Controlled assembly of SNAP-PNA-fluorophore systems on DNA templates to produce fluorescence resonance energy transfer

The SNAP protein is a widely used self-labeling tag that can be used for tracking protein localization and trafficking in living systems. A model system providing controlled alignment of SNAP-tag units can provide a new way to study clustering of fusion proteins. In this work, fluorescent SNAP-PNA conjugates were controllably assembled on DNA frameworks forming dimers, trimers, and tetramers. Modification of peptide nucleic acid (PNA) with the O6-benzyl guanine (BG) group allowed the generation of site-selective covalent links between PNA and the SNAP protein. The modified BG-PNAs were labeled with fluorescent Atto dyes and subsequently chemo-selectively conjugated to SNAP protein. Efficient assembly into dimer and oligomer forms was verified via size exclusion chromatography (SEC), electrophoresis (SDS-PAGE), and fluorescence spectroscopy. DNA directed assembly of homo- and hetero-dimers of SNAP-PNA constructs induced homo- and hetero-FRET, respectively. Longer DNA scaffolds controllably aligned similar fluorescent SNAP-PNA constructs into higher oligomers exhibiting homo-FRET. The combined SEC and homo-FRET studies indicated the 1:1 and saturated assemblies of SNAP-PNA-fluorophore:DNA formed preferentially in this system. This suggested a kinetic/stoichiometric model of assembly rather than binomially distributed products. These BG-PNA-fluorophore building blocks allow facile introduction of fluorophores and/or assembly directing moieties onto any protein containing SNAP. Template directed assembly of PNA modified SNAP proteins may be used to investigate clustering behavior both with and without fluorescent labels which may find use in the study of assembly processes in cells

Gaviscon® vs. omeprazole in symptomatic treatment of moderate gastroesophageal reflux. a direct comparative randomised trial

<p>Abstract</p> <p>Background</p> <p>Medical management of GERD mainly uses proton pump inhibitors. Alginates also have proven efficacy. The aim of this trial was to compare short-term efficacy of an alginate (Gaviscon^®, 4 × 10 mL/day) and omeprazole (20 mg/day) on GERD symptoms in general practice.</p> <p>Methods</p> <p>A 14-day multicentre randomised double-blind double-dummy non-inferiority trial compared Gaviscon^®(4 × 10 mL/day) and omeprazole (20 mg/day) in patients with 2-6 day heartburn episodes weekly without alarm signals. The primary outcome was the mean time to onset of the first 24-h heartburn-free period after initial dosing. Secondary outcomes were the proportion of patients without heartburn by D7, pain relief by D7, and reduction in pain intensity by D7 and D14.</p> <p>Results</p> <p>278 patients were recruited; 120 were included in the Gaviscon^®group and 121 in the omeprazole group for the per protocol non-inferiority analysis. The mean time to onset of the first 24-h heartburn-free period after initial dosing was 2.0 (± 2.2) days for Gaviscon^®and 2.0 (± 2.3) days for omeprazole (<it>p </it>= 0.93); mean intergroup difference was 0.01 ± 1.55 days (95% CI = -0.41 to 0.43): i.e., less than the lower limit of the 95% CI of -0.5 days predetermined to demonstrate non-inferiority. The mean number of heartburn-free days by D7 was significantly greater in the omeprazole group: 3.7 ± 2.3 days vs. 3.1 ± 2.1 (<it>p </it>= 0.02). On D7, overall quality of pain relief was slightly in favour of omeprazole (<it>p </it>= 0.049). There was no significant difference in the reduction in pain intensity between groups by D7 (<it>p = </it>0.11) or D14 (<it>p = </it>0.08). Tolerance and safety were good and comparable in both groups.</p> <p>Conclusion</p> <p>Gaviscon^®was non-inferior to omeprazole in achieving a 24-h heartburn-free period in moderate episodic heartburn, and is a relevant effective alternative treatment in moderate GERD in primary care.</p> <p>Trial registration</p> <p><a href="http://www.controlled-trials.com/ISRCTN62203233">ISRCTN62203233</a>.</p

A prospective observational study of a cohort of outpatients with an acute medical event and reduced mobility: incidence of symptomatic thromboembolism and description of thromboprophylaxis practices.

OBJECTIVES: The study was performed to determine the incidence of symptomatic venous thromboembolism in outpatients with an acute medical event causing temporary reduced mobility. Risk factors for venous thromboembolism and thromboprophylaxis practices were also studied. DESIGN: This was a prospective, observational, multicentre, cohort study. SETTING: General practitioners randomly selected from a registry of 25,000 active representative doctors in France including eligible outpatients. SUBJECTS: Outpatients aged at least 40 years anticipated to have reduced mobility for at least 48 h due to an acute medical event were eligible. INTERVENTIONS: None required. MAIN OUTCOME MEASURES: Symptomatic deep-vein thrombosis and pulmonary embolism at 3 weeks were the main study end-points. RESULTS: Overall, 16,532 evaluable patients of mean age 71 years were recruited between October 2002 and June 2003 by 2895 doctors. The main acute medical events leading to reduced mobility were infection, acute rheumatism and falls without fracture. The incidence rates (95% confidence interval) of symptomatic deep-vein thrombosis and pulmonary embolism were 1% (0.84-1.14) and 0.20% (0.13-0.27) respectively. Venous insufficiency in legs, cancer, and a personal or family history of venous thromboembolism were independent risk factors for venous thromboembolism. Pharmacological thromboprophylaxis was initiated in 35.0% (n=5782) of the patients. The principal driver of prescription was a personal history of venous thromboembolism. CONCLUSIONS: The risk of symptomatic venous thromboembolism in outpatients with reduced mobility for medical reasons is close to that reported in medical and surgical inpatients. This risk and the potential need for thromboprophylaxis should be taken into account by primary care doctors

La vaccination contre la Covid-19 : le vaccin Comirnaty de Pfizer-BioNtech

Le conseil scientifique du CNGE a rédigé un outil d'aide à la décision pour les médecins généralistes concernant la vaccination contre la COVID-19. Il s'agit d'aider le clinicien à discuter des données scientifiques avec le patient pour l'aider à prendre une décision

Joint Bone Spine

INTRODUCTION: Physical therapy (PT) represents a major approach in musculoskeletal (MSK) pain. This study aimed to assess kinesiophobia, its impact and management, in patients with MSK pain treated by PT. METHODS: A national multicentre, prospective study was conducted in France in patients with MSK pain referred to PT. Kinesiophobia was scored with the Tampa Scale of Kinesiophobia (TSK). Pain, satisfaction, analgesic intake and acceptability were assessed at the initial visit, at the 5th PT session, and at the end of PT. RESULTS: 700 consecutive outpatients with MSK pain, 54.5% female, referred to PT were recruited by 186 GPs: 501 had significant levels of kinesiophobia (TSK score > 40). Patients with kinesiophobia were significantly older, with less physical activity, more pain and less acceptability. Patients from GPs presenting with kinesiophobia had both higher pain and kinesiophobia levels. After 5 PT sessions, global satisfaction was significantly higher in patients without kinesiophobia. A significant increase of PT satisfaction was observed in patients who had been given preventive analgesics before PT sessions, in 25.6% of patients. Independent predictors for specific management of PT-induced pain were: patient's kinesiophobia (OR = 2.02 [1.07-3.82]), current analgesics treatment (OR=2.05 [1.16-3.63]), GP with postgraduate course on pain (OR = 2.65 [1.29-5.43]), GP's independent practice (OR = 1.88 [1.01-3.48]). CONCLUSION: Kinesiophobia is frequent in patients with MSK pain, is associated to GPs' kinesiophobia and decreases satisfaction of Physical Therapy. Preventive analgesic treatment before PT sessions improves patients' satisfaction and should be proposed to improve MSK pain management

Utilisation du fondaparinux 2,5mg pour la prévention des évènements thromboemboliques veineux en médecine générale. ArchiMed Ville

International audienceOBJECTIVE:To evaluate the mean duration of treatment course with fondaparinux 2.5 mg (ARIXTRA(®)) in the setting of ambulatory general medicine, with respect to its indication in thromboprophylaxis for medically ill patients and to describe the population treated.METHODS:Observational, prospective, national, multicenter, pharmaco-epidemiological study, performed in France, at the request of the Transparency Commission (a division of the French Health Regulatory Authority). The general practitioners had to include the first three adult patients, considered as patients at high risk of venous thromboembolic events and immobilized for acute medical illness, treated with initiation of thromboprophylaxis by fondaparinux 2.5 mg.RESULTS:Two hundred and seventeen general practitioners included 840 patients. The mean age of patients was 63.6±18.1 years, and 63% of patients (n=520/831) were females. The real total administration duration of the treatment by fondaparinux 2.5 mg was known for 797 patients and was 15.8±12.4 days on average (range: 1-90 days, median: 10 days). In 40% of patients, the duration ranged from 6 to 14 days [duration consistent with the summary of product characteristics (SmPC)]. Among the 834 patients analyzed, 569 (68%) suffered from at least one acute illness and had at least one risk factor for venous thromboembolism (VTE). The indication did fully comply with the summary of product characteristics of fondaparinux 2.5 mg in 52% of the patients (n=434/834 patients).CONCLUSION:The results of the ArchiMed study support that the thromboprophylaxis treatment with fondaparinux 2.5 mg in ambulatory general medicine, and the associated medical conditions were usually consistent with the SmPC or guidelines. However, a difference was found for the duration and the initial indication, in situations that may be regarded as presenting a risk by the prescriber

Prévention des évènements thromboemboliques veineux par fondaparinux 2,5mg chez des patients hospitalisés pour une affection médicale aiguë. Étude ArchiMed. ArchiMed hôpital étude observationnelle d'usage du fondaparinux 2,5mg en vie réelle ; 718 patients inclus par 107 pharmaciens hospitaliers.

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