Highly individual methylation patterns of alternative glucocorticoid receptor promoters suggest individualized epigenetic regulatory mechanisms

The transcription start sites (TSS) and promoters of many genes are located in upstream CpG islands. Methylation within such islands is known for both imprinted and oncogenes, although poorly studied for other genes, especially those with complex CpG islands containing multiple first exons and promoters. The glucocorticoid receptor (GR) CpG island contains seven alternative first exons and their promoters. Here we show for the five GR promoters activated in PBMCs that methylation patterns are highly variable between individuals. The majority of positions were methylated at levels >25% in at least one donor affecting each promoter and TSS. We also examined the evolutionarily conserved transcription factor binding sites (TFBS) using an improved in silico phylogenetic footprinting technique. The majority of these contain methylatable CpG sites, suggesting that methylation may orchestrates alternative first exon usage, silencing and controlling tissue-specific expression. The heterogeneity observed may reflect epigenetic mechanisms of GR fine tuning, programmed by early life environment and events. With 78% of evolutionarily conserved alternative first exons falling into such complex CpG islands, their internal structure and epigenetic modifications are bound to be biologically important, and may be a common transcriptional control mechanism used throughout many phyla

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Περιέχει την περίληψηΜέσα στα πλαίσια της Κοινωνίας της Πληροφορίας οι βιβλιοθηκονόμοι διεκδικούν, άλλοτε πετυχημένα, άλλοτε όχι και τόσο, ένα πιο ενεργό και απαιτητικό ρόλο, προβάλλοντας το επιχείρημα πώς όσο μεγαλύτερη είναι η παραγωγή πληροφορίας και γνώσης και η παροχή πληροφόρησης τόσο mo απαιτητική είναι και η διαδικασία που αναγκάζεται να ακολουθήσει ο χρήστης ώστε να ικανοποιήσει τις ανάγκες του. Αλλά, η αναγκαιότητα του βιβλιοθηκονόμου (ως επιστήμονα της πληροφόρησης πλέον), του διαμεσολαβητή, δηλαδή, ανάμεσα στην πληροφορία και στον χρήστη, δικαιολογεί απαραιτήτως και την αναγκαιότητα για την ίδια τη Βιβλιοθήκη; Τεκμηριώνεται, δηλαδή, η ύπαρξη αυτού του οργανισμού ως μη κερδοσκοπικού, πολιτιστικού ιδρύματος που εξυπηρετεί όχι μόνο πληροφοριακές αλλά και ψυχαγωγικές, εκπαιδευτικές, κοινωνικοοικονομικές ανάγκες των επισκεπτών του; Και τότε, το έργο του βιβλιοθηκονόμου πώς προδιαγράφεται; Ποιο το περιεχόμενο και ο τρόπος της διαμεσολάβησης; Πρόκειται για σύνθεση ή μηχανιστική διάδοση της πληροφορίας; Αν η Πληροφορία διακινείται, στις μέρες μας κυρίως και πρωτίστως, με ηλεκτρονικά μέσα, τότε το αυτονόητο της ύπαρξης ενός απτού οικοδομήματος που στεγάζει την πληροφορία παύει να ισχύει και είτε πρέπει να καταλυθεί, είτε να εφευρεθεί από την αρχή, προσδίδοντας νέα επίκαιρα χαρακτηριστικά σε ένα αρχαιότατο κατασκεύασμα

Tuberculose abdominale dans une région de faible prévalence tuberculeuse

International audienceObjectiveAbdominal tuberculosis is a rare disease. The clinical and radiological manifestations are non-specific and the diagnosis is difficult. Our objective was to describe the characteristics and treatment of patients presenting with abdominal tuberculosis in a low-incidence country.Patients and methodsWe reviewed the clinical, diagnostic, treatment, and outcome features of patients presenting with abdominal tuberculosis diagnosed by bacteriological and/or histological results and managed in five French university hospitals from January 2000 to December 2009.ResultsWe included 21 patients. The mean diagnostic delay was 13 months. Twelve patients (57%) came from a low-incidence area and only two had a known immunosuppressed condition. Eighteen patients (86%) presented with abdominal symptoms. The main organs involved were the peritoneum (n = 14, 66%), the mesenteric lymph nodes (n = 13, 62%), and the bowel (n = 7, 33%). Sixteen patients (76%) underwent surgery, including two in an emergency setting. Seventeen patients (81%) received six months or more of anti-tuberculosis treatment. Finally, 16 patients (76%) had a positive outcome.ConclusionNew diagnostic procedures, and especially molecular biology, may help diagnose unusual clinical presentations of tuberculosis. Invasive procedures are frequently necessary to obtain samples but also for the treatment of digestive involvement.ObjectifsLa tuberculose abdominale est rare, le tableau clinico-radiologique aspécifique et le diagnostic difficile. Notre objectif était de décrire les caractéristiques et la prise en charge thérapeutique de patients présentant une tuberculose abdominale dans un pays où la prévalence de cette infection est faible.Patients et méthodesNous avons recueilli les données cliniques, diagnostiques, thérapeutiques et le suivi des patients atteints de tuberculose abdominale, diagnostiquée sur des arguments bactériologiques et/ou anatomopathologiques, et pris en charge dans cinq hôpitaux universitaires français de janvier 2000 à décembre 2009.RésultatsVingt et un patients ont été inclus. Le délai diagnostique moyen était de 13 mois. Douze patients (57 %) étaient originaires d’une zone de faible prévalence de la tuberculose et seulement deux patients étaient immunodéprimés. Dix-huit patients (86 %) présentaient des symptômes abdominaux. Les principaux organes atteints étaient le péritoine (n = 14, 66 %), les ganglions mésentériques (n = 13, 62 %) et le tube digestif (n = 7, 33 %). Une intervention chirurgicale a été nécessaire chez 16 patients (76 %), en urgence pour deux d’entre eux. Dix-sept patients (81 %) ont reçu au moins six mois de quadri-thérapie antituberculeuse. Au final, 16 patients (76 %) présentaient une évolution favorable.ConclusionLes nouvelles techniques diagnostiques, notamment la biologie moléculaire, peuvent être utiles pour le diagnostic des formes cliniques inhabituelles de tuberculose. Des explorations invasives sont souvent nécessaires pour obtenir des prélèvements, mais aussi pour la prise en charge thérapeutique de l’atteinte digestive

Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells

CRISPR/Cas9-derived RNA-guided nucleases (RGNs) are DNA targeting systems, which are rapidly being harnessed for gene regulation and gene editing purposes in model organisms and cell lines. As bona fide gene delivery vehicles, viral vectors may be particularly fit to broaden the applicability of RGNs to other cell types including dividing and quiescent primary cells. Here, the suitability of adenoviral vectors (AdVs) for delivering RGN components into various cell types is investigated. We demonstrate that AdVs, namely second-generation fiber-modified AdVs encoding Cas9 or single guide RNA (gRNA) molecules addressing the Cas9 nuclease to the AAVS1 “safe harbor” locus or to a recombinant model allele can be produced to high-titers (up to 20 × 10(10) transducing units/ml). Importantly, AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed cells yields rates of targeted mutagenesis similar to or approaching those achieved by isogenic AdVs encoding TALENs targeting the same AAVS1 chromosomal region. RGN-induced gene disruption frequencies in the various cell types ranged from 18% to 65%. We conclude that AdVs constitute a valuable platform for introducing RGNs into human somatic cells regardless of their transformation status. This approach should aid investigating the potential and limitations of RGNs in numerous experimental settings

Efficacy and safety of one anastomosis gastric bypass versus Roux-en-Y gastric bypass for obesity (YOMEGA): a multicentre, randomised, open-label, non-inferiority trial

International audienceBACKGROUND: One anastomosis gastric bypass (OAGB) is increasingly used in the treatment of morbid obesity. However, the efficacy and safety outcomes of this procedure remain debated. We report the results of a randomised trial (YOMEGA) comparing the outcomes of OAGB versus standard Roux-en-Y gastric bypass (RYGB). METHODS: This prospective, multicentre, randomised non-inferiority trial, was held in nine obesity centres in France. Patients were eligible for inclusion if their body-mass index (BMI) was 40 kg/m(2) or higher, or 35 kg/m(2) or higher with the presence of at least one comorbidity (type 2 diabetes, high blood pressure, obstructive sleep apnoea, dyslipidaemia, or arthritis), and were aged 18-65 years. Key exclusion criteria were a history of oesophagitis, Barrett's oesophagus, severe gastro-oesophageal reflux disease resistant to proton-pump inhibitors, and previous bariatric surgery. Participants were randomly assigned (1:1) to OAGB or RYGB, stratified by centre with blocks of variable size; the study was open-label, with no masking required. RYGB consisted of a 150 cm alimentary limb and a 50 cm biliary limb and OAGB of a single gastrojejunal anastomosis with a 200 cm biliopancreatic limb. The primary endpoint was percentage excess BMI loss at 2 years. The primary endpoint was assessed in the per-protocol population and safety was assessed in all randomised participants. This study is registered with ClinicalTrials.gov, number NCT02139813, and is now completed. FINDINGS: From May 13, 2014, to March 2, 2016, of 261 patients screened for eligibility, 253 (97%) were randomly assigned to OAGB (n=129) or RYGB (n=124). Five patients did not undergo their assigned surgery, and after undergoing their surgery 14 were excluded from the per-protocol analysis (seven due to pregnancy, two deaths, one withdrawal, and four revisions from OAGB to RYGB) In the per-protocol population (n=117 OAGB, n=117 RYGB), mean age was 43.5 years (SD 10.8), mean BMI was 43.9 kg/m(2) (SD 5.6), 176 (75%) of 234 participants were female, and 58 (27%) of 211 with available data had type 2 diabetes. After 2 years, mean percentage excess BMI loss was -87.9% (SD 23.6) in the OAGB group and -85.8% (SD 23.1) in the RYGB group, confirming non-inferiority of OAGB (mean difference -3.3%, 95% CI -9.1 to 2.6). 66 serious adverse events associated with surgery were reported (24 in the RYGB group vs 42 in the OAGB group; p=0.042), of which nine (21.4%) in the OAGB group were nutritional complications versus none in the RYGB group (p=0.0034). INTERPRETATION: OAGB is not inferior to RYGB regarding weight loss and metabolic improvement at 2 years. Higher incidences of diarrhoea, steatorrhoea, and nutritional adverse events were observed with a 200 cm biliopancreatic limb OAGB, suggesting a malabsorptive effect. FUNDING: French Ministry of Health

Digestive perforations related to endoscopy procedures: a local management charter based on local evidence and experts' opinion

International audienceBackground and study aims  Perforations are a known adverse event of endoscopy procedures; a proposal for appropriate management should be available in each center as recommended by the European Society of Gastrointestinal Endoscopy. The objective of this study was to establish a charter for the management of endoscopic perforations, based on local evidence. Patients and methods  Patients were included if they experienced partial or complete perforation during an endoscopic procedure between 2008 and 2018 (retrospectively until 2016, then prospectively). Perforations (size, location, closure) and management (imagery, antibiotics, surgery) were analyzed. Using these results, a panel of experts was asked to propose a consensual management charter. Results  A total of 105 patients were included. Perforations occurred mainly during therapeutic procedures (91, 86.7%). Of the perforations, 78 (74.3 %) were diagnosed immediately and managed during the procedure; 69 of 78 (88.5 %) were successfully closed. Closures were more effective during therapeutic procedures (60 of 66, 90.9 %) than during diagnostic procedures (9 of 12, 75.0 %, P  = 0.06). Endoscopic closure was effective for 37 of 38 perforations (97.4 %) \textless 0.5 cm, and for 26 of 34 perforations (76.5 %) ≥ 0.5 cm ( P  \textless 0.05). For perforations \textless 0.5 cm, systematic computed tomography (CT) scan, antibiotics, or surgical evaluation did not improve the outcome. Four of 105 deaths (3.8 %) occurred after perforation, one of which was attributable to the perforation itself. Conclusions  Detection and closure of perforations during endoscopic procedure had a better outcome compared to delayed perforations; perforations \textless 0.5 cm had a very good prognosis and CT scan, surgeon evaluation, or antibiotics are probably not necessary when the endoscopic closure is confidently performed. This work led to proposal of a local management charter