Raf-1 kinase associates with Hepatitis C virus NS5A and regulates viral replication

AbstractHepatitis C virus (HCV) is a positive-strand RNA virus that frequently causes persistent infection associated with severe liver disease. HCV nonstructural protein 5A (NS5A) is essential for viral replication. Here, the kinase Raf-1 was identified as a novel cellular binding partner of NS5A, binding to the C-terminal domain of NS5A. Raf-1 colocalizes with NS5A in the HCV replication complex. The interaction of NS5A with Raf-1 results in increased Raf-1 phosphorylation at serine 338. Integrity of Raf-1 is crucial for HCV replication: inhibition of Raf-1 by the small-molecule inhibitor BAY43-9006 or downregulation of Raf-1 by siRNA attenuates viral replication

Demonstration of Gd-GEM detector design for neutron macromolecular crystallography applications

The European Spallation Source (ESS) in Lund, Sweden will become the world's most powerful thermal neutron source. The Macromolecular Diffractometer (NMX) at the ESS requires three 51.2 x 51.2~cm$^{2}$ detectors with reasonable detection efficiency, sub-mm spatial resolution, a narrow point spread function (PSF) and good time resolution. This work presents measurements with the improved version of the NMX detector prototype consisting of a Triple-GEM detector with natural Gd converter and a low material budget readout. The detector was successfully tested at the neutron reactor of the Budapest Neutron Centre (BNC) and at the D16 instrument at the Institut Laue-Langevin (ILL) in Grenoble. The measurements with Cadmium and Gadolinium masks in Budapest demonstrate that the point spread function of the detector lacks long tails that could impede the measurement of diffraction spot intensities. On the D16 instrument at ILL, diffraction spots from Triose phosphate isomerase w/ 2-phosphoglycolate (PGA) inhibitor were measured both in the D16 Helium-3 detector and the Gd-GEM. The comparison between the two detectors show a similar point spread function in both detectors, and the expected efficiency ratio compared to the Helium-3 detector. Both measurements together thus give good indications that the Gd-GEM detector fits the requirements for the NMX instrument at ESS

X-ray imaging with gaseous detectors using the VMM3a and the SRS

The integration of the VMM3a Application-Specific Integrated Circuit (ASIC) into RD51's Scalable Readout System (SRS) provides a versatile tool for the readout of Micro-Pattern Gaseous Detectors (MPGDs). With its self-triggered high-rate readout, its analogue part that allows to get information on the deposited energy in the detector, and its so-called neighbouring-logic that allows to recover information on the charge distribution, this new system has features of particular interest for digital X-ray imaging. In the present article, we want to emphasise the capabilities of VMM3a/SRS by presenting results of X-ray imaging studies. We will highlight the advantages on the energy and the spatial resolution provided by the neighbouring-logic. In the first part, we focus on spatial resolution studies. We show how segmented readout structures introduce a repeating pattern in the distribution of the reconstructed positions (using the centre-of-gravity method) and how this behaviour can be mitigated with the neighbouring-logic. As part of these studies, we explore as well an alternative position reconstruction algorithm. In the second part of the article, we present the energy resolution studies.Peer reviewe

Hepatitis C virus cell-cell transmission and resistance to direct-acting antiviral agents

Hepatitis C virus (HCV) is transmitted between hepatocytes via classical cell entry but also uses direct cell-cell transfer to infect neighboring hepatocytes. Viral cell-cell transmission has been shown to play an important role in viral persistence allowing evasion from neutralizing antibodies. In contrast, the role of HCV cell-cell transmission for antiviral resistance is unknown. Aiming to address this question we investigated the phenotype of HCV strains exhibiting resistance to direct-acting antivirals (DAAs) in state-of-the-art model systems for cell-cell transmission and spread. Using HCV genotype 2 as a model virus, we show that cell-cell transmission is the main route of viral spread of DAA-resistant HCV. Cell-cell transmission of DAA-resistant viruses results in viral persistence and thus hampers viral eradication. We also show that blocking cell-cell transmission using host-targeting entry inhibitors (HTEIs) was highly effective in inhibiting viral dissemination of resistant genotype 2 viruses. Combining HTEIs with DAAs prevented antiviral resistance and led to rapid elimination of the virus in cell culture model. In conclusion, our work provides evidence that cell-cell transmission plays an important role in dissemination and maintenance of resistant variants in cell culture models. Blocking virus cell-cell transmission prevents emergence of drug resistance in persistent viral infection including resistance to HCV DAAs

Development of the scalable readout system for micro-pattern gas detectors and other applications

Developed within RD51 Collaboration for the Development of Micro-Pattern Gas Detectors Technologies, the Scalable Readout System (SRS) is intended as a general purpose multichannel readout solution for a wide range of detector types and detector complexities. The scalable architecture, achieved using multi-Gbps point-to-point links with no buses involved, allows the user to tailor the system size to his needs. The modular topology enables the integration of different front-end ASICs, giving the user the possibility to use the most appropriate front-end for his purpose or to build a heterogeneous experimental apparatus which integrates different front-ends into the same DAQ system. Current applications include LHC upgrade activities, geophysics or homeland security applications as well as detector R&D. The system architecture, development and running experience will be presented, together with future prospects, ATCA implementation options and application possibilities.Martoiu, S.; Muller, H.; Tarazona Martínez, A.; Toledo Alarcón, JF. (2013). Development of the scalable readout system for micro-pattern gas detectors and other applications. Journal of Instrumentation. 8(3):1-11. doi:10.1088/1748-0221/8/03/C03015S1118

Rate-capability of the VMM3a front-end in the RD51 Scalable Readout System

The VMM3a is an Application Specific Integrated Circuit (ASIC), specifically developed for the readout of gaseous detectors. Originally developed within the ATLAS New Small Wheel (NSW) upgrade, it has been successfully integrated into the Scalable Readout System (SRS) of the RD51 collaboration. This allows, to use the VMM3a also in small laboratory set-ups and mid-scale experiments, which make use of Micro-Pattern Gaseous Detectors (MPGDs). As part of the integration of the VMM3a into the SRS, the readout and data transfer scheme was optimised to reach a high rate-capability of the entire readout system and profit from the VMM3a’s high single-channel rate-capability of 3.6 Mhits∕s. The optimisation focused mainly on the handling of the data output stream of the VMM3a, but also on the development of a trigger-logic between the front-end cards and the DAQ computer. In this article, two firmware implementations of the non-ATLAS continuous readout mode are presented, as well as the implementation of the trigger-logic. Afterwards, a short overview on X-ray imaging results is presented, to illustrate the high rate-capability from an application point-of-view.Peer reviewe

Infrastructure for Detector Research and Development towards the International Linear Collider

The EUDET-project was launched to create an infrastructure for developing and testing new and advanced detector technologies to be used at a future linear collider. The aim was to make possible experimentation and analysis of data for institutes, which otherwise could not be realized due to lack of resources. The infrastructure comprised an analysis and software network, and instrumentation infrastructures for tracking detectors as well as for calorimetry.Comment: 54 pages, 48 picture

Neutralizing antibodies and pathogenesis of hepatitis C virus infection.

Hepatitis C virus (HCV) infection is a major cause of chronic liver disease worldwide. The interplay between the virus and host innate and adaptive immune responses determines the outcome of infection. There is increasing evidence that host neutralizing responses play a relevant role in the resulting pathogenesis. Furthermore, viral evasion from host neutralizing antibodies has been revealed to be an important contributor in leading both to viral persistence in acute liver graft infection following liver transplantation, and to chronic viral infection. The development of novel model systems to study HCV entry and neutralization has allowed a detailed understanding of the molecular mechanisms of virus-host interactions during antibody-mediated neutralization. The understanding of these mechanisms will ultimately contribute to the development of novel antiviral preventive strategies for liver graft infection and an urgently needed vaccine. This review summarizes recent concepts of the role of neutralizing antibodies in viral clearance and protection, and highlights consequences of viral escape from neutralizing antibodies in the pathogenesis of HCV infection