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14 results on '"Jean-Pierre Alcaraz"'

1. Nanostructural Characterization of Cardiolipin-Containing Tethered Lipid Bilayers Adsorbed on Gold and Silicon Substrates for Protein Incorporation

Author

Jean-Pierre Alcaraz, Donald K. Martin, Andrew Nelson, Giovanna Fragneto, Marco Maccarini, Sebastian Köhler, Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-SyNaBi ), Translational Innovation in Medicine and Complexity / Recherche Translationnelle et Innovation en Médecine et Complexité - UMR 5525 (TIMC ), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), and Université Grenoble Alpes (UGA)

Subjects
Silicon, Cardiolipins, Lipid Bilayers, 02 engineering and technology, 03 medical and health sciences, chemistry.chemical_compound, Electrochemistry, Cardiolipin, Molecule, General Materials Science, Lipid bilayer, Spectroscopy, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Bilayer, Surfaces and Interfaces, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Nanostructures, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Membrane, chemistry, Membrane protein, Biophysics, lipids (amino acids, peptides, and proteins), Gold, Neutron reflectometry, 0210 nano-technology, Ethylene glycol
Abstract

A key to the development of lipid membrane-based devices is a fundamental understanding of how the molecular structure of the lipid bilayer membrane is influenced by the type of lipids used to build the membrane. This is particularly important when membrane proteins are included in these devices since the precise lipid environment affects the ability to incorporate membrane proteins and their functionality. Here, we used neutron reflectometry to investigate the structure of tethered bilayer lipid membranes and to characterize the incorporation of the NhaA sodium proton exchanger in the bilayer. The lipid membranes were composed of two lipids, dioleoyl phosphatidylcholine and cardiolipin, and were adsorbed on gold and silicon substrates using two different tethering architectures based on functionalized oligoethylene glycol molecules of different lengths. In all of the investigated samples, the addition of cardiolipin caused distinct structural rearrangement including crowding of ethylene glycol groups of the tethering molecules in the inner head region and a thinning of the lipid tail region. The incorporation of NhaA in the tethered bilayers following two different protocols is quantified, and the way protein incorporation modulates the structural properties of these membranes is detailed.

Published
2021

2. Controlled Heat and Humidity-Based Treatment for the Reuse of Personal Protective Equipment: A Pragmatic Proof-of-Concept to Address the Mass Shortage of Surgical Masks and N95/FFP2 Respirators and to Prevent the SARS-CoV2 Transmission

Author

Louis Bernard, Guillaume Desoubeaux, Elsa Bodier-Montagutelli, Jeoffrey Pardessus, Déborah Brea, Laurine Allimonnier, Sébastien Eymieux, Pierre-Ivan Raynal, Virginie Vasseur, Laurent Vecellio, Ludovic Mathé, Antoine Guillon, Philippe Lanotte, Jérémie Pourchez, Paul O. Verhoeven, Stéphane Esnouf, Muriel Ferry, Nicolas Eterradossi, Yannick Blanchard, Paul Brown, Philippe Roingeard, Jean-Pierre Alcaraz, Philippe Cinquin, Mustapha Si-Tahar, Nathalie Heuzé-Vourc'h, Service de Médecine Interne et Maladies Infectieuses [Tours], Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Université de Tours, Service de parasitologie, mycologie, médecine tropicale [CHRU Tours], Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), Centre d’Etude des Pathologies Respiratoires (CEPR), UMR 1100 (CEPR), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de biologie cellulaire - Microscopie électronique, Université de Tours-Programme PluriFormation Analyse des Systèmes Biologiques (PPF ASB), Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH - U1259 Inserm - CHRU Tours ), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), Service de Médecine Intensive Réanimation [Tours], Service de bactériologie et d'hygiène hospitalière [Tours], Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Infectiologie et Santé Publique (UMR ISP), Université de Tours-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Mines Saint-Etienne, Univ Lyon, Univ Jean Monnet, INSERM, U 1059 Sainbiose, Centre CIS, F - 42023 Saint-Etienne France, Groupe Immunité des Muqueuses et Agents Pathogènes (GIMAP), Université Jean Monnet [Saint-Étienne] (UJM), Laboratory of Infectious Agents and Hygiene [Saint-Etienne], Centre Hospitalier Universitaire de Saint-Etienne (CHU de Saint-Etienne)-Université Jean Monnet [Saint-Étienne] (UJM), Service d'Etudes du Comportement des Radionucléides (SECR), Département de Physico-Chimie (DPC), CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire de Ploufragan-Plouzané-Niort [ANSES], Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA), Université de Tours (UT), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours (UT)-Programme PluriFormation Analyse des Systèmes Biologiques (PPF ASB), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Chanteloup, Nathalie Katy, Université Jean Monnet - Saint-Étienne (UJM), Centre Hospitalier Universitaire de Saint-Etienne (CHU de Saint-Etienne)-Université Jean Monnet - Saint-Étienne (UJM), Santé Ingénierie Biologie Saint-Etienne (SAINBIOSE), Centre Ingénierie et Santé (CIS-ENSMSE), École des Mines de Saint-Étienne (Mines Saint-Étienne MSE), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-École des Mines de Saint-Étienne (Mines Saint-Étienne MSE), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service des Agents Infectieux et Hygiène [CHU Saint-Etienne], and Centre Hospitalier Universitaire de Saint-Etienne [CHU Saint-Etienne] (CHU ST-E)-Université Jean Monnet - Saint-Étienne (UJM)

Subjects
0301 basic medicine, SARS–CoV-2, business.product_category, Coronavirus disease 2019 (COVID-19), Computer science, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), facemask, [SDV]Life Sciences [q-bio], 030231 tropical medicine, 030106 microbiology, coronavirus, Economic shortage, heating, Reuse, Routine practice, 03 medical and health sciences, 0302 clinical medicine, FFP2/N95, CoV-2, recyclibility, FFP2, Respirator, Personal protective equipment, Original Research, surgical face masks, lcsh:R5-920, N95, COVID-19, General Medicine, 3. Good health, [SDV] Life Sciences [q-bio], Risk analysis (engineering), 13. Climate action, Proof of concept, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, Medicine, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, lcsh:Medicine (General), SARS&#8211
Abstract

International audience; Background: The coronavirus infectious disease-2019 (COVID-19) pandemic has led to an unprecedented shortage of healthcare resources, primarily personal protective equipment like surgical masks, and N95/filtering face piece type 2 (FFP2) respirators.Objective: Reuse of surgical masks and N95/FFP2 respirators may circumvent the supply chain constraints and thus overcome mass shortage. Methods, design, setting, and measurement: Herein, we tested the effects of dry- and moist-air controlled heating treatment on structure and chemical integrity, decontamination yield, and filtration performance of surgical masks and FFP2 respirators.Results: We found that treatment in a climate chamber at 70 degrees C during 1 h with 75% humidity rate was adequate for enabling substantial decontamination of both respiratory viruses, oropharyngeal bacteria, and model animal coronaviuses, while maintaining a satisfying filtering capacity.Limitations: Further studies are now required to confirm the feasibility of the whole process during routine practice.Conclusion: Our findings provide compelling evidence for the recycling of pre-used surgical masks and N95/FFP2 respirators in case of imminent mass shortfall.

Published
2020

3. Reflections on Emerging Technologies in Nanomedicine

Author

Donald K. Martin, Gary T. M. Henehan, Adam Mechler, Ratnesh Lal, Jean-Pierre Alcaraz, Satya Prakash, Munis Dundar, Medicine Faculty, Erciyes University, School of Chemistry, Monash UniVersity, Clayton, VIC. 3800 (Australia), Monash University [Clayton], Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA), Technological University [Dublin] (TU), University of California [San Diego] (UC San Diego), University of California, and Biomedical Technology and Cell Therapy Research Laboratory, Montreal

Subjects
0106 biological sciences, Modern medicine, Bibliometric analysis, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Web of science, Emerging technologies, microbiome, 01 natural sciences, bibliometric analysis, antimicrobial peptides, 03 medical and health sciences, Medicine, Biomedicine, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, lcsh:R5-920, 0303 health sciences, bioinspired technologies, gut microbiota, business.industry, gene-based biomedicine, 3. Good health, Nanomedicine, Engineering ethics, lcsh:Medicine (General), business, 010606 plant biology & botany
Abstract

Nanomedicine technologies seem to play an increasing role in modern medicine. Although there are many challenges, it is important to consider emerging technologies that may impact nanomedicine. To establish a rational basis for such consideration in this review, we summarise the present state-of-the-art in nanomedicine based on a bibliometric analysis of the literature that is available in the Clarivate (R) Web of Science. Then, we build upon that state-of-the-art to highlight the emerging technologies that may have an impact on nanomedicine. We also reflect on these emerging technologies by highlighting examples in areas, such as gene-based biomedicine, the rational design of antimicrobial peptides, and diagnosis and therapy of the gut microbiota. In general, emerging technologies that are based on bioinspired and biomimetic approaches appear to provide important pointers to future improvements in nanomedicine.

Published
2020

4. Functional Characterization of Cell-Free Expressed OprF Porin from Pseudomonas aeruginosa Stably Incorporated in Tethered Lipid Bilayers

Author

Landry Gayet, Lavinia Liguori, Jean-Pierre Alcaraz, Jean-Luc Lenormand, Erik B. Watkins, Marco Maccarini, Barry Stidder, Donald K. Martin, Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Los Alamos National Laboratory (LANL), and Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Protein Conformation, Lipid Bilayers, Porins, Virulence, medicine.disease_cause, Biophysical Phenomena, law.invention, 03 medical and health sciences, law, Electrochemistry, medicine, General Materials Science, Lipid bilayer, ComputingMilieux_MISCELLANEOUS, Spectroscopy, biology, Chemistry, Pseudomonas aeruginosa, Surfaces and Interfaces, Condensed Matter Physics, biology.organism_classification, 030104 developmental biology, Biochemistry, Porin, Recombinant DNA, Neutron reflectometry, Function (biology), Bacteria
Abstract

OprF has a central role in Pseudomonas aeruginosa virulence and thus provides a putative target for either vaccines or antibiotic cofactors that could overcome the bacterium's natural resistance to antibiotics. Here we describe a procedure to optimize the production of highly pure and functional OprF porins that are then incorporated into a tethered lipid bilayer. This is a stable biomimetic system that provides the capability to investigate structural aspects and function of OprF using and neutron reflectometry and electrical impedance spectroscopy. The recombinant OprF produced using the optimized cell-free procedure yielded a quantity of between 0.5 to 1.0 mg/mL with a purity ranging from 85 to 91% in the proteoliposomes. The recombinant OprF is capable of binding IFN-γ and is correctly folded in the proteoliposomes. Because OprF proteins form pores the biomimetic system allowed the measurement of OprF conductance using impedance spectroscopy. The neutron reflectometry measurements showed that the OprF protein is incorporated into the lipid bilayer but with parts of the protein in both the regions above and below the lipid bilayer. Those structural aspects are coherent with the current assumed structure of a transmembrane N-terminal domain composed by eight stranded beta-barrels and a globular C-terminal domain located in the periplasm. Currently there are no crystal structures available for OprF. The experimental model system that we describe provides a basis for further fundamental studies of OprF and particularly for the ongoing biotechnological development of OprF as a target for antibacterial drugs.

Published
2017

5. Cell-free production of VDAC directly into liposomes for integration with biomimetic membrane systems

Author

Jean-Luc Lenormand, Lavinia Liguori, Donald K. Martin, Jean-Pierre Alcaraz, Philippe Cinquin, Barry Stidder, Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Voltage-dependent anion channel, Mitochondrial intermembrane space, Blotting, Western, membrane proteins, Nanotechnology, Biochemistry, Transport Pathway, VDAC liposomes, 03 medical and health sciences, 0302 clinical medicine, Biomimetics, Voltage-Dependent Anion Channels, cell-free expression system, Cloning, Molecular, Microscopy, Immunoelectron, Liposome, Cell-Free System, Biomimetic devices, biology, Chemistry, Circular Dichroism, Membranes, Artificial, General Medicine, electrical measurements, Transport protein, Cytosol, 030104 developmental biology, Membrane protein, Liposomes, Biomimetic Devices, biology.protein, Biophysics, 030217 neurology & neurosurgery, Biotechnology
Abstract

International audience; The mitochondrial voltage-dependent anion channel (VDAC) is a pivotal protein since it provides the major transport pathway between the cytosol and the mitochondrial intermembrane space and it is implicated in cell apoptosis by functioning as a gatekeeper for the trafficking of mitochondrial death molecules. VDAC is a beta-barrel channel with a large conductance, and we use it as a model transport protein for the design of biomimetic systems. To overcome the limitations of classical overexpression methods for producing and purifying membrane proteins (MPs) we describe here the use of an optimized cell-free system. In a one-step reaction VDAC is obtained directly integrated into liposomes and purified by ultracentrifugation. We then combine proteoliposomes with different bilayers models in order to validate VDAC insertion and functionality. This VDAC biomimetic model is the first example validating the use of a cell-free expression system for production of MPs into liposomes and tethered bilayers as a toolbox to build a wide range of biomimetic devices.

Published
2016

6. A Biomimetic Lipid Membrane Device Reveals the Interaction of Cancer Biomarkers with Human Serum Lipidic Moieties

Author

Abir Khalil-Mgharbel, Paul K. Dembélé, Isabelle Vilgrain, Jean-Pierre Alcaraz, Md. Mehadi Hasan Sohag, Donald K. Martin, Helena Polena, Laboratoire de développement et vieillissement de l'endothélium, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut National de la Santé et de la Recherche Médicale (INSERM)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF), and Alcaraz, Jean-Pierre

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Lipoproteins, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, [SDV]Life Sciences [q-bio], Population, [SDV.CAN]Life Sciences [q-bio]/Cancer, Applied Microbiology and Biotechnology, 03 medical and health sciences, 0302 clinical medicine, [SDV.CAN] Life Sciences [q-bio]/Cancer, Antigens, CD, Biomimetic Materials, Biomarkers, Tumor, medicine, Humans, Immunoprecipitation, Nanotechnology, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, education, Lipid bilayer, ComputingMilieux_MISCELLANEOUS, education.field_of_study, Apolipoprotein A-I, biology, Chemistry, Cancer, General Medicine, Models, Theoretical, Cadherins, medicine.disease, Kidney Neoplasms, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, 3. Good health, [SDV] Life Sciences [q-bio], Cholesterol, HEK293 Cells, 030104 developmental biology, Biochemistry, 030220 oncology & carcinogenesis, biology.protein, Molecular Medicine, Biomarker (medicine), lipids (amino acids, peptides, and proteins), Apolipoprotein A1, Cancer biomarkers, Ultracentrifuge, Biotechnology, Lipoprotein
Abstract

A major problem for the detection of cancer biomarkers in plasma or serum is that common clinical practice does not require the patient to be in a fasting state. Considering that lipoproteins are the main population affected by food intake, the authors hypothesized that biomarkers could be embedded in lipid particles and thereby opens a new avenue for detection. Using the recently published biomarker, soluble VE-cadherin (sVE), the authors tested our hypothesis using techniques of biophysics, biochemistry and the tools of nanobiotechnology on serum samples from kidney cancer patients (n = 106). Optical density as well as contact angle measurements of serum revealed heterogeneity in the particle content of the serum samples. Isolation of the lipidic moieties by ultracentrifugation showed that sVE was detected in this compartment. Further, isolation of lipoprotein subclasses by precipitation with sodium phosphotungstate and MgCl2 , showed that HDL carried the majority of sVE. Immunoprecipitation of sVE confirmed that it was associated with Apolipoprotein A1, a major compound of HDL. Using a biomimetic lipid bilayer membrane coupled with impedance spectroscopy the authors quantified, in real-time, that the sVE adsorbed to the lipid bilayer membrane without altering its structure. Taken together, these results show for the first time a direct interaction of a cancer biomarker with lipids. The authors anticipate these results to prompt fasting for future blood tests for large-scale studies in the biomarkers research field.

Published
2018

7. 3D polyelectrolyte scaffolds to mimic exocrine glands: a step towards a prostate-on-chip platform

Author

Odile Filhol-Cochet, Frédérique Kermarrec, Maxime Legues, Xavier Gidrol, Guillaume Laffitte, Christophe A. Marquette, Jean-Pierre Alcaraz, Olivier Nicoud, Nathalie Picollet-D'hahan, Florence Rivera, Isabelle Vilgrain, Lavinia Liguori, Jacques Thélu, Patricia Obeid, Donald K. Martin, Ali Bouamrani, Sophie Gerbaud, Benoit Gilquin, Thibaud Moufle-Milot, Adrien Mombrun, Cristina Belda Marin, Axel Tollance, Biomicrotechnologie et génomique fonctionnelle (BIOMICS), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Génie Enzymatique, Membrane Biomimétique et Assemblages Supramoléculaires (GEMBAS), Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Invasion mechanisms in angiogenesis and cancer (IMAC), Biologie du Cancer et de l'Infection (BCI ), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Gestes Medico-chirurgicaux Assistés par Ordinateur (TIMC-IMAG-GMCAO), Clinatec - Centre de recherche biomédicale Edmond J.Safra (SCLIN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])

Subjects
Exocrine gland, 3D scaffolds, Biomedical Engineering, Nanotechnology, polyelectrolytes, 03 medical and health sciences, 0302 clinical medicine, Prostate, Genetics, medicine, Liquid biopsy, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, liquid biopsy, Polyelectrolyte, secretome, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Molecular Medicine, organ-on-chip, TP248.13-248.65, Food Science, Biotechnology
Abstract

We report our approach to creating a microfluidic chip (namely UroLOC) that mimics the acinar/tubular structure and the luminal microenvironment of exocrine glands. The chip utilises a nanostructured membrane that is designed to provide a 3-dimensional supporting scaffold for the growth of exocrine acinus epithelial cells. The nanostructured membrane was produced using layer-by-layer assembly of polyelectrolytes, and formed into 3-dimensional hemispherical cavities and “finger-like” structures in order to mimic the natural architecture of acini found in exocrine glands. We utilised normal (PNT2) and cancerous (PC3, LNCaP) prostate epithelial cells to demonstrate the proof-of-concept of using MALDI (Matrix Assisted Laser Desorption Ionisation) profiling of secretions collected after 48 hours of cell growth, with no concentration or purification steps and without any a priori on the knowledge of targeted proteins. This MALDI profiling analysis of the crude supernatants from 3 different cell lines (PNT2, PC3 and LNCaP) demonstrated the capacity of the MALDI profiling approach to discriminate between the different secretome signatures. The UroLOC concept and secretome profiling that we describe opens new opportunities in terms of liquid-biopsy based diagnosis, particularly for the early stages of carcinogenesis.

Published
2018

8. La biopile enzymatique à glucose/oxygène Quelques nuances de Grays

Author

Donald K. Martin, Sarra El Ichi-Ribault, Laurent Cortella, Jean-Pierre Alcaraz, Catherine Guimier-Pingault, Abdelkader Zebda, Philippe Cinquin, Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), ARC-Nucleart CEA Grenoble (NUCLEART), Laboratoire d'Innovation pour les Technologies des Energies Nouvelles et les nanomatériaux (LITEN), Institut National de L'Energie Solaire (INES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS)-Institut National de L'Energie Solaire (INES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS), and Centre Hospitalier Universitaire [Grenoble] (CHU)

Subjects
0301 basic medicine, Laccase, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, biology, Chemistry, Oxygen metabolism, Implant Infection, 02 engineering and technology, General Medicine, 021001 nanoscience & nanotechnology, General Biochemistry, Genetics and Molecular Biology, Anode, 03 medical and health sciences, 030104 developmental biology, biology.protein, Glucose oxidase, 0210 nano-technology, Gamma irradiation, Biomedical engineering
Abstract

International audience; > Une biopile enzymatique implantable est un dispositif produisant du courant uniquement à partir du glucose et de l'oxygène naturellement et constamment présents dans l'organisme. Les enzymes présentes aux électrodes oxydent le glucose et réduisent l'oxygène, ce qui génère un flux d'électrons dans un circuit électrique associé. Cette microcentrale électrique, qui peut fournir une tension de presque un volt, pourra alimen-ter sur une longue période la future génération de robots implantables. Tout dispositif médical implantable doit être stérilisé avant son implan-tation. Comme le principe actif de cette biopile est composé d'enzymes, la question de la stérilisation de ce nouveau type de dispositif s'est posée. < pacemaker [2]. Notre équipe a récemment implanté chez le rat une bioélectrode modifiée qui a conservé son activité après une durée de 167 jours [3]. Utilisation de l'ionisation pour la stérilisation du dispositif L'implantation d'un dispositif médical électronique alimenté par une biopile pose le problème de sa stérilisation. Pour les applications contenant du matériel biologique, ce qui est le cas de la biopile enzy-matique, l'autoclavage est proscrit car les enzymes sont dénaturées par la chaleur. Deux types de stérilisation à froid restent alors pos-sibles : l'une utilisant le peroxyde d'hydrogène (H 2 O 2) sous vide d'air, l'autre les radiations ionisantes. La stérilisation au H 2 O 2 est un procédé couramment utilisé dans les hôpi-taux pour les dispositifs médicaux implantables. Malheureusement, la structure des bioélectrodes est fortement dégradée sous l'effet du vide partiel. De plus, les enzymes sont presque totalement inactivées par la dose d'agent oxydant. Nous nous sommes donc intéressés à la stérilisa-tion via les rayonnements ionisants (RI). L'action des RI sur la cellule a été théorisée il y a un siècle grâce aux travaux de Marie Curie [4] et s'utilise couramment lors des traitements en radiothérapie. La réponse cellulaire aux RI permet d'appréhender les mécanismes de radio-sensibilité [5] (➜) et d'optimiser les traitements (radiothérapie FLASH [6]) (➜). Qu'est-ce qu'une biopile implantable ? La biopile enzymatique implantable que nous dévelop-pons permet de générer de l'énergie électrique grâce à des enzymes immobilisées sur des électrodes, à par-tir de glucose et d'oxygène, substrats présents dans l'organisme et potentiellement inépuisables. Tant que les enzymes fonctionnent, et que les substrats diffusent librement, il est possible de fournir du courant de façon quasi illimitée à des dispositifs médicaux électro-niques pouvant suppléer des fonctions défaillantes chez l'homme. Le principe de fonctionnement est illustré par la Figure 1. Le glucose est oxydé à la bioanode par la glucose oxydase qui fournit des électrons au circuit. Après passage dans le circuit électrique, ces électrons sont récupérés au niveau de la biocathode. À ce niveau, la laccase 1 sert à réduire le dioxygène en eau. Ces bioélectrodes ont la forme d'une pastille contenant les enzymes immobilisées dans un réseau tridimensionnel de matériau nanostructuré. La preuve de concept in vivo a été réalisée par Cinquin et al. en 2010 [1]. Cette biopile fournit actuellement suffisamment d'éner-gie (40 μW) pour alimenter des dispositifs tels qu'un 1 Oxydase avec du cuivre comme cofacteur. (➜) Voir la Synthèse de N. Foray et al., m/s n° 4, avril 2013, page 397 (➜) Voir la Nouvelle de V. Favaudon et al., m/s n° 2, février 2015, page 121 Alcaraz_DossierTechnique.indd 771 Alcaraz_DossierTechnique.indd 771

Published
2016

9. Identification of three tomato flower and fruit MADS-box proteins with a putative histone deacetylase binding domain

Author

Marcel Kuntz, Joël Gaffé, Claudie Lemercier, Jean-Pierre Alcaraz, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Biochimie et biophysique des systèmes intégrés (BBSI), Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF), Plastes et Différenciation Cellulaire (PDC), Laboratoire de physiologie cellulaire végétale (LPCV), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
0106 biological sciences, MESH: Introns, MESH: Sequence Homology, Amino Acid, Sequence Homology, MESH: Amino Acid Sequence, MESH: Base Sequence, MESH: RNA, Plant, 01 natural sciences, Solanum lycopersicum, Genes, Reporter, MESH: Lycopersicon esculentum, MESH: Genes, Plant, MADS-box, Plant Proteins, Genetics, 0303 health sciences, Histone deacetylase 5, MESH: Plant Proteins, Histone deacetylase 2, Exons, General Medicine, Cell biology, DNA-Binding Proteins, Amino Acid, RNA, Plant, Histone deacetylase binding, MESH: DNA Primers, DNA, Plant, Molecular Sequence Data, MESH: Sequence Alignment, MADS Domain Proteins, Flowers, Biology, Genes, Plant, Histone Deacetylases, MESH: MADS Domain Proteins, 03 medical and health sciences, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Lycopersicon esculentum, MESH: DNA, Plant, Reporter, Transcription factor, DNA Primers, 030304 developmental biology, Binding Sites, MESH: Molecular Sequence Data, Sequence Homology, Amino Acid, Base Sequence, HDAC11, HDAC10, MESH: Genes, Reporter, DNA, Plant, MESH: Flowers, HDAC4, Introns, MESH: Histone Deacetylases, Genes, MESH: Binding Sites, RNA, MESH: Exons, Sequence Alignment, MESH: DNA-Binding Proteins, 010606 plant biology & botany
Abstract

International audience; MADS-box transcription factors play crucial roles in organ and cell differentiation in organisms ranging from yeast to humans. Most of the work on plant MADS-box proteins focused on their roles in floral development whereas less information is available on their function in fruit maturation. We cloned three distinct tomato cDNAs using a RT-PCR approach, encoding LeMADS1, LeMADS5 and LeMADS6 factors and whose mRNAs mostly accumulate in tomato flowers and fruits. Phylogeny analysis indicates that LeMADS1, 5 and 6 belong to the MEF2-like family. When transiently expressed in tobacco leaves or in human cells, LeMADS1, 5 and 6 are targeted to the cell nucleus. As the endogenous target genes of these putative transcription factors are unknown, the transcriptional activity of these proteins was characterized in a heterologous system and we showed that, when fused to a Gal4-DNA-binding domain, they repress the transcription of heterologous reporter genes. Since histone deacetylases control MEF2 transcriptional activity and since a putative histone deacetylase binding site was present in LeMADS1, 5 and 6, we tested the potential interaction between these factors and HDAC5 deacetylase. Surprisingly, in this heterologous system, LeMADS1, 5 and 6 interacted with HDAC5 N-terminal region. Our data suggest that, like mammalian MEF2A, plant MADS-box transcriptional activity might be regulated by enzymes controlling chromatin acetylation.

Published
2011

10. Plant lipid-associated fibrillin proteins condition jasmonate production under photosynthetic stress

Author

Abir Youssef, Joël Gaffé, Yec’han Laizet, Maryse A. Block, Dominique Pontier, Jean-Pierre Alcaraz, Eric Maréchal, Tony R. Larson, Marcel Kuntz, Plastes et Différenciation Cellulaire (PDC), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de physiologie cellulaire végétale (LPCV), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), CNAP, Department of Biology, University of York [York, UK], Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
0106 biological sciences, plastoglobule, Photoinhibition, Arabidopsis thaliana, Plastoglobule, thylakoid, Arabidopsis, plant, Plant Science, 01 natural sciences, Anthocyanins, chemistry.chemical_compound, Jasmonate, Photosynthesis, plastid, 0303 health sciences, Jasmonic acid, Microfilament Proteins, food and beverages, Chloroplast, Biochemistry, Thylakoid, RNA Interference, jasmonic acid biosynthesis, triacylglycerol, photooxidative stress, fibrillin, Cyclopentanes, Biology, lipid associated protein, Fibrillins, 03 medical and health sciences, chloroplast, Microscopy, Electron, Transmission, Stress, Physiological, Genetics, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Oxylipins, Plastid, 030304 developmental biology, Arabidopsis Proteins, fungi, photosystem II, Photosystem II Protein Complex, Cell Biology, biology.organism_classification, chemistry, photosynthesis acclimation, 010606 plant biology & botany
Abstract

International audience; Summary The role of a subfamily of lipid globule-associated proteins, referred to as plant fibrillins (FIB1a, -1b, -2), was determined using a RNA interference (RNAi) strategy. We show that Arabidopsis plants with reduced levels of these plastid structural proteins are impaired in long-term acclimation to environmental constraint, namely photooxidative stress imposed by high light combined with cold. As a result, their photosynthetic apparatus is inefficiently protected. This leads to the prevalence of an abnormal granal and stromal membrane arrangement, as well as higher photosystem II photoinhibition under stress. The visible phenotype of FIB1-2 RNAi lines also includes retarded shoot growth and a deficit in anthocyanin accumulation under stress. All examined phenotypic effects of lower FIB levels are abolished by jasmonate (JA) treatment. An atypical expression pattern of several JA-induced genes was observed in RNAi plants. A JA-deficient mutant was found to share similar stress phenotypic characteristics with FIB RNAi plants. We conclude a new physiological role for JA, namely acclimation of chloroplasts, and that light/cold stress-related JA biosynthesis is conditioned by the accumulation of plastoglobule-associated FIB1-2 proteins. Consistent correlative data suggest that this FIB effect is mediated by plastoglobule (and triacylglycerol) accumulation as the potential site for initiating the chloroplast stress-related JA biosynthesis.

Published
2010

11. Fibrillin influence on plastid ultrastructure and pigment content in tomato fruit

Author

Joël Gaffé, Paul D. Fraser, Andrew J. Simkin, Peter M. Bramley, Jean-Pierre Carde, Marcel Kuntz, Jean-Pierre Alcaraz, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Gestes Medico-chirurgicaux Assistés par Ordinateur (TIMC-IMAG-GMCAO), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Plastes et différenciation cellulaire (PDC), European commission Agriculture and Fisheries program for the mobility of researchers (Contract No. FAIR-98-5002), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Laboratoire de physiologie cellulaire végétale (LPCV), and Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, Plastoglobule, macromolecular substances, Plant Science, MESH: Capsicum, Horticulture, Biology, Fibrillins, Thylakoids, Chloroplast, 01 natural sciences, Biochemistry, MESH: Microfilament Proteins, 03 medical and health sciences, MESH: Thylakoids, Solanum lycopersicum, Membrane pore, MESH: Lycopersicon esculentum, Chromoplast, Plastids, Lycopersicon esculentum, Thylakoid integrity, Plastid, Molecular Biology, Carotenoid, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], Microfilament Proteins, food and beverages, Ripening, MESH: Plastids, General Medicine, Plants, Genetically Modified, Fruit ripening, Carotenoids, MESH: Plants, Genetically Modified, chemistry, Fruit, Thylakoid, MESH: Carotenoids, Capsicum, MESH: Fruit, Fibrillin, 010606 plant biology & botany
Abstract

International audience; The protein termed fibrillin is involved in the formation of lipoprotein structures, such as plastoglobules and fibrils in certain chromoplast types, which have been implicated in the over-production of pigments due to a sink effect. In order to examine its effect in differentiating chromoplasts of a non-fibrillar type, the pepper fibrillin gene was expressed in tomato fruit. Both the transcript and protein were found to accumulate during tomato fruit ripening from an early mature green stage. However, formation of carotenoid deposition structures in tomato chromoplasts, such as fibrils, was not observed. Nevertheless, a two-fold increase in carotenoid content and associated carotenoid derived flavour volatiles (6-methyl-5-hepten-2-one, geranylacetone, beta-ionone and beta-cyclocitral) was observed. An unexpected phenotypic observation in the transgenic fruit was the delayed loss of thylakoids in differentiating chromoplasts, leading to the transient formation of plastids exhibiting a typical chromoplastic zone adjacent to a protected chloroplastic zone with preserved thylakoids. An in vitro assay has been developed to monitor fibrillin activity on thylakoids: data were obtained suggesting a membrane protection role for fibrillin, more specifically against moderate uncoupling effects.

Published
2007

12. Intraplastidial trafficking of a phage-type RNA polymerase is mediated by a thylacoïd RING-H2 protein

Author

Silva Lerbs-Mache, Thierry Lagrange, Mohamed-Ali Hakimi, Pankaj Jaiswal, Florence Courtois, Emilie Demarsy, Laurence Maréchal-Drouard, Jacinthe Azevedo, Jean-Pierre Alcaraz, Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Plastes et différenciation cellulaire (PDC), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), Center for Integrative Genomics - Institute of Bioinformatics, Génopode (CIG), Swiss Institute of Bioinformatics [Lausanne] (SIB), Université de Lausanne (UNIL)-Université de Lausanne (UNIL), Laboratoire Génome et développement des plantes (LGDP), Université de Perpignan Via Domitia (UPVD)-Centre National de la Recherche Scientifique (CNRS), Department of Plant Breeding and Genetics, Cornell University [New York], and Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, Light, Arabidopsis, MESH: Amino Acid Sequence, Thylakoids, 01 natural sciences, chemistry.chemical_compound, MESH: Saccharomyces cerevisiae Proteins, MESH: Thylakoids, Gene Expression Regulation, Plant, Transcription (biology), RNA, Ribosomal, 16S, RNA polymerase, MESH: Arabidopsis, Bacteriophages, Promoter Regions, Genetic, MESH: Organ Specificity, 0303 health sciences, Multidisciplinary, food and beverages, MESH: Transcription Factors, DNA-Directed RNA Polymerases, Biological Sciences, Transport protein, Cell biology, MESH: RNA, Ribosomal, 16S, DNA-Binding Proteins, Chloroplast, MESH: Promoter Regions (Genetics), MESH: Intracellular Membranes, Protein Transport, Transmembrane domain, Organ Specificity, Thylakoid, Protein Binding, MESH: Protein Transport, Saccharomyces cerevisiae Proteins, Molecular Sequence Data, MESH: Arabidopsis Proteins, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, MESH: Two-Hybrid System Techniques, DNA-binding protein, 03 medical and health sciences, Two-Hybrid System Techniques, MESH: Gene Library, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Bacteriophages, Amino Acid Sequence, RNA, Messenger, MESH: Gene Expression Regulation, Plant, Plastid, MESH: RNA, Messenger, Gene Library, 030304 developmental biology, MESH: Molecular Sequence Data, Arabidopsis Proteins, Intracellular Membranes, Molecular biology, MESH: Light, MESH: DNA-Directed RNA Polymerases, chemistry, Transcription Factors, 010606 plant biology & botany
Abstract

The plastid genome of dicotyledonous plants is transcribed by three different RNA polymerases; an eubacterial-type enzyme, PEP; and two phage-type enzymes, RPOTp and RPOTmp. RPOTp plays an important role in chloroplast transcription, biogenesis, and mesophyll cell proliferation. RPOTmp fulfills a specific function in the transcription of the rrn operon in proplasts/amyloplasts during seed imbibition/germination and a more general function in chloroplasts during later developmental stages. In chloroplasts, RPOTmp is tightly associated with thylakoid membranes indicating that functional switching of RPOTmp is connected to thylakoid association. By using the yeast two-hybrid system, we have identified two proteins that interact with RPOTmp. The two proteins are very similar, both characterized by three N-terminal transmembrane domains and a C-terminal RING domain. We show that at least one of these proteins is an intrinsic thylakoid membrane protein that fixes RPOTmp on the stromal side of the thylakoid membrane, probably via the RING domain. A model is presented in which light by triggering the synthesis of the RING protein determines membrane association and functional switching of RPOTmp.

Published
2008

13. Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria

Author

Nadège Philippe, Dominique Schneider, Evelyne Coursange, Jean-Pierre Alcaraz, Johannes Geiselmann, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Plastes et différenciation cellulaire (PDC), and Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
MESH: DNA Primers, Biology, Bacterial genetics, 03 medical and health sciences, Plasmid, Amp resistance, MESH: Plasmids, Escherichia coli, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Insertion sequence, Molecular Biology, Gene, Alleles, 030304 developmental biology, DNA Primers, Genetics, 0303 health sciences, 030306 microbiology, MESH: Escherichia coli, Point mutation, MESH: Alleles, Genes, Transgenic, Suicide, Suicide gene, Transformation (genetics), MESH: Genes, Transgenic, Suicide, MESH: DNA Transposable Elements, DNA Transposable Elements, Transformation, Bacterial, MESH: Transformation, Bacterial, Plasmids
Abstract

Allelic exchange experiments allow investigation of the functions of many unknown genes identified during the sequencing of entire genomes. Isogenic strains differing by only specific mutations can be constructed. Among other tools, suicide plasmids are widely used for this task. They present many advantages because they leave no scars on the chromosome, and therefore allow combining several mutations in the same genetic background. While using the previously described pCVD442 suicide plasmid [Infect. Immun. 59 (1991) 4310], we found untargeted recombination events due to the presence of an IS1 element on this plasmid. The plasmid was therefore improved by removal of the IS1 element. We also replaced the bla gene of pCVD442, conferring ampicillin resistance, by the cat gene conferring chloramphenicol resistance, leading to the new suicide plasmid pDS132. The plasmid was entirely sequenced. We demonstrate that this new vector can be easily used to introduce various types of mutations into different genetics backgrounds: removal of IS elements, introduction of point mutations or deletions. It can be introduced into bacterial strains by either transformation or conjugation.

Published
2004

14. Tackling the Concept of Symbiotic Implantable Medical Devices with Nanobiotechnologies

Author

Jean-Pierre Alcaraz, Donald K. Martin, Philippe Cinquin, Systèmes Nanobiotechnologiques et Biomimétiques (TIMC-IMAG-SyNaBi), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Gestes Medico-chirurgicaux Assistés par Ordinateur (TIMC-IMAG-GMCAO), and Alcaraz, Jean-Pierre

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Computer science, Biocompatible Materials, 02 engineering and technology, Regenerative Medicine, Applied Microbiology and Biotechnology, 03 medical and health sciences, Human–computer interaction, Humans, Nanotechnology, Implanted device, ComputingMilieux_MISCELLANEOUS, Prostheses and Implants, General Medicine, 021001 nanoscience & nanotechnology, Biocompatible material, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, 3. Good health, Encapsulation (networking), 030104 developmental biology, Equipment and Supplies, Molecular Medicine, Bioinspiration, 0210 nano-technology, Biotechnology
Abstract

This review takes an approach to implanted medical devices that considers whether the intention of the implanted device is to have any communication of energy or materials with the body. The first part describes some specific examples of three different classes of implants, analyzed with regards to the type of signal sent to cells. Through several examples, the authors describe that a one way signaling to the body leads to encapsulation or degradation. In most cases, those phenomena do not lead to major problems. However, encapsulation or degradation are critical for new kinds of medical devices capable of duplex communication, which are defined in this review as symbiotic devices. The concept the authors propose is that implanted medical devices that need to be symbiotic with the body also need to be designed with an intended duplex communication of energy and materials with the body. This extends the definition of a biocompatible system to one that requires stable exchange of materials between the implanted device and the body. Having this novel concept in mind will guide research in a new field between medical implant and regenerative medicine to create actual symbiotic devices.

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