Dietary fatty acids are precursors of the lipid mediator group of compounds termed fatty acid ethanolamides (FAE). Prolonged intake of specific types of dietary fats has been shown to increase FAE levels. However, the short term effects of qualitative dietary fat intake on FAE levels remain understudied. Hence, the objective of this study was to identify the effect of diets containing varying concentrations of n-9 from canola oil (CO) and n-3 fatty acids from DHA rich oil (DRO) on plasma and organ FAE levels after different time points in male Sprague Dawley rats. Sixty-four rats were randomly assigned into four groups and were fed diets containing 40% as energy of either safflower, 95% CO: 5% DRO, 50% CO: 50% DRO and 5% CO: 95% DRO. These diets were consumed within a 2hr window in all groups. Circulating fatty acid and FAE levels were measured at 3, 6, 12 and 24hr within each group. At 3hr, significant differences (p<0.05) in plasma oleoylethanolamide (OEA) levels were seen in the 95% CO group: 5% DRO group and 5% CO group: 95% DRO group as well as between 50% canola oil group: 50% DRO and 5% CO group: 95% DRO. In all dietary groups, palmitoylethanolamide (PEA) levels were not significantly different at 3, 6 and 24hr compared to 0hr, but did at 12hr where the 50% CO:50% DRO group showed significantly lower levels than seen in the 95% CO group, but PEA levels were not different from the 5% canola oil group. Although plasma FAE levels were generally multiple times lower than observed in small intestine, liver or brain, arachidonoylethanolamide (AEA) levels were significantly lower in the 95% DRO group than in the remaining two groups. Plasma docosahexanoylethanolamide (DHEA) showed no difference across all time points except at 24hr where levels were higher (p<0.05) in the 95% DRO group than in the remaining two groups. In liver at 3hr, OEA levels were higher (p<0.05) in the 95% CO group than the groups with lesser concentrations of oleic acid, while liver OEA levels showed no difference at any other time points across dietary groups. LEA levels were higher in 95% CO: 5% DRO group compared to the 5% CO group: 95% DRO group after 3hr of feeding. Liver DHEA levels were observed to be highest in the 5% CO group: 95% DRO group at 3 and 12, but not at 6 or 24hr. The dietary fatty acid composition affects plasma and organ fatty acid profiles in a time dependent manner and also produces time shifts in plasma and organ FAE levels. These dietary induced changes according to time points in the levels of FAEs may translate into discernible changes in energy expenditure and lipid levels which may in turn influence the risk of obesity. / February 2017

Replacement of saturated fatty acids (SFA) with monounsaturated fatty acids (MUFA) or polyunsaturated fatty acids (PUFA) impacts risk of atherosclerosis and cardiovascular disease (CVD). However, although dietary fatty acids (DFA) have been established as an important factor related to CVD, their exact mechanisms of action have not been clearly established. One of the possible mechanisms is that DFA convert to fatty acid ethanolamides (FAEs), such as oleoylethanolamide (OEA), palmitoylethanolamide (PEA) and arachidonoylethanolamide (AEA), which are thought to associate with lipid signalling, fat oxidation and appetite control. Hence, the objectives of this thesis were to identify the impact of diets containing corn oil, canola oil, DHA + canola oil and fish oil on plasma and organ levels of FAEs as well as energy metabolism and lipid profiles in Syrian Golden hamsters. Forty-eight hamsters were provided diets containing 6% treatment oil for 30 d before sacrifice. Across all diets, in proximal small intestine and liver, animals fed canola oil showed higher (p<0.05) levels of OEA than corn oil and fish oil fed groups, but no difference compared to those fed DHA +canola oil. In plasma, fish oil fed animals showed higher (p<0.05) OEA and PEA levels and lower (p<0.05) AEA levels compared to all other groups. Feed intakes (g/d), oxygen consumption (ml/g) and body composition of total fat (%) and mass (g) did not differ across groups. However, energy expenditure associated with fat oxidation (%) was higher (p<0.01) in canola oil and DHA + canola oil fed hamsters compared to those consuming corn oil and fish oil. Also, body composition of fish oil fed animals showed a lower (p<0.01) total lean mass (g) compared to other three groups and a lower (p<0.01) total mass (g) compared to DHA + canola oil diets, but no difference compared to animals fed the canola oil diet. None of the treatments had any effect on triglyceride (TG) or C-reactive protein (CRP) levels. The fish oil group showed a higher (p<0.01) plasma total cholesterol (TC) levels than all other three groups. No differences existed between DHA + canola oil and fish oil groups in HDL or Non-HDL levels, but these levels were different (p<0.01) compared to corn oil group and canola oil groups. To conclude, different DFA affect whole body energetics and plasma lipid profiles. Also DFA produced marked shifts in plasma and organ levels of OEA, PEA and AEA. These dietary induced shifts in FAEs may translate into discernable changes in energy expenditure and lipid levels which in turn influence CVD risk.

Replacement of saturated fatty acids (SFA) with monounsaturated fatty acids (MUFA) or polyunsaturated fatty acids (PUFA) impacts risk of atherosclerosis and cardiovascular disease (CVD). However, although dietary fatty acids (DFA) have been established as an important factor related to CVD, their exact mechanisms of action have not been clearly established. One of the possible mechanisms is that DFA convert to fatty acid ethanolamides (FAEs), such as oleoylethanolamide (OEA), palmitoylethanolamide (PEA) and arachidonoylethanolamide (AEA), which are thought to associate with lipid signalling, fat oxidation and appetite control. Hence, the objectives of this thesis were to identify the impact of diets containing corn oil, canola oil, DHA + canola oil and fish oil on plasma and organ levels of FAEs as well as energy metabolism and lipid profiles in Syrian Golden hamsters. Forty-eight hamsters were provided diets containing 6% treatment oil for 30 d before sacrifice. Across all diets, in proximal small intestine and liver, animals fed canola oil showed higher (p<0.05) levels of OEA than corn oil and fish oil fed groups, but no difference compared to those fed DHA +canola oil. In plasma, fish oil fed animals showed higher (p<0.05) OEA and PEA levels and lower (p<0.05) AEA levels compared to all other groups. Feed intakes (g/d), oxygen consumption (ml/g) and body composition of total fat (%) and mass (g) did not differ across groups. However, energy expenditure associated with fat oxidation (%) was higher (p<0.01) in canola oil and DHA + canola oil fed hamsters compared to those consuming corn oil and fish oil. Also, body composition of fish oil fed animals showed a lower (p<0.01) total lean mass (g) compared to other three groups and a lower (p<0.01) total mass (g) compared to DHA + canola oil diets, but no difference compared to animals fed the canola oil diet. None of the treatments had any effect on triglyceride (TG) or C-reactive protein (CRP) levels. The fish oil group showed a higher (p<0.01) plasma total cholesterol (TC) levels than all other three groups. No differences existed between DHA + canola oil and fish oil groups in HDL or Non-HDL levels, but these levels were different (p<0.01) compared to corn oil group and canola oil groups. To conclude, different DFA affect whole body energetics and plasma lipid profiles. Also DFA produced marked shifts in plasma and organ levels of OEA, PEA and AEA. These dietary induced shifts in FAEs may translate into discernable changes in energy expenditure and lipid levels which in turn influence CVD risk.

It is well established that replacing dietary saturated fatty acids with unsaturated fatty acids reduces cardiovascular disease (CVD) risk. Although epidemiological and clinical evidence indicate health benefits of consuming various fatty acid classes including n-9, n-6, and short- and longer-chain n-3 fatty acids, current dietary recommendations fall short of providing the optimal amounts of these fatty acids in daily diets. In addition, significant knowledge gaps remain in our understanding of the effects of, and mechanisms underpinning the action of, the various fatty acid classes on risk factors for CVD. The objective of this research was to contribute to the evaluation of health benefits of using different dietary oils, and determine how these benefits may play a role in improving public health and decreasing CVD risk. Additionally, this research examined effects of diet-gene interactions, endogenous fatty acid ethanolamides (FAEs) on body fat mass distribution as well as changes in the composition of gut microbiota following consumption of dietary oil treatments. The Canola Oil Multicenter Intervention Trial (COMIT) was conducted as a 5-phase randomized, controlled, double-blind, cross-over clinical trial. Each 4-wk treatment period was separated by 4-wk washout intervals. A total of 130 volunteers with abdominal obesity consumed each of 5 identical weight-maintaining, fixed-composition diets with one of the following treatment oils (60 g/3000 kcal) in the form of beverages: 1) conventional canola oil (Canola; n-9 rich), 2) high–oleic acid canola oil with docosahexaenoic acid (CanolaDHA; n-9 and n-3 rich), 3) a blend of corn and safflower oil (25:75) (CornSaff; n-6 rich), 4) a blend of flax and safflower oils (60:40) (FlaxSaff; n-6 and short-chain n-3 rich), and 5) high–oleic acid canola oil (CanolaOleic; highest in n-9). At endpoints, plasma fatty acid levels reflected the differences in fatty acid composition of five dietary treatments. All diets lowered total cholesterol (TC) compared with baseline. TC was lowest after the FlaxSaff phase and highest after CanolaDHA. The CanolaDHA treatment improved HDL-C, triglycerides, and blood pressure thereby reducing Framingham risk scores compared with other oils varying in unsaturated fatty acid composition. Homozygotes minor allele carriers of rs174583 (TT) on FADS2 gene showed lower (P<0.01) plasma EPA and DPA levels across all diets, but no differences were observed in DHA concentrations after the CanolaDHA feeding. In addition, plasma FAE levels were positively associated with plasma fatty acid profiles. Minor allele A carriers of rs324420 of FAAH gene showed a higher (P<0.05) plasma FAE levels compared with major allele C carriers across all diets, and showed higher (P=0.0002) docosahexaenoylethanolamide levels in response to the CanolaDHA diet. Impacts of consuming 60 g of five dietary oil treatments on gut microbiota composition were relatively minor at the phylum level and mainly at the genus level, while BMI contributed to a significant shift at the phylum level. In conclusion, consumption of a novel DHA-enriched canola oil improved blood lipid profile and largely reduced CVD risk. Diet-gene interactions might help identify sub-populations who appear to benefit from increased consumption of DHA and oleic acid. The metabolic and physiological responses to dietary fatty acids may be influenced via circulating FAEs, while the altered microbiota profile by shifts in MUFA and/or PUFA may be associated with specific physiological effect. Personalized diets varying in unsaturated fatty acids composition based on specific lifestyles, environmental factors, psychosocial factors, and genetic make-ups will become the future “healthy eating” recommendations to prevent CVD risk. / May 2016