Investigating the function of neutrophils in RSV Bronchiolitis using a novel in vitro translational model of transepithelial immune cell migration

Background: Respiratory Syncytial Virus (RSV) infects almost all children before the age of 2. In most children, RSV infection causes a mild to moderate illness, but a small percentage of children develop severe RSV bronchiolitis and require hospitalisation. A significant proportion of these acutely unwell children were previously otherwise healthy. Clinical studies of infants with severe bronchiolitis have shown a massive infiltrate of neutrophils to the airway, comprising over 80% of all cells recovered. In this thesis I explore further whether the effect of neutrophil infiltration is necessary for viral clearance or detrimental to clinical disease, by exploring the interaction between neutrophils and RSV infected airway epithelial cells (AECs). Aims: The aims of this thesis were to (i) describe neutrophil movement across RSV infected AECs, (ii) measure the effect of interaction with RSV infected AECs on neutrophil apoptosis, (iii) activation, then (iv) compare these effects between adult and cord blood neutrophils. Methods: To examine the complex relationship between the RSV infected airway and neutrophil function, this study optimises a novel in-vitro transepithelial migration model of RSV infected AECs, comprising human neutrophils, ciliated epithelial cells cultured at air-liquid interface and RSV. This has allowed real-time video microscopy of neutrophil transepithelial migration, examination of dynamic neutrophil activation and assessment of neutrophil viability and apoptosis during infection. Results and Conclusion: Three populations of neutrophils interacting with AECs were described, those basolateral to AECs, those apical and those adhered to AECs. Imaging neutrophil migration revealed adhered neutrophils appear to kill virally infected AECs during migration. Neutrophil apoptosis and activation were defined by their interaction with AECs and evidence presented in this thesis suggest physiological differences between adult and cord blood neutrophil response to RSV infected AECs may predispose to a more severe disease picture in neonates

Sequence of the mouse Q4 class I gene and characterization of the gene product

The Q4 class I gene has been shown to participate in gene conversion events within the mouse major histocompatibility complex. Its complete genomic nucleotide sequence has been determined. The 5' half of Q4 resembles H-2 genes more strongly than other Q genes. Its 3' end, in contrast, is Q-like and contains a translational stop signal in exon 5 which predicts a polypeptide with an incomplete membrane spanning segment. The presence of two inverted B1 repeats suggests that part of the Q4 gene may be mobile within the genome. Gene transfer experiments have shown that the Q4 gene encodes a ß2-microglobulin associated polypeptide of Mr 41 000. A similar protein was found in activated mouse spleen cells. The Q4 polypeptide was found to be secreted both by spleen cells and by transfected fibroblasts and was not detectable on the cell surface. Antibody binding and twodimensional gel electrophoresis indicate that the Q4 molecule is identical to a mouse class I polypeptide, Qb-1, which has been previously described

Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24.

Off-target binding of hydrophobic drugs can lead to unwanted side effects, either through specific or non-specific binding to unintended membrane protein targets. However, distinguishing the binding of drugs to membrane proteins from that of detergents, lipids and cofactors is challenging. Here, we use high-resolution mass spectrometry to study the effects of HIV protease inhibitors on the human zinc metalloprotease ZMPSTE24. This intramembrane protease plays a major role in converting prelamin A to mature lamin A. We monitored the proteolysis of farnesylated prelamin A peptide by ZMPSTE24 and unexpectedly found retention of the C-terminal peptide product with the enzyme. We also resolved binding of zinc, lipids and HIV protease inhibitors and showed that drug binding blocked prelamin A peptide cleavage and conferred stability to ZMPSTE24. Our results not only have relevance for the progeria-like side effects of certain HIV protease inhibitor drugs, but also highlight new approaches for documenting off-target drug binding

The embedding of universities in innovation ecosystems: The case of marine research at the University of Bergen

While historically the core missions of universities have been research and teaching, it has become increasingly recognised that universities have become significant sources of knowledge and capabilities. This third mission is cementing the role of universities as suppliers of qualified labour and generators of knowledge and technologies that promote innovation in a variety of innovation ecosystems. The main goal of the paper is to illustrate an approach that captures the various contributions of universities to their innovation ecosystems. Often territorially bounded, such links provide insights into the characteristics and geography of the various linkage for a university. With the case of the University of Bergen and its role within the marine innovation ecosystem of Western Norway, this ‘ecosystem fingerprint’, can be seen as a useful means to clarify the third mission of universities through the linkages and interdependencies with various actors. The authors demonstrate that a university can act both as a global pipeline provider and take active part in the local buzz, providing this concept with new empirical insight. The authors conclude that the university is highly embedded in both the marine innovation ecosystem and the knowledge ecosystem, but with linkages extended to interconnected business ecosystems.publishedVersio

Medieval Convent Drama: Translating Scripture and Transforming the Liturgy

This article examines a vernacular Nativity play from the convent at Huy, in modern- day Belgium. The play includes liturgical citations that would have been sung in Latin, alongside the Walloon French lines that translate generically -- scriptural narrative into verse drama -- as well as linguistically. Scriptural narrative is also expanded through the addition of apocryphal material and especially female characters. The Huy play survives in a manuscript of the 15th century, and in a seventeenth-century manuscript that reveals further processes of revision and adaptation by the nuns who composed, copied and performed it. This paper describes these various processes of translation and adaptation for performance by and for female religious, and comments on theatrical translation and the relationship of drama to liturgy

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Converging evidence has accumulated that menstrual cycle and thus hormonal levels can affect emotional behavior, in particular facial emotion recognition. Here we explored the association of ovarian hormone levels and amygdala activation during an explicit emotion recognition task in two groups of healthy young females: one group was measured while in their follicular phase (n = 11) and the other during their luteal phase (n = 11). Using a 3T scanner in combination with a protocol specifically optimized to reliably detect amygdala activation we found significantly stronger amygdala activation in females during their follicular phase. Also, emotion recognition performance was significantly better in the follicular phase. We observed significant negative correlations between progesterone levels and amygdala response to fearful, sad and neutral faces, further supporting a significant modulation of behavior and neural response by hormonal changes during the menstrual cycle. From an evolutionary point of view this significant influence of ovarian hormone level on emotion processing and an important neural correlate, the amygdala, may enable a higher social sensitivity in females during their follicular phase, thus facilitating socio-emotional behavior (and social interaction) which may possibly facilitate mating behavior as well

Tumour growth in mice resistant to diet-induced obesity

Obesity is a chronic disease with associated increases in the incidence, and a reduction in survival, of many cancer types. Obesity results from an imbalance in calorie intake and calorie requirement. This study aimed to investigate the separate effects of high-fat diet and obesity on cancer in an animal model resistant to diet-induced obesity. Male BALB/c mice fed long-term on a high-fat, Western-style diet were implanted with syngeneic CT26 colon adenocarcinoma cells and compared to mice fed normal diet. BALB/c mice on high-fat diet were 10% heavier than mice fed normal diet, with no difference in tumour growth rates or tumour cell proliferation. Subgroups of mice that became obese on high-fat diet, however, showed increased tumour growth rates compared to mice fed normal diet, whereas mice that remained slim showed no difference in tumour growth. Protein arrays identified several adipokines that were expressed at different levels, including serum Tissue Inhibitors of Metallo-Proteinases (TIMP-1) and tumour C-Reactive Protein (CRP). In conclusion, tumour growth was enhanced in mice unable to resist obesity, and adipokine profiles were affected by the animals’ ability to resist obesity

Rural resilience in a digital society: Editorial

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Dr. Conway et al reply

We thank Sauret et al for their interest in our systematic literature review that explored potential diagnostic confusion between giant cell arteritis (GCA) and the coronavirus disease 2019 (COVID-19). This was a particularly important consideration during the early months of the COVID-19 pandemic, when community testing for SARS-CoV-2 was limited and diagnostic tests for GCA were restricted or unavailable due to redeployment of staff.</p