The cochlea is the organ for sound reception. Mammals place varied functional demands on their sense of hearing to meet the requirements of a broad range of ecological niches and diverse behaviours. However, documenting potentially related adaptations of the cochlea to eco-behavioural traits is difficult due to its complex geometry. The present study aims to determine whether the bony cochlea carries eco-behavioural traits that can be used to contextualize our understanding of the fossil record and evolutionary transitions. This study also includes work on ontogenetic changes since these can yield important insights into evolutionary processes resulting in differences of the adult phenotypes. Advanced techniques in micro-CT imaging, 3D image visualization, geometric morphometrics and statistical methods were used to study morphological variations of the bony cochlea across 45 adult eutherian species. Also, the same set of techniques was used to study 12 human fetal (approximately four to nine months of gestation) cochleae in comparison with five adult cochleae. Results revealed that there was a considerable range of variation in form of the mammalian bony cochlea. Potential links between the bony cochlear morphology and hearing, ecology and behaviour were found. Dimensions of the bony cochlea may be indicative of the eco-behavioural niche that a mammal occupies; e.g., fewer than two spiral turns is associated with obligate marine species. Rodents also showed remarkable variation in the cochlear morphology, more so than any other group of mammals studied, reflecting their diverse eco-behavioural traits. Results from the human developmental study showed that whilst the general coiled shape was achieved at the midgestational age, there was size related morphological change during the postnatal period. The round window size reached mature state prior to birth, by approximately the second trimester, whereas the oval window continued to change in size after birth. The postnatal enlargement may be determined by functional requirements of air-borne hearing, particularly with respect to frequency range and sensitivity.

It is hypothesised that there are distinct mechanisms involved in cartilage ageing and disease which can be determined using next-generation technologies including mass spectrometry and RNA-sequencing. The aims of this thesis were to firstly characterise molecular mechanisms associated with age-related and arthritis-related changes in cartilage gene and protein signatures. Secondly the thesis developed new techniques to identify novel cleavage sites in matrix proteins and to quantify some known proteolytic events in articular cartilage using mass spectrometry-based proteomics platforms. Finally the levels of key proteinases and their inhibitors involved in the pathogenesis of OA were measured using mass spectrometry. Osteoarthritis (OA) is an extremely common cause of morbidity in both man and animals. OA involves the biomechanical failure of articular cartilage, together with changes in the subchondral bone and inflammation of the joints and leads to a variety of symptoms including pain, stiffness and reduced mobility. Age is an important factor in the development of OA and represents a huge challenge for society as whilst life span increases, the quality of life faced by an ageing population is often poor. Articular cartilage is susceptible to age-related diseases such as OA, but it is not an inevitable result of ageing and is a consequence of a complex inter-relationship between age and further predisposing factors. There have been major advances in technologies used to interrogate proteins and genes due to genome sequencing enabling gene and protein sequences to be determined. These ‘next-generation technologies’ include mass spectrometry (MS) and next-generation sequencing. This thesis has used these technologies in an attempt to address important questions relating to cartilage ageing and disease. The use of an inflammatory model of early OA in equine and human cartilage enabled the discovery and quantification of important proteins and pathways involved, using relative and absolute mass spectrometry techniques. In the equine secretome pathway enrichment analyses confirmed the up-regulation of glycolytic proteins. The novel proteins clathrin and LIM and SH3 domain protein-1 were identified for the first time in cartilage proteomics. QconCAT technology and gene expression analysis enabled normal and OA cartilage extract to be interrogated. Absolute quantification values were demonstrated for the first time for aggrecan; first and third globular domains, biglycan, cartilage oligomeric matrix protein, decorin and fibromodulin. Whilst a novel MS based technique enabled previously identified and novel extracellular matrix cleavage sites derived from matrix metalloproteinase 3 and a disintegrin and metalloproteinase with thrombospondin motifs 4 digestion of cartilage to be determined. Some of these sites of degradation were also evident in OA but not normal cartilage using matrix assisted laser desorption ionization imaging MS (MALDI-IMS). Tentative markers of OA and ageing cartilage were also demonstrated. Finally an RNA sequencing study on ageing equine cartilage found an age-related failure of matrix, anabolic and catabolic cartilage factors together with a reduction in Wnt signalling. This thesis developed novel proteomic methodologies to identify and quantify distinct differences between cartilage ageing and disease. Several proteins not previously described in cartilage were identified. In addition many novel cartilage degradation products were identified and age-related peptides were visualised in cartilage for the first time.

Background: The ICD is cornerstone in the treatment of life-threatening arrhythmias, although there is growing concern that dying patients are receiving multiple futile shocks. Expert guideline recommendations regarding ICD deactivation are not implemented in practise and ICD deactivation is rarely discussed. This limits patients' and carers' knowledge and choice at end-of-life. Aim: To explore perspectives of patients, carers and professionals regarding ICD deactivation and to examine the impact these have on clinical judgements about end-of-life management. Methods: Sequential exploratory mixed methods design incorporating two phases. • Phase One: Data from a systematic review of literature, case studies and retrospective case note review were synthesised and used to generate nine independent variables. • Phase Two: Variables were randomly manipulated and embedded within vignettes of a factorial survey disseminated to UK and Irish professionals. Results: Phase One data confirmed pre-implantation information on ICD's functionality and possible deactivation was inadequate. Patients' and professionals' held a positive perception of the ICD and were reluctant to discuss deactivation until death was imminent. Most patients wanted involvement in critical discussions, although agreed the decision concerning deactivation should be made by their cardiologist, without burdening family. Carers' were kept uninformed unless the patient became cognitively impaired. Phase Two: 534 vignettes were completed by 89 professionals (22 Cardiologists, 57 Nurses, 10 Clinical Physiologists). Nurses were more likely to favour a pre-implantation discussion than cardiologists, although all groups agreed the subject of deactivation should be broached when death was imminent. Clinical indicators of heart failure severity (NYHA IV) and diagnosis of bowel cancer increased the likelihood of this discussion. All groups felt deactivation was warranted when the patient experienced multiple shocks, however data from Phase One found no evidence of this occurring. Professionals in post for at least six years were most confident in clinical decision-making. Conclusions: These data highlight missed opportunities to involve patients in shared decision-making, with the majority of professionals reluctant to discuss deactivation. Lack of pre-implantation information compromised patients' knowledge and restricted informed decision-making to last days of life. Professionals' relied on their intuitive judgement rather than evidence-based guidance. Data extends the factorial survey methodology and provides direction to improve end-of-life care for patients with an ICD.

No description available.

Kidney transplantation is a life transforming procedure which substantially improves the quality and quantity of life of individuals with end stage renal disease. The short term outcomes following kidney transplantation are now excellent but recipients continue to experience significant long term morbidity. This thesis investigates the clinical, genetic and epigenetic factors which influence long term graft and recipient outcomes after kidney transplantation with a particular focus on the major, life limiting complications. Chapter 3 investigates the impact of DNA methylation and single nucleotide polymorphisms in a candidate gene on death censored graft survival. In addition, the possibility of anticipating death censored graft survival from a recipients clinical course is explored. In Chapter 4, DNA methylation signatures associated with recipient survival and cardiovascular disease are investigated long with the genetic basis of new onset diabetes after transplantation and the national incidence of non melanoma skin cancer. Chapter 5 focuses on topical issues in kidney transplantation; the impact of obesity on graft and recipient outcomes and of graft failure on recipient survival are studied. Finally I the characteristics and clinical course of recipients with ultra long term graft survival are described in Chapter 6.

No description available.

Quality of Life (QOL) amongst transplant patients has become an issue as a means to optimize the use of scarce resources and improve outcome. The relationship between QOL, medical compliance, psychosocial states and QOL is presented as follows: Patient empowerment: Patient empowerment and compliance with medications in patients who experienced late acute rejection, was assessed using the Long-term Medication Behaviour Scale (LTMBS-scale). We demonstrated a relationship between late acute rejection and low confidence in taking medication and items relating to physical and psychological symptoms. Effect of psychotherapeutic program on emotional states after kidney transplantation: Recurring emotional states as recalled by patients during psychotherapy sessions (e.g. loss of QOL) were analysed and used to formulate effective group and individual psychotherapy intervention. The Beck Depression Inventory (BDI) was utilised as a measure of change before, during and after treatment. There was significant improvement in psychological states after therapy contemplated as follows, (i) fear of rejection, (ii) feelings of paradoxical loss (iii) psychological integration of the transplant. Randomised controlled study to determine the efficacy of individual or group psychotherapy amongst kidney transplant patients: Analysis of group and individual psychotherapy in recipients of kidney transplants versus control patients was conducted. Improvement appeared to be more significant in the individual therapy compared to group therapy (p= 0.01). In both the individual and group therapy arm, lowering of scores was progressive and sustained (p=O.OI). However, individual therapy resulted in the lowest BDI scores at the end of treatment period. Coming to terms with the 'imagined past': After a successful transplant, patients may present with feelings of paradoxical loss (e.g. grief or low mood). We propose that paradoxical loss should be considered in terms of an imagined past (loss of life goals which have never been actualised as a result of chronic illness). Feelings of loss present an obstacle to a good QOL after transplantation as low mood may lead to ambivalence and subsequently to non-compliance.

The Notch signaling pathway is critically involved in cell fate decisions during skin development and homeostasis. In the present study an in vivo model (seven-week-old male, C57BL/6) was used to elucidate the role of epidermal Notch activation in the healing of full-thickness dermal wound. Immunolocalization of activated Notch1, Notch2 and Jagged1 was performed during skin wound healing in vivo in mice. The expression level of Notch1, Notch 2 and Jagged1 in the C57BL/6 skin was examined by quantitative PCR (qPCR). This study shows that skin injury rapidly actives Notch signaling in the epidermis. Epidermal forced Notch1 activation results in a Jagged1 dependent Notch2 activation in epidermis. Data presented in this thesis also demonstrate that the prolonged epidermal Notch activation via a 4-hydroxy-tamoxifen-inducible transgene before and after wounding caused an over-activation of Notch during early stages of the healing of full-thickness dermal wounds. The expression level of Notch1, Notch2 and the inflammatory Notch related genes, in the transgenic wounded skin was examined by quantitative PCR (qPCR). The phenotypes and morphology of the transgenic skin were compared with that of wild type (WT) controls. The skin response to wound healing was studied by H&E staining at the microscopic level at 2, 5 and 8 days post injury. These data show that the prolonged epidermal Notch activity may do more harm than good in terms of an increased inflammatory response at the wound site. These data suggest that Notch plays an important role in the early stages of the skin wound healing process, a finding that has implications in wound inflammatory responses. This thesis also examines skin wound healing in different anatomical locations on seven-week-old male, C57BL/6 mice model. 4mm full thickness dermal wounds were made at different anatomic regions; upper posterior, middle to posterior and posterior-most (caudal). These data showed that lower body wounds healed significantly better than those in the upper posterior, or middle to posterior of the mouse back skin. The data presented here show that anatomical location is important in wound healing responses, as reflected by differences in keratinocyte proliferation and differentiation.

No description available.

No description available.