Optimisations and challenges involved in the creation of various bioluminescent and fluorescent influenza a virus strains for in vitro and in vivo applications

Bioluminescent and fluorescent influenza A viruses offer new opportunities to study influenza virus replication, tropism and pathogenesis. To date, several influenza A reporter viruses have been described. These strategies typically focused on a single reporter gene (either bioluminescent or fluorescent) in a single virus backbone. However, whilst bioluminescence is suited to in vivo imaging, fluorescent viruses are more appropriate for microscopy. Therefore, the idea l reporter virus varies depending on the experiment in question, and it is important that any reporter virus strategy can be adapted accordingly. Herein, a strategy was developed to create five different reporter viruses in a single virus backbone. Specifically, enhanced green fluorescent protein (eGFP), far-red fluorescent protein (fRFP), near-infrared fluorescent protein (iRFP), Gaussia luciferase (gLUC) and firefly luciferase (fLUC) were inserted into the PA gene segment of A/PR/8/34 (H1N1). This study provides a comprehensive characterisation of the effects of different reporter genes on influenza virus replication and reporter activity. In vivo reporter gene expression, in lung tissues, was only detected for eGFP, fRFP and gLUC expressing viruses. In vitro, the eGFP-expressing virus displayed the best reporter stability and could be used for correlative light electron microscopy (CLEM). This strategy was then used to create eGFP-expressing viruses consisting entirely of pandemic H1N1, highly pathogenic avian influenza (HPAI) H5N1 and H7N9. The HPAI H5N1 eGFP-expressing virus infected mice and reporter gene expression was detected, in lung tissues, in vivo. Thus, this study provides new tools and insights for the creation of bioluminescent and fluorescent influenza A reporter viruses. Copyright

The psychosocial adaptability of independently living older adults to COVID-19 related social isolation in the Netherlands: A qualitative study

Since coronavirus disease 2019 (COVID-19) entered the Netherlands, the older adults (aged 70 or above) were recommended to isolate themselves, resulting in less social contact and possibly increased loneliness. The aim of this qualitative study was to explore independently living older adults' perceptions of social and emotional well-being during the COVID-19-related self-isolation, and their motivation to expand their social network in the future. Semi-structured phone interviews were held with 20 community-dwelling adults (age range 56-87; 55% female) between April and June 2020 in the Netherlands. The interviews were audio recorded and transcribed verbatim. Open coding process was applied to identify categories and themes. Participants said to use more digital technologies to maintain contacts and adapt to the government measurements. Most participants missed the lack of social contacts, while some participants had no problems with the reduced social contacts. The emotional well-being of most participants did not change. Some participants felt unpleasant or mentioned that the mood of other people had changed. Participants were not motivated to expand their social network because of existing strong networks. The relatively vital community-dwelling older adults in this study were able to adapt to the government recommendations for self-isolation with limited negative impact on their socio-emotional well-being

Older Adults' Views on Social Interactions and Online Socializing Games - A Qualitative Study.

Age-related difficulties and quarantine restrictions impede the possibilities to maintain contact with one's social network. Maintaining these contacts may be supported by digital games. To develop effective and feasible digital tools to foster social interaction, we aimed to explore what older adults find important in social contact and what barriers and enablers they foresee in digital gaming interventions as network support aids. Two focus groups and 20 semi-structured interviews (N = 29) with older adults (aged 55-87) were held to explore the research questions. Furthermore, a questionnaire was administered (N = 29) containing measures of loneliness, frailty, and social network size. Participants found 'reciprocity', 'in-person contact', and 'personal connection' important in contact with strong ties. Online games were not used much for socializing but may be used in the future, particularly by less mobile older adults. Future social gaming interventions should be challenging, user-friendly, and offer the possibility to communicate. Digital co-designed interventions that are feasible, challenging, intuitive, and trigger meaningful communication may strengthen social interactions in older adults. They may be a relevant social support tool in periods of interaction limitations due to functional impairment or social isolation

Impact of malaria control interventions on malaria infection and anaemia in low malaria transmission settings: a cross-sectional population-based study in Sudan

Background: The past two decades were associated with innovation and strengthening of malaria control interventions, which have been increasingly adopted at large scale. Impact evaluations of these interventions were mostly performed in moderate or high malaria transmission areas. This study aimed to evaluate the use and performance of malaria interventions in low transmission areas on malaria infections and anaemia. Methods: Data from the 2016 Sudan malaria indicator survey was used. Multi-level logistic regression analysis was used to assess the strength of association between real-life community-level utilization of malaria interventions [diagnosis, artemisinin-based combination therapies (ACTs) and long-lasting insecticidal nets (LLINs)] and the study outcomes: malaria infections and anaemia (both overall and moderate-to-severe anaemia). Results: The study analysis involved 26,469 individuals over 242 clusters. Malaria infection rate was 7.6%, overall anaemia prevalence was 47.5% and moderate-to-severe anaemia prevalence was 4.5%. The average community-level utilization was 31.5% for malaria diagnosis, 29.9% for ACTs and 35.7% for LLINs. The odds of malaria infection was significantly reduced by 14% for each 10% increase in the utilization of malaria diagnosis (adjusted odds ratio (aOR) per 10% utilization 0.86, 95% CI 0.78-0.95, p = 0.004). However, the odds of infection was positively associated with the utilization of LLINs at community-level (aOR per 10% utilization 1.20, 95% CI 1.11-1.29, p < 0.001). No association between malaria infection and utilization of ACTs was identified (aOR per 10% utilization 0.97, 95% CI 0.91-1.04, p = 0.413). None of the interventions was associated with overall anaemia nor moderate-to-severe anaemia. Conclusion: There was strong evidence that utilization of malaria diagnosis at the community level was highly protective against malaria infection. No protective effect was seen for community utilization of ACTs or LLINs. No association was established between any of the interventions and overall anaemia or moderate-to-severe anaemia. This lack of effectiveness could be due to the low utilization of interventions or the low level of malaria transmission in the study area. Identification and response to barriers of access and low utilization of malaria interventions are crucial. It is crucial to ensure that every suspected malaria case is tested in a timely way, notably in low transmission settings

Thrombo-Inflammation in Cardiovascular Disease: An Expert Consensus Document from the Third Maastricht Consensus Conference on Thrombosis

Thrombo-inflammation describes the complex interplay between blood coagulation and inflammation that plays a critical role in cardiovascular diseases. The third Maastricht Consensus Conference on Thrombosis assembled basic, translational, and clinical scientists to discuss the origin and potential consequences of thrombo-inflammation in the etiology, diagnostics, and management of patients with cardiovascular disease, including myocardial infarction, stroke, and peripheral artery disease. This article presents a state-of-the-art reflection of expert opinions and consensus recommendations regarding the following topics: (1) challenges of the endothelial cell barrier; (2) circulating cells and thrombo-inflammation, focused on platelets, neutrophils, and neutrophil extracellular traps; (3) procoagulant mechanisms; (4) arterial vascular changes in atherogenesis; attenuating atherosclerosis and ischemia/reperfusion injury; (5) management of patients with arterial vascular disease; and (6) pathogenesis of venous thrombosis and late consequences of venous thromboembolism.Thrombosis and Hemostasi