The SARS-CoV-2 pandemic has made it clear that we have a desperate need for antivirals. We present work that the mammalian SKI complex is a broad-spectrum, host-directed, antiviral drug target. Yeast ...suppressor screening was utilized to find a functional genetic interaction between proteins from influenza A virus (IAV) and Middle East respiratory syndrome coronavirus (MERS-CoV) with eukaryotic proteins that may be potential host factors involved in replication. This screening identified the SKI complex as a potential host factor for both viruses. In mammalian systems siRNA-mediated knockdown of SKI genes inhibited replication of IAV and MERS-CoV. In silico modeling and database screening identified a binding pocket on the SKI complex and compounds predicted to bind. Experimental assays of those compounds identified three chemical structures that were antiviral against IAV and MERS-CoV along with the filoviruses Ebola and Marburg and two further coronaviruses, SARS-CoV and SARS-CoV-2. The mechanism of antiviral activity is through inhibition of viral RNA production. This work defines the mammalian SKI complex as a broad-spectrum antiviral drug target and identifies lead compounds for further development.
Interferon inducible transmembrane proteins (IFITMs) are broad‐spectrum antiviral factors. In cell culture the entry of many enveloped viruses, including orthomyxo‐, flavi‐, and filoviruses, is ...inhibited by IFITMs, though the mechanism(s) involved remain unclear and may vary between viruses. We demonstrate that Sindbis and Semliki Forest virus (SFV), which both use endocytosis and acid‐induced membrane fusion in early endosomes to infect cells, are restricted by the early endosomal IFITM3. The late endosomal IFITM2 is less restrictive and the plasma membrane IFITM1 does not inhibit normal infection by either virus. IFITM3 inhibits release of the SFV capsid into the cytosol, without inhibiting binding, internalization, trafficking to endosomes or low pH‐induced conformational changes in the envelope glycoprotein. Infection by SFV fusion at the cell surface was inhibited by IFITM1, but was equally inhibited by IFITM3. Furthermore, an IFITM3 mutant (Y20A) that is localized to the plasma membrane inhibited infection by cell surface fusion more potently than IFITM1. Together, these results indicate that IFITMs, in particular IFITM3, can restrict alphavirus infection by inhibiting viral fusion with cellular membranes. That IFITM3 can restrict SFV infection by fusion at the cell surface equivalently to IFITM1 suggests that IFITM3 has greater antiviral potency against SFV.
IFITM3 can inhibit cellular infection by two alphaviruses, Sindbis and Semliki Forest virus (SFV). IFITM2 has lower antiviral activity, and IFITM1 does not restrict either virus through the endocytic route of infection. IFITM3 does not inhibit SFV binding, internalization into cells, trafficking to endosomes or low pH‐induced conformational changes in the E1 fusion protein. However, the viral capsid is not released into the cytosol in IFITM3 expressing cells.
Southern Hemisphere Maser Astrometry Ellingsen, Simon; Reid, Mark; Menten, Karl ...
Proceedings of the International Astronomical Union,
12/2022, Letnik:
18, Številka:
S380
Journal Article
Recenzirano
Odprti dostop
Many astrophysical phenomena can only be studied in detail for objects in our galaxy, the Milly Way, but we know much more about the structure of thousands of nearby galaxies than we do about our own ...Galaxy. Accurate distance measurements in the Milky Way underpin our ability to understand a wide range of astrophysical phenomena and this requires observations from both the northern and southern hemisphere. Our ability to measure accurate parallaxes to southern masers has been hampered a range of factors, in particular the absence of a dedicated, homogeneous VLBI array in the south. We have recently made significant advances in astrometric calibration techniques which allow us to achieve trigonometric parallax accuracies of around 10 micro-arcseconds (μas) for 6.7 GHz methanol masers with a hetrogeneous array of 4 antennas. We outline the details of this new “multiview” technique and present the first trigonometric parallax measurements that utilise this approach.
Starting from a yeast suppressor screening platform, we have identified the SKI complex as a potential broad-spectrum antiviral target. We found that the NS1 protein of influenza A virus (IAV) and ...the ORF4a protein of Middle East respiratory syndrome coronavirus (MERS-CoV), which both function to bind double-strand RNA and inhibit cellular interferon responses, cause a slow growth phenotype when expressed in yeast. Knockout of the components of the yeast SKI complex caused a loss of this slow growth phenotype, suggesting a functional link between the viral proteins and the SKI complex. The SKI complex is a helicase that unwinds double-strand RNA and sends it to the RNA exosome for degradation. We next investigated whether the highly conserved human SKI complex was important for replication of IAV and MERS-CoV. RNAi based experiments showed that both viruses were inhibited when the SKI complex was removed, suggesting the complex has a proviral role in replication. Through in silico modelling using the published crystal structure of the SKI complex, we looked for potential binding pockets for chemical compounds. We screened a selection of these compounds for antiviral activity and have found four different chemicals capable of inhibiting IAV infection. Our most studied of these also inhibits not only MERS-CoV, but also Ebolavirus Makona. Our data suggests the SKI complex may be a target for broad-spectrum antiviral therapy and we have multiple chemical structures from which to work to develop therapeutic approaches.
Hemorrhage leads to 30% to 40% of trauma deaths, with up to 50% of these deaths transpiring before hospital arrival. There is a growing amount of experience and research with prehospital blood ...administration, but few tools exist to identify the need and impact of a prehospital blood program in a community. Validating a blood use prediction tool locally will allow us to apply that validation to prehospital patients in other communities. Multiple algorithmic scoring tools that predicted the use of blood products were assessed using data from Baylor Scott and White Memorial Hospital, a level I trauma center. A total of 100 men and 51 women were included in the study, 99 of whom received a blood transfusion within 2 hours of hospital arrival. Comparing the scoring systems using our internal data, we found that three scoring systems were approximately equal at determining the need for blood products: Criteria A for the Zhu et al scoring system had a specificity and positive predictive value (PPV) of 92% while maintaining a sensitivity and negative predictive value (NPV) of 48%. Similarly, the EBTNS scoring system with a cutoff of ≥6 resulted in a specificity of 90%, PPV of 91%, sensitivity of 56%, and NPV of 52%. Lastly, the ABC scoring system with a cutoff of ≥2 had a specificity of 94%, PPV of 91%, sensitivity of 38%, and NPV of 56%. These scoring tools can be used in the prehospital setting to predict the need for blood in geographic areas in order to help with asset utilization.
Host interferon-induced transmembrane proteins (IFITMs) are broad-spectrum antiviral restriction factors. Of these, IFITM3 potently inhibits viruses that enter cells through acidic endosomes, many of ...which are zoonotic and emerging viruses with bats (order Chiroptera) as their natural hosts. We previously demonstrated that microbat IFITM3 is antiviral. Here, we show that bat IFITMs are characterized by strong adaptive evolution and identify a highly variable and functionally important site-codon 70-within the conserved CD225 domain of IFITMs. Mutation of this residue in microbat IFITM3 impairs restriction of representatives of four different virus families that enter cells via endosomes. This mutant shows altered subcellular localization and reduced S-palmitoylation, a phenotype copied by mutation of conserved cysteine residues in microbat IFITM3. Furthermore, we show that microbat IFITM3 is S-palmitoylated on cysteine residues C71, C72, and C105, mutation of each cysteine individually impairs virus restriction, and a triple C71A-C72A-C105A mutant loses all restriction activity, concomitant with subcellular re-localization of microbat IFITM3 to Golgi-associated sites. Thus, we propose that S-palmitoylation is critical for Chiropteran IFITM3 function and identify a key molecular determinant of IFITM3 S-palmitoylation.
Core supported study of the heterogeneous Ungani dolomite reservoir architecture is driving development drilling and upgrades to field resource estimates. Vuggy connected and macro non-connected pore ...space was directly measured over a 70m continuous core using 3D structural analysis of CT-scans. Plug density measurements indicate non-connected or sub-140 micron resolution contribution of around 1% to 2.5% (pu) for the tight matrix, but all remaining porosity potentially contributes to oil production. The high resolution core porosity data is vertically repositioned and upscaled to calibrate neutron-density and sonic petrophysically derived porosities which are inadequate to resolve productive zones using conventional reservoir cut-offs. Conditioned resistivity image data correlated exceptionally with directly measured connected porosities. Reservoir properties were extrapolated to all wells across the Ungani field giving field net/gross estimates of up to 63% and porosities over 30% pu in some vuggy and brecciaed zones. The heterogeneity and prolific nature of the uppermost 17m of reservoir had not been previously recognised due to poor log data coverage and access at the casing points. Recent re-analysis of this section at Ungani-3 with Chemostrat ICP-OES-MS analysis of ditch cuttings was instrumental in proposing additional drilling to re-target this zone. Mineralogy analysis is used to calculate rock grain densities and help calibrate neutron-density derived porosity logs over the Ungani field. Up-scaled core porosity correlates well with density and sonic porosity logs. Resistivity logs adjusted for minerology can also be used to predict porosity and support the use of resistivity image logs to identify vuggy zones and estimate porosity at a higher resolution than conventional logging tools. A field static model was populated with three facies distributed over vertical zones according to the distribution encountered in the core porosity analysis and well logs, and iteratively matched to the dynamic pressure data and field production history which exhibits field scale multi-Darcy horizontal permeability and protection from vertical water cut. Further drilling and downhole artificial lift is planned to extend field production rates to 3000 bbls/day. Increased confidence in this regionally developed reservoir is supporting further exploration of undrilled prospects in this immature and under-explored trend
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