Suppression of “leaky” nonsense mutations by ribosome readthrough accounts for residual factor IX levels in Haemophilia B patients

INTRODUCTION Nonsense mutations, relatively frequent in Haemophilia (>10%), are considered “null mutations”. However, they have been found also in moderate/mild Haemophiliacs and, as compared with large gene deletions, are associated with lower risk for developing inhibitors. This observation point to the presence of residual expression levels arising from “ribosome readthrough” over nonsense triplets, as we have shown in a very small cohort of Haemophilia B (HB) patients. Drugs inducing readthrough such as aminoglycosides are proposed as potential therapy. AIM To investigate residual levels of factor IX (FIX) produced by an extended panel of HB nonsense mutations. METHODS Expression of nonsense FIX variants in HEK293 cells, evaluation of intracellular and secreted FIX levels (ELISA, Western Blotting, fluorogenic functional assays). RESULTS We investigated nine F9 mutations (R75X, L103X, R162X, R294X, R298X, Y330X, Q370X, R379X, R384X) associated to severe/moderate HB. Appreciable levels of secreted FIX were detected for the R162X, R294X, R298X, Y330X mutants, with a large predominance of truncated forms. Truncated forms for Q370X, R379X and R384X were observed in cell lysates only, which suggests misfolding. Noticeably, the full-length FIX form was appreciable for the R162X, R294X and R298X and indicated the occurrence of spontaneous readthrough, which was significantly increased by the use of aminoglycosides. Moreover, aminoglycosides promoted the synthesis of full-length FIX in the presence of the R75X, Y330X, Q370X, R379X, R384X mutations, not undergoing appreciable spontaneous readthrough. Preliminary functional assays revealed a residual activity for the R162X mutant. CONCLUSIONS These data demonstrate that nonsense mutations can be associated to residual FIX levels through a mechanism of “productive” readthrough. The identification of “leaky” nonsense mutations, and thus of patients with trace FIX levels and potentially “high responders” to readthrough-inducing drugs, might help diagnosis and treatment

Compound effect of EHD and surface roughness in pool boiling and CHF with R-123

This article is a post-print version of the fianl published article which may be accessed at the link below.Saturated pool boiling of R-123 at 1 bar, including the critical heat flux (CHF), was enhanced by modifying the surface characteristics and applying a high intensity electrostatic field, the latter termed electrohydrodynamic (and abbreviated EHD) enhancement. The heat flux was varied from very low values in the natural convection regime up to CHF. Experiments were performed with increasing and decreasing heat flux to study boiling hysteresis without and with EHD. Boiling occurred on the sand blasted surface of a cylindrical copper block with embedded electrical heating elements, with standardized surface parameter Pa = 3.5 μm. The electric field was generated by a potential of 5 kV to 25 kV, applied through a 40 mm diameter circular electrode of ss-304 wire mesh, aperture size 5.1 mm, located at distances of 5 - 60 mm from the surface, with most of the data obtained for 20 mm. The data for the rough surface were compared with earlier data for a smooth surface and indicated a significant increase in the heat transfer rates. EHD produced a further increase in the heat transfer rates, particularly at low heat flux values and near the CHF. Boiling hysteresis was reduced progressively by EHD and eliminated at high field strength.This work was supported by Government of Pakistan under a scholarship programme

Dark sector studies with the PADME experiment

The Positron Annihilation to Dark Matter Experiment (PADME) uses the positron beam of the DAΦNE Beam-Test Facility, at the Laboratori Nazionali di Frascati (LNF) to search for a Dark Photon A′. The search technique studies the missing mass spectrum of singlephoton final states in e+e− → A′γ annihilation in a positron-on-thin-target experiment. This approach facilitates searches for new particles such as long lived Axion-Like-Particles, protophobic X bosons and Dark Higgs. This talk illustrated the scientific program of the experiment and its first physics results. In particular, the measurement of the cross-section of the SM process e+e− →γγ at ps=21 MeV was shown

Search for diphoton events with large missing transverse momentum in 1 fb(-1) of 7 TeV proton-proton collision data with the ATLAS detector ATLAS Collaboration

A search for diphoton events with large missing transverse momentum has been performed using 1.07 fb -1 of proton-proton collision data at s=7 TeV recorded with the ATLAS detector. No excess of events was observed above the Standard Model prediction and 95% Confidence Level (CL) upper limits are set on the production cross section for new physics. The limits depend on each model parameter space and vary as follows: Λ<(22-129) fb in the context of a generalised model of gauge-mediated supersymmetry breaking (GGM) with a bino-like lightest neutralino, σ<(27-91) fb in the context of a minimal model of gauge-mediated supersymmetry breaking (SPS8), and σ<(15-27) fb in the context of a specific model with one universal extra dimension (UED). A 95% CL lower limit of 805 GeV, for bino masses above 50 GeV, is set on the GGM gluino mass. Lower limits of 145 TeV and 1.23 TeV are set on the SPS8 breaking scale Λ and on the UED compactification scale 1/R, respectively. These limits provide the most stringent tests of these models to date. © 2012 CERN

Search for diphoton events with large missing transverse momentum in 1 fb^-1 of 7 TeV proton–proton collision data with the ATLAS detector

A search for diphoton events with large missing transverse momentum has been performed using 1.07 fb−1of proton–proton collision data at √s=7  TeV recorded with the ATLAS detector. No excess of events was observed above the Standard Model prediction and 95% Confidence Level (CL) upper limits are set on the production cross section for new physics. The limits depend on each model parameter space and vary as follows: σ<(22–129) fb in the context of a generalised model of gauge-mediated supersymmetry breaking (GGM) with a bino-like lightest neutralino, σ<(27–91) fb in the context of a minimal model of gauge-mediated supersymmetry breaking (SPS8), and σ< (15–27) fb in the context of a specific model with one universal extra dimension (UED). A 95% CL lower limit of 805 GeV, for bino masses above 50 GeV, is set on the GGM gluino mass. Lower limits of 145 TeV and 1.23 TeV are set on the SPS8 breaking scale ∧ and on the UED compactification scale 1/R, respectively. These limits provide the most stringent tests of these models to date

Search for diphoton events with large missing transverse momentum in 1 fb(-1) of 7 TeV proton-proton collision data with the ATLAS detector ATLAS Collaboration

A search for diphoton events with large missing transverse momentum has been performed using 1.07 fb(-1) of proton-proton collision data at root s = 7 TeV recorded with the ATLAS detector. No excess of events was observed above the Standard Model prediction and 95% Confidence Level (CL) upper limits are set on the production cross section for new physics. The limits depend on each model parameter space and vary as follows: sigma < (22-129) fb in the context of a generalised model of gauge-mediated supersymmetry breaking (GGM) with a bino-like lightest neutralino, sigma < (27-91) fb in the context of a minimal model of gauge-mediated supersymmetry breaking (SPS8), and sigma < (15-27) fb in the context of a specific model with one universal extra dimension (UED). A 95% CL lower limit of 805 GeV, for bino masses above 50 GeV, is set on the GGM gluino mass. Lower limits of 145 TeV and 1.23 TeV are set on the SPS8 breaking scale Lambda and on the UED compactification scale 1/R, respectively. These limits provide the most stringent tests of these models to date. (C) 2012 CERN. Published by Elsevier B.V. All rights reserved

Search for diphoton events with large missing transverse momentum in 1 fb(-1) of 7 TeV proton-proton collision data with the ATLAS detector ATLAS Collaboration

A search for diphoton events with large missing transverse momentum has been performed using 1.07 fb(-1) of proton-proton collision data at root s = 7 TeV recorded with the ATLAS detector. No excess of events was observed above the Standard Model prediction and 95% Confidence Level (CL) upper limits are set on the production cross section for new physics. The limits depend on each model parameter space and vary as follows: sigma < (22-129) fb in the context of a generalised model of gauge-mediated supersymmetry breaking (GGM) with a bino-like lightest neutralino, sigma < (27-91) fb in the context of a minimal model of gauge-mediated supersymmetry breaking (SPS8), and sigma < (15-27) fb in the context of a specific model with one universal extra dimension (UED). A 95% CL lower limit of 805 GeV, for bino masses above 50 GeV, is set on the GGM gluino mass. Lower limits of 145 TeV and 1.23 TeV are set on the SPS8 breaking scale Lambda and on the UED compactification scale 1/R, respectively. These limits provide the most stringent tests of these models to date. (C) 2012 CERN. Published by Elsevier B.V. All rights reserved. RI Sivoklokov, Sergey/D-8150-2012; Li, Xuefei/C-3861-2012; Smirnov, Sergei/F-1014-2011; Gladilin, Leonid/B-5226-2011; Barreiro, Fernando/D-9808-2012; Prokoshin, Fedor/E-2795-2012; Fazio, Salvatore /G-5156-2010; Orlov, Ilya/E-6611-2012; Doyle, Anthony/C-5889-2009; Alexa, Calin/F-6345-2010; Moorhead, Gareth/B-6634-2009; Livan, Michele/D-7531-2012; Takai, Helio/C-3301-2012; Petrucci, Fabrizio/G-8348-2012; Jones, Roger/H-5578-2011; Fabbri, Laura/H-3442-2012; Kurashige, Hisaya/H-4916-2012; Villa, Mauro/C-9883-2009; Delmastro, Marco/I-5599-201