NP:P and NP:V Interactions of the Paramyxovirus Simian Virus 5 Examined Using a Novel Protein:Protein Capture Assay

AbstractUsing recombinant proteins extracted from mammalian cells, in a novel protein:protein binding assay, direct interaction of the nucleoprotein (NP) of simian virus 5 with the phosphoprotein (P) and V protein (V) was demonstrated. The amount of NP bound by V was found to be significantly less than that bound by P. Furthermore, preabsorption of NP with P removed the fraction of NP that could be bound by V, but preabsorption of NP with V did not remove all the NP that could be bound by P. These results suggested that V bound a subpopulation of the NP recognised by P. Further analysis revealed that P bound both soluble and homopolymeric forms of NP, while V bound only the soluble form; thus demonstrating that the binding sites on P and V, for soluble NP, are located within the N-terminal domain common to both P and V proteins. A monoclonal antibody, which recognised an epitope in the unique C-terminus of P, blocked the binding of P to polymeric NP but not to soluble NP. These results also suggest that there are two binding sites on NP for P; the site that interacts with the P/V common domain being either hidden or conformationally altered in polymeric NP

Sex differences in endothelial function in porcine coronary arteries: a role for H2O2and gap junctions?

BACKGROUND AND PURPOSE: Cardiovascular risk is higher in men and postmenopausal women compared with premenopausal women. This may be due to sex differences in endothelial function. Here, sex differences in endothelial function of porcine coronary arteries (PCAs) were investigated. EXPERIMENTAL APPROACH: Distal PCAs were studied under myographic conditions and after precontraction with U46619. Concentration-response curves to bradykinin were constructed in the presence of a range of inhibitors. KEY RESULTS: In male and female PCAs, bradykinin produced comparable vasorelaxant responses. Inhibition of NO and prostanoid synthesis produced greater inhibition in males compared with females. Removing H2O2 with PEG-catalase reduced the maximum relaxation in the absence, but not the presence of L-NAME and indomethacin in females, and had no effect in males. Blocking gap junctions with 100 muM carbenoxolone or 18α-glycyrrhetinic acid further inhibited the endothelium-derived hyperpolarization (EDH-mediated response in females but not in males. In female PCAs, the maximum EDH-mediated response was reduced by inhibiting SKCa with apamin and by inhibiting IKCa with TRAM-34, or with both. In male PCAs, at maximum bradykinin concentration, the EDH-mediated response was reduced in the presence of apamin but not TRAM-34. Western blot did not detect any differences in connexins 40 or 43 or in IKCa expression between male and female PCAs. CONCLUSIONS AND IMPLICATIONS: H2O2 mediated some part of endothelium-dependent vasorelaxation in female PCAs and EDH was more important in females, with differences in the contribution of gap junctions and IKCa channels. These findings may contribute to understanding vascular protection in premenopausal women

AGS and other tissue culture cells can unknowingly be persistently infected with PIV5; A virus that blocks interferon signalling by degrading STAT1

AbstractWhilst screening various cell lines for their ability to respond to interferon (IFN), we noted that in comparison to other tissue culture cells AGS tumour cells, which are widely used in biomedical research, had very low levels of STAT1. Subsequent analysis showed that the reason for this is that AGS cells are persistently infected with parainfluenza virus type 5 (PIV5; formally known as SV5), a virus that blocks the interferon (IFN) response by targeting STAT1 for proteasome-mediated degradation. Virus protein expression in AGS is altered in comparison to the normal pattern of virus protein synthesis observed in acutely infected cells, suggesting that the AGS virus is defective. We discuss the relevance of these results in terms of the need to screen cell lines for persistent virus infections that can alter cellular functions

Comparative analysis of classic brain component sizes in relation to flightiness in birds

Increased encephalization has been linked to a range of behavioural traits and scenarios. However, studies of whole brain size in this context have been criticised for ignoring the role of specific brain areas in controlling behaviour. In birds, the response to potential threats is one such behaviour that may relate to the way in which the brain processes sensory information. We used a phylogenetic generalised least squares (PGLS) analyses, based on five different phylogenetic hypotheses, to analyse the relationship of relative sizes of whole brain and brain components with Flight-Initiation Distance (FID), the distance at which birds flee from an approaching human, for 41 bird species. Starting distance (the distance at which an approach to a bird commences), body mass and eye size have elsewhere been shown to be positively associated with FID, and consequently were included as covariates in our analysis. Starting distance and body mass were by far the strongest predictors of FID. Of all brain components, cerebellum size had the strongest predictor weight and was negatively associated with FID but the confidence intervals on the average estimate included zero and the overall predictor weight was low. Models featuring individual brain components were generally more strongly weighted than models featuring whole brain size. The PGLS analyses estimated there to be no phylogenetic signal in the regression models, and hence produced results equivalent to ordinary least squares regression analysis. However analyses that assumed strong phylogenetic signal produced substantially different results with each phylogeny, and overall suggest a negative relationship between forebrain size and FID. Our analyses suggest that the evolutionary assumptions of the comparative analysis, and consideration of starting distance make a profound difference to the interpretation of the effect of brain components on FID in birds

ENGINEERING AN OLEOGINOUS YEAST FOR THE PRODUCTION OF BIODIESEL

poster abstractThere are economic and social interests in replacing the current energy dependence we have on petroleum-based oleochemicals. Yarrowia lipolytica, an oleaginous yeast, has the ability to metabolize unique carbon sources, particularly hydrocarbons and to accumulate large amounts of lipids which could be developed into a source of biodiesel. The ability of Y. lipolytica to accumulate triacylglycerols in lipid droplets and the complete sequencing of its genome make Y. lipolytica a viable organism to genetically engineer for the production of large quantities of biodiesel precursors. The purpose of this project is to genetically modify Y. lipolytica to further increase its production of triacylglycerols by knocking out genes that encode enzymes involved in the β-oxidation of fatty acids. This genetic modification will be accomplished by using homologous recombination to disrupt the genes POX3-5 and POT1. The 5’ and 3’ untranslated regions of POX3-5 and POT1 were amplified by polymerase chain reaction and cloned to allow a drug resistance gene to be introduced between them. Following cloning, these genes will be knocked out from the Y. lipolytica genome using drug resistance as a marker. The disruption of these genes is expected to increase the accumulation of triacylglycerols in Y. lipolytica lipid droplets versus the wild-type. Progress towards the goals of this project will be reported

Heterocellular induction of interferon by negative-sense RNA viruses

The infection of cells by RNA viruses is associated with the recognition of virus PAMPs (pathogen-associated molecular patterns) and the production of type I interferon (IFN). To counter this, most, if not all, RNA viruses encode antagonists of the IFN system. Here we present data on the dynamics of IFN production and response during developing infections by paramyxoviruses, influenza A virus and bunyamwera virus. We show that only a limited number of infected cells are responsible for the production of IFN, and that this heterocellular production is a feature of the infecting virus as opposed to an intrinsic property of the cells

Signatures of Relativistic Neutrinos in CMB Anisotropy and Matter Clustering

We present a detailed analytical study of ultra-relativistic neutrinos in cosmological perturbation theory and of the observable signatures of inhomogeneities in the cosmic neutrino background. We note that a modification of perturbation variables that removes all the time derivatives of scalar gravitational potentials from the dynamical equations simplifies their solution notably. The used perturbations of particle number per coordinate, not proper, volume are generally constant on superhorizon scales. In real space an analytical analysis can be extended beyond fluids to neutrinos. The faster cosmological expansion due to the neutrino background changes the acoustic and damping angular scales of the cosmic microwave background (CMB). But we find that equivalent changes can be produced by varying other standard parameters, including the primordial helium abundance. The low-l integrated Sachs-Wolfe effect is also not sensitive to neutrinos. However, the gravity of neutrino perturbations suppresses the CMB acoustic peaks for the multipoles with l>~200 while it enhances the amplitude of matter fluctuations on these scales. In addition, the perturbations of relativistic neutrinos generate a *unique phase shift* of the CMB acoustic oscillations that for adiabatic initial conditions cannot be caused by any other standard physics. The origin of the shift is traced to neutrino free-streaming velocity exceeding the sound speed of the photon-baryon plasma. We find that from a high resolution, low noise instrument such as CMBPOL the effective number of light neutrino species can be determined with an accuracy of sigma(N_nu) = 0.05 to 0.09, depending on the constraints on the helium abundance.Comment: 38 pages, 7 figures. Version accepted for publication in PR

Left-right symmetry in 5D and neutrino mass in TeV scale gravity models

We construct a left-right symmetric model based on the gauge group $SU(2)_L\times SU(2)_R\times U(1)_{B-L}$ in five dimensions where both the gauge bosons and fermions reside in all five dimensions. The orbifold boundary conditions are used not only to break the gauge symmetry down to $SU(2)_L\times U(1)_Y\times U(1)_{Y'}$ but also to ``project'' the right handed neutrino out of the zero mode part of the spectrum, providing a new way to understand the small neutrino masses without adding (singlet) bulk neutrinos. This formulation of the left-right model has also two new features: (i) it avoids most existing phenomenological bounds on the scale of the right handed $W_R$ boson allowing for the possibility that the right handed gauge bosons could have masses under a TeV, and (ii) it predicts a stable lepton with mass of order of the inverse radius of the fifth dimension.Comment: 20 pages; some new materials and references adde

Lepton Flavour Violating Leptonic/Semileptonic Decays of Charged Leptons in the Minimal Supersymmetric Standard Model

We consider the leptonic and semileptonic (SL) lepton flavour violating (LFV) decays of the charged leptons in the minimal supersymmetric standard model (MSSM). The formalism for evaluation of branching fractions for the SL LFV charged-lepton decays with one or two pseudoscalar mesons, or one vector meson in the final state, is given. Previous amplitudes for the SL LFV charged-lepton decays in MSSM are improved, for instance the $\gamma$-penguin amplitude is corrected to assure the gauge invariance. The decays are studied not only in the model-independent formulation of the theory in the frame of MSSM, but also within the frame of the minimal supersymmetric SO(10) model within which the parameters of the MSSM are determined. The latter model gives predictions for the neutrino-Dirac Yukawa coupling matrix, once free parameters in the model are appropriately fixed to accommodate the recent neutrino oscillation data. Using this unambiguous neutrino-Dirac Yukawa couplings, we calculate the LFV leptonic and SL decay processes assuming the minimal supergravity scenario. A very detailed numerical analysis is done to constrain the MSSM parameters. Numerical results for SL LFV processes are given, for instance for tau -> e (mu) pi0, tau -> e (mu) eta, tau -> e (mu) eta', tau -> e (mu) rho0, tau -> e (mu) phi, tau -> e (mu) omega, etc.Comment: 36 pages, 3 tables, 5 .eps figure

Late-time Entropy Production from Scalar Decay and Relic Neutrino Temperature

Entropy production from scalar decay in the era of low temperatures after neutrino decoupling will change the ratio of the relic neutrino temperature to the CMB temperature, and, hence, the value of N_eff, the effective number of neutrino species. Such scalar decay is relevant to reheating after thermal inflation, proposed to dilute massive particles, like the moduli and the gravitino, featuring in supersymmetric and string theories. The effect of such entropy production on the relic neutrino temperature ratio is calculated in a semi-analytic manner, and a recent lower bound on this ratio, obtained from the WMAP satellite and 2dF galaxy data, is used to set a lower bound of ~ 1.5 x 10^-23 Gev on the scalar decay constant, corresponding to a reheating temperature of about 3.3 Mev.Comment: 13 pages, to appear in PR