Structure-Based Discovery of Allosteric Inhibitors Targeting a New Druggable Site in the Respiratory Syncytial Virus Polymerase.

Respiratory syncytial virus (RSV) is a major cause of severe lower respiratory infections for which effective treatment options remain limited. Herein, we employed a computational structure-based design strategy aimed at identifying potential targets for a new class of allosteric inhibitors. Our investigation led to the discovery of a previously undisclosed allosteric binding site within the RSV polymerase, the large (L) protein. This discovery was achieved through a combination of virtual screening and molecular dynamics simulations. Subsequently, we identified two inhibitors, 6a and 10b, which both exhibited promising antiviral activity in the low micromolar range. Resistance profiling revealed a distinctive pattern in how RSV evaded treatment with this class of inhibitors. This pattern strongly suggested that this class of small molecules was targeting a new binding site in the RSV L protein, aligning with the computational predictions made in our study. This study paves the way for the development of more potent inhibitors for combating RSV infections by targeting a new druggable pocket within the RdRp which does not overlap with previously known resistance sites.

The Effects of Astaxanthin on Cognitive Function and Neurodegeneration in Humans: A Critical Review.

Oxidative stress is a key contributing factor in neurodegeneration, cognitive ageing, cognitive decline, and diminished cognitive longevity. Issues stemming from oxidative stress both in relation to cognition and other areas, such as inflammation, skin health, eye health, and general recovery, have been shown to benefit greatly from antioxidant use. Astaxanthin is a potent antioxidant, which has been outlined to be beneficial for cognitive function both in vitro and in vivo. Given the aforementioned promising effects, research into astaxanthin with a focus on cognitive function has recently been extended to human tissue and human populations. The present critical review explores the effects of astaxanthin on cognitive function and neurodegeneration within human populations and samples with the aim of deciphering the merit and credibility of the research findings and subsequently their potential as a basis for therapeutic use. Implications, limitations, and areas for future research development are also discussed. Key findings include the positive impacts of astaxanthin in relation to improving cognitive function, facilitating neuroprotection, and slowing neurodegeneration within given contexts.

Mapping respiratory syncytial virus fusion protein interactions with the receptor IGF1R and the impact of alanine-scanning mutagenesis on viral infection.

Respiratory syncytial virus (RSV) has two main surface glycoproteins, the attachment glycoprotein (G) and the fusion (F) protein, which together mediate viral entry. Attachment is mediated by the RSV-G protein, while the RSV-F protein makes specific contact with the cellular insulin-like growth factor 1 receptor (IGF1R). This interaction leads to IGF1R activation and initiates a signalling cascade that calls the co-receptor, nucleolin, from the nucleus to the cell surface, where it can trigger viral fusion. We performed molecular docking analysis, which provided a potential set of 35 residues in IGF1R that may be important for interactions with RSV-F. We used alanine-scanning mutagenesis to generate IGF1R mutants and assessed their abundance and maturation, as well as the effect of mutation on RSV infection. We identified several mutations that appear to inhibit IGF1R maturation; but surprisingly, these mutations had no significant effect on RSV infection. This suggests that maturation of IGF1R may not be required for RSV infection. Additionally, we identified one residue, S788, that, when mutated, significantly reduced RSV infection. Further analysis revealed that this mutation disrupted a hydrogen bonding network that may be important for both IGF1R maturation and RSV infection.

Loss of Respiratory Syncytial Virus Antibody Functions During the Peak of the COVID-19 Pandemic Mitigation Measures.

Studies have linked respiratory syncytial virus (RSV) antibody-mediated phagocytosis and complement deposition to severe RSV infection in humans. This study shows waning of these antibody functions in women of childbearing age in 2020-2021 during the implementation of COVID-19 mitigation measures, in absence of RSV circulation. These functions could be explored as correlates of protection against severe RSV disease.

Actin cytoskeleton remodeling disrupts physical barriers to infection and presents entry receptors to respiratory syncytial virus.

RSV is the leading cause of infant hospitalizations and a significant cause of paediatric and geriatric morbidity worldwide. Recently, we reported that insulin-like growth factor 1 receptor (IGF1R) was a receptor for respiratory syncytial virus (RSV) in airway epithelial cells and that activation of IGF1R recruited the coreceptor, nucleolin (NCL), to the cell surface. Cilia and mucus that line the airways pose a significant barrier to viral and bacterial infection. The cortical actin cytoskeleton has been shown by others to mediate RSV entry, so we studied the roles of the RSV receptors and actin remodelling during virus entry. We found that IGF1R expression and phosphorylation were associated with the ability of RSV to infect cells. Confocal immunofluorescence imaging showed that actin projections, a hallmark of macropinocytosis, formed around viral particles 30 min after infection. Consistent with prior reports we also found that virus particles were internalized into early endosome antigen-1 positive endosomes within 90 min. Inhibiting actin polymerization significantly reduced viral titre by approximately ten-fold. Inhibiting PI3 kinase and PKCζ in stratified air-liquid interface (ALI) models of the airway epithelium had similar effects on reducing the actin remodelling observed during infection and attenuating viral entry. Actin projections were associated with NCL interacting with RSV particles resting on apical cilia of the ALIs. We conclude that macropinocytosis-like actin projections protrude through normally protective cilia and mucus layers of stratified airway epithelium that helps present the IGF1R receptor and the NCL coreceptor to RSV particles waiting at the surface.

Developing a simple risk metric for the effect of sport-related concussion and physical pain on mental health.

Risk factors associated with depression in athletes include biological sex, physical pain, and history of sport-related concussion (SRC). Due to the well-documented benefits of sport and physical activity on mental health, athletes and non-athletes were recruited to assess any differences. Beyond this, athletes were also grouped by sport-type (contact/non-contact sports) due to the increased prevalence of pain and SRC in contact sports. To our knowledge, there has been no research on how these factors influence the likelihood of depression. In the current study, 144 participants completed a short survey on the above factors and the Center for Epidemiological Studies Depression Scale. Sixty-two of these reported a history of concussion. Logistic regression revealed all the above predictors to be significantly associated with the depression scale. Individuals that had previously sustained SRC, were experiencing greater physical pain and females were more likely to display poor mental health. However, we provide further evidence for the benefits of engaging in sport and physical activity as those that took part in sport were less likely to report depression. Therefore, this study provides a simple risk metric whereby sportspeople can make a better informed choice of their sporting participation, making their own cost/reward judgement.

Special Issue of the 4th Symposium of the Canadian Society for Virology 2022.

In this Special Issue of Viruses, we showcase some of the fascinating and diverse virology being undertaken in Canada that was presented at the 4th Symposium of the Canadian Society for Virology 2022 [...].

The domestication of SARS-CoV-2 into a seasonal infection by viral variants.

Introduction: The COVID-19 pandemic was caused by the zoonotic betacoronavirus SARS-CoV-2. SARS-CoV-2 variants have emerged due to adaptation in humans, shifting SARS-CoV-2 towards an endemic seasonal virus. We have termed this process 'virus domestication'. Methods: We analyzed aggregate COVID-19 data from a publicly funded healthcare system in Canada from March 7, 2020 to November 21, 2022. We graphed surrogate calculations of COVID-19 disease severity and SARS-CoV-2 variant plaque sizes in tissue culture. Results and Discussion: Mutations in SARS-CoV-2 adapt the virus to better infect humans and evade the host immune response, resulting in the emergence of variants with altered pathogenicity. We observed a decrease in COVID-19 disease severity surrogates after the arrival of the Delta variant, coinciding with significantly smaller plaque sizes. Overall, we suggest that SARS-CoV-2 has become more infectious and less virulent through viral domestication. Our findings highlight the importance of SARS-CoV-2 vaccination and help inform public policy on the highest probability outcomes during viral pandemics.

SARS-CoV-2 infection downregulates myocardial ACE2 and potentiates cardiac inflammation in humans and hamsters.

Myocardial pathologies resulting from SARS-CoV-2 infections are consistently rising with mounting case rates and reinfections; however, the precise global burden is largely unknown and will have an unprecedented impact. Understanding the mechanisms of COVID-19-mediated cardiac injury is essential toward the development of cardioprotective agents that are urgently needed. Assessing novel therapeutic strategies to tackle COVID-19 necessitates an animal model that recapitulates human disease. Here, we sought to compare SARS-CoV-2-infected animals with patients with COVID-19 to identify common mechanisms of cardiac injury. Two-month-old hamsters were infected with either the ancestral (D614) or Delta variant (B.1.617.2) of SARS-CoV-2 for 2 days, 7 days, and/or 14 days. We measured viral RNA and cytokine expression at the earlier time points to capture the initial stages of infection in the lung and heart. We assessed myocardial angiotensin-converting enzyme 2 (ACE2), the entry receptor for the SARS-CoV-2 virus, and cardioprotective enzyme, as well as markers for inflammatory cell infiltration in the hamster hearts at days 7 and 14. In parallel, human hearts were stained for ACE2, viral nucleocapsid, and inflammatory cells. Indeed, we identify myocardial ACE2 downregulation and myeloid cell burden as common events in both hamsters and humans infected with SARS-CoV-2, and we propose targeting downstream ACE2 downregulation as a therapeutic avenue that warrants clinical investigation.NEW & NOTEWORTHY Cardiac manifestations of COVID-19 in humans are mirrored in the SARS-CoV-2 hamster model, recapitulating myocardial damage, ACE2 downregulation, and a consistent pattern of immune cell infiltration independent of viral dose and variant. Therefore, the hamster model is a valid approach to study therapeutic strategies for COVID-19-related heart disease.

Effects of attentional focus and cognitive load on novice dart throwing: Evidence from quiet eye duration and pupillary responses.

The purpose of the present study was to investigate the effects of attentional focus and cognitive-load on motor performance, quiet-eye-duration, and pupil dilation. 18 participants completed a dart throwing task under four conditions, internal or external focus with high or low cognitive-load. Cognitive-load was created by a secondary tone detection task. During each trial participants pupil size and eye movements were recorded along with accuracy data of the dart throw. Results revealed that decreased cognitive-load increased accuracy while high load increased pupil size (p's < 0.05). An external focus resulted in the greatest accuracy while an external focus with high cognitive-load resulted in the longest quiet-eye-durations (p's < 0.05). Based on these findings an increase in pupil size is related to greater cognitive-load but doesn't explain the improvement in task performance. Likewise, an external focus of attention improved performance but was not strongly related to quiet-eye-duration. Results are further discussed in the article.

Self-Paced Endurance Performance and Cerebral Hemodynamics of the Prefrontal Cortex: A Scoping Review of Methodology and Findings.

Recent research has suggested that top-down executive function associated with the prefrontal cortex is key to the decision-making processes and pacing of endurance performance. A small but growing body of literature has investigated the neurological underpinnings of these processes by subjecting the prefrontal cortex to functional near-infrared spectroscopy (fNIRS) measurement during self-paced endurance task performance. Given that fNIRS measurement for these purposes is a relatively recent development, the principal aim of this review was to assess the methodological rigor and findings of this body of research. We performed a systematic literature search to collate research assessing prefrontal cortex oxygenation via fNIRS during self-paced endurance performance. A total of 17 studies met the criteria for inclusion. We then extracted information concerning the methodology and findings from the studies reviewed. Promisingly, most of the reviewed studies reported having adopted commonplace and feasible best practice guidelines. However, a lack of adherence to these guidelines was evident in some areas. For instance, there was little evidence of measures to tackle and remove artifacts from data. Lastly, the reviewed studies provide insight into the significance of cerebral oxygenation to endurance performance and the role of the prefrontal cortex in pacing behavior. Therefore, future research that better follows the guidelines presented will help advance our understanding of the role of the brain in endurance performance and aid in the development of techniques to improve or maintain prefrontal cortex (PFC) oxygenation to help bolster endurance performance.

Waning Immunity Against Respiratory Syncytial Virus During the Coronavirus Disease 2019 Pandemic.

Health jurisdictions have seen a near-disappearance of respiratory syncytial virus (RSV) during the first year of the coronavirus disease 2019 (COVID-19) pandemic. Over this corresponding period, we report a reduction in RSV antibody levels and live virus neutralization in sera from women of childbearing age and infants between May to June 2020 and February to June 2021, in British Columbia (BC), Canada. This supports that antibody immunity against RSV is relatively short-lived and that maintaining optimal antibody levels in infants requires repeated maternal viral exposure. Waning immunity may explain the interseasonal resurgence of RSV cases observed in BC and other countries.

A self-determination theory based investigation of life skills development in youth sport.

This study investigated if basic need satisfaction and frustration mediated the associations between autonomy-supportive and controlling coaching behaviours and participants' development of eight different life skills in youth sport. British sports participants (N = 309, Mage = 14.71) completed measures assessing the study variables. Correlational analyses showed that autonomy-supportive coaching behaviours were positively associated with the satisfaction of participants' three basic needs (autonomy, competence, and relatedness) and their development of all eight life skills, whereas controlling coaching behaviours were only positively related to the frustration of participants' three basic needs. Mediational analyses revealed that satisfaction of all three basic needs combined (total need satisfaction) mediated the associations between autonomy-supportive coaching behaviours and participants' development of the eight life skills. Relatedness satisfaction mediated the associations between autonomy-supportive coaching behaviours and participants' development of all eight life skills except for goal setting; autonomy satisfaction mediated the associations between autonomy-supportive coaching behaviours and participants' time management skills; and competence satisfaction mediated the associations between autonomy-supportive coaching behaviours and participants' goal setting and emotional skills. Based on such findings, coaches should look to display autonomy-supportive behaviours that help to satisfy participants' three basic psychological needs and promote their life skills development in sport.

Effects of Attentional Strategies on Novice Dart Throwing, Quiet Eye Duration and Pupillary Responses.

This study examined the effects of focus of attention (FOA) instructions on learning a dart throwing task, quiet eye duration (QED) and pupillary responses. Thirty-six novices (M age = 33.6, SD = 19.7) learned using either (a) internal-focus (arm movement), (b) external-focus (target/dart) or (c) control instructions before completing retention and transfer tests 10 days later. FOA strategies did not significantly benefit dart throwing learning or performance. QED was associated with performance in the transfer task, and pupillary constriction occurred during the QED. The content of FOA instructions may have limited anticipated performance benefits and reduced the FOA influence on QED. Pupillary constriction reflected increased cognitive effort during QED, suggesting an important feature of future precision aiming research.

Imaging Flow Cytometry and Confocal Immunofluorescence Microscopy of Virus-Host Cell Interactions.

Viruses are diverse pathogens that use host factors to enter cells and cause disease. Imaging the entry and replication phases of viruses and their interactions with host factors is key to fully understanding viral infections. This review will discuss how confocal microscopy and imaging flow cytometry are used to investigate virus entry and replication mechanisms in fixed and live cells. Quantification of viral images and the use of cryo-electron microscopy to gather structural information of viruses is also explored. Using imaging to understand how viruses replicate and interact with host factors, we gain insight into cellular processes and identify novel targets to develop antiviral therapeutics and vaccines.

Pharmacological targets and emerging treatments for respiratory syncytial virus bronchiolitis.

RSV infection of the lower respiratory tract in infants is the leading cause of pediatric hospitalizations and second to malaria in causing infant deaths worldwide. RSV also causes substantial morbidity in immunocompromised and elderly populations. The only available therapeutic is a prophylactic drug called Palivizumab that is a humanized monoclonal antibody, given to high-risk infants. However, this intervention is expensive and has a limited impact on annual hospitalization rates caused by RSV. No vaccine is available, nor are efficacious antivirals to treat an active infection, and there is still no consensus on how infants with bronchiolitis should be treated during hospital admission. In this comprehensive review, we briefly outline the function of the RSV proteins and their suitability as therapeutic targets. We then discuss the most promising drug candidates, their inhibitory mechanisms, and whether they are in the process of clinical trials. We also briefly discuss the reasons for some of the failures in RSV therapeutics and vaccines. In summary, we provide insight into current antiviral development and the considerations toward producing licensed antivirals and therapeutics.

The Effects of Acute Moderate and High Intensity Exercise on Memory.

Acute cardiovascular exercise can enhance correct remembering but its impact upon false remembering is less clear. In two experiments, we investigated the effect of acute bouts of exercise on correct and false remembering using the Deese-Roediger-McDermott (DRM) memory test. In Experiment 1, healthy adults completed quiet rest or moderate intensity cycling prior to the memory test. In Experiment 2, a similar sample completed moderate intensity running, high intensity sprints, or a period of quiet rest prior to the memory test. In Experiment 1, acute moderate intensity exercise increased short-term correct, but not false, recall. Experiment 2 replicated these findings but also found an acute bout of high intensity exercise had no impact upon either type of short-term recall. Acute moderate intensity exercise, but not acute high intensity exercise, can improve short-term correct recall without an accompanying increase in false recall potentially through processing of contextually specific information during encoding.

A test of optimal theory on young adolescents' standing long jump performance and motivation.

The OPTIMAL theory of motor learning contends that an external focus of attention (EF), enhanced expectancies (EE), and autonomy support (AS) are key attentional and motivational variables that optimise motor performance. We examined how integrating an EF into EE and AS interventions would impact young adolescents' standing long jump performance and self-efficacy, perceived competence, task effort, task importance and positive affect. Forty-eight participants completed 3 jumps in a baseline, EF (focus on jumping towards the cone), EE-EF (positive social-comparative feedback/high success probability) and AS-EF (self-definition of success) conditions. Both the EF and AS-EF conditions (but not the EE-EF condition) improved jump performance from baseline. The EF, EE-EF and AS-EF conditions improved young adolescents' self-efficacy, perceived competence, task effort and positive affect in comparison to baseline and were predictors of jump performance (as was task importance). However, in the EE-EF condition motivational states improved (from baseline) but this did not translate into performance improvements. The findings show that directing attention to visual external cues both independently and when framed within AS conditions enhanced young adolescents' jump performance and motivation through efficient goal-action coupling. In practice, PE teachers and sports coaches working with young adolescents can support autonomy by allowing self-definition of success using an external cue to enhance effective goal-action coupling, motor performance and motivation.