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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) encodes six accessory proteins (3a, 6, 7a, 7b, 8, and 9b) for which limited information is available on their role in pathogenesis. We showed that the deletion of open reading frames (ORFs) 6, 7a, or 7b individually did not significantly impact viral pathogenicity in humanized K18-hACE2 transgenic mice. In contrast, the deletion of ORF8 partially attenuated SARS-CoV-2, resulting in reduced lung pathology and 40% less mortality, indicating that ORF8 is a critical determinant of SARS-CoV-2 pathogenesis. Attenuation of SARS-CoV-2-∆8 was not associated with a significant decrease in replication either in the lungs of mice or in organoid-derived human airway cells. An increase in the interferon signaling at early times post-infection (1 dpi) in the lungs of mice and a decrease in the pro-inflammatory and interferon response at late times post-infection, both in the lungs of mice (6 dpi) and in organoid-derived human airway cells [72 hours post-infection (hpi)], were observed. The early, but not prolonged, interferon response along with the lower inflammatory response could explain the partial attenuation of SARS-CoV-∆8. The presence of ORF8 in SARS-CoV-2 was associated with an increase in the number of macrophages in the lungs of mice. In addition, the supernatant of SARS-CoV-2-WT (wild-type)-infected organoid-derived cells enhanced the activation of macrophages as compared to SARS-CoV-2-∆8-infected cells. These results show that ORF8 is a virulence factor involved in inflammation that could be targeted in COVID-19 therapies. IMPORTANCE The relevance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ORF8 in the pathogenesis of COVID-19 is unclear. Virus natural isolates with deletions in ORF8 were associated with wild milder disease, suggesting that ORF8 might contribute to SARS-CoV-2 virulence. This manuscript shows that ORF8 is involved in inflammation and in the activation of macrophages in

The production of type I interferon (IFN) is the hallmark of the innate immune response. Most, if not all, mammalian viruses have a way to circumvent this response. Fundamental knowledge on viral evasion of innate immune responses may facilitate the design of novel antiviral therapies. To investigate how human metapneumovirus (HMPV) interacts with the innate immune response, recombinant viruses lacking G, short hydrophobic (SH), or M2-2 protein expression were assessed for IFN induction in A549 cells. HMPV lacking G or SH protein expression induced similarly low levels of IFN, compared to the wild-type virus, whereas HMPV lacking M2-2 expression induced significantly more IFN than the wild-type virus. However, sequence analysis of the genomes of M2-2 mutant viruses revealed large numbers of mutations throughout the genome. Over 70% of these nucleotide substitutions were A-to-G and T-to-C mutations, consistent with the properties of the adenosine deaminase acting on RNA (ADAR) protein family. Knockdown of ADAR1 by CRISPR interference confirmed the role of ADAR1 in the editing of M2-2 deletion mutant virus genomes. More importantly, Northern blot analyses revealed the presence of defective interfering RNAs (DIs) in M2-2 mutant viruses and not in the wild-type virus or G and SH deletion mutant viruses. DIs are known to be potent inducers of the IFN response. The presence of DIs in M2-2 mutant virus stocks and hypermutated virus genomes interfere with studies on HMPV and the innate immune response and should be addressed in future studies. IMPORTANCE Understanding the interaction between viruses and the innate immune response is one of the barriers to the design of antiviral therapies. Here, we investigated the role of the G, SH, and M2-2 proteins of HMPV as type I IFN antagonists. In contrast to other studies, no IFN-antagonistic functions could be observed for the G and SH proteins. HMPV with a deletion of the M2-2 protein did induce type I IFN production upon infe

Patagonia, is an imaginary territory that comprises the south of Chile and Argentina. Historically, Patagonia has captivated an image of far-off lands and wilderness, embedded in a remote and peripheral context, which in turn has often been idealized as ‘pristine’. In the literature, the reader will find different terminologies referring to Patagonia, in both Chile and Argentina and diverse ideas of where the imaginary boundaries of Patagonia reside. In this dissertation I focus on the Chilean territory, specifically the territory that comprises the administrative region of Aysén, also known as Northern Patagonia. Hereafter, Aysén-Patagonia.The current social-ecological state of Aysén-Patagonia, cannot be fully understood without looking into historical processes of idealization, territorial (re)invention and discursive transformations, which have molded the common perception towards the Aysén region, and at the same time have influenced how the region (self)transforms.Over the years, Aysén-Patagonia has faced a series of socio-spatial transformations driven by development and environmental ideas. From a development point of view, Chilean Patagonia, and specifically Northern Patagonia transitioned from extensive livestock ranching in the early twentieth century, to small-scale agriculture, nature-based tourism and industrial salmon aquaculture. From an environmental perspective, cultural and natural heritage associated to the rugged terrains and the livestock farming traditions are being replaced with renovated meanings of pristine and wild nature, situated within national and global targets of nature conservation.Despite the particular socio-spatial transformations that have taken place in Aysén-Patagonia, no studies have explored the entanglement of environmental and development ideas alike, nor their effects on the Aysén’s peripheral condition. This research contributes to fill this gap, by delving into three current prominent aspects for the environment and deve

Werkstress wordt vaak verward met werkdruk. Een te hoge werkdruk kan leiden tot werkstress, maar dit hoeft zeker niet altijd het geval te zijn. Werkdruk en stress zijn sterk individueel bepaald; geen twee werknemers hebben in gelijke mate last van stress. Hier beschrijven we hoe je het herkent en nog belangrijker wat je kunt doen om dit te voorkomen.

Human respiratory syncytial virus (HRSV) is the leading cause of bronchioli- tis in infants. Two subgroups of HRSV (A and B) routinely cocirculate. Most research has been performed with HRSV-A strains because these are easier to culture than HRSV-B strains. In this study, we aimed to compare the replicative fitness and HRSV-induced innate cytokine responses of HRSV-A and HRSV-B strains in disease-relevant cell culture models. We used two recombinant (r) clinical isolate-based HRSV strains (A11 and B05) and one recombinant laboratory-adapted HRSV strain (A2) to infect commercially avail- able nasal, bronchial, and small-airway cultures. Epithelial cells from all anatomical loca- tions were susceptible to HRSV infection despite the induction of a dominant type III interferon response. Subgroup A viruses disseminated and replicated faster than the subgroup B virus. Additionally, we studied HRSV infection and innate responses in air- way organoids (AOs) cultured at air-liquid interface (ALI). Results were similar to the commercially obtained bronchial cells. In summary, we show that HRSV replicates well in cells from both the upper and the lower airways, with a slight replicative advantage for subgroup A viruses. Lastly, we showed that AOs cultured at ALI are a valuable model for studying HRSV ex vivo and that they can be used in the future to study factors that influence HRSV disease severity. IMPORTANCE Human respiratory syncytial virus (HRSV) is the major cause of bron- chiolitis and pneumonia in young infants and causes almost 200,000 deaths per year. Currently, there is no vaccine or treatment available, only a prophylactic mono- clonal antibody (palivizumab). An important question in HRSV pathogenesis research is why only a fraction (1 to 3%) of infants develop severe disease. Model systems comprising disease-relevant HRSV isolates and accurate and reproducible cell culture models are indispensable to study infection, replication, and innate immune responses. H

Against the background of overall decline and inadequacy of funding for nature conservation, the budgets of the five largest nature conservation organisations have been growing significantly since the 1990s to command over 50% of globally available conservation funding. This thesis focusses on World Wide Fund for Nature (WWF) to show how large nature conservation organisations maintain and expand funding for global biodiversity conservation. It begins exploratorily with a thematic review of 64 peer-reviewed articles to find dominant themes recurring around the funding of biodiversity conservation between 2010-2016. The principle finding is that the main challenge in biodiversity financing is not only underfunding but that quite often available funds do not go where they are most needed. Conservation networks play a key role in determining financial flows. This informs the choice for a conceptual framework that is inspired by the network theory postulated by Manuel Castells. This thesis examines how WWF uses networking to maintain and expand funding from public sources, and then shows how much financial income WWF receives from the corporate sector in comparison to other income streams, including from market-based approaches. The main finding is that WWF has been successful in increasing its funding from the public sector at 7.5% p.a. by being in “right networks”, speaking the “right language”, and connecting to “relevant flows”, including dominant governmental ministries, bilateral and multilateral organisations, emerging and graduated economies and the private sector that relates closely with them. However, WWF’s income from the corporate sector has remained low at 11% p.a. and stagnant, and its “bankable projects” are yet to produce any income and remain philanthropic. WWF’s most important funding base is private small donors who contribute 40% of its income. Based on these findings, this thesis points to a possible scenario on the future of conservation finance t

Dichte grenzen, gecancelde vakanties en de onzekerheid rondom de coronamaatregelen hebben ervoor gezorgd dat veel werknemers het afgelopen jaar ervoor hebben gekozen om door te werken en genoegen te nemen met korte vakanties. Op zich een positieve zaak, zou je zeggen, zeker als er werk genoeg is, maar wat als de verlofsaldi maar blijven groeien?

Als werkgever hoop je het nooit mee te maken: een werknemer met wie er problemen zijn in de werksfeer. Soms zit er zelfs niets anders op dan een werknemer de deur te wijzen. Hoe kan het waarschuwingsgesprek je helpen en grotere problemen voorkomen?

Item does not contain fulltext, BACKGROUND: Variants in the Structural Maintenance of Chromosomes flexible Hinge Domain-containing protein 1 (SMCHD1) can cause facioscapulohumeral muscular dystrophy type 2 (FSHD2) and the unrelated Bosma arhinia microphthalmia syndrome (BAMS). In FSHD2, pathogenic variants are found anywhere in SMCHD1 while in BAMS, pathogenic variants are restricted to the extended ATPase domain. Irrespective of the phenotypic outcome, both FSHD2-associated and BAMS-associated SMCHD1 variants result in quantifiable local DNA hypomethylation. We compared FSHD2, BAMS and non-pathogenic SMCHD1 variants to derive genotype-phenotype relationships. METHODS: Examination of SMCHD1 variants and methylation of the SMCHD1-sensitive FSHD locus DUX4 in 187 FSHD2 families, 41 patients with BAMS and in control individuals. Analysis of variants in a three-dimensional model of the ATPase domain of SMCHD1. RESULTS: DUX4 methylation analysis is essential to establish pathogenicity of SMCHD1 variants. Although the FSHD2 mutation spectrum includes all types of variants covering the entire SMCHD1 locus, missense variants are significantly enriched in the extended ATPase domain. Identification of recurrent variants suggests disease-specific residues for FSHD2 and in BAMS, consistent with a largely disease-specific localisation of variants in SMCHD1. CONCLUSIONS: The localisation of missense variants within the ATPase domain of SMCHD1 may contribute to the differences in phenotypic outcome.

Misleading claims about mass migration induced by climate change continue to surface in both academia and policy. This requires a new research agenda on ‘climate mobilities’ that moves beyond simplistic assumptions and more accurately advances knowledge of the nexus between human mobility and climate change.

Environmental conditions in Polar Regions are becoming more dynamic due to climate change. As sea ice melts, the range of human activities in Polar Regions are projected to increase, while weather conditions are becoming more extreme and unpredictable. Provision and use of weather, water, ice and climate (WWIC) information plays a key role in ensuring that polar activities are conducted as safely as possible and can contribute to a reduction of the environmental footprint of human activities. In this article, we explore the WWIC information provider landscape in a polar context, drawing on a database we compiled to characterize the diversity of providers. The database is built on available literature and on an extensive desk-based research of WWIC information provider websites. We analyse the 374 providers categorized by (a) institutional background (public vs private), (b) the position of the provider relative to activities in the WWIC information space, and (c) the users they serve. While governmental institutions have a strong presence in information provision, new types of providers are now entering the scene. Scientific actors seem to play a substantial role as users as well as major providers of WWIC information services.

Phospholipase C (PLC) has been suggested to play important roles in plant stress and development. To increase our understanding of PLC signaling in plants, we have started to analyze knock-out (KO), knock-down (KD) and overexpression mutants of Arabidopsis thaliana, which contains nine PLCs. Earlier, we characterized PLC2, PLC3 and PLC5. Here, the role of PLC7 is functionally addressed. Promoter-GUS analyses revealed that PLC7 is specifically expressed in the phloem of roots, leaves and flowers, and is also present in trichomes and hydathodes. Two T-DNA insertion mutants were obtained, i.e., plc7-3 being a KO- and plc7-4 a KD line. In contrast to earlier characterized phloem-expressed PLC mutants, i.e., plc3 and plc5, no defects in primary- or lateral root development were found for plc7 mutants. Like plc3 mutants, they were less sensitive to ABA during stomatal closure. Double-knockout plc3 plc7 lines were lethal, but plc5 plc7 (plc5/7) double mutants were viable, and revealed several new phenotypes, not observed earlier in the single mutants. These include a defect in seed mucilage, enhanced leaf serration, and an increased tolerance to drought. Overexpression of PLC7 enhanced drought tolerance too, similar to what was earlier found for PLC3-and PLC5 overexpression. In vivo 32Pi-labeling of seedlings and treatment with sorbitol to mimic drought stress, revealed stronger PIP2 responses in both drought-tolerant plc5/7 and PLC7-OE mutants. Together, these results show novel functions for PLC in plant stress and development. Potential molecular mechanisms are discussed.