Genome-wide study predicts promoter-G4 DNA motifs regulate selective functions in bacteria: radioresistance of D. radiodurans involves G4 DNA-mediated regulation

A remarkable number of guanine-rich sequences with potential to adopt non-canonical secondary structures called G-quadruplexes (or G4 DNA) are found within gene promoters. Despite growing interest, regulatory role of quadruplex DNA motifs in intrinsic cellular function remains poorly understood. Herein, we asked whether occurrence of potential G4 (PG4) DNA in promoters is associated with specific function(s) in bacteria. Using a normalized promoter-PG4-content (PG4P) index we analysed >60 000 promoters in 19 well-annotated species for (a) function class(es) and (b) gene(s) with enriched PG4P. Unexpectedly, PG4-associated functional classes were organism specific, suggesting that PG4 motifs may impart specific function to organisms. As a case study, we analysed radioresistance. Interestingly, unsupervised clustering using PG4P of 21 genes, crucial for radioresistance, grouped three radioresistant microorganisms including Deinococcus radiodurans. Based on these predictions we tested and found that in presence of nanomolar amounts of the intracellular quadruplex-binding ligand N-methyl mesoporphyrin (NMM), radioresistance of D. radiodurans was attenuated by ∼60%. In addition, important components of the RecF recombinational repair pathway recA, recF, recO, recR and recQ genes were found to harbour promoter-PG4 motifs and were also down-regulated in presence of NMM. Together these results provide first evidence that radioresistance may involve G4 DNA-mediated regulation and support the rationale that promoter-PG4s influence selective functions

Regulation of Neutrophil Degranulation and Cytokine Secretion: A Novel Model Approach Based on Linear Fitting

Neutrophils participate in the maintenance of host integrity by releasing various cytotoxic proteins during degranulation. Due to recent advances, a major role has been attributed to neutrophil-derived cytokine secretion in the initiation, exacerbation, and resolution of inflammatory responses. Because the release of neutrophil-derived products orchestrates the action of other immune cells at the infection site and, thus, can contribute to the development of chronic inflammatory diseases, we aimed to investigate in more detail the spatiotemporal regulation of neutrophil-mediated release mechanisms of proinflammatory mediators. Purified human neutrophils were stimulated for different time points with lipopolysaccharide. Cells and supernatants were analyzed by flow cytometry techniques and used to establish secretion profiles of granules and cytokines. To analyze the link between cytokine release and degranulation time series, we propose an original strategy based on linear fitting, which may be used as a guideline, to (i) define the relationship of granule proteins and cytokines secreted to the inflammatory site and (ii) investigate the spatial regulation of neutrophil cytokine release. The model approach presented here aims to predict the correlation between neutrophil-derived cytokine secretion and degranulation and may easily be extrapolated to investigate the relationship between other types of time series of functional processes

Rice Galaxy: An open resource for plant science

Background: Rice molecular genetics, breeding, genetic diversity, and allied research (such as rice-pathogen interaction) have adopted sequencing technologies and high-density genotyping platforms for genome variation analysis and gene discovery. Germplasm collections representing rice diversity, improved varieties, and elite breeding materials are accessible through rice gene banks for use in research and breeding, with many having genome sequences and high-density genotype data available. Combining phenotypic and genotypic information on these accessions enables genome-wide association analysis, which is driving quantitative trait loci discovery and molecular marker development. Comparative sequence analyses across quantitative trait loci regions facilitate the discovery of novel alleles. Analyses involving DNA sequences and large genotyping matrices for thousands of samples, however, pose a challenge to non−computer savvy rice researchers. Findings: The Rice Galaxy resource has shared datasets that include high-density genotypes from the 3,000 Rice Genomes project and sequences with corresponding annotations from 9 published rice genomes. The Rice Galaxy web server and deployment installer includes tools for designing single-nucleotide polymorphism assays, analyzing genome-wide association studies, population diversity, rice−bacterial pathogen diagnostics, and a suite of published genomic prediction methods. A prototype Rice Galaxy compliant to Open Access, Open Data, and Findable, Accessible, Interoperable, and Reproducible principles is also presented. Conclusions: Rice Galaxy is a freely available resource that empowers the plant research community to perform state-of-the-art analyses and utilize publicly available big datasets for both fundamental and applied science

Neutralization profiles of HIV-1 viruses from the VRC01 Antibody Mediated Prevention (AMP) trials

The VRC01 Antibody Mediated Prevention (AMP) efficacy trials conducted between 2016 and 2020 showed for the first time that passively administered broadly neutralizing antibodies (bnAbs) could prevent HIV-1 acquisition against bnAb-sensitive viruses. HIV-1 viruses isolated from AMP participants who acquired infection during the study in the sub-Saharan African (HVTN 703/HPTN 081) and the Americas/European (HVTN 704/HPTN 085) trials represent a panel of currently circulating strains of HIV-1 and offer a unique opportunity to investigate the sensitivity of the virus to broadly neutralizing antibodies (bnAbs) being considered for clinical development. Pseudoviruses were constructed using envelope sequences from 218 individuals. The majority of viruses identified were clade B and C; with clades A, D, F and G and recombinants AC and BF detected at lower frequencies. We tested eight bnAbs in clinical development (VRC01, VRC07-523LS, 3BNC117, CAP256.25, PGDM1400, PGT121, 10–1074 and 10E8v4) for neutralization against all AMP placebo viruses (n = 76). Compared to older clade C viruses (1998–2010), the HVTN703/HPTN081 clade C viruses showed increased resistance to VRC07-523LS and CAP256.25. At a concentration of 1μg/ml (IC80), predictive modeling identified the triple combination of V3/V2-glycan/CD4bs-targeting bnAbs (10-1074/PGDM1400/VRC07-523LS) as the best against clade C viruses and a combination of MPER/V3/CD4bs-targeting bnAbs (10E8v4/10-1074/VRC07-523LS) as the best against clade B viruses, due to low coverage of V2-glycan directed bnAbs against clade B viruses. Overall, the AMP placebo viruses represent a valuable resource for defining the sensitivity of contemporaneous circulating viral strains to bnAbs and highlight the need to update reference panels regularly. Our data also suggests that combining bnAbs in passive immunization trials would improve coverage of global viruses

Agrégation de classifieurs et d'experts pour la recherche d'homologues chez les cytokines à quatre hélices alpha

I was working on a particular gene family : the four helix cytokines. The major purpose of this work was to design a new method to detected still unknown members in the human genome. The first part of my work was to compare SVM classifiers, which is known as the best strategy for homologs research, from the literature. During the second part of my work, i designed automatical experts which deals with information like biological features.The last part of my work consisted in evaluating methods to aggregate classifiers and experts. This strategy achieve better results than the best classifer alone and it can easily be adapted to other gene family.Agrégation de classifieurs et d'experts pour la recherche d'homologues chez les cytokines à quatre hélices alpha , Texte imprimé -- Nicolas Beaume ; sous la direction de Yannick Jacques, Gérard Ramstein L'objectif de ce travail est la mise au point d'une méthode de détection d'homologues de cytokines inconnues. J'ai dans un premier temps évalué plusieurs classifieurs SVM. J'ai ensuite proposé d'ajouter, sous la forme d'experts automatiques, des connaissances spécifiques à la famille étudiée. Enfin, afin de maximiser l'efficacité de leur association, j'ai comparé différentes méthodes d'agrégation. Je propose une méthode performante, basée sur la combinaisons de ces classifieurs et de ces experts, généralisable à d'autres familles de protéines

Agrégation de classi eurs et d'experts pour la recherche d'homologues chez les cytokines à quatre hélices alpha

L'objectif de ce travail est la mise au point d'une méthode de détection d'homologues de cytokines inconnues. J'ai dans un premier temps évalué plusieurs classifieurs SVM. J'ai ensuite proposé d'ajouter, sous la forme d'experts automatiques, des connaissances spécifiques à la famille étudiée. Enfin, afin de maximiser l'efficacité de leur association, j'ai comparé différentes méthodes d'agrégation. Je propose une méthode performante, basée sur la combinaisons de ces classifieurs et de ces experts, généralisable à d'autres familles de protéines.I was working on a particular gene family : the four helix cytokines. The major purpose of this work was to design a new method to detected still unknown members in the human genome. The first part of my work was to compare SVM classifiers, which is known as the best strategy for homologs research, from the literature. During the second part of my work, i designed automatical experts which deals with information like biological features.The last part of my work consisted in evaluating methods to aggregate classifiers and experts. This strategy achieve better results than the best classifer alone and it can easily be adapted to other gene family.NANTES-BU Médecine pharmacie (441092101) / SudocSudocFranceF

Título: Gramática completa.

Texto en español y francés.Suplemento con portadilla y pág. propia

Observations in-situ des mouvements du bain de fusion et des mécanismes de solidification au cours de ligne de fusion TIG

Ce document est un rapport d’étude qui présente les travaux effectués par l'équipe Assemblages Soudés (AS) du LMGC (Univ. Montpellier - CNRS, UMR 5508) dans le cadre du projet ANR Nemesis (Projet ANR-17-CE08-0036 du Programme AAPG 2017). Il s’agit d’une étude expérimentale réalisée principalement au cours du post-doctorat de Nicolas Blanc. Le but de cette étude est de pouvoir observer la solidification et le comportement du bain de soudage in-situ pour deux types d’alliages d’intérêt industriel : l’acier 316L et l’acier 22MnB5, intéressant de façon prioritaire respectivement EDF et Arcelor Mittal, partenaires du projet ANR. Les expériences doivent permettre d’obtenir des informations sur la taille et la forme du bain, sur les écoulements existants en son sein et sur le processus de solidification. Pour cela un dispositif similaire à celui utilisé par Alexis Chiocca [1] est utilisé. Le document est organisé autour de quatre chapitres. Le premier est dédié à un succinct état de l’art afin de rappeler le contexte scientifique général et l’utilité des observations réalisées dans la compréhension des phénomènes de solidifications pendant le soudage. Le deuxième chapitre est consacré à la présentation du dispositif expérimental spécifiquement conçu pour ce type de mesures et à la présentation des méthodologies d’étude. Les deux derniers chapitres sont consacrés à la présentation des résultats obtenus respectivement pour l’acier inoxydable 316L et les alliages 22MnB5

Comparison of a healthy miRNome with melanoma patient miRNomes: are microRNAs suitable serum biomarkers for cancer?

MiRNAs are increasingly recognized as biomarkers for the diagnosis of cancers where they are profiled from tumor tissue (intracellular miRNAs) or serum/plasma samples (extracellular miRNAs). To improve detection of reliable biomarkers from blood samples, we first compiled a healthy reference miRNome and established a well-controlled analysis pipeline allowing for standardized quantification of circulating miRNAs. Using whole miRNome and custom qPCR arrays, miRNA expression profiles were analyzed in 126 serum, whole blood and tissue samples of healthy volunteers and melanoma patients and in primary melanocyte and keratinocyte cell lines. We found characteristic signatures with excellent prognostic scores only in late stage but not in early stage melanoma patients. Upon comparison of melanoma tissue miRNomes with matching serum samples, several miRNAs were identified to be exclusively tissue-derived (miR-30b-5p, miR-374a-5p and others) while others had higher expression levels in serum (miR-3201 and miR-122-5p). Here we have compiled a healthy and widely applicable miRNome from serum samples and we provide strong evidence that levels of cell-free miRNAs only change significantly at later stages of melanoma progression, which has serious implications for miRNA biomarker studies in cancer

Identification of SOCS2 and SOCS6 as biomarkers in human colorectal cancer.

BACKGROUND: Over the past years, some members of the family of suppressor of cytokine signalling (SOCS) proteins have emerged as potential tumour suppressors. This study aimed at investigating the clinical significance of SOCS proteins in colorectal carcinoma (CRC). METHODS: We integrated publicly available microarray expression data on CRC in humans, analysed the expression pattern of SOCSs and assessed the predictive power of SOCS2 and SOCS6 for diagnostic purposes by generating receiver operating characteristic curves. Using laser microdissected patient material we assessed SOCS expression on RNA and protein levels as well as their methylation status in an independent CRC patient cohort. Finally, we investigated the prognostic value of SOCS2 and SOCS6. RESULTS: The meta-analysis as well as the independent patient cohort analysis reveal a stage-independent downregulation of SOCS2 and SOCS6 and identify both molecules as diagnostic biomarkers for CRC. We demonstrate a different methylation pattern within the SOCS2 promoter between tumour tissue and normal control tissue in 25% of CRC patients. Furthermore, early CRC stage patients with low expression of SOCS2 display significantly shorter disease-free survival. CONCLUSIONS: Our data offers evidence that SOCS2 and SOCS6 levels are reduced in CRC and may serve as diagnostic biomarkers for CRC patients