VP24-Karyopherin alpha binding affinities differ between Ebolavirus species, nfluencing interferon inhibition and VP24 stability

Zaire ebolavirus (EBOV), Bundibugyo ebolavirus (BDBV), and Reston ebolavirus (RESTV) belong to the same genus but exhibit different virulence properties. VP24 protein, a structural protein present in all family members, blocks interferon (IFN) signaling and likely contributes to virulence. Inhibition of IFN signaling by EBOV VP24 (eVP24) involves its interaction with the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear transporters. Here, we evaluated eVP24, BDBV VP24 (bVP24), and RESTV VP24 (rVP24) interactions with three NPI-1 subfamily KPNAs (KPNA1, KPNA5, and KPNA6). Using purified proteins, we demonstrated that each VP24 binds to each of the three NPI-1 KPNAs. bVP24, however, exhibited approximately 10-fold-lower KPNA binding affinity than either eVP24 or rVP24. Cell-based assays also indicate that bVP24 exhibits decreased KPNA interaction, decreased suppression of IFN induced gene expression, and a decreased half-life in transfected cells compared to eVP24 or rVP24. Amino acid sequence alignments between bVP24 and eVP24 also identified residues within and surrounding the previously defined eVP24-KPNA5 binding interface that decrease eVP24-KPNA affinity or bVP24-KPNA affinity. VP24 mutations that lead to reduced KPNA binding affinity also decrease IFN inhibition and shorten VP24 half-lives. These data identify novel functional differences in VP24-KPNA interaction and reveal a novel impact of the VP24-KPNA interaction on VP24 stability. IMPORTANCE The interaction of Ebola virus (EBOV) VP24 protein with host karyopherin alpha (KPNA) proteins blocks type I interferon (IFN) signaling, which is a central component of the host innate immune response to viral infection. Here, we quantitatively compared the interactions of VP24 proteins from EBOV and two members of the Ebolavirus genus, Bundibugyo virus (BDBV) and Reston virus (RESTV). The data reveal lower binding affinity of the BDBV VP24 (bVP24) for KPNAs and demonstrate that the interaction with KPNA modulates inhibition of IFN signaling and VP24 stability. The effect of KPNA interaction on VP24 stability is a novel functional consequence of this virus-host interaction, and the differences identified between viral species may contribute to differences in pathogenesis

Dysregulated NK cell PLCγ2 signaling and activity in juvenile dermatomyositis

Juvenile dermatomyositis (JDM) is a debilitating pediatric autoimmune disease manifesting with characteristic rash and muscle weakness. To delineate signaling abnormalities in JDM, mass cytometry was performed with PBMCs from treatment-naive JDM patients and controls. NK cell percentages were lower while frequencies of naive B cells and naive CD4+ T cells were higher in JDM patients than in controls. These cell frequency differences were attenuated with cessation of active disease. A large number of signaling differences were identified in treatment-naive JDM patients compared with controls. Classification models incorporating feature selection demonstrated that differences in phospholipase Cγ2 (PLCγ2) phosphorylation comprised 10 of 12 features (i.e., phosphoprotein in a specific immune cell subset) distinguishing the 2 groups. Because NK cells represented 5 of these 12 features, further studies focused on the PLCγ2 pathway in NK cells, which is responsible for stimulating calcium flux and cytotoxic granule movement. No differences were detected in upstream signaling or total PLCγ2 protein levels. Hypophosphorylation of PLCγ2 and downstream mitogen-activated protein kinase-activated protein kinase 2 were partially attenuated with cessation of active disease. PLCγ2 hypophosphorylation in treatment-naive JDM patients resulted in decreased calcium flux. The identification of dysregulation of PLCγ2 phosphorylation and decreased calcium flux in NK cells provides potential mechanistic insight into JDM pathogenesis

Paramagnetic Meissner effect in superconductors from self-consistent solutions of Ginzburg-Landau equations

The paramagnetic Meissner effect (PME) is observed in small superconducting samples, and a number of controversial explanations of this effect are proposed, but there is as yet no clear understanding of its nature. In the present paper PME is considered on the base of the Ginzburg-Landau theory (GL). The one-dimensional solutions are obtained in a model case of a long superconducting cylinder for different cylinder radii R, the GL-parameters \kappa and vorticities m. Acording to GL-theory, PME is caused by the presence of vortices inside the sample. The superconducting current flows around the vortex to screeen the vortex own field from the bulk of the sample. Another current flows at the boundary to screen the external field H from entering the sample. These screening currents flow in opposite directions and contribute with opposite signs to the total magnetic moment (or magnetization) of the sample. Depending on H, the total magnetization M may be either negative (diamagnetism), or positive (paramagnetism). A very complicated saw-like dependence M(H) (and other characteristics), which are obtained on the base of self-consistent solutions of the GL-equations, are discussed.Comment: 6 pages, 5 figures, RevTex, submitted to Phys. Rev.

Glycolytic requirement for NK cell cytotoxicity and cytomegalovirus control

NK cell activation has been shown to be metabolically regulated in vitro; however, the role of metabolism during in vivo NK cell responses to infection is unknown. We examined the role of glycolysis in NK cell function during murine cytomegalovirus (MCMV) infection and the ability of IL-15 to prime NK cells during CMV infection. The glucose metabolism inhibitor 2-deoxy-ᴅ-glucose (2DG) impaired both mouse and human NK cell cytotoxicity following priming in vitro. Similarly, MCMV-infected mice treated with 2DG had impaired clearance of NK-specific targets in vivo, which was associated with higher viral burden and susceptibility to infection on the C57BL/6 background. IL-15 priming is known to alter NK cell metabolism and metabolic requirements for activation. Treatment with the IL-15 superagonist ALT-803 rescued mice from otherwise lethal infection in an NK-dependent manner. Consistent with this, treatment of a patient with ALT-803 for recurrent CMV reactivation after hematopoietic cell transplant was associated with clearance of viremia. These studies demonstrate that NK cell-mediated control of viral infection requires glucose metabolism and that IL-15 treatment in vivo can reduce this requirement and may be effective as an antiviral therapy

Lack of complex I is associated with oncocytic thyroid tumours

Oncocytic tumours are characterised by hyperproliferation of mitochondria. We immunohistochemically analysed all enzymes of the oxidative phosphorylation system in 19 oncocytic thyroid tumours. A specific lack of complex I was detected, which was expressed at <5% of the level determined in surrounding non-cancerous tissue

Anisotropy in mechanical properties and fracture behavior of an oxide dispersion Fe20Cr5Al alloy

Anisotropy of fracture toughness and fracture behavior of Fe20Cr5Al oxide dispersion-strengthened alloy has been investigated by means of compression tests, hardness tests, and wedge splitting test. The results show a small effect of the compression direction on yield strength (YS) and strain hardening. The YS is minimum for longitudinal direction and maximum for the tangential direction. The transverse plastic strain ratio is similar for tangential and longitudinal directions but very different from that in normal direction. Hardness depends on the indentation plane; it is lower for any plane parallel to the L-T plane and of similar magnitude for the other orthogonal planes, i.e., the L-S and T-S planes. Macroscopically, two failure modes have been observed after wedge-splitting tests, those of LS and TS specimens in which fracture deviates along one or two branches normal to the notch plane, and those of LT, TL, SL, and ST specimens in which fracture propagates along the notch plane. Besides LT and TL specimens present delaminations parallel to L-T plane. Both, the fracture surface of branching cracks and that of the delaminations, show an intergranular brittle fracture appearance. It is proposed that the main cause of the delamination and crack branching is the alignment in the mesoscopic scale of the ultrafine grains structure which is enhanced by the 〈110〉- texture of the material and by the presence in the grain boundaries of both yttria dispersoids and impurity contaminations. An elastoplastic finite element analysis was performed to study what stress state is the cause of the branches and delaminations. It is concluded that the normal to the crack branches and/or the shear stress components could determine the crack bifurcation mechanism, whereas the delamination it seems that it is controlled by the magnitude of the stress component normal to the delamination plane. © The Minerals, Metals & Materials Society and ASM International 2014.Peer Reviewe