Éditeur(s) : HAL CCSD
Résumé : International audience
STUDY QUESTION: Do freezing and in vitro culture procedures enhance the expression of proteins involved in apoptotic or autophagic pathways in murine pre-pubertal testicular tissue? SUMMARY ANSWER: IVM strongly modified apoptosis- and autophagy-related relative protein levels in mice testicular tissue whereas the impact of cryopreservation procedures was minimal at the end of the culture. WHAT IS KNOWN ALREADY: In vitro spermatogenesis remains a challenging technical issue as it imposes to find a very close balance between survival and death of germ cell natural precursors (i.e. gonocytes and spermatogonia), which will eventually undergo a complete spermatogenesis close to in vivo conditions. The establishment of efficient culture conditions coupled with suitable cryopreservation procedures (e.g. controlled slow freezing [CSF] and solid surface vitrification [SSV]) of pre-pubertal testicular tissue is a crucial step in the fields of fertility preservation and restoration to improve the spermatic yield obtained in vitro. STUDY DESIGN, SIZE, DURATION: Here, we study cryopreservation procedures (i.e. CSF or SSV) and the impact of culture media compositions. A first set of 66 mouse pre-pubertal testes were directly cultured during 30, 36, 38 and 60 days (D) from 2.5 to 6.5-day-old CD-1 mice to evaluate the impact of time-aspect of culture and to endorse the reverse phase protein microarrays (RPPM) technique as an adapted experimental tool for the field of in vitro spermatogenesis. Ninety others fresh, slow-frozen and vitrified pre-pubertal testes were cultured during 30 days for the principal study to evaluate the impact of cryopreservation procedures before and after culture. Thirty-four testes dissected from 2.5, 6.5, 36.5, 40.5, 42.5 and 62.5 days postpartum (dpp) mice, corresponding to the time frames of spermatogenesis orchestrated in vitro, were used as in vivo controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: After in vitro culture, testicular tissue samples originated from 2.5 or 6.5-day-old CD-1 male mice were analyzed using RPPM. This targeted proteomic technique allowed us to assess the expression level of 29 apoptosis- and autophagy-related factors by normalizing blank-corrected signal values. In addition, morphological analyses (e.g. HES, PAS, TRA98 and CREM) and DNA fragmentation in intra-tubular cells (i.e. terminal deoxynucleotidyl transferase dUTP nick end labeling; TUNEL) were assessed for the distinct experimental conditions tested as well as for in vivo control mouse testes. MAIN RESULTS AND THE ROLE OF CHANCE: A validation of the RPPM procedure in the field of in vitro spermatogenesis was completed with assay and array robustness before a principal study concerning the evaluation of the impact of in vitro culture and cryopreservation procedures. The proportion of elongated spermatids and the total cell number per seminiferous tubule tended to be very different between the in vivo and in vitro conditions (P < 0.05), suggesting the presence of a beneficial regulation on the first spermatogenesis wave by intrinsic apoptosis (Caspase₉) and autophagy (Atg5) factors (P < 0.0003 and r2 = 0.74). Concerning the impact of culture media compositions, a basic medium (BM) composed of αMEM plus 10% KnockOut™ serum replacement and gentamicin supplemented with retinol (Rol) and vitamin E (Vit. E) was selected as the best culture medium for fresh 6.5 dpp tissue cultured during 30D with 27.7 ± 8.10% of seminiferous tubules containing elongated spermatids. Concerning the impact of cryopreservation procedures, SSV did not have any impact on the morphological parameters evaluated after culture in comparison to fresh tissue (FT) controls. The proportion of tubules with elongated spermatids on testicular explants cultured with BMRol+Vit. E was not different between SSV (6.6 ± 1.6%) and CSF (5.3 ± 1.9%); however, round spermatids were observed more frequently for SSV (19 ± 6.2%) than CSF (3.3 ± 1.9%, P = 0.0317). Even if the proportion of TUNEL-positive cells for BMRol+Vit. E was higher at D30 after SSV (4.12 ± 0.26%) than CSF (1.86 ± 0.12%, P = 0.0022) and FT (2.69 ± 0.33%, P = 0.0108), the DNA damages observed at the end of the culture (i.e. D30) were similar to respective 6.5 dpp controls. In addition, the relative protein level expression ratio of an apoptotic factor, the phosphorylated FADD on Fas, was reduced by 64-fold in vitrified testes cultured with BMRol+Vit. E. Furthermore, we found in this study that the StemPro®-34 SFM culture medium supplemented with growth factors (e.g. EGF, bFGF, GDNF and LIF) prevented the differentiation of spermatogonial stem cells in favor of a significant proliferation with a better architectural pattern than in vivo 6.5 dpp controls with an increase of seminiferous tubules area for FT (P = 0.0357) and CSF (P = 0.0317). LIMITATIONS REASONS FOR CAUTION: Despite our promising results, the evaluation of apoptotic- and autophagic-related proteins was studied for a limited amount of proteins and on global testicular tissue. WIDER IMPLICATIONS OF THE FINDINGS: The data presented herein will help to improve apoptotic and autophagic understanding during the first spermatogenic wave. Moreover, our findings illustrate for the first time that, using finely-tuned experimental conditions, a testicular in vitro culture combined with proteomic technologies may significantly facilitate the study of cryopreservation procedures and in vitro culture evaluations. This study may also contribute to improve work on testicular tissues from pre-pubertal and adolescent cancer survivors. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by a Ph.D. grant from the Rouen Normandie Université and a financial support from 'la Ligue nationale contre le cancer' (both awarded to L.D.), funding from Institute for Research and Innovation in Biomedicine (IRIB), Agence de la Biomédecine, and co-supported by European Union and Région Normandie. Europe gets involved in Normandie with European Regional Development Fund (ERDF). The authors declare that there is no conflict of interest.
Molecular Human Reproduction

hal-01647081
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01647081
DOI : 10.1093/molehr/gax054
PUBMED : 29040674

Éditeur(s) : Elsevier Science Bv
Résumé : There has been growing interest in the development of mud crab aquaculture in New Caledonia. However, for this to become established at a commercial level, a cost-effective formulated feed based on internationally-available ingredients needs to be developed. We have evaluated the optimal dietary protein content for juvenile crabs, Scylla serrata (Forskål, 1775), using a series of diets with a protein content ranging from 27 to 49% and soy protein concentrate (SPC) as the main protein source. For this purpose, 54 individually housed crabs were allocated to five dietary treatments (n = 10 or 11). The crabs were fed ad libitum, for 81 days with the allocated diets. The apparent digestibilities of dry mater, crude protein and energy were high (96.2-97.3%), irrespective of the diet. The voluntary feed intake (VFI) of crabs widely varied from 46 to 220 g kg- 1of fresh initial body weight per week (iBW week- 1) whatever the diet. However, SPC intake and protein intake increased significantly with dietary protein content up to the diet with 40% crude protein, but did not increase further with diets containing 44% and 49% crude protein. The cumulative molts were strongly affected by the VFI levels or energy intake and also, to a lesser extent, by the levels of SPC or protein in diets. Two phases in tissue gain were observed after ecdysis: an initial deposition phase lasting around 30 days followed by a plateau which lasted until the next molt. The daily tissue growth was 16.5% of dry body weight (dry BW) one day after ecdysis and dramatically decreased to 3.6% of dry BW over the first 10 days, then decreased more slowly to the minimum value of 1.3% of dry BW over the next 70 days. During the course of experiment, the best growth (tissue growth and molt frequency) and the best feed efficiency (FCR, PER, retention of proteins and lipids) were obtained with crabs fed on the diet with 40% crude protein. This result was confirmed by a bioenergetic study which showed significantly higher allocation of the energy intake for growth (RE) of crabs fed on diet 40% crude protein. Finally, under our experimental conditions, 1 kg of juvenile crabs required 6.5 ± 1.1 g of protein per day. This level was obtained with the diet SPC-42 that contained 40% of protein of which almost three quarters were derived from SPC. Two hypotheses are proposed to explain the negative effect of high level of SPC or protein on growth and feed efficiency for crabs fed on in diets containing 52% and 60% SPC.
Aquaculture (0044-8486) (Elsevier Science Bv), 2014-10 , Vol. 434 , P. 499-509

Droits : 2014 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/00211/32196/30630.pdf
http://archimer.ifremer.fr/doc/00211/32196/

Éditeur(s) : Inter-Research
Résumé : The pathogenicity of Vibrio penaeicida Strains KH-1 and AM101, their culture-free supernatant (CFS), and their protein fraction obtained by 40% of ammonium sulfate precipitation (PFs40) were assessed in experimental challenges against juvenile Litopenaeus vannamei. Live Vibrio cells, CFS, and PFs40 from the AM101 strain produced a significantly higher mortality (p < 0.05) compared to the KH-1 strain. Toxicity and median lethal doses (LD50) of Fast Protein Liquid Chromatography (FPLC) products were evaluated on L. vannamei. The first FPLC fraction sample (A) from PFs40 of the AM101 strain displayed LD50 values of 1.68 and 5.61 mu g protein ind.(-1), respectively. The second FPLC process from Fraction A showed a peak (A1) also with toxic effects to shrimp. PFs40, Fraction A, and Peak At showed a 38.5 kDa molecular band (SDS-PAGE), with activity on a gelatin protease zymogram. The lethal effect of PFs40 and Fraction A was inhibited by Proteinase K, CuCl2, E-64, and heat (60 and 100 degrees C) treatments, but was not inhibited by EDTA-Na-2, aprotinin, and soy trypsin treatments. These results and the zymogram inhibition test suggest the presence of a cysteine protease-like proteinaceous exotoxin as a dominant protease, secreted by V. penaeicida Strain AM101.
Diseases of aquatic organisms (0177-5103) (Inter-Research), 2005-11 , Vol. 67 , N. 3 , P. 201-207

Droits : Inter-Research 2005
http://archimer.ifremer.fr/doc/2005/publication-3618.pdf
DOI:10.3354/dao067201
http://archimer.ifremer.fr/doc/00000/3618/

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
STUDY QUESTION: Can protein biomarkers of the male genital tract be identified in human seminal plasma? SUMMARY ANSWER: We identified potential biomarkers for each of the organs participating in the secretions of the human seminal plasma. WHAT IS KNOWN ALREADY: The seminal plasma fulfills critical functions for fertility by providing spermatozoa with a protective milieu, promoting their final maturation and modulating the immune responsiveness of the female reproductive tract. It is also considered to be a promising source of biomarkers of male infertility and/or pathologies of the male genital tract. STUDY DESIGN, SIZE, DURATION: This study combines proteomic analyses of normal seminal plasma together with transcriptomic gene expression profiling of human healthy tissues. MATERIALS, SETTING, METHODS: Non-liquefied seminal plasma proteins from a healthy donor were prefractionated using two sequential Proteominer™ libraries. Eight subproteome fractions were collected, trypsin digested and subjected to three successive mass spectrometry analyses for peptide characterization. The list of identified proteins was compared with and merged with other available data sets of the human seminal plasma proteome. The expression of corresponding genes was then investigated using tissue transcriptome profiles to determine where, along the male reproductive tract, these proteins were produced. Finally, tissue specificity of a selected subset of biomarker candidates was validated on human tissues. MAIN RESULTS AND THE ROLE OF CHANCE: We first performed a proteomic analysis of the human seminal plasma and identified 699 proteins. By comparing our protein list with other previous proteomic data sets, we found that 2545 unique proteins have been described so far in the human seminal plasma. We then profiled their expression at the gene level and identified 83 testis, 42 epididymis, 7 seminal vesicle and 17 prostate candidate protein markers. For a subset of testis-specific candidates, i.e. TKTL1, LDHC and PGK2, we further validated their germ cell expression and demonstrated that such markers could distinguish between semen from fertile and infertile men. LIMITATIONS, REASONS FOR CAUTION: While some of the markers we identified are well-known tissue-specific products, further dedicated studies to validate the biomarker status of new candidates will be required. Additionally, whether or not the abundance of these proteins is indeed decreased in some specific pathological situations remains to be determined. WIDER IMPLICATIONS OF THE FINDINGS: Using an integrative genomics approach, we identified biomarker candidates for each of the organs participating in the seminal plasma production. In this study, we essentially focused on germ cell markers and their potential application for the diagnosis of male infertility. Other types of markers also deserve a focused attention given their potential predictive value for various reproductive disorders, notably for prostate cancers. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the Proteomics Core Facility at Biogenouest and was funded by Conseil Régional de Bretagne, IBiSA and Agence de la Biomédecine grants. The authors declare that there exists a competing financial interest in this work that is related to a patent application on the use of identified germ cell-specific proteins in an antibody-based assay (Fertichip™) to predict the successful testicular biopsy outcomes in human non-obstructive azoospermia.
ISSN: 0268-1161

inserm-00814146
http://www.hal.inserm.fr/inserm-00814146
DOI : 10.1093/humrep/des360
PUBMED : 23024119

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Glioblastoma (GB), the most frequent primary tumor of the central nervous system, remains one of the most lethal human cancers despite intensive researches. Current paradigm in the study of GB has been focused on inter-patient variability and on trying to isolate new classification elements or prognostic factors. Here, using ICPL, a technique for protein relative quantification by mass spectrometry, we investigated protein expression between the four regions of GB on clinically relevant biopsies from 5 patients. We identified 584 non-redundant proteins and 31 proteins were found to be up-regulated in the tumor region compared to the peri-tumoral brain tissue, among which, 24 proteins belong to an interaction network linked to 4 biological processes. The core of this network is mainly constituted of interactions between beta-actin (ACTB) with heat shock proteins (HSP90AA1, HSPA8) and 14-3-3 proteins (YWHAZ, YWHAG, YWHAB). A cluster of three isoforms of the sodium pump α-subunit (ATP1A1, ATP1A2, ATP1A3) was also identified outside this network. The differential expression observed for ACTB and 14-3-3γ was further validated by western blot and/or immunohistochemistry. Our study confirms the identity of previously proposed molecular targets, highlights several functional processes altered in GB such as energy metabolism and synaptic transmission and could thus provide added value to new therapeutic trails.
ISSN: 1874-3919

hal-00877756
https://hal.archives-ouvertes.fr/hal-00877756
DOI : 10.1016/j.jprot.2012.04.034
PUBMED : 22575386

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Pyrethroids are widely-used chemical insecticides, to which humans are commonly exposed, and known to alter functional expression of drug metabolizing enzymes. Limited data have additionally suggested that drug transporters, that constitute key-actors of the drug detoxification system, may also be targeted by pyrethroids. The present study was therefore designed to analyze the potential regulatory effects of these pesticides towards activities of main ATP-binding cassette (ABC) and solute carrier (SLC) drug transporters, using transporter-overexpressing cells. The pyrethroids allethrin and tetramethrin were found to inhibit various ABC and SLC drug transporters, including multidrug resistance-associated protein (MRP) 2, breast cancer resistance protein (BCRP), organic anion transporter polypeptide (OATP) 1B1, organic anion transporter (OAT) 3, multidrug and toxin extrusion transporter (MATE) 1, organic cation transporter (OCT) 1 and OCT2, with IC50 values however ranging from 2.6 muM (OCT1 inhibition by allethrin) to 77.6 muM (OAT3 inhibition by tetramethrin) and thus much higher than pyrethroid concentrations (in the nM range) reached in environmentally pyrethroid-exposed humans. By contrast, allethrin and tetramethrin cis-stimulated OATP2B1 activity and failed to alter activities of OATP1B3, OAT1 and MATE2-K, whereas P-glycoprotein activity was additionally moderately inhibited. Twelve other pyrethoids used at 100 muM did not block activities of the various investigated transporters, or only moderately inhibited some of them (inhibition by less than 50%). In silico analysis of structure-activity relationships next revealed that molecular parameters, including molecular weight and lipophilicity, are associated with transporter inhibition by allethrin/tetramethrin and successfully predicted transporter inhibition by the pyrethroids imiprothrin and prallethrin. Taken together, these data fully demonstrated that two pyrethoids, i.e., allethrin and tetramethrin, can act as regulators of the activity of various ABC and SLC drug transporters, but only when used at high and non-relevant concentrations, making unlikely any contribution of these transporter activity alterations to pyrethroid toxicity in environmentally exposed humans.
ISSN: 1932-6203

hal-01477206
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477206
DOI : 10.1371/journal.pone.0169480
PUBMED : 28099443

Éditeur(s) : Université des Antilles Études caribéennes
Résumé : Durant des siècles, les tortues marines ont procuré une source importante de nourriture, notamment de protéines, pour les populations côtières ainsi qu’une source de revenus. Aujourd’hui, elles sont une composante importante de plusieurs cultures à travers le monde et revêtent une signification symbolique importante pour certaines populations insulaires, notamment dans les Petites Antilles.La raréfaction des populations de tortues de mer octroie à ces dernières un certain intérêt touristique, par exemple dans le cadre d’expériences écotouristiques. Cette espèce emblématique peut d’une certaine manière participer à la protection des océans et des récifs coralliens. Dans ce nouveau contexte, leur valeur économique et environnementale est reconsidérée.
During centuries the marine turtles were considered an important resource for food and protein and for the coastal populations and were also a source of revenue .Today they are a major composition of many cultures around the world and have a very symbolical signification for some island populations in the lesser Antilles.The rarefactions of those turtles give them a certain interest for tourists for example and for ecotourism experiences. This symbolical aspect can participate in the protection of oceans and coral reefs this new context their value economical and environmental is re- considered.
Caraïbes

Droits : info:eu-repo/semantics/openAccess
urn:doi:10.4000/etudescaribeennes.6656
http://journals.openedition.org/etudescaribeennes/6656

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Background: The impact of hepatitis B virus (HBV) genomic variability on the measurement of HBsAg level has been poorly evaluated. Objective: This study was designed to compare the performance of all the available assays measuring HBsAg level in this setting. Study design: A large selection of wild type HBV genotypes (n = 184) and HBsAg strains harboring mutations in the S gene (n = 81) from clinical samples was studied with three HBsAg quantification assays: Architect HBsAg (Abbott), LiaisonXL Murex HBsAg Quant (DiaSorin) and the Elecsys HBsAgII (Roche). Results: The overall percentage of positive results was 99.2% for Abbott, 98.9% for DiaSorin and 98.1% for Roche. Abbott and Roche assays provided an excellent concordance in HBsAg quantification (global mean bias of -0.006 logIU/mL). By contrast, DiaSorin underestimated HBsAg level with values 0.112 logIU/ml and 0.103 logIU/ml lower than Abbott and Roche, respectively. By contrast, DiaSorin slightly over quantified gtC(2.5% over the expected value) while Abbott provided values 6.2% lower than expected and 16.2% lower than what observed for the other genotypes. HBsAg quantitative assays were influenced by HBs protein substitutions irrespective to the genotype but no specific protein pattern that would particularly impair the quantification by one technique has been identified. However, Roche seemed to be particularly impacted by substitutions at 145 residue: 75% of under quantified samples carried a substituted 145 residue. Conclusion: This head-to-head comparison indicates a good correlation between all current systems used to quantify HBsAg but clearly shows an influence of both the genotype and the presence of "a" determinant variants in the absolute quantification of HBsAg. While these discrepancies may not translate into major clinical consequence, they may explain an absence of detection of weak concentration of HBsAg on some systems. (C) 2017 Elsevier B.V. All rights reserved.
ISSN: 1386-6532

hal-01588395
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01588395
DOI : 10.1016/j.jcv.2017.02.001
PUBMED : 28189936

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium, the obligate intracellular bacterium that causes heartwater. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia’s OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LCMALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp. The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins.We identified proteins potentially involved in E. ruminantium athogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium.
ISSN: 1932-6203

hal-01203402
https://hal.univ-antilles.fr/hal-01203402
https://hal.univ-antilles.fr/hal-01203402/document
https://hal.univ-antilles.fr/hal-01203402/file/2015_Moumene_PLOSONE-1.pdf
DOI : 10.1371/journal.pone.0116758

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Patients with sickle cell anemia (SCA) have usually lower diastolic, systolic and mean blood pressure (BP) than the general population. However, BP values ≥120/70 mmHg considerably increase the risk for acute and chronic complications in SCA. The aim of this study was to identify biological factors associated with relative hypertension in adults with SCA. We compared the hematological, lipid and hemolytic profiles, as well as blood viscosity, between SCA patients with normal BP (<120/70 mmHg, n = 54) and those with relative hypertension (BP≥120/70 mmHg, n = 43). Our results demonstrated that male gender (OR: 3.49; 95%CI 1.20 to 10.16, p<0.05), triglycerides (OR: 9.19; 95% CI 2.29 to 36.95, p<0.01), blood viscosity (OR: 1.35; 95% CI 1.01 to 1.81, p<0.05) and body mass index (OR: 1.37; 95% CI 1.14 to 1.64, p<0.01) were independent risks factors for relative hypertension in SCA. No association was found between the BP status and the positive history of painful vaso-occlusive crisis or acute chest syndrome. An association between triglycerides level and the occurrence of these two major acute complications was detected. Our study suggests that male gender, increased triglycerides level, BMI and blood viscosity could increase the risk for developing relative hypertension in SCA. In addition, our results support a role of moderately elevated triglycerides in the pathophysiology of vaso-occlusive events.
ISSN: 1932-6203

inserm-00838991
http://www.hal.inserm.fr/inserm-00838991
http://www.hal.inserm.fr/inserm-00838991/document
http://www.hal.inserm.fr/inserm-00838991/file/Lamarre_et_al._2013_PloS_ONE_.pdf
DOI : 10.1371/journal.pone.0066004

Éditeur(s) : HAL CCSD
Résumé : International audience
Gevokizumab is a potent anti-interleukin (IL)-1 beta neutralizing monoclonal antibody (mAb), which may be used for treating inflammatory or autoimmune diseases. The present study was designed to characterize the potential effects of this mAb towards well-established IL-1 beta-mediated repression of hepatic drug detoxifying proteins, like cytochrome P450 (CYP) 3A4 and drug transporters. Primary cultured human hepatocytes were exposed to various concentrations of IL-1 beta in the absence or presence of gevokizumab (5 A mu g/mL); mRNA expression and activity of CYP3A4 and transporters were next determined. Gevokizumab was found to down-modulate, but not abolish, the repression of CYP3A4 and drug transporter mRNAs caused by IL-1 beta in human hepatocytes, through shifting up IL-1 beta half maximal inhibitory concentration (IC50) values by factors ranging from 6.8 to 10.4. The mAb concomitantly shifted IL-1 beta IC50 values towards CYP3A4 activity from 22.0 pg/mL (in the absence of gevokizumab) to 796 pg/mL (in the presence of gevokizumab) and counteracted the decrease of organic anion-transporting polypeptide activity occurring in response to 50 pg/mL IL-1 beta, but not that occurring at higher IL-1 beta concentration (1000 pg/mL). Gevokizumab attenuates, but not abolishes, IL-1 beta-mediated functional repression of CYP3A4 and drug transporters in human hepatocytes, which agrees with the fact that the mAb is considered as a modulator and not a blocker of IL-1 beta signaling. This attenuation of IL-1 beta-mediated down-regulation of hepatic detoxifying proteins by gevokizumab may have to be evaluated in terms of potential therapeutic protein drug-drug interactions when considering future development and therapeutic uses of this IL-1 beta neutralizing mAb.
ISSN: 0378-7966

hal-01616062
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01616062
DOI : 10.1007/s13318-017-0406-1

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
Motivation: Gene fusion is an important evolutionary process. It can yield valuable information to infer the interactions and functions of proteins. Fused genes have been identified as non-transitive patterns of similarity in triplets of genes. To be computationally tractable, this approach usually imposes an a priori distinction between a dataset in which fused genes are searched for, and a dataset that may have provided genetic material for fusion. This reduces the `genetic space' in which fusion can be discovered, as only a subset of triplets of genes is investigated. Moreover, this approach may have a high-false-positive rate, and it does not identify gene families descending from a common fusion event. Results: We represent similarities between sequences as a network. This leads to an efficient formulation of previous methods of fused gene identification, which we implemented in the Python program FusedTriplets. Furthermore, we propose a new characterization of families of fused genes, as clique minimal separators of the sequence similarity network. This well-studied graph topology provides a robust and fast method of detection, well suited for automatic analyses of big datasets. We implemented this method in the C++ program MosaicFinder, which additionally uses local alignments to discard false-positive candidates and indicates potential fusion points. The grouping into families will help distinguish sequencing or prediction errors from real biological fusions, and it will yield additional insight into the function and history of fused genes.
ISSN: 1367-4803

hal-01544795
https://hal.archives-ouvertes.fr/hal-01544795
DOI : 10.1093/bioinformatics/btt049

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
Background: The objective of the study was to determine the incidence of transaminase elevation during dengue, and its predictive factors. Methods: In 2013, a longitudinal study was performed using data from all cases of dengue seen in Cayenne Hospital. Cox proportional modeling was used. Signs of major transaminase elevation were defined as an increase in aspartate amino transferase (AST) or alanine amino transferase (ALT) concentration over 10 times the normal value (10N). Results: There were 1574 patients and 13 249 person-days of follow-up. The incidence rate for signs of trans-aminase elevation (10N) was 0.55 per 100 person-days. Six patients had major transaminase elevation with AST.1000 units (0.43 per 1000 patient-days), and 73 patients (4.6%) developed transaminase elevation with AST .10N. The variables independently associated with major transaminase elevation were hyponatremia, low platelets, dehydration, hematocrit increase, food intolerance, positive nonstructural protein 1 (NS1), age over 15 years and the notion of paracetamol intake. Conclusions: Although very frequent, the incidence of major transaminase elevation was lower than reported elsewhere perhaps because of good access to care, or of the particular serotype causing this epidemic. The patients with transaminase elevation tended to be older, more severe and taking paracetamol.
ISSN: 0035-9203

inserm-01475594
http://www.hal.inserm.fr/inserm-01475594
http://www.hal.inserm.fr/inserm-01475594/document
http://www.hal.inserm.fr/inserm-01475594/file/2016%2C%20Djossou%20-%20Incidence%20and%20predictive%20factors%20of%20transaminase.pdf
DOI : 10.1093/trstmh/trv117

Résumé : Dans les régions tropicales. Il existe une multitude de systèmes alternatifs avec une grande diversité de pratiques. Ces systèmes sont souvent basés sur une utilisation optimale des ressources animales et végétales locales présentes sur l'exploitation. L'objectif de ce travail est de mieux connaître certaines de ces ressources tropicales disponibles et pouvant potentiellement être intéressantes à utiliser en alimentation porcine dans le cadre de ces systèmes alternatifs de type polyculture - élevage. Les résultats montrent que la capacité d'Ingestion des feuilles de manioc et de patate douce est la même quel que soit la forme de présentation (feuilles fraîches ou en farine). La capacité d'ingestion des feuilles de madère est plus élevée lorsqu'elles sont distribuées en farine (95 vs.40SgIj), inversement, les feuilles fraîches d'érythrine sont mieux ingérées (246 vs. 488 g/j). Les résultats sur la valeur nutritionnelle des feuillages tropicaux montrent qu'ils ont une faible densité énergétique comprise entre 6.5 et 8.2 MJ/kg de MS. La détermination du profil en acides aminés des feuilles montrent que les acides aminés des feuilles de madère et de patate douce sont plus digestibles que ceux des feuilles d'érythrine et de manioc.Au final les teneurs en lysine digestible sont de 5,3 g/kg MS pour la patate et 6,3 g/kg MS pour le madère. En conclusion, l'ingestion, ainsi que la teneur énergétique et protéique des feuilles d'une feuille à l'autre varie en fonction de sa teneur en fibres mais également en fonction de leur concentration en tanins. En pratique, les feuilles de patate douce et de madère sont les plus intéressantes pour la réalisation de ration.
In the troples, there are a multitude of alternative systems with wide variety of practices. These systems are often based on an optimal use of local plantand animal on the farm. The objective of this work is to better understand some of these tropical resources avaibable and can potentially be interesting to use in pig feed in the context of these mixed farming system.The results show that the feed intake capacity of cassava and sweet potato leaves is the same whatever the processing form (fresh leaves or meal).The intake capacity of cocoyam leaves18 higher when distributed Into meal (95 vs.408g1d). In opposite the Erythrina fresh leaves are best Ingested (246 va, 488 g 1dl. Results on the nutrltlonal value of tropical foliage show they have low energy density between 6.5 and 8.2 MJ1kg DM. The determlnation of amino acid profile show that the amina acids of cocoyam and sweet potato leaves are more dlgestible than the erythrina and cassava leaves. Ultlmately digestible lysine contents were 5.3 g/kg DM for potato and 6.3 g / kg DM for the cocoyam. ln conclusion, ingestion, and energy and protein content of leaves from one sheet to another depends on its fiber content but aIso according to their tannin. In practice,the leaves of sweet potato and cocoyam are the most Interesting for the realization of diet

http://www.theses.fr/2011AGUY0398/document

Éditeur(s) : HAL CCSD Wiley-VCH Verlag
Résumé : International audience
Proteomic profiling plays a decisive role in the elucidation of molecular signatures representative of a specific clinical context. MuStem cell-based therapy represents a promising approach for clinical applications to cure Duchenne Muscular Dystrophy (DMD). To expand our previous studies collected in the clinically relevant DMD animal model, we decided to investigate the skeletal muscle proteome four months after systemic delivery of allogenic MuStem cells. Quantitative proteomics with isotope-coded protein labelling (ICPL) was used to compile quantitative changes in the protein expression profiles of muscle in transplanted Golden Retriever Muscular Dystrophy (GRMD) dogs as compared to GRMD dogs. A total of 492 proteins were quantified, including 25 that were overrepresented and 46 that were underrepresented after MuStem cell transplantation. Interestingly, this study demonstrates that somatic stem cell therapy impacts on the structural integrity of the muscle fascicle by acting on fibres and its connections with the extracellular matrix. We also show that cell infusion promotes protective mechanisms against oxidative stress and favours the initial phase of muscle repair. This study allows us to identify putative candidates for tissue markers that might be of great value in objectively exploring the clinical benefits resulting from our cell-based therapy for DMD.
ISSN: 1615-9853

Droits : info:eu-repo/semantics/OpenAccess
hal-01332030
https://hal.archives-ouvertes.fr/hal-01332030
https://hal.archives-ouvertes.fr/hal-01332030/document
https://hal.archives-ouvertes.fr/hal-01332030/file/Quantitative%20proteome%20profiling%20of%20dystrophic%20dog%20skeletal%20muscle_accepted.pdf
DOI : 10.1002/pmic.201600002

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Diesel exhaust particles (DEPs) are common environmental air pollutants primarily affecting the lung. DEPs or chemicals adsorbed on DEPs also exert extra-pulmonary effects, including alteration of hepatic drug detoxifying enzyme expression. The present study was designed to determine whether organic DEP extract (DEPe) may target hepatic drug transporters that contribute in a major way to drug detoxification. Using primary human hepatocytes and transporter-overexpressing cells, DEPe was first shown to strongly inhibit activities of the sinusoidal solute carrier (SLC) uptake transporters organic anion-transporting polypeptides (OATP) 1B1, 1B3 and 2B1 and of the canalicular ATP-binding cassette (ABC) efflux pump multidrug resistance-associated protein 2, with IC50 values ranging from approximately 1 to 20 μg/mL and relevant to environmental exposure situations. By contrast, 25 μg/mL DEPe failed to alter activities of the SLC transporter organic cation transporter (OCT) 1 and of the ABC efflux pumps P-glycoprotein and bile salt export pump (BSEP), whereas it only moderately inhibited those of sodium taurocholate co-transporting polypeptide and of breast cancer resistance protein (BCRP). Treatment by 25 μg/mL DEPe was next demonstrated to induce expression of BCRP at both mRNA and protein level in cultured human hepatic cells, whereas it concomitantly repressed mRNA expression of various transporters, including OATP1B3, OATP2B1, OCT1 and BSEP. Such changes in transporter expression were found to be highly correlated to those caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a reference activator of the aryl hydrocarbon receptor (AhR) pathway. This suggests that DEPe, which is enriched in known ligands of AhR like polycyclic aromatic hydrocarbons, alters drug transporter expression via activation of the AhR cascade. Taken together, these data established human hepatic transporters as targets of organic chemicals containing in DEPs, which may contribute to their systemic effects through impairing hepatic transport of endogenous compound or drug substrates of these transporters.
ISSN: 1932-6203

Droits : http://creativecommons.org/licenses/by/
hal-01138535
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01138535
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01138535/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01138535/file/Regulation%20of%20human%20hepatic%20drug.pdf
DOI : 10.1371/journal.pone.0121232

Éditeur(s) : HAL CCSD
Résumé : International audience
The Chromosome-Centric Human Proteome Project (C-HPP) aims to identify "missing" proteins in the neXtProt knowledgebase. We present an in-depth proteomics analysis of the human sperm proteome to identify testis-enriched missing proteins. Using protein extraction procedures and LC-MS/MS analysis, we detected 235 proteins (PE2-PE4) for which no previous evidence of protein expression was annotated. Through LC-MS/MS and LC-PRM analysis, data mining, and immunohistochemistry, we confirmed the expression of 206 missing proteins (PE2-PE4) in line with current HPP guidelines (version 2.0). Parallel reaction monitoring acquisition and sythetic heavy labeled peptides targeted 36 ≪one-hit wonder≫ candidates selected based on prior peptide spectrum match assessment. 24 were validated with additional predicted and specifically targeted peptides. Evidence was found for 16 more missing proteins using immunohistochemistry on human testis sections. The expression pattern for some of these proteins was specific to the testis, and they could possibly be valuable markers with fertility assessment applications. Strong evidence was also found of four "uncertain" proteins (PE5); their status should be re-examined. We show how using a range of sample preparation techniques combined with MS-based analysis, expert knowledge, and complementary antibody-based techniques can produce data of interest to the community. All MS/MS data are available via ProteomeXchange under identifier PXD003947. In addition to contributing to the C-HPP, we hope these data will stimulate continued exploration of the sperm proteome. © 2016 American Chemical Society.
Journal of Proteome Research

hal-01406074
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01406074
DOI : 10.1021/acs.jproteome.6b00400
PUBMED : 27444420

Éditeur(s) : AgroParisTech, ABIES
Résumé : Aquaculture is one of the main supply of protein in feed for mankind, notably in Asia. It increases and had exceeded the level of production of world fisheries, which is decreasing. According to sites and rearing methods, aquaculture contributes to add value but also in some cases to impact negatively the aquatic ecosystems. In the coastal area, a rich and productive zone, but also a place of conflicts, this activity presents a high potential for development if it takes into account the rules of sustainability. The conditions of this sustainability may be cleared with appropriate methods of foresight analysis applied for each development project. The analysis of the professional experience of D. Lacroix for 33 years in aquaculture is structured in three steps. Firstly, the work is to check the scientific value of the research conducted under his responsibility in various contexts and for different responsibilities. This study refers to the requirements of a PhD. In the second part of the work, the results of few major projects (French West Indies, French Guyana….) are analysed with the support of the theory of translation (Callon et al) which allows to build a network of actors and to project it into the context. This way of study includes a special attention to the aspects of time planning, legitimacy and acceptability. Finally, the author examines how this knowledge gained from experience could help building a method for foresight analysis aiming at a shorter planning, a better timing and less means for future projects. To achieve this goal the author integrates the vision of the economist R. Passet which structures any project in an ecosystemic framework, including criteria of sustainability. The mix of these two approaches (Callon & Passet) entails in a new tool for foresight analysis, called Aster, which originality is to offer the possibility to test various scenarios and related consequences to help decision -makers. This new method of foresight analysis, is then tested on the well known case study of the development of prawn Macrobrachium rosenbergii in French Guyana. The test shows to be successful as it delivers precise and arguable recommendations about research and development for all stakeholders.
L'aquaculture est l'une des composantes principales de l'apport en protéines dans le monde, surtout en Asie. Elle croît, alors que la pêche, qu'elle a dépassée en tonnage comme en valeur, décline. Selon les écosystèmes et les méthodes employées, elle peut contribuer à valoriser, mais aussi à dégrader les milieux aquatiques. Dans la zone côtière, espace riche et productif au plan biologique mais convoité, l'aquaculture conserve un potentiel de développement si sa mise en œuvre intègre les principes de la durabilité. Celle-ci peut être éclairée par la prospective avec des méthodes d'analyse adaptées à chaque projet. L'analyse de l'expérience de 33 ans en aquaculture de D. Lacroix est faite en trois temps. D'abord, il s'agit de vérifier la qualité de ses travaux de recherche scientifique menés dans des contextes et avec des responsabilités variés en suivant la grille d'analyse de référence d'un doctorat. Ensuite, l'analyse consiste à prendre un recul critique sur les bilans de réalisations majeures (Antilles, Guyane…) en mobilisant l'approche de M. Callon dont la théorie de la traduction permet de construire le réseau des acteurs et de le projeter dans la problématique. Cette grille d'analyse est complétée par l'étude des aspects de temporalité, légitimité et d'acceptabilité pour chaque réalisation. Ce travail sur diverses situations d'intervention permet de préparer une réflexion sur la proposition méthodologique qui suit et son adaptation à des conditions variables. Enfin, les connaissances acquises sont synthétisées afin de construire une méthode de prospective qui permettrait de gagner en temps, en efficacité et en moyens sur des projets à venir. Pour cela, l'analyse s'appuie sur la vision de R. Passet, qui structure les éléments de tout projet dans la hiérarchie des composantes de la durabilité. Le système dynamique qui en résulte peut être alors croisé avec divers scénarios afin d'en simuler les conséquences et donc d‟éclairer les choix. Cette méthode, dite Aster, est testée sur le cas « modèle » et bien documenté de l'élevage de la crevette Macrobrachium rosenbergii en Guyane française. Elle permet d'aboutir à des recommandations précises et argumentées en matière de recherche comme de développement pour tout décideur impliqué.

Droits : 2010 The author, AgroParisTech, ABIES
http://archimer.ifremer.fr/doc/00032/14349/11633.pdf
http://archimer.ifremer.fr/doc/00032/14349/

Éditeur(s) : HAL CCSD Institute of Electrical and Electronics Engineers
Résumé : International audience
: A new set-up for exposure of human cells in vitro at 37°C to pulse-modulated 300 MHz and 500 MHz signals of future magnetic resonance imaging (MRI) systems is designed, built up, and characterized. Two dipole antennas, specifically designed for ultra high field MRI, are used as radiating structures. The electromagnetic (EM) field distribution inside the incubator containing the cells is computed, and it is shown to be in a good agreement with measurements. The electric field at the cell level is quantified numerically. Local, 1g average, and averaged over the culture medium volume SAR are provided along with the standard deviation values for each well. Temperature increments are measured inside the culture medium during the exposure using an optical fiber thermometer. Then we identify the pulse parameters corresponding to the thermal threshold of 1°C, usually considered as a threshold for thermally-induced biological effects. For these parameters, the induction of heat shock proteins is assessed to biologically verify a potential thermal response of cells. The data demonstrate that, under the considered experimental conditions, exposure to pulsemodulated radiations emulating typical ultra high field MRI signals, corresponding to temperature increments below 1°C, does not trigger any heat shock response in human brain cells.
ISSN: 0018-9294

hal-00874329
https://hal.archives-ouvertes.fr/hal-00874329
DOI : 10.1109/TBME.2013.2270371
PUBMED : 23799680

Éditeur(s) : HAL CCSD
Résumé : International audience
Human hepatoma cells may represent a valuable alternative to the use of human hepatocytes for studying hepatic drug transporters, which is now a regulatory issue during drug development. In the present work, we have characterized hepatic drug transporter expression, activity and regulation in human hepatoma HuH-7 cells, in order to determine the potential relevance of these cells for drug transport assays. HuH-7 cells displayed notable multidrug resistance-associated protein (MRP) activity, presumed to reflect expression of various hepatic MRPs, including MRP2. By contrast, they failed to display functional activities of the uptake transporters sodium taurocholate co-transporting polypeptide (NTCP), organic anion-transporting polypeptides (OATPs) and organic cation transporter 1 (OCT1), and of the canalicular transporters P-glycoprotein and breast cancer resistance protein (BCRP). Concomitantly, mRNA expressions of various sinusoidal and canalicular hepatic drug transporters were not detected (NTCP, OATP1B1, organic anion transporter 2 (OAT2), OCT1 and bile salt export pump) or were found to be lower (OATP1B3, OATP2B1, multidrug and toxin extrusion protein 1, BCRP and MRP3) in hepatoma HuH-7 cells than those found in human hepatocytes, whereas other transporters such as OAT7, MRP4 and MRP5 were up-regulated. HuH-7 cells additionally exhibited farnesoid X receptor (FXR)- and nuclear factor erythroid
Pharmaceutics

hal-01477210
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477210
DOI : 10.3390/pharmaceutics9010003
PUBMED : 28036031