Herpes simplex virus type-1 (HSV-1) and its closely related type-2 (HSV-2) viruses cause important clinical manifestations in humans including acute ocular disease and genital infections. These viruses establish latency in the trigeminal ganglionic and dorsal root neurons, respectively. Both viruses are widespread among humans and can frequently reactivate from latency causing disease. Currently, there are no vaccines available against herpes simplex viral infections. However, a number of promising vaccine approaches are being explored in pre-clinical investigations with few progressing to early phase clinical trials. Consensus research findings suggest that robust humoral and cellular immune responses may partially control the frequency of reactivation episodes and reduce clinical symptoms. Live-attenuated viral vaccines have long been considered as a viable option for generating robust and protective immune responses against viral pathogens. Varicella zoster virus (VZV) belongs to the same alphaherpesvirus subfamily with herpes simplex viruses. A live-attenuated VZV vaccine has been extensively used in a prophylactic and therapeutic approach to combat primary and recurrent VZV infection indicating that a similar vaccine approach may be feasible for HSVs. In this dissertation, we summarize pre-clinical approaches to HSV vaccine development and current efforts to test certain vaccine approaches in human clinical trials. Also, we discuss the potential advantages of using a safe, live-attenuated HSV-1 vaccine strain to protect against both HSV-1 and HSV-2 infections.

The well conserved Notch signalling pathway plays a crucial role in vascular development and physiology. Delta-like 4 (DLL4) and Jagged1 (JAG1) are two key notch ligands implicated in angiogenesis. Both ligands were shown to have opposite effects on vasculature. DLL4-Notch signalling inhibits sprouting resulting in fewer but better perfused blood vessels, promoting tumour growth. In contrast to DLL4, very little is known about JAG1- Notch signalling in tumour angiogenesis and its influence on tumour growth and progression. The overall aim of this work is to study the functional difference between DLL4 and JAG1-Notch signalling. The effects of murine DLL4 and murine JAG1 over-expression on tumour growth and angiogenesis was also investigated in a mouse U87 xenograft model. Firstly, the downstream target genes of DLL4 and JAG1-Notch signalling were established through microarray and QPCR. Angiogenic assays such as sprouting, network formation and migration assays were employed to study the functional effects of these two ligands in endothelial cells. The thesis firstly demonstrates that JAG1 has opposing effects on endothelial cells compared to DLL4 by increasing sprout coverage and network formation. JAG1 is less potent than DLL4 in stimulation of Notch target genes in primary endothelial cell (HUVEC) but both displayed equal potency in HMEC-1, an immortalised endothelial cell line. The growth of U87 cell lines which over-expressed murine DLL4 or murine JAG1 was slower compared to wild-type U87 cell line in vitro. JAG1- and DLL4- Notch signaling have different effects on vessel formation, which impacted on the tumour growth in vivo. Interestingly, tumours over-expressing mDLL4 had less but larger vessels compared to control, whereas mJAG1 produced more yet functional vessels; both tumours had significantly reduced pericyte coverage. Both U87 mDLL4 and mJAG1 over-expressing tumours showed increased resistance towards anti-VEGF therapy, compared to control tumours. Sensitivity to therapy was restored in combinational treatment with DBZ and bevacizumab. The mechanism behind the differential responsiveness of the Notch receptors to DLL4 or JAG1 ligands could either reflect modulation by fringes, a family of glycosyltransferases that regulate Notch signalling or by a positive feedback loop present for DLL4-Notch signalling only. Fringe was found to be abundantly expressed in endothelial cells and highly vascularised tumours. This work has highlighted some key novel differences between the two Notch Ligands.

While the major source of vitamin D is D₃ from ultraviolet exposure, some supplements supply D₂. The relative potency of vitamin D₂ versus vitamin D₃ remains controversial. The aims of the current study were, 1. To optimize the in vitro model, including use of cell lines, vitamin D concentrations, and outcome biomarkers. 2. To compare the potency of vitamin D₂ and D₃ metabolites on mouse and human bone cellular activity. 3. To explore the expression of VDR in osteoarthritic (OA) bone tissues as well as cellular responses to vitamin D₂ and D₃ metabolites ex-vivo. In mouse 2T3 osteoblasts, at physiological doses, both vitamin D₂ and D₃ metabolites increased ALP activity and mineralisation and up-regulated osteoblastic signature genes and proteins. At supra-physiological doses D₃ metabolites were more potent inhibitors of 2T3 function than D₂ metabolites. Although hBMS cell proliferation was inhibited by both 25(OH)D₂ and D₃, ALP activity was enhanced by both metabolites. However, 25(OH)D₃ was a more potent stimulator of ALP and mineralisation of hBMSCs. D₂ and D₃ equally stimulated expression of CX43 and PHEX markers in osteocytic cell lines. Immunohistochemistry of femoral heads showed much reduced VDR expression in OA osteocytes and osteoclasts, yet both 25(OH)D2 and D₃ increased OA-hBMSCs mineralisation more than non-OA-hBMSCs ex-vivo. While vitamin D₂ or D₃ increased mouse 2T3 osteoblastic activity at physiological doses, OA and non-OA hBMSCs differentiation was more responsive to 25(OH)D₃. Key bone cells such as osteocyte and osteoclasts expressed less VDR in OA. For the first time vitamin D₂ metabolites have been thoroughly examined and emerged as a potent stimulator of bone cell differentiation, at least in vitro. Vitamin D₃ in contrast is confirmed as highly potent in bone cells, but with toxicity at much lower doses than D₂.

After the emergence of Zika virus (ZIKV) in the Americas in 2015, ZIKV infection was associated for the first time since its discovery with severe symptoms in both adults and congenital cases, including neurological, ocular, and developmental manifestations. Previous ZIKV circulation in Africa and Southeast Asia has been characterized by mild symptoms and small-scale case-counts. It is unclear whether the unprecedented size and severity of the ZIKV outbreak in the Americas are the consequence of a change in the virus, different background flaviviral immunity in the population, or a reporting issue. In addition, ZIKV has been shown to be transmitted through sexual contact, and the shedding of ZIKV from various bodily fluids in both humans and in vivo models suggests that other potential routes of transmission exist. We present here two mouse models that can be used to further investigate ZIKV pathogenesis, transmission, and immune response. Mice lacking interferon regulatory factors 3 and 7 (IRF 3/7 DKO) supported robust infection with the prototype MR766 strain from Uganda, while maintaining a 72% survival rate, and recapitulated symptoms and tissue lesions associated to infection with American ZIKV isolates in humans and other in vivo models. The MR766 strain was capable of causing retinal lesions and viral RNA shedding from the conjunctival fluid, hitherto unreported to be caused by an African strain. Further, ZIKV was visualized in the seminal fluid co-localized with infected epithelial epididymal cells, suggesting a possible cellular component of sexual transmission. Immunocompetent C57BL/6 mice, although not susceptible to ZIKV, were capable of mounting a robust antibody response that strongly neutralized ZIKV and was also able to cross-neutralize DENV2. We further report that homologous re-exposure with ZIKV in C57BL/6 mice reduced the DENV2 cross-neutralizing capability of the antibody population, while at the same time increasing enhancement of DENV2 infection. We conclude that ZIKV strains of the African and Asian lineages share a similar pathogenesis, suggesting that the increased severity of symptoms is unlikely to be due to a change in the virus. We also show that re-exposure to ZIKV can alter the antibody response to increase the risk of heterologous infection.

Early-onset spinal deformity (EOSD) is characterised by detection of spinal deformity (scoliosis, kyphosis or multi-planar) in children at aged less than five years. The common causes could be classified under congenital, neuromuscular, syndromic and idiopathic etiologies. Early treatment is paramount in preventing rigid, severe and progressive deformities that can cause pulmonary compromise. The inability of lungs and thoracic cage to support normal ventilation at rest constitutes the pathophysiology in manifestation of thoracic insufficiency syndrome (TIS). The treatment options have evolved from observation, serial casting, bracing to surgery. Early definitive spinal fusion is now obsolete and contra-indicated for EOSD. Growing rods (submuscular or subfascial) continues to be the standard of care in treating these challenging deformities. Vertebral expandable prosthetic titanium rib (VEPTR) continues to be an attractive option for TIS but is fraught with high complication rate. I hereby present a theme on EOSD with a set of thirty three peer reviewed indexed publications and one surgical patent in treating such challenging conditions highlighting my original contribution as Consultant spinal surgeon spanning over two decades. My pioneering and ground-breaking research that has shaped the surgical management of EOSD and helped define 'standard of care' is presented. My novel and innovative concept of treating EOSD using magnet driven growing rod (MdGR) along with its preliminary results is discussed. MdGRs are an attractive alternative in eliminating need for repetitive anaesthesia facilitating normal cognitive development in comparison to growing rods (CGRs). They also improve pulmonary function in neuromuscular scoliosis. A brief one page summary of all my indexed publications with comments on originality and how they contributed to spinal surgery is enclosed at the end of each chapter. My current research update on MdGR project at Royal National Orthopaedic Hospital (Stanmore), my surgical patent National institute of clinical excellence (NICE) position statement on MdGRs and clinical guidance documents are attached appendices I - III.

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that circulates predominantly in tropical and subtropical regions. Infection results in severe debilitating polyarthralgia during the acute phase of disease, and reports suggest that chronic arthralgia lasting months to years after the initial infection can occur. More severe and prolonged disease has been associated with pre-existing joint disease, though this has not been experimentally examined. In the research presented herein, two established mouse models (adult IRF 3/7 -/- -/- and wild-type C57BL/6J mice) were utilized to characterize CHIKV-associated bone and joint disease and evaluate its impact on the progression of pre-existing osteoarthritis (OA) utilizing histopathology, μCT, and serology. During acute stages of the disease, CHIKV infection resulted in synovitis, cartilage necrosis, and periosteal necrosis or periostitis. Additionally, IRF mice had severe ischemic bone marrow necrosis, and C57BL/6J mice developed periosteal new bone proliferation. During chronic stages of disease (90 DPI), there was ongoing and progressive synovitis, tendonitis, enthesitis, and cartilage damage, though periostitis and periosteal bone proliferation had resolved. Infection with CHIKV in mice with pre-existing OA had no significant impact on total synovitis scores or chondrocyte cell death, but caused a decrease in volume of osteophytes and subchondral bone, as compared to OA alone. Serology results in both the footpad and intra-articular C57BL/6J models indicated there were alterations in RANKL and OPG associated with CHIKV infection, demonstrating potential changes in bone dynamics. The current experiments demonstrated novel lesions of CHIKV-associated bone and joint disease that have important ramifications for the treatment and prevention of disease. Periosteal bone proliferation associated with CHIKV is a potentially painful but reversible process, whereas articular cartilage damage is progressive and represents a potential mechanism for chronic CHIKV-associated joint disease. Additionally, the alterations in bone associated with CHIKV infection and pre-existing OA, in conjunction with changes in the RANKL-OPG pathways, could have significance in clinical monitoring of OA and treatment choices in individuals with concurrent diseases. Future studies would help determine if RANKL and OPG levels could be useful for tracking disease progression in people and establish the long-term effects of CHIKV infection on OA progression.

An eating disorder is a psychological disorder that affects one’s eating habits. According to a 2011 study, approximately 20 million women and 10 million men in the United States suffer from a clinical eating disorder at some point in their lives (Wade, Keski-Rahkonen, & Hudson, 2011). Eating disorders include anorexia nervosa, bulimia nervosa, binge eating disorder, and other disorders (DSM V, 2013). Recent research has indicated that the prevalence of anorexia nervosa is 0.3% in males, 0.9% in females, and 0.3% in adolescents (Hoek, 2006). In 2007, the first national survey to include eating disorders found the prevalence of bulimia nervosa to be 1.5% in the United States (Hudson, Hiripi, Kessler, 2007). The fifth edition of Diagnostic and Statistical Manual of Mental Disorders (2013) was revised to include binge eating disorder, which has a prevalence rate of 2.6% in white women and 4.5% in African American women (DSM V, 2013; Pike, Dohm, Striegel-Moore, & Fairburn, 2001; Striegel-Moore, Wilfley, & Pike, 2000). Though overall prevalence rates are low, eating disorders impact millions of Americans each year, and eating disorders have the highest mortality rate of all psychiatric disorders(Neumarker, 2000). Music therapy, defined as the use of musical interventions to address non-musical goals, has its roots in mental health care (Silverman, 2015). While music therapy is one of the treatment options for individuals with eating disorders, very little research is available regarding music therapy with this population (Hilliard, 2001). This manual provides an overview of music therapy for the treatment of eating disorders with the purpose of better equipping music therapists with the knowledge and tools to serve the growing eating disorder population. / A Thesis submitted to the College of Music in partial fulfillment of the requirement for the degree of Master of Music. / Summer Semester 2017. / June 21, 2017. / eating disorders, music therapy / Includes bibliographical references. / Lori Gooding, Professor Directing Thesis; Jayne Standley, Committee Member; Diane Gregory, Committee Member.

Researchers that investigate the media's coverage of health have historically relied on keyword searches to retrieve relevant health news coverage, and manual content analysis methods to categorize and score health news text. These methods are problematic. Manual content analysis methods are labor intensive, time consuming, and inherently subjective because they rely on human coders to review, score, and annotate content. Retrieving relevant health news coverage using keywords can be challenging because manually defining an optimal keyword query, especially for complex health topics and media analysis concepts, can be very difficult, and the optimal query may vary based on when the news was published, the type of news published, and the target audience of the news coverage. This dissertation research investigated computational methods that can assist health news investigators by facilitating these tasks. The first step was to identify the research methods currently used by investigators, and the research questions and health topics researchers tend to investigate. To capture this information an extensive literature review of health news analyses was performed. No literature review of this type and scope could be found in the research literature. This review confirmed that researchers overwhelmingly rely on manual content analysis methods to analyze the text of health news coverage, and on the use of keyword searching to identify relevant health news articles. To investigate the use of computational methods for facilitating these tasks, classifiers that categorize health news on relevance to the topic of obesity, and on their news framing were developed and evaluated. The obesity news classifier developed for this dissertation outperformed alternative methods, including searching based on keyword appearance. Classifying on the framing of health news proved to be a more difficult task. The news framing classifiers performed well, but the results suggest that the underlying features of health news coverage that contribute to the framing of health news are a richer and more useful source of framing information rather than binary news framing classifications. The third step in this dissertation was to use the findings of the literature review and the classifier studies to design the SalientHealthNews system. The purpose of SalientHealthNews is to facilitate the use of computational and data mining techniques for health news investigation, hypothesis testing, and hypothesis generation. To illustrate the use of SalientHealthNews' features and algorithms, it was used to generate preliminary data for a study investigating how framing features vary in health and obesity news coverage that discusses populations with health disparities. This research contributes to the study of the media's coverage of health by providing a detailed description of how health news is studied and what health news topics are investigated, then by demonstrating that certain tasks performed in health news analyses can be facilitated by computational methods, and lastly by describing the design of a system that will facilitate the use of computational and data mining techniques for the study of health news. These contributions should further the study of health news by expanding the methods available to health news analysis researchers. This will lead to researchers being better equipped to accurately and consistently evaluate the media's coverage of health. Knowledge of the quality of health news coverage should in turn lead to better informed health journalists, healthcare providers, and healthcare consumers, ultimately improving individual and public health.

Sound arriving at the ear causes the vibration of the sensory tissues, including the basilar membrane (BM), inside the cochlea and, in turn, leads to inner hair cell excitation and auditory nerve fiber (ANF) responses. The goal of this study is to better understand the mechanics of inner hair cell excitation which leads to hearing. BM motion and ANF tuning are generally very similar, but the ANF had appeared to be unresponsive to a plateau mode of BM motion that occurs at frequencies above an ANF's characteristic frequency (CF). We recorded ANF responses from the gerbil, concentrating on this supra-CF region. We observed a supra-CF plateau in ANF responses at high stimulus level, indicating that the plateau mode of BM motion can be excitatory. Quantitative aspects of our findings suggest that the differential longitudinal motion that occurs within the traveling wave but not the plateau mode increases the sensitivity of inner hair cell excitation. The main findings of this study include: The detection of the plateau threshold within the supra-CF region of the ANF tuning curve. A larger BM motion was necessary for an ANF to reach a threshold response within the plateau region than the traveling wave region, based on the previous lack of ANF plateau threshold detection and a comparison to the BM plateau levels in the literature. Stimuli used in this study, even though unnaturally high in level, advanced our understanding of cochlear mechanics. However, at high sound pressure levels used, the middle ear generated subharmonic distortions that could produce confounding effects in the plateau responses. Hence, we also characterized the subharmonics and were able to rule out the possibility that they were solely responsible for the plateau responses we observed.

Atherosclerosis, the primary cause of heart attack, stroke, and peripheral vascular disease, is genetically complex and the genes that confer cardiovascular risk remain largely unknown. Attempts to map common atherosclerosis susceptibility loci in humans have resulted in limited success. Mouse models provide an excellent tool for dissecting genetic complexity,and testing the role of candidate genes and pathways. While recent genome-wide association studies in humans have identified multiple genetic variants associated with atherosclerosis related traits- such as circulating lipid levels and hypertension- as well as myocardial infarction and stroke, few of the causal variants or underlying mechanisms are known. The work presented in this thesis identifies genetic loci contributing to atherosclerosis susceptibility in a murine model and, based on mechanistic data obtained in mouse, strongly suggests a mechanism underlying a coronary heart disease susceptibility locus on chromosome 9p21 identified by genome-wide association studies in humans. In this study we have used congenic mice that were previously constructed to isolate Athsq1, an atherosclerosis susceptibility locus identified in a cross between MOLF/Ei and C57BL/6J-Ldlr-/-. Atherosclerosis studies performed in congenic mice carrying the MOLF-derived susceptibility allele from chromosome 4, in the C57BL/6J-Ldlr-/- background, confirmed the existence of the Athsq1 locus and BM-derived cells were shown to play an important role. Refined mapping of the Athsq1 locus identified at least two susceptibility loci within the interval. A proximal locus was narrowed to a region containing 8-21 genes, including the region of homology with the human coronary heart disease risk interval on chromosome 9p21. Interestingly, mRNA expression analyses in macrophages revealed markedly decreased Cdkn2a transcript expression in cells derived from congenic mice compared to the controls. Cdkn2a is a candidate locus associated with the 9p21 locus in humans. The potential role of bone marrow -derived Cdkn2a expression in atherogenesis was tested using a bone marrow transplantation approach with Cdkn2a-deficient donor cells. Mice receiving Cdkn2a+/-deficient bone marrow exhibited accelerated atherosclerosis, increased inflammatory monocyte subsets and increased monocyte/macrophage proliferation compared to the controls. This study provides a mechanistic link between decreasedCdkn2aexpression, increased monocytes/macrophages proliferation with the expansion of an inflammatory monocyte subset and increased atherosclerosis. Together, these data illustrate the utilization of mouse models as tools to elucidate causal gene(s)/loci and potential pathophysiologicmechanisms underlying genome-wide association studies-identified loci for human disease.