Vaccinia virus protein B14, an inhibitor of NF-κB activation, and its counterpart in MVA

Vaccinia virus (VACV) encodes multiple proteins which modulate NF-[kappa]B activation to evade the host immune response. One example is B14, a virulence factor that reduces NF-[kappa]B activation via interaction with IKK [Beta]. B14 has orthologues in many VACV strains, including modified virus Ankara (MVA), which lacks many of the immunomodulators present in other VACV strains. Here, the MVA counterpart of B14, protein 183, was characterised. 183R was both removed from MVA and inserted into the B14R locus of a VACV strain Western Reserve (WR) lacking B14R, but in each case there was no change of phenotype. Protein 183 shares 95 % amino acid identity with B14 but, unlike B14, was unstable in eukaryotic cells, unless protein degradation was inhibited. Protein 183 did not inhibit NF-[kappa] B activation in response to cytokine stimulation, as does B14, nor did it restore the virulence phenotype of WR lacking B14R back to wild type. Therefore, the mutations incurred by 183 during the derivation of MVA have rendered the protein non-functional. However, other MVA immunomodulators remain to be characterised and this thesis describes a bacterial artificial chromosome (BAC) system that may facilitate the improvement of MVA in its role as a vaccine vector. To further characterise VACV modulation of NF-[kappa] B, a VACV WR strain was constructed lacking B14 and another VACV modulator of NF-[kappa]B, WR A49. Initial characterisation showed no change with this mutant virus in replication or spread. The effect of the B14 on NF-[kappa]B was further characterised by studying the B14-IKK [Beta] interaction using IKK[Beta] and B14 mutants. Residues 300 to 480 of IKK [Beta] were shown to be required for the interaction and a mutant encompassing this region co-purified to a degree with B14 when expressed in E. coli. In addition, mutagenesis showed that B14 residue F130 is required for the interaction of the two proteins

Painting with words: portraits of adult singers' perceptions on meaningfulness in two community choruses

The purpose of this study was to investigate what adult singers find meaningful about their participation in a volunteer community chorus and to consider the implication of the choral place on participant sense of meaningfulness. The researcher collected data from four adult singers, their family members, and two conductors. Bridging the gap between science and art, the researcher used a qualitative research method called portraiture, wherein narrative portraits are developed and written by a researcher in a descriptive manner providing voice to understanding the phenomenon of interest, to create portraits of four adult singers. Portraiture researchers seek to discover resonant stories within the complicated framework of daily life. Semi-structured interviews with each participant provided data. In addition, adult singers kept personal journals for the researcher, and the researcher observed participant behaviors before, during, and after rehearsals. The researcher implemented axial, focused, and theoretical analysis during data analysis and interpretation to construct a narrative portrait of each participating singer reflecting the emergent patterns and themes. The two emergent themes were connectedness and achievement. Overall, connections with other people and achieving surmountable challenges were highly valued aspects of this study. Further analysis for the impact of sense of place on meaningfulness noted the impact of the choral place and the choral process on participants’ perceptions of meaningfulness

Vaccine responses in ageing and chronic viral infection

Over the last few decades, changing population demographics have shown that there are a growing number of individuals living past the age of 60. With this expanding older population comes an increase in individuals that are more susceptible to chronic illness and disease. An important part of maintaining health in this population is through prophylactic vaccination, however, there is growing evidence that vaccines may be less effective in the elderly. Furthermore, with the success of anti-viral therapies, chronic infections such as HIV are becoming increasingly prevalent in older populations and present a relatively unstudied population with respect to the efficacy of vaccination. Here we will examine the evidence for age-associated reduction in antibody and cellular responsiveness to a variety of common vaccines and investigate the underlying causes attributed to this phenomenon, such as inflammation and senescence. We will also discuss the impact of chronic viral infections on immune responses in both young and elderly patients, particularly those living with HIV, and how this affects vaccinations in these populations

The Long-Term Effects of Bereavement: A Longitudinal Study

The purpose of the present study was to examine the applicability of a model of bereavement to the long-term adjustment to loss. Based on Allen's (1990) model, it was predicted that the variables experienced competence, perceived resources, and the impact of the loss would contribute strongly to overall long-term bereavement adjustment. It was also predicted that time and multiple losses would impact adjustment to loss