The aim of this thesis was to investigate how dietary fatty acids affect the risk for cardiometabolic disease, i.e. cardiovascular disease (CVD), type 2 diabetes and obesity. The overall hypothesis was that unsaturated fatty acids and especially the predominant polyunsaturated fatty acid (PUFA) linoleic acid (LA), 18:2n-6, would decrease cardiometabolic risk compared with saturated fatty acids (SFAs), in line with current recommendations to partly replace dietary SFA with PUFA. Papers I and V were observational studies based on the community-based cohort Uppsala Longitudinal Study of Adult Men (ULSAM). Adipose tissue fatty acid composition was determined as biomarker for dietary fat intake. Studies II, III and IV were randomised short-term interventions on human volunteers, in which different dietary fats were provided to the participants. In 71-year-old men, adipose tissue LA and α-linolenic acid (18:3n-3) were associated with insulin sensitivity (euglycaemic clamp), although this association was diminished for LA after adjusting for lifestyle variables. Different SFA displayed divergent associations; only palmitic acid (16:0) was inversely associated with insulin sensitivity (Paper I). In Cox regression analyses, LA was modestly associated with decreased all-cause mortality, but not CVD mortality during 15 years follow-up (Paper V). In a 3+3-week cross-over study on 20 weight-stable volunteers with dyslipidaemia, all foods were provided. A rapeseed oil-based diet distinctly lowered low-density lipoprotein cholesterol and triglycerides compared with a dairy-fat based diet (butter, cream and fatty cheese). Insulin sensitivity or coagulation factors were not affected (Paper II). In a 10-week randomised trial on 67 abdominally obese participants, PUFA (mostly sunflower oil) decreased liver fat compared with SFA (mostly butter) under isocaloric conditions. In individuals considered highly compliant to study diets, lipoproteins were also decreased during the PUFA diet (Paper III). In a 7-week double-blind randomised trial on 41 healthy volunteers, PUFA (sunflower oil) decreased the total:HDL cholesterol ratio compared with SFA (palm oil) during moderate weight gain (1.5 kg) (Paper IV). In conclusion, LA (PUFA) intake is associated with decreased cardiometabolic risk compared with higher SFA intake, overall supporting a beneficial role of non-tropical vegetable oils in place of solid fats in preventing fatty liver and cardiometabolic disorders.

In an effort to determine the relative amount of carbohydrates and free fatty acids (FFA) utilized by trained and untrained men at a submaximal workload and the relationship of selected mitochondrial and glycolytic enzyme activities, six trained and six untrained males were exercised for one hour at 50o of their maximal oxygen uptake (V02 max).Muscle samples were taken from the vastus lateralis in three cyclists and from the gastrocmemius in three runners from each group before and after exercise, and later assayed for succinic acid dehydrogenese, malic acid dehydrogenase, lactic acid dehydrogenase, phosphorylane, and carnitine paLmityltransf erase activities, and glycogen levels. Th e post exercise sample was assayed for glycogen, another was sectioned and stained for glycogen and fiber composition (PAS and-glycerolphosphate dehydrogenase) and from a third coupled mitochondria were isolated to determine 14'CO2 production from oxidation of varied levels of 14C-palmityl-CoA. Blood drawn before, immediately after, and thirty minutes after the exercise was assayed for FFA, glycerol, triglycerides, and glucose levels. Activities of oxidative enzymes (SDH, MDH, and CPT) were significantly higher (2-3 times greater) in the trained individuals than in the untrained group. Glycolytic enzyme activities were higher in the untrained group, probably due to higher fast twitch fiber populations. During exercise, FFA levels rose to the same degree in both groups; however, glycerol levels increased almost five times greater during the exercise in the trained subjects indicating the FFA turnover was much larger in the trained subjects. Blood glucose levels increased by an average of 11 mg% during the exercise in the trained subjects but fell by 8 mg% in the others, suggesting a greater selective uptake by the untrained subjects. Muscle glycogen depletion was 66% greater in the untrained group. These catabolic processes were independent of muscle fiber type, indicating that aerobic training increases those enzyme activities associated with FFA oxidation. Trained individuals are thus able to shift to fatty acids as the primary carbon source for the citric acid cycle, sparing glycogen during submaximal work.

Trophic ecology is a key component in describing patterns of variation between and within populations, particularly in Arctic marine systems wherein climate change is impacting food webs. This thesis investigates the trophic ecology of Arctic char (Salvelinus alpinus) in the Cumberland Sound region using a multi-indicator approach. My data show trophic niche differences between resident and anadromous ecotypes and evidence for estuarine feeding by residents. I document a shift in the marine diet of Arctic char from zooplankton to capelin (Mallotus villosus) – a novel prey species in this region – that has occurred in less than a decade. Changes in Arctic char growth imply population-level effects of this shift; however, more research is required. Finally, I find lipid effects on δ13C and lipid-extraction effects on δ15N and δ34S for Arctic char muscle tissue. Lipid-correction models did not provide adequate δ13C estimates; thus, chemical extraction or ecotype-specific validation of models is recommended.

No description available.

The discovery of new immunosuppressive drugs has resulted in an improvement of short-term graft survival. Despite this achievement, long-term cardiac allograft survival has not been correspondingly improved. Cyclosporine A (CsA), an effective immunosuppressive drug, has been shown to increase the risk of hyperlipidemia, hypertension, kidney injuries and chronic rejection despite its extensive use in the clinical setting. Therefore, these side-effects of CsA, may further contribute to graft failure over long-term. Early studies have shown that fish oil may reduce side-effects of CsA. These beneficial effects of fish oil may be related to n-3 fatty acids (n-3 FAs) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Flaxseed oil is another major source of an n-3 FA, namely α-linolenic acid (ALA). However, its impact on heart transplantation has not been fully explored. The current study aimed to investigate whether dietary flaxseed oil and fish oil reduce post-transplant complications and prolong graft function in a rat cardiac allograft model. Male Fischer and Lewis rats were used as donors and recipients, respectively, to generate a heterotopic cardiac allograft model. After transplant, animals were randomly assigned into 3 groups and fed a diet supplemented with: a) 5% w/w safflower oil (control n=7), b) 5% w/w flaxseed oil (n=8) or c) 2% w/w fish oil (n=7) and an intraperitoneal injection of cyclosporine A (CsA) (1.5 mg/kg/d) over 12 weeks. Body weight, blood pressure (BP), plasma levels of lipids, CsA, and select cytokines, fatty acid profile of hearts (native and graft) and liver tissues as well as graft function and chronic rejection features were assessed. Body weight and blood CsA levels were similar among the groups. As compared to controls, both diet treated groups demonstrated a significantly lower systolic blood pressure (SBP) (p<0.001), diastolic blood pressure (pressure (DBP) (p<0.001), mean arterial pressure (MAP) (p<0.001), heart rate (p<0.05), abdominal fat (p<0.05) and plasma levels of macrophage chemoattractant protein-1 (MCP-1) (p<0.05). Moreover, the fish oil group had significantly (p<0.05) lower plasma levels of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL), as compared to the control group. High-density lipoprotein cholesterol (HDL) concentrations were significantly higher (P<0.05) in the flaxseed oil-treated group as compared to the other two groups. Data of this study suggest that both flaxseed oil and fish oil may provide similar biochemical, hemodynamic and inflammatory improvements after heart transplantation; however, these apparent beneficial changes were not accompanied with significant reductions in chronic rejection states or apparent histological evidence of cyclosporine-induced nephrotoxicity in this model.

Heat exposure causes several physiological and metabolic alterations. Although lipids are vital in sustaining energy production, heat-induced alterations in lipid metabolism have not been clearly established. CHAPTER 1 reviews the known metabolic alterations resulting from heat stress, with a specific focus on changes in whole-body lipid utilization and plasma lipids. CHAPTER 1also outlines the physiological changes caused by heat stress, and their role in reducing exercise performance. The study presented in CHAPTER 2 has shown that, compared to thermoneutral conditions, NEFA concentrations were 37% higher following passive heating and 34% higher following exercise in the heat, without significant changes in whole-body lipid utilization. In addition, the level of hyperthermia attained during passive pre-heating and exercise in the heat resulted in a 13% decrease in total external work and a significantly higher rate of perceived exertion. CHAPTER 3 summarizes the study results and presents the limitations and applications of the study.

Current trends toward warmer air temperatures and longer ice free seasons in Hudson Bay are expected to cause changes in Arctic marine ecosystem dynamics. Ringed seals (Phoca hispida) will likely experience changes in levels of predation, competition, and prey availability. The purpose of this thesis was to investigate seasonal, inter-annual, and spatial variation in Hudson Bay ringed seal feeding ecology. Fatty acid composition, δ15N, and δ13C varied significantly by season, suggesting seasonal changes in foraging habitat and diet. Spatial differences in ringed seal stable isotope ratios occurred between western and eastern Hudson Bay, and there was a strong relationship between spring air temperature and δ15N. Peak δ15N occurred within a range in spring air temperatures between approximately -5°C and -2°C. I propose that the high δ15N observed in ringed seals within this temperature range is indicative of relatively greater importance of capelin (Mallotus villosus) in the ringed seal diet.

Narwhals (Monodon monoceros) are sentinel species in the Arctic environment and are a vital component for Inuit culture and subsistence. The Arctic is undergoing rapid changes in temperature and sea ice cover and relatively little is known about how this has and will change narwhal foraging behaviour. There are three narwhal populations in the world, the Baffin Bay (BB), Northern Hudson Bay (NHB), and East Greenland (EG) populations; however, foraging behaviour, in terms of dive behaviour and primary dietary components, has really only been investigated in the BB population. Using a combination of stable isotopes, fatty acids, genetic techniques, and satellite tracking technologies I evaluated foraging behaviour in all three of the world’s narwhal populations. I also investigated social structure in the BB population to determine how adaptable narwhals are to a changing and dynamic Arctic environment. Stable isotopes (δ13C and δ15N) and fatty acids are chemical signatures in the tissues of an organism that can provide long-term information on their diet over varying temporal scales depending upon the tissue. Stable isotope analysis in the three narwhal populations found they forage on different primary prey, suggesting narwhal are adaptable in their preferred prey and that there is potential for them to adjust foraging behavior in the face of changing climate. Dietary changes were also assessed over three decades to determine how sea ice changes have affected narwhal foraging for the NHB and BB populations. Dietary changes were evident and can be attributed to changes in sea ice patterns and an altered migratory pathway for narwhals. An understanding of narwhal social structure is also needed to determine how behaviourally flexible narwhal are in diet and site fidelity. Genetic relatedness and dietary signatures from fatty acids were assessed for an entrapped group to determine if individuals that are closely related forage together, which would support a matrilineally driven social structure where females teach their young foraging strategies, and/or travel and forage together. I found no evidence that narwhals form a matrilineal social group, but they may display a fission-fusion structure, which may be an adaptation to patchy prey distribution in the Arctic. Finally, narwhal dive behaviour in all three populations was investigated to determine if dive behaviour could be used to predict diet. Dive differences among populations did correspond with differences in diet, suggesting that narwhals employ specialized foraging strategies. This has repercussions for their ability to adapt to ecosystem changes. Overall, narwhals may be more flexible in terms of their foraging behaviour than previously believed. However, an increased resilience to changing food webs will not be the only predictor of how narwhals will fare in the face of a changing climate; how they respond to increased industrial activities in their preferred habitats, increased predation from southern predators, and increased competition from southern cetaceans and humans alike, will play an equally large role in how they cope with the future.

This thesis presents evidence for a novel mechanism of regulation of pyruvate dehydrogenase (PDH) kinase by fatty acids and also results of a study of muscle triacylglycerol concentration. In animals regulation of PDH complex activity is central to the selection of respiratory fuels and to the conservation of glucose during carbohydrate deprivation. The principal means of regulation of PDH complex is interconversion of phosphorylated (inactive) and dephosphorylated (active) forms effected by PDH kinase and PDH phosphatase. Earlier in vitro studies by others had identified both shorter term (min) and longer term (hours) mechanisms of activation of PDH kinase by fatty acid. In the present study PDH kinase activity (as measured by rates of ATP-dependent inactivation of PDH complex in extracts) was shown to be increased when rat heart mitochondria were incubated with palmitoyl-L-carnitine [PC] (and other CoA utilising respiratory substrates). The activation of PDH kinase persisted through removal of respiratory substrate following incubation with CCCP. A comparable effect of PC was also demonstrable in heart mitochondria from 48h-starved rats (i.e. the mechanism may be distinct from that which increases PDH kinase activity in starvation). Rates of ATP-dependent inactivation of PDH complex were also increased when extracts of rat heart mitochondria were incubated with palmitoyl-CoA (PCoA); the increase was comparable with that seen on incubation of intact mitochondria with PC. The PC effect in intact mitochondria and the PCoA effect in mitochondrial extracts may not be identical as PCoA further increased PDH kinase activity in extracts from mitochondria incubated with PC. Rates of incorporation of ³²P from [γ-<sup>32</supP]ATP into PDH complex were unaltered by pnor incubation of mitochondria with PC or by pnor incubation of mitochondrial extracts with PCoA. Three lines of evidence confirmed that the effect of PC to accelerate ATP-dependent inactivation involved phosphorylation of the PDH complex (viz; use of a non-phosphorylatmg ATP analogue; use of known inhibitors of PDH kinase; and use of known activators/inhibitors of PDH phosphatase). Earlier studies had shown that phosphorylation in punfied bovine and porcine PDH complexes is half site (involves only one α-chain in E1 (α2β2) and had suggested that phosphorylation in rat heart complex may be full site (i.e. involves both α-chains). The present study suggests the possibility that elevation of fatty acyl CoA under slaughter house conditions might be a determinant of half site phosphorylation. A method was developed and evaluated for measurement of triacylglycerol in rat soleus muscle strips with the object of investigating factors that may regulate triacylglycerol synthesis in this muscle. This study was abandoned because, although the method was highly reproducible, great variation was found in the triacylglycerol concentration of individual muscles suggesting the possibility of variable contamination with small amounts of adipose tissue.

Strawberry (Fragaria ananassa) is an economically important soft fruit that is highly valued as a fresh product and flavouring. During ripening, strawberry fruits undergo a number of physiological changes affecting colour, texture and flavour. An understanding of these changes at the biochemical and molecular level will be important in developing strategies for enhancing the quality attributes of this fruit. A cDNA encoding a ripening- enhanced acyl carrier protein (RE-ACP) was previously isolated from strawberry fruit. AC? is an essential component of fatty acid synthesis in both plants and animals. The aims of this thesis were to isolate and characterise this and other members of the ACP multigene family expressed in strawberry fruit. Six closely related putative AC? cDNA isoforms were identified from strawberry. Two of these were obtained by screening a cDNA library from ripe fruit and three were obtained by a technique known as candidate fragment length polymorphism (CFLP) that utilised ACP gene-specific primers for AFLP-cDNA display. Northern analysis was not able to differentiate their expression but ACP was highly up-regulated in ripening fruit whereas low levels of expression were detected in other strawberry tissues, including achenes (seeds), expanding leaves and flowers. The RE-ACP was over-expressed in E. coli and the recombinant protein partially purified. The over-expressed protein had a M(_r) of 20kDa on SDS-PAGE and appeared to form a dimer. A genomic library was constructed from F. ananassa from which two different genomic clones closely related to RE-ACP were obtained. Promoter analysis indicated the presence of regulatory elements. The characterization of putative ACP cDNA and genomic clones, including the 5' upstream regions, is described and their possible role in strawberry fruit is discussed. Key words: Strawberry, fruit, ripening, gene expression, genomic, cDNA, fatty acid, acyl carrier protein, aroma, promoter.