Background: Pressure ulcers (PrUs) have a significant impact on health system expenditure and patient’s quality of life. It is a global problem. Many studies were undertaken in regard to PrU prevention and management. In Oman, no studies have been conducted to investigate nurses’ knowledge on prevention and management of PrUs. The purpose of this descriptive sequential explanatory mixed-method study was to explore the nurses’ level of knowledge in relation to prevention and management of PrUs in Oman. Methods: A mixed method design was used and the study was conducted over two Phases. In Phase I, a questionnaire was developed to explore nurses’ knowledge on PrU, policy, and resources. The main section of the questionnaire was the Pieper-Zulkowski Pressure Ulcer knowledge test (PZ-PUKT) which tests the knowledge on PrU. Another two sections were developed including questions about wound policy and resources available for PrU prevention and management in Oman. The questionnaire was distributed to nurses who were working in surgical, medical, orthopaedic, CCU, and ICU wards/units in seven hospitals. In Phase II study, semi-structured qualitative interviews were conducted with 16 of the questionnaire respondents. Interviews took approximately 30 minutes, were recorded and transcribed verbatim. Qualitative data were analysed using the Knowledge, Attitudes and Practice (KAP) model as the a priori framework. Results: In Phase I, 478 questionnaires were analysed. The knowledge test results showed the overall mean percent score for correctly answered questions was 51% suggesting a low level of knowledge. There was a significant relationship between nurses’ knowledge and age (P=0.001) and between knowledge and years of experience (P=0.001) with knowledge increasing with age and years of experience. In Phase II, four themes were identified from the interviews: knowledge, attitude, and practice (framework themes) and perception of role. Findings indicated positive and negative attitudes towards the care of PrUs. Some nurses stated feeling rewarded when they see wounds improving while others said they could not work with patients independently because they lacked the knowledge and the skills needed. There was variation in the management of PrU between hospitals. Both studies indicated that the wound management policy did not include enough information to guide nurses. Conclusion: Overall the nurses’ level of knowledge on PrU was relatively low. Most nurses were not familiar with wound management policy or different PrU prevention and management strategies. Nurses are aware of the risk of PrUs and try their best to manage them with the available resources however more training is required.

Transplantation of genetically engineered neural stem cells (NSCs) into sites of central nervous system (CNS) disease/injury is a promising strategy to promote repair of damaged tissue. However, translating this strategy into the clinic requires several challenges to be overcome including facilitating ‘combinatorial therapy’ (achieving multiple therapeutic goals – essential in CNS injury/disease). Nanotechnologies are emerging as multifunctional platforms capable of meeting this requirement. For example, magnetic particles (MPs) and implantable hydrogels offer several biomedical advantages for transplant populations, including: safe genetic manipulation; non-invasive cell tracking, via MRI; and safe and efficient accumulation of cells at sites of injury. However, the use of these nanotechnologies remains to be explored in detail for NSC transplantation therapies. In this thesis, it is shown that MPs can mediate gene delivery to NSCs grown as neurospheres and monolayers with the most efficient transfection efficiencies achieved using oscillating magnetofection protocols (9.4% and 32.2% respectively). In both culture systems, developed protocols had no effect on key regenerative properties of NSCs such as cell viability, proliferation, stemness and differentiation. Further, ‘magnetofected’ monolayer NSCs were shown to have survived and differentiated in a cerebellum slice model acting as host tissue, indicating safety of the procedures. It was also shown that assessing procedural safety and extent of transfection of magnetofection protocols may be feasible by employing mass spectrometry and proteomics analysis. It was also found that tailored enhancement of particle magnetite content offers a means to efficiently label NSCs, up to a maximum of 95.8%. Labelling procedures had no effect on cell viability, proliferation, stemness or differentiation. In addition, labelled cells could survive and differentiate in a slice model of spinal cord injury indicating safety of the labelling procedures. Functional labelling was also demonstrated by magnetic capture of labelled cells in an in vitro flow system. Hydrogels offer major advantages for delivery of transplant populations into injury sites. Here it was shown that an intraconstruct genetic engineering approach was feasible for NSCs cultured with a clinically translatable, collagen hydrogel system. Magnetofection protocols safely increased MP mediated transfection of NSCs grown in ‘2-D’ and ‘3-D’ hydrogel cultures.

More than 90 serotypes of Streptococcus pneumoniae are responsible for causing a number of invasive and non-invasive diseases in humans. To combat this pathogen there are two types of vaccines available namely pneumococcal polysaccharide vaccine (PPV) and pneumococcal conjugate vaccine (PCV). Although these vaccines have reduced the incidence and prevalence of pneumococcal diseases yet these vaccines are unable to curb the disease and have some potential drawbacks. The problems associated with PPV is inability to mount appropriate immune response in young children (<2 years) of age and with PCV, disadvantages are serotype replacement and affordability. These disadvantages have shifted the focus of attention on developing vaccines that can provide serotype independent protection and is cost effective and can be used in developing and under-developing countries as well. Pneumolysin (PLY) is one of the members of cholesterol dependent cytolysin (CDC) family and is one of the key virulence factors of the bacterium. PLY has a wide variety of functions but the two most important functions are the ability to lyse the host cell by forming pores and activating the complement pathway. Studies have shown recombinant PLY to be a good vaccine candidate; however PLY is toxic and cannot be used as a vaccine. A number of toxoid versions have been made targeting residues responsible for toxin’s lytic activity and tested in-vivo models. ∆6 PLY is a toxoid made by deleting two amino acid residues and seems to be a potential candidate to be included in next generation pneumococcal vaccines. ∆6 PLY is immunogenic and act as an adjuvant as well, however ∆6 PLY causes the aggregation of red blood cells and therefore cannot be used in the vaccine. The present study is carried out to identify the residue/s responsible for causing this agglutination behaviour. A number of molecular biology techniques were used in this study for making mutants in different backgrounds and were tested for their lytic and agglutination activity. The study identified the single residue in domain 4 causing aggregation of the red blood cells. The agglutination negative mutant was then tested at different concentrations through a series of experiments including hemagglutination assays, fluorescence microscopy and fluorescence activated cell sorter (FACS). The study also highlighted some important regions of the toxins responsible for maintaining structure of the toxin. This new toxoid seems to have the potential to be used in future pneumococcal vaccines alone, in combination with other pneumococcal proteins and/or polysaccharides.

Occupational Therapists (OTs) who recommend housing adaptations for people with disabilities, funded through public finance, must satisfy professional codes of practice and the employing local authority requirement to allocate finite resources effectively and fairly. At the same time they must also meet service user expectations. Ethical reasoning will be required to balance these demands whilst practising to a personally acceptable professional standard. This study investigates how OTs understand themselves to develop a sense of fairness and how they use their community of practice in developing professional ethical practice. This was a 2 part methodology. Firstly, OTs from 2 community services were invited to participate in small discussion groups. 3 group sessions, of different sizes ranging from 2-6 participants and duration of 2-3 hours, were recorded in which OTs discussed cases which posed ethical challenges with respect to fairness. All participants were female. Secondly, 4 individual interviews with volunteers from the groups were recorded to collect OT narratives of personal ethical development. Transcripts were analysed using a literary-critical approach focussing for transcripts of group sessions on dialogue in community of practice and ethical approaches used; focussing for interview transcripts on the process of ethical development. OTs were shown developing professional practice dialogically within their own community of practice groups. This finding confirms the importance for professional development of encouraging opportunities for dialogical interaction between OTs. Practical reasoning about justice as theorised by Sen (2009) better characterised OT ethical reasoning practices than biomedical-ethical approach applying universal, abstract ethical principles. OT narratives of ethical development fitted the Aristotelian model of growth in virtue as a whole, across both professional and personal aspects of life. Empathy was tentatively categorised as a virtue rather than a technical skill in this context. Empathy contributed to OT clinical reasoning processes as well as ethical reasoning.

Nowadays, the long-haul optical fiber communication system is one of the important way to convey the information, and there is strong competition of research in the optical long-haul transmission to achieve high channel bit rates and large transmission capacity. Therefore, it is important to study a technology to improve the performance of such system. The return-to-zero differential phase-shift keying (RZ-DPSK) and the return-to-zero differential quadrature phase-shift keying (RZ-DQPSK) have received renewed attention recently for the long-haul transmission systems, because they can improve the transmission performance of the long-haul system. Furthermore, the design of the dispersion map becomes significantly different from that of the conventional system using the intensity modulation direct detection (IM-DD) scheme. Besides, the RZ-DQPSK can transmit two bits per symbol, so it has twice the spectral efficiency of the RZ-DPSK. This study is focusing on the difference of the transmission performance of the long-haul RZ-DQPSK system due to the dispersion map using the numerical simulation.

BACKGROUND: Aggression can have a wide range of damaging consequences for both perpetrators and victims. Theoretical and empirical studies into problems of aggression increasingly show the importance of social and cognitive factors in aggressive behaviour. Such research has commonly been approached through the framework of the Social-Information Processing (SIP) model. SIP explains social behaviours by the sequence of cognitive processes that occur between encountering a social stimulus and enacting a response to it. Crucially, it is apparent that particular processing styles, such as the way in which people interpret others’ behaviour, play important roles in aggression. However, while SIP has long been used to explain aggression in the non-disabled population, it is only in more recent years that this approach has been applied to people with intellectual disabilities (IDs). This is important because a significant minority of people with IDs demonstrate frequent aggressive behaviour. Although several studies have already indicated that particular cognitive processing tendencies and aptitudes contribute to aggression in adults with ID, no research has considered younger people in the transition to adulthood. To this end, the present thesis sought to investigate the possible influences of certain psychosocial factors on this group of young people with mild to moderate IDs. OBJECTIVES: To identify which specific factors to investigate, a systematic review was conducted of existing research into SIP and aggression with people who have IDs. On the basis of these findings, the thesis examined 1) the social interactions that typically elicit anger, 2) experiences of parental aggression 3) ability to discern affect from dynamic social cues and 4) beliefs about the consequences of aggressive and submissive behaviour. With the review also stressing the importance of examining aggression at specific developmental stages, the studies focused on individuals in the transition from adolescence to adulthood (between 16 and 20 years). Although this stage is thought to be important in the development of cognitive factors associated with aggression, there is little or no research in this area with young adults with IDs. METHODS: The thesis comprised four distinct research studies. Each adopted a group-comparison design, comparing aggressive and non-aggressive young people with IDs. To evaluate the extent to which findings were specific to people with IDs, additional comparisons were conducted between aggressive and non-aggressive individuals without IDs. For Study 1, 26 young adults with IDs and 20 non-disabled young adults completed a semi-structured interview about a recent experience of interpersonal conflict. Participants were asked to describe their beliefs and feelings about the event and their subsequent response. Studies 2, 3 and 4 used data from a second phase of data collection involving 46 young people with and 48 people without IDs. Study 2 used a task in which participants were asked to rank different types of social conflict in order of provocativeness. The author developed these scenarios to reflect the experiences of conflict reported by participants in Study 1. Participants also indicated how recently they had encountered each type of scenario. Study 3 used motion-capture stimuli of people walking in different emotional states to examine whether groups differed in how they encode dynamic social cues. Study 4 used provocative vignettes to examine whether aggressive young people with IDs expect different outcomes from aggressive and submissive responses to such scenarios. RESULTS: Study 1 found that participants with IDs were more likely to encounter conflict with strangers or peers outside their friendship group. They were also more likely to describe incidents of aggression and to characterise people with whom they were in conflict globally as “bad” and to perceive their actions as being personally directed at them. Study 2 did not suggest that experiences of being victimised by peers were more common for people with IDs, but did show that aggressive individuals were more likely to encounter incidents of physical aggression from peers. Parental conflict was the most recently encountered, but was perceived to be the least provocative form of conflict for all groups. In Study 3, no group differences were found in accuracy or response tendencies for the emotion recognition task. Aggressive and non-aggressive participants with IDs in Study 4 did not predict different outcomes form aggression and submission. However, the aggressive participants without IDs predicted more positive outcomes from aggression and more negative outcomes for submission. While aggressive participants with IDs were more likely to give aggressive responses, they were just as likely as the non-aggressive group to respond actively (assertively or aggressively) rather than passively. CONCLUSION: The findings of this thesis, viewed from the perspective of the SIP model, suggest that there are key cognitive and contextual differences between individuals who show frequent aggression, both with and without IDs. Although, somewhat surprisingly, emotion recognition skills did not appear to be associated with a tendency toward aggressive behaviour, the non-ID aggressive and non-aggressive groups differed in their anticipated outcomes for aggressive and submissive behaviour. The context in which conflict occurred also appeared to differ between those young people with and without IDs. However, the absence of some predicted findings from these studies may be related to methodological shortcomings; these possible limitations are considered, and directions for future work are suggested. Applications for clinical practice and policy are also discussed and recommendations for future research are given.

The process of parturition resulting in the delivery of a newborn is a fundamental event ensuring survival of the species. In humans, the main clinical problems of parturition include activation of the process too early or too late resulting in the delivery of pre-term and post-term infants, both with their own implications for future health for the mother and baby. Additionally, where parturition systems are not activated correctly, dysfunctional labour with the resulting need for caesarean delivery (CS), in addition to atonic post-partum haemorrhage can also ensue. Overall, in the UK up to 40% of pregnancies are affected by one of these problems. However, the exact processes involved in the initiation and maintenance of parturition in the human are not fully understood. With such an important event, influences are most likely to be multi-factorial, with hormonal, mechanical, inflammatory, biochemical and maternal environmental factors playing a part. The aims of this thesis were to investigate influences on parturition in human pregnancy. Firstly, the myometrial transcriptional effects of long term exposure to the uterotonic oxytocin (OT) were examined. Further investigation of the myometrial and maternal peripheral response to uterine contractions in-vitro and in-vivo was also made with particular reference to the role of inflammation and myocyte damage. Additionally, the influence of maternal factors, particularly obesity, on the myometrial in-vitro contractile function and response to OT was studied. Initially, 150 gene arrays were produced using the Illumina platform. The samples were derived from myometrium taken at pre-labour CS which subsequently underwent functional contractility experiments in an organ bath. Five drug environments were studied, namely OT, acetic acid (OT vehicle), ML7(a tocolytic acting via inhibition of myosin light chain kinase), ML7 & OT and finally DMSO (ML7 vehicle). Additionally, five time-points of 0, 1, 2, 4, and 6 hours after drug addition were used, resulting in 5 samples for each drug and time combination. The results indicated that despite a clear enhancement of myometrial contractile activity by OT, this functional response does not appear to be mediated by cellular transcription. However, there was a clear contraction and time dependent transcriptional wave, with overrepresentation of genes associated with inflammation and cellular damage/apoptosis, and down-regulation of pathways concerning cellular metabolism. These findings were confirmed by QPCR on further myometrial samples undergoing additional in-vitro functional studies. In addition to the temporal and contractile association with the inflammatory response, our data suggest inflammation occurs in response to myocyte cellular damage regardless of mode of damage e.g. contractile or chemically induced. This was demonstrated by inflammatory upregulation in myometrium exposed to the tocolytic agents nifedipine and ritodrine, which is not seen in response to ML7. Additionally, the myometrial inflammatory response was enhanced by the infective agent LPS. However, contrary to other proposals, the enhanced inflammatory response of the myometrium did not alter or promote the in-vitro contractile ability of the myometrium or its response to OT. This myometrial transcriptional data therefore suggests that the inflammatory response of labour is associated with contraction, chemical or infection induced myometrial cellular damage, but would not be considered necessary for a contractile response. Our in-vivo study of peripheral changes in the maternal circulation again supported our in-vitro myometrial data. Data showed that the effect of pregnancy at term was limited to increased white cell count driven by a neutrophilia, with no suggestion of leukocyte priming prior to labour. Additionally, term pregnancy is associated with an increase in CRP, an increase in GCSF (corresponding with the neutrophilia) in addition to suppression of the chemokines CCL11 and CCL22. Subsequently, we found that repeated blood samples taken at 2 hourly intervals during term labour induced dramatic changes in inflammatory cells and inflammatory mediators in the maternal circulation. Importantly, these changes occur in a co-ordinated time and contraction dependent manner, with the degree of inflammation associated with the length of time in labour and the degree of myocyte damage as measured by circulating CK and Mb. Our study of the influence of maternal factors on myometrial contractile ability and response to OT examined in-vitro myometrial contractility of 609 myometrial strips from 85 women. We demonstrated that maternal obesity does not impair spontaneous or OT induced myometrial contractions in-vitro. Furthermore, maternal age, ethnicity, parity, previous caesarean delivery,gestation at delivery and birthweight do not influence in-vitro myometrial spontaneous or OT induced contractile activity. This therefore suggests that the observed implication of these maternal and infant factors on parturition in-vivo (high rates of induction of labour, high rates of intrapartum caesarean delivery and post partum haemorrhage) cannot be explained by an effect on myometrial contraction per se. This therefore merits further investigation as to alternative mechanisms to ultimately promote and effective, uncomplicated and safe labour and vaginal delivery for at risk mothers. In summary, this thesis provides evidence that the myometrial contractions of human labour, whether spontaneous or OT induced are capable of inducing a temporal wave of transcriptional changes associated with the processes of inflammation, cellular damage/apoptosis with inhibition of cellular metabolic processes. In addition, maternal peripheral circulating factors mirror the myometrial transcriptional changes. These changes are highly comparable with those seen in response to exercising skeletal muscle, and in this model have been shown to play an important role in muscle repair and remodelling after exercise. Therefore, we would suggest that the inflammatory reaction typically associated with human labour occurs as a non-specific response to contraction induced cellular damage and may play a role in postpartum repair and remodelling of the uterus.

Background: There is a high cellular turnover in the haematopoietic system which necessitates that new cells are continuously produced to replace old and senescent ones. The haematopoietic stem cells and its progenitors meet this requirement. In adults, haematopoietic stem and progenitor cells (HSPCs) generally occupy a unique microenvironment in the bone marrow called the niche. However, recent findings have shown that approximately 0.06% of HSPCs circulate between bone marrow and periphery in steady state. Evidence shows that HSPC express cytokine receptors and pathogen recognition receptors [e.g. Toll-like receptors (TLRs)], which suggest that HSPCs may directly respond to inflammation and infection. The capacity of HSPCs to directly respond to infection is demonstrated in models of bacterial infection using LPS injection that significantly increases the number of circulating HSPCs. However, the underlying mechanism is not clearly understood. As chemokines orchestrate in vivo cellular migration, it was hypothesised that HSPC inducibly express inflammatory chemokine receptors that enable them to respond to circulating chemokines during infection and inflammation. Methods and results: Here, the impact of systemic or peripheral inflammation on HSPC of mice was investigated using LPS injection and topical imiquimod cream/TPA treatment respectively. Using haematopoietic progenitor colony-forming assays, RT-QPCR on isolated progenitors, gene-knockout mice, flow cytometric analysis and in vivo antibody-mediated neutralisation experiments, data are provided showing that HSPC inducibly express chemokine receptors in response to inflammation. Critically, the topical imiquimod inflammation model required functional chemokine receptor 2 (Ccr2) for HSPC mobilisation in contrast to both systemic LPS and topical TPA models, which were Ccr2-independent. Furthermore, dermal inflammation was necessary for imiquimod-mediated HSPC mobilization, as subcutaneously administered imiquimod did not result in significant HSPC mobilization. Conclusion: The data suggest that, in addition to the established CXCR4-CXCL12 axis that regulates homeostatic HSPC trafficking, the inflammatory chemokine-chemokine receptor axes may also be crucial in modulating HSPC functions during infection and inflammation.

D6 is an atypical chemokine receptor related to CCR1-5 that binds to many inflammatory CC chemokines. Experiments using transfected cell lines have shown that upon binding to a chemokine ligand D6 does not trigger cellular signalling pathways, but rather acts to scavenge the bound ligand. It achieves this by constitutively travelling to and from the cell surface via early and recycling endosomes, internalising chemokines bound when it is at the cell surface. Over time, D6 removes a large amount of ligands from the extracellular compartment. In vivo, this scavenging activity is thought to regulate the level of CC chemokines, and thus controls inflammation locally and systemically. Lack of D6 has been shown to result in elevated amounts of bioavailable chemokines, and is associated with over exuberant inflammatory responses. In human, D6 mRNA and protein is highly expressed in trophoblast-derived gestational tissues. The expression of D6 mRNA in the placenta is by far the highest, compared to other solid tissues being studied. The importance of D6 in protecting the offspring has been demonstrated in animals. In pigs, a defect in D6 expression was discovered in placental attachment sites in endometrium from arresting fetuses. In mice, lack of D6 results in an increase in fetal loss after challenge with lipopolysaccharide (LPS) or antiphospolipid autoantibodies (aPL), and an increase in the number of abnormal pups when mouse embryos are transferred into fully allogeneic pseudo-pregnant female recipients. In view of these results suggesting a critical role for D6 in placental mediated complications, the expression and molecular function of D6 in primary human trophoblast cells were studied, as to date in vitro human studies have utilised the choriocarcinoma cell line BeWo or immortalised cell lines engineered to over-express exogenous D6. Secondly the impact of D6 deficiency on placental structure, chemokine expression and leukocyte abundance in mice was examined. Chapter 3 presents the results of experiments on primary human trophoblasts. Protocols for routine primary trophoblast isolation, purification and culture from fresh term placentas were optimised in our laboratory. D6 mRNA was detected in these primary cells. Using Western blotting, immunofluorescence and flow cytometry, D6 was shown to be present predominantly in the intracellular vesicles of the cells. Competition chemokine uptake assays, analysed by flow cytometry, showed that CCL2 was internalised by trophoblasts using D6. Competitive chemokine scavenging assays, analysed by quantitative Western blot, confirmed that D6 was functioning as a chemokine scavenger on primary human trophoblasts and that it progressively removed substantial quantities of chemokine from medium bathing the cells. This is the first set of experiments that confirms D6 is present, and functioning as a chemokine scavenger in primary human cells. Chapter 4 contains the results from the mouse experiments. Even in an unchallenged environment it was shown that, on the DBA-1 genetic background, D6 deficiency in the mother and pups leads to higher rates of stillbirth and neonatal deaths, resulting in a reduction in the number of pups weaned per litter than their WT counterparts. By gestational age E14, pup weight was significantly smaller in the D6 KO mice. Using stereological techniques, the placenta of the D6 KO mice at this gestation was found to have a smaller labyrinthine zone. The volume of the labyrinthine zone was positively correlated with pup/placenta ratio. These phenotypes could be due to a maternal or fetal effect of D6 deficiency. To ascertain the answer to this question, the experiment at E14 was extended by breeding DBA-1 females heterozygous for the deleted D6 allele (D6 HET) with D6 deficient (D6 KO) males. In this model the phenotypes of D6 KO pups and placentas could be compared with their D6 HET siblings that developed in a mother expressing some D6 (i.e. D6 HET). Although there were no differences in pup weight, placental weight and pup/placenta ratio between these two groups, stereology revealed a decrease in labyrinthine zone volume fraction in the D6 KO placentas in comparison to their D6 HET siblings. The observed fetal compromise and placental defect at E14 was not apparent at the later gestational age of E18. Luminex multiplex protein assay showed an elevated level of circulating chemokine CCL2 in the serum of D6 KO pregnant mice in comparison to their WT counterpart, so loss of chemokine regulation could be responsible for the defects observed in D6 deficient placentas. In summary, D6 deficiency results in an increase in perinatal death, a fundamental defect in placental formation (reduced labyrinthine zone) and dysregulation of circulating chemokine levels. Chapter 5 discusses the mechanisms of D6 in regulating placental formation and reproductive outcome and the novel insights that this work provided into placental D6 function. It also describes the design of future experiments to reveal the precise role of D6 in chemokine regulation and cell signalling in reproductive immunology, and discusses how D6 might contribute to pregnancy outcome in humans.

Background: In the past 60 years, the median age of the entire world population has increased and ageing and chronic diseases are now the main medical concerns in the developed world. The identification of early signs of disease pathogenesis is vital for prevention and targeting populations at risk in order to reduce morbidity and mortality. Glycation is well-established as an index of control, or otherwise, and a predictor of end-organ damage, for people with type 2 diabetes. At the beginning of the work for this thesis (2010), evidence was beginning to be raised to suggest that since glycation levels vary considerably, in normoglycaemic, non-diabetic individuals, glycation cannot be solely related to glucose levels and early reports allowed for speculations about a relationship between early glycation and oxidative stress. The aim of this thesis was to establish the relationship between early glycation and oxidative stress in normoglycaemia using techniques from the full breadth of Human Nutrition Research. Methods: In study 1, existing epidemiological data were used to identify relationships between proxies for redox status and early glycation in non-diabetic individuals. One-way ANOVA, Chi-squared and multivariate linear regressions, adjusted for all known confounders were used to explore associations of HbA1c with self-reported smoking status and fruit & vegetables consumptions in the Scottish Health Surveys 2003-2010, among individuals without known diabetes and HbA1c<6.5%. In studies 2 and 3, the associations from epidemiology were explored in mechanistic laboratory studies with high physiological relevance (using physiological concentrations and conditions) to better characterise the effect of oxidative stress and antioxidants on early glycation. In study 2, bovine serum albumin (BSA), reduced BSA (mercaptalbumin) (both 40g/L), and human plasma were incubated with glucose concentrations 0-30 mM for 4 weeks at 37oC. All were tested pre-oxidized for 8 hours prior to glycation with 10nM H202, or continuously exposed to 10nM H2O2 throughout the incubation period. Fructosamine was measured (nitroblue tetrazolium method) at two and four weeks. In study 3, Bovine Serum Albumin (BSA) was pre-treated prior to in vitro glycation: either no treatment (native), pre-oxidised (incubated with 10nM H2O2, for 8 hours) or incubated with a mixture of phenolic acids at physiologically relevant concentrations, for 8 hours). In-vitro glycation was carried out in presence of i) glucose only (0, 5 or 10mM), ii) glucose (0, 5 or 10mM) plus H2O2 (10nM), or iii) glucose (0, 5 or 10mM) plus phenolic acids (10-160nM). Fructosamine was again measured using the nitroblue tetrazolium method. Prior to the experimental study we carried out a systematic literature review of dietary interventions reporting plasma concentrations polyphenol metabolites, to inform the design of a physiologically relevant in-vitro study. In study 4, clinical trial data and biological samples were analysed from a randomised controlled dietary advice trial in obese pregnant women, a group at risk from higher glycation and oxidative stress. Samples and data from the UPBEAT study trial (n=117) were analysed. Plasma fructosamine, plasma sRAGE, urinary Ferric Reducing Ability of Plasma (FRAP), urinary Total Phenol (TP) and urinary Advanced Oxidised Protein Products (AOPPs) were measured at 16-18+6 and 27-28+6 weeks gestation. Dietary recalls were used to calculate fruit and vegetable and polyphenol intake at the same timepoints. Data were analysed to identify associations between dietary variables and biochemical markers, as well as their relationships with diagnosis of complications. Associations between maternal variables and neonatal anthropometry were also investigated. Results: In study 1, HbA1c was higher in smokers by 0.25 SDs (0.08%), and 0.38 SDs higher (0.14%) in heavy smokers (>20cigarettes/day) than non-smokers (p<0.001 both). Smokers were twice as likely to have HbA1c in the ‘pre-diabetic’ range (5.7-6.4%) (p<0.001, adj.model). Pre-diabetes and low grade inflammation did not affect the associations. For every extra 80g vegetable portion consumed, HbA1c was 0.03 SDs (0.01%) lower (p=0.02), but fruit consumption did not impact on HbA1c, within the low range of consumptions in this population. In study 2, oxidized BSA (both pre-oxidised and continuously exposed to H2O2) was more readily glycated than native BSA at all glucose concentrations (p=0.03). Moreover, only oxidized BSA was glycated at physiological glucose concentration (5mM) compared to glucose-free control (glycation increased by 35% compared to native albumin p<0.05). Both 5mM and 10mM glucose led to higher glycation when mercaptalbumin was oxidised than un-oxidised (p<0.05). Fructosamine concentration in human plasma was also significantly higher when oxidized and exposed to 5mM glucose, compared to non-oxidised plasma (p=0.03). The interaction between glucose concentration and oxidation was found to be significant in all protein models (p<0.05). In study 3, the presence of six phenolic acids with BSA during in-vitro glycation did not lower fructosamine formation. However, when BSA was pre-incubated with phenolic acids, significantly lower concentration of fructosamine was detected under glycoxidative conditions (glucose 5 or 10mM plus H2O2 10nM) (p<0.001 vs. native BSA). In study 4, women in the lowest quartile of total polyphenol intake had 8% greater fructosamine levels compared to those in the top quartile. Total polyphenol intake was negatively correlated with sRAGE levels. Diagnosis of severe preeclampsia was associated with elevated AOPPs. Maternal polyphenol intake was positively correlated with birth weight, while maternal glycoxidation showed the opposite relationship. Conclusions: Study 1 added evidence to relate smoking (an oxidative stress proxy) to protein glycation in normoglycaemic subjects. This association has implications for individuals exposed to ROS and for epidemiological interpretation of HbA1c and its clinical usefulness. Study 2 offered a mechanistic background to the previously shown epidemiological association. This study demonstrated for the first time albumin glycation in-vitro, using physiological concentrations of albumin, glucose and hydrogen peroxide. These results identified low-grade oxidative stress as a key element early in the glycation process, especially in glucose concentrations relative to normoglycaemia. Furthering those findings, study 3 showed that protein-phenolic acid interactions are important regulators of protein glycation. Together those studies highlighted that protein pre-treatment, either with oxidants or phenolic acids, is an important regulator of subsequent glycation in a physiologically relevant system. An important outcome of those studies is that high quality in-vitro studies under conditions closer to physiology are feasible and should be employed more frequently. Finally, study 4 demonstrated an association between polyphenol intake and glycation during pregnancy, with an impact on neonatal outcome measures. Maternal glycoxidation is a promising marker of pre-eclampsia and neonatal anthropometry and could be modulated by maternal lifestyle and dietary habits. Overall, the results of this thesis implicate that drivers of redox status have the capacity to modulate protein glycation in normoglycaemia. These results challenge the assumption that glycation levels are solely dependent on circulating glucose levels and suggest a useful application of glycation outside the field of diabetes.