44 results on '"Deloménie C"'
Search Results
2. Role of CinA in the regulation of inflammatory cytokines in keratinocytes: is Nrf2 the only regulator?
- Author
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Vallion, R., primary, Bouredji, A., additional, Damiens, M.-H., additional, Deloménie, C., additional, Marc, P., additional, Ferret, P.-J., additional, and Kerdine-Römer, S., additional
- Published
- 2021
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3. Action dichotomique du cinnamaldehyde dans l’activation des kératinocytes : un rôle à jouer pour Nrf2 ?
- Author
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Vallion, R., primary, Damiens, M.H., additional, Deloménie, C., additional, Pallardy, M., additional, Ferret, P.J., additional, and Kerdine-Römer, S., additional
- Published
- 2021
- Full Text
- View/download PDF
4. Beclin 1 and autophagy are required for the tumorigenicity of breast cancer stem-like/progenitor cells
- Author
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Gong, C, Bauvy, C, Tonelli, G, Yue, W, Deloménie, C, Nicolas, V, Zhu, Y, Domergue, V, Marin-Esteban, V, Tharinger, H, Delbos, L, Gary-Gouy, H, Morel, A-P, Ghavami, S, Song, E, Codogno, P, and Mehrpour, M
- Published
- 2013
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5. Increased Expression of MDR1 mRNAs and P-glycoprotein in Placentas from HIV-1 Infected Women
- Author
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Camus, M., Deloménie, C., Didier, N., Faye, A., Gil, S., Dauge, M.-C., Mabondzo, A., and Farinotti, R.
- Published
- 2006
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6. Fatty acid incorporation in endothelial cells and effects on endothelial nitric oxide synthase
- Author
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Couloubaly, S., Deloménie, C., Rousseau, D., Paul, J. L., Grynberg, A., and Pourci, M. L.
- Published
- 2007
7. 5-HT4 receptor agonists increase sAPPα levels in the cortex and hippocampus of male C57BL/6j mice
- Author
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Cachard-Chastel, M, Lezoualcʼh, F, Dewachter, I, Deloménie, C, Croes, S, Devijver, H, Langlois, M, Van Leuven, F, Sicsic, S, and Gardier, A M
- Published
- 2007
- Full Text
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8. Gene polymorphisms of MMPS-7, -9, -13 and -14 are predictors of aggravation in coronary artery disease?
- Author
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Ben Braiek, A., Baudin, B., Delomenie, C., Dumont, F., Chahed, H., Gamra, H., and Maatouk, F.
- Published
- 2018
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9. Homology modelling and structural analysis of human arylamine N-acetyltransferase NAT1: evidence for the conservation of a cysteine protease catalytic domain and an active-site loop
- Author
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Rodrigues-Lima, F, Deloménie, C, Goodfellow, G H, Grant, D M, and Dupret, J M
- Subjects
Models, Molecular ,Salmonella typhimurium ,Sequence Homology, Amino Acid ,Arylamine N-Acetyltransferase ,Protein Conformation ,Molecular Sequence Data ,Protein Structure, Secondary ,Protein Structure, Tertiary ,Isoenzymes ,Acetyltransferases ,Catalytic Domain ,Humans ,Amino Acid Sequence ,Conserved Sequence ,Research Article - Abstract
Arylamine N-acetyltransferases (EC 2.3.1.5) (NATs) catalyse the biotransformation of many primary arylamines, hydrazines and their N-hydroxylated metabolites, thereby playing an important role in both the detoxification and metabolic activation of numerous xenobiotics. The recently published crystal structure of the Salmonella typhimurium NAT (StNAT) revealed the existence of a cysteine protease-like (Cys-His-Asp) catalytic triad. In the present study, a three-dimensional homology model of human NAT1, based upon the crystal structure of StNAT [Sinclair, Sandy, Delgoda, Sim and Noble (2000) Nat. Struct. Biol. 7, 560-564], is demonstrated. Alignment of StNAT and NAT1, together with secondary structure predictions, have defined a consensus region (residues 29-131) in which 37% of the residues are conserved. Homology modelling provided a good quality model of the corresponding region in human NAT1. The location of the catalytic triad was found to be identical in StNAT and NAT1. Comparison of active-site structural elements revealed that a similar length loop is conserved in both species (residues 122-131 in NAT1 model and residues 122-133 in StNAT). This observation may explain the involvement of residues 125, 127 and 129 in human NAT substrate selectivity. Our model, and the fact that cysteine protease inhibitors do not affect the activity of NAT1, suggests that human NATs may have adapted a common catalytic mechanism from cysteine proteases to accommodate it for acetyl-transfer reactions.
- Published
- 2001
10. Beclin 1 and autophagy are required for the tumorigenicity of breast cancer stem-like/progenitor cells
- Author
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Gong, C, primary, Bauvy, C, additional, Tonelli, G, additional, Yue, W, additional, Deloménie, C, additional, Nicolas, V, additional, Zhu, Y, additional, Domergue, V, additional, Marin-Esteban, V, additional, Tharinger, H, additional, Delbos, L, additional, Gary-Gouy, H, additional, Morel, A-P, additional, Ghavami, S, additional, Song, E, additional, Codogno, P, additional, and Mehrpour, M, additional
- Published
- 2012
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11. Errata
- Author
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Garnier, A., primary, Fortin, D., additional, Deloménie, C., additional, Momken, I., additional, Veksler, V., additional, and Ventura‐Clapier, R., additional
- Published
- 2003
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12. Depressed mitochondrial transcription factors and oxidative capacity in rat failing cardiac and skeletal muscles
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Garnier, A., primary, Fortin, D., additional, Deloménie, C., additional, Momken, I., additional, Veksler, V., additional, and Ventura‐Clapier, R., additional
- Published
- 2003
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13. Les arylamine N-acétyltransférases : du polymorphisme génétique à la susceptibilité aux xénobiotiques.
- Author
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Deloménie, C, primary, Grant, DM, additional, Krishnamoorthy, R, additional, and Dupret, JM, additional
- Published
- 1998
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14. 5-HT4 receptor agonists increase sAPPα levels in the cortex and hippocampus of male C57BL/6j mice.
- Author
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Cachard-Chastel, M., Lezoualc'h, F., Dewachter, I., Deloménie, C., Croes, S., Devijver, H., Langlois, M., Van Leuven, F., Sicsic, S., and Gardier, A. M.
- Subjects
ALZHEIMER'S disease treatment ,AMYLOID beta-protein precursor ,SEROTONIN agonists ,ACETYLCHOLINESTERASE ,HIPPOCAMPUS (Brain) ,TRANSGENIC mice - Abstract
Background and purpose:A strategy to treat Alzheimer's disease (AD) is to increase the soluble form of amyloid precursor protein (sAPPα), a promnesic protein, in the brain. Because strong evidence supports beneficial effects of 5-hydroxytryptamine 5-HT
4 receptor agonists in memory and learning, we investigated the role of 5-HT4 receptors on APP processing in 8 weeks-old male C57BL/6j mice.Experimental approach:Mice were given, subcutaneously, prucalopride or ML 10302 (s.c.), two highly selective 5-HT4 receptor agonists and, up to 240 min later, the hippocampus and cortex were analysed by Western blot for sAPPα determination.Key results:Prucalopride (5 or 10 mg kg-1 ) significantly increased sAPPα levels in the hippocampus and cortex, but did not modify the expression level of APP mRNA as detected by quantitative RT-PCR. A selective 5-HT4 receptor antagonist, GR125487 (1 mg kg-1 , s.c.) inhibited prucalopride induced- increase in sAPPα levels. In addition, levels of sAPPα were increased by ML10302 only at 20 mg kg-1 and was limited to the cortex. Also, prucalopride increased sAPPα levels in the cortex of a transgenic mouse model of AD, expressing the London mutation of APP. Furthermore, the combined injection of a selective acetylcholinesterase inhibitor, donepezil and prucalopride induced a synergic increase in sAPPα levels in the cortex and hippocampus.Conclusions and implications:Our results demonstrate that the 5-HT4 receptor plays a key role in the non-amyloidogenic pathway of APP metabolism in vivo and give support to the beneficial use of 5-HT4 agonists for AD treatment.British Journal of Pharmacology (2007) 150, 883–892. doi:10.1038/sj.bjp.0707178; published online 26 February 2007 [ABSTRACT FROM AUTHOR]- Published
- 2007
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15. P15-24 Contact sensitizers modulate non-coding RNA expression and secretion of proinflammatory mediators in human keratinocytes.
- Author
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Paulet, V., Hardonnière, K., Deloménie, C., and Kerdine-Römer, S.
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- *
GENE expression , *ALLERGENS , *NON-coding RNA , *KERATINOCYTES , *SECRETION - Published
- 2024
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16. Genotyping of the polymorphic N-acetyltransferase (NAT2∗) gene locus in two native African populations.
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Deloménie, C, Sica, L, Grant, D M, Krishnamoorthy, R, and Dupret, J -M
- Published
- 1996
17. Intracellular mechanisms involved in cytokine modulation of endothelial cell functions
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Delomenie, C., Wautier, M.P., and Wautier, J.L.
- Published
- 1992
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18. Investigation of the mechanism of the antiproliferative effect of fendiline
- Author
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Deneulin, A., John, G.W., Shrivastava, R., Chevalier, A., Delomenie, C., and Massingham, R.
- Published
- 1991
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19. Multicellular tumor spheroid model to study the multifaceted role of tumor-associated macrophages in PDAC.
- Author
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Bidan N, Dunsmore G, Ugrinic M, Bied M, Moreira M, Deloménie C, Ginhoux F, Blériot C, de la Fuente M, and Mura S
- Subjects
- Humans, Tumor Microenvironment drug effects, Carcinoma, Pancreatic Ductal immunology, Carcinoma, Pancreatic Ductal pathology, Carcinoma, Pancreatic Ductal drug therapy, Cell Line, Tumor, Coculture Techniques, Cell Movement drug effects, Monocytes immunology, Vitamin E administration & dosage, Spheroids, Cellular drug effects, Pancreatic Neoplasms immunology, Pancreatic Neoplasms pathology, Pancreatic Neoplasms drug therapy, Tumor-Associated Macrophages immunology, Tumor-Associated Macrophages drug effects
- Abstract
While considerable efforts have been made to develop new therapies, progress in the treatment of pancreatic cancer has so far fallen short of patients' expectations. This is due in part to the lack of predictive in vitro models capable of accounting for the heterogeneity of this tumor and its low immunogenicity. To address this point, we have established and characterized a 3D spheroid model of pancreatic cancer composed of tumor cells, cancer-associated fibroblasts, and blood-derived monocytes. The fate of the latter has been followed from their recruitment into the tumor spheroid to their polarization into a tumor-associated macrophage (TAM)-like population, providing evidence for the formation of an immunosuppressive microenvironment.This 3D model well reproduced the multiple roles of TAMs and their influence on drug sensitivity and cell migration. Furthermore, we observed that lipid-based nanosystems consisting of sphingomyelin and vitamin E could affect the phenotype of macrophages, causing a reduction of characteristic markers of TAMs. Overall, this optimized triple coculture model gives a valuable tool that could find useful application for a more comprehensive understanding of TAM plasticity as well as for more predictive drug screening. This could increase the relevance of preclinical studies and help identify effective treatments., (© 2023. Controlled Release Society.)
- Published
- 2024
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20. A complex relation between levels of adult hippocampal neurogenesis and expression of the immature neuron marker doublecortin.
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Mendez-David I, David DJ, Deloménie C, Tritschler L, Beaulieu JM, Colle R, Corruble E, Gardier AM, and Hen R
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- Animals, Mice, Antidepressive Agents pharmacology, Hippocampus physiology, Neurogenesis physiology, Neurons, Doublecortin Protein, Fluoxetine pharmacology
- Abstract
We investigated the mechanisms underlying the effects of the antidepressant fluoxetine on behavior and adult hippocampal neurogenesis (AHN). After confirming our earlier report that the signaling molecule β-arrestin-2 (β-Arr2) is required for the antidepressant-like effects of fluoxetine, we found that the effects of fluoxetine on proliferation of neural progenitors and survival of adult-born granule cells are absent in the β-Arr2 knockout (KO) mice. To our surprise, fluoxetine induced a dramatic upregulation of the number of doublecortin (DCX)-expressing cells in the β-Arr2 KO mice, indicating that this marker can be increased even though AHN is not. We discovered two other conditions where a complex relationship occurs between the number of DCX-expressing cells compared to levels of AHN: a chronic antidepressant model where DCX is upregulated and an inflammation model where DCX is downregulated. We concluded that assessing the number of DCX-expressing cells alone to quantify levels of AHN can be complex and that caution should be applied when label retention techniques are unavailable., (© 2023 Wiley Periodicals LLC.)
- Published
- 2023
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21. Modulation of cardiac cAMP signaling by AMPK and its adjustments in pressure overload-induced myocardial dysfunction in rat and mouse.
- Author
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Garnier A, Leroy J, Deloménie C, Mateo P, Viollet B, Veksler V, Mericskay M, Ventura-Clapier R, and Piquereau J
- Subjects
- Mice, Rats, Animals, Calcium, Myocytes, Cardiac, Adrenergic Agents, Calcium, Dietary, AMP-Activated Protein Kinases, Heart Failure
- Abstract
The beta-adrenergic system is a potent stimulus for enhancing cardiac output that may become deleterious when energy metabolism is compromised as in heart failure. We thus examined whether the AMP-activated protein kinase (AMPK) that is activated in response to energy depletion may control the beta-adrenergic pathway. We studied the cardiac response to beta-adrenergic stimulation of AMPKα2-/- mice or to pharmacological AMPK activation on contractile function, calcium current, cAMP content and expression of adenylyl cyclase 5 (AC5), a rate limiting step of the beta-adrenergic pathway. In AMPKα2-/- mice the expression of AC5 (+50%), the dose response curve of left ventricular developed pressure to isoprenaline (p<0.001) or the response to forskolin, an activator of AC (+25%), were significantly increased compared to WT heart. Similarly, the response of L-type calcium current to 3-isobutyl-l-methylxanthine (IBMX), a phosphodiesterase inhibitor was significantly higher in KO (+98%, p<0.01) than WT (+57%) isolated cardiomyocytes. Conversely, pharmacological activation of AMPK by 5-aminoimidazole-4-carboxamide riboside (AICAR) induced a 45% decrease in AC5 expression (p<0.001) and a 40% decrease of cAMP content (P<0.001) as measured by fluorescence resonance energy transfer (FRET) compared to unstimulated rat cardiomyocytes. Finally, in experimental pressure overload-induced cardiac dysfunction, AMPK activation was associated with a decreased expression of AC5 that was blunted in AMPKα2-/- mice. The results show that AMPK activation down-regulates AC5 expression and blunts the beta-adrenergic cascade. This crosstalk between AMPK and beta-adrenergic pathways may participate in a compensatory energy sparing mechanism in dysfunctional myocardium., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Garnier et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2023
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22. The Inflammatory Response in Human Keratinocytes Exposed to Cinnamaldehyde Is Regulated by Nrf2.
- Author
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Vallion R, Hardonnière K, Bouredji A, Damiens MH, Deloménie C, Pallardy M, Ferret PJ, and Kerdine-Römer S
- Abstract
Keratinocytes (KC) play a crucial role in epidermal barrier function, notably through their metabolic activity and the detection of danger signals. Chemical sensitizers are known to activate the transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf2), leading to cellular detoxification and suppressed proinflammatory cytokines such as IL-1β, a key cytokine in skin allergy. We investigated the role of Nrf2 in the control of the proinflammatory response in human KC following treatment with Cinnamaldehyde (CinA), a well-known skin sensitizer. We used the well-described human KC cell line KERTr exposed to CinA. Our results showed that 250 μM of CinA did not induce any Nrf2 accumulation but increased the expression of proinflammatory cytokines. In contrast, 100 μM of CinA induced a rapid accumulation of Nrf2, inhibited IL-1β transcription, and downregulated the zymosan-induced proinflammatory response. Moreover, Nrf2 knockdown KERTr cells (KERTr ko) showed an increase in proinflammatory cytokines. Since the inhibition of Nrf2 has been shown to alter cellular metabolism, we performed metabolomic and seahorse analyses. The results showed a decrease in mitochondrial metabolism following KERTr ko exposure to CinA 100 µM. In conclusion, the fate of Nrf2 controls proinflammatory cytokine production in KCs that could be linked to its capacity to preserve mitochondrial metabolism upon chemical sensitizer exposure.
- Published
- 2022
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23. Topically Applied Chitosan-Coated Poly(isobutylcyanoacrylate) Nanoparticles Are Active Against Cutaneous Leishmaniasis by Accelerating Lesion Healing and Reducing the Parasitic Load.
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Malli S, Pomel S, Ayadi Y, Deloménie C, Da Costa A, Loiseau PM, and Bouchemal K
- Abstract
Parenteral administration of amphotericin B-deoxycholate (AmB-DOC) or pentavalent antimonials to cure cutaneous leishmaniasis (CL) results in severe adverse reactions, while topically applied antileishmanial drugs are ineffective despite their good tolerance. This work is aimed to investigate whether poly(isobutylcyanoacrylate) nanoparticles coated with chitosan (Cs-NPs) could provide intrinsic antileishmanial activity after topical application. In vitro evaluations revealed that nanoparticles were active against the promastigote, axenic amastigote, and intramacrophage forms of Leishmania major . In vivo evaluations after repetitive topical applications on the skin of mice infected with L. major showed that Cs-NPs combined or not with AmB-DOC allowed partial healing of the lesion characterized by histological analyses. The parasitic load of skin specimens collected from mice was significantly reduced compared with that from nontreated mice, as analyzed by quantitative polymerase chain reaction (q-PCR). Ultrastructure characterizations by electron microscopy of L. major promastigotes after incubation with Cs-NPs showed morphological alterations, including aberrant shape and swelling of mitochondria and parasitic vacuoles.
- Published
- 2019
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24. Aptamer-guided siRNA-loaded nanomedicines for systemic gene silencing in CD-44 expressing murine triple-negative breast cancer model.
- Author
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Alshaer W, Hillaireau H, Vergnaud J, Mura S, Deloménie C, Sauvage F, Ismail S, and Fattal E
- Subjects
- Animals, Cell Line, Tumor, Female, Gene Silencing, Humans, Liposomes, Luciferases genetics, Mice, Nanomedicine, Triple Negative Breast Neoplasms therapy, Aptamers, Nucleotide administration & dosage, Biomarkers, Tumor genetics, Hyaluronan Receptors genetics, RNA, Small Interfering administration & dosage, Triple Negative Breast Neoplasms genetics
- Abstract
In this study, we describe a liposome-based siRNA delivery system with a core composed of siRNA:protamine complex and a shell designed for the active targeting of CD44-expressing cells using for the first time the anti-CD44 aptamer (named Apt1) as targeting ligand. Among all functions, CD44 is the most common cancer stem cell surface biomarker and is found overexpressed in many tumors making this an attractive receptor for therapeutic targeting. This unique non-cationic system was evaluated for the silencing of the reporter gene of luciferase (luc2) in a triple-negative breast cancer model in vitro and in vivo. We show the possibility of conjugating an aptamer to siRNA-containing liposomes for an efficient gene silencing in CD44-expressing tumor cells in vivo, in the perspective of silencing disease-related genes in tumors., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
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25. Dendritic cells' death induced by contact sensitizers is controlled by Nrf2 and depends on glutathione levels.
- Author
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El Ali Z, Deloménie C, Botton J, Pallardy M, and Kerdine-Römer S
- Subjects
- Acrolein toxicity, Animals, Cell Death drug effects, Cell Death physiology, Dendritic Cells drug effects, Humans, Mice, Mice, Inbred C57BL, Mice, Knockout, NF-E2-Related Factor 2 deficiency, Acrolein analogs & derivatives, Dendritic Cells metabolism, Dinitrochlorobenzene toxicity, Glutathione metabolism, Haptens toxicity, NF-E2-Related Factor 2 physiology
- Abstract
Dendritic cells (DC) are known to play a major role during contact allergy induced by contact sensitizers (CS). Our previous studies showed that Nrf2 was induced in DC and controlled allergic skin inflammation in mice in response to chemicals. In this work, we raised the question of the role of Nrf2 in response to a stress provoked by chemical sensitizers in DC. We used two well-described chemical sensitizers, dinitrochlorobenzene (DNCB) and cinnamaldehyde (CinA), known to have different chemical reactivity and mechanism of action. First, we performed a RT-qPCR array showing that CinA was a higher inducer of immune and detoxification genes compared to DNCB. Interestingly, in the absence of Nrf2, gene expression was dramatically affected in response to DNCB but was slightly affected in response to CinA. These observations prompted us to study DC's cell death in response to both chemicals. DNCB and CinA increased apoptotic cells and decreased living cells in the absence of Nrf2. The characterization of DC apoptosis induced by both CS involved the mitochondrial-dependent caspase pathway and was regulated via Nrf2 in response to both chemicals. Oxidative stress induced by DNCB, and leading to cell death, was regulated by Nrf2. Unlike CinA, DNCB treatment provoked a significant reduction of intracellular GSH levels and up-regulated bcl-2 gene expression, under the control of Nrf2. This work underlies that chemical reactivity may control Nrf2-dependent gene expression leading to different cytoprotective mechanisms in DC., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
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26. Hyaluronic acid-conjugated lipoplexes for targeted delivery of siRNA in a murine metastatic lung cancer model.
- Author
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Leite Nascimento T, Hillaireau H, Vergnaud J, Rivano M, Deloménie C, Courilleau D, Arpicco S, Suk JS, Hanes J, and Fattal E
- Subjects
- Animals, Cell Line, Tumor, Female, Humans, Hyaluronan Receptors metabolism, Luciferases metabolism, Mice, RNA, Messenger metabolism, Hyaluronic Acid chemistry, Liposomes chemistry, Lung Neoplasms drug therapy, RNA, Small Interfering administration & dosage, RNA, Small Interfering chemistry
- Abstract
We have investigated the impact of hyaluronic acid (HA)-coating on the targeting capacity of siRNA lipoplexes to CD44-overexpressing tumor cells. Cellular uptake and localization of HA-lipoplexes were evaluated by flow cytometry and fluorescence microscopy and both methods showed that these lipoplexes were rapidly internalized and localized primarily within the cytoplasm. Inhibition of luciferase expression on the A549-luciferase lung cancer cell line was achieved in vitro using an anti-Luc siRNA. 81% of luciferase gene expression inhibition was obtained in vitro with HA-lipoplexes at +/- ratio 2. In vivo, in a murine A549 metastatic lung cancer model, the treatment with HA-lipoplexes carrying anti-luciferase siRNA led to a statistically significant decrease of luciferase expression as opposed to progressive increase with non-modified lipoplexes or NaCl 0.9%. The reduction of the expression of luciferase mRNA tumor of mice treated with HA-lipoplexes supported the inhibition effect due to siRNA. These results highlight the potential of HA-lipoplexes in CD44-targeting siRNA delivery., (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Published
- 2016
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27. Development and validation of a custom microarray for global transcriptome profiling of the fungus Aspergillus nidulans.
- Author
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Deloménie C, Grentzmann G, Oestreicher N, Mesnage R, and Vélot C
- Subjects
- Computational Biology methods, Gene Expression Profiling standards, Genomics methods, Genomics standards, Oligonucleotide Array Sequence Analysis methods, Oligonucleotide Array Sequence Analysis standards, Reproducibility of Results, Sensitivity and Specificity, Signal-To-Noise Ratio, Aspergillus nidulans genetics, Gene Expression Profiling methods, Transcriptome
- Abstract
Transcriptome profiling is a powerful tool for identifying gene networks from whole genome expression analysis in many living species. Here is described the first extensively characterized platform using Agilent microarray technology for transcriptome analysis in the filamentous fungus Aspergillus (Emericella) nidulans. We developed and validated a reliable gene expression microarray in 8 × 15 K format, with predictive and experimental data establishing its specificity and sensitivity. Either one or two 60-mer oligonucleotide probes were selected for each of 10,550 nuclear as well as 20 mitochondrial coding sequences. More than 99 % of probes were predicted to hybridize with 100 % identity to their aimed specific A. nidulans target only. Probe sensitivity was supported by a highly narrow distribution of melting temperatures together with thermodynamic features, which strongly favored probe-target perfect match hybridization, in comparison with predicted secondary structures. Array quality was evaluated through transcriptome comparison of two A. nidulans strains, differing by the presence or not of Escherichia coli LacZ transgene. High signal-to-noise ratios were measured, and signal reproducibility was established at intra-probe and inter-probe levels. Reproducibility of microarray performances was assessed by high correlation between two-color dye signals and between technical replicates. Results were confirmed by RT-qPCR analysis on five genes. Though it covers 100 % of the A. nidulans targeted coding sequences, this low density array allows limited experimental costs and simplified data analysis process, making it suitable for studying gene expression in this model organism through large numbers of experimental conditions, in basic, biomedical or industrial microbiology research fields.
- Published
- 2016
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28. Carabin protects against cardiac hypertrophy by blocking calcineurin, Ras, and Ca2+/calmodulin-dependent protein kinase II signaling.
- Author
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Bisserier M, Berthouze-Duquesnes M, Breckler M, Tortosa F, Fazal L, de Régibus A, Laurent AC, Varin A, Lucas A, Branchereau M, Marck P, Schickel JN, Deloménie C, Cazorla O, Soulas-Sprauel P, Crozatier B, Morel E, Heymes C, and Lezoualc'h F
- Subjects
- Animals, Calcium-Calmodulin-Dependent Protein Kinase Type 2 antagonists & inhibitors, Cells, Cultured, Female, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Myocytes, Cardiac metabolism, Rats, Signal Transduction physiology, Calcineurin metabolism, Calcium-Calmodulin-Dependent Protein Kinase Type 2 metabolism, Cardiomegaly metabolism, Cardiomegaly prevention & control, GTPase-Activating Proteins biosynthesis, Genes, ras physiology
- Abstract
Background: Cardiac hypertrophy is an early hallmark during the clinical course of heart failure and is regulated by various signaling pathways. However, the molecular mechanisms that negatively regulate these signal transduction pathways remain poorly understood., Methods and Results: Here, we characterized Carabin, a protein expressed in cardiomyocytes that was downregulated in cardiac hypertrophy and human heart failure. Four weeks after transverse aortic constriction, Carabin-deficient (Carabin(-/-)) mice developed exaggerated cardiac hypertrophy and displayed a strong decrease in fractional shortening (14.6±1.6% versus 27.6±1.4% in wild type plus transverse aortic constriction mice; P<0.0001). Conversely, compensation of Carabin loss through a cardiotropic adeno-associated viral vector encoding Carabin prevented transverse aortic constriction-induced cardiac hypertrophy with preserved fractional shortening (39.9±1.2% versus 25.9±2.6% in control plus transverse aortic constriction mice; P<0.0001). Carabin also conferred protection against adrenergic receptor-induced hypertrophy in isolated cardiomyocytes. Mechanistically, Carabin carries out a tripartite suppressive function. Indeed, Carabin, through its calcineurin-interacting site and Ras/Rab GTPase-activating protein domain, functions as an endogenous inhibitor of calcineurin and Ras/extracellular signal-regulated kinase prohypertrophic signaling. Moreover, Carabin reduced Ca(2+)/calmodulin-dependent protein kinase II activation and prevented nuclear export of histone deacetylase 4 after adrenergic stimulation or myocardial pressure overload. Finally, we showed that Carabin Ras-GTPase-activating protein domain and calcineurin-interacting domain were both involved in the antihypertrophic action of Carabin., Conclusions: Our study identifies Carabin as a negative regulator of key prohypertrophic signaling molecules, calcineurin, Ras, and Ca(2+)/calmodulin-dependent protein kinase II and implicates Carabin in the development of cardiac hypertrophy and failure., (© 2014 American Heart Association, Inc.)
- Published
- 2015
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29. Disturbed intestinal nitrogen homeostasis in a mouse model of high-fat diet-induced obesity and glucose intolerance.
- Author
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Do TT, Hindlet P, Waligora-Dupriet AJ, Kapel N, Neveux N, Mignon V, Deloménie C, Farinotti R, Fève B, and Buyse M
- Subjects
- Allostasis, Amino Acid Transport Systems genetics, Amino Acid Transport Systems metabolism, Amino Acids blood, Animals, DNA analysis, Diet, High-Fat adverse effects, Dipeptidyl Peptidase 4 chemistry, Dipeptidyl Peptidase 4 genetics, Dipeptidyl Peptidase 4 metabolism, Feces chemistry, Feces microbiology, Gene Expression Regulation, Glucose Intolerance etiology, Glucose Intolerance microbiology, Glucose Intolerance pathology, Gram-Negative Bacteria growth & development, Gram-Negative Bacteria isolation & purification, Gram-Positive Bacteria growth & development, Gram-Positive Bacteria isolation & purification, Intestinal Mucosa microbiology, Intestinal Mucosa pathology, Intestines microbiology, Intestines pathology, Male, Mice, Mice, Inbred C57BL, Nitrogen analysis, Nitrogen metabolism, Obesity etiology, Obesity microbiology, Obesity pathology, Peptide Transporter 1, Symporters genetics, Symporters metabolism, Amino Acid Transport Systems biosynthesis, Amino Acids metabolism, Disease Models, Animal, Glucose Intolerance metabolism, Intestinal Absorption, Intestinal Mucosa metabolism, Obesity metabolism, Symporters biosynthesis
- Abstract
The oligopeptide transporter peptide cotransporter-1 Slc15a1 (PEPT1) plays a major role in the regulation of nitrogen supply, since it is responsible for 70% of the dietary nitrogen absorption. Previous studies demonstrated that PEPT1 expression and function in jejunum are reduced in diabetes and obesity, suggesting a nitrogen malabsorption from the diet. Surprisingly, we reported here a decrease in gut nitrogen excretion in high-fat diet (HFD)-fed mice and further investigated the mechanisms that could explain this apparent contradiction. Upon HFD, mice exhibited an increased concentration of free amino acids (AAs) in the portal vein (60%) along with a selective increase in the expression of two AA transporters (Slc6a20a, Slc36a1), pointing to a specific and adaptive absorption of some AAs. A delayed transit time (+40%) and an increased intestinal permeability (+80%) also contribute to the increase in nitrogen absorption. Besides, HFD mice exhibited a 2.2-fold decrease in fecal DNA resulting from a reduction in nitrogen catabolism from cell desquamation and/or in the intestinal microbiota. Indeed, major quantitative (2.5-fold reduction) and qualitative alterations of intestinal microbiota were observed in feces of HFD mice. Collectively, our results strongly suggest that both increased AA transporters, intestinal permeability and transit time, and changes in gut microbiota are involved in the increased circulating AA levels. Modifications in nitrogen homeostasis provide a new insight in HFD-induced obesity and glucose intolerance; however, whether these modifications are beneficial or detrimental for the HFD-associated metabolic complications remains an open issue.
- Published
- 2014
- Full Text
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30. Ventral hippocampal molecular pathways and impaired neurogenesis associated with 5-HT₁A and 5-HT₁B receptors disruption in mice.
- Author
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Xia L, Deloménie C, David I, Rainer Q, Marouard M, Delacroix H, David DJ, Gardier AM, and Guilloux JP
- Subjects
- Animals, Cell Proliferation, Cell Survival, Gene Expression Profiling, Hippocampus cytology, Immunohistochemistry, Male, Mice, Mice, Knockout, Neurons cytology, Oligonucleotide Array Sequence Analysis, Real-Time Polymerase Chain Reaction, Receptor, Serotonin, 5-HT1A genetics, Receptor, Serotonin, 5-HT1B genetics, Hippocampus metabolism, Neurogenesis, Receptor, Serotonin, 5-HT1A physiology, Receptor, Serotonin, 5-HT1B physiology
- Abstract
The serotonergic system has been widely implicated in stress related psychiatric disorders such as depression and anxiety. Generation of receptor knockout mice has offered a new approach to study processes underlying anxiety. For instance, knockout mice for both 5-HT(1A) and 5-HT(1B) receptors (5-HT(1A/1B)(-/-)) display an anxious phenotype, associated with robust physiological and neurochemical changes related to brain serotonin function. As ventral hippocampus is a key region in the mediation and genesis of anxiety, we explored the transcriptome changes induced by the genetic inactivation of these two receptors in 5-HT(1A/1B)(-/-) mice. Dissociation of ventral vs. dorsal hippocampus was confirmed by the over-expression of selective markers in both regions. 723 genes were observed up/down regulated in 5-HT(1A/1B)(-/-) mice. Using Ingenuity, biological networks and signal transduction pathway analysis corresponding to the identified gene revealed putative dysregulation of nervous system development and function, especially genes associated with long-term potentiation and adult neurogenesis (including Bdnf, Camk2a, Camk4, and Klf9). Furthermore, immunohistochemistry experiments studying adult hippocampal neurogenesis in adult 5-HT(1A/1B)(-/-) mice showed a decreased survival, but not proliferation of newborn cells in our model., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2012
- Full Text
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31. Biodegradable nanoparticles meet the bronchial airway barrier: how surface properties affect their interaction with mucus and epithelial cells.
- Author
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Mura S, Hillaireau H, Nicolas J, Kerdine-Römer S, Le Droumaguet B, Deloménie C, Nicolas V, Pallardy M, Tsapis N, and Fattal E
- Subjects
- Cell Line, Electric Impedance, Epithelial Cells physiology, Gene Expression drug effects, Humans, Lactic Acid metabolism, Mucin 5AC genetics, Mucin 5AC metabolism, Permeability, Polyglycolic Acid metabolism, Polylactic Acid-Polyglycolic Acid Copolymer, Surface Properties, Bronchi cytology, Epithelial Cells metabolism, Lactic Acid pharmacology, Mucus metabolism, Nanoparticles, Polyglycolic Acid pharmacology
- Abstract
Despite the wide interest raised by lung administration of nanoparticles (NPs) for the treatment of various diseases, little information is available on their effect toward the airway epithelial barrier function. In this study, the potential damage of the pulmonary epithelium upon exposure to poly(lactide-co-glycolide) (PLGA) NPs has been assessed in vitro using a Calu-3-based model of the bronchial epithelial barrier. Positively and negatively charged as well as neutral PLGA NPs were obtained by coating their surface with chitosan (CS), poloxamer (PF68), or poly(vinyl alcohol) (PVA). The role of NP surface chemistry and charge on the epithelial resistance and mucus turnover, using MUC5AC as a marker, was investigated. The interaction with mucin reduced the penetration of CS- and PVA-coated NPs, while the hydrophilic PF68-coated NPs diffused across the mucus barrier leading to a higher intracellular accumulation. Only CS-coated NPs caused a transient but reversible decrease of the trans-epithelial electrical resistance (TEER). None of the NP formulations increased MUC5AC mRNA expression or the protein levels. These in vitro results highlight the safety of PLGA NPs toward the integrity and function of the bronchial airway barrier and demonstrate the crucial role of NP surface properties to achieve a controlled and sustained delivery of drugs via the pulmonary route.
- Published
- 2011
- Full Text
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32. Reduced intestinal absorption of dipeptides via PepT1 in mice with diet-induced obesity is associated with leptin receptor down-regulation.
- Author
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Hindlet P, Bado A, Kamenicky P, Deloménie C, Bourasset F, Nazaret C, Farinotti R, and Buyse M
- Subjects
- Animals, Biological Transport, Caco-2 Cells, Disease Models, Animal, Down-Regulation, Gastrointestinal Tract metabolism, Humans, Leptin metabolism, MAP Kinase Signaling System, Male, Mice, Obesity etiology, Peptide Transporter 1, RNA, Messenger biosynthesis, Time Factors, Diet adverse effects, Dipeptides metabolism, Intestinal Absorption, Obesity metabolism, Receptors, Leptin biosynthesis, Symporters biosynthesis
- Abstract
Leptin is a major determinant of energy homeostasis, acting both centrally and in the gastrointestinal tract. We previously reported that acute leptin treatment enhances the absorption of di- and tripeptides via the proton-dependent PepT1 transporter. In this study, we investigated the long term effect of leptin on PepT1 levels and activity in Caco2 cell monolayers in vitro. We then assessed the significance of the regulation of PepT1 in vivo in a model of diet-induced obesity. We demonstrated that 1) leptin regulated PepT1 at the transcriptional level, via the MAPK pathway, and at the translational level, via ribosomal protein S6 activation, in Caco2 cells and 2) this activation was systematically followed by a time- and concentration-dependent loss of leptin action reflecting desensitization. Deciphering this desensitization, we demonstrated that leptin induced a down-regulation of its own receptor protein and mRNA expression. More importantly, we showed, in mice with diet-induced obesity, that a 4-week hypercaloric diet resulted in a 46% decrease in PepT1-specific transport, because of a 30% decrease in PepT1 protein and a 50% decrease in PepT1 mRNA levels. As shown in Caco2 cells, these changes in PepT1 were supported by a parallel 2-fold decrease in leptin receptor expression in mice. Taken together, these results indicate that during induction of obesity, leptin resistance may also occur peripherally in the gastrointestinal tract, disrupting the absorption of oligopeptides and peptidomimetic drugs.
- Published
- 2009
- Full Text
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33. Consequences of changes in BDNF levels on serotonin neurotransmission, 5-HT transporter expression and function: studies in adult mice hippocampus.
- Author
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Deltheil T, Guiard BP, Guilloux JP, Nicolas L, Deloménie C, Repérant C, Le Maitre E, Leroux-Nicollet I, Benmansour S, Coudoré F, David DJ, and Gardier AM
- Subjects
- Animals, Antidepressive Agents pharmacology, Brain-Derived Neurotrophic Factor pharmacology, Citalopram metabolism, Mice, Paroxetine pharmacology, RNA, Messenger analysis, Serotonin Plasma Membrane Transport Proteins physiology, Brain-Derived Neurotrophic Factor physiology, Hippocampus metabolism, Microdialysis methods, Serotonin metabolism, Serotonin Plasma Membrane Transport Proteins genetics
- Abstract
In vivo intracerebral microdialysis is an important neurochemical technique that has been applied extensively in genetic and pharmacological studies aimed at investigating the relationship between neurotransmitters. Among the main interests of microdialysis application is the infusion of drugs through the microdialysis probe (reverse dialysis) in awake, freely moving animals. As an example of the relevance of intracerebral microdialysis, this review will focus on our recent neurochemical results showing the impact of Brain-Derived Neurotrophic Factor (BDNF) on serotonergic neurotransmission in basal and stimulated conditions. Indeed, although the elevation of 5-HT outflow induced by chronic administration of selective serotonin reuptake inhibitors (SSRIs) causes an increase in BDNF protein levels and expression (mRNA) in the hippocampus of rodents, the reciprocal interaction has not been demonstrated yet. Thus, the neurochemical sight of this question will be addressed here by examining the consequences of either a constitutive decrease or increase in brain BDNF protein levels on hippocampal extracellular levels of 5-HT in conscious mice.
- Published
- 2008
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34. Antibiotics involved in Clostridium difficile-associated disease increase colonization factor gene expression.
- Author
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Denève C, Deloménie C, Barc MC, Collignon A, and Janoir C
- Subjects
- Adhesins, Bacterial drug effects, Adhesins, Bacterial metabolism, Anti-Bacterial Agents pharmacology, Bacterial Adhesion drug effects, Bacterial Adhesion physiology, Bacterial Proteins drug effects, Bacterial Proteins genetics, Cell Line, Tumor, Clostridioides difficile drug effects, Clostridioides difficile genetics, Diarrhea microbiology, Drug Resistance, Gene Expression Regulation, Bacterial drug effects, Humans, Iron metabolism, Microbial Sensitivity Tests, Osmolar Concentration, Anti-Bacterial Agents adverse effects, Bacterial Proteins metabolism, Clostridioides difficile metabolism, Clostridium Infections microbiology, Diarrhea chemically induced, Gene Expression Regulation, Bacterial physiology
- Abstract
Clostridium difficile is the most common cause of antibiotic-associated diarrhoea. Antibiotics are presumed to disturb the normal intestinal microbiota, leading to depletion of the barrier effect and colonization by pathogenic bacteria. This first step of infection includes adherence to epithelial cells. We investigated the impact of various environmental conditions in vitro on the expression of genes encoding known, or putative, colonization factors: three adhesins, P47 (one of the two S-layer proteins), Cwp66 and Fbp68, and a protease, Cwp84. The conditions studied included hyperosmolarity, iron depletion and exposure to several antibiotics (ampicillin, clindamycin, ofloxacin, moxifloxacin and kanamycin). The analysis was performed on three toxigenic and three non-toxigenic C. difficile isolates using real-time PCR. To complete this work, the impact of ampicillin and clindamycin on the adherence of C. difficile to Caco-2/TC7 cells was analysed. Overall, for the six strains of C. difficile studied, exposure to subinhibitory concentrations (1/2 MIC) of clindamycin and ampicillin led to the increased expression of genes encoding colonization factors. This was correlated with the increased adherence of C. difficile to cultured cells under the same conditions. The levels of gene regulation observed among the six strains studied were highly variable, cwp84 being the most upregulated. In contrast, the expression of these genes was weakly, or not significantly, modified in the presence of ofloxacin, moxifloxacin or kanamycin. These results suggest that, in addition to the disruption of the normal intestinal microbiota and its barrier effect, the high propensity of antibiotics such as ampicillin and clindamycin to induce C. difficile infection could also be explained by their direct role in enhancing colonization by C. difficile.
- Published
- 2008
- Full Text
- View/download PDF
35. A new homolog of FocA transporters identified in cadmium-resistant Euglena gracilis.
- Author
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Deloménie C, Foti E, Floch E, Diderot V, Porquet D, Dupuy C, and Bonaly J
- Subjects
- Amino Acid Sequence, Animals, Molecular Sequence Data, Protein Isoforms chemistry, Protein Isoforms metabolism, Cadmium administration & dosage, Drug Resistance physiology, Escherichia coli Proteins chemistry, Escherichia coli Proteins metabolism, Euglena gracilis drug effects, Euglena gracilis metabolism, Membrane Transport Proteins chemistry, Membrane Transport Proteins metabolism
- Abstract
To better understand the cellular mechanism of stress resistance to various pollutants (cadmium, pentachlorophenol), we undertook a survey of the Euglena gracilis transcriptome by mRNA differential display and cDNA cloning. We performed a real-time RT-PCR analysis upon four selected genes. One of them significantly changed its expression level in response to stress treatments: B25 gene was overexpressed in Cd-resistant cells whereas it was down-regulated in PCP-adapted cells. By Race assays we obtained for B25 a 1093bp cDNA. The deduced protein was identified as a bacterial formate/nitrite transporter (FocA) homolog and the gene was named EgFth. From all the data, we concluded that EgFth overexpression was related to chronic exposure to cadmium.
- Published
- 2007
- Full Text
- View/download PDF
36. Peroxisome proliferator-activated receptor alpha physically interacts with CCAAT/enhancer binding protein (C/EBPbeta) to inhibit C/EBPbeta-responsive alpha1-acid glycoprotein gene expression.
- Author
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Mouthiers A, Baillet A, Deloménie C, Porquet D, and Mejdoubi-Charef N
- Subjects
- Animals, Dexamethasone pharmacology, Down-Regulation, Gene Expression Regulation drug effects, Glucocorticoids pharmacology, Nuclear Receptor Coactivator 2, Orosomucoid biosynthesis, Peroxisome Proliferators pharmacology, Promoter Regions, Genetic, Pyrimidines pharmacology, Rats, Rats, Sprague-Dawley, Transcription Factors metabolism, Transcription, Genetic drug effects, CCAAT-Enhancer-Binding Protein-beta metabolism, Gene Expression Regulation physiology, Orosomucoid genetics, PPAR alpha metabolism
- Abstract
Recently, the role of the peroxisome proliferator-activated receptor alpha (PPARalpha) in the hepatic inflammatory response has been associated to the decrease of acute phase protein transcription, although the molecular mechanisms are still to be elucidated. Here, we were interested in the regulation by Wy-14643 (PPARalpha agonist) of alpha1-acid glycoprotein (AGP), a positive acute phase protein, after stimulation by Dexamethasone (Dex), a major modulator of the inflammatory response. In cultured rat hepatocytes, we demonstrate that PPARalpha inhibits at the transcriptional level the Dex-induced AGP gene expression. PPARalpha exerts this inhibitory effect by antagonizing the CCAAT/enhancer binding protein (C/EBPbeta) transcription factor that is involved in Dex-dependent up-regulation of AGP gene expression. Overexpression of C/EBPbeta alleviates the repressive effect of PPARalpha, thus restoring the Dex-stimulated AGP promoter activity. Furthermore, glutathione-S-transferase GST pull-down and coimmunoprecipitation experiments evidenced, for the first time, a physical interaction between PPARalpha and the C-terminal DNA binding region of C/EBPbeta, thus preventing it from binding to specific sequence elements of the AGP promoter. Altogether, these results provide an additional molecular mechanism of negative regulation of acute phase protein gene expression by sequestration of the C/EBPbeta transcription factor by PPARalpha and reveal the high potency of the latter in controlling inflammation.
- Published
- 2005
- Full Text
- View/download PDF
37. Phosphatidylinositol 3-kinase and calcium-activated transcription pathways are required for VLDL-induced smooth muscle cell proliferation.
- Author
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Lipskaia L, Pourci ML, Deloménie C, Combettes L, Goudounèche D, Paul JL, Capiod T, and Lompré AM
- Subjects
- Active Transport, Cell Nucleus drug effects, Animals, Calcineurin metabolism, Calcium Signaling drug effects, Calcium Signaling physiology, Cell Division drug effects, Cell Division physiology, Cells, Cultured, Cyclic AMP Response Element-Binding Protein metabolism, DNA-Binding Proteins metabolism, Dose-Response Relationship, Drug, Enzyme Inhibitors pharmacology, Humans, Intracellular Fluid metabolism, Male, Muscle, Smooth, Vascular cytology, Muscle, Smooth, Vascular physiology, NFATC Transcription Factors, Phosphoinositide-3 Kinase Inhibitors, Phosphorylation drug effects, Rats, Rats, Wistar, Signal Transduction drug effects, Signal Transduction physiology, Transcription Factors metabolism, Calcium metabolism, Lipoproteins, VLDL pharmacology, Muscle, Smooth, Vascular drug effects, Nuclear Proteins, Phosphatidylinositol 3-Kinases metabolism, Transcription, Genetic physiology
- Abstract
Little is known regarding the molecular mechanisms of atherogenicity of triglyceride-rich lipoproteins such as very low-density lipoproteins (VLDLs). We examined the effect of VLDL on proliferation of rat aortic smooth muscle cells, intracellular Ca2+ handling, and activity of cAMP-responsive element binding protein (CREB) and nuclear factor of activated T cells (NFAT) transcription factors. VLDL, isolated from human serum, dose- and time-dependently promoted proliferation. After 4 hours of exposure to VLDL (0.15 g/L proteins), the caffeine-induced Ca2+ release was inhibited and the IP3-sensitive Ca2+ release induced by ATP (10 micromol/L) was markedly prolonged. In quiescent cells, CREB was phosphorylated (pCREB) and NFAT was present in the cytosol, whereas in cells exposed to VLDL for 4 to 24 hours, pCREB disappeared and NFAT was translocated to the nucleus. VLDL-induced NFAT translocation and proliferation were blocked by cyclosporin A and LY294002 involving calcineurin and phosphatidylinositol 3-kinase (PI3K) pathways. Indeed, VLDLs rapidly phosphorylate protein kinase B and glycogen synthase kinase-3beta in a PI3K-dependent way. These results provide the first evidence that VLDLs induce smooth muscle cell proliferation by activating the PI3K pathway and nuclear NFAT translocation. Blockade of the Ca2+-induced Ca2+ release mechanism and dephosphorylation of pCREB contribute but were not sufficient to induce a proliferating phenotype.
- Published
- 2003
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38. Association of GSTT1 non-null and NAT1 slow/rapid genotypes with von Hippel-Lindau tumour suppressor gene transversions in sporadic renal cell carcinoma.
- Author
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Gallou C, Longuemaux S, Deloménie C, Méjean A, Martin N, Martinet S, Palais G, Bouvier R, Droz D, Krishnamoorthy R, Junien C, Béroud C, and Dupret JM
- Subjects
- Adult, Aged, Chromosome Aberrations, Female, Frameshift Mutation, Gene Frequency, Genotype, Humans, Isoenzymes, Loss of Heterozygosity, Male, Middle Aged, Polymerase Chain Reaction, Polymorphism, Restriction Fragment Length, Polymorphism, Single-Stranded Conformational, Von Hippel-Lindau Tumor Suppressor Protein, Xenobiotics metabolism, Acetyltransferases genetics, Arylamine N-Acetyltransferase, Carcinoma, Renal Cell genetics, Genes, Tumor Suppressor, Glutathione Transferase genetics, Ligases, Mutation, Proteins genetics, Tumor Suppressor Proteins, Ubiquitin-Protein Ligases
- Abstract
The von Hippel-Lindau (VHL) tumour suppressor gene is commonly mutated in renal cell carcinoma of clear cell type (CCRCC). We investigated the possible relationship between VHL mutations in sporadic CCRCC and polymorphism of genes encoding enzymes involved in carcinogen metabolism: two cytochrome P450 monooxygenases (CYP1A1 and CYP2D6), one NAD[P]H:quinone oxidoreductase (NQO1), three glutathione S-transferases (GSTM1, GSTT1 and GSTP1) and two arylamine N-acetyltransferases (NAT1 and NAT2). We analysed DNA from tumour and nontumoural kidney tissue from 195 CCRCC patients. Single VHL mutations were identified in 88 patients and double mutations were present in two patients. Nine of 18 transversions were GC to TA, four were AT to TA, four were GC to CG and one was AT to CG. Ten of 19 transitions were GC to AT and nine were AT to GC. We also identified 53 frameshifts and two GC to AT at CpG. An excess of transversions was observed in a subset of patients with active GSTT1 [GSTT1 (+) genotype] and probably defective NAT1 (NAT1 S/R variant genotype). All 18 transversions were in GSTT1 (+) patients, whereas only 76% of transitions (P = 0.05) and 81% of the other mutations (P = 0.06) occurred in this genotype. We found that 28% of the transversions were in the NAT1 S/R genotype versus 12% of the transitions (P = 0.40) and 4% of the other mutations (P = 0.01). This suggests that pharmacogenetic polymorphisms may be associated with the type of acquired VHL mutation, which may modulate CCRCC development.
- Published
- 2001
- Full Text
- View/download PDF
39. Homology modelling and structural analysis of human arylamine N-acetyltransferase NAT1: evidence for the conservation of a cysteine protease catalytic domain and an active-site loop.
- Author
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Rodrigues-Lima F, Deloménie C, Goodfellow GH, Grant DM, and Dupret JM
- Subjects
- Acetyltransferases genetics, Acetyltransferases metabolism, Amino Acid Sequence, Arylamine N-Acetyltransferase chemistry, Arylamine N-Acetyltransferase genetics, Catalytic Domain, Conserved Sequence, Humans, Isoenzymes, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Structure, Secondary, Protein Structure, Tertiary, Salmonella typhimurium enzymology, Salmonella typhimurium genetics, Sequence Homology, Amino Acid, Acetyltransferases chemistry
- Abstract
Arylamine N-acetyltransferases (EC 2.3.1.5) (NATs) catalyse the biotransformation of many primary arylamines, hydrazines and their N-hydroxylated metabolites, thereby playing an important role in both the detoxification and metabolic activation of numerous xenobiotics. The recently published crystal structure of the Salmonella typhimurium NAT (StNAT) revealed the existence of a cysteine protease-like (Cys-His-Asp) catalytic triad. In the present study, a three-dimensional homology model of human NAT1, based upon the crystal structure of StNAT [Sinclair, Sandy, Delgoda, Sim and Noble (2000) Nat. Struct. Biol. 7, 560-564], is demonstrated. Alignment of StNAT and NAT1, together with secondary structure predictions, have defined a consensus region (residues 29-131) in which 37% of the residues are conserved. Homology modelling provided a good quality model of the corresponding region in human NAT1. The location of the catalytic triad was found to be identical in StNAT and NAT1. Comparison of active-site structural elements revealed that a similar length loop is conserved in both species (residues 122-131 in NAT1 model and residues 122-133 in StNAT). This observation may explain the involvement of residues 125, 127 and 129 in human NAT substrate selectivity. Our model, and the fact that cysteine protease inhibitors do not affect the activity of NAT1, suggests that human NATs may have adapted a common catalytic mechanism from cysteine proteases to accommodate it for acetyl-transfer reactions.
- Published
- 2001
- Full Text
- View/download PDF
40. Identification and functional characterization of arylamine N-acetyltransferases in eubacteria: evidence for highly selective acetylation of 5-aminosalicylic acid.
- Author
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Deloménie C, Fouix S, Longuemaux S, Brahimi N, Bizet C, Picard B, Denamur E, and Dupret JM
- Subjects
- Acetylation, Arylamine N-Acetyltransferase antagonists & inhibitors, Arylamine N-Acetyltransferase classification, Arylamine N-Acetyltransferase genetics, Blotting, Southern, DNA, Bacterial analysis, Humans, Kinetics, Polymerase Chain Reaction, Proteobacteria growth & development, Arylamine N-Acetyltransferase metabolism, Colon microbiology, Mesalamine metabolism, Proteobacteria enzymology
- Abstract
Arylamine N-acetyltransferase activity has been described in various bacterial species. Bacterial N-acetyltransferases, including those from bacteria of the gut flora, may be involved in the metabolism of xenobiotics, thereby exerting physiopathological effects. We characterized these enzymes further by steady-state kinetics, time-dependent inhibition, and DNA hybridization in 40 species, mostly from the human intestinal microflora. We report for the first time N-acetyltransferase activity in 11 species of Proteobacteriaceae from seven genera: Citrobacter amalonaticus, Citrobacter farmeri, Citrobacter freundii, Klebsiella ozaenae, Klebsiella oxytoca, Klebsiella rhinoscleromatis, Morganella morganii, Serratia marcescens, Shigella flexneri, Plesiomonas shigelloides, and Vibrio cholerae. We estimated apparent kinetic parameters and found that 5-aminosalicylic acid, a compound efficient in the treatment of inflammatory bowel diseases, was acetylated with a catalytic efficiency 27 to 645 times higher than that for its isomer, 4-aminosalicylic acid. In contrast, para-aminobenzoic acid, a folate precursor in bacteria, was poorly acetylated. Of the wild-type strains studied, Pseudomonas aeruginosa was the best acetylator in terms of both substrate spectrum and catalytic efficiency. DNA hybridization with a Salmonella enterica serovar Typhimurium-derived probe suggested the presence of this enzyme in eight proteobacterial and four gram-positive species. Molecular aspects together with the kinetic data suggest distinct functional features for this class of microbial enzymes.
- Published
- 2001
- Full Text
- View/download PDF
41. Candidate genetic modifiers of individual susceptibility to renal cell carcinoma: a study of polymorphic human xenobiotic-metabolizing enzymes.
- Author
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Longuemaux S, Deloménie C, Gallou C, Méjean A, Vincent-Viry M, Bouvier R, Droz D, Krishnamoorthy R, Galteau MM, Junien C, Béroud C, and Dupret JM
- Subjects
- Adult, Alleles, Arylamine N-Acetyltransferase genetics, Carcinoma, Renal Cell enzymology, Case-Control Studies, Cytochrome P-450 CYP1A1 genetics, Cytochrome P-450 CYP2D6 genetics, Female, Gene Frequency, Genotype, Glutathione Transferase genetics, Humans, Inactivation, Metabolic, Kidney Neoplasms enzymology, Male, Middle Aged, Polymerase Chain Reaction, Risk Factors, Carcinoma, Renal Cell genetics, Cytochrome P-450 Enzyme System genetics, Genetic Predisposition to Disease, Kidney Neoplasms genetics, Polymorphism, Restriction Fragment Length, Xenobiotics metabolism
- Abstract
The steady increase in sporadic renal cell carcinoma (RCC) observed in industrialized countries supports the notion that certain carcinogens present in the environment (tobacco smoke, drugs, pollutants, and dietary constituents) may affect the occurrence of RCC. Many of the enzymes dealing with such environmental factors are polymorphic and may, therefore, confer variable susceptibility to RCC. This case-control study was designed to test for an association between genetic polymorphism of enzymes involved in xenobiotic metabolism and the risk of sporadic RCC. Genomic DNA was obtained from 173 patients with RCC and 211 controls of Caucasian origin. We used PCR-RFLP to investigate polymorphism for the most common alleles at two cytochrome-P450 mono-oxygenases (CYP1A1 and CYP2D6), one NAD[P]H:quinone oxidoreductase (NQO1), three glutathione S-transferases (GSTM1, GSTT1, and GSTP1), and one N-acetyltransferase (NAT2) loci. The CYP1A1 (m) "variant" genotype, which contains at least one copy of the CYP1A1 variant alleles, was found to be associated with a 2.1-fold [95% confidence interval (CI), 1.1-3.9] increase in the risk of RCC. There was also a higher risk of RCC for subjects with the CYP1A1 (m) variant genotype combined with any of the following genotypes: GSTT1 (+) "active" [odds ratio (OR), 2.3; 95% CI, 1.2-4.5], GSTP1 (m) variant (OR, 2.4; 95% CI, 1.0-5.4), or NAT2 (-) "slow acetylator" (OR, 2.5; 95% CI, 1.1-5.5). A significant association was also found for the GSTM1 (-) "null" and GSTP1 (m) genotypes combined with either NAT2 (-) (OR, 2.6; 95% CI, 1.2-5.8) or CYP1A1 (m) (OR, 3.5; 95% CI, 1.1-11.2). The CYP2D6 (-) "poor metabolizer " and the NQO1 (-) "defective" genotypes were not clearly associated with a higher risk of RCC. Our data demonstrate for the first time a significant association between a group of pharmacogenetic polymorphisms and RCC risk. These positive findings suggest that interindividual variation in the metabolic pathways involved in the functionalization and detoxification of specific xenobiotics is an important susceptibility factor for RCC in Caucasians.
- Published
- 1999
42. Glutathione S-transferase (GSTM1) null genotype and sulphonamide intolerance in acquired immunodeficiency syndrome.
- Author
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Deloménie C, Mathelier-Fusade P, Longuemaux S, Rozenbaum W, Leynadier F, Krishnamoorthy R, and Dupret JM
- Subjects
- Adult, Genotype, Humans, Middle Aged, Risk Factors, Acquired Immunodeficiency Syndrome enzymology, Acquired Immunodeficiency Syndrome genetics, Drug Hypersensitivity enzymology, Drug Hypersensitivity genetics, Glutathione Transferase deficiency, Glutathione Transferase genetics, Sulfonamides adverse effects
- Published
- 1997
- Full Text
- View/download PDF
43. Study of the role of the highly conserved residues Arg9 and Arg64 in the catalytic function of human N-acetyltransferases NAT1 and NAT2 by site-directed mutagenesis.
- Author
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Deloménie C, Goodfellow GH, Krishnamoorthy R, Grant DM, and Dupret JM
- Subjects
- Amino Acid Sequence, Arylamine N-Acetyltransferase genetics, Arylamine N-Acetyltransferase metabolism, Conserved Sequence, Cystine, Enzyme Stability, Humans, Hydrogen-Ion Concentration, Isoenzymes genetics, Isoenzymes metabolism, Molecular Sequence Data, Mutagenesis, Site-Directed, Phenylglyoxal pharmacology, Protein Denaturation, Sequence Alignment, Structure-Activity Relationship, Arginine, Arylamine N-Acetyltransferase chemistry, Isoenzymes chemistry
- Abstract
The arylamine N-acetyltransferases (NATs) NAT1 and NAT2 are responsible for the biotransformation of many arylamine and hydroxylamine xenobiotics. It has been proposed that NATs may act through a cysteine-linked acetyl-enzyme intermediate in a general base catalysis involving a highly conserved arginine residue such as Arg64. To investigate this possibility, we used site-directed mutagenesis and expression of recombinant human NAT1 and NAT2 in Escherichia coli. Sequence comparison with NATs from other species indicated that Arg9 and Arg64 are the only invariant basic residues. Either mutation of the presumed catalytic Cys68 residue or the simultaneous mutation of Arg9 and Arg64 to Ala produced proteins with undetectable enzyme activity. NAT1 or NAT2 singly substituted at Arg9 or Arg64 with Ala, Met, Gln or Lys exhibited unaltered Km values for arylamine acceptor substrates, but a marked loss of activity and stability. Finally, double replacement of Arg9/Arg64 with lysine in NAT1 altered the Km for arylamine substrates (decreased by 8-14-fold) and for acetyl-CoA (elevated 5-fold), and modified the pH-dependence of activity. Thus, through their positively charged side chains, Arg9 and Arg64 seem to contribute to the conformational stability of NAT1 and NAT2 rather than acting as general base catalysts. Our results also support a mechanism in which Arg9 and Arg64 are involved in substrate binding and transition-state stabilization of NAT1.
- Published
- 1997
- Full Text
- View/download PDF
44. N-acetylation genotype and risk of severe reactions to sulphonamides in AIDS patients.
- Author
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Deloménie C, Grant DM, Mathelier-Fusade P, Jacomet C, Leynadier F, Jacqz-Aigrain E, Rozenbaum W, Krishnamoorthy R, and Dupret JM
- Subjects
- Acetylation, Genotype, Humans, Acquired Immunodeficiency Syndrome metabolism, Drug Hypersensitivity, Sulfonamides adverse effects, Sulfonamides metabolism
- Published
- 1994
- Full Text
- View/download PDF
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