Research has suggested that a bi-directional relationship exists between sleep disruption and depression. Not only is poor sleep a commonly reported symptom in those with depression, some aspects of sleep have also been shown to predict the onset of depression. Despite sleep problems being a commonly reported occurrence throughout the perinatal period, the field of perinatal sleep research remains in its relative infancy. However, recent studies suggest that sleep disturbances during this time may increase the risk of developing postpartum depression. Currently, research in this area is limited by studies that have failed to control for depressive symptoms at baseline, relied upon subjective, often retrospective, measures of sleep, and have only measured symptoms of postpartum depression in the early postpartum period. Few studies have used polysomnography, considered the ‘gold standard’ of sleep, and no studies to date have specifically compared the relationship between subjective and objective sleep. Therefore, the major aim of this thesis was to gain a better understanding of the specific aspects of sleep that were most relevant to postpartum depression. In order to address this aim, studies were carried out to: explore the aspects of sleep most relevant to major depressive disorder; examine differences in sleep between pregnant and non-pregnant women; investigate the relationships between subjective and objective measures of sleep; explore longitudinal changes in sleep, fatigue and depression throughout the perinatal period, and finally; examine which aspects of sleep at which time-point were most relevant to the development of postpartum depression. Overall this thesis found that women experience significant changes to their sleep throughout the perinatal period. While the sleep of third trimester women is considerably poorer than that of non-pregnant women (both objectively and subjectively), the most significant changes occur in the transition between late pregnancy and the early postpartum period. Furthermore, increased amounts of sleep and reports of difficulty falling asleep during late pregnancy predicted the development of postpartum depressive symptoms. This suggests that certain aspects of sleep during late pregnancy may serve as markers for women at risk of developing postpartum depression.

'... we treat them all the same'. This study aimed to explore the experiences of patients, lay carers and health and social care staff of care received by older people with dementia in the acute hospital setting. In view of the ageing population, an increasing prevalence of dementia and the emerging dignity agenda this is a particularly topical subject. Four elements are included in this thesis: a literature review, a research study, a practice development project and an integrative review. A literature review sets person-focused research in the context of dementia research as a whole. It also identifies ways in which studies have been conducted in an ethical and meaningful manner. An ethnographic approach was used in the research study to collect data through observation, conversations and interviews. Findings portray a bleak picture of the care of this vulnerable group. Patients demonstrated, through words and actions, how difficult they found the experience. Lack of communication was an issue as was the obvious distress caused by delivery of personal care. Lay carers were relatively uncritical although this may have been due, in part, to the recruitment process. Staff generally appeared to have good intention. Most stated that they had received little or no preparation or education in dementia care. Many staff functioned almost entirely within thebed and body'framework. They frequently avoided communication with patients. Staff worked in a deeply embedded habitus, in which they appeared not to really think about what they were doing. They demonstrated a lack of empathy with patients. The concept that staff were working in a 'switched off mode and lacked empathy called for a practice development project that engaged them on both cognitive and emotional levels. A practice development project based on the philosophies of confluent education and situated learning was implemented. Initial evaluation has demonstrated some tangible changes in practice. An integrative review draws these elements together into a coherent whole. Potential contributions to the body of knowledge are acknowledged, as are limitations of the work. This study has shown that people with dementia, even those at an advanced stage and with superimposed physical illness, can be engaged in research that is both ethical and meaningful. It has shown that improvements in practice are possible. They need to be underpinned by a belief in the personhood of staff as well as patients. Areas for further research and practice development in this vital subject have been identified.

Day case surgery is expanding in the UK and is the favoured approach to elective surgery by the Government and patients alike. However studies have revealed patients' experience unacceptable postoperative pain when they return home after day surgery, leading to a variety of negative consequences, stemming many years, affecting many lives, with emotional and financial cost. It is imperative that pain is adequately controlled following day surgery to reduce these consequences and ensure the potential of day surgery is reached. Previous research has investigated barriers to pain management in this area, one barrier that has received little attention is that posed by the patient, and it has been suggested that patients may not utilising their analgesics appropriately with papers calling for further research in this area. Employing Interpretative Phenomenological Analysis (IPA) this study explored patients' use of analgesics on returning home following day surgery. Using IPA for analysis, interviews with twenty eight patients revealed many to avoid analgesics enduring severe postoperative pain during their recovery, and provided new understanding showing patients' use of analgesics to be as a result of a complex intentional decision making process based on a matrix of beliefs they held surrounding pain, analgesics and day surgery. These beliefs were found to be influenced by past experience, and cultural context, with this research being the first to identify many of these beliefs and make further sense of them by producing an explanatory framework illustrating how they exert their influence upon patients' decisions regarding analgesic use. One implication of these findings is that day surgery is not as straightforward as suggested, and simply providing patients pain management information and effective analgesics underestimates the complexity of the patient's experience when they return home. Further research is now required to identify alternative ways to reduce pain following day case surgery. One recommendation is to overcome erroneous beliefs held by patients. In particular the explanatory framework produced by this research provides a unique insight into the mechanism by which these beliefs may exert their influence upon patients' analgesic use, and may prove a useful tool to achieving this, overcoming pain and its negative consequences, paving the way for day case surgery to reach its full potential.

Primary aetiologic factors of diabetic foot disease include peripheral neuropathy and peripheral vascular disease. Assessment of circulation, neuropathy, and foot pressure is employed routinely to determine the risk of foot ulceration in the patient with diabetes mellitus. Routine neuropathic evaluation includes assessment of sensory loss in the plantar skin of the foot using both the Semmes Weinstein monofilament and the biothesiometer. Progressive degeneration of sensory nerve pathways is thought to affect thermoreceptors and mechanoreceptors. However, thermological measurements of the foot to assess responses to thermal stimuli and cutaneous thermal discrimination threshold are relatively uncommon. Recent improvements in liquid crystal technology (LCT) including insensitivity to pressure, faster response times, lower cost and fast image acquisition offer potential for routine thermographic assessment of the diabetic foot. The present study was designed to evaluate if an association exists between abnormal plantar thermal images and sensory loss under conditions of normal loading. The system comprises a robust measurement platform, thermochromic liquid crystal polyester sheet (TLC), instrumentation and analysis software. In vitro calibration was performed to characterise three physical forms of TLC on the basis of linearity, hysteresis, pressure sensitivity and response time. An in vivo pilot evaluation study of the system was performed using three sub-groups (i) neuropathic diabetic (n=30), (ii) non neuropathic diabetic (n=30) and (iii) a healthy control group (n=30). The principal results of this study indicate raised plantar temperatures for the diabetic groups at baseline and post stress relative to the control group. Furthermore, poor recovery response to thermal stimulus in the neuropathic diabetic group suggests degeneration of thermoreceptors. Thus by assessing the thermal parameters at the same sites as that of sensory testing, the new LCT based approach appears capable of providing an alternative confirmation of clinical neuropathy and offers potential as an improved method compared to existing techniques.

Spinal and back deformities can lead to pain and discomfort, disrupting productivity, and may require prolonged treatment. The conventional method of assessing and monitoring tile de-formity using radiographs has known radiation hazards. An alternative approach for monitoring the deformity is to base the assessment on the shape of back surface. Though three-dimensional data acquisition methods exist, techniques to extract relevant information for clinical use have not been widely developed. Thi's thesis presentsthe content and progression of research into automated analysis and visu-alization of three-dimensional laser scans of the human back. Using mathematical shape analysis, methods have been developed to compute stable curvature of the back surface and to detect the anatomic landmarks from the curvature maps. Compared with manual palpation, the landmarks have been detected to within accuracy of 1.15mm and precision of 0.8111m.Based on the detected spinous process landmarks, the back midline which is the closest surface approximation of the spine, has been derived using constrained polynomial fitting and statistical techniques. Three-dimensional geometric measurementsbasedon the midline were then corn-puted to quantify the deformity. Visualization plays a crucial role in back shape analysis since it enables the exploration of back deformities without the need for physical manipulation of the subject. In the third phase,various visualization techniques have been developed, namely, continuous and discrete colour maps, contour maps and three-dimensional views. In the last phase of the research,a software system has been developed for automating the tasks involved in analysing, visualizing and quantifying of the back shape. The novel aspectsof this research lie in the development of effective noise smoothing methods for stable curvature computation; improved shape analysis and landmark detection algorithm; effective techniques for visualizing the shape of the back; derivation of the back midline using constrained polynomials and computation of three dimensional surface measurements. �

Innate immunity depends on pattern recognition receptors, which recognize pathogen associated molecular patterns (PAMPS), such as Toll-like receptor 4 (TLR4), which detects the gram-negative bacterial toxin, lipopolysaccharide. Engagement of TLR4 by LPS sets off a cascade ending in the activation of pro-inflammatory cytokines and interferon-β (IFN-β) which alerts the host to the infection. However, these responses can be mal-adaptive, especially in the context of bacterial sepsis, where a "cytokine storm" results in death of the host. Pharmacological modulation of these responses may therefore be a promising treatment modality. Inhibition of classic pro-inflammatory cytokines such as IL-1β and TNF-α has been the (largely unfruitful) focus of much research. However it has recently emerged that mice with defects in type I IFN signaling are also substantially resistant to challenge with endotoxin. We therefore wish to investigate pharmacological inhibition of IFN signaling as a potential means to control sepsis. We analyzed the effects of Trichostatin A (TSA) and Suberoylanilide hydroxamic acid (SAHA) (both broad spectrum HDAC inhibitors) and ST-2-92 (HDAC6 specific inhibitor) on IFN regulation and endotoxic shock. We created an in vivo mouse model for this treatment with TSA and SAHA (which are well tolerated in mouse and human) to look for possible alteration in the survival rate following endotoxin challenge. We as well as others found that treatment with SAHA (50mg/kg) significantly improves survival rate. We also characterized in-vitro modulation of IFN responses through SAHA by mouse DNA microarray. We noticed a decreased expression of many innate immune regulated genes in the SAHA and LPS treated condition compared to the LPS treatment alone. Additionally we observed a decrease in protein levels of IFN-β IL-1β, IL-6, IL-12p40, RANTES and TNF-α in cell culture supernatants treated with SAHA or ST-2-92 and LPS compared to LPS only treatment. These results show the ability of broad spectrum HDACi through SAHA to increase mouse survival following LPS challenge as well as modulate the induction of innate immune responsive genes in vitro. Furthermore we have shown that HDAC specific inhibition through ST-2-92 can decrease pro-inflammatory transcript as well as protein levels.

In the United States, blood product availability is dependent entirely on donations from volunteer blood donors. Current trends in blood collection and utilization raise concerns about the ability to meet future demands for blood products. At a time of high demand and deferrals, the Armed Services Blood Program has been unable to meet its requirements for blood and consequently needs to purchase blood from civilian agencies to meet the dual demands of the military community at home, as well as those deployed around the world. This creates a need to better understand the military blood donor in an effort to recruit and retain those relative few who are willing and eligible to donate. The purpose of this survey-based research is to characterize the demographics of the military blood donor and to understand, through descriptive and regression analysis, the relationship between a donor’s attitude, subjective norms, and perceived behavioral control and their intention to donate blood again in the next six months. The study uses the framework of the theory of planned behavior to design the survey and evaluate relationships between the theoretical constructs. Descriptive analysis results of the sample demographics describe the typical respondent as White, young, male, and a junior service member. Results of the multivariate regression analysis showed that a respondent’s attitude toward blood donation and perceived behavioral control over donating blood were positively related to their intention to donate again in the next six months. However, contrary to the theory, there was no statistically significant relationship between the influence of subjective norms and intention to donate again. This study is the first to apply a theoretical framework to identify those factors that influence a military blood donor to donate blood. Further, it has taken steps to provide a clear description of the typical military blood donor. Future experimental research can now be designed with the aim of developing efficient and effective blood donor recruiting and retention campaigns. Specifically, the understanding of the demographics of the population allows targeted interventions to underrepresented groups, and theoretical research will further guide interventions that target those important motivational factors influencing blood donation.

Enamel demineralization in the form of white spot lesions (WSLs) around fixed orthodontic appliances is a persistent problem in patients with poor oral hygiene.These lesions can form rapidly within 4 weeks of bracket placement.The purpose of this in-vitro study was to investigate the fluoride recharging capability ofa commercially available orthodontic primer used to minimize the development of WSLs in patients. The three groups tested were: OpalSeal (n=20, Ultradent, South Jordan, UT), ProSeal (n=20, Reliance, Itasca, IL) and Transbond XT (control, n=10, 3M Unitek, Monrovia, CA). The samples(5mmin diameter x 1mm in thickness) weresuspended individually in vials filled with 10mL of deionized water usinga fishing line. The baseline fluoride ion release from all of the samples was measured after two weeks of changing the solution every other day. The samples were then randomly divided into two groups, toothbrush or gel. The samples in the toothbrush group were brushed for one minute every day for 7d, with fluoride containing toothpaste (Colgate-Palmolive Company, New York, NY) and placed in a new solution after each brushing. After 7d of brushing the fluoride ion release was measured. The samples in the gel group were immersed in 10mL of acidulated phosphoric fluoride gel (APF) for one minute, following manufacturer’s instructions, and then placed in a new vial with 10mL of deionized water. At the end of 24hrs fluoride ion release measurementswere made and the samples were placed individually in a new solution. The solution was changed weekly in the gel group over six weeks to simulate the typical length of time between two orthodontic appointments. A final fluoride ion release measurement was taken of all the discs in the gel group 6 weeks after the fluoride gel treatment. The results of repeated-measures analysis indicated that there were no significant differences between the groups at baseline and after 7d of toothbrushing time points. Opal Seal exhibited a significant increase in fluoride uptake (1.0ppm) after 24hrs of fluoride gel exposure but these levels gradually decreasedover 6 weeks (0.04ppm). Pro Seal and Transbond showed no significant fluoride release after the gel or toothpaste applications. The fluoride-containing primer, Opal Seal, had the ability to be recharged with fluoride ions from APF gel. However, the amount of fluoride released from recharged discs decreased gradually over a 6 weeks of time.

Epigenetics is the study of heritable gene expression due to alterations in the DNA structure other than the underlying DNA sequence. DNA methylation is one of the three types of epigenetic modifications found in the eukaryotic system. It involves the incorporation of a methyl group at the 5-position of cytosine residues in the DNA. DNA methylation is associated with several notorious disorders and diseases including Fragile X Syndrome, neurodegenerative disease (Parkinson’s, Alzhiemer, etc), diabetes and cancer. Cytosine methylation of mitochondrial DNA (mtDNA) was first demonstrated several decades ago but the mechanism of generating cytosine modification and its functional importance remain elusive. Our laboratory recently demonstrated that the enzyme involved in cytosine modification of mtDNA is a novel mitochondrial isoform of DNA Methyltransferase 1, mtDNMT1. This protein is encoded in the nucleus and targeted to the mitochondria via a N-terminal targeting sequence. Bioinformatic analysis of the DNMT1 coding sequence showed a consensus NRF1 binding site that coincidently overlaps a p53 binding site within the promoter region, previously shown by this group to repress DNMT1 expression. Previous studies in the Taylor laboratory showed that mtDNMT protein expression was regulated by the transcription factor NRF1 as well as its coactivator PGC1α. PGC1α and NRF1 stimulate a large body of genes that are involved in mitochondrial biogenesis and cellular respiration in response to environmental stress. Considering the previous findings in our laboratory regarding mtDNMT1 regulation and the importance of PGC1α and NRF1 in oxidative homeostasis, we asked whether there is a mitochondrial epigenetic component in the cell’s response to cellular stress and whether up-regulation of mtDNMT1 might be part of the general response to this stress. To investigate the relationship between mtDNA methylation and oxidative homeostasis we examined the regulation of mtDNMT1 by transcription factors that respond to oxidative stress. Conditions that induced oxidative stress were applied to HCT 116 and SH-SY5Y cell lines and the protein expression of DNMT1 was observed. Ethanol and hypoxia- induced oxidative stress were observed to increase to protein level of mtDNMT1 while total DNMT1 level either remained constant or decreased. The protein level of PGC1α and NRF1 remained low in HCT 116 cells exposed to hypoxic stress, despite elevated mtDNMT1 protein level. ChIP analysis of HCT 116 cells exposed to hypoxic stress demonstrated that NRF1 and PGC1α are not regulating the transcription of DNMT1i in the mitochondria. However, we observed that p53 dissociated from the DNMT1 promoter upon hypoxic stress, indicating that the up-regulation of mtDNMT1 is through the relief of p53 suppression. The findings of this investigation proved that mtDNMT1 is receptive to oxidative stress through the regulation by p53 and suggested that mitochondrial epigenetics may be playing an integral role in the cellular stress response toward hypoxia.

Neisseria gonorrhoeae requires iron for survival and establishment of infection in the human host. Pathogenic Neisseriae have evolved a repertoire of high-affinity iron acquisition systems to facilitate iron uptake in the human host. This requires specific outer membrane receptors and energy-harnessing cytoplasmic membrane proteins. The transferrin receptor proteins of Neisseria gonorrhoeae are necessary for iron uptake from transferrin in the host. The iron uptake system consists of two transferrin binding proteins, (Tbp) A and B. TbpA is an integral outer membrane, TonB-dependent transporter that forms the pore for iron internalization. TbpB is a surface exposed lipoprotein that makes the iron internalization process more efficient. TbpA is proposed to consist of two distinct domains: a β barrel and an N-terminal plug domain. Previous studies have shown that the EIEYE sequence in the TbpA plug domain plays an important role in iron internalization. We undertook a collaborative project to test the hypothesis that the conserved EIEYE sequence in the wild-type TbpA plug binds Fe3+ during the outer membrane iron transport process. CD spectra analysis and fluorescence emission titration spectra of purified recombinant wildtype and mutated plug proteins revealed that Fe3+ is sequestered by the wildtype TbpA plug protein, unlike the mutated plug protein. Modeling data with the wild-type plug predicts the EIEYE sequence is part of a flexible loop structure and acts as an Fe3+ binding site. Characterization of the Tbps constituting the gonococcal receptor is important to understanding how the gonococcus survives within its host. TbpA and TbpB act together to acquire iron from human transferrin. We hypothesize that the presence of TbpA impacts the exposure or conformation of TbpB. In this study, we have utilized photoactivable cross-linkers to assess the effect of TbpA on TbpB in live gonococcal cells and studied it in presence of ligand and TonB derived energy. We employed insertion mutants, in which TbpA and TbpB contained the hemagglutinin (HA) epitope tag, to probe for impact of TbpA on TbpB. Our results demonstrate that photo-cross-linking altered TbpB size and migration and was dependent on the presence of TbpA. HA epitope insertion mutants in surface exposed loops of TbpA and TbpB did not impact the mobility of cross-linked TbpB. Addition of human transferrin to the de-energized mutant caused a change in TbpB migration after cross-linking. This result indicates that when ligand is bound tightly and irreversibly to de-energized TbpA, the surface accessibility and perhaps conformation of TbpB is altered and TbpA does not interact with TbpB. Our findings were confirmed with recent structural studies of TbpA-TbpB-ligand triple complex, which illustrate that Tbps bind ligand through unique, non-overlapping binding sites such that TbpA and TbpB do not interact. TbpB is not an essential member of transferrin-iron acquisition pathway. It is surface exposed and tethered to the outer membrane by a lipid moiety. The role of TbpB has not been clearly outlined in the transferrin iron acquisition system. The last objective of this study was to look at the significance of two specific conserved regions of TbpB and its importance in transferrin iron utilization and TbpA-TbpB interaction. Using site-directed mutagenesis we created two mutants, in the first mutant the conserved lipobox of TbpB was replaced with a signal I peptidase cleavage site, and the second mutant contained a deletion of the conserved poly glycine residue stretch, immediately downstream of the lipobox. Our results indicate that lipobox is required for lipidation of TbpB, both the mutants were impaired for transferrin-iron utilization, and neither of the mutations altered TbpA-TbpB interaction. Overall, these studies help elucidate the functional importance of the specific regions in TbpA and TbpB in Neisseria gonorrhoeae, thereby adding to our understanding of the process of iron acquisition through the transferrin binding proteins.