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1. Genetically tailored magnetosomes used as MRI probe for molecular imaging of brain tumor

Author

Sandra Prévéral, Erwan Selingue, Sébastien Mériaux, Marianne Boucher, David Pignol, Christopher T. Lefèvre, Michel Pean, Géraldine Adryanczyk-Perrier, Françoise Geffroy, Nicolas Ginet, L. Bellanger, HYBIS, Laboratoire d'étude des transferts en hydrologie et environnement (LTHE), Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Grenoble (INPG)-Institut national des sciences de l'Univers (INSU - CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Observatoire des Sciences de l'Univers de Grenoble (OSUG), Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Grenoble (INPG)-Institut national des sciences de l'Univers (INSU - CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Observatoire des Sciences de l'Univers de Grenoble (OSUG), Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Department of Applied Geophysics, Technische Universität Berlin (TUB)-Technische Universität Berlin (TUB), Unité d'imagerie par résonance magnétique à très haut champ et de spectroscopie (UNIRS), Service NEUROSPIN (NEUROSPIN), 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)-Université Paris-Saclay-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)-Université Paris-Saclay, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-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), Microbiologie Environnementale et Moléculaire (MEM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Service de Pharmacologie et Immunoanalyse (SPI), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Laboratoire de chimie bactérienne (LCB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Observatoire des Sciences de l'Univers de Grenoble (OSUG ), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut national des sciences de l'Univers (INSU - CNRS)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Service de Pharmacologie et d'Immunoanalyse (SPI), Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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 Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-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)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0301 basic medicine, Materials science, media_common.quotation_subject, Magnetosome, Biophysics, Brain tumor, Contrast Media, Molecular Probe Techniques, Bioengineering, Single step, Nanoconjugates, 02 engineering and technology, Article, Biomaterials, Mice, 03 medical and health sciences, Nuclear magnetic resonance, Cell Line, Tumor, Biomarkers, Tumor, medicine, Animals, Tissue Distribution, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Internalization, media_common, medicine.diagnostic_test, Brain Neoplasms, RGD peptide, Magnetic resonance imaging, 021001 nanoscience & nanotechnology, medicine.disease, Magnetic Resonance Imaging, Molecular Imaging, Genetic Enhancement, 030104 developmental biology, Mechanics of Materials, Molecular Probes, Ceramics and Composites, Magnetosomes, Lipid vesicle, Molecular imaging, 0210 nano-technology, Oligopeptides
Abstract

International audience; We investigate here the potential of single step production of genetically engineeredmagnetosomes, bacterial biogenic iron-oxide nanoparticles embedded in a lipid vesicle, as a new tailorable magnetic resonance molecular imaging probe. We demonstrate in vitro the specific binding and the significant internalization into U87 cells of magnetosomes decorated with RGD peptide. After injection at the tail vein of glioblastoma-bearing mice, we evidence in the first 2 h the rapid accumulation of both unlabeled and functionalized magnetosomes inside the tumor by Enhanced Permeability and Retention effects. 24 h after the injection, a specific enhancement of the tumor contrast is observed on MR images only for RGD-labeled magnetosomes. Post mortem acquisition of histological data confirms MRI results with more magnetosomes found into the tumor treated with functionalized magnetosomes. This work establishes the first proof-of-concept of a successful bio-integrated production of molecular imaging probe for MRI.

Published
2017

2. Evidencing 98 secondary metabolites of Penicillium verrucosum using substrate isotopic labeling and high-resolution mass spectrometry

Author

Isabelle P. Oswald, Michel Pean, Olivier Puel, Thaïs Hautbergue, Emilien L. Jamin, Souria Tadrist, Marcel Delaforge, Lauriane Meneghetti, Laurent Debrauwer, ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), Axiom Platform, MetaToul-MetaboHUB, National Infrastructure of Metabolomics and Fluxomics, Groupe de Recherches Appliquées en Phytotechnologie (GRAP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Microbiologie Environnementale et Moléculaire (MEM), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-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)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-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), Système membranaires, photobiologie, stress et détoxication (SMPSD), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), French National Infrastructureof Metabolomics and Fluxomics (MetaboHUB-ANR-11-INBS-0010), ANR-15-CE21-0010,Newmyco,Metabolome de deux contaminants fongiques du blé d'importance majeure: identification de nouveaux métabolites toxiques(2015), Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Ecole d'Ingénieurs de Purpan (INP - PURPAN), Université de Toulouse (UT)-Université de Toulouse (UT), Biosynthèse & Toxicité des Mycotoxines (ToxAlim-BioToMyc), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), MetaToul AXIOM (E20), Université de Toulouse (UT)-Université de Toulouse (UT)-MetaboHUB-MetaToul, MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-MetaboHUB-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Puel, Olivier, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), 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)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Analyse de Xénobiotiques, Identification, Métabolisme (E20 Metatoul-AXIOM), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-MetaToul-MetaboHUB, Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-MetaToul-MetaboHUB, Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0301 basic medicine, penicillium verrucosum, [SDV]Life Sciences [q-bio], Clinical Biochemistry, mass spectrum analysis, toxicologie alimentaire, stable isotope labeling, Fungus, secondary metabolite, Secondary metabolite, Mass spectrometry, Biochemistry, Analytical Chemistry, Isotopic labeling, 03 medical and health sciences, Metabolomics, spectrométrie de masse, medicine, Penicillium verrucosum, Metabolome, Chromatography, High Pressure Liquid, mass spectrometry, molecular networking, Chromatography, biology, Chemistry, public health, fungus, Penicillium, Cell Biology, General Medicine, biology.organism_classification, 030104 developmental biology, métabolite secondaire, santé publique, Isotope Labeling, secondary plant products, medicine.drug
Abstract

Industrial applications of fungal compounds, coupled with the emergence of fungal threats to natural ecosystems and public health, have increased interest in filamentous fungi. Among all pathogenic fungi, Penicillium verrucosum is one of the most common mold-infecting stored cereals in temperate regions. However, it is estimated that 80% of fungal secondary metabolites remain unknown. To detect new P. verrucosum compounds, an untargeted metabolomic approach was applied to fungus grown on wheat grains labeled with stable isotopes: (i) natural grains (99% 12C); (ii) grains enriched with 97% of 13C; and (iii) grains enriched with 53% of 13C and 97% of 15N. Analyses performed by high-performance liquid chromatography coupled with high-resolution mass spectrometry (HPLC-HRMS) enabled the specific detection of fungal metabolites, and the unambiguous characterization of their chemical formulas. In this way, 98 secondary metabolites were detected and their chemical formulas were determined. Of these, only 18 identifications could be made based on databases, the literature and mass spectrometry fragmentation experiments, with the result that 80 were totally unknown. Molecular networks were generated to analyze these results, leading to the characterization by MSn experiments of a new fungisporin produced by P. verrucosum. More generally, this article provides precise mass spectrometric data about all these compounds for further studies of the Penicillium metabolome.

Published
2017

3. GOLLUM [FeFe]-hydrogenase-like proteins are essential for plant development in normoxic conditions and modulate energy metabolism

Author

Guillaume Tcherkez, Pascal Ratet, Jean-Malo Couzigou, Françoise Gilard, Stéphanie Pateyron, Aurore Lenglet, Viviane Cosson, Michel Pean, Lysiane Brocard, Samuel Mondy, Sandra Pelletier, and Marije Scholte

Subjects
0106 biological sciences, chemistry.chemical_classification, 0303 health sciences, Mutation, Hydrogenase, Physiology, Protein subunit, Mutant, Plant Science, Biology, biology.organism_classification, medicine.disease_cause, 01 natural sciences, 03 medical and health sciences, Enzyme, chemistry, Biochemistry, Arabidopsis, medicine, Arabidopsis thaliana, Gene, 030304 developmental biology, 010606 plant biology & botany
Abstract

[FeFe]-hydrogenase-like genes encode [Fe 4 S 4 ]-containing proteins that are ubiquitous in eukaryotic cells. In humans, iron-only hydrogenase-like protein 1 (IOP1) represses hypoxia inducible factor-1α subunit (HIF1-α) at normal atmospheric partial O 2 pressure (normoxia, 21 kPa O 2 ). In yeasts, the narl mutant cannot grow at 21 kPa O 2 . but can develop at a lower O 2 pressure (2 kPa O 2 ). We show here that plant [FeFe]-hydrogenase-like GOLLUM genes are essential for plant development and cell cycle progression. The mutant phenotypes of these plants are seen in normoxic conditions, but not under conditions of mild hypoxia (5 kPa O 2 ). Transcriptomic and metabolomic experiments showed that the mutation enhances the expression of some hypoxia-induced genes under normal atmospheric O 2 conditions and changes the cellular content of metabolites related to energy metabolism. In conclusion, [FeFe]-hydrogenase-like proteins play a central role in eukaryotes including the adaptation of plants to the ambient O 2 partial pressure.

Published
2013

4. Contribution of uniformly 13C-enriched sterigmatocystin to the study of its pulmonary metabolism

Author

Odile Cabaret, Michel Pean, Françoise Botterel, Stéphane Bretagne, Marcel Delaforge, and Olivier Puel

Subjects
chemistry.chemical_classification, 0303 health sciences, Aflatoxin, biology, Cytochrome, fungi, Organic Chemistry, Cytochrome P450, Metabolism, Glutathione, Analytical Chemistry, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Enzyme, chemistry, Biochemistry, 030220 oncology & carcinogenesis, biology.protein, Respiratory epithelium, Spectroscopy, 030304 developmental biology, Sterigmatocystin
Abstract

Mycotoxins are secondary metabolites of filamentous fungi which can cause a wide range of systemic effects. Human health effects of inhaled mycotoxins remain poorly documented, despite the large amounts present, associated with air-borne particles. Among these mycotoxins, sterigmatocystin is one of the most prevalent. Because its chemical structure is close to that of the aflatoxins, we studied its metabolism and its cellular consequences when in contact with the airway epithelium, using the mass spectral signature from the 10% 13C uniformly enriched sterigmatocystin. The metabolism was studied in vitro, using recombinant cytochrome P450s enzymes, and in porcine tracheal epithelial cell (PTEC) primary cultures at an air-liquid interface. The metabolites were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry detection. Expressed enzymes and PTECs were exposed to uniformly 13 C-enriched sterigmatocystin to confirm the relationship between sterigmatocystin and its metabolites because this isotopic cluster shape is conserved for all metabolites and their product ions. Incubation of sterigmatocystin with recombinant cytochrome P450 1A1 led to the formation of three metabolites identified as monohydroxysterigmatocystin, dihydroxysterigmatocystin and one glutathione adduct, the latter after the formation of a transient intermediate. In the PTEC cultures, sterigmatocystin metabolism resulted in a glucuro-conjugate. Two other products were detected, a sulfo-conjugate and a glucuro-conjugate of hydroxysterigmatocystin upon cytochrome P450 1A1 induction. This is the first study to report sterigmatocystin metabolism in airway epithelium, and it suggests that, contrary to the aflatoxins, sterigmatocystin is mainly detoxified into its conjugates and is unable to produce significant amounts of reactive metabolites in respiratory cells, at least in pigs. Copyright © 2011 John Wiley & Sons, Ltd.

Published
2011

5. NMR study of cellulose and wheat straw degradation by Ruminococcus albus 20

Author

Mária Matulová, Evelyne Forano, Régis Nouaille, Anne-Marie Delort, Michel Pean, and Peter Capek

Subjects
Arabinose, 0303 health sciences, animal structures, Fibrobacter succinogenes, 030306 microbiology, food and beverages, Cell Biology, Xylose, Straw, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, Hydrolysis, chemistry, Cellodextrin, Hemicellulose, Food science, Cellulose, Molecular Biology, 030304 developmental biology
Abstract

Cellulose and wheat straw degradation by Ruminococcus albus was monitored using NMR spectroscopy. In situ solid-state (13)C-cross-polarization magic angle spinning NMR was used to monitor the modification of the composition and structure of cellulose and (13)C-enriched wheat straw during the growth of the bacterium on these substrates. In cellulose, amorphous regions were not preferentially degraded relative to crystalline areas by R. albus. Cellulose and hemicelluloses were also degraded at the same rate in wheat straw. Liquid state two-dimensional NMR experiments were used to analyse in detail the sugars released in the culture medium, and the integration of NMR signals enabled their quantification at various times of culture. The results showed glucose and cellodextrin accumulation in the medium of cellulose cultures; the cellodextrins were mainly cellotriose and accumulated to up to 2 mm after 4 days. In the wheat straw cultures, xylose was the main soluble sugar detected (1.4 mm); arabinose and glucose were also found, together with some oligosaccharides liberated from hemicellulose hydrolysis, but to a much lesser extent. No cellodextrins were detected. The results indicate that this strain of R. albus is unable to use glucose, xylose and arabinose for growth, but utilizes efficiently xylooligosaccharides. R. albus 20 appears to be less efficient than Fibrobacter succinogenes S85 for the degradation of wheat straw.

Published
2008

6. Strong decrease in lignin content without significant alteration of plant development is induced by simultaneous down-regulation of cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) in tobacco plants

Author

Saïda Danoun, Michel Pean, and Alain Michel Boudet, Jacqueline Grima-Pettenati, Matthieu Chabannes, John Ralph, Claire Halpin, Catherine Lapierre, Jane M. Marita, and Abdellah Barakate

Subjects
Transgene, Cinnamyl-alcohol dehydrogenase, Nicotiana tabacum, fungi, Wild type, Cell Biology, Plant Science, Biology, biology.organism_classification, chemistry.chemical_compound, Biochemistry, chemistry, Genetics, Cinnamoyl-CoA reductase, Lignin, Ectopic expression, Monolignol
Abstract

Different transgenic tobacco lines down-regulated for either one or two enzymes of the monolignol pathway were compared for their lignin content and composition, and developmental patterns. The comparison concerned CCR and CAD down-regulated lines (homozygous or heterozygous for the transgene) and the hybrids resulting from the crossing of transgenic lines individually altered for CCR or CAD activities. Surprisingly, the crosses containing only one allele of each antisense transgene, exhibit a dramatic reduction of lignin content similar to the CCR down-regulated parent but, in contrast to this transgenic line, display a normal phenotype and only slight alterations of the shape of the vessels. Qualitatively the lignin of the double transformant displays characteristics more like the wild type control than either of the other transgenics. In the transgenics with a low lignin content, the transformations induced other biochemical changes involving polysaccharides, phenolic components of the cell wall and also soluble phenolics. These results show that the ectopic expression of a specific transgene may have a different impact depending on the genetic background and suggest that the two transgenes present in the crosses may operate synergistically to reduce the lignin content. In addition, these data confirm that plants with a severe reduction in lignin content may undergo normal development at least in controlled conditions.

Published
2001

7. NMR characterization of altered lignins extracted from tobacco plants down-regulated for lignification enzymes cinnamylalcohol dehydrogenase and cinnamoyl-CoA reductase

Author

Alain M. Boudet, Nabila Yahiaoui, Ronald D. Hatfield, John Ralph, Joël Piquemal, Catherine Lapierre, and Michel Pean

Subjects
Multidisciplinary, Sinapaldehyde, biology, Nicotiana tabacum, fungi, food and beverages, Reductase, biology.organism_classification, Cell wall, chemistry.chemical_compound, chemistry, Biochemistry, biology.protein, Cinnamoyl-CoA reductase, Lignin, Coniferyl alcohol, Alcohol dehydrogenase
Abstract

Homologous antisense constructs were used to down-regulate tobacco cinnamyl-alcohol dehydrogenase (CAD; EC 1.1.1.195 ) and cinnamoyl-CoA reductase (CCR; EC 1.2.1.44 ) activities in the lignin monomer biosynthetic pathway. CCR converts activated cinnamic acids (hydroxycinnamoyl–SCoAs) to cinnamaldehydes; cinnamaldehydes are then reduced to cinnamyl alcohols by CAD. The transformations caused the incorporation of nontraditional components into the extractable tobacco lignins, as evidenced by NMR. Isolated lignin of antisense-CAD tobacco contained fewer coniferyl and sinapyl alcohol-derived units that were compensated for by elevated levels of benzaldehydes and cinnamaldehydes. Products from radical coupling of cinnamaldehydes, particularly sinapaldehyde, which were barely discernible in normal tobacco, were major components of the antisense-CAD tobacco lignin. Lignin content was reduced in antisense-CCR tobacco, which displayed a markedly reduced vigor. That lignin contained fewer coniferyl alcohol-derived units and significant levels of tyramine ferulate. Tyramine ferulate is a sink for the anticipated build-up of feruloyl–SCoA, and may be up-regulated in response to a deficit of coniferyl alcohol. Although it is not yet clear whether the modified lignins are true structural components of the cell wall, the findings provide further indications of the metabolic plasticity of plant lignification. An ability to produce lignin from alternative monomers would open new avenues for manipulation of lignin by genetic biotechnologies.

Published
1998

8. Stimulation of different functional groups of bacteria by various plant residues as a driver of soil priming effect

Author

Séverine Boiry, Lionel Ranjard, Christophe Mougel, Sébastien Terrat, Noémie Pascault, Pierre-Alain Maron, Jean Lévêque, Richard Christen, Michel Pean, Aurore Kaisermann, Olivier Mathieu, Dipankar Bachar, Catherine Hénault, Sébastien Fontaine, Philippe Lemanceau, Agroécologie [Dijon], Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Institut de signalisation, biologie du développement et cancer (ISBDC), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Biogéosciences [UMR 6282] (BGS), Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), Groupe de Recherches Appliquées en Phytotechnologie (GRAP), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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), Unité de recherche sur l'Ecosystème Prairial (UREP), Institut National de la Recherche Agronomique (INRA), Study financially supported by the Agence de l'Environnement et de la Maîtrise de l'Energie (ADEME) and the Burgundy region., European Project: 264465,EC:FP7:ENV,FP7-ENV-2010,ECOFINDERS(2011), Université Nice Sophia Antipolis (... - 2019) (UNS), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Biogéosciences [UMR 6282] [Dijon] (BGS), Centre National de la Recherche Scientifique (CNRS)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), UR 0874 Unité de recherche sur l'Ecosystème Prairial, Institut National de la Recherche Agronomique (INRA)-Unité de recherche sur l'Ecosystème Prairial (UREP)-Ecologie des Forêts, Prairies et milieux Aquatiques (EFPA), Université Côte d'Azur (UCA), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche sur l'Ecosystème Prairial - UMR (UREP), and Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)

Subjects
Firmicutes, [SDE.MCG]Environmental Sciences/Global Changes, Stable-isotope probing, [SDV.SA.SDS]Life Sciences [q-bio]/Agricultural sciences/Soil study, Decomposer, soil, 03 medical and health sciences, [SDU.STU.GC]Sciences of the Universe [physics]/Earth Sciences/Geochemistry, soil organic matter, Botany, carbon cycle, Environmental Chemistry, Gemmatimonadetes, Organic matter, stable isotope probing, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 2. Zero hunger, chemistry.chemical_classification, 0303 health sciences, Ecology, biology, Soil organic matter, Verrucomicrobia, bacterial diversity, food and beverages, 04 agricultural and veterinary sciences, 15. Life on land, biology.organism_classification, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, pyrosequencing, Agronomy, chemistry, 040103 agronomy & agriculture, 0401 agriculture, forestry, and fisheries, Acidobacteria
Abstract

13 pages; International audience; The turnover of organic matter in soil depends on the activity of microbial decomposers. However, little is known about how modifications of the diversity of soil microbial communities induced by fresh organic matter (FOM) inputs can regulate carbon cycling. Here, we investigated the decomposition of two 13C labeled crop residues (wheat and alfalfa) and the dynamics of the genetic structure and taxonomic composition of the soil bacterial communities decomposing 13C labeled FOM and native unlabeled soil organic matter (SOM), respectively. It was achieved by combining the stable isotope probing method with molecular tools (DNA genotyping and pyrosequencing of 16S rDNA). Although a priming effect (PE) was always induced by residue addition, its intensity increased with the degradability of the plant residue. The input of both wheat and alfalfa residues induced a rapid dynamics of FOM-degrading communities, corresponding to the stimulation of bacterial phyla which have been previously described as copiotrophic organisms. However, the dynamics and the identity of the bacterial groups stimulated depended on the residue added, with Firmicutes dominating in the wheat treatment and Proteobacteria dominating in the alfalfa treatment after 3 days of incubation. In both treatments, SOMdegrading communities were dominated by Acidobacteria, Verrucomicrobia, and Gemmatimonadetes phyla which have been previously described as oligotrophic organisms. An early stimulation of SOM-degrading populations mainly belonging to Firmicutes and Bacteroidetes groups was observed in the alfalfa treatment whereas no change occurred in the wheat treatment. Our findings support the hypothesis that the succession of bacterial taxonomic groups occurring in SOM- and FOMdegrading communities during the degradation process may be an important driver of the PE, and consequently of carbon dynamics in soil.

Published
2013

9. In situ recordings of large gelatinous spheres from NE Atlantic, and the first genetic confirmation of egg mass of Illex coindetii (Vérany, 1839) (Cephalopoda, Mollusca)

Author

Halldis Ringvold, Morag Taite, A. Louise Allcock, Michael Vecchione, Michel Péan, Roberto Sandulli, Geir Johnsen, Arne Fjellheim, Snorre Bakke, Hanne Sannæs, Ann-Elin Wårøy Synnes, José Coronel, Martin Hansen, Peter G. Olejar, Geir Eliassen, Anita Eliassen, and Karl Klungland

Subjects
Medicine, Science
Abstract

Abstract In total, 90 gelatinous spheres, averaging one meter in diameter, have been recorded from ~ 1985 to 2019 from the NE Atlantic Ocean, including the Mediterranean Sea, using citizen science. More than 50% had a dark streak through center. They were recorded from the surface to ~ 60–70 m depth, mainly neutrally buoyant, in temperatures between 8 and 24°C. Lack of tissue samples has until now, prohibited confirmation of species. However, in 2019 scuba divers secured four tissue samples from the Norwegian coast. In the present study, DNA analysis using COI confirms species identity as the ommastrephid broadtail shortfin squid Illex coindetii (Vérany, 1839); these are the first confirmed records from the wild. Squid embryos at different stages were found in different egg masses: (1) recently fertilized eggs (stage ~ 3), (2) organogenesis (stages ~ 17–19 and ~ 23), and (3) developed embryo (stage ~ 30). Without tissue samples from each and every record for DNA corroboration we cannot be certain that all spherical egg masses are conspecific, or that the remaining 86 observed spheres belong to Illex coindetii. However, due to similar morphology and size of these spheres, relative to the four spheres with DNA analysis, we suspect that many of them were made by I. coindetii.

Published
2021
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10. Degradation of wheat straw by Fibrobacter succinogenes S85: a liquid and solid state Nuclear Magnetic resonance study

Author

Anne-Marie Delort, Régis Nouaille, Evelyne Forano, Mária Matulová, Michel Pean, Peter Capek, Synthèse et étude de systèmes à intêret biologique (SEESIB), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Dietary Fiber, Magnetic Resonance Spectroscopy, Rumen, Oligosaccharides, In Vitro Techniques, Polysaccharide, Applied Microbiology and Biotechnology, Lignin, 03 medical and health sciences, chemistry.chemical_compound, WHEAT STRAW, FIBROBACTER SUCCINOGENES, Polysaccharides, Animals, Cellulose, Acetylxylan esterase, MICROBIAL LIGNINOLYSIS, Triticum, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Fibrobacter succinogenes, Chromatography, Ecology, 030306 microbiology, food and beverages, Nuclear magnetic resonance spectroscopy, Straw, Culture Media, Kinetics, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, chemistry, Biochemistry, Fibrolytic bacterium, Biodegradation, Fibrobacter, Food Science, Biotechnology
Abstract

Wheat straw degradation by Fibrobacter succinogenes was monitored by nuclear magnetic resonance (NMR) spectroscopy and chemolytic methods to investigate the activity of an entire fibrolytic system on an intact complex substrate. In situ solid-state NMR with 13 C cross-polarization magic angle spinning was used to monitor the modification of the composition and structure of lignocellulosic fibers (of 13 C-enriched wheat straw) during the growth of bacteria on this substrate. There was no preferential degradation either of amorphous regions of cellulose versus crystalline regions or of cellulose versus hemicelluloses in wheat straw. This suggests either a simultaneous degradation of the amorphous and crystalline parts of cellulose and of cellulose and hemicelluloses by the enzymes or degradation at the surface at a molecular scale that cannot be detected by NMR. Liquid-state two-dimensional NMR experiments and chemolytic methods were used to analyze in detail the various sugars released into the culture medium. An integration of NMR signals enabled the quantification of oligosaccharides produced from wheat straw at various times of culture and showed the sequential activities of some of the fibrolytic enzymes of F. succinogenes S85 on wheat straw. In particular, acetylxylan esterase appeared to be more active than arabinofuranosidase, which was more active than α-glucuronidase. Finally, cellodextrins did not accumulate to a great extent in the culture medium.

Published
2005

11. Glucose appearance in the peripheral circulation and liver glucose output in men after a large 13C starch meal

Author

François Péronnet, Marion Korach-André, Xavier Leverve, Hubert Roth, D. Barnoud, Michel Pean, Laboratoire de bioénergétique fondamentale et appliquée (LBFA), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Nutrition Parentérale, CHU Grenoble-Hôpital Michallon, Groupe de Recherches Appliquées en Phytotechnologie (GRAP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Département de Kinésiologie, Université de Montréal (UdeM), Hamant, Sarah, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-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
MESH: Oxidation-Reduction, Blood Glucose, Male, 030309 nutrition & dietetics, Starch, medicine.medical_treatment, Medicine (miscellaneous), chemistry.chemical_compound, 0302 clinical medicine, Ingestion, Insulin, Urea, Food science, MESH: Biological Availability, 2. Zero hunger, 0303 health sciences, Meal, Carbon Isotopes, Nutrition and Dietetics, food and beverages, MESH: Oryza sativa, MESH: Glucose, Liver, Area Under Curve, Lipogenesis, Digestion, MESH: Calorimetry, Indirect, MESH: Digestion, Oxidation-Reduction, Adult, medicine.medical_specialty, MESH: Starch, MESH: Intestinal Absorption, Biological Availability, 030209 endocrinology & metabolism, MESH: Insulin, Excretion, 03 medical and health sciences, Internal medicine, medicine, Humans, MESH: Fatty Acids, Volatile, MESH: Urea, MESH: Humans, MESH: Carbon Isotopes, MESH: Adult, Calorimetry, Indirect, Oryza, Fatty Acids, Volatile, MESH: Male, Endocrinology, Glucose, chemistry, Intestinal Absorption, Glycemic Index, MESH: Blood Glucose, MESH: Glycemic Index, MESH: Area Under Curve, MESH: Liver
Abstract

International audience; BACKGROUND: Glucose absorption from starchy food has only been described with small amounts ingested ( approximately 20-75 g). OBJECTIVE: Our aim was to describe total plasma (Ra) and exogenous glucose (Ra(exo)) appearance, glucose release from the liver (HGP), and the metabolic response after ingestion of 5 g polished or parboiled rice/kg body mass. DESIGN: Gas exchange and urea excretion were monitored in 8 healthy subjects before (3.5 h) and after (8 h) ingestion of rice intrinsically labeled with (13)C; [6,6-(2)H(2)]glucose was infused for the measurement of Ra, Ra(exo), and HGP. RESULTS: Changes in plasma glucose, insulin, lactate, and free fatty acids and the increase in Ra(exo) and Ra ( approximately 200%) and the decrease in HGP ( approximately 90%) were not significantly different (P > 0.05) after ingestion of either rice. Glucose oxidation was not significantly different (111.6 +/- 8.2 compared with 89.0 +/- 11.3 g; P = 0.13), but fat oxidation was significantly lower (9.9 +/- 1.7 compared with 21.3 +/- 4.0 g; P < 0.05) after parboiled than after polished rice. The percentage of the glucose load that appeared in the circulation over 8 h was not significantly different after ingestion of polished (70.4 +/- 4.5%) or parboiled (63.8 +/- 2.0%) rice (P > 0.05). CONCLUSION: Although the starch in parboiled rice is less susceptible to digestion in vitro, exogenous glucose availability was not significantly different after ingestion of large amounts of polished or parboiled rice. Glucose absorption remains incomplete 8 h after ingestion of both types of rice.

Published
2004

12. Mass spectrometric determination of the inorganic carbon species assimilated by photoautotrophic cells of Euphorbia characias L

Author

F Rebeille, P Tapie, P Chagvardieff, Michel Pean, P Gans, and P Thibault

Subjects
Chromatography, biology, Chemistry, Euphorbia characias, Carbonic anhydrase activity, Cell Biology, Mass spectrometry, Photosynthesis, biology.organism_classification, Biochemistry, Mass spectrometric, Total inorganic carbon, Respiration, Hydration reaction, Molecular Biology
Abstract

The chemical forms of inorganic carbon, CO2 or HCO3-, incorporated during photosynthesis in photoautotrophic Euphorbia characias cell suspension cultures were determined in experiments using 13CO2 and a mass spectrometry technique. From the equations of the CO2 hydration reaction, a kinetic model was first developed, and the effect of photosynthesis on the external CO2 concentration was simulated. It was predicted from this model that CO2 and HCO3- uptakes could be differentiated by recording only the CO2 variation rate in the external medium, successively in absence then in presence of an exogenous carbonic anhydrase activity. The results obtained with either CO2-grown or air-grown photoautotrophic cells were in good agreement with the model and demonstrated that CO2 was the sole species taken up during photosynthesis. In addition no accumulation of inorganic carbon within the cells was observed in the light. Similarly, in dark, CO2 was the only species released by respiration in the external medium.

Published
1988

13. Cytoplasmic pH Regulation in Acer pseudoplatanus Cells

Author

Michel Pean, Yves Mathieu, Jean-Claude Beloeil, Jean-Yves Lallemand, Corinne Pasquier, and Jean Guern

Subjects
biology, Physiology, Plant Science, Nuclear magnetic resonance spectroscopy, Acer pseudoplatanus, Catharanthus roseus, Plant cell, biology.organism_classification, Acid load, chemistry.chemical_compound, chemistry, Biochemistry, Cytoplasm, Ph regulation, Genetics, Benzoic acid
Abstract

Modifications of cytoplasmic pH in Acer pseudoplatanus L. cells cultivated in suspension have been induced by acid-loads and studied by using 31P nuclear magnetic resonance spectroscopy. The initial drop of cytoplasmic pH, observed in the first minutes of exposure to weak lipophilic acids, was followed by a slow recovery to reach a plateau phase with a pH value lower than the initial one. Conversely, removal of the acid led to a sharp increase of cytoplasmic pH with in most cases an overshoot toward more alkaline values than the initial one and a subsequent decrease to more acidic values. This shows that A. pseudoplatanus cells powerfully regulate their cytoplasmic pH both on the acid side of their normal pH, during the acid-load, and on the alkaline side, after removal of acid. Similar results were obtained with different types of acid-loads, i.e. treatments with propionic or benzoic acid or bubbling with CO2-enriched air. This indicates that the occurrence of pH regulation does not depend upon the method used to acid-load the cells. The time courses of cytoplasmic pH observed for A. pseudoplatanus and also Catharanthus roseus cells are similar to those recorded for animal cells but different from those described for other plant materials for which no recovery phase was observed. This can be explained by different balances between the initial rate of proton influx brought in by the acids, and the capacity of proton consumption by the regulatory mechanisms. The existence of the recovery phase offers a unique possibility to study the regulation of the cytoplasmic pH of plant cells, as it has been done in animal systems.

Published
1986

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