Conducting qualitative interviews in-person is usually presented as the gold standard, with other modes being seen as inferior. There have been arguments, however, that remote interviews, such as ...those conducted using the telephone or videoconference technologies, should be seen as equivalent to or even superior to in-person interviews. Evaluations of these claims have been limited by the small number of interviews used to compare modes. We analyze over 300 interviews conducted using three modes: in-person, telephone, and Skype. Our analyses find that in-person interviews have clear advantages when it comes to producing conversation turns and word-dense transcripts and field notes but do not significantly differ from the other two modes in interview length in minutes, subjective interviewer ratings, and substantive coding. We conclude that, although remote interviews might be necessary or advantageous in some situations, they likely do often come at a cost to the richness of information produced by the interviews.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a beta-CoV that recently emerged as a human pathogen and is the causative agent of the COVID-19 pandemic. A molecular framework of how ...the virus manipulates host cellular machinery to facilitate infection remains unclear. Here, we focus on SARS-CoV-2 NSP1, which is proposed to be a virulence factor that inhibits protein synthesis by directly binding the human ribosome. We demonstrate biochemically that NSP1 inhibits translation of model human and SARS-CoV-2 messenger RNAs (mRNAs). NSP1 specifically binds to the small (40S) ribosomal subunit, which is required for translation inhibition. Using single-molecule fluorescence assays to monitor NSP1-40S subunit binding in real time, we determine that eukaryotic translation initiation factors (eIFs) allosterically modulate the interaction of NSP1 with ribosomal preinitiation complexes in the absence of mRNA. We further elucidate that NSP1 competes with RNA segments downstream of the start codon to bind the 40S subunit and that the protein is unable to associate rapidly with 80S ribosomes assembled on an mRNA. Collectively, our findings support a model where NSP1 proteins from viruses in at least two subgenera of beta-CoVs associate with the open head conformation of the 40S subunit to inhibit an early step of translation, by preventing accommodation of mRNA within the entry channel.
Systemic sclerosis (SSc) is a rare autoimmune connective tissue disease with multi-organ involvement, fibrosis and vasculopathy. Treatment in SSc, including early diffuse cutaneous SSc (dcSSc) and ...the use of organ-specific therapies, has improved, as evident from randomized clinical trials. Treatments for early dcSSc include immunosuppressive agents such as mycophenolate mofetil, methotrexate, cyclophosphamide, rituximab and tocilizumab. Patients with rapidly progressive early dcSSc might be eligible for autologous haematopoietic stem cell transplantation, which can improve survival. Morbidity from interstitial lung disease and pulmonary arterial hypertension is improving with the use of proven therapies. Mycophenolate mofetil has surpassed cyclophosphamide as the initial treatment for SSc-interstitial lung disease. Nintedanib and possibly perfinidone can be considered in SSc pulmonary fibrosis. Pulmonary arterial hypertension is frequently treated with initial combination therapy (for example, with phosphodiesterase 5 inhibitors and endothelin receptor antagonists) and, if necessary, the addition of a prostacyclin analogue. Raynaud phenomenon and digital ulcers are treated with dihydropyridine calcium channel blockers (especially nifedipine), then phosphodiesterase 5 inhibitors or intravenous iloprost. Bosentan can reduce the development of new digital ulcers. Trial data for other manifestations are mostly lacking. Research is needed to develop targeted and highly effective treatments, best practices for organ-specific screening and early intervention, and sensitive outcome measurements.
Advanced LIGO's recent observations of gravitational waves (GWs) from merging binary black holes have opened up a unique laboratory to test general relativity (GR) in the highly relativistic regime. ...One of the tests used to establish the consistency of the first LIGO event with a binary black hole merger predicted by GR was the inspiral-merger-ringdown consistency test. This involves inferring the mass and spin of the remnant black hole from the inspiral (low-frequency) part of the observed signal and checking for the consistency of the inferred parameters with the same estimated from the post-inspiral (high-frequency) part of the signal. Based on the observed rate of binary black hole mergers, we expect the advanced GW observatories to observe hundreds of binary black hole mergers every year when operating at their design sensitivities, most of them with modest signal to noise ratios (SNRs). Anticipating such observations, this paper shows how constraints from a large number of events with modest SNRs can be combined to produce strong constraints on deviations from GR. Using kludge modified GR waveforms, we demonstrate how this test could identify certain types of deviations from GR if such deviations are present in the signal waveforms. We also study the robustness of this test against reasonable variations of a variety of different analysis parameters.
•Leakages through shadow banks associated with prudential capital controls.•Capital outflow controls result in a welfare loss in the policy source country.•Capital inflow controls result in a welfare ...gain in the policy source country.•International transmission of shocks dependent on the type of controls considered.
Shadow banks have recently led the growth in cross-border lending. With this in mind, I develop a two-country real business cycle model with commercial and shadow banking to study the business cycle implications and welfare effects of prudential capital controls. In my model, the presence of shadow banking results in leakages associated with capital controls. As a result, capital outflow restrictions result in a welfare loss in the policy source country, but a welfare improvement in the rival country. Capital inflow restrictions yield the opposite welfare effects for both countries.
The mechanisms that drive nonalcoholic fatty liver disease (NAFLD) remain incompletely understood. This large multicenter study characterized the transcriptional changes that occur in liver tissue ...across the NAFLD spectrum as disease progresses to cirrhosis to identify potential circulating markers. We performed high-throughput RNA sequencing on a discovery cohort comprising histologically characterized NAFLD samples from 206 patients. Unsupervised clustering stratified NAFLD on the basis of disease activity and fibrosis stage with differences in age, aspartate aminotransferase (AST), type 2 diabetes mellitus, and carriage of
, a genetic variant associated with NAFLD. Relative to early disease, we consistently identified 25 differentially expressed genes as fibrosing steatohepatitis progressed through stages F2 to F4. This 25-gene signature was independently validated by logistic modeling in a separate replication cohort (
= 175), and an integrative analysis with publicly available single-cell RNA sequencing data elucidated the likely relative contribution of specific intrahepatic cell populations. Translating these findings to the protein level, SomaScan analysis in more than 300 NAFLD serum samples confirmed that circulating concentrations of proteins AKR1B10 and GDF15 were strongly associated with disease activity and fibrosis stage. Supporting the biological plausibility of these data, in vitro functional studies determined that endoplasmic reticulum stress up-regulated expression of
,
, and
, whereas GDF15 supplementation tempered the inflammatory response in macrophages upon lipid loading and lipopolysaccharide stimulation. This study provides insights into the pathophysiology of progressive fibrosing steatohepatitis, and proof of principle that transcriptomic changes represent potentially tractable and clinically relevant markers of disease progression.
Thrombosis and biofouling of extracorporeal circuits and indwelling medical devices cause significant morbidity and mortality worldwide. We apply a bioinspired, omniphobic coating to tubing and ...catheters and show that it completely repels blood and suppresses biofilm formation. The coating is a covalently tethered, flexible molecular layer of perfluorocarbon, which holds a thin liquid film of medical-grade perfluorocarbon on the surface. This coating prevents fibrin attachment, reduces platelet adhesion and activation, suppresses biofilm formation and is stable under blood flow in vitro. Surface-coated medical-grade tubing and catheters, assembled into arteriovenous shunts and implanted in pigs, remain patent for at least 8 h without anticoagulation. This surface-coating technology could reduce the use of anticoagulants in patients and help to prevent thrombotic occlusion and biofouling of medical devices.
NOX2 is the prototypical member of the NADPH oxidase NOX superfamily and produces superoxide (O
), a key reactive oxygen species (ROS) that is essential in innate and adaptive immunity. Mutations ...that lead to deficiency in NOX2 activity correlate with increased susceptibility to bacterial and fungal infections, resulting in chronic granulomatous disease. The core of NOX2 is formed by a heterodimeric transmembrane complex composed of NOX2 (formerly gp91) and p22, but a detailed description of its structural architecture is lacking. Here, we present the structure of the human NOX2 core complex bound to a selective anti-NOX2 antibody fragment. The core complex reveals an intricate extracellular topology of NOX2, a four-transmembrane fold of the p22 subunit, and an extensive transmembrane interface which provides insights into NOX2 assembly and activation. Functional assays uncover an inhibitory activity of the 7G5 antibody mediated by internalization-dependent and internalization-independent mechanisms. Overall, our results provide insights into the NOX2 core complex architecture, disease-causing mutations, and potential avenues for selective NOX2 pharmacological modulation.
The stability and longevity of surface-stabilized lubricant layers is a critical question in their application as low- and nonfouling slippery surface treatments in both industry and medicine. Here, ...we investigate lubricant loss from surfaces under flow in water using both quantitative analysis and visualization, testing the effects of underlying surface type (nanostructured versus flat), as well as flow rate in the physiologically relevant range, lubricant type, and time. We find lubricant losses on the order of only ng/cm2 in a closed system, indicating that these interfaces are relatively stable under the flow conditions tested. No notable differences emerged between surface type, flow rate, lubricant type, or time. However, exposure of the lubricant layers to an air/water interface did significantly increase the amount of lubricant removed from the surface, leading to disruption of the layer. These results may help in the development and design of materials using surface-immobilized lubricant interfaces for repellency under flow conditions.
Summary
We characterized a set of Arabidopsis mutants deficient in specific light‐harvesting proteins, using freeze‐fracture electron microscopy to probe the organization of complexes in the membrane ...and confocal fluorescence recovery after photobleaching to probe the dynamics of thylakoid membranes within intact chloroplasts. The same methods were used to characterize mutants lacking or over‐expressing PsbS, a protein related to light‐harvesting complexes that appears to play a role in regulation of photosynthetic light harvesting. We found that changes in the complement of light‐harvesting complexes and PsbS have striking effects on the photosystem II macrostructure, and that these effects correlate with changes in the mobility of chlorophyll proteins within the thylakoid membrane. The mobility of chlorophyll proteins was found to correlate with the extent of photoprotective non‐photochemical quenching, consistent with the idea that non‐photochemical quenching involves extensive re‐organization of complexes in the membrane. We suggest that a key feature of the physiological function of PsbS is to decrease the formation of ordered semi‐crystalline arrays of photosystem II in the low‐light state. Thus the presence of PsbS leads to an increase in the fluidity of the membrane, accelerating the re‐organization of the photosystem II macrostructure that is necessary for induction of non‐photochemical quenching.