Discovery of orally active SARS-CoV-2 papain-like protease (PLpro) inhibitors (preprint)

Vaccines and first-generation antiviral therapeutics have provided important protection against coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, there remains a need for additional therapeutic options that provide enhanced efficacy and protection against potential viral resistance. The SARS-CoV-2 papain-like protease (PLpro) is one of two essential cysteine proteases involved in viral replication. While inhibitors of the SARS-CoV-2 main protease (Mpro) have demonstrated clinical efficacy, known PLpro inhibitors have to date lacked the inhibitory potency and requisite pharmacokinetics to demonstrate that targeting PLpro translates to in vivo efficacy in a preclinical setting. Herein, we report the machine learning-driven discovery of potent, selective, and orally available SARS-CoV-2 PLpro inhibitors, with lead compound PF-07957472 (4) providing robust efficacy in a mouse-adapted model of COVID-19 infection.

Comparison of the analytical and clinical sensitivity of thirty-four rapid antigen tests with the most prevalent SARS-CoV-2 variants of concern during the COVID-19 pandemic in the UK (preprint)

Background The continued emergence of SARS-CoV-2 variants of concern (VOC) requires timely analytical and clinical evaluation of antigen-based rapid diagnostic tests (Ag-RDTs) especially those that are recommended for at home use. Methods The limit of detection (LOD) of 34 Ag-RDTs was evaluated using the most encountered SARS-CoV-2 VOC viral isolates (Alpha, Delta, Gamma, Omicron BA.1, Omicron BA.5) and the wild type (WT). Clinical sensitivity was further evaluated for five Ag-RDT utilising retrospective samples (Alpha, Delta, Omicron BA.1) and one Ag-RDT utilising prospective clinical samples (Delta and Omicron BA.1). Findings For the WT, Alpha, Delta, Gamma and Omicron (BA.1) variants 22, 32, 29, 31 and 32 of the 34 Ag-RDTs evaluated met the World Health Organisations (WHO) target product profile (TPP), respectively. Of the 31 Ag-RDTs included for Omicron BA.5 evaluation 29 met the WHO TPP. Additionally, the LODs for samples spiked with Omicron BA.5 were significantly lower than all other VOCs included (p<0.001). In the retrospective clinical evaluation when comparing RNA copies/mL, the Ag-RDTs detected Alpha and Omicron (BA.1) more sensitively than the Delta VOC. Samples with high RT-qPCR Cts (Ct>25) resulted in reduced test sensitivities across all variants. We used linear regression to model the 50% and 95% LOD of clinical samples and observed statistically similar results for all tests. In the prospective clinical samples, the sensitivity was statistically similar for the Delta VOC 71.9% (CI 95% 53.3-86.6%) and Omicron VOC 84.4% (CI95% 75.3-91.2%). Interpretation Test performance differs between SARS-CoV-2 VOCs, and high sensitivity was achieved when testing the Omicron BA.5 VOC compared to the WHO Ag-RDT requirements. Continuous evaluations must be performed to monitor test performance.

Developing an aid for virtual clinics for education and identification of lipohypertrophy during the Covid-19 pandemic: Diabetesclinic@ home

Aims/objectives: To develop an app to support virtual diabetes clinics to help people with diabetes to check themselves for lipos and to discuss what they found during the virtual consultation. Method(s): Collaborative project between clinical teams and medical illustrators in Swansea Bay University Health Board, academics in Swansea University and Eli Lilly under a Collaborative Working Agreement. The teams worked together to develop the content, animations, and a learning technologist developed progressive web app (PWA). The app was tested by diabetes patient groups in Wales, as well as HCP groups, Welsh Academy for Nurses in Diabetes (WAND) and Diabetes Specialist Nurses (DSN) forum prior to launch in June 2021. Result(s): Between 28th June 2021 and 11th November 2022 the app had 827 unique users and 124 returning users. Users came from 15 different countries including UK, Australia, USA, Germany, Brazil and Saudi Arabia. The proportion of UK users were 480 (60%) England, 265 (34%) Wales, and 19 (2%) Scotland. A total of 41 users completed the feedback form;of those 11 (27%) did not know about lipos prior to using the app, 11 (27%) found a lipo using the app, 14 (34%) said they made changes to injection technique after using the app and 37 (90%) said their knowledge of lipos increased following using the app. Conclusion(s): A PWA can help to aid virtual clinics an provide education for people with diabetes. The diabetesclinic@ home app improved knowledge and detection of lipos and improved injection technique.

Impact of the COVID-19 Pandemic on Cervical Cancer Screening Rates in Colombia: An Interrupted Time Series Analysis, Stratified By Region

Objectives: The objective of this study is to measure the national impact of COVID-19 on cervical cancer screening rates in Colombia in five of its geographic regions to inform future health policy decision making. Method(s): This study utilized a quasi-experimental interrupted time-series design to examine changes in trends for the number of cervical cancer screenings performed in five geographic regions of Colombia. Result(s): In the rural region of Vichada, we found the lowest incidence of cervical cancer screenings, totaling at 3,771 screenings. In Cundinamarca, the region which hosts the capital city, a total of 1,213,048 cervical cancer screenings were performed. The researcher measured the impact on cervical cancer screenings in December 2021 against the counterfactual. This impact was ~269 cases that were not performed in December 2021 as a result of the COVID-19 pandemic compared to the counterfactual. In Cundinamarca, unlike other regions, we observed a stagnant pre-pandemic trend, a sharp drop in screenings in March 2020, and an immediate upward trend starting in April 2020. In the month of April 2020, compared to the counterfactual, there were 27,359 screenings missed, and by the month of December 2021, there were only 5,633 cervical cancer screenings missed. Conclusion(s): The region of Cundinamarca's sharp climb back to pre-pandemic screening levels could signal the relatively stronger communication system in the region, and especially in the capital district of Bogota, in re-activating the economy. This can serve as an example of what should be implemented in other regions to improve cervical cancer screening rates. Areas for further research include the examination of social determinants of health, such as the breakdown of the type of insurance screened patients hold (public versus private), zone (urban versus rural), insurance providers of those screened, ethnicities of the patients screened, and percentage of screenings that resulted in early detection of cervical cancer.Copyright © 2023

Relationships Between Burnout and Resilience: Experiences of Physical Therapists and Occupational Therapists During the COVID-19 Pandemic.

OBJECTIVE: Research on burnout among physical therapists and occupational therapists in the context of the coronavirus disease 2019 (COVID-19) pandemic is limited. Resilience may be important for reducing burnout and promoting well-being among rehabilitation specialists, especially during periods of elevated occupational demand and stress. The purpose of this study was to investigate experiences of burnout, COVID-19 pandemic-related distress, and resilience among physical therapists and occupational therapists during the first year of the COVID-19 pandemic. METHODS: Physical therapists and occupational therapists working in a university-affiliated health system were invited to complete an online survey assessing burnout, COVID-19 pandemic-related distress, state- and trait-like resilience, physical activity, sleep disturbance, and financial concerns. Multiple linear regressions were used to examine variables associated with burnout as well as the contribution of specific aspects of resilience to burnout. RESULTS: Greater COVID-19 pandemic-related distress was associated with greater emotional exhaustion and depersonalization, whereas state-like resilience at work was associated with lower emotional exhaustion, greater personal accomplishment, and lower depersonalization. Analyses examining the impact of specific components of resilience at work suggested that several components are associated with less burnout, with finding one's calling being particularly relevant for all 3 domains of burnout. CONCLUSION: Symptoms of burnout were reported by many physical therapists and occupational therapists. COVID-19-related distress and state-like resilience at work, particularly the perception of finding one's calling, emerged as consistently being associated with burnout in the context of the COVID-19 pandemic. IMPACT: These findings can inform the development of interventions to reduce burnout among physical therapists and occupational therapists amid the continuing COVID-19 pandemic.

Understanding First Contact Physiotherapists Experiences of Remote Consultations in Primary Care

Background/Aims First Contact Physiotherapists (FCPs) assess, diagnose and manage patients presenting with musculoskeletal disorders in primary care, without the need for prior GP consultation. Prior to COVID-19 almost every consultation was conducted in-person. Since the pandemic, many consultations are now undertaken remotely, a trend that is set to continue in line with the 'Digital first' strategy which seeks to enhance patient access to appointments. This aim of this study was to explore FCP views of remote consultations and how this impacted their role satisfaction and wellbeing. Methods This mixed methods two phase study consisted of an online survey investigating distributed via professional networks and through social media. The phase one survey explored consultation methods;levels of training;challenges and benefits;and a stress appraisal. Data were analysed descriptively. Respondents were invited to take part in phase two which included a semi-structured interview to gain an in-depth understanding of FCPs lived experience of remote consultation ways of working. Transcripts were thematically analysed. Results The online survey received n=109 responses from UK-based FCPs. Data revealed that despite the 'Digital First' push for continued remote consultations, the majority of FCPs (62%) used them for less than a quarter of their appointment slots. Whilst recognising that many patients found this format convenient, FCPs highlighted their own stress levels, citing poor efficacy, anxiety of misdiagnosis, feelings of isolation and increased administrative workload. Nearly two thirds (66%) of respondents had not received any training in how to conduct effective remote consultations. Follow-up interviews with n=16 FCPs highlighted coping strategies including following up with an in-person consultation and directing patients to other community health and wellbeing resources. In areas of high socioeconomic deprivation and poor health literacy additional problems associated with communication difficulties, poor IT access and capability, and digital poverty were all cited. Conclusion Remote consultations may offer a convenient alternative for some patients. FCP responses suggest that the continued offer of remote consultation is decreasing now pandemic restrictions have been lifted, despite the push for continued digital working practices. The perceived lack of efficacy, and fear of missing important diagnostic information means that many FCPs are either returning to in-person consultation or following up with a second face-to-face assessment resulting in potential service inefficiencies. Additional challenges were identified in areas of high deprivation and low health literacy, and the value of this consultation format needs to be considered in this context. Future work should focus on the training and support needs of FCP staff who are engaging with remote working to ensure clinical effectiveness and staff wellbeing.

Lipase Elevation in Patients With COVID-19.

INTRODUCTION: Although coronavirus disease (COVID-19) has been associated with gastrointestinal manifestations, its effect on the pancreas remains unclear. We aimed to assess the frequency and characteristics of hyperlipasemia in patients with COVID-19. METHODS: A retrospective cohort study of hospitalized patients across 6 US centers with COVID-19. RESULTS: Of 71 patients, 9 (12.1%) developed hyperlipasemia, with 2 (2.8%) greater than 3 times upper limit of normal. No patient developed acute pancreatitis. Hyperlipasemia was not associated with poor outcomes or symptoms. DISCUSSION: Although a mild elevation in serum lipase was observed in some patients with COVID-19, clinical acute pancreatitis was not seen.

Unique Differentiation and Homing Features of T Cells in Individuals with Long Covid

Background: A significant portion of individuals experience persistent symptoms months after SARS-CoV-2 infection, broadly referred to as Long COVID (LC). Although the frequencies of subsets of SARS-CoV-2-specific T cells have been shown to differ in individuals with LC relative to those with complete recovery, a deep dive into phenotypic and functional features of total and SARSCoV- 2-specific T cells from individuals with LC has yet to be performed. Method(s): Here, we used CyTOF to characterize the phenotypes and effector functions of T cells from LIINC cohort. The median age was 46, the cohort was 55.8% female, and 9/43 had been hospitalized. Participants were reported a median of 7 LC symptoms at 8 months. SARS-CoV-2-specific total antibody levels were also measured in concurrent sera. Manual gating was used to define T cell subsets, SPICE analyses for polyfunctionality, T cell clustering for phenotypic features, and linear regression for correlation. Permutation tests, Student's t tests, and Welch's t test were used for statistical analysis. Result(s): SARS-CoV-2 total antibody responses were elevated in the LC group (p=0.043), and correlated with frequencies of SARS-CoV-2-specific T cells in those without LC (r=0.776, p< 0.001) but not those with LC. While the frequencies of total SARS-CoV-2-specific CD4+ and CD8+ T cells were similar between individuals with and without LC, those from individuals without LC tended to be more polyfunctional (co-expressing IFNgamma, TNFalpha, IL2, and/or MIP1beta). CD4+ T cells from individuals with LC harbored higher frequencies of Tcm (p=0.003), Tfh (p=0.037), and Treg subsets (p=0.0412), and preferentially expressed a variety of tissue homing receptors including CXCR4 and CXCR5 (p=0.037). SARS-CoV-2-specific CD4+ T cells producing IL6, albeit rare, were observed exclusively among those with LC (p=0.016). In addition, participants with LC harbored significantly higher frequencies of SARS-CoV-2-specific CD8+ T cells co-expressing exhaustion markers PD1 and CTLA4 (p=0.018). Conclusion(s): Long COVID is characterized by global phenotypic differences in the CD4+ T cell compartment in ways suggesting preferential migration of these cells to inflamed mucosal tissues. Individuals with LC also harbor higher numbers of exhausted SARS-CoV-2-specific CD8+ T cells, potentially implicating viral persistence. Finally, our data additionally suggest that individuals with LC may uniquely exhibit an uncoordinated T cell and antibody response during COVID-19 convalescence.

DELINEATING FACTORS ASSOCIATED WITH ASYMPTOMATIC PRIMARY SARS-CoV-2 INFECTION

Background: SARS-CoV-2 infection leads to a broad range of clinical manifestations, from no symptoms to critical illness. Pre-stimulated innate defenses, rapid induction of SARS-CoV-2 responses and pre-existing crossreactive immunity to circulating human coronaviruses (HCoV) may early dampen SARS-CoV-2 infection, preventing symptomatic disease. Here we explore SARS-CoV-2 and HCoV antibody impact on asymptomatic infection in individuals first encountering SARS-CoV-2 in March 2020-March 2021 participating in a longitudinal pediatric cohort (n=2917) and a cross-sectional adult and children diagnostic cohort (n=882) in Switzerland. Method(s): Antibodies (Ab) to S1 of the four HCoV strains and SARS-CoV-2 antigens S1, RBD, S2 and N were determined in saliva (n=4993) and serum (n=7486) samples. Mucosal and systemic neutralization activity against wildtype SARS-CoV-2-Wuhan, and Alpha, Delta and Omicron (BA.2) variants of concerns (VoC) was assessed by pseudovirus neutralization in a subset of individuals. Result(s): Analysis of simultaneously sampled saliva and plasma revealed a strong correlation of mucosal and systemic SARS-CoV-2 anti-spike responses in recent infection. Notably, pre-existing high HCoV antibody response was significantly associated with higher systemic (IgG, IgA, and IgM, p< 0.001) and mucosal (IgG, p=0.03) SARS-CoV-2 responses following SARS-CoV-2 infection. Adjusting for age and sex, we found four saliva SARS-CoV-2 Ab parameters, namely total Ig S2, total Ig S1, IgA S2 and IgM S1 (p< 0.001, p< 0.001, p=0.02, p=0.01 respectively), inversely associated with salivary viral load highlighting the impact of mucosal Ab response on viral suppression. Saliva neutralization activity was modest but surprisingly broad, retaining same level activity against Wuhan, Alpha and Delta, but not Omicron BA.2, whereas plasma neutralization activity showed the typical decrease for all three VoCs compared to Wuhan. Most interestingly, asymptomatic individuals presented with higher IgG S2 antibody reactivity in saliva (p=0.03) and higher pre-existing HCoV-S1 IgG activity in plasma (HKU1, p=0.04) and saliva (total hCoV, p=0.02) suggesting immune factors that restrict disease severity. Conclusion(s): Focusing on a SARS-CoV-2 infection and vaccine naive population, our study reveals interdependencies of systemic and mucosal SARS-CoV-2 and HCoV immunity following primary SARS-CoV-2 infection and locates reactivities linked with reduced viral load and asymptomatic infection.

Cardiovascular complications are the primary drivers of mortality in hospitalized patients with SARS-CoV-2 community-acquired pneumonia.

BACKGROUND: Hospitalized patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) community-acquired pneumonia (CAP) and associated comorbidities are at increased risk of cardiovascular complications. The magnitude of effect of cardiovascular complications and the role of prior comorbidities on clinical outcomes are not well defined. RESEARCH QUESTION: What is the impact of cardiovascular complications on mortality in hospitalized patients with SARS-CoV-2 CAP? What is the impact of co-morbidities and other risk factors on the risk of developing cardiovascular complications and mortality in these patients? STUDY DESIGN AND METHODS: This cohort study included 1,645 hospitalized patients with SARS-CoV-2 CAP. Cardiovascular complications were evaluated. The clinical course during hospitalization was described using a multistate model with 4 states: hospitalized with no cardiovascular complications, hospitalized with cardiovascular complications, discharged alive, and dead. Cox proportional hazards regression was used to analyze the impact of prior comorbid conditions on transitions between these states. Hazard ratios (HRs) and 95% confidence intervals (CIs) were reported. RESULTS: Cardiovascular complications occurred in 18% of patients hospitalized with SARS-CoV-2 CAP. The mortality rate in this group was 45% versus 13% in patients without cardiovascular complications. Males (HR: 1.32, 95% CI: 1.03-1.68), older adults (HR: 1.34, 95% CI: 1.03-1.75), patients with congestive heart failure (HR: 1.59, 95% CI: 1.18-2.15), coronary artery disease (HR: 1.34, 95% CI: 1.00-1.79), atrial fibrillation (HR: 1.43, 95% CI: 1.06-1.95), direct admissions to the ICU (HR: 1.77, 95% CI: 1.36-2.32) and PaO2/FiO2 less than 200 (HR: 1.46, 95% CI: 1.11-1.92) were more likely to develop cardiovascular complications after hospitalization for SARS-CoV-2 CAP; however, these factors are not associated with increased risk of death after a cardiovascular complication.

Virtual Game Jam: Collaborative Pathway to Serious Games for Health

Game jams are collaborative competitions where participants make games from scratch within a short period of time. Occasionally, these games aim to promote meaningful learning outcomes by embedding functional knowledge into entertaining and engaging gameplay. We wanted to understand if a game jam during a global pandemic could be used as an effective method to facilitate a collaborative, multidisciplinary team's ability to design, develop, and implement a serious game on a health-related topic (i.e., COVID-19). We examined the 2020 game jam hosted by the National Academy of Sciences to evaluate one method to organize a serious virtual game jam and provide a roadmap for implementation using qualitative and quantitative data collection methods to evaluate its efficacy. In addition to identifying best practices and lessons learned from our tools and processes, our results demonstrate that a virtual game jam can in fact be a powerful tool for creating a scientifically sound, population-level, big picture-thinking serious game used for health education, facilitate collaborative multidisciplinary teams and be a practical learning experience for game jammers.

Simulation of COVID-19 Ultraviolet Disinfection Using Coupled Ray Tracing and CFD

Ultraviolet Germicidal Irradiance (UVGI) is the effective technique of inactivating disease-causing bacteria, mould spores, fungi, and viruses using ultraviolet radiation. In this study, we seek to quantify the efficacy and COVID-19 infection risk reduction achieved by UVGI in the upper unoccupied zone of a room so that we may specify the type and placement of UVGI emitters optimally. We present a computational fluid dynamics (CFD) based approach to model disinfection of aerosolized pathogens in a non-uniform ultraviolet field with mixing driven by air exchange and temperature gradients. We validate our CFD against simple calculation methods for UVGI effectiveness in well mixed spaces, and we integrate it with the Wells-Riley model of airborne infection risk to assess the relative benefit of UVGI with and against other measures. We demonstrate an order of magnitude reduction in infection risk as a result of applying UVGI, as well as the ability to quantify infection risk in non-well-mixed settings where simplified calculations methods do not apply. © International Building Performance Simulation Association, 2022

Forum: Gendered Dynamics of Academic Networks

This forum examines whether scholars' access to networks in the international studies profession is gendered and if so, the consequences of those networks for personal and professional success. Academic networks that encompass both professional and personal connections have been proposed as one solution to chilly climate issues because they provide a dual function of enhancing scholarly productivity and inclusion in the profession. The articles in the forum consider both professional (e.g. citation) and personal (e.g. mentorship, friendship) networks, as well as traditional (e.g. invited talks) and nontraditional (e.g. social media) networks. The authors show that biases that arise through the gendered nature of academic networks can be mitigated through social media, mentoring, and friendship networks. However, we must also be cognizant of other factors that create barriers for women in the profession (e.g. university prestige, parenthood, COVID-19). [ FROM AUTHOR] Copyright of International Studies Perspectives is the property of Oxford University Press / USA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full . (Copyright applies to all s.)

Novel recombinant SARS-CoV-2 lineage detected through genomic surveillance in Wales, UK.

Recombination, the process whereby a segment of genetic material from one genome is inserted into another, producing a new chimeric genome, is an important evolutionary mechanism frequently observed in coronaviruses. The risks posed by recombination include the shuffling of advantageous mutations that may increase transmissibility, severity or vaccine escape. We present a genomic and epidemiological description of a new recombinant lineage of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), XR, first identified in Wales. The Pathogen Genomics Unit (Public Health Wales, UK) sequences positive SARS-CoV-2 PCR tests using the ARTIC SARS-CoV-2 sequencing protocol. Recombinants were detected using an in-house pipeline and the epidemiological data analysed in R. Nosocomial cases were defined as those with samples taken after >7 days in hospital. Between February and March 2022, we identified 78 samples with highly similar genomes, comprising a BA.1-like 5' end, a BA.2-like 3' end and a BA.2-like spike protein. This signature is consistent with recombination and was defined as XR by Pangolin (PANGO v1.8). A total of 50 % of cases had a sample collected whilst in hospital and the first three cases were immunocompromised patients. The patient median age was 58 years (range: 4-95 years) and most of the patients were fully vaccinated against SARS-CoV-2 (74 % third dose/booster). Three patients died within 28 days of their sample collection date, one of whom had COVID-19 listed amongst ICD10 (International Classification of Diseases 10) coded causes of death. Our integrated system enabled real-time monitoring of recombinant SARS-CoV-2 for early detection, in order to rapidly risk assess and respond. This work highlights the importance of setting-based surveillance of recombinant SARS-CoV-2, as well as the need to monitor immunocompromised populations through repeat testing and sequencing.

The road to achieving epidemic-ready primary health care.

Millions of avoidable deaths arising from the COVID-19 pandemic emphasise the need for epidemic-ready primary health care aligned with public health to identify and stop outbreaks, maintain essential services during disruptions, strengthen population resilience, and ensure health worker and patient safety. The improvement in health security from epidemic-ready primary health care is a strong argument for increased political support and can expand primary health-care capacities to improve detection, vaccination, treatment, and coordination with public health-needs that became more apparent during the pandemic. Progress towards epidemic-ready primary health care is likely to be stepwise and incremental, advancing when opportunity arises based on explicit agreement on a core set of services, improved use of external and national funds, and payment based in large part on empanelment and capitation to improve outcomes and accountability, supplemented with funding for core staffing and infrastructure and well designed incentives for health improvement. Health-care worker and broader civil society advocacy, political consensus, and bolstering government legitimacy could promote strong primary health care. Epidemic-ready primary health-care infrastructure that is able to help prevent and withstand the next pandemic will require substantial financial and structural reforms and sustained political and financial commitment. Governments, advocates, and bilateral and multilateral agencies should seize this window of opportunity before it closes.

Aspergillus fumigatus Supernatants Disrupt Bronchial Epithelial Monolayers: Potential Role for Enhanced Invasion in Cystic Fibrosis.

Aspergillus fumigatus is the most commonly isolated fungus in chronic lung diseases, with a prevalence of up to 60% in cystic fibrosis patients. Despite this, the impact of A. fumigatus colonisation on lung epithelia has not been thoroughly explored. We investigated the influence of A. fumigatus supernatants and the secondary metabolite, gliotoxin, on human bronchial epithelial cells (HBE) and CF bronchial epithelial (CFBE) cells. CFBE (F508del CFBE41o-) and HBE (16HBE14o-) trans-epithelial electrical resistance (TEER) was measured following exposure to A. fumigatus reference and clinical isolates, a gliotoxin-deficient mutant (ΔgliG) and pure gliotoxin. The impact on tight junction (TJ) proteins, zonula occludens-1 (ZO-1) and junctional adhesion molecule-A (JAM-A) were determined by western blot analysis and confocal microscopy. A. fumigatus conidia and supernatants caused significant disruption to CFBE and HBE TJs within 24 h. Supernatants from later cultures (72 h) caused the greatest disruption while ΔgliG mutant supernatants caused no disruption to TJ integrity. The ZO-1 and JAM-A distribution in epithelial monolayers were altered by A. fumigatus supernatants but not by ΔgliG supernatants, suggesting that gliotoxin is involved in this process. The fact that ΔgliG conidia were still capable of disrupting epithelial monolayers indicates that direct cell-cell contact also plays a role, independently of gliotoxin production. Gliotoxin is capable of disrupting TJ integrity which has the potential to contribute to airway damage, and enhance microbial invasion and sensitisation in CF.

Low-density lipoprotein receptor-related protein 1 (LRP1) as an auxiliary host factor for RNA viruses.

Viruses with an RNA genome are often the cause of zoonotic infections. In order to identify novel pro-viral host cell factors, we screened a haploid insertion-mutagenized mouse embryonic cell library for clones that are resistant to Rift Valley fever virus (RVFV). This screen returned the low-density lipoprotein receptor-related protein 1 (LRP1) as a top hit, a plasma membrane protein involved in a wide variety of cell activities. Inactivation of LRP1 in human cells reduced RVFV RNA levels already at the attachment and entry stages of infection. Moreover, the role of LRP1 in promoting RVFV infection was dependent on physiological levels of cholesterol and on endocytosis. In the human cell line HuH-7, LRP1 also promoted early infection stages of sandfly fever Sicilian virus and La Crosse virus, but had a minor effect on late infection by vesicular stomatitis virus, whereas encephalomyocarditis virus was entirely LRP1-independent. Moreover, siRNA experiments in human Calu-3 cells demonstrated that also SARS-CoV-2 infection benefitted from LRP1. Thus, we identified LRP1 as a host factor that supports infection by a spectrum of RNA viruses.