Inducible nitric oxide synthase (iNOS)-induced nitric oxide (NO) and its reactive metabolite peroxynitrite (ONOO־) has been implicated as an important mediator in the pathogenesis of inflammatory bowel disease (IBD). Immune cells macrophages produce NO and ONOO־ to provide the host defence and play important roles in the regulation of inflammatory response. Several in vitro and In vivo studies have reported the role of probiotics and probiotic-released metabolites in the treatment and prevention of IBD. The study was therefore started with the hypothesis that the metabolites released by probiotics present in the formulation VSL#3 may have the effect on the pro-inflammatory mediator NO and its regulatory enzyme iNOS in the immune cells. Murine macrophage J774 cells were used in the study as these cells are readily induced by bacterial lipopolysaccharide (LPS) and produce pro-inflammatory mediators nitric oxide (NO) and prostaglandin E2 (PGE2) by the expression of enzymes iNOS and cyclooxygenase-2 (COX-2) respectively through the Nuclear Factor-κappa B (NF-B) signalling pathway. The cells were exposed to VSL#3-conditioned medium (VSL#3-CM) in the absence and presence of LPS and the inhibitors of a number of enzymes and signalling molecules. To exclude the possibility of endotoxin contamination in VSL#3-CM, experiments were also carried out in the absence and presence of an endotoxin neutralising compound polymyxin B (PmB). VSL#3-CM increased the basal NO production through the induction of iNOS on a concentration dependent manner. On the other hand, it suppressed the LPS-induced NO production but caused slight inhibition on the LPS-induced iNOS expression. PmB suppressed the LPS-induced NO production but not that caused by VSL#3-CM suggests the effects of the later were not LPS mediated. Similarly, VSL#3-CM at a higher concentration induced the basal COX-2. However, it caused slight inhibition on the LPS-induced COX-2 expression. Dexamethasone has partially inhibited the VSL#3-CM-induced NO and iNOS expression. Bisindolylmaleimide (BIM) and SB203580, inhibitor of protein kinase C (PKC) and p38 MAPK, a member of the mitogen activated protein kinases (MAPKs) family respectively, have markedly inhibited VSL#3-CM-induced iNOS expression and NO production. Slight inhibition was made by LY2904002 and CAY10470, inhibitors of Phosphatidylinocitol 3-kinase (PI3K) and NF-B respectively. Akt inhibitor XIII and MG132 the inhibitors of the protein kinase B (Akt) and proteasome showed no inhibition on VSL#3-CM-induced iNOS and NO production. A balanced expression of iNOS and NO production is necessary for the regulation of normal cellular functions. In this study, it has been shown that VSL#3 has the ability to regulate the function of iNOS and thus the over production of NO in presence of LPS. This action may explain, in part, the proposed anti-inflammatory effects of VSL#3 and thus its potential benefits in IBD. Interestingly, VSL#3 may also induce iNOS expression and NO production through the activation of PKC, p38 MAPK and PI3K pathways but only when applied alone under control conditions. Whether this is specific to macrophages or a common effect of VSL#3 is currently not clear. It would not be considered normal however to recommend VSL#3 for routine use in healthy individuals. The ability of VSL#3 to downregulate the expression and function of iNOS induced by endotoxin however, provides a good rational for its use in inflammatory disease states including IBD.

No description available.

No description available.

Diabetes is associated with abnormal nutrient metabolism particularly carbohydrates and lipids. In pregnancy, this diabetes-associated metabolic perturbation has been shown to alter, qualitatively and quantitatively. the uterine lipid environment and consequently the fatty acids delivered to the developing foetus. Perhaps not surprisingly. maternal consumption of a high saturated fat diet during pregnancy compounds the problem. The primary aims of the studies reported in this thesis were to investigate the effect of: (a) matemal diabetes on placental fatty acid composition in human subjects, and (b) matemal diabetes and a high saturated fat intake on tissue composition of the offspring in laboratory animal model. Therefore. the key objectives of this thesis were to: (1) . analyse human placental fatty acids from healthy pregnancies at two times in gestation. early (8-14 wk) and term (38- 41wk). (2) compare placental fatty acids from pregnant women with gestational diabetes mellitus (GDM) and women who delivered preterm with the healthy term placenta, and (3) investigate the effects of maternal diabetes and/or a high fat diet during pregnancy on the fatty acid composition of the vascular tissues, particularly arachidonic (AA) and docosahexaenoic (DHA) acids of the offspring. The main outcomes of these investigations were that: (1) AA was incorporated in high proportions (17.2-35.9%) in most phospholipids. At term. AA was reduced in choline (CPG). ethanolamine (EPG) and inositol (IPG) phosphoglycerides while di-hommo-ylinolenic acid (DGLA) was markedly increased in CPG, EPG, IPG and serine (SPG) phosphoglycerides compared with early in gestation (8-14wk) placental membranes, (2) the GDM placenta had elevated AA and DHA in CPG and EPG while the preterm placenta had reduced DHA in CPG and SPG and AA in SPG, and (3) maternal diabetes reduced AA and arachidonic-to linoleic acid ratio (AAlLA) in the offspring. Similarly, feeding dams to a 32.9%-fat or 20%-fat (mainly lard) diet reduced DHA in liver and heart CPG and EPG or AA and DHA in aorta CPG and EPG of the offspring. The high dominance of AA, particularly in the membranes of the early placenta, may suggest that AA has an important role for placental development - organogenesis and vascularisation. Conversely. the enrichment of placental membranes with DGLA at term is more consistent with a profile favouring optimal blood flow to nourish the fetal growth spurt. On the other hand, maternal diabetes and preterm delivery led to a compromised M and DHA placental content. The experimental data strongly support the hypotheSiS that disturbance in utero environment through inadequate maternal essential fatty acid lipid nutrition impaired tissue synthesis DHA and/or AA in the offspring and. hence. may set the offspring at high risk for the development of cardiovascular disorders later in life.

Food labels are considered fundamental in communicating nutrition information and can assist consumers to make informed dietary choices. Recent legislative developments within the European Union (EU) have put food labelling high on the policy agenda with commentators calling into question how best to deliver nutrition communication to guide healthy food choices in the retail setting. In addition, the provision of nutrition information in catering outlets is currently being explored in the United Kingdom (UK). The current study examines the British consumers' use, perception and comprehension of nutrition information provision in retail and catering settings. The study methodology comprised of a short postal questionnaire (distributed across the UK) and eight focus group discussions (FGOs) (conducted in Scotland). The questionnaire was designed following a literature review of the topic area. Content validity, face validity, item analysis, readability, repeat and internal reliability were assessed and indicated the questionnaire was a valid and reliable instrument. The final tool contained 19 questions (49 items) and assessed frequency of reading food labels, perceived importance of food labels, regularity of dining out, desire to have nutrition information at catering outlets and ability to perform nutrition information tasks. The questionnaire was distributed to a sample representative of gender, age, country of residence and social background for the UK population. Responses from 786 adults indicated that the majority (>90%) of respondents read food labels, with 53.4% reportedly reading nutrition information on food labels 'frequently'. Ability to perform nutrition label tasks was good with 61.3% obtaining a high label performance score. Eating outside the home was common (42.0% reported eating out once a week or more and 75.3% reported eating out once a month or more). Respondents claimed to welcome nutrition information in the catering arena.FGDswere informed by the questionnaire results and conducted to elicit a detailed perspective on label comprehension and usage. A non-probabilistic sampling strategy was employed with participants purposively recruited to include those from the most and least deprived locations. Pre-existing groups (e.g. church, parent and community groups) were recruited across Scotland, with a combination of analytic frameworks utilised for data analysis. Findings from the FGDsindicated that motivation (to read labels and eat a healthy diet) was a key determinant of nutrition label reading behaviour. Knowledge of nutrition, time pressures and label reading practicalities were reported barriers to reading and using food labels. The impact of product price on label reading behaviour varied between affluent and non-affluent groups. Participants who used labels described doing so to make 'healthier' food choices and compare similar products. Lack of motivation to eat a healthy diet and interest yvere common reasons cited by participants for non-label use. The introduction of nutrition information in the catering setting received a mixed response. Many participants expressed concerns about the impact of nutrition information on their dining experience, others however accepted nutrition information could be of benefit to those consumers dining out frequently. In conclusion, a range of approaches can be identified to enhance nutrition communications (e.g. the use of innovative technologies and communication platforms, the provision of tailored nutrition education interventions, and the modification and simplification of information currently provided) although impact will be influenced by the wider marketing environment. For enhanced nutrition communications to be successful, consumers first need to be motivated to eat a healthy diet and to read and use nutrition information available on food labels.

No description available.

In a historical and quantitative study British food consumption patterns have been observed. The objective was to give a detailed description of the influence of ecological factors on attitudes to food choice. Food choice has been defined as the interplay of factors affecting the availability and acceptibility of food. Trends which have been identified in the availabilty of food are; the intensification of agriculture, the consolidation of the food industry, and the growth of niche markets. Trends which have been identified in the acceptibility of food are; a growth in manufactured food products, an increased awareness of the health value of food and a rising interest in food production methods with respect to the welfare of livestock and conservation of the environment. Three factors have been identified as being intrinsic to an ecological food ideology; method of food production, level of meat consumption and method of food preparation. The results show that the health value of foods has, to date, been the most significant influence on reducing meat consumption and the consumption of additive-free and organic foods, but strict vegetarianism is strongly connected with attitudes to animal welfare, and the consumption of organic food results also from a perception that organic food has superior taste. The results show also that consumers in the higher socioeconomic groups are most likely to express these influences in purchasing behaviour. However, sufficient interest was shown in welfare and organic foods by the Consumer Attitudes Survey to conclude that these foods now represent 'luxury foods'. n1 Conversely, for the group of organic food consumers in the Organic Survey, conservation of the environment was substantially the most important influence on food choice. In addition, respondentsin the Organic Survey exhibited very high levels of education suggesting that education is a significant factor as well as high income levels in determining purchasing behaviour, and in fact nearly half of the respondents in the Organic Survey were earning less than £250 gross per week. The results suggest that those consumers who are most strongly affected by concern for the environment, have not only made greater changes to their diet in terms of food intake than the national average, but also that these changes are closely aligned with current nutrition goals. In particular, low levels of meat consumption are accompanied by a high intake of fruit, vegetables and wholegrain cereal products. As the Taste Panel study showed, once the expectancy to eat meat at every meal has diminished, so the taste of non-meat products becomes more acceptable. Alternative approaches to agriculture and criteria for the determination of food quality are discussed. Food consumption patterns amongst the organic food consumers were more closely aligned to the potential agricultural output from an extensive ecological system of food production than from the current intensive approach to agriculture based largely on the use of grain for animal feed. The study believes therefore that Government policy should continue to support the Organic Sector, both practicising organic farmers and farmers wishing to convert. The study found little evidence however, to suggest that the very low level of consumption of industry prepared foods observed in the Organic Survey is likely to be adopted by the majority of the population.

In recent years, the relationship of gut hormones and adipocytokines with bone processes has received increased attention. It represents one aspect of the current skeletal investigation. The understanding of these interactions between bone metabolism and gastrointestinal or adipose tissue function must take into account homeostasis, appetite, neural system, and lifestyle. The proportion of bone mass to soft tissue is a relationship that seems to be controlled by delicate and subtle regulations that imply "cross-talks" between the nutrient intake and tissues like fat. Thus, recognition of the mechanisms that integrate a gastrointestinal-fat-bone axis and its application to a number of aspects of human health is vital for improving treatments related to bone diseases. This thesis analysed some of the aspects of experimental research regarding the effects of gut hormones and adiponectin when they were present in cell cultures of osteoblastic cell lines. mRNA expression levels of five gut hormone receptors were analysed in three osteoblastic cell lines (Saos-2, TE-85 and MG-63) using molecular biology techniques (reverse transcription and real time polymerase chain reaction, RT-qPCR). The gut hormone receptor mRNA expression was positive for GIPr, GHSr and GPR39 in the three cell lines whereas GLP-I r and GLP-2r were only expressed in the least mature cell lines (MG-63 and TE-8S). The responses to the gut peptides were studied finding significant changes in viability, and biochemical bone markers like alkaline phosphatase (ALP), procollagen type 1 amino-terminal propeptides (P 1NP), and osteocalcin production. Other experiments included the study of induction of e-fos in presence or absence of extracellular nucleotides using a luciferase assay and RT-qPCR. Adiponectin receptors and effects were investigated in one of the osteoblastic cell lines, Saos-2, finding the presence of the receptors in these cells. Functional studies revealed that adiponectin caused significant changes in viability, ALP and PI NP production. Significant changes were also observed in osteoprotegerin expression. The adiponectin receptors were additionally investigated in peripheral blood mononuclear cell (PBMC) cultures since they are the precursor for osteoclasts. The experiments were carried out in the presence or absence of cytokines (M-CSF, RANKL) which stimulate PBMC to commit to differentiation to osteoclastic cells. The levels of AdipoRI were higher than AdipoR2 when evaluated by qPCR. Immunofluorescence using anti-AdipoR 1 and -AdipoR2, displayed specific green emission in foci located mainly in the periphery of the cells but also in the cytosol in cells that were cultured up to 21 days with M-CSF and RANKL. These observations in immunofluorescence suggest that the adiponectin receptor could be undergoing redistribution in the osteoclastic cells. Taking together all the results it is enticing to speculate that the distribution of bone and soft masses is regulated by the direct effects of the hormones involved in the absorption of nutrients (gut hormones) and those produced in soft tissues like fat, as in the case of adiponectin.

The 21 s` century is witness to a dramatic rise in overweight, obesity and diabetes with its attendant social and economic costs. The influence of diet on a number of chronic diseases of ageing, including cardiovascular disease and osteoporosis, is now increasingly recognised. As a response to consumer demand for `health and wellness' products or `nutraceuticals', the food industry is investing heavily in functional `super' foods and supplements. To protect the consumer, foster innovation and harmonise the internal market, the EU has funded a number of research programmes to underpin its suite of legislative instruments, in particular the 2006 Nutrition and Health Claims Regulation. These include harmonisation of population reference intakes, dietary guidelines and maximum permitted levels of micronutrients in supplements and added to food. Entry to the EU register of approved health claims (structure/function and disease-risk reduction) on foods is through scientific substantiation procedures which follow a classical reductionist approach to biological science and epidemiology. However advances in genomic science are revealing the extent of inter-ancestral and inter-individual variation in response to, and requirements from, diet. The complex nature of the relationship between genome, diet and other environmental factors on health, highlights the inadequacies of traditional reductionist methodologies: new ways of looking at science and evidence through a stochastic prism are explored. As post-genome technologies refine and more precisely define health and dysfunction, the lack of any legal definition of `disease' in EU law threatens to blur an already opaque border between `food' and `medicine'. Where frameworks for population and personal nutrition and health guidelines are proposed, the reconciliation of food and medicine looks to lie in up-to-date biological definitions of `disease', `nutrient' and `essentiality'. Regulation of health claims and promotion of dietary guidelines will need to encompass the complexity of inter-ethnic and interindividual variation: this may necessitate use of appropriate qualifying or caveat statements (relating to action dynamics and/or weight of supporting evidence) fit for purpose in the new post-genome scientific reality. It will also demand new competencies of health professionals and regulators if the full potential of nutrigenomic advances for personal and public health is to be realised.

Recent studies have suggested that hepatic iron stores and the response to anaemia and inflammation are dependent on the mouse strains used and their Hfe status. Hepcidin and iron transporters such as DMT1 and Iregl are regulator molecules important in the pathology and physiology of Iron metabolism. The aim of these studies was to produce Hfe KO mice on a strain known to have higher basal hepatic iron levels and study iron homeostasis in relation to variable dietary iron and inflammation in these mice. The regulation of hepcidin was investigated in Huh7 cells exposed to various stimuli such as iron, inflammation and hypoxia. In addition, Huh7 cells were exposed to conditioned medium obtained from HH1 patient's macrophages. Results obtained from these studies are described and discussed in this thesis. In brief, it was found that the disruption of the Hfe gene causes a preferential accumulation of iron in the caudate lobe of the mouse liver. Contrary to previous findings, iron loaded diet does not have an effect on hepcidin expression in wt SWR mice while in Hfe KO SWR mice it significantly upregulates hepcidin expression. Furthermore, the upregulation of hepcidin by turpentine oil-induced inflammation in SWR mice is Hfe-dependent. Hepcidin expression is down regulated by hypoxia and anaemia and conditioned medium from patients with HH1 macrophages have no effect on hepcidin expression levels. In conclusion, the results show that SWR Hfe KO mice is not an accurate model for the study of HH1, the studies described here, however, contribute to the understanding of how hepatocytes respond to iron status, hypoxia and inflammation and the relevance of the Hfe gene in the regulation of hepcidin expression levels as well as hepatic DMT1 and Iregl expression levels.