Characterization of polyoxometalate I as an inhibitor of RNA-dependent RNA polymerase of Foot and Mouth Disease virus [abstract]

Abstract only availableFoot and Mouth Disease (FMD) is a highly contagious disease that affects a variety of domesticated cloven-hoofed animals including cattle, swine, sheep and goats, as well as several wild animal species. FMD outbreaks are currently controlled with mass-extermination of livestock. The financial cost of potential outbreaks would be immense. This disease is caused by foot-and-mouth disease virus (FMDV), a non-enveloped, single-stranded, positive-sense RNA virus. The purpose of our investigation is to identify chemicals that interfere with the replication of FMDV. As part of this effort we have identified a polyoxometalate inhibitor (polyoxometalate I). We have cloned, expressed and purified FMDV RdRp. We use steady-state kinetic experiments and polymerization assays to characterize the inhibitory activity of the polyoxometalate I, determining the precise inhibitory potential and the mechanism of inhibition. Preliminary results show that polyoxometalate I inhibits the FMDV RdRp surprisingly efficiently with an IC50 of 0.5uM. Current experiments are focusing on a detailed kinetic characterization of the mechanism of action for this inhibitor. This research may provide insights that lead to new treatment options to prevent the further spread of FMD to unaffected animals.USD

Subsurface Pore Water Contributions to Stream Concentration-Discharge Relations Across a Snowmelt Hydrograph

This study investigated the concentration discharge (C-Q) patterns of selected elements transported to streams during spring snowmelt through an instrumented mixed-conifer forested catchment in rhyolitic terrain in the Jemez Mountains (NM, United States). High frequency, concurrent sampling of soil solution and gasses, groundwater, and surface water enabled identification and sourcing of five groups of solutes with distinct C-Q behavior. Non-hydrolyzing cations and strong acid anions, had mostly positive C-Q relations and a clockwise hysteresis pattern related to flushing of a limited reservoir of solutes accumulated in soils under snowpack. Rare earth elements (REEs) and dissolved organic carbon (DOC), demonstrated large positive C-Q relations and a clockwise hysteresis pattern, consistent with their co-transport as metal-ligand complexes, and signaling biologically induced weathering reactions in the soil. Silicon and dissolved inorganic carbon (DIC) exhibited chemostatic C-Q trends and an anti-clockwise hysteresis pattern consistent with sourcing from deep groundwater. Hydrolyzing metals (Mn, Al, Ti, and Zr) with high coefficients of variance (CV) for concentration relative to CV values for stream discharge and with no significant C-Q pattern, were found to be transported mainly as filterable colloids. Fe C-Q behavior was similar to this hydrolyzing metals group, but complexation with DOC was also important for Fe during the initial stage of the snowmelt hydrograph. Investigation of time-series of solutes and gasses provided evidence for biologically induced silicate weathering reactions that initiated in the soil subsurface and propagated down through groundwater to streams

Evaluating clinician acceptability of the prototype CanRisk tool for predicting risk of breast and ovarian cancer: A multi-methods study.

BACKGROUND:There is a growing focus on the development of multi-factorial cancer risk prediction algorithms alongside tools that operationalise them for clinical use. BOADICEA is a breast and ovarian cancer risk prediction model incorporating genetic and other risk factors. A new user-friendly Web-based tool (CanRisk.org) has been developed to apply BOADICEA. This study aimed to explore the acceptability of the prototype CanRisk tool among two healthcare professional groups to inform further development, evaluation and implementation. METHOD:A multi-methods approach was used. Clinicians from primary care and specialist genetics clinics in England, France and Germany were invited to use the CanRisk prototype with two test cases (either face-to-face with a simulated patient or via a written vignette). Their views about the tool were examined via a semi-structured interview or equivalent open-ended questionnaire. Qualitative data were subjected to thematic analysis and organised around Sekhon's Theoretical Framework of Acceptability. RESULTS:Seventy-five clinicians participated, 21 from primary care and 54 from specialist genetics clinics. Participants were from England (n = 37), France (n = 23) and Germany (n = 15). The prototype CanRisk tool was generally acceptable to most participants due to its intuitive design. Primary care clinicians were concerned about the amount of time needed to complete, interpret and communicate risk information. Clinicians from both settings were apprehensive about the impact of the CanRisk tool on their consultations and lack of opportunities to interpret risk scores before sharing them with their patients. CONCLUSIONS:The findings highlight the challenges associated with developing a complex tool for use in different clinical settings; they also helped refine the tool. This prototype may not have been versatile enough for clinical use in both primary care and specialist genetics clinics where the needs of clinicians are different, emphasising the importance of understanding the clinical context when developing cancer risk assessment tools

Inhibitors of Foot and Mouth Disease Virus Targeting a Novel Pocket of the RNA-Dependent RNA Polymerase

Foot-and-Mouth Disease Virus (FMDV) is a picornavirus that infects cloven-hoofed animals and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination requires at least 7 days to trigger an effective immune response. There are currently no approved inhibitors for the treatment or prevention of FMDV infections.Using a luciferase-based assay we screened a library of compounds and identified seven novel inhibitors of 3Dpol, the RNA-dependent RNA polymerase of FMDV. The compounds inhibited specifically 3Dpol (IC(50)s from 2-17 µM) and not other viral or bacterial polymerases. Enzyme kinetic studies on the inhibition mechanism by compounds 5D9 and 7F8 showed that they are non-competitive inhibitors with respect to NTP and nucleic acid substrates. Molecular modeling and docking studies into the 3Dpol structure revealed an inhibitor binding pocket proximal to, but distinct from the 3Dpol catalytic site. Residues surrounding this pocket are conserved among all 60 FMDV subtypes. Site directed mutagenesis of two residues located at either side of the pocket caused distinct resistance to the compounds, demonstrating that they indeed bind at this site. Several compounds inhibited viral replication with 5D9 suppressing virus production in FMDV-infected cells with EC(50) = 12 µM and EC(90) = 20 µM).We identified several non-competitive inhibitors of FMDV 3Dpol that target a novel binding pocket, which can be used for future structure-based drug design studies. Such studies can lead to the discovery of even more potent antivirals that could provide alternative or supplementary options to contain future outbreaks of FMD

Bridging Alone: Religious Conservatism, Marital Homogamy, and Voluntary Association Membership

This study characterizes social insularity of religiously conservative American married couples by examining patterns of voluntary associationmembership. Constructing a dataset of 3938 marital dyads from the second wave of the National Survey of Families and Households, the author investigates whether conservative religious homogamy encourages membership in religious voluntary groups and discourages membership in secular voluntary groups. Results indicate that couples’ shared affiliation with conservative denominations, paired with beliefs in biblical authority and inerrancy, increases the likelihood of religious group membership for husbands and wives and reduces the likelihood of secular group membership for wives, but not for husbands. The social insularity of conservative religious groups appears to be reinforced by homogamy—particularly by wives who share faith with husbands

Novel inhibitors of Foot and Mouth Disease Virus (FMDV) Targeting the RNA-Dependent RNA Polymerase activity of 3Dpol [abstract]

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionFoot-and-Mouth Disease Virus (FMDV) is a positive stranded picornavirus which infects cloven-hoofed animals, such as cattle, pigs and sheep, and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination could be used but it would require at least 7 days to trigger an effective immune response. On the contrary, the use of antiviral drugs is expected to have prophylactic and/or therapeutic effects almost immediately. However, there are currently no approved FMDV inhibitors. Here we have applied a combination of screening, biochemical, virological, and molecular modeling tools to discover, validate, and characterize novel inhibitors of FMDV replication. Using a luciferase-based assay we have screened a chemical library of compounds and have identified two compounds, 5-chloro-3-(thiophen-2-yl-sulfanylmethyl)-1-benzothiophene 1,1-dioxide (or C7F8) and N'1-thieno[2,3-d]pyrimidin-4-yl-4-chloro-1-benzenesulfonohydrazide (or C5D9) that inhibited the RNA-dependent RNA polymerase activity of FMDV replicase (3Dpol) with IC50 values of 2.5 μM and 15 μM respectively. These compounds were shown to be specific inhibitors of FMDV 3Dpol and not nucleic acid chelators, as they did not affect activity of other viral polymerases using the same nucleic acid substrate. Molecular modeling docking experiments suggest that both inhibitors bind at a pocket proximal to, but distinct from, the NTP binding site of 3Dpol, thereby affecting indirectly RNA synthesis. C7F8 and C5d9 were not cytotoxic at concentrations up to at least 100 ||M. Importantly, C5D9 exhibited antiviral activity and suppressed virus production in FMDV-infected cells with 50% and 90% effective concentrations (EC50 and EC90) of 10 ||M and 20 ||M, respectively. The results indicate that 3Dpol inhibitors can be promising anti-FMDV agents for use as alternative or supplementary options to contain future outbreaks of FMD

Use of cancer-specific yeast-secreted in vivo biotinylated recombinant antibodies for serum biomarker discovery

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

Atypical Social Modulation of Imitation in Autism Spectrum Conditions.

Appropriate modulation of imitation according to social context is important for successful social interaction. In the present study we subliminally primed high-functioning adults with ASC and age- and IQ-matched controls with either a pro- or non- social attitude. Following priming, an automatic imitation paradigm was used to acquire an index of imitation. Whereas imitation levels were higher for pro-socially primed relative to non-socially primed control participants, there was no difference between pro- and non- socially primed individuals with ASC. We conclude that high-functioning adults with ASC demonstrate atypical social modulation of imitation. Given the importance of imitation in social interaction we speculate that difficulties with the modulation of imitation may contribute to the social problems characteristic of ASC