the concentrations of Cd in the leaves were higher than the European legislation limit. Surprisingly, in carbonated soils with very low concentration of Cd, the Cd concentrations in lettuce reached up to the European legislation limit, making the lettuce unacceptable for human *** study highlighted the fact that the total metal concentration in soils does not always allow to predict the metal accumulation in the edible parts of vegetables in order to make a judgement about their acceptability or unacceptability for human consumption.

关键词： chemical amendment contamination immobilisation lettuce metal urban soil

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Age at reproductive maturity and effect of age and time of day on sex attraction in the potato psyllid Bactericera cockerelli

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Insect Science 2012年第5期19卷 585-594页

作者： christelle Guedot David R. Horton Peter J. Landolt U.S. Department of Agriculture Agricultural Research Service 5230 Konnowac Pass Road Wapato WA 98951 USA

The potato psyllid, Baetericera （= Paratrioza） cockerelli （gulc）（Hemiptera： Triozidae）, is a major pest of potato. Studies were conducted to determine the age at which both males and females reach reproductive maturity and the effect of age and time of day on sex attraction. Adult B. cockerelli reach reproductive maturity within 48 h post-eclosion, with females being mature on the day of eclosion and males at 1 day post-eclosion. Oviposition generally began 2 days after mating but was delayed when females mated within 2 days post-eclosion. In laboratory olfactometer assays, the age of females used as odor sources and the age of males assayed to these odors did not affect sex attraction, with both younger （1-4-day-old） and older （8-10-day-old） males being attracted to females, regardless of female age （1-4-day-old or 8-10-day-old）. Males assayed to live females at different times during the photophase （between 8：00 and 20：00 hours） were attracted to females between 11：00 and 17：00 hours, showing a temporal periodicity in sex attraction with B. cockerelli at least during the photophase.

关键词： mate attraction olfactometer oviposition periodicity potato preference,Triozidae

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From the dilute solution to the pure compound:Extraction strategy based on a multi-stage process of phase separation

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Science China Chemistry 2010年第9期53卷 2070-2076页

作者： GOUTAUDIER christelle TENU Richard COUNIOUX Jean-Jacques Université de Lyon Université Claude Bernard Lyon 1Laboratoire Multimatériaux et Interfaces

It is very rare that a one-step process of extraction leads to the pure compound with a high degree of purity specified by an industrial *** various stages of a synthesis process and possible secondary reactions may lead to the synthesis of more or less complex and highly diluted *** this work,the rationale and strategy for extraction and purification of a high added value compound are *** the thinking is based on the knowledge and the exploitation of phase diagrams and then developed for different unit operations of the *** most significant research tools are the experimental data and the modelling of phase equilibrium to estimate the yield of each step of *** significant example chosen involves all the basic methods of phase separation,starting with liquid-vapour equilibrium:stripping of high volatility components and then more or less complex distillation are classically *** theoretical plateau number can be deduced from the equilibrium equation *** second step is based on the study of the liquid-liquid equilibrium and is an intermediate step for enrichment of the solution when distillation is not possible.A final step based on solid-liquid equilibrium consists of the selective crystallization of the pure product at low temperature,in order to satisfy the requirements of purity and safety imposed by industrial *** conclusion includes all isolation operations in the form of a general extraction and purification scheme.

关键词： phase diagram azeotropic distillation liquid-liquid extraction selective crystallization

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Advances in the engineering of the outer blood-retina barrier:From in-vitro modelling to cellular therapy

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Bioactive Materials 2024年第1期31卷 151-177页

作者： Chloé Dujardin Walter Habeler christelle Monville Didier Letourneur Teresa Simon-Yarza UniversitéParis Cité UniversitéSorbonne Paris NordLaboratory for Vascular Translational Science(LVTS)INSERM-U114875018 ParisFrance INSERM U861 I-StemAFMInstitute for Stem Cell Therapy and Exploration of Monogenic Diseases91100Corbeil-EssonnesFrance U861 I-StemAFMUniversitéParis-SaclayUniversitéD’Evry91100Corbeil-EssonnesFrance CECS Centre D’étude des Cellules Souches91100Corbeil-EssonnesFrance

The outer blood-retina barrier(oBRB),crucial for the survival and the proper functioning of the overlying retinal layers,is disrupted in numerous diseases affecting the retina,leading to the loss of the photoreceptors and ultimately of *** study the oBRB and/or its degeneration,many in vitro oBRB models have been developed,notably to investigate potential therapeutic strategies against retinal ***,to this day,most of these pathologies are untreatable,especially once the first signs of degeneration are *** cure those patients,a current strategy is to cultivate in vitro a mature oBRB epithelium on a custom membrane that is further implanted to replace the damaged native *** a description of the oBRB and the related diseases,this review presents an overview of the oBRB models,from the simplest to the most ***,we propose a discussion over the used cell types,for their relevance to study or treat the *** designed for in vitro applications are then examined,by paying particular attention to the design evolution in the last years,the development of pathological models and the benefits of co-culture models,including both the retinal pigment epithelium and the ***,this review focuses on the models developed for in vivo implantation,with special emphasis on the choice of the material,its processing and its characterization,before discussing the reported pre-clinical and clinical trials.

关键词： Outer-blood retina barrier Tissue engineering and cellular therapy In-vitro disease modelling 3D complex models

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Morphology of living cells cultured on nanowire arrays with varying nanowire densities and diameters

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Science China(Life Sciences) 2018年第4期61卷 427-435页

作者： Zhen Li Henrik Persson Karl Adolfsson Stina Oredsson christelle N. Prinz Division of Solid State Physics Lund University Lund 22100 Sweden Nano Lund Lund University Lund 22100 Sweden Department of Biology Lund 22362 Sweden Present address: Translational & Bioengineering Program Mechanical and Industrial Engineering University of Toronto Toronto MSG 1M1 ON Canada

Vertical nanowire arrays are increasingly investigated for their applications in steering cell behavior. The geometry of the array is an important parameter, which influences the morphology and adhesion of cells. Here, we investigate the effects of array geometry on the morphology of MCF7 cancer cells and MCF10A normal-like epithelial cells. Different gallium phosphide nanowire array-geometries were produced by varying the nanowire density and diameter. Our results show that the cell size is smaller on nanowires compared to flat gallium phosphide. The cell area decreases with increasing the nanowire density on the substrate. We observed an effect of the nanowire diameter on MCF10A cells, with a decreased cell area on 40 nm diameter nanowires, compared to 60 and 80 nm diameter nanowires in high-density arrays. The focal adhesion morphology depends on the extent to which cells are contacting the substrate. For low nanowire densities and diameters, ceils are lying on the substrate and we observed large focal adhesions at the cell edges. In contrast, for high nanowire densities and diameters, cells are lying on top of the nanowires and we observed point-like focal adhesions distributed over the whole cell. Our results constitute a step towards the ability to fine-tune cell behavior on nanowire arrays.

关键词： nanowires cell morphology diameter density aspect ratio

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Use of human pluripotent stem cells to study and treat retinopathies

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World Journal of Stem Cells 2015年第3期7卷 596-604页

作者： Karim Ben M'Barek Florian Regent christelle Monville INSERM UMR861 I-Stem AFM Genopole Campus 1 91030Evry France UEVE UMR861 I-Stem AFM Genopole Campus 1 91030 Evry France

Human cell types affected by retinal diseases(such as age-related macular degeneration or retinitis pimentosa) are limited in cell number and of reduced accessibility. As a consequence, their isolation for in vitro studies of disease mechanisms or for drug screening efforts is fastidious. Human pluripotent stem cells(h PSCs), either of embryonic origin or through reprogramming of adult somatic cells,represent a new promising way to generate models of human retinopathies, explore the physiopathological mechanisms and develop novel therapeutic strategies. Disease-specific human embryonic stem cells were the first source of material to be used to study certain disease states. The recent demonstration that human somatic cells, such as fibroblasts or blood cells, can be genetically converted to induce pluripotent stem cells together with the continuous improvement of methods to differentiate these cells into disease-affected cellular subtypes opens new perspectives to model and understand a large number of human pathologies, including retinopathies. This review focuses on the added value of h PSCs for the disease modeling of human retinopathies and the study of their molecular pathological mechanisms. We also discuss the recent use of these cells for establishing the validation studies for therapeutic intervention and for the screening of large compound libraries to identify candidate drugs.

关键词： Drug screening Human pluripotent stemcells Disease modeling Retinitis pigmentosa

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