One challenge for multisite clinical trials is ensuring that the conditions of an informative trial are incorporated into all aspects of trial planning and execution. The multicenter model can provide the potential for a more informative environment, but it can also place a trial at risk of becoming uninformative due to lack of rigor, quality control, or effective recruitment, resulting in premature discontinuation and/or non-publication. Key factors that support informativeness are having the right team and resources during study planning and implementation and adequate funding to support performance activities. This communication draws on the experience of the National Center for Advancing Translational Science (NCATS) Trial Innovation Network (TIN) to develop approaches for enhancing the informativeness of clinical trials. We distilled this information into three principles: (1) assemble a diverse team, (2) leverage existing processes and systems, and (3) carefully consider budgets and contracts. The TIN, comprised of NCATS, three Trial Innovation Centers, a Recruitment Innovation Center, and 60+ CTSA Program hubs, provides resources to investigators who are proposing multicenter collaborations. In addition to sharing principles that support the informativeness of clinical trials, we highlight TIN-developed resources relevant for multicenter trial initiation and conduct.

OBJECTIVES/GOALS: Current COVID-19 rapid molecular tests require cartridge-reader detection, expensive circuitry, and complex microfluidics making the most accurate tests unavailable to the masses. Here we present a rapid molecular diagnostic leveraging isothermal amplification and paper-based microfluidics for a low-cost ultra-sensitive COVID-19 assay. METHODS/STUDY POPULATION: We designed a reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of SARS-CoV-2 and bacteriophage MS2 RNA. RT-RPA is a sequence specific, ultrasensitive, rapid isothermal DNA amplification technique that is well suited to home based testing due to its rapid assay time, robustness, ease of use, and readout options. RT-RPA reagents are added to a tube and incubated at 39°C in a fluorometer. Realtime fluorometer data gives results in under 15 minutes. This assay also provides visual detection via lateral flow readout with results in 23 minutes. RESULTS/ANTICIPATED RESULTS: We have developed a rapid multiplexed nucleic acid amplification assay with an internal process control for SARS-CoV-2 using single-pot RT-RPA. We screened 21 primer combinations to select primers that demonstrated excellent performance and target specificity against common respiratory viruses. We demonstrate the ability to multiplex SARS-CoV-2 and MS2 detection, utilizing MS2 as an internal process control for lysis, reverse transcription, amplification, and readout. We show duplexed detection using both fluorescence readout and visual readout using lateral flow strips. Duplexed fluorescence detection shows a limit of detection of 25 copies per reaction. Duplexed lateral flow readout shows a limit of detection of 50 copies per reaction DISCUSSION/SIGNIFICANCE: We developed a duplexed RT-RPA assay for SARS-CoV-2 with fluorescence or lateral flow readout. Our assay does not re-quire expensive reader, circuity, or fluid handling. The low material cost, temperature, and robustness make it ideal for a more accurate home-based COVID-19 diagnostic.

The objective of this study was to assess the effect of providing mats in the farrowing pen on behaviour and lesions in sows (n = 38) and piglets. Nineteen of the sows and their litters housed in farrowing crates with metal slatted floors were provided with mats, the remaining 19 acting as controls. Sow skin was inspected pre-farrowing and at weaning: the skin and feet of piglets were examined when they were 6 days old. Skin lesions were scored (on a scale of 0 to 6) according to severity. Total lesion scores per sow and median total lesion scores per litter were calculated. Sow behaviour was recorded by direct observation at feeding. Usage of a heat pad for lying in the presence or absence of mats was assessed over a 2hr observation period. Median skin lesion scores of sows did not differ between treatments. Litters from the control treatment had lower skin lesion scores (median [range] 8 [2-12] vs 10 [5-21]; P < 0.05). Median lesion scores of the front legs tended to be higher for piglets on mats (median [range] 8 [1.5-13] vs 4 [1.5-8]; ns). Sows on metal flooring went from kneeling to lowering their hindquarters faster (median [range] 15 [2—45] s) than sows on mats (23 [11—54] s) (P < 0.05). Furthermore, they slipped more on their front (median [range] 2.7 [0-8.7] vs 0 [0-1.3] slips min1; P < 0.001) and hind (median [range] 5.3 [0.3-31.7] vs 1.9 [0-33.3] slips min1; P < 0.05) feet while standing, than sows on mats. More piglets lay on the heat pad when it was covered by a mat (mean ± SEM 45.97 ± 4.49 %) than when no mat was provided (29.29 ± 4.47 %)(P < 0.05). Mats improve sow comfort and reduce slipping. They encourage piglets to lie on the heat pad but cause damage to the skin of the front legs. Providing pigs with mats in the farrowing house could greatly improve welfare on slatted floors, but further research is needed to identify materials that are less abrasive.

Leafy green vegetables are a common source of Shiga toxin-producing Escherichia coli O157:H7 (STEC O157) foodborne illness outbreaks. Ruminant animals, primarily cattle, are the major reservoir of STEC O157. Epidemiological, traceback and field investigations were conducted to identify potential outbreak sources. Product and environmental samples were tested for STEC. A reoccurring strain of STEC O157 caused two multistate outbreaks linked to romaine lettuce in 2018 and 2019, resulting in 234 illnesses in 33 states. Over 80% of patients interviewed consumed romaine lettuce before illness. The romaine lettuce was sourced from two California growing regions: Santa Maria and Salinas Valley in 2018 and Salinas Valley in 2019. The outbreak strain was isolated from environmental samples collected at sites >90 miles apart across growing regions, as well as from romaine-containing products in 2019. Although the definitive route of romaine contamination was undetermined, use of a contaminated agricultural water reservoir in 2018 and contamination from cattle grazing on adjacent land in 2019 were suspected as possible factors. Preventing lettuce contamination from growth to consumption is imperative to preventing illness. These outbreaks highlight the need to further understand mechanisms of romaine contamination, including the role of environmental or animal reservoirs for STEC O157.

Praziquantel (PZQ) is the drug of choice for schistosomiasis. The potential drug resistance necessitates the search for adjunct or alternative therapies to PZQ. Previous functional genomics has shown that RNAi inhibition of Ca2+/calmodulin-dependent protein kinase II (CaMKII) gene in Schistosoma adult worms significantly improved the effectiveness of PZQ. Here we tested the in vitro efficacy of 15 selective and non-selective CaMK inhibitors against Schistosoma mansoni and showed that PZQ efficacy was improved against refractory juvenile parasites when combined with these CaMK inhibitors. By measuring CaMK activity and the mobility of adult S. mansoni, we identified two non-selective CaMK inhibitors, Staurosporine (STSP) and 1Naphthyl PP1 (1NAPP1), as promising candidates for further study. The impact of STSP and 1NAPP1 was investigated in mice infected with S. mansoni in the presence or absence of a sub-lethal dose of PZQ against 2- and 7-day-old schistosomula and adults. Treatment with STSP/PZQ induced a significant (47–68%) liver egg burden reduction compared with mice treated with PZQ alone. The findings indicate that the combination of STSP and PZQ dosages significantly improved anti-schistosomal activity compared to PZQ alone, demonstrating the potential of selective and non-selective CaMK/kinase inhibitors as a combination therapy with PZQ in treating schistosomiasis.

Toxoplasma gondii rhoptry protein TgROP18 is a polymorphic virulence effector that targets immunity-related GTPases (IRGs) in rodents. Given that IRGs are uniquely diversified in rodents and not in other T. gondii intermediate hosts, the role of TgROP18 in manipulating non-rodent cells is unclear. Here we show that in human cells TgROP18I interacts with the interferon-gamma-inducible protein N-myc and STAT interactor (NMI) and that this is a property that is unique to the type I TgROP18 allele. Specifically, when expressed ectopically in mammalian cells only TgROP18I co-immunoprecipitates with NMI in IFN-γ-treated cells, while TgROP18II does not. In parasites expressing TgROP18I or TgROP18II, NMI only co-immunoprecipitates with TgROP18I and this is associated with allele-specific immunolocalization of NMI on the parasitophorous vacuolar membrane (PVM). We also found that TgROP18I reduces NMI association with IFN-γ-activated sequences (GAS) in the IRF1 gene promoter. Finally, we determined that polymorphisms in the C-terminal kinase domain of TgROP18I are required for allele-specific effects on NMI. Together, these data further define new host pathway targeted by TgROP18I and provide the first function driven by allelic differences in the highly polymorphic ROP18 locus.

The medium-band Vilnius photometric system with the mean wavelengths at 345 (U), 374 (P), 405 (X), 466 (Y), 516 (Z), 544 (V), and 656 (S) nm for many years was an important tool to determine interstellar reddenings and distances of single stars due to its ability to classify stars of all temperatures in spectral classes and luminosity classes in the presence of different interstellar reddenings. At present, Gaia DR2 presents distances to stars with an unprecedented accuracy at least up to 3 kpc. However, multicolor photometry, which allows the classification of stars as well as the preliminary determination of stellar temperatures, gravities, metallicities and interstellar reddenings, remains an important method for distant stars. Here we present an empirical calibration of the intrinsic color indices of the Vilnius system in terms of physical parameters of stars for dwarf and giant stars of spectral classes F-G-K-M. In any attempted photometric determination of physical parameters of stars it is important to have an extensive and homogeneous sample of spectroscopically determined parameters for stars for which there are also accurate photometric data. As a source catalogue for the Vilnius photometry the latest updated version of the Catalogue of Photoelectric Observations in the Vilnius System was used, which contains compilations from the published photometry for about 11 000 stars. The stars which had both the Gaia DR2 parallaxes and the determinations of stellar parameters from high-dispersion spectra were extracted from this catalogue. The final sample contains more than 1500 stars of spectral classes F-M. The majority of these stars (ca 70%) are not reddened, for others the values of interstellar reddening AV were determined using the regular techniques of photometric classification in the Vilnius system. The absolute magnitudes MV and consequently the luminosity classes were determined using Gaia DR2 parallaxes. We present the analytical expressions for the effective temperature Teff and surface gravity logg and evaluate the errors of solutions for dwarf and giant stars. To test the accuracy of the proposed method, we have compared our results with the stars observed by Gaia and with the stellar parameters available from the large spectroscopic surveys: APOGEE, Gaia-ESO, GALAH, LAMOST, RAVE and SEGUE. The results of comparison contain 5-6 % outliers.

The proposed method allows the fast and straightforward evaluation of stellar physical parameters for the stars observed in the Vilnius photometric system. Despite the fact, that the accuracy of determination is significantly lower than in the case of spectroscopic methods, the method described may be useful for distant faint stars, which are still inaccessible for spectroscopic observations.

A suite of experimentally deformed single-crystal pyrite samples has been investigated using electron backscatter diffraction (EBSD). Single crystals were loaded parallel to <100> or <110> and deformed at a strain rate of 10-5 s-1, confining pressure of 300 MPa and temperatures of 600°C and 700°C. Although geometrically (Schmid factor) the {001}<100> slip system should not be activated in <100> loaded samples, lattice rotation and boundary trace analyses of the distorted crystals indicate this slip system is easier to justify. Determination of 75 MPa as the critical resolved shear stress (CRSS) for {001}<100> activation, in the <110> loaded crystals, suggests a crystal misalignment of ~5—15° in the <100> loaded crystals would be sufficient to activate the {001}<100> slip system. Therefore, {001}<100> is considered the dominant slip system in all of the single-crystal pyrite samples studied. Slip-system analysis of the experimentally deformed polycrystalline pyrite aggregates is consistent with the single-crystal findings, with the exception that {001}<11̄> also appears to be important, although less common than the {001}<100> slip system. The lack of crystal preferred orientation (CPO) development in the polycrystalline pyrite aggregates can be accounted for by the presence of two independent symmetrically equivalent slip systems more than satisfying the von Mises criterion.

Understanding the deformation mechanisms that may operate in pyrite (FeS2) across a range of P-Tconditions is important in deciphering the history of deformed ore deposits. Pyrite has frequently been considered a hard mineral, which deforms by cataclastic flow or diffusive processes, if at all, at temperatures <425°C. However, utilizing SEM-based orientation-contrast (OC) imaging and electron-backscatter diffraction (EBSD) techniques, plastic deformation can now be readily identified within pyrite grains. In this study, a series of pyrite-rich polymetallic ore deposits, deformed at low temperature metamorphic conditions (∼200—420°C), have been investigated. Results indicate that pyrite grains in all the ore deposits preserve internal lattice ‘distortion’ or ‘bending’ and therefore plastic deformation mechanisms have operated. Many pyrite grains in the ore deposits also contain low-angle (∼2°) sub-grain boundaries or ‘dislocation walls', indicating that both dislocation glide and creep have been the dominant deformation mechanisms at peak metamorphic conditions within the pyrite grains. These results suggest that the brittle-ductile transition in pyrite occurs at temperatures potentially as low as ∼200°C, far lower than implied from previous studies or the current pyrite deformation-mechanism map.

The copper-bearing stratabound pyritic massive sulphide bodies contained in metamorphosed basic eruptives of Ordovician age at Sulitjelma in Nordland County, Norway, form one of the important fields of sulphide mineralisation within the Köli Nappe Complex. The sulphide bodies and their enclosing rocks were subject to successive stages of penetrative deformation and recrystallisation during the cycle of metamorphism and tectonic transport caused by the Scandian Orogeny. Textures within the ores and the immediate envelope of schists show that strain was focused along the mineralised horizons. The marked contrast in competence between the massive pyritic sulphides and their envelopes of alteration composed dominantly of phyllosilicates, and the metasediments of the overlying Furulund Group, led to the formation of macroscale fold and shear structures. On the mesoto microscale, a variety of textures have been formed within the pyrite-pyrrhotite-chalcopyrite-sphalerite sulphide rocks as a result of strain and recrystallisation. Variations in pyrite:pyrrhotite ratios and in the texture and proportions of associated gangue minerals evidently governed the strength and ductility of the sulphide rocks so that the same sulphide mineral can behave differently, displaying different textures in different matrices. In massive pyritic samples there is evidence of evolution towards textural equilibrium by recrystallisation, grain growth and annealment during the prograde part of the metamorphic cycle. Later, brittle deformation was superimposed on these early fabrics and the textural evidence is clearly preserved. By comparing published data on the brittle-ductile transformation boundaries of sulphide minerals with the conditions governing metamorphism at Sulitjelma, it is concluded that most of the brittle deformation in the sulphides took place during or after D3 under retrograde greenschist conditions. Grain growth of pyrite in matrices of more ductile sulphides during the prograde and early retrograde stages of metamorphism produced the coarse metablastic textures for which Sulitjelma is well-known. In some zones of high resolved shear stress, pyrite shows ductile behaviour which could be explained by a dislocation flow mechanism operating at conditions close to the metamorphic peak. In those horizons in which pyrrhotite is the dominant iron sulphide, the contrast in ductility between silicates, pyrite and pyrrhotite has led to the development of spectacular tectonoclastic textures in which fragments of wall rock have been broken, deformed, rolled and rotated within the ductile pyrrhotite matrix.

The shaping of PNe as a result of an interaction with a planet is a hypothesis that has been suggested for nearly two decades. However, exploring the idea observationally is challenging due to the lack of capabilities needed to detect any evidence of such a scenario. Nonetheless, we propose that the hypothesis can be indirectly tested via a combination of exoplanet formation and evolution theories, the star and planet formation histories of the galaxy and the tidal evolution of star-planet systems. We present a calculation of the fraction of planetary nebulae in the galaxy today which have undergone an interaction with a planet, concluding that a significant number of visible planetary nebulae may have been shaped by a planet.

Contingent-valuation estimates for white-water boating passengers are compared with Likert ratings by river guides. The approach involves asking whether passengers and their guides ordinally rank alternative flows the same. The National Oceanic and Atmospheric Administration's Contingent Valuation Panel (1993) suggested “one might want to compare its (contingent-valuation's) outcome with that provided by a panel of experts.” River guides constitute a counterfactual panel of “experts.” For commercial trips, optimum flows are 34,000 cfs and 31,000 cfs for passengers and guides, and the comparable figures for private trips are 28,000 cfs and 29,000 cfs. In the NOAA Panel framework, passengers can evaluate the consequences of various river flows and translate this into contingent-valuation responses.

Policy makers often face the problem of evaluating how water quality affects a region's economic well-being. Using water clarity as a measure of the degree of eutrophication levels (as a lake becomes inundated with nutrients, water clarity decreases markedly), analysis is performed on sales data collected over a six-year period. Our results indicate that water clarity has a significant effect on prices paid for residential properties. Effects of a one-meter change in clarity on property value are also estimated for an average lake in four real estate market areas in New Hampshire, with effects differing substantially by area. Our findings provide state and local policy makers a measure of the cost of water quality degradation as measured by changes in water clarity, and demonstrate that protecting water quality may have a positive effect on property tax revenues.

Approximately half of the variation in wellbeing measures overlaps with variation in personality traits. Studies of non-human primate pedigrees and human twins suggest that this is due to common genetic influences. We tested whether personality polygenic scores for the NEO Five-Factor Inventory (NEO-FFI) domains and for item response theory (IRT) derived extraversion and neuroticism scores predict variance in wellbeing measures. Polygenic scores were based on published genome-wide association (GWA) results in over 17,000 individuals for the NEO-FFI and in over 63,000 for the IRT extraversion and neuroticism traits. The NEO-FFI polygenic scores were used to predict life satisfaction in 7 cohorts, positive affect in 12 cohorts, and general wellbeing in 1 cohort (maximal N = 46,508). Meta-analysis of these results showed no significant association between NEO-FFI personality polygenic scores and the wellbeing measures. IRT extraversion and neuroticism polygenic scores were used to predict life satisfaction and positive affect in almost 37,000 individuals from UK Biobank. Significant positive associations (effect sizes <0.05%) were observed between the extraversion polygenic score and wellbeing measures, and a negative association was observed between the polygenic neuroticism score and life satisfaction. Furthermore, using GWA data, genetic correlations of -0.49 and -0.55 were estimated between neuroticism with life satisfaction and positive affect, respectively. The moderate genetic correlation between neuroticism and wellbeing is in line with twin research showing that genetic influences on wellbeing are also shared with other independent personality domains.

Immunoglobulin A (IgA) is a predominant immunoglobulin present in human breast milk and is known to play an important role in infant gut immunity maturation. Breast milk composition varies between populations, but the environmental and maternal factors responsible for these variations are still unclear. We examined the relationship between different exposures and levels of IgA in colostrum. The objective of this study was to examine whether exposures analysed influence levels of IgA in colostrum. The present study used 294 colostrum samples from the MecMilk International cohort, collected from women residing in London, Moscow and Verona. Samples were analysed in automated Abbott Architect Analyser. We found an inverse correlation between time postpartum and colostrum total IgA level (r=−0.49, P<0.001). Adjusting for maternal parity, smoking, fresh fruit and fish consumption and allergen sensitization, multiple regression model showed that IgA levels were influenced by colostrum collection time (P<0.0001) and country of collection (P<0.01). Mode of delivery influence did not appear to be significant in univariate comparisons, once adjusted for the above maternal characteristics it showed a significant influence on total IgA (P=0.01). We conclude that the concentration of IgA in colostrum drops rapidly after birth and future studies should always consider this factor in analysis. IgA concentration varied significantly between countries, with the highest level detected in Moscow and lowest in Verona. Mode of delivery effect should be confirmed on larger cohorts. Further work is needed to determine ways to correct for IgA decline over time in colostrum, and to find the cause of variations in IgA levels between the countries.

This paper describes the system architecture of a newly constructed radio telescope – the Boolardy engineering test array, which is a prototype of the Australian square kilometre array pathfinder telescope. Phased array feed technology is used to form multiple simultaneous beams per antenna, providing astronomers with unprecedented survey speed. The test array described here is a six-antenna interferometer, fitted with prototype signal processing hardware capable of forming at least nine dual-polarisation beams simultaneously, allowing several square degrees to be imaged in a single pointed observation. The main purpose of the test array is to develop beamforming and wide-field calibration methods for use with the full telescope, but it will also be capable of limited early science demonstrations.

The Critically Endangered tamaraw Bubalus mindorensis is endemic to Mindoro Island, Philippines, and little is known of its ecology. During 2006–2011 we used community-based monitoring to examine the population status and ecology of tamaraw in the species’ core habitat of Mount Iglit–Baco National Park. Each year, for 5 consecutive days at the end of the dry season, trained local volunteers and rangers or project staff were allocated to 18 vantage points in the study area (c. 160 km2). Tamaraw were categorized as adult (> 5 years), juvenile (2–5 years) or calf (< 2 years), and sexed when possible. During the study period the population was 239–314 (mean 271), with no significant fluctuations in age structure (percentage of adults, juveniles and calves: 57.8, 21.0 and 21.3%) or estimated adult female reproductive rates (29.1%). In adults, but not in juveniles, the sex ratio was biased towards females (1 : 1.86, P < 0.01). Bulls were often solitary (32.2% of sightings), whereas the majority of cows (94.7%) formed small groups of 2–12 individuals of different ages, with or without bulls (53.4 and 46.6%, respectively). These results demonstrate that the population remained relatively stable, maintaining a constant age structure and reproductive rate, and that long-term community-based monitoring was effective for quantitative characterization of the tamaraw's social behaviour, which is critical for conservation and management of the species.

The new photometric observations of PG 2303+243 were obtained in 2012 during a campaign carried out with three telescopes. The analysis of these observations is presented in this paper. We identified l = 1 and l = 2 pulsation modes. The pulsation periods were compared with theoretical ones for models of ZZ Ceti stars. This allowed us to estimate the physical parameters of PG 2303+243. The star seems to be cooler and has thicker hydrogen layer than it was thought before. We have derived M*/M⊙ = 0.66, Teff = 11014 K and log(MH/M*)=−4.246 for this star.