Optimisation of recombinant production of active human cardiac SERCA2a ATPase

Methods for recombinant production of eukaryotic membrane proteins, yielding sufficient quantity and quality of protein for structural biology, remain a challenge. We describe here, expression and purification optimisation of the human SERCA2a cardiac isoform of Ca2+ translocating ATPase, using Saccharomyces cerevisiae as the heterologous expression system of choice. Two different expression vectors were utilised, allowing expression of C-terminal fusion proteins with a biotinylation domain or a GFP- His8 tag. Solubilised membrane fractions containing the protein of interest were purified onto Streptavidin-Sepharose, Ni-NTA or Talon resin, depending on the fusion tag present. Biotinylated protein was detected using specific antibody directed against SERCA2 and, advantageously, GFP-His8 fusion protein was easily traced during the purification steps using in-gel fluorescence. Importantly, talon resin affinity purification proved more specific than Ni-NTA resin for the GFP-His8 tagged protein, providing better separation of oligomers present, during size exclusion chromatography. The optimised method for expression and purification of human cardiac SERCA2a reported herein, yields purified protein (> 90%) that displays a calcium-dependent thapsigargin-sensitive activity and is suitable for further biophysical, structural and physiological studies. This work provides support for the use of Saccharomyces cerevisiae as a suitable expression system for recombinant production of multi-domain eukaryotic membrane proteins

1st ESMO Consensus Conference in lung cancer; Lugano 2010: Small-cell lung cancer

The 1st ESMO Consensus Conference on lung cancer was held in Lugano, Switzerland on 21st and 22nd May 2010 with the participation of a multidisciplinary panel of leading professionals in pathology and molecular diagnostics and medical, surgical and radiation oncology. Before the conference, the expert panel prepared clinically relevant questions concerning five areas as follows: early and locally advanced non-small-cell lung cancer (NSCLC), first-line metastatic NSCLC, second-/third-line NSCLC, NSCLC pathology and molecular testing, and small-cell lung cancer (SCLC) to be addressed through discussion at the Consensus Conference. All relevant scientific literature for each question was reviewed in advance. During the Consensus Conference, the panel developed recommendations for each specific question. The consensus agreement in SCLC is reported in this article. The recommendations detailed here are based on an expert consensus after careful review of published data. All participants have approved this final updat

Entanglement Entropy, decoherence, and quantum phase transition of a dissipative two-level system

The concept of entanglement entropy appears in multiple contexts, from black hole physics to quantum information theory, where it measures the entanglement of quantum states. We investigate the entanglement entropy in a simple model, the spin-boson model, which describes a qubit (two-level system) interacting with a collection of harmonic oscillators that models the environment responsible for decoherence and dissipation. The entanglement entropy allows to make a precise unification between entanglement of the spin with its environment, decoherence, and quantum phase transitions. We derive exact analytical results which are confirmed by Numerical Renormalization Group arguments both for an ohmic and a subohmic bosonic bath. Those demonstrate that the entanglement entropy obeys universal scalings. We make comparisons with entanglement properties in the quantum Ising model and in the Dicke model. We also emphasize the possibility of measuring this entanglement entropy using charge qubits subject to electromagnetic noise; such measurements would provide an empirical proof of the existence of entanglement entropy.Comment: 38 pages, 8 figures, related to cond-mat/0612095 and arXiv:0705.0957; final version to appear in Annals of Physic

Deflated GMRES for Systems with Multiple Shifts and Multiple Right-Hand Sides

We consider solution of multiply shifted systems of nonsymmetric linear equations, possibly also with multiple right-hand sides. First, for a single right-hand side, the matrix is shifted by several multiples of the identity. Such problems arise in a number of applications, including lattice quantum chromodynamics where the matrices are complex and non-Hermitian. Some Krylov iterative methods such as GMRES and BiCGStab have been used to solve multiply shifted systems for about the cost of solving just one system. Restarted GMRES can be improved by deflating eigenvalues for matrices that have a few small eigenvalues. We show that a particular deflated method, GMRES-DR, can be applied to multiply shifted systems. In quantum chromodynamics, it is common to have multiple right-hand sides with multiple shifts for each right-hand side. We develop a method that efficiently solves the multiple right-hand sides by using a deflated version of GMRES and yet keeps costs for all of the multiply shifted systems close to those for one shift. An example is given showing this can be extremely effective with a quantum chromodynamics matrix.Comment: 19 pages, 9 figure

Sterile neutrino production via active-sterile oscillations: the quantum Zeno effect

We study several aspects of the kinetic approach to sterile neutrino production via active-sterile mixing. We obtain the neutrino propagator in the medium including self-energy corrections up to $\mathcal{O}(G^2_F)$, from which we extract the dispersion relations and damping rates of the propagating modes. The dispersion relations are the usual ones in terms of the index of refraction in the medium, and the damping rates are $\Gamma_1(k) = \Gamma_{aa}(k) \cos^2\theta_m(k); \Gamma_2(k) = \Gamma_{aa}(k) \sin^2\theta_m(k)$ where $\Gamma_{aa}(k)\propto G^2_F k T^4$ is the active neutrino scattering rate and $\theta_m(k)$ is the mixing angle in the medium. We provide a generalization of the transition probability in the \emph{medium from expectation values in the density matrix}: $P_{a\to s}(t) = \frac{\sin^22\theta_m}{4}[e^{-\Gamma_1t} + e^{-\Gamma_2 t}-2e^{-{1/2}(\Gamma_1+\Gamma_2)t} \cos\big(\Delta E t\big)]$ and study the conditions for its quantum Zeno suppression directly in real time. We find the general conditions for quantum Zeno suppression, which for $m_s\sim \textrm{keV}$ sterile neutrinos with $\sin2\theta \lesssim 10^{-3}$ \emph{may only be} fulfilled near an MSW resonance. We discuss the implications for sterile neutrino production and argue that in the early Universe the wide separation of relaxation scales far away from MSW resonances suggests the breakdown of the current kinetic approach.Comment: version to appear in JHE

Variational theory of elastic manifolds with correlated disorder and localization of interacting quantum particles

We apply the gaussian variational method (GVM) to study the equilibrium statistical mechanics of the two related systems: (i) classical elastic manifolds, such as flux lattices, in presence of columnar disorder correlated along the $\tau$ direction (ii) interacting quantum particles in a static random potential. We find localization by disorder, the localized phase being described by a replica symmetry broken solution confined to the mode $\omega=0$. For classical systems we compute the correlation function of relative displacements. In $d=2+1$, in the absence of dislocations, the GVM allows to describes the Bose glass phase. Along the columns the displacements saturate at a length $l_{\perp}$ indicating flux-line localization. Perpendicularly to the columns long range order is destroyed. We find divergent tilt modulus $c_{44}=\infty$ and a $x \sim \tau^{1/2}$ scaling. Quantum systems are studied using the analytic continuation from imaginary to real time $\tau \to i t$. We compute the conductivity and find that it behaves at small frequency as $\sigma(\omega) \approx \omega^2$ in all dimensions ($d < 4$) for which disorder is relevant. We compute the quantum localization length $\xi$. In $d=1$, where the model also describes interacting fermions in a static random potential, we find a delocalization transition and obtain analytically both the low and high frequency behavior of the conductivity for any value of the interaction. We show that the marginality condition appears as the condition to obtain the correct physical behavior. Agreement with renormalization group results is found whenever it can be compared.Comment: 34 pages, REVTeX, no figure

Are MRI-defined fat infiltrations in the multifidus muscles associated with low back pain?

BACKGROUND: Because training of the lumbar muscles is a commonly recommended intervention in low back pain (LBP), it is important to clarify whether lumbar muscle atrophy is related to LBP. Fat infiltration seems to be a late stage of muscular degeneration, and can be measured in a non-invasive manner using magnetic resonance imaging. The purpose of this study was to investigate if fat infiltration in the lumbar multifidus muscles (LMM) is associated with LBP in adults and adolescents. METHODS: In total, 412 adults (40-year-olds) and 442 adolescents (13-year-olds) from the general Danish population participated in this cross-sectional cohort study. People with LBP were identified through questionnaires. Using MRI, fat infiltration of the LMM was visually graded as none, slight or severe. Odds ratios were calculated for both age groups, taking into account sex, body composition and leisure time physical activity for both groups, and physical workload (in adults only) or daily bicycling (in adolescents only). RESULTS: Fat infiltration was noted in 81% of the adults but only 14% of the adolescents. In the adults, severe fat infiltration was strongly associated with ever having had LBP (OR 9.2; 95% CI 2.0–43.2), and with having LBP in the past year (OR 4.1; 1.5–11.2), but there was no such association in adolescents. None of the investigated moderating factors had an obvious effect on the OR in the adults. CONCLUSION: Fat infiltration in the LMM is strongly associated with LBP in adults only. However, it will be necessary to quantify these measurements objectively and to investigate the direction of this link longitudinally in order to determine if the abnormal muscle is the cause of LBP or vice versa

Entanglement entropy of two disjoint blocks in XY chains

We study the Renyi entanglement entropies of two disjoint intervals in XY chains. We exploit the exact solution of the model in terms of free Majorana fermions and we show how to construct the reduced density matrix in the spin variables by taking properly into account the Jordan-Wigner string between the two blocks. From this we can evaluate any Renyi entropy of finite integer order. We study in details critical XX and Ising chains and we show that the asymptotic results for large blocks agree with recent conformal field theory predictions if corrections to the scaling are included in the analysis correctly. We also report results in the gapped phase and after a quantum quench.Comment: 34 pages, 11 figure