Kinetic analysis of RNA cleavage by coronavirus Nsp15 endonuclease: Evidence for acid-base catalysis and substrate-dependent metal ion activation.

Understanding the functional properties of severe acute respiratory syndrome coronavirus 2 nonstructural proteins is essential for defining their roles in the viral life cycle, developing improved therapeutics and diagnostics, and countering future variants. Coronavirus nonstructural protein Nsp15 is a hexameric U-specific endonuclease whose functions, substrate specificity, mechanism, and dynamics are not fully defined. Previous studies report that Nsp15 requires Mn2+ ions for optimal activity; however, the effects of divalent ions on Nsp15 reaction kinetics have not been investigated in detail. Here, we analyzed the single- and multiple-turnover kinetics for model ssRNA substrates. Our data confirm that divalent ions are dispensable for catalysis and show that Mn2+ activates Nsp15 cleavage of two different ssRNA oligonucleotide substrates but not a dinucleotide. Biphasic kinetics of ssRNA substrates demonstrates that Mn2+ stabilizes alternative enzyme states that have faster substrate cleavage on the enzyme. However, we did not detect Mn2+-induced conformational changes using CD and fluorescence spectroscopy. The pH-rate profiles in the presence and absence of Mn2+ reveal active-site ionizable groups with similar pKas of ca. 4.8 to 5.2. An Rp stereoisomer phosphorothioate modification at the scissile phosphate had minimal effect on catalysis supporting a mechanism involving an anionic transition state. However, the Sp stereoisomer is inactive because of weak binding, consistent with models that position the nonbridging phosphoryl oxygen deep in the active site. Together, these data demonstrate that Nsp15 employs a conventional acid-base catalytic mechanism passing through an anionic transition state, and that divalent ion activation is substrate dependent.

A behavioral insights approach to recruiting entrepreneurs for an academic study during the COVID-19 pandemic

What should researchers say when recruiting entrepreneurs to participate in their study? Using a sample of entrepreneurs (N = 1,450) who were being asked to participate in an academic research project, we conducted an experiment to determine recruitment message efficacy. Drawing on best practices from the behavioral insights literature, we developed different email message recruitment statements that were randomly assigned across four phases of our experiment. Results indicate that a message grounded in the "descriptive norms” (i.e., social norms) approach resulted in the highest percentage of participants who clicked on the link to participate in our online survey. We discuss the theoretical as well as practical implications of our work.

Beyond forced telehealth adoption: A framework to sustain telehealth among allied health services.

INTRODUCTION: As COVID-19 restrictions reduce globally, services will determine what components of care will continue via telehealth. We aimed to determine the clinician, service, and system level factors that influence sustained use of telehealth and develop a framework to enhance sustained use where appropriate. METHODS: This study was conducted across 16 allied health departments over four health service facilities (Brisbane, Australia). It used a multi-method observational study design, involving telehealth service activity data from hospital administrative databases and qualitative interviews with allied health staff (n = 80). Data were integrated and analysed using Greenhalgh's Non-adoption, Abandonment, Scale-up, Spread, and Sustainability framework. RESULTS: Increased telehealth use during the peak COVID period reverted to in-person activity as restrictions eased. Telehealth is unlikely to be sustained without a clear strategy including determination of roles and responsibilities across the organisation. Clinician resistance due to forced adoption remains a key issue. The main motivator for clinicians to use telehealth was improved consumer-centred care. Benefits beyond this are needed to sustain telehealth and improvements are required to make the telehealth experience seamless for providers and recipients. Data were synthesised into a comprehensive framework that can be used as a blueprint for system-wide improvements and service enhancement or redesign. DISCUSSION: Sustainability of telehealth activity beyond the peak COVID period is unlikely without implementation strategies to address consumer, clinician, service, and system factors. The framework can inform how these strategies can be enacted. Whilst developed for allied health disciplines, it is likely applicable to other disciplines.

An Overview of Rural Entrepreneurship and Future Directions

Prior research shows that rural entrepreneurship has its own distinct elements and deserves additional attention within the research community. The frameworks and methodologies from studies focused on high-growth and technology-based entrepreneurship are often used to explore rural entrepreneurial activities. This incongruence limits our understanding of the true impact entrepreneurship can have on rural communities. The articles in this special issue help advance our knowledge of rural entrepreneurship as a distinct field of study, and add to our understanding of its impact in the rural context. Additional research avenues are suggested.

Jordan's Public Policy Response to COVID-19 Pandemic: Insight and Policy Analysis

The aim of this study was to gain an understanding of how the Jordanian government has responded and continues to respond to the COVID-19 pandemic. It utilizes the interpretive policy analysis approach through document analysis. The analysis showed that Jordan created social protection policies to assist people who lost their jobs or whose work was suspended due to the coronavirus. The economic policies build solidarity and facilitate the private sector’s recovery. The health care measures firmly applied included lockdown, wearing masks, and restrictions on gatherings and public events. Jordan uses hard power and imposes sanctions on any violation that threatens the lives of others.

Men's attitudes toward mask-wearing during COVID-19: Understanding the complexities of mask-ulinity.

Path analyses indicated that the relationship between conformity to masculine norms (CMN) and attitudes toward mask-wearing to protect from COVID-19 was mediated by perceived benefits, perceived barriers, confidence in the scientific community, and empathy toward vulnerable persons, and that political ideology moderated the indirect effects from CMN to men's attitudes. Efforts to improve men's participation in combatting COVID-19 should address the perceptions and attitudes related to conforming to traditional masculine norms and moderated by political ideology, and might reconstruct masculinity to focus on being a guardian and protector of public health.