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Longitudinal Analyses of Blood Transcriptome During Conversion to Psychosis
Laboratoire Physique des biomolécules - Saurabh Raj, Debjani Bagchi, Jessica Valle Orero, Josette Banroques, N Kyle Tanner, Vincent Croquette
Schizophr Bull - 45(1) 247-255 - doi: 10.1093/schbul/sby009 - 2019
The biological processes associated with the onset of schizophrenia remain largely unknown. Current hypotheses favor gene × environment interactions as supported by our recent report about DNA methylation changes during the onset of psychosis. Here, we conducted the first longitudinal transcriptomic analysis of blood samples from 31 at-risk individuals who later converted to psychosis and 63 at-risk individuals who did not. Individuals were followed for a maximum of 1 year. Blood samples were collected at baseline and at the end of follow-up and individuals served as their own controls. Differentially expressed genes between the 2 groups were identified using the RNA sequencing of an initial discovery subgroup (n = 15 individuals). The most promising results were replicated using high-throughput real-time qPCR in the whole cohort (n = 94 individuals). We identified longitudinal changes in 4 brain-expressed genes based on RNAseq analysis. One of these genes (CPT1A) was replicated in the whole cohort. The previously observed hypermethylation in NRP1 and GSTM5 during the onset of psychosis correlated with a decrease in corresponding gene expression. RNA sequencing also identified 2 co-expression networks that were impaired after conversion compared with baseline-the Wnt pathway including AKT1, CPT1A and semaphorins, and the Toll-like receptor pathway, related to innate immunity. This longitudinal study of transcriptomic changes in individuals with at-risk mental state revealed alterations during conversion to psychosis in pathways and genes relevant to schizophrenia. These results may be a first step toward better understanding psychosis onset. They may also help to identify new biomarkers and targets for disease-modifying therapeutic strategies
Longitudinal Analyses of Blood Transcriptome During Conversion to Psychosis
Laboratoire Physique des biomolécules - Saurabh Raj, Debjani Bagchi, Jessica Valle Orero, Josette Banroques, N Kyle Tanner, Vincent Croquette
Schizophr Bull - 45(1) 247-255 - doi: 10.1093/schbul/sby009 - 2019
The biological processes associated with the onset of schizophrenia remain largely unknown. Current hypotheses favor gene × environment interactions as supported by our recent report about DNA methylation changes during the onset of psychosis. Here, we conducted the first longitudinal transcriptomic analysis of blood samples from 31 at-risk individuals who later converted to psychosis and 63 at-risk individuals who did not. Individuals were followed for a maximum of 1 year. Blood samples were collected at baseline and at the end of follow-up and individuals served as their own controls. Differentially expressed genes between the 2 groups were identified using the RNA sequencing of an initial discovery subgroup (n = 15 individuals). The most promising results were replicated using high-throughput real-time qPCR in the whole cohort (n = 94 individuals). We identified longitudinal changes in 4 brain-expressed genes based on RNAseq analysis. One of these genes (CPT1A) was replicated in the whole cohort. The previously observed hypermethylation in NRP1 and GSTM5 during the onset of psychosis correlated with a decrease in corresponding gene expression. RNA sequencing also identified 2 co-expression networks that were impaired after conversion compared with baseline-the Wnt pathway including AKT1, CPT1A and semaphorins, and the Toll-like receptor pathway, related to innate immunity. This longitudinal study of transcriptomic changes in individuals with at-risk mental state revealed alterations during conversion to psychosis in pathways and genes relevant to schizophrenia. These results may be a first step toward better understanding psychosis onset. They may also help to identify new biomarkers and targets for disease-modifying therapeutic strategies
Optical control of protein activity and gene expression by photoactivation of caged cyclofen
Laboratoire Physique des biomolécules - Hamouri F, Zhang W, Aujard I, Le Saux T, Ducos B, Vriz S, Jullien L, Bensimon D
Methods Enzymol - 624 44927 - doi: 10.1016/bs.mie.2019.04.009 - 2019
The use of light to control the expression of genes and the activity of proteins is a rapidly expanding field. While many of these approaches use a fusion between a light activatable protein and the protein of interest to control the activity of the latter, it is also possible to control the activity of a protein by uncaging a specific ligand. In that context, controlling the activation of a protein fused to the modified estrogen receptor (ERT) by uncaging its ligand cyclofen-OH has emerged as a generic and versatile method to control the activation of proteins quantitatively, quickly and locally in a live organism. Here, we present the experimental details behind this approach.
Glutamate spillover in C. elegans triggers repetitive behavior through presynaptic activation of MGL-2/mGluR5
Laboratoire pour la biologie quantitative du développement - Katz M, Corson F, Keil W,Singhal A, Bae A, Lu Y, Liang Y, Shaham S.
Nat Commun - 10(1) 1882 - doi: 10.1038/s41467-019-09581-4. - 2019
Glutamate is a major excitatory neurotransmitter, and impaired glutamate clearance following synaptic release promotes spillover, inducing extra-synaptic signaling. The effects of glutamate spillover on animal behavior and its neural correlates are poorly understood. We developed a glutamate spillover model in Caenorhabditis elegans by inactivating the conserved glial glutamate transporter GLT-1. GLT-1 loss drives aberrant repetitive locomotory reversal behavior through uncontrolled oscillatory release of glutamate onto AVA, a major interneuron governing reversals. Repetitive glutamate release and reversal behavior require the glutamate receptor MGL-2/mGluR5, expressed in RIM and other interneurons presynaptic to AVA. mgl-2 loss blocks oscillations and repetitive behavior; while RIM activation is sufficient to induce repetitive reversals in glt-1 mutants. Repetitive AVA firing and reversals require EGL-30/Gαq, an mGluR5 effector. Our studies reveal that cyclic autocrine presynaptic activation drives repetitive reversals following glutamate spillover. That mammalian GLT1 and mGluR5 are implicated in pathological motor repetition suggests a common mechanism controlling repetitive behaviors.
HLH-2/E2A Expression Links Stochastic and Deterministic Elements of a Cell Fate Decision during C. elegans Gonadogenesis
Laboratoire pour la biologie quantitative du développement - Michelle A.Attner, WolfgangKeil, Justin M.Benavidez, IvaGreenwald
Nat Commun - 29(18) 3094-3100.e4 - https://doi.org/10.1016/j.cub.2019.07.062 - 2019
Stochastic mechanisms diversify cell fate in organisms ranging from bacteria to humans [1, 2, 3, 4]. In the anchor cell/ventral uterine precursor cell (AC/VU) fate decision during C. elegans gonadogenesis, two “α cells,” each with equal potential to be an AC or a VU, interact via LIN-12/Notch and its ligand LAG-2/DSL [5, 6]. This LIN-12/Notch-mediated interaction engages feedback mechanisms that amplify a stochastic initial difference between the two α cells, ensuring that the cell with higher lin-12 activity becomes the VU while the other becomes the AC [7, 8, 9]. The initial difference between the α cells was originally envisaged as a random imbalance from “noise” in lin-12 expression/activity [6]. However, subsequent evidence that the relative birth order of the α cells biases their fates suggested other factors may be operating [7]. Here, we investigate the nature of the initial difference using high-throughput lineage analysis [10]; GFP-tagged endogenous LIN-12, LAG-2, and HLH-2, a conserved transcription factor that orchestrates AC/VU development [7, 11]; and tissue-specific hlh-2 null alleles. We identify two stochastic elements: relative birth order, which largely originates at the beginning of the somatic gonad lineage three generations earlier, and onset of HLH-2 expression, such that the α cell whose parent expressed HLH-2 first is biased toward the VU fate. We find that these elements are interrelated, because initiation of HLH-2 expression is linked to the birth of the parent cell. Finally, we provide a potential deterministic mechanism for the HLH-2 expression bias by showing that hlh-2 is required for LIN-12 expression in the α cells.
A new injection system for spraying liquid nitrates in a low power plasma reactor: Application to local repair of damaged thermal barrier coating
Laboratoire Procédés - Plasmas - Microsystèmes - F.Rousseau A.Quinsac D.Morvan M.-P.Bacos O.Lavigne C.Rio C.Guinard B.Chevillard
ELSEVIER - 357 195-203 - https://doi.org/10.1016/j.surfcoat.2018.09.069 - 2019
In addition to the search for new Thermal Barrier Coating (TBCs) systems with increased reliability over very long time periods, the repair of current systems is a technological and economic issue for both civilian and military engine end-users. This paper describes the latest version of the deposition process known as the Low-Power Plasma Reactor (LPPR) process, specially developed to repair locally damaged TBCs. The LPPR process enables micro/nanostructured TBCs to be made from nitrate salts in aqueous solutions, which are sprayed in an Ar/O2 plasma discharge at low power (240 W) and transformed into oxide coatings. A new injection device was designed to produce a fairly homogenous and reproducible spray to repair partially spalled APS and EB-PVD TBCs deposited on small flat coupons. The microstructure and the stability of the LPPR TBCs were assessed, in particular using SEM observations, during ageing tests under various time/temperature conditions. The Particle Image Velocimetry (PIV) technique and associated modeling have proved that the nitrates impact the substrate in a liquid state even in the presence of plasma and a vacuum. Due to the liquid state of the precursors, the new LPPR TBC seals the damaged areas and deeply infiltrates all porosities and failure cracks in the original coatings. This research has enabled the new version of the LPPR process to be validated as a simple, efficient, cheap and promising way to repair locally damaged TBCs
Plasma-catalytic hybrid process for CO2 methanation: optimization of operation parameters
Laboratoire Procédés - Plasmas - Microsystèmes - M. Mikhail, B. Wang, R. Jalain, S. Cavadias, M. Tatoulian, S. Ognier, M. E. Gálvez & P. Da Costa
Reaction Kinetics, Mechanisms and Catalysis - 126 629–643 - doi.org/10.1007/s11144-018-1508-8 - 2019
The present study focuses on the hybrid plasma catalytic process for CO2 methanation. This plasma-catalytic process, based on the combination of a DBD plasma and Ni/CeZrO2 catalyst, has several advantages over conventional catalysis: it operates at ambient conditions and requires no external heating. An optimization of the process considering the effect of the different operational parameters such as voltage, GHSV, catalyst mass, flow rate, discharge length, is herein presented. Moreover, a spectroscopic study, aiming to understand the mechanism of the reaction, is also showed. At temperatures around 270 °C and under adiabatic conditions, CO2 conversion rates of about 80% were measured, with a CH4 selectivity greater than 95%.
Surface functionalization of cyclic olefin copolymer by plasma‐enhanced chemical vapor deposition using atmospheric pressure plasma jet for microfluidic applications
Laboratoire Procédés - Plasmas - Microsystèmes - Samantha Bourg Sophie Griveau Fanny d'Orlyé Michael Tatoulian Fethi Bedioui Cédric Guyon Anne Varenne
FULL PAPER - 16 6 - doi.org/10.1002/ppap.201800195 - 2019
Lab‐On‐A‐Chips promise solutions for high throughput and specific analysis for environmental and health applications, with the challenge to develop materials allowing fast, easy, and cheap microfabrication and efficient surface treatment. Cyclic olefin copolymer (COC) is a promising thermoplastic, easily microfabricated for both rapid prototyping and low‐cost mass production of microfluidic devices but still needing efficient surface modification strategies. This study reports for the first time the optimization of an easy COC silica coating process by plasma‐enhanced chemical vapor deposition at atmospheric pressure with plasma jet and tetraethylorthosilicate as precursor, leading to a 158 ± 7 nm thickness and a 14‐day‐stability of hydrophilic properties for a COC‐embedded microchannel (100 µm), paving the way for a simplified and controlled COC surface modification.
Degradation of glucocorticoids in aqueous solution by dielectric barrier discharge: Kinetics, mechanisms, and degradation pathways
Laboratoire Procédés - Plasmas - Microsystèmes - Liu YN, Wang CH, Shen X, Zhang A, Yan SW, Li X, Miruka AC, Wu SM, Guo Y, Ognier S
Chemical Engineering Journal - 374 412-428 - DOI10.1016/j.cej.2019.05.154 - 2019
Performance and mechanism of non-thermal plasma (NTP) technology in removing glucocorticoids (GCs) was investigated using a dielectric barrier discharge (DBD) reactor with fluocinolone acetonide (FA), triamcinolone acetonide (TA) and clobetasol propionate (CP) as representative compounds. Effects of discharge power, plasma-working gases, initial pH, coexistence of ions, and various water matrices (ultrapure water, lake water, drinking water, wastewater effluent) on GC removal and energy yield were evaluated. The results confirm that DBD treatment could efficiently remove FA, TA, and CP, achieving efficiency of 72% (k = 0.0126 min(-1)), 71% (k = 0.0096 min(-1)), and 74% (k = 0.0116 min(-1)), respectively in air-DBD system at 45.2 W, with the process following the first order kinetics and energy yield of 6 mg kW(-1) h(-1). The removal efficiency decreased when adding radical scavengers, indicating that hydroxyl radicals played an important role in GC degradation, while other active species (such as solvated electrons (e(aq)(-)), ozone (O-3), hydrogen peroxide (H2O2) and ultraviolet photolysis (UV)) also contribute to GC degradation. The intermediates generated during the process were analyzed using quadrupole time-of-flight mass spectrometry (QTOF-MS). A total of 23 transformation products of FA, TA and CP were identified, and it was noted that substitution of halogen atoms with center dot OH, oxidation of hydroxyl group to keto acid, decarboxylation of the keto acid, addition of center dot OH, intramolecular cyclization, and hydrolysis of esters occurred during GC degradation by DBD treatment.
Plasma Polymer Layers with Primary Amino Groups for Immobilization of Nano- and Microparticles
Laboratoire Procédés - Plasmas - Microsystèmes - Xi Rao; Ali Abou Hassan; Cédric Guyon; Mengxue Zhang; Stephanie Ognier; Michaël Tatoulian
Plasma Chemistry and Plasma Processing - 2 178 - DOI: 10.1007/s11090-019-10056-z - 2019
The assembly of nano- and micro-scale building blocks on surface has been the focus of intense interest in materials science for years. In this work, (3-aminopropyl)triethoxysilane (APTES) carrying one primary amino group was deposited on various substrate surfaces using the plasma polymerization method. The key plasma parameters i.e. pressure and power were varied to obtained the highest density of primary amino groups. The influence of such parameters on the characteristics of deposited layers (e.g. chemical structure, adhesion strength, growth rate, etc.) was systemically investigated using various characterization methods such as XPS, FTIR, ellipsometry and so on. Meanwhile, three types of particles (AuNPs, zeolites and gold@zeolites) with sizes from nano- to submicro-range were synthesized and further used as model building blocks. Subsequently, the prepared particles were deposited onto cyclic olefin copolymer (COC) substrate surfaces, which were pre-functionalized by deposition of the plasma polymer layer using the parameters of pressure = 1.0 mbar and power = 30 W. The results confirmed the formation of membrane structures consisting of highly packed particles on the COC surface, and such immobilized structures showed high stability against flowing water, evidencing the good immobilization ability of deposited APTES layers with amino groups.
Thermo-mechanical and photo-luminescence properties of micro-actuators made of liquid crystal elastomers with cyano-oligo(p-phenylene vinylene) crosslinking bridges
Laboratoire Procédés - Plasmas - Microsystèmes - Bin Ni, Hui Chen, Mengxue Zhang, Patrick Keller, Michael Tatoulian and Min-Hui Li
Materials Chemisty Frontiers - 3 2499-2506 - https://doi.org/10.1039/C9QM00480G - 2019
Nematic liquid crystal elastomer (LCE) micropillars with reversible thermomechanical deformations and photo luminescence (PL) intensity variations were successfully fabricated by introducing a cyano-oligo(p-phenylene vinylene) dye as a chemical crosslinker. The PL intensity of the micropillars decreased and increased reversibly during the thermal-deformation process. We studied in detail the possible factors that influence the PL intensity variations of the micropillars, including temperature variation, contraction/extension and phase transition. The dye molecules mainly kept the “monomer” state in the micropillars during the thermo-activated deformation. It was found that the phase transition from nematic to isotropic of the LCEs played the major role in the PL intensity variations. This kind of micropillar may have potential application in fluorescent soft sensors and actuators.

Synthesis of benzaldehyde with high selectivity using immobilized AuNPs and AuNPs@zeolite in a catalytic microfluidic system
Laboratoire Procédés - Plasmas - Microsystèmes - Xi Rao, Ali Abou Hassan, Cédric Guyon, Erick Osvaldo Martinez Ruiz, Michaël Tatoulian and Stephanie Ognier
Lab. Chip - 19 2866-2873 - https://doi.org/10.1039/C9LC00386J - 2019
In the present work, gold based catalysts were synthesized and immobilized on the surface of cyclic olefin copolymer (COC) microreactors. The microreactors were subsequently applied in a homemade microfluidic system for synthesizing benzaldehyde by oxidation of benzyl alcohol in water medium. The Au nanoparticles (NPs) immobilized on the inner surface of the microchannel showed a very high selectivity (94%) for benzaldehyde, while zeolite NPs exhibited only an adsorption feature to this reaction. Moreover, the results showed that the AuNP catalytic activity was maintained for at least 9 hours. However, the obtained conversion with AuNPs was only 20%, indicating a relatively low productivity. In comparison, AuNPs assembled on the surface of zeolite NPs (AuNPs@zeolite) and immobilized in the microchannel showed the best catalytic performance, as the highest benzaldehyde selectivity (>99%) with a relatively high benzyl alcohol conversion of 42.4% was achieved under the same conditions. To the best of our knowledge, this is the first example demonstrating the use of AuNP or AuNP@zeolite catalysts in a microsystem performing such high selectivity for benzaldehyde in water medium.

Cross coupling of alkylsilicates with acyl chlorides via photoredox/nickel dual catalysis: a new synthesis method for ketones
Laboratoire Procédés - Plasmas - Microsystèmes - Etienne Levernier, Vincent Corcé, Louise-Marie Rakotoarison, Adrien Smith, Mengxue Zhang, Stephanie Ognier, Michael Tatoulian, Cyril Ollivier and Louis Fensterbank
Organic Chemistry Frontiers - 6 1378-1382 - https://doi.org/10.1039/C9QO00092E - 2019
Photoredox/nickel dual catalysis using easily oxidized bis-catecholato hypercoordinated silicon derivatives as radical sources and acyl chlorides as electrophiles allows a new method of formation of dialkyl and alkyl-aryl ketones as well as dibenzyl ketones which are less easily accessed. Flow chemistry can be used.
Modified UNIFAC (Dortmund) Parameters for the Interaction between the Amino Group at Cycloaliphatic Hydrocarbon and the Hydroxyl Group
Laboratoire Procédés - Plasmas - Microsystèmes - Mandy Klauck, Sandra Richter, Thomas Hähnel, Jürgen Schmelzer, Grit Kalies
X.MOL - 11-12 - DOI: 10.1021/acs.iecr.9b03971 - 2019
Modified UNIFAC (Dortmund) parameters for the interactions of the amino group at cycloaliphatic hydrocarbon with the hydroxyl group or the methanol group were determined by means of literature and our own experimental data. The vapor–liquid equilibria of the three binary systems cyclohexylamine + 2-butanol, cyclohexylamine + 2-methyl-2-butanol, and cyclohexylamine + 1-hexanol were measured isothermally with the dynamic method. The predictions of phase equilibrium data using the new parameter set for the amino group at cycloaliphatic hydrocarbon are compared to the predictions using the amino group at aliphatic hydrocarbon. The improvement of the description of experimental data is shown for binary and ternary systems.
A Comparison Study of Functional Groups (Amine vs. Thiol) for Immobilizing AuNPs on Zeolite Surface
Laboratoire Procédés - Plasmas - Microsystèmes - Xi Rao, Michaël Tatoulian, Cédric Guyon, Stephanie Ognier, Chenglin Chu, and Ali Abou Hassan
Nanomaterials - 9(7) 1034 - doi: 10.3390/nano9071034 - 2019
Immobilization of gold nanoparticles (AuNPs) on the surface of zeolite has received a great interest due to Au@zeolite’s unique characteristics and high performance for catalysis. In this work we studied the grafting of two different functional molecules; one having an amine group (3-aminopropyl)triethoxysilane (APTES) and the second having a thiol group (3-mercaptopropyl)trimethoxysilane (MPTES) on the surface of zeolite using the same wet chemistry method. The modified zeolite surfaces were characterized using zeta potential measurements; diffuse reflectance infrared fourier transform (DRIFT) and X-ray photoelectron spectroscopy (XPS). The results confirmed a successful deposition of both functional groups at the topmost surface of the zeolite. Furthermore; transmission electron microscopy (TEM), ultraviolet-visible (UV-Vis) spectroscopy and XPS results clearly evidenced that APTES provided a better AuNPs immobilization than MPTES as a result of; (1) less active functions obtained after MPTES deposition, and (2) the better attaching ability of thiol to the gold surface.
Two-step local functionalization of fluoropolymer Dyneon THV microfluidic materials by scanning electrochemical microscopy combined to click reaction
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Kadhirvel P, Combès A, Bordron L, Pichon V
Anal. Bioanal. Chem - 411(8) 1525-1536 - doi: 10.1007/s00216-019-01586-8 - 2019
A molecularly imprinted polymer (MIP) was designed in order to allow the selective solid-phase extraction of carbamazepine (CBZ), an anticonvulsant and mood-stabilizing drug, at ultra-trace level from aqueous environmental samples. A structural analog of CBZ was selected as a dummy template and different synthesis conditions were screened. The selectivity of the resulting imprinted polymers was evaluated by studying the retention of CBZ in a solvent similar to the one used for the synthesis. The presence of imprinted cavities in the polymers was then demonstrated by comparing the elution profiles (obtained by using MIP and a non-imprinted polymer, NIP, as a control) of the template, of CBZ, and of a structural analog of CBZ. Then, the extraction procedure was further optimized for the treatment of aqueous samples on the two most promising MIPs, with special attention being paid to the volume and composition of the percolation and washing solutions. The best MIP provided a highly selective retention in tap water with 81% extraction recovery for CBZ in the elution fraction of the MIP and only 14% for NIP. The repeatability of the extraction procedure was demonstrated for both tap and river waters (RSD below 4% in river water) for the drugs CBZ, oxcarbamazepine, and one metabolite (carbamazepine 10,11-epoxide). A MIP capacity of 1.15 μmol g-1 was determined. Finally, an analytical procedure involving the MIP was developed allowing the detection of CBZ at a concentration level of only a few nanograms per liter in river water. The selectivity provided by the MIP resulted in a 3000-fold increase of the signal-to-noise ratio in LC/MS analysis as compared to the use of conventional sorbent. Graphical abstract.
First profiling in hydrophilic interaction liquid chromatography of intact human chorionic gonadotropin isoforms.
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Camperi J, Pichon V, Fournier T, Delaunay N
J Pharm Biomed Anal - 10;174 495-499. - doi: 10.1016/j.jpba.2019.06.014 - 2019
The study of glycoproteins is a rapidly growing field, which is not surprising considering that approximately 70% of human proteins are glycosylated and that numerous biological functions are associated to the glycosylation. In this work, our interest focused on the heterodimeric human Chorionic Gonadotropin (hCG) glycoprotein that is the specific hormone of the human pregnancy, consisting of an α and a β subunit, so-called hCGα and hCGβ, respectively. This protein possesses a very high structural heterogeneity, essentially due to the presence of 8 glycosylation sites, but also other types of post-translational modifications. In this study, for the first time, the potential of hydrophilic interaction liquid chromatography (HILIC) was investigated to separate the intact hCG isoforms. Three different HILIC stationary phases were tested using an hCG-based drug as standard, a recombinant hCG. For each stationary phase, the effect of the initial mobile phase composition based on ACN/H2O mixture, the slope of the gradient, the content and nature of the acidic additive (formic acid and trifluoroacetic acid (TFA)), and the addition of a volatile salt (ammonium formate) on the retention and the resolution were studied. The best HILIC separation was obtained with the amide column and a mobile phase composed of water/ACN containing 0.1% of TFA. The repeatability in terms of retention times and peak areas was then assessed. Finally, the method was applied to the analysis of a second hCG-based drug obtained from urine of pregnant women. Both drugs gave chromatograms with more than 10 peaks. However, they were significantly different, which demonstrated the potential of HILIC method for hCG isoform fingerprinting
Specificity of the metabolic signatures of fish from cyanobacteria rich lakes
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Benoît Sotton, Alain Paris, Séverine Le Manach, Alain Blond, Charlotte Duval, Qin Qiao, Arnaud Catherine, Audrey Combes, Valérie Pichon, Cécile Bernard, Benjamin Marie
Chemosphere - - DOI: 10.1016/j.chemosphere.2019.03.115 - 2019
With the increasing impact of the global warming, occurrences of cyanobacterial blooms in aquatic ecosystems are becoming a main worldwide ecological concern. Due to their capacity to produce potential toxic metabolites, interactions between the cyanobacteria, their cyanotoxins and the surrounding freshwater organisms have been investigated during the last past years. Non-targeted metabolomic analyses have the powerful capacity to study simultaneously a high number of metabolites and thus to investigate in depth the molecular signatures between various organisms encountering different environmental scenario, and potentially facing cyanobacterial blooms.

In this way, the liver metabolomes of two fish species (Perca fluviatilis and Lepomis gibbosus) colonizing various peri-urban lakes of the Île-de-France region displaying high biomass of cyanobacteria, or not, were investigated. The fish metabolome hydrophilic fraction was analyzed by 1H NMR analysis coupled with Batman peak treatment for the quantification and the annotation attempt of the metabolites. The results suggest that similar metabolome profiles occur in both fish species, for individuals collected from cyanobacterial blooming lakes compared to organism from non-cyanobacterial dominant environments. Overall, such environmental metabolomic pilot study provides new research perspectives in ecology and ecotoxicology fields, and may notably provide new information concerning the cyanobacteria/fish ecotoxicological interactions.
First characterizations by capillary electrophoresis of human Chorionic Gonadotropin at the intact level
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Benoît Sotton, Alain Paris, Séverine Le Manach, Alain Blond, Charlotte Duval, Qin Qiao, Arnaud Catherine, Audrey Combes, Valérie Pichon, Cécile Bernard, Benjamin Marie
Talanta - 193 77-86 - doi: 10.1016/j.talanta.2018.09.095 - 2019
In the present work, the first characterizations by Capillary Electrophoresis of the human Chorionic Gonadotropin (hCG) hormone at the intact level were carried out. hCG is a hetero-dimeric glycoprotein, specific to the human pregnancy, consisting of an α and a β subunit, so-called hCGα and hCGβ, respectively. hCG has 8 potential glycosylation sites leading to a high number of isoforms (including glycoforms and other post-translational modifications) that we are interesting to characterize. First, Capillary Gel Electrophoresis (CGE) was used to separate the isoforms of two hCG-based drugs: Ovitrelle® (recombinant r-hCG) and Pregnyl (hCG isolated from the urine of pregnant women, u-hCG). As expected, CGE led to a better resolution than SDS-PAGE and confirmed the large heterogeneity of hCG. Different CGE profiles were obtained for the two hCG-based drugs, varying in number of peaks, migration times, and peak intensities, thus demonstrating that the drugs contain isoforms, different in nature and proportion. This result was confirmed by Capillary IsoElectrophoretic Focusing (CIEF). The pI ranges of the hCG isoforms were found between 3.4 and 4.7, and 4.5 and 5.2 for r-hCG and u-hCG, respectively. This information was further used to develop the separation of the hCG isoforms by Capillary Zone Electrophoresis (CZE). The pH, the nature, and the concentration of the background electrolyte as well as the nature and the content of its organic modifier were optimized. The use of a coated capillary to avoid protein adsorption was also evaluated. The final CZE-UV method allowed distinguishing at least 6 peaks, corresponding to different hCG isoforms. To significantly improve the level of information obtained, the CZE instrument was then coupled by means of an electrospray ionization source to a triple quadrupole (TQ) mass spectrometer. Two detection strategies were used, one focusing on the lower m/z values (100-1000) in order to identify some sugar residues as diagnostic ions to confirm the presence of glycan chains, and the second focusing on the higher m/z values (1000-2000), corresponding to the multiple charged intact protein isoforms. For both approaches, the fragmentor and capillary voltage values were optimized. The composition and the flow-rate of the sheath liquid were then optimized for the strategy focusing on the higher m/z values in order to both increase the charge state of the ionized intact isoforms and the signal-to-noise ratio. The final method was used to compare the two hCG-based drugs, demonstrating the potential of the developed CZE-MS method for isoforms fingerprinting.
Development and application of water-compatible molecularly imprinted polymers for the selective extraction of carbamazepine from environmental waters
Laboratoire Sciences Analytiques Bioanalytiques et Miniaturisation - Porkodi Kadhirvel, Audrey Combès, Louis Bordron, Valérie Pichon
Anal. Bioanal. Chem - 11(8) 1525-1536 - doi: 10.1007/s00216-019-01586-8 - 2019
A molecularly imprinted polymer (MIP) was designed in order to allow the selective solid-phase extraction of carbamazepine (CBZ), an anticonvulsant and mood-stabilizing drug, at ultra-trace level from aqueous environmental samples. A structural analog of CBZ was selected as a dummy template and different synthesis conditions were screened. The selectivity of the resulting imprinted polymers was evaluated by studying the retention of CBZ in a solvent similar to the one used for the synthesis. The presence of imprinted cavities in the polymers was then demonstrated by comparing the elution profiles (obtained by using MIP and a non-imprinted polymer, NIP, as a control) of the template, of CBZ, and of a structural analog of CBZ. Then, the extraction procedure was further optimized for the treatment of aqueous samples on the two most promising MIPs, with special attention being paid to the volume and composition of the percolation and washing solutions. The best MIP provided a highly selective retention in tap water with 81% extraction recovery for CBZ in the elution fraction of the MIP and only 14% for NIP. The repeatability of the extraction procedure was demonstrated for both tap and river waters (RSD below 4% in river water) for the drugs CBZ, oxcarbamazepine, and one metabolite (carbamazepine 10,11-epoxide). A MIP capacity of 1.15 μmol g-1 was determined. Finally, an analytical procedure involving the MIP was developed allowing the detection of CBZ at a concentration level of only a few nanograms per liter in river water. The selectivity provided by the MIP resulted in a 3000-fold increase of the signal-to-noise ratio in LC/MS analysis as compared to the use of conventional sorbent. Graphical abstract.

638 publications.