Identification and characterization of endonuclein binding proteins: evidence of modulatory effects on signal transduction and chaperone activity

<p>Abstract</p> <p>Background</p> <p>We have previously identified endonuclein as a cell cycle regulated WD-repeat protein that is up-regulated in adenocarcinoma of the pancreas. Now, we aim to investigate its biomedical functions.</p> <p>Results</p> <p>Using the cDNA encoding human endonuclein, we have expressed and purified the recombinant protein from <it>Escherichia coli </it>using metal affinity chromatography. The recombinant protein was immobilized to a column and by affinity chromatography several interacting proteins were purified from several litres of placenta tissue extract. After chromatography the eluted proteins were further separated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry. The interacting proteins were identified as; Tax interaction protein 1 (TIP-1), Aα fibrinogen transcription factor (P16/SSBP1), immunoglobulin heavy chain binding protein (BiP), human ER-associated DNAJ (HEDJ/DNAJB11), endonuclein interaction protein 8 (EIP-8), and pregnancy specific β-1 glycoproteins (PSGs). Surface plasmon resonance analysis and confocal fluorescence microscopy were used to further characterize the interactions.</p> <p>Conclusions</p> <p>Our results demonstrate that endonuclein interacts with several proteins indicating a broad function including signal transduction and chaperone activity.</p

Les discours de la Révolution française : aperçu d'ensemble d'un trajet de recherche (1980-2009)

Le chantier d'études sur les langages de la Révolution française, mis en place dans les années 1970 au sein du Laboratoire de lexicologie politique de l'ENS de Saint-Cloud, a pris son essor dans les années 1980, comme en témoigne la revue Mots. Il s'est amplifié sur une base méthodologique précise, l'analyse de discours et la lexicométrie en corpus. Mais il a pris aussi des directions nouvelles telles que l'histoire de la mémoire discursive et l'histoire des révolutions françaises. Enfin, récemment, de nouvelles ouvertures en histoire conceptuelle du discours d'une part, et en histoire du changement linguistique d'autre part, se sont concrétisées sur la base d'un corpus de textes d'écrivains et journalistes révolutionnaires remarqueurs de la nouvelle langue politique, alors que l'édition de corpus numériques reprend de l'ampleur à l'initiative de chercheurs étrangers

Etat Des Lieux et Facteurs Associés en Matière D’eau, D’hygiène Et D’assainissement Dans la Commune d’Abomey-Calavi Au Bénin

Pour apprécier l’accès à l’eau, l’hygiène et à l’assainissement dans la commune d’Abomey-Calavi, un état des lieux a été réalisé et les facteurs associés en matière d’eau, d’hygiène et d’assainissement ont été étudiés. De mai 2016 à janvier 2017, une étude transversale et analytique a permis d’administrer un questionnaire structuré à 630 individus. SPSS et Epi Info ont servi à analyser les facteurs associés. Les focus groupes ont permis de collecter les données qualitatives au sein d’un arrondissement urbain et rural. Un échantillonnage par commodité a permis de sélectionner, à partir des ménages enquêtés, dix participants par arrondissement. La méthode de l’analyse du contenu a été utilisée pour traiter les données qualitatives. L’analyse des résultats obtenus révèle que 61,9% des ménages utilisent une source d’eau améliorée, 87,3% disposent d’installation d’assainissement améliorée, 58,3% et 4,03% disposant respectivement de toilette à chasse mécanique et de toilettes sèches ont une installation d’hygiène améliorée. Le milieu de résidence (OR : 0,3, IC 95% : [0,61-0,42]) explique l’état de la source d’eau de boisson. Le niveau d’instruction (OR : 1,18 ; IC 95% :[0,61-2,25]) et la source d’eau de boisson (OR, 5,62, IC 95% :[2,66-11,85]) expliquent l’état de&nbsp;l’hygiène. La dépense journalière (OR, 0,22, IC 95% : [0,11-0,42]), le milieu de résidence (OR : 0,05 ; IC 95% : [0,02-0,20]), le niveau d’instruction (OR, 0,27 ; IC 95% : [0,16-0,47]) et les traits caractéristiques du relief (OR, 0,27 ; IC 95% : [0,07-1,14]) expliquent l’état de l’assainissement. Il ressort de tout ce qui précède que l’accès à l’eau, l’hygiène et à l’assainissement est acceptable dans la commune d’Abomey-Calavi malgré les disparités entre milieu rural et urbain. Les facteurs associés sont : le niveau d’instruction, la source d’eau de boisson, le niveau économique, le milieu de résidence et les traits caractéristiques du relief. To appreciate the access to water, hygiene and sanitation in the commune of Abomey-Calavi, the inventory was carried out and the associated factors in terms of water, hygiene and sanitation have been studied. FromMay 2016 to January 2017, a cross-sectional and analytical study was used to administer a structured questionnaire to 630 individuals. SPSS and Epi Info were used to analyze the associated factors. The focus groups were used to&nbsp;collect qualitative data in an urban and rural district. A convenience sample were used to select, fromthe households surveyed, ten participants per district. The content analysis method was used to process the qualitative data. The analysis of the results obtained reveals that 61.9% of households use an improved water source, 87.3% have improved sanitation facilities, 58.3% and 4.03% have respectively a flush toilet and dry toilets have an improved hygiene facility. The place of residence (OR: 0.3, 95% CI: [0, 61-0, 42]) explains the state of the drinking water source. The level of education (OR: 1,18, 95% CI: :[0,61-2,25]) and the source of drinking water (OR, 5,62, 95% CI: [2,66-11,85]) explain the state of hygiene. Daily expenditure (OR, 0, 22, 95% CI: [0,11-0,42]), place of residence (OR: 0.05, 95% CI [0,02-0,20]), level of education (OR, 0.27, 95% CI: [0,16-0,47]) and the characteristics of the relief (OR, 0.27, 95% CI: [0,07-1,14]) explain the state of sanitation. From all the foregoing, it is clear that access to water, hygiene and sanitation is acceptable in the commune of Abomey-Calavi despite the disparities between rural and urban areas. Associated factors are: educational level, drinking water source, economic level, place of residence, and characteristics of relief

Single-Cell Gene Expression Analysis of a Human ESC Model of Pancreatic Endocrine Development Reveals Different Paths to β-Cell Differentiation

The production of insulin-producing β cells from human embryonic stem cells (hESCs) in vitro represents a promising strategy for a cell-based therapy for type 1 diabetes mellitus. To explore the cellular heterogeneity and temporal progression of endocrine progenitors and their progeny, we performed single-cell qPCR on more than 500 cells across several stages of in vitro differentiation of hESCs and compared them with human islets. We reveal distinct subpopulations along the endocrine differentiation path and an early lineage bifurcation toward either polyhormonal cells or β-like cells. We uncover several similarities and differences with mouse development and reveal that cells can take multiple paths to the same differentiation state, a principle that could be relevant to other systems. Notably, activation of the key β-cell transcription factor NKX6.1 can be initiated before or after endocrine commitment. The single-cell temporal resolution we provide can be used to improve the production of functional β cells

Le discours politique n’est pas transparent. Permanence et transformations d’un objet de recherche

Peut-on dire que Mots. Les langages du politique a trente ans ? Rien n’est moins sûr. À l’automne 1980, paraît en effet la première livraison d’une publication qui porte pour titre l’acronyme Mots, dont le développement est « Mots, Ordinateurs, Textes, Sociétés ». Ce n’est que neuf ans plus tard que notre revue prendra son nom actuel. Mais seuls des passionnés du discours feront preuve de tels scrupules onomastiques : notre revue a bien trente ans d’âge ! L’équipe rédactionnelle restait la mê..

Hepatitis C of genotype 2: the role of medical invasive exams.

BACKGROUND AND AIM: Hepatitis C virus genotype 2 is the third in order of frequency in Belgium. The aim of this study was to better define the genotype 2 carriers' epidemiology characteristics. METHODS: In a database comprising 1726 viremic hepatitis C virus patient from the south part of Belgium, the files of 98 genotype 2 carriers were reviewed. RESULTS: There was a strong association between genotype 2 and the mode of transmission. The rate of contamination by invasive medical exams was very high (23%), and statistically different from the one of the others genotypes. Eligibility for antiviral therapies and the rate of sustained viral response were high. CONCLUSION: HCV genotype 2 was highly associated with transmission by invasive medical exams.Peer reviewe

Extensive NEUROG3 occupancy in the human pancreatic endocrine gene regulatory network.

OBJECTIVE: Mice lacking the bHLH transcription factor (TF) Neurog3 do not form pancreatic islet cells, including insulin-secreting beta cells, the absence of which leads to diabetes. In humans, homozygous mutations of NEUROG3 manifest with neonatal or childhood diabetes. Despite this critical role in islet cell development, the precise function of and downstream genetic programs regulated directly by NEUROG3 remain elusive. Therefore, we mapped genome-wide NEUROG3 occupancy in human induced pluripotent stem cell (hiPSC)-derived endocrine progenitors and determined NEUROG3 dependency of associated genes to uncover direct targets. METHODS: We generated a novel hiPSC line (NEUROG3-HA-P2A-Venus) where NEUROG3 is HA-tagged and fused to a self-cleaving fluorescent VENUS reporter. We used the CUT&RUN technique to map NEUROG3 occupancy and epigenetic marks in pancreatic endocrine progenitors (PEP) that were differentiated from this hiPSC line. We integrated NEUROG3 occupancy data with chromatin status and gene expression in PEPs as well as their NEUROG3-dependence. In addition, we investigated whether NEUROG3 binds type 2 diabetes mellitus (T2DM)-associated variants at the PEP stage. RESULTS: CUT&RUN revealed a total of 863 NEUROG3 binding sites assigned to 1263 unique genes. NEUROG3 occupancy was found at promoters as well as at distant cis-regulatory elements that frequently overlapped within PEP active enhancers. De novo motif analyses defined a NEUROG3 consensus binding motif and suggested potential co-regulation of NEUROG3 target genes by FOXA or RFX transcription factors. We found that 22% of the genes downregulated in NEUROG3-/- PEPs, and 10% of genes enriched in NEUROG3-Venus positive endocrine cells were bound by NEUROG3 and thus likely to be directly regulated. NEUROG3 binds to 138 transcription factor genes, some with important roles in islet cell development or function, such as NEUROD1, PAX4, NKX2-2, SOX4, MLXIPL, LMX1B, RFX3, and NEUROG3 itself, and many others with unknown islet function. Unexpectedly, we uncovered that NEUROG3 targets genes critical for insulin secretion in beta cells (e.g., GCK, ABCC8/KCNJ11, CACNA1A, CHGA, SCG2, SLC30A8, and PCSK1). Thus, analysis of NEUROG3 occupancy suggests that the transient expression of NEUROG3 not only promotes islet destiny in uncommitted pancreatic progenitors, but could also initiate endocrine programs essential for beta cell function. Lastly, we identified eight T2DM risk SNPs within NEUROG3-bound regions. CONCLUSION: Mapping NEUROG3 genome occupancy in PEPs uncovered unexpectedly broad, direct control of the endocrine genes, raising novel hypotheses on how this master regulator controls islet and beta cell differentiation

Functional Analysis of Ficolin-3 Mediated Complement Activation

The recognition molecules of the lectin complement pathway are mannose-binding lectin and Ficolin -1, -2 and -3. Recently deficiency of Ficolin-3 was found to be associated with life threatening infections. Thus, we aimed to develop a functional method based on the ELISA platform for evaluating Ficolin-3 mediated complement activation that could be applicable for research and clinical use. Bovine serum albumin (BSA) was acetylated (acBSA) and chosen as a solid phase ligand for Ficolins in microtiter wells. Binding of Ficolins on acBSA was evaluated, as was functional complement activation assessed by C4, C3 and terminal complement complex (TCC) deposition. Serum Ficolin-3 bound to acBSA in a calcium dependent manner, while only minimal binding of Ficolin-2 and no binding of Ficolin-1 were observed. No binding to normal BSA was seen for any of the Ficolins. Serum C4, C3 and TCC deposition on acBSA were dependent only on Ficolin-3 in appropriate serum dilutions. Deposition of down stream complement components correlated highly significantly with the serum concentration of Ficolin-3 but not with Ficolin-2 in healthy donors. To make the assay robust for clinical use a chemical compound was applied to the samples that inhibited interference from the classical pathway due to the presence of anti-BSA antibodies in some sera. We describe a novel functional method for measuring complement activation mediated by Ficolin-3 in human serum up to the formation of TCC. The assay provides the possibility to diagnose functional and genetic defects of Ficolin-3 and down stream components in the lectin complement pathway