Investigating novel approaches to the detection of virus neutralising antibodies to rabies and Rift Valley fever virus

Serosurveillance is a powerful tool fundamental to understanding infectious disease dynamics. The presence of virus neutralising antibody (VNAb) in sera is considered the best evidence of infection, or indeed vaccination, and the gold standard serological assay for their detection is the virus neutralisation test (VNT). However, VNTs are labour intensive, costly and time consuming. In addition, VNTs for the detection of antibodies to highly pathogenic viruses require the use of high containment facilities, restricting the application of these assays to the few laboratories with adequate facilities. As a result, robust serological data on such viruses are limited. In this thesis I develop novel VNTs for the detection of VNAb to two important, highly pathogenic, zoonotic viruses; rabies and Rift Valley fever virus (RVFV). The pseudotype-based neutralisation test developed in this study allows for the detection of rabies VNAb without the requirement for high containment facilities. This assay was utilised to investigate the presence of rabies VNAb in animals from a variety of ecological settings. In this thesis I present evidence of natural rabies infection in both domestic dogs and lions from rabies endemic settings. The assay was further used to investigate the kinetics of VNAb response to rabies vaccination in a cohort of free-roaming dogs. The RVFV neutralisation assay developed herein utilises a recombinant luciferase expressing RVFV, which allows for rapid, high-throughput serosurveillance of this important neglected pathogen. In this thesis I present evidence of RVFV infection in a variety of domestic and wildlife species from Northern Tanzania, in addition to the detection of low-level transmission of RVFV during interepidemic periods. Additionally, the investigation of a longitudinal cohort of domestic livestock also provided evidence of rapid waning of RVF VNAb following natural infection. Collectively, the serological data presented in this thesis are consistent with existing data in the literature generated using the gold standard VNTs. Increasing the availability of serological assays will allow the generation of robust serological data, which are imperative to enhancing our understanding of the complex, multi-host ecology of these two viruses.

Jung and Goddess: the significance of Jungian and post-Jungian theory to the development of the Western Goddess Movement

This study is concerned with the significance of Jungian and post-Jungian theory to the development of the contemporary Western Goddess Movement, which includes the various self-identified nature-based, Pagan, Goddess Feminism, Goddess Consciousness, Goddess Spirituality, Wicca, and Goddess-centred faith traditions that have seen a combined increase in Western adherents over the past five decades and share a common goal to claim Goddess as an active part of Western consciousness and faith traditions. The Western Goddess Movement has been strongly influenced by Jung’s thought, and by feminist revisions of Jungian Theory, sometimes interpreted idiosyncratically, but presented as a route to personal and spiritual transformation. The analysis examines ways in which women encounter Goddess through a process of Jungian Individuation and traces the development of Jungian and post-Jungian theories by identifying the key thinkers and central ideas that helped to shape the development of the Western Goddess Movement. It does so through a close reading and analysis of five biographical ‘rebirth’ memoirs published between 1981 and 1998: Christine Downing’s (1981) The Goddess: Mythological Images of the Feminine; Jean Shinoda Bolen’s (1994) Crossing to Avalon: A Woman’s Midlife Pilgrimage; Sue Monk Kidd’s (1996) The Dance of the Dissident Daughter: A Woman’s Journey from Christian Tradition to the Sacred Feminine; Margaret Starbird’s (1998) The Goddess in the Gospels: Reclaiming the Sacred Feminine; and Phyllis Curott’s (1998) Book of Shadows: A Modern Woman’s Journey into the Wisdom of Witchcraft and the Magic of the Goddess. These five memoirs reflect the diversity of the faith traditions in the Western Goddess Movement. The enquiry centres upon two parallel and complementary research threads: 1) critically examining the content of the memoirs in order to determine their contribution to the development of the Goddess Movement and 2) charting and sourcing the development of the major Jungian and post-Jungian theories championed in the memoirs in order to evaluate the significance of Jungian and post-Jungian thought in the Movement. The aim of this study was to gain a better understanding of the original research question: what is the significance of Jungian and post-Jungian theory for the development of the Western Goddess Movement? Each memoir is subjected to critical review of its intended audiences, its achievements, its functions and strengths, and its theoretical frameworks. Research results offered more than the experiences of five Western women, it also provided evidence to analyse the significance of Jungian and post-Jungian theory to the development of the Western Goddess Movement. The findings demonstrate the vital contributions of the analytical psychology of Carl Jung, and post-Jungians M Esther Harding, Erich Neumann, Christine Downing, E.C. Whitmont, and Jean Shinoda Bolen; the additional contributions of Sue Monk Kidd, Margaret Starbird, and Phyllis Curott, and exhibit Jungian and post-Jungian pathways to Goddess. Through a variety of approaches to Jungian categories, these memoirs constitute a literature of Individuation for the Western Goddess Movement.

Controlled surface nanotopography and oxygen plasma treatment of PEEK to improve cellular response

Poly(aryl-ether-ether-ketone) (PEEK) is a semi crystalline polymer which exhibits properties that make it an attractive choice for use as an implant material. It displays natural radiolucency, and MRI compatibility, as well as good chemical and sterilization resistance, both of which make it of particular interest in orthopaedic implants. However, PEEK has demonstrated poor cellular adhesion both in vitro and in vivo. This is problematic as implant surfaces that do not develop a layer of adhesive cells are at risk of undergoing fibrous encapsulation, which in turn leads to lack of a strong interface between the implant device and the patient tissue, which can in turn lead to failure of the implant and revision surgery . As incorporating nanotopography into a polymer surface has been demonstrated to be able to direct the differentiation behaviour of stem cells, a possible solution to PEEKs underlying issues with poor cellular response would be to incorporate specific nanoscale topography into the material surface through injection moulding, and then analysing if this is a viable method for addressing PEEKs issues with cellular response. In addition to nanoscale topography, the experimental PEEK surfaces were treated with oxygen plasma to address the underlying cytophobicity of the material. As this type of treatment has been documented to be capable of etching the PEEK surface, experiments were carried out to quantify the effect of this treatment, both on the ability of cells to adhere to the PEEK surface, as well as the effect it has upon the nanotopography present at the PEEK surface. The results demonstrated that there were a range of plasma treatments which would significantly improve the ability of cells to adhere to the PEEK surface without causing unacceptable damage to the nanotopography. Three different types of cells with osteogenic capacity were tested with the PEEK surfaces to gauge the ability of the topography to alter their behaviour: SAOS-2, osteoprogenitors and 271+ MSCs. Due to PEEKs material properties (it is non transparent, exhibits birefringence and is strongly autofluorescent) a number of histological techniques were used to investigate a number of different stages that take place in osteogenesis. The different cell types did display slightly different responses to the topographies. The SAOS-2 cells cultured on surfaces that had been plasma treated for 2 minutes at 200W had statistically significantly higher levels of von Kossa staining on the NSQ surface compared to the planar surface, and the same experiment employing alizarin red staining, showed a statistically significantly lower level of staining on the SQ surface compared to the planar surface. Using primary osteoprogenitor cells designed to look into if whether or not the presence of nanotopography effected the osteogenic response of these cells, we saw a lack of statistically significant difference produced by the surfaces investigated. By utilising HRP based immunostaining, we were able to investigate, in a quantitative fashion, the production of the two osteogenic markers osteopontin and osteocalcin by cells. When stained for osteocalcin, the SQ nanotopography had total percentage of the surface with stained material, average area and average perimeter all statistically significantly lower than the planar surface. For the cells that were stained for osteopontin, the SQ nanotopgraphy had a total percentage of the surface with stained material, average area and average perimeter all highly statistically significantly lower than those of the planar surface. Additionally, for this marker the NSQ nanotopography had average areas and average perimeters that were highly significantly higher than those of the planar surface. There were no significant differences for any of the values investigated for the 271+ MSC’s When plasma treatment was varied, the SAOS-2 cells demonstrated an overall trend i.e. increasing the energy of plasma treatment in turn leads to an increase in the overall percentage of staining. A similar experiment employing stem cells isolated from human bone marrow instead of SAOS-2 cells showed that for polycarbonate surfaces , used as a control, mineralization is statistically significantly higher on the NSQ nanopattern compared to the planar surface, whereas on the PEEK surfaces we observe the opposite trend i.e. the NSQ nanotopography having a statistically significantly lower amount of mineralization compared to the planar surface at the 200W 2min and 30W 1min plasma treatments. The standout trend from the PEEK results in this experiment was that the statistically significant differences on the PEEK substrates were clustered around the lower energy plasma treatments, which could suggest that the plasma treatment disrupted a function of the nanotopograhy which is why, as the energy increases, there are less statistically significant differences between the NSQ nanotopography and the Planar surface This thesis documents the response of a number of different types of cells to specific nanoscale topographies incorporated into the PEEK surface which had been treated with oxygen plasma. It outlines the development of a number of histological methods which measure different aspects of osteogenesis, and were selected to both work with PEEK, and produce quantitative results through the use of Cell Profiler. The methods that have been employed in this body of work would be of interest to other researchers working with this material, as well as those working with similarly autofluorescent materials.