This study was carried out to investigate the fatty acid composition of tumoral, marginal and normal breast tissue in female breast cancer patients. Patients were recruited from St. Boniface General Hospital. A pre-operative blood sample was drawn. After surgery, sections were obtained from tumoral, marginal and normal breast tissues for histology and biochemical analysis. Extracted lipids from marginal tissue were significantly higher than those in normal or tumoral tissue. The lipid profile in tumoral tissue was significantly different in terms of fatty acid composition compared to normal and marginal tissue with less linoleic and alpha linolenic acid and more long chain polyunsaturated fatty acid of omega-3 and omega-6 series. Marginal tissue showed significantly less alpha linolenic acid compared to normal tissue. An inverse correlation existed between plasma level of 22:6 n-3 and breast cancer stage. We found different lipid profile in tumoral tissue compared to normal and marginal tissue.

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.

The key defects characteristic of hyperglycemia in T2D include increased hepatic glucose production, a diminution of insulin secretion, and an absolute impairment in peripheral insulin action. The objective of my thesis was to investigate the molecular mechanisms leading to fatty acid induced β-cell dysfunction and determine the role of a novel transcriptional coregulator in the regulation of hepatic glucose homeostasis. The first part of my work focused on the chronic effects of fatty acids on human pancreatic β-cell function. Using microarray technology I established an important role for fatty acids in the pathogenesis of β-cell dysfunction. Accordingly chronic exposure of islets to oleate resulted in a significant reduction in glucose-stimulated insulin secretion and to an increase in the rate of reactive oxygen species generation. Additionally, pre-treatment of human islets with oleate led to a significant increase in the rate of oxidation of this fatty acid and to a significant decrease in glucose oxidation. My data indicate that chronic exposure of human islets to fatty acids activates inflammatory and metabolic pathways that lead to oxidative stress. In addition, the first part of my work demonstrated that fatty acids induce oxidative stress in vitro an effect that is preventable to a large extent by the use of antioxidants. In this setting and recapitulating the human islet data, fatty acids are causally linked to impaired insulin secretion, and the induction of oxidative stress. Our report demonstrated that oxidative stress plays a key role in the decrease in β-cell function induced by chronic lipotoxicity. My work also demonstrated that fatty acids are causally linked to the induction of endoplasmic reticulum stress in human islets. Finally, in the second part of my work I provide novel evidence for the role of PHIP in the regulation of hepatic gluconeogenesis. My work is the first to demonstrate that PHIP suppresses hepatic gluconeogenesis in vitro and in vivo. PHIP is amongst the few proteins that have ever been reported to suppress gluconeogenesis to date. PHIP thus represents a novel target for pharmaceutical intervention of diabetes and the suppression of hepatic glucose production.

The effect of naturally produced volatile fatty acids (VFAs) on the removal of 2, 4-D from a wastewater during the denitrification process was studied in this thesis. The VFAs were generated from an anaerobic digester using soya flour solution as a synthetic feed. The digester was operated at an SRT and HRT of 10 days. The pH (4.8 ± 0.2) and temperature (32 ± 3 ℃) of the digester were not controlled. A mean VFA concentration of 3153 ± 801 mg/L was achieved with acid speciation results of acetic (51.4 %), propionic (27.5 %), n-butyric (19.6 %) and iso-valeric (1.4 %). The specific VFA production rate was 0.014 mg VFA/mg VSS/day. The extent of the digestion process converting the substrate from a particulate to soluble form was evaluated as the specific TOC solubilization rate (0.007 mg TOC/mg VSS/day), soluble COD production rate (0.022 mg SCOD/mg VSS/day) and percent VSS reduction (14 %). The low solubilization rate is possibly due to high feed solids (3.4%) which led to a heavily overloaded bioreactor. It also suggests that the particulate substrate was not entirely amenable to solubilization. The acclimation of 2, 4-D degrading bacteria was developed successfully in an SBR fed with sewage and 2, 4-D (30-100 mg/L) as carbon and energy sources. A mean MLSS of 3653 ± 547 mg/L and an SRT of 20 ± 9 days were observed during the research period. The settleability of the SBR sludge was excellent evidenced by a low sludge volume index (SVI) of 101 ± 50 mL/g and less than 5 mg/L of effluent suspended solids. The specific 2, 4-D degradation rate was 0.046 ± 0.018 mg/mg MLSS/day. However, the removal of 2, 4-D during 60 minutes of non-aerated phase was negligible while more than 90 % of the 2, 4-D was removed within 240 minutes of the aerated phase. The successful degradation of 2, 4-D is related to the length of the acclimation period, as the acclimation period increased, the specific biodegradation rate increased. A biosorption study using ultrasound pre-treatment of the SBR acclimated biomass suggested that less than 10 % of the removal of 2, 4-D was due to biosorption, while more than 90 % removal of the 2, 4-D was likely due to biodegradation. Denitrification batch tests (using SBR-acclimated biomass) demonstrated that the addition of a digester effluent rich in naturally-produced VFAs increased both the 2 specific denitrification rate and the 2, 4-D degradation efficiency, as compared to that using 2, 4-D as a sole carbon source. In particular, the specific denitrification rates increased from 0.0119 ± 0.0039 to 0.0192 ± 0.0079 to 0.024 ± 0.003 g NO₃-N/g VSS per day, when using 2, 4-D alone, 2, 4-D plus natural VFAs and natural VFAs alone as carbon sources. The percent 2, 4-D removal increased from 28.33 ± 11.88 using 2, 4-D alone as a carbon source to 54.17 ± 21.89 using 2, 4-D plus natural VFAs as carbon sources. The specific 2, 4-D degradation rate and 2, 4-D removal efficiency of unacclimated biomass were 2.0 to 2.5 times less than those of the acclimated biomass. Natural VFAs and synthetic VFAs were found to be identical in denitrification batch tests in terms of their use as a carbon source. The mean specific denitrification and VFA-C consumption rates as well as the mean specific 2, 4-D degradation rate derived from experiments using natural VFAs and 2, 4-D as carbon sources were close to the valuess from experiments using synthetic VFAs and 2, 4-D as carbon sources. Further exploration of 2, 4-D degradation behaviour with pulsed additions of NO₃-N did not find further significant 2, 4-D removal, although almost all of NO₃-N was used by the end of the experimental run due to endogenous carbon sources used for cell maintenance and growth. However, the higher the concentration of biomass used in the denitrification batch system, the larger the amount of 2, 4-D degraded and the faster the VFA-C and NO₃-N were consumed. Further research with respect to optimisation of the acid-phase anaerobic digestion process (e.g. to adjust SRT and HRT or to lower the solid content of synthetic feed) would improve the specific VFA production rate and the solubilization rate. More research on the SBR could be carried out to investigate its maximum 2, 4-D removal capability as well as the removal of other structurally related herbicides. Attempts could be made to stimulate the growth of denitrifiers in the SBR (e.g. to add certain amounts of NO₃-N according to proper C: N ratios or to increase the length of non-aerated time). More microbiological studies of 2, 4-D degrading bacteria may also be helpful to understand the combined SBR/denitrification and 2, 4-D degradation process. More theoretical aspects of modelling kinetics could be developed to apply the combined process in-situ at 2, 4-D contaminated sites.

No description available.

This study was designed to determine the distribution of fatty acids and cholesterol in total tumor and mammary tissues. Fatty acid profiles of phospholipid fractions from tumors and mammary glands were also determined. Fatty acids and cholesterols were analyzed by gas liquid chromatography. Methodology was developed for phospholipid separation by high performance liquid chromatography.Tumors derived from mammary glands in Strain A/ST mice were found to contain two to three times the amount of cholesterol compared to normal mammary glands. Mammary glands from safflower fed mice contained significantly higher percentages of linoleic acid. Linoleic acid content in stearicacid (SA-1) fed mice was sharply reduced. linoleic acid in mammary glands of animals fed a high fat Stearic acid (SA-4) , corn oil and stock diet fed animals displayed similar fatty acid profiles. Fatty acid analysis of tumors excised from mice fed the experimental diets showed similar patterns in comparison to normal mammary glands. The similar distributions were in the 18 carbon fatty acids. Distributions of phospholipid fatty acids in tumors and mammary glands were similar. Mammary gland phospholipids displayed increased percentages of short chain (14 carbons and under) fatty acids. Results of these studies demonstrate an increased availability of diet rich in polyunsaturated fatty acids.

In an effort to determine the metabolic changes that occur in fatty acid oxidation during the rapid growth period, the activity of carnitine palmitoyltransferase was measured in four groups of animals at 4, 8, 12 and 16 weeks of age.Muscle samples were taken from the soleus after animals were anesthesi2ed, and the samples were assayed for CPT activity. The CPT activity was significantly higher at 4 weeks than at 8, 12 and 16 weeks (p0.0001), and the fl week activity was significantly higher that the 16 week animals. It was found that there were two distinct phases of decline in CPT activity. The first occured from 4-8 weeks where a 74% drop in activity was recorded. The second phase was a more gradual decline that occured after 8 weeks. 21% decline in activity occured between 8 and 12 weeks, followed by a 23% decrease after 12 weeks. The first drastic drop in activity can be attributed to the residual heightened enzyme concentration carried over from the suckling period in which the rat was exposed to a high fat diet found in the mother's milk. The second phase of the decline in activity is due to a dietary change which caused a shift in metabolism from fat to glucose as the primary carbon source for fuel. / School of Physical Education

Phospholipase C mediated hydrolysis of the phosphoinositides yields second messenger products which have been linked to normal and abnormal cell growth processes. It has been suggested that fatty acids may regulate phospholipase C activity in vivo. This study sought to investigate relationships between phospholipase C activity, tumor size and dietary linoleic and stearic acids.Phospholipase C activity, of livers and serially transplanted mammary adenocarcinomas from female strain A/ST mice on one of three diets, was measured. Tumors ranged from .20 to 2.98 g at the time of removal. Phospholipase C activity of tumors was negatively and significantly (p<.Ol) correlated to increasing tumor weight in those diet groups that contained linoleic acid and no added stearic acid. The highest PLC activity was seen in small tumors across all diet groups. Maximum PLC activity of tumors was over 2 fold higher than the maximum activity of livers across all diet groups. PLC activity of the liver was negatively and significantly (p<.02) correlated to increasing tumor weight for samples from the high linoleic acid diet (SF-15). This research concludes that PLC activity varies during tumorigenesis and may reflect tumor development. / Department of Biology

Cyclopropene fatty acids (CPFA) found in cottonseed oil (CT) have been shown to reduce production of progesterone, a precursor of estrogen. Estrogenic hormones have been implicated in enhancing growth of mammary tumors. In this study, the effect of dietary cottonseed oil on estrogen production by mature female mice was determined by measuring urinary estrogen using High Performance Liquid Chromatography.At four months of age, five groups of three Strain A/ST female mice were placed on 20% fat diets containing 0, 2.5, 5.0, 10, and 20% cottonseed oil. The remainder of the fat in the experimental diets was corn oil sufficient to provide the balance of the 20% fat content in conjunction with other nutrients of equal percentages in all diets. At five day intervals mice were housed in metabolic cages and twenty-four hour urine samples were collected. Urine was purified on C18 columns and eluted with 1% phosphoric acid: acetonitrile: methanol: (54:35:11). Estrogen was quantified by High Performance Liquid Chromatography using a 250 X 5mm C18 column, hydrocortisone as an internal standard, and variable wavelength recorder set at 242 nm. The level of urinary estrogens after day 35 of the study was lower in all diets containing cottonseed oil. This is in agreement with several authors who have reported instances of physiological abnormalities in mammals which were fed increasing but low levels of dietary cyclopropenes. Since elevated estrogen levels have been identified as a risk for breast cancer, this study examines the relationship between dietary cyclopropenes and estrogen hormone production in strain A/St mice. / Department of Biology

To investigate the possible changes in pyruvate oxidation during the rapid growth period in an animal model, the oxidation of radioactive labeled pyruvate was measured in mitochondria isolated from the gastrocnemius muscle of Sprague--Dawley rats between 4 and 16 weeks of age. The influence of the fatty acid derivative palmitylcarnitine, as an inhibitor of pyruvate oxidation, was also tested.The gastrocnemius muscle was removed from anesthesized animals at 4, 8 and 16 weeks of age. Isolated mitochondria from the muscle samples were incubated with C1--14C] pyruvate and E1-14C] pyruvate + palmitylcarnitine in a KC1 medium. The decarboxylation of pyruvate was measured by the evolution of radioactive labeled carbon dioxide. Pyruvate oxidation significantly (p .; 0.0001) increased from ages 4 to 16 weeks. The initial low rate of pyruvate oxidation was attributed to the residual metabolic effects of the pre-weaned animal' high-fat diet. The subsequent increase in the capacity of pyruvate oxidation was then explained by the shift in the animaldiet to high-carbohydrate lab chow. These results may also be attributed to the maturation of the hindlimb muscle fibers during this period: the differentiation of predominately red, oxidative fibers to an increase in the percentage of white, glycolytic fibers, common in the adult hindlimb. The fatty acid derivative, palmitylcarnitine, failed to inhibitpyruvate oxidation at the level of decarboxylation. This finding supports the proposal that fatty acids do not inhibit glucose oxidation directly, but instead suppress glycogen breakdown. Thus, the findings indicate an increase in the capacity for- pyruvate oxidation during the rapid growth period without inhibition by the fatty acid derivative, palmitylc_arnitine. / Department of Biology