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991.
Ludovic Maertens Charlotte Erpicum Benoit Detry Silvia Blacher Bénédicte Lenoir Oriane Carnet Christel Péqueux Didier Cataldo Julie Lecomte Jenny Paupert Agnès Noel 《PloS one》2014,9(9)
It is now well accepted that multipotent Bone-Marrow Mesenchymal Stem Cells (BM-MSC) contribute to cancer progression through several mechanisms including angiogenesis. However, their involvement during the lymphangiogenic process is poorly described. Using BM-MSC isolated from mice of two different backgrounds, we demonstrate a paracrine lymphangiogenic action of BM-MSC both in vivo and in vitro. Co-injection of BM-MSC and tumor cells in mice increased the in vivo tumor growth and intratumoral lymphatic vessel density. In addition, BM-MSC or their conditioned medium stimulated the recruitment of lymphatic vessels in vivo in an ear sponge assay, and ex vivo in the lymphatic ring assay (LRA). In vitro, MSC conditioned medium also increased the proliferation rate and the migration of both primary lymphatic endothelial cells (LEC) and an immortalized lymphatic endothelial cell line. Mechanistically, these pro-lymphangiogenic effects relied on the secretion of Vascular Endothelial Growth Factor (VEGF)-A by BM-MSC that activates VEGF Receptor (VEGFR)-2 pathway on LEC. Indeed, the trapping of VEGF-A in MSC conditioned medium by soluble VEGF Receptors (sVEGFR)-1, -2 or the inhibition of VEGFR-2 activity by a specific inhibitor (ZM 323881) both decreased LEC proliferation, migration and the phosphorylation of their main downstream target ERK1/2. This study provides direct unprecedented evidence for a paracrine lymphangiogenic action of BM-MSC via the production of VEGF-A which acts on LEC VEGFR-2. 相似文献
992.
Roscoe Klinck Angélique Fourrier Philippe Thibault Johanne Toutant Mathieu Durand Elvy Lapointe Marie-Laure Caillet-Boudin Nicolas Sergeant Geneviève Gourdon Giovanni Meola Denis Furling Jack Puymirat Benoit Chabot 《PloS one》2014,9(9)
With the goal of identifying splicing alterations in myotonic dystrophy 1 (DM1) tissues that may yield insights into targets or mechanisms, we have surveyed mis-splicing events in three systems using a RT-PCR screening and validation platform. First, a transgenic mouse model expressing CUG-repeats identified splicing alterations shared with other mouse models of DM1. Second, using cell cultures from human embryonic muscle, we noted that DM1-associated splicing alterations were significantly enriched in cytoskeleton (e.g. SORBS1, TACC2, TTN, ACTN1 and DMD) and channel (e.g. KCND3 and TRPM4) genes. Third, of the splicing alterations occurring in adult DM1 tissues, one produced a dominant negative variant of the splicing regulator RBFOX1. Notably, half of the splicing events controlled by MBNL1 were co-regulated by RBFOX1, and several events in this category were mis-spliced in DM1 tissues. Our results suggest that reduced RBFOX1 activity in DM1 tissues may amplify several of the splicing alterations caused by the deficiency in MBNL1. 相似文献
993.
Fanny Morice‐Picard Eulalie Lasseaux Dorothée Cailley Audrey Gros Jérome Toutain Claudio Plaisant Delphine Simon Stéphane François Brigitte Gilbert‐Dussardier Josseline Kaplan Caroline Rooryck Didier Lacombe Benoit Arveiler 《Pigment cell & melanoma research》2014,27(1):59-71
Oculocutaneous albinism (OCA) is caused by mutations in six different genes, and their molecular diagnosis encompasses the search for point mutations and intragenic rearrangements. Here, we used high‐resolution array‐comparative genome hybridization (CGH) to search for rearrangements across exons, introns and regulatory sequences of four OCA genes: TYR, OCA2, TYRP1, and SLC45A2. We identified a total of ten new deletions in TYR, OCA2, and SLC45A2. A complex rearrangement of OCA2 was found in two unrelated patients. Whole‐genome sequencing showed deletion of a 184‐kb fragment (identical to a deletion previously found in Polish patients), whereby a large portion of the deleted sequence was re‐inserted after severe reshuffling into intron 1 of OCA2. The high‐resolution array‐CGH presented here is a powerful tool to detect gene rearrangements. Finally, we review all known deletions of the OCA1–4 genes reported so far in the literature and show that deletions or duplications account for 5.6% of all mutations identified in the OCA1–4 genes. 相似文献
994.
Agnes Cibiel Nam Nguyen Quang Karine Gombert Benoit Thézé Anikitos Garofalakis Frédéric Ducongé 《PloS one》2014,9(1)
Background
Cell-SELEX is now widely used for the selection of aptamers against cell surface biomarkers. However, despite negative selection steps using mock cells, this method sometimes results in aptamers against undesirable targets that are expressed both on mock and targeted cells. Studying these junk aptamers might be useful for further applications than those originally envisaged.Methodology/Principal Findings
Cell-SELEX was performed to identify aptamers against CHO-K1 cells expressing human Endothelin type B receptor (ETBR). CHO-K1 cells were used for negative selection of aptamers. Several aptamers were identified but no one could discriminate between both cell lines. We decided to study one of these aptamers, named ACE4, and we identified that it binds to the Annexin A2, a protein overexpressed in many cancers. Radioactive binding assays and flow cytometry demonstrated that the aptamer was able to bind several cancer cell lines from different origins, particularly the MCF-7 cells. Fluorescence microscopy revealed it could be completely internalized in cells in 2 hours. Finally, the tumor targeting of the aptamer was evaluated in vivo in nude mice xenograft with MCF-7 cells using fluorescence diffuse optical tomography (fDOT) imaging. Three hours after intravenous injection, the aptamer demonstrated a significantly higher uptake in the tumor compared to a scramble sequence.Conclusions/Significance
Although aptamers could be selected during cell-SELEX against other targets than those initially intended, they represent a potential source of ligands for basic research, diagnoses and therapy. Here, studying such aptamers, we identify one with high affinity for Annexin A2 that could be a promising tool for biomedical application. 相似文献995.
996.
David J. P. Ferguson Amanda E. Balaban Eva-Maria Patzewitz Richard J. Wall Christine S. Hopp Benoit Poulin Asif Mohmmed Pawan Malhotra Alida Coppi Photini Sinnis Rita Tewari 《PloS one》2014,9(12)
The circumsporozoite protein (CSP) is the major surface protein of the sporozoite stage of malaria parasites and has multiple functions as the parasite develops and then migrates from the mosquito midgut to the mammalian liver. The overall structure of CSP is conserved among Plasmodium species, consisting of a species-specific central tandem repeat region flanked by two conserved domains: the NH2-terminus and the thrombospondin repeat (TSR) at the COOH-terminus. Although the central repeat region is an immunodominant B-cell epitope and the basis of the only candidate malaria vaccine in Phase III clinical trials, little is known about its functional role(s). We used the rodent malaria model Plasmodium berghei to investigate the role of the CSP tandem repeat region during sporozoite development. Here we describe two mutant parasite lines, one lacking the tandem repeat region (ΔRep) and the other lacking the NH2-terminus as well as the repeat region (ΔNΔRep). We show that in both mutant lines oocyst formation is unaffected but sporozoite development is defective. 相似文献
997.
Activation of protein kinase C (PKC) is thought to involve translocation to the particulate fraction. The present study demonstrates a membrane-associated, inactive pool of PKC in adult rat ventricular myocytes. Membranes were isolated from stimulated (phorbol 12-myristate 13-acetate (PMA), endothelin-1 (ET-1)) or control myocytes and PKC activity determined in the absence (active PKC) or presence (total PKC) of PMA. An inactive, PMA-responsive, pool of PKC was detected. In intact myocytes, PMA or ET-1 induced a translocation of PKC epsilon from the cytosol into the particulate fraction. In contrast, ET-1 decreased both total and active PKC in the membranes: this decrease was associated with a loss of PKC epsilon immunoreactivity. PMA increased the amount of membrane-associated, inactive PKC. Our results demonstrate the presence of a membrane-associated pool of PKC in cardiac myocytes that is differentially modulated by ET-1 or PMA. 相似文献
998.
Ambroise J Giard J Gala JL Macq B 《IEEE/ACM transactions on computational biology and bioinformatics / IEEE, ACM》2011,8(6):1700-1707
A B-cell epitope is a part of an antigen that is recognized by a specific antibody or B-cell receptor. Detecting the immunogenic region of the antigen is useful in numerous immunodetection and immunotherapeutics applications. The aim of this paper is to find relevant properties to discriminate the location of potential epitopes from the rest of the protein surface. The most relevant properties, identified using two evaluation approaches, are the geometric properties, followed by the conservation score and some chemical properties, such as the proportion of glycine. The selected properties are used in a patch-based epitope localization method including a Single-Layer Perceptron for regression. The output of this Single-Layer Perceptron is used to construct a probability map on the antigen surface. The predictive performances of the method are assessed by computing the AUC using cross validation on two benchmark data sets and by computing the AUC and the precision for a third independent test set. 相似文献
999.
Heppell B Blouin S Dussault AM Mulhbacher J Ennifar E Penedo JC Lafontaine DA 《Nature chemical biology》2011,7(6):384-392
S-adenosylmethionine (SAM) riboswitches are widespread in bacteria, and up to five different SAM riboswitch families have been reported, highlighting the relevance of SAM regulation. On the basis of crystallographic and biochemical data, it has been postulated, but never demonstrated, that ligand recognition by SAM riboswitches involves key conformational changes in the RNA architecture. We show here that the aptamer follows a two-step hierarchical folding selectively induced by metal ions and ligand binding, each of them leading to the formation of one of the two helical stacks observed in the crystal structure. Moreover, we find that the anti-antiterminator P1 stem is rotated along its helical axis upon ligand binding, a mechanistic feature that could be common to other riboswitches. We also show that the nonconserved P4 helical domain is used as an auxiliary element to enhance the ligand-binding affinity. This work provides the first comprehensive characterization, to our knowledge, of a ligand-controlled riboswitch folding pathway. 相似文献
1000.
Genomic analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea 总被引:2,自引:0,他引:2
Amselem J Cuomo CA van Kan JA Viaud M Benito EP Couloux A Coutinho PM de Vries RP Dyer PS Fillinger S Fournier E Gout L Hahn M Kohn L Lapalu N Plummer KM Pradier JM Quévillon E Sharon A Simon A ten Have A Tudzynski B Tudzynski P Wincker P Andrew M Anthouard V Beever RE Beffa R Benoit I Bouzid O Brault B Chen Z Choquer M Collémare J Cotton P Danchin EG Da Silva C Gautier A Giraud C Giraud T Gonzalez C Grossetete S Güldener U Henrissat B Howlett BJ Kodira C Kretschmer M Lappartient A Leroch M 《PLoS genetics》2011,7(8):e1002230
Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38-39 Mb genomes include 11,860-14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared to <1% of B. cinerea. The arsenal of genes associated with necrotrophic processes is similar between the species, including genes involved in plant cell wall degradation and oxalic acid production. Analysis of secondary metabolism gene clusters revealed an expansion in number and diversity of B. cinerea-specific secondary metabolites relative to S. sclerotiorum. The potential diversity in secondary metabolism might be involved in adaptation to specific ecological niches. Comparative genome analysis revealed the basis of differing sexual mating compatibility systems between S. sclerotiorum and B. cinerea. The organization of the mating-type loci differs, and their structures provide evidence for the evolution of heterothallism from homothallism. These data shed light on the evolutionary and mechanistic bases of the genetically complex traits of necrotrophic pathogenicity and sexual mating. This resource should facilitate the functional studies designed to better understand what makes these fungi such successful and persistent pathogens of agronomic crops. 相似文献