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Synthesis and characterization of molecularly imprinted polymers for the selective extraction of oxazepam from complex environmental and biological samples
Fanny Varenne, Porkodi Kadhirvel, Pauline Bosman, Loïc Renault, Audrey Combès, Valérie Pichon
Anal. Bioanal. Chem - - doi: 10.1007/s00216-021-03268-w. - 2021
Oxazepam, one of the most frequently prescribed anxiolytic drugs, is not completely removed from wastewater with conventional treatment processes. It can thus be found at trace levels in environmental water, with human urine constituting the major source of contamination. This study focused on the development and characterization of molecularly imprinted polymers (MIPs) for the selective solid-phase extraction of oxazepam at trace levels from environmental water and human urine samples. Two MIPs were synthesized, and their selectivity in pure organic and aqueous media were assayed. After optimizing the extraction procedure adapted to a large sample volume to reach a high enrichment factor, the most promising MIP was applied to the selective extraction of oxazepam from environmental water. Extraction recoveries of 83 ± 12, 92 ± 4 and 89 ± 10% were obtained using the MIP for tap, mineral and river water, respectively, while a recovery close to 40% was obtained on the corresponding non-imprinted polymer (NIP). Thanks to the high enrichment factors, a limit of quantification (LOQ) of 4.5 ng L-1 was obtained for river water. A selective extraction procedure was also developed for urine samples and gave rise to extraction recoveries close to 95% for the MIP and only 23% for the NIP. Using the MIP, a LOQ of 357 ng L-1 was obtained for oxazepam in urine. The use of the MIP also helped to limit the matrix effects encountered for the quantification of oxazepam in environmental samples and in human urine samples after extraction on an Oasis HLB sorbent.

Identification and semi-relative quantification of intact glycoforms of human chorionic gonadotropin alpha and beta subunits by nano liquid chromatography-Orbitrap mass spectrometry
AmiraAl Matari, Anastasia Goumenou, Audrey Combèsa, Thierry Fournier, Valérie Pichon, Nathalie Delaunay
J. Chromatography A - 1640 461945 - doi.org/10.1016/j.chroma.2021.461945 - 2021
The human chorionic gonadotropin (hCG) protein belongs to a family of glycoprotein hormones called gonadotropins. It is a heterodimer made of two non-covalently linked subunits. The α-subunit structure, hCGα, has 2 N-glycosylation sites, while the beta subunit, hCGβ, has 2 N- and 4 O-glycosylation sites. This leads to numerous glycoforms. A method based on the analysis of hCG glycoforms at the intact level by nano-reversed phase liquid chromatography coupled to high resolution mass spectrometry (nanoLC-HRMS) with an Orbitrap analyzer was previously developed using a recombinant hCG-based drug, Ovitrelle®, as standard. It allowed the detection of about 30 hCGα glycoforms, but didn't allow the detection of hCGβ glycoforms. This method was thus here significantly modified (addition of a pre-concentration step of the sample to increase the sample volume from 70 nl to 1 µl, optimization of the gradient slope and the nature and content of the acidic additive in the mobile phase). It led to an improvement of the separation of hCGα and hCGβ glycoforms, which allowed for the first time the detection of 33 hCGβ glycoforms at intact level. In addition, a higher number of hCGα glycoforms (42 in total, i.e. a 40% increase) was detected. The figures of merit of this new method were next assessed. The relative standard deviations (RSDs) of the retention time ranged between 0.02 and 0.95% (n = 3), with an average value of 0.36% for the alpha glycoforms and between 0.01 and 1.08% (n = 3) with an average value of 0.23% for the beta glycoforms. The RSDs of the relative peak area measured on the extracted ion chromatogram of each glycoform were below 20% (n = 3), with an average value of 9.8%, thus allowing semi-relative quantification. Therefore, this method has a high potential for rapid quality control aiming for the detection and comparison of glycoforms present in glycoprotein-based pharmaceutical preparations.


Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the analysis of tryptic digest of human hemoglobin exposed to sulfur mustard
Florine Hallez, Audrey Combès, Charlotte Desoubries, Anne Bossée, Valérie Pichon
J. Chromatography A - 1163 122518 - doi.org/10.1016/j.jchromb.2020.122518 - 2021
Sulfur mustard is a highly reactive chemical warfare agent that causes severe damages to the victims exposed by alkylating multiple biomolecules such as proteins. Resulting alkylated products can be used as biomarkers of exposure to this chemical agent. A liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method was thus developed to detect alkylated peptides after the tryptic digestion of hemoglobin (50 mg.mL−1) incubated with sulfur mustard at different concentrations (0.25, 0.5, 1, 10 and 100 µg.mL−1). Five new alkylation sites were accurately identified on the protein (α-His72, α-His87, α-His89, β-His2 and β-Val98) and fifteen adducted peptides were detected, among which eight of them resulted from the alkylation of four peptides, each presenting two potential sites of adduction that could be discriminated by the method specificity. Similarly, it was possible to discriminate the three potential adduction sites of the peptide α-T9. Moreover, the method allowed the quantification of all the alkylated peptides with a satisfying repeatability, with RSD ranging from 0.5 to 9.3% for an exposure of hemoglobin to sulfur mustard at 100 µg.mL−1. The analysis of hemoglobin incubated with different concentrations of sulfur mustard levels led to a linear response for all the alkylated peptides with the studied concentrations (0.25, 0.5, 1, 10 and 100 µg.mL−1). A variation of the alkylation rate was also observed between the different peptides studied, with a preferential adduction of sulfur mustard on the histidine residues but also on the N-terminal valine residues of both globin chains and on the Val98 residue of globin β. Furthermore, the presented method proved to be sensitive, with a theoretical possibility to detect alkylated peptides resulting from in vitro incubation of hemoglobin in deionized water with sulfur mustard at 2.63 ng.mL−1. After further development, this method could potentially be used for the analysis of blood samples in vivo exposed to sulfur mustard.



COVID-19 and Dentistry in 72 Questions: An Overview of the Literature
Stéphane Derruau, Jérôme Bouchet, Ali Nassif, Alexandre Baudet, Kazutoyo Yasukawa, Sandrine Lorimier, Isabelle Prêcheur, Agnès Bloch-Zupan, Bernard Pellat, Hélène Chardin
J. Clin. Med. - 10 122518 - 4 - 2021
The outbreak of Coronavirus Disease 2019 (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has significantly affected the dental care sector. Dental professionals are at high risk of being infected, and therefore transmitting SARS-CoV-2, due to the nature of their profession, with close proximity to the patient’s oropharyngeal and nasal regions and the use of aerosol-generating procedures. The aim of this article is to provide an update on different issues regarding SARS-CoV-2 and COVID-19 that may be relevant for dentists. Members of the French National College of Oral Biology Lecturers (“Collège National des EnseignantS en Biologie Orale”; CNESBO-COVID19 Task Force) answered seventy-two questions related to various topics, including epidemiology, virology, immunology, diagnosis and testing, SARS-CoV-2 transmission and oral cavity, COVID-19 clinical presentation, current treatment options, vaccine strategies, as well as infection prevention and control in dental practice. The questions were selected based on their relevance for dental practitioners. Authors independently extracted and gathered scientific data related to COVID-19, SARS-CoV-2 and the specific topics using scientific databases. With this review, the dental practitioners will have a general overview of the COVID-19 pandemic and its impact on their practice. View Full-Text



Molecularly imprinted polymers in miniaturized extraction and separation devices
Thomas Bouvarel, Nathalie Delaunay, Valérie Pichon
First published - 44(8) 1727-1751 - doi.org/10.1002/jssc.202001223 - 2021
Molecularly imprinted polymers are highly selective and cost-effective materials, which have attracted significant interest in various areas such as sample pretreatment and chromatographic and electrophoretic separations. This review aims to present the state of the art concerning the miniaturization of these materials in order to meet the societal demand for reliable, fast, cheap, and solvent/sample saving analyses. The polymerization route specificities for the production of miniaturized molecularly imprinted polymers in capillaries or chip channels, such as open tubular, packed particles, magnetic nanoparticles, and in situ imprinted monoliths, are investigated. Their performances as selective supports in solid phase extraction and as stationary phases in electrochromatography and liquid chromatography, as well as their possible perspectives are discussed.




Reversible Supra-Folding of User-Programmed Functional DNA Nanostructures on Fuzzy Cationic Substrates
Koyomi Nakazawa, Farah El Fakih, Vincent Jallet, Caroline Rossi-Gendron, Marina Mariconti, Léa Chocron, Mafumi Hishida, Kazuya Saito, Mathieu Morel, Sergii Rudiuk, Damien Baigl
Angew Chem Int Ed Engl - - DOI: 10.1002/anie.202101909 - 2021
We report that user-defined DNA nanostructures, such as two-dimensional (2D) origamis and nanogrids, undergo a rapid higher-order folding transition, referred to as supra-folding, into three-dimensional (3D) compact structures (origamis) or well-defined μm-long ribbons (nanogrids), when they adsorb on a soft cationic substrate prepared by layer-by-layer deposition of polyelectrolytes. Once supra-folded, origamis can be switched back on the surface into their 2D original shape through addition of heparin, a highly charged anionic polyelectrolyte known as an efficient competitor of DNA-polyelectrolyte complexation. Orthogonal to DNA base-pairing principles, this reversible structural reconfiguration is also versatile; we show in particular that 1) it is compatible with various origami shapes, 2) it perfectly preserves fine structural details as well as site-specific functionality, and 3) it can be applied to dynamically address the spatial distribution of origami-tethered proteins.
Protein-protected metal nanoclusters as diagnostic and therapeutic platforms for biomedical applications
ImanZare, Daniel M.Chevrier, Anna Cifuentes-Rius, NasrinMoradi, Yunlei Xianyu, SubhadipGhosh, LauraTrapiella-Alfonso, Ye Tian, Alireza Shourangiz-Haghighi, Saptarshi Mukherjee, Kelong Fan, Michael R.Hamblin
Materials Today - - doi.org/10.1016/j.mattod.2020.10.027 - 2021
The use of protein templates for the controlled synthesis of inorganic nanostructures has gained considerable attention in multidisciplinary fields, including electronics, optics, energy, sensing, and biomedicine, owing to their biocompatibility and structural programmability. The possible synergistic combination of protein scaffolds (and other biomolecules/biopolymers) with metal nanoclusters (MNCs) has created a new class of highly photoluminescent nanoprobes and nanodevices. For the first time, we will discuss the different types of protein templates used for MNC preparation with an emphasis on their optoelectronic properties for application. In particular, applications of protein-coated MNCs for chemosensing or biosensing of cancer biomarkers, neurotransmitters, pathogenic microorganisms, biomolecules, pharmaceutical compounds, and immunoassays are discussed in detail herein. Fluorescence-based and multimodal molecular imaging, both in vitro and in vivo based on functional proteins are also covered. Furthermore, we discuss the burgeoning growth of protein-coated MNCs (e.g., gold (Au) and silver (Ag) NCs) to develop synergistic nanotherapeutics with potential biomedical applications in chemotherapy, radiotherapy, photodynamic therapy (PDT), photothermal therapy (PTT), and antibacterial activity, as well as MNC-containing nanocomposites for enhanced bioimaging and controlled drug release. Overall, the proposed review highlights the recent progress, technical challenges and new horizons in this field, and summarizes our understanding of how MNC properties interact with the biological function of protein scaffolds to develop synergistic nanotherapeutics towards clinical translation.
Superparamagnetic iron oxide nanoparticles functionalized with a binary alkoxysilane array and poly(4-vinylpyridine) for magnetic targeting and pH-responsive release of doxorubicin
Fernando Espinola-Portilla,ab Oracio Serrano-Torres, Gilberto F. Hurtado-López, Uriel Sierra, Anne Varenne, Fanny d’Orlyé, Laura Trapiella-Alfonso, Silvia Gutiérrez-Granados and Gonzalo Ramírez-García
New Journal of Chemistry - 45 3600-3609 - doi.org/10.1039/D0NJ05227B - 2021
Chemotherapeutic drugs cause harmful side effects in cancer patients due to their low specificity, calling for the development of more effective strategies for their dosage and administration. In this work, a smart drug nanocarrier was synthesized through the covalent functionalization of superparamagnetic iron oxide nanoparticles with a triblock copolymer, which includes a dual alkoxysilane array, ((3-aminopropyl)triethoxysilane and (trimethoxysilyl)propyl methacrylate), and the pH-responsive poly(4-vinylpyridine). The synthetic conditions were optimized through structural and physicochemical characterization after every functionalization step. Afterward, the systematic loading, capture, and release of the anticancer drug doxorubicin (Dox) were demonstrated at relevant pH values using a specially designed square wave voltammetry technique. This strategy revealed that the P4VP polymeric chains underwent reversible hydrophobic to hydrophilic transitions in acidic media, triggering a molecular distention driven by the induced intermolecular electro-repulsive forces. Thereafter, the Dox solution can easily penetrate the polymeric layer at pH values below 5.62 (the pKa of poly(4-vinylpyridine)), allowing a loading of 61.9 ± 5.4 mg g−1 in the nanocomplex. After deprotonation in a pH 7.4 buffer solution, the polymer chains underwent intermolecular interactions again, capturing the drug molecules. Subsequently, 93.5 ± 3.5% of the payload was released upon suspension of the nanocomplex in pH 4.0 media, which is significantly more acidic than healthy tissues. Since the magnetic properties of the MNPs were practically unaffected by the surface modification, this nanocomplex offers a versatile strategy for the pH-selective and magnetically-guided release of drugs.

Non-amplified impedimetric genosensor for quantification of miRNA-21 based on the use of reduced graphene oxide modified with chitosan
Michael López Mujica Yuanyuan Zhang Fabiana Gutierrez Féthi Bédioui Gustavo Rivasa
Microchemical Journal - 160 105596 - https://doi.org/10.1016/j.microc.2020.105596 - 2021
We report here an impedimetric genosensor for the quantification of microRNA-21 using [Fe(CN)6]3−/4− as redox probe to transduce the hybridization event. The biosensing platform was built at a thiolated-gold electrode by covalent bond of reduced graphene oxide (RGO) modified with chitosan (CHIT) and further covalent attachment of the aminated DNA probe. GO was used to provide the carboxylic groups for the covalent attachment of CHIT and, once reduced, to improve the electroactivity of the resulting platform, while CHIT served as a bridge between the thiol and the aminated probe DNA. The proposed bioanalytical platform allows the label-free, non-amplified, simple and fast biosensing of microRNA-21, with a linear range between 1.0 × 10−12 M and 1.0 × 10−8 M, a sensitivity of (134 ± 4) ΩM−1 (r2 = 0.996), a detection limit of 300 fM, and a reproducibility of 5.9% for 1.0 × 10−12 M miRNA-21 and 2.2% for 1.0 × 10−9 M miRNA-21. The genosensor was successfully used for the quantification of microRNA-21 in enriched human blood serum, urine and saliva samples.


Corrosion analysis of AISI 430 stainless steel in the presence of Escherichia coli and Staphylococcus aureus
C.Guerra A.Ringuedé M.I.Azocar M.Walter C.Galarce F.Bedioui M.Cassir M.Sancy
Corrosion Science - 181 109204 - doi.org/10.1016/j.corsci.2020.109204 - 2021
AISI 430 stainless steel is an attractive material to be used in the healthcare industry, particularly as a sensor due to its low cost, corrosion resistance, as well as being Ni-free. AISI 430 was evaluated in an artificial sweat solution with the presence of Escherichia coli, and Staphylococcus aureus. Surface microbial analyses did not reveal colonization of bacteria on metallic surfaces, even when bacteria adhesion was investigated in a Müeller-Hinton solution. However, by electrochemical techniques, the AISI 430 surfaces demonstrated clear signs of corrosion mainly in a sterile medium after two weeks of exposure.




628 publications.