Deep-sequencing reveals broad subtype-specific HCV resistance mutations associated with treatment failure

A percentage of hepatitis C virus (HCV)-infected patients fail direct acting antiviral (DAA)-based treatment regimens, often because of drug resistance-associated substitutions (RAS). The aim of this study was to characterize the resistance profile of a large cohort of patients failing DAA-based treatments, and investigate the relationship between HCV subtype and failure, as an aid to optimizing management of these patients. A new, standardized HCV-RAS testing protocol based on deep sequencing was designed and applied to 220 previously subtyped samples from patients failing DAA treatment, collected in 39 Spanish hospitals. The majority had received DAA-based interferon (IFN) a-free regimens; 79% had failed sofosbuvir-containing therapy. Genomic regions encoding the nonstructural protein (NS) 3, NS5A, and NS5B (DAA target regions) were analyzed using subtype-specific primers. Viral subtype distribution was as follows: genotype (G) 1, 62.7%; G3a, 21.4%; G4d, 12.3%; G2, 1.8%; and mixed infections 1.8%. Overall, 88.6% of patients carried at least 1 RAS, and 19% carried RAS at frequencies below 20% in the mutant spectrum. There were no differences in RAS selection between treatments with and without ribavirin. Regardless of the treatment received, each HCV subtype showed specific types of RAS. Of note, no RAS were detected in the target proteins of 18.6% of patients failing treatment, and 30.4% of patients had RAS in proteins that were not targets of the inhibitors they received. HCV patients failing DAA therapy showed a high diversity of RAS. Ribavirin use did not influence the type or number of RAS at failure. The subtype-specific pattern of RAS emergence underscores the importance of accurate HCV subtyping. The frequency of “extra-target” RAS suggests the need for RAS screening in all three DAA target regions

Epigrafia e ordine senatorio, 30 anni dopo, I-II

Raccolta di contributi sull'ordine senatorio tra Repubblica e Tardo Impero, mettendo a frutto i risultati della ricerca epigrafic

Thermographic and tomographic methods for tridimensional characterization of thermal transfer in silica/phenolic composites

International audienceSilica/phenolic composite materials are often used in thermal protection systems (TPS) for atmospheric re-entry. The present work aims to compare two different approaches to assess heat transfer properties of these materials: i) using standard and specific experimental methods, and ii) with the development of 3D thermal transfer multiscale model using 2D (microscopy) and 3D (tomography) images. The latter procedure, based on computations on images, is a two-step change of scale from microscopic scale to mesoscopic scale and then to the macroscopic one. Two silica/phenolic composites with different spatial organizations are studied and their 2 thermal properties are compared. Several experimental methods have been used, including space-resolved diffusivity determinations. Numerical results are compared to experimental ones in terms of transverse and longitudinal thermal conductivities of the composites, and were found to be in good agreement. A discussion is made on the different possible sources of uncertainty for both methods

Ablation Properties And Behavior Of Carbon Fiber Reinforced Carbon Composite (c/c Composite)

This paper presents the initial studies carried out in materials used as thermal protection systems (TPS) in reentry atmospheric vehicles (e.g reusable satellites) and in rocket nozzles. In the experiment, graphite and C/C composite are chosen as the target materials. For macroscopic aspect evaluation of the material degradation, the mass losses are measured against the exposure time by changing the material surface temperature. From the microscopic aspect, the eroded surfaces of materials by reactive air plasma are observed with a scanning electron microscope (SEM).Auweter-Kurtz, M., Kurtz, H.L., Laure, S., Plasma generators for re-entry simulation (1996) Journal of Propulsion and Power, 12 (6)Hankey, W.L., (1988) Re-entry Aerodynamics, , Washington, DC: American Institute of Aeronautics and Astronautics, (AIAA Education Series)Loh, W.H.T., (1968) Re-entry and Planetary Entry Physics and Technology, , New York, NY: Spring-VerlangAuweter-Kurtz, M., Laure, S., Röck, W., Experimental planet entry simulation within a plasma wind tunnel (1994) European Symposium on Aerothermo-dynamics for Space 9, , Vehicles, 2nd, Noordwijk. Proceedings. . . Noordwijk: ESTEC, 1994. ESA SP-367Lochte, W., (1968) Holtgreven Plasma Diagnostics, , North-Holland Publishing Company, Amsterda

A(H1N1)pdm09 hemagglutinin D222G and D222N variants are frequently harbored by patients requiring extracorporeal membrane oxygenation and advanced respiratory assistance for severe A(H1N1)pdm09 infection

Background: In patients with A(H1N1)pdm09 infection, severe lung involvement requiring admission to intensive care units (ICU) has been reported. Mutations at the hemagglutinin (HA) receptor binding site (RBS) have been associated with increased virulence and disease severity, representing a potential marker of critical illness. Objectives: To assess the contribution of HA-RBS variability in critically ill patients, A(H1N1)pdm09 virus from adult patients with severe infection admitted to ICU for extracorporeal membrane oxygenation support (ECMO) during influenza season 2009-2011 in Piemonte (4\ub72 million inhabitants), northwestern Italy, was studied. Patients and methods: We retrospectively analyzed HA-RBS polymorphisms in ICU patients and compared with those from randomly selected inpatients with mild A(H1N1)pdm09 disease and outpatients with influenza from the local surveillance program. Results: By HA-RBS direct sequencing of respiratory specimens, D222G and D222N viral variants were identified in a higher proportion in ICU patients (n\ua0=\ua08/24, 33\ub73%) than in patients with mild disease (n\ua0=\ua02/34, 6%) or in outpatients (n\ua0=\ua00/44) (Fisher's exact test P\ua0<\ua00\ub70001; OR 38\ub75; CI 95% 4\ub7494-329\ub79). Eleven ICU patients died (42%), three of them carrying the D222G variant, which was associated with RBS mutation S183P in two. D222G and D222N mutants were identified in upper and lower respiratory samples. Conclusions: A(H1N1)pdm09 HA substitutions D222G and D222N were harbored in a significantly higher proportion by patients with acute respiratory distress for A(H1N1)pdm09 severe infection requiring ICU admission and ECMO. These data emphasize the importance of monitoring viral evolution for understanding virus-host adaptation aimed at the surveillance of strain circulation and the study of viral correlates of disease severity. \ua9 2013 John Wiley & Sons Ltd