Viral RNA-host protein interactions are critical for replication of flaviviruses, a genus of positive-strand RNA viruses comprising major vector-borne human pathogens including dengue viruses (DENV). ...We examined three conserved host RNA-binding proteins (RBPs) G3BP1, G3BP2 and CAPRIN1 in dengue virus (DENV-2) infection and found them to be novel regulators of the interferon (IFN) response against DENV-2. The three RBPs were required for the accumulation of the protein products of several interferon stimulated genes (ISGs), and for efficient translation of PKR and IFITM2 mRNAs. This identifies G3BP1, G3BP2 and CAPRIN1 as novel regulators of the antiviral state. Their antiviral activity was antagonized by the abundant DENV-2 non-coding subgenomic flaviviral RNA (sfRNA), which bound to G3BP1, G3BP2 and CAPRIN1, inhibited their activity and lead to profound inhibition of ISG mRNA translation. This work describes a new and unexpected level of regulation for interferon stimulated gene expression and presents the first mechanism of action for an sfRNA as a molecular sponge of anti-viral effectors in human cells.
Extracellular vesicles (EVs) are nano-sized vesicles containing nucleic acid and protein cargo that are released from a multitude of cell types and have gained significant interest as potential ...diagnostic biomarkers. Human serum is a rich source of readily accessible EVs; however, the separation of EVs from serum proteins and non-EV lipid particles represents a considerable challenge. In this study, we compared the most commonly used isolation techniques, either alone or in combination, for the isolation of EVs from 200 µl of human serum and their separation from non-EV protein and lipid particles present in serum. The size and yield of particles isolated by each method was determined by nanoparticle tracking analysis, with the variation in particle size distribution being used to determine the relative impact of lipoproteins and protein aggregates on the isolated EV population. Purification of EVs from soluble protein was determined by calculating the ratio of EV particle count to protein concentration. Finally, lipoprotein particles co-isolated with EVs was determined by Western blot analysis of lipoprotein markers APOB and APOE. Overall, this study reveals that the choice of EV isolation procedure significantly impacts EV yield from human serum, together with the presence of lipoprotein and protein contaminants.
Computational prediction of the behavior of concentrated protein solutions is particularly advantageous in early development stages of biotherapeutics when material availability is limited and a ...large set of formulation conditions needs to be explored. This review provides an overview of the different computational paradigms that have been successfully used in modeling undesirable physical behaviors of protein solutions with a particular emphasis on high-concentration drug formulations. This includes models ranging from all-atom simulations, coarse-grained representations to macro-scale mathematical descriptions used to study physical instability phenomena of protein solutions such as aggregation, elevated viscosity, and phase separation. These models are compared and summarized in the context of the physical processes and their underlying assumptions and limitations. A detailed analysis is also given for identifying protein interaction processes that are explicitly or implicitly considered in the different modeling approaches and particularly their relations to various formulation parameters. Lastly, many of the shortcomings of existing computational models are discussed, providing perspectives and possible directions toward an efficient computational framework for designing effective protein formulations.
Context. To calibrate automatic pipelines that determine atmospheric parameters of stars, one needs a sample of stars, or “benchmark stars”, with well-defined parameters to be used as a reference. ...Aims. We provide detailed documentation of the iron abundance determination of the 34 FGK-type benchmark stars that are selected to be the pillars for calibration of the one billion Gaia stars. They cover a wide range of temperatures, surface gravities, and metallicities. Methods. Up to seven different methods were used to analyze an observed spectral library of high resolutions and high signal-to-noise ratios. The metallicity was determined by assuming a value of effective temperature and surface gravity obtained from fundamental relations; that is, these parameters were known a priori and independently from the spectra. Results. We present a set of metallicity values obtained in a homogeneous way for our sample of benchmark stars. In addition to this value, we provide detailed documentation of the associated uncertainties. Finally, we report a value of the metallicity of the cool giant ψ Phe for the first time.
We consider the structure of the lattice of (order and algebra) ideals of the band of regular kernel operators on
L
p
-spaces. We show, in particular, that for any
L
p
(
μ
)
space, with
μ
σ
-finite ...and
1
<
p
<
∞
, the norm-closure of the ideal of finite-rank operators on
L
p
(
μ
)
, is the only non-trivial proper closed (order and algebra) ideal of this band. Key to our results in the
L
p
setting is the fact that every regular kernel operator on an
L
p
(
μ
)
space (
μ
and
p
as before) factors with regular factors through
ℓ
p
. We show that a similar but weaker factorization property, where
ℓ
p
is replaced by some reflexive purely atomic Banach lattice, characterizes the regular kernel operators from a reflexive Banach lattice with weak order unit to a KB-space with weak order unit.
Virus neutralization remains the gold standard for determining antibody efficacy. Therefore, a high-throughput assay to measure SARS-CoV-2 neutralizing antibodies is urgently needed for COVID-19 ...serodiagnosis, convalescent plasma therapy, and vaccine development. Here, we report on a fluorescence-based SARS-CoV-2 neutralization assay that detects SARS-CoV-2 neutralizing antibodies in COVID-19 patient specimens and yields comparable results to plaque reduction neutralizing assay, the gold standard of serological testing. The fluorescence-based neutralization assay is specific to measure COVID-19 neutralizing antibodies without cross reacting with patient specimens with other viral, bacterial, or parasitic infections. Collectively, our approach offers a rapid platform that can be scaled to screen people for antibody protection from COVID-19, a key parameter necessary to safely reopen local communities.
Objective
To describe the pharmacological management of borderline personality disorder (BPD) in Spain from 2001 through 2016, the factors associated with prescriptions, and changes in ...pharmacotherapy over this time period.
Methods
Retrospective, cross‐sectional, observational study conducted in a sample of 457 patients with BPD consecutively admitted to a specialist BPD Program between January 2001 and November 2016. Data on sociodemographic and clinical variables, as well as pharmacological treatment upon the admission to the programme, were used to describe pharmacological prescriptions, the factors associated with these medications, and changes in prescription over the last 15 years.
Results
Most (88.4%) patients were on pharmacological treatment, with 53.8% of persons taking ≥3 medications. No significant changes in these percentages were observed over the study period. The use of tricyclic antidepressants and benzodiazepines decreased, while the use of atypical antipsychotics increased. Axis I comorbidity was the main factor associated with pharmacological treatment and polypharmacy.
Conclusions
This study provides further evidence confirming the worldwide overuse of prescription medications for BPD and shows that there has been a shift in the prescription pattern in the last 15 years. These results suggest that real clinical practice only partially adheres to clinical treatment guidelines.
While the fundamental series of 2+2cycloadditions and retro2+2cycloadditions that make up the pathways of ruthenium-catalysed metathesis reactions is well-established, the exploration of mechanistic ...aspects of alkene metathesis continues. In this Feature Article, modern mechanistic studies of the alkene metathesis reaction, catalysed by well-defined ruthenium complexes, are discussed. Broadly, these concern the processes of pre-catalyst initiation, propagation and decomposition, which all have a considerable impact on the overall efficiency of metathesis reactions.
While the fundamental series of 2+2cycloadditions and retro2+2cycloadditions that make up the pathways of ruthenium-catalysed metathesis reactions is well-established, the exploration of mechanistic aspects of alkene metathesis continues.
Using a multi-camera computerised tracking system the present study aimed to provide a detailed analysis of the work-rate profile of a team of elite soccer players during official matches of the ...Spanish Premier League. Observation-based performance measures were obtained from 434 individual samples. 6 physical parameters involving the distance covered by players were analysed: standing intensity (0-11 km·h (-1)), low-intensity running (11.1-14 km·h (-1)), moderate-intensity running (14.1-17 km·h (-1)), high-intensity running (17.1-21 km·h (-1)), very high-intensity running (21.1-24 km·h (-1)) and sprinting (>24 km·h (-1)). These intensity thresholds were considered with respect to 4 contextual variables: MATCH STATUS, MATCH LOCATION, OPPONENT LEVEL and MATCH HALF, which were analysed in relation to the EFFECTIVE PLAYING TIME. A descriptive analysis and a multivariate mixed model were employed for the analysis of change processes in soccer. The distance total covered (m) by players at different work intensities during the EFFECTIVE PLAYING TIME was greater when playing at HOME (3 931 vs. 3 887 AWAY), when the reference team was LOSING (3 975 vs. 3 837 DRAWING and 3 921 WINNING) and when the level of the opposing team was HIGHER (4 032 vs. 3 938 MEDIUM and 3 736 BOTTOM). By contrast, their physical performance decreased during the 2NDHALF of matches (3 822 vs. 3 985 1ST HALF).
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