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The past thirty years have seen a dramatic rise in obesity worldwide owing to a change in dietary composition, quantity of food consumed- positive energy balance, and a more sedentary life style. Accompanied with obesity is a chronic low grade inflammatory state defined by increased circulating cytokines and an increase in gene expression promoting inflammation. Multiple health risks are associated with obesity such as cardiovascular disease, insulin resistance, and type II diabetes. Advances in mass spectrometry have made wide scale proteomic studies possible and are redefining cell and molecular biology. One such area of that has become of considerable interest is protein acetylation which is observed in most cellular processes such as cell cycle regulation, gene expression, subcellular localization, metabolism, muscle contraction, protein stability, apoptosis, and more. Metabolic proteins are highly susceptible to acetylation with almost all showing the capacity to be acetylated. Our research, using an obese mouse model fed a chronic high fat diet and a lean control mouse model fed a standard chow diet, showed numerous differences in the acetylome between obese and lean animals in a fasted state. As well as, differences in the acetylome's of both animal models upon receiving a high fat meal. We showed that almost every mitochondrially located metabolic protein in obese animals is hyper-acetylated in a fasted state compared to lean animals and that upon feeding lean animals have a greater response in the change to their metabolic acetylome. We show that in the fed state lean and obese mice have almost completely different acetylomic profiles of mitochondrial and glycolytic metabolic proteins. Furthermore, we have observed possible new regulatory mechanisms utilizing acetylation to 1) determine the fate of the co-factor NADH in glycolysis and 2) control an ATP producing reaction in glycolysis. / Ph. D.

Background: Modifying the gut microbiota through the administration of probiotics during high fat feeding has been shown to attenuate weight gain and body fat accretion while improving insulin sensitivity in animal models. Objective: Our objective was to determine the effects of the probiotic VSL#3 on body weightand composition, skeletal muscle substrate oxidation, and insulin sensitivity and during 4 weeks of high-fat, hypercaloric feeding. We hypothesized that the probiotic would attenuate the body weight and fat gain and adverse changes in insulin sensitivity and substrate oxidation following high fat, hypercaloric feeding in young, non-obese males. Methods: Twenty non-obese males (18-30 y) volunteered to participate in the present study. Following a 2-week eucaloric control diet, subjects underwent a dual x-ray absorptiometry (DXA) to determine body composition, an intravenous glucose tolerance test (IVGTT) to determine insulin sensitivity, a skeletal muscle biopsy for measurement of substrate oxidation. Serum endotoxin was also measured. Subsequently, subjects were randomized to receive either VSL#3 (2 satchets) or placebo during 4 weeks of consuming a high fat (55% fat), hypercaloric diet (+1,000 kcal/day). Macronutrient composition of the high fat diet was 55% fat, 30% carbohydrate, and 15% protein. Results: There were no differences between the groups in subject characteristics or in the dependent variables at baseline. Body weight and fat mass increased less (P<0.045) following the high fat diet with VSL#3 compared to placebo. Insulin sensitivity (and other IVGTT variables) and both glucose and fat oxidation did not change significantly with time or VSL#3 treatment. Serum endotoxin concentration was not different between groups following the high-fat diet. Conclusions: VSL#3, a multi-strain probiotic, attenuated body weight and fat gain following a 4-week high fat, hypercaloric diet compared with a placebo. There were no differences between the VSL and control in circulating endotoxin, insulin sensitivity (and other IVGTT variables) or in skeletal muscle substrate oxidation. / Ph. D.

High-fat diets and obesity have been linked to unfavorable changes in gut bacteria and increased leakage of bacterially-derived lipopolysaccharide (endotoxin) from the intestinal tract into circulation, which is associated with low-grade inflammation, metabolic dysregulation and degradation of tight-junction proteins between intestinal cells. Probiotic supplementation is the practice of ingesting live strains of bacteria that are proposed to have a beneficial effect on the host by enriching the intestine with healthy bacteria. The purpose of this project was to determine if probiotic supplementation would prevent increased inflammatory tone, decreased oxidative capacity, and decreased tight-junction protein expression associated with high-fat feeding and elevated endogenous endotoxin. Male C57BL/6J mice were fed either a control (CD, 10% fat) or high-fat (HFD, 60% fat) diet for 4 weeks while receiving a daily oral gavage of water (C-VSL#3, HF-VSL#3) or probiotics (C+VSL#3, HF+VSL#3) equivalent to 1.2 billion live cultures. Changes in body weight, body composition, respiratory exchange ratio, energy expenditure, and glucose and insulin tolerance were measured in live mice. Markers of metabolic function were measured in whole muscle homgenates and mitochondria isolated from red and white skeletal muscle. Plasma endotoxin was measured in blood collected from fasted mice at the time of euthanization. The large and small intestines were collected and mRNA levels of tight-junction proteins and markers of nutrient sensing were measured. To determine a possible protective effect against endogenous LPS, a second cohort of mice were given an intraperitoneal injection of 0.1µg/kg LPS or saline to induce endotoxemia after four weeks of the aforementioned feeding protocol. Markers of metabolic function and inflammation were measured in mitochondria, skeletal muscle and liver. VSL#3 supplementation improved glucose homeostasis and markers of inflammation while enhancing nutrient sensing in the gut. / Ph. D.

Insulin resistance (IR), T2DM, and obesity together form a cluster of interrelated metabolic challenges that may be linked by metabolic inflexibility. Metabolic inflexibility is characterized by the resistance to switching substrate oxidation preference based on substrate availability and can be measured in either fasted or insulin-stimulated conditions. As the largest site for glucose disposal and a primary tissue influencing regulation of blood glucose concentrations, skeletal muscle likely plays a central role in regulating substrate oxidation preference based on substrate availability. Skeletal muscle lipotoxicity caused by an impaired regulation of fat uptake and oxidation is postulated to disrupt insulin signaling and lead to skeletal muscle IR. High dietary saturated fat intake results in reduced basal fat oxidation and a resistance to switching to carbohydrate oxidation during insulin-stimulated conditions in susceptible individuals. This metabolic inflexibility may lead to an accumulation of intramyocellular species that impair insulin signaling. Endurance exercise training improves the capacity for fat oxidation in metabolically inflexible individuals. However, relatively little is known about how endurance exercise training influences substrate oxidation preference when paired with a high fat diet (HFD). Therefore, the purpose of this study was to determine the effects of a HFD on substrate metabolism in skeletal muscle of sedentary and endurance trained (ET) males. Healthy, sedentary (n=17) and ET (n=7) males first consumed a 10-day moderate carbohydrate diet (55% carbohydrate, 30% total fat, <10% saturated fat) isocaloric to their individual energy requirements and then underwent a 4- hour high fat challenge testing session. During the session, they consumed a high fat meal (820 kcals; 25% carbohydrate, 63% total fat [26% saturated fat]), and skeletal muscle biopsies were taken in the fasted and 4-hour postprandial conditions. Participants then consumed a 5-day HFD (30% carbohydrate, 55% total fat, 25% saturated fat) and repeated the high fat challenge testing session. Substrate oxidation measures were performed on the collected skeletal muscle tissue, and the meal effect, defined as the percent change from the fasting to 4- hour postprandial condition, for each measure was calculated. There was a HFD by physical activity group interaction on meal effect for metabolic flexibility (P<0.05) and a HFD effect on meal effect for glucose oxidation (P<0.05). Meal effects for metabolic flexibility and glucose oxidation were maintained in the ET (20 ± 4% to 41 ± 21% and 128 ± 92% and 41 ± 15%, respectively; both P>0.05) but decreased in the sedentary (34 ± 7% to 4 ± 5% and 78 ± 26% to -21 ± 6%, respectively; both P<0.01) group. There were trends toward HFD effects on reductions in meal effects for total (P=0.062) and incomplete (P=0.075) fat oxidation, which were driven primarily by an increase in fasting total (12.1 ± 2.6 nmol/mg protein/h to 18.5 ± 2.3 nmol/mg protein/h; P<0.01) and incomplete (11.5 ± 2.5 nmol/mg protein/h to 17.6 ± 2.3 nmol/mg protein/h; P<0.01) fat oxidation in the ET group as a result of the HFD. Fasting total and incomplete fat oxidation did not change in the sedentary group (7.3 ± 0.8 nmol/mg protein/h to 7.8 ± 0.8 nmol/mg protein/h and 6.8 ± 0.7 nmol/mg protein/h to 7.2 ± 0.8 nmol/mg protein/h, respectively; both P>0.05). Overall, these findings suggest the ET state attenuates deleterious effects of a short-term HFD on reduced metabolic flexibility and insulin-stimulated glucose oxidation. In addition, a HFD-induced reduction in fat oxidation during the fasted-to-fed transition may be caused by differing mechanisms in sedentary and ET individuals. These findings provide a basis for future work targeting the elucidation of potential mechanistic differences in substrate oxidation preference between sedentary and ET individuals. / Ph. D. / Type 2 diabetes (T2DM) is a commonly occurring disease worldwide, and treatment of the disease is considerably burdensome for individuals and societies. T2DM is closely related to insulin resistance (IR) and obesity, and in each of these conditions, the characteristic of metabolic inflexibility has been observed. Metabolic inflexibility is a reduced ability to adjust fat or carbohydrate utilization for energy based on the availability of each of these macronutrients. Skeletal muscle may be an important tissue in the regulation of macronutrient utilization since it plays a key role in blood glucose regulation. High dietary saturated fat intake may lead to metabolic inflexibility in skeletal muscle in susceptible individuals. This metabolic inflexibility may result in increased storage of fat within skeletal muscle, which is hypothesized to disrupt insulin signaling. This disruption can lead to IR. Endurance exercise training improves metabolic flexibility. However, little is known about how endurance exercise training influences macronutrient utilization when paired with a high fat diet (HFD). Therefore, the purpose of this study was to determine the effects of a HFD on macronutrient utilization in skeletal muscle of sedentary and endurance trained (ET) males. Seventeen healthy, sedentary males and seven ET males first consumed a 10-day moderate-carbohydrate diet that was provided by the study investigators and designed to keep each participant weight stable. Participants then underwent a high fat challenge testing session in which they consumed a high fat meal and had skeletal muscle biopsies taken both before and after the meal. Participants then consumed a 5-day HFD, also designed to keep them weight stable, and repeated the high fat challenge testing session. Macronutrient utilization measures were performed on the collected skeletal muscle samples. Overall, metabolic flexibility was reduced in the sedentary group but was maintained in the ET group, which suggests that ET individuals may be protected against developing a HFD-induced metabolic inflexibility in skeletal muscle and its associated downstream negative effects on insulin signaling. In addition, fat utilization during the high fat challenge meal decreased in both sedentary and ET individuals as a result of the HFD. However, fat utilization in the fasted state was higher in ET individuals after the HFD compared with baseline, but fat utilization was the same in sedentary individuals before and after the HFD. This suggests there may be differences between sedentary and ET individuals in the mechanisms involved in the adjustment of fat utilization to dietary fat intake. Further research is needed to understand these differences, as they may play important roles in understanding how IR and T2DM develop.

Sammanfattning Syfte och frågeställningar Syftet med denna uppsats var att undersöka hurvida en LCHF (Low Carb High Fat) kost skulle kunna vara en hälsosam livsstil utifrån hur den påverkar olika hälsomarkörer hos normalviktiga personer där en viktminskning ej har eftersträvats. Frågeställningen var följande: Hur påverkar en LCHF-kost med maximalt 30 gram kolhydrater per dag under 3 veckor normalviktiga kvinnors BMR (Basal Metabolic Rate), blodlipider, blodglukos, blodtryck, kroppssammansättning, midje- och höftmått samt deras kroppsvikt? Metod En experimentell studie har genomförts på 8 normalviktiga kvinnor mellan 20 och 35 år. Samtliga försökspersoners BMR, blodlipider, blodglukos, blodtryck, kroppssammansättning, midje- och höftmått samt kroppsvikt mättes vid normala levnadsförhållanden innan studiens start samt efter 3 veckor av en strikt LCHF-kost med endast 30 gram kolhydrater per dag. Resultat BMR sänktes i genomsnitt med 4,4% vilket var signifikant (P&lt;0,05). HDL (High-density lipoproteins), LDL (Low-density lipoproteins) samt totalkolesterol höjdes signifikant (P&lt;0,01) med 21%, 25% respektive 22%. Ingen signifikant skillnad i LDL/HDL-kvot, triglycerider eller blodtryck erhölls. Blodglukos sänktes med 12%, skillnaden var dock ej signifikant. Andel kroppsfett sänktes signifikant med 8,7% (P&lt;0,01) och fettmassan med 11% (P&lt;0,01). Den fettfria massan visade ingen signifikant skillnad. Midjemåttet minskade signifikant med 2,3% (P&lt;0,05) och kroppsvikten i genomsnittt med 1,24 kg (P&lt;0,05). Höftmåttet visade ingen signifikant skillnad mellan mätningen före och efter 3 veckors LCHF-kost. Slutsats Studiens resultat tyder på övervägande negativa effekter av 3 veckors LCHF-kost på olika hälsomarkörer, vilket skulle kunna få allvarliga följder så som kardiovaskulära sjukdomar och depression förutsatt att verkan successivt ökar. / Abstract Aim The aim of this study is to examine if LCHF (Low Carb High Fat) could be a healthy lifestyle based on how it affects various health markers in normal weight people where weight loss has not been pursued. Specific question: How does intake of a LCHF diet with a maximum of 30 grams of carbohydrates per day during 3 weeks affect normal-weight women´s BMR, blood lipids, blood glucose, blood pressure, body composition, waist and hip dimensions and body weight? Method An experimental study was performed on 8 normal-weight women aged 20 to 35 years old. All subject´s BMR, blood lipids, blood glucose, blood pressure, body composition, waist and hip dimensions and body weight were measured at normal living conditions before the study began. Each subject was put on a strict 3 week Low Carb High Fat diet consisting of only 30 grams of carbohydrates per day. Results BMR was reduced by an average of 4,4% which was significant (P&lt;0,05). HDL, LDL and total cholesterol were increased significantly (P&lt;0,01) by 21, 25 and 22%, respectively. There were no significant differences in LDL/HDL ratio, triglycerides or blood pressure. Blood glucose concentration was reduced by an average of 12%, however the reduction was not statistically significant. Percentage body fat was significantly lowered with 8,7% (P&lt;0,01) and fat mass with 11% (P&lt;0,01). The fat free mass showed no significant difference between pre- and post-tests. The waist circumference was significantly decreased by 2,3% (P&lt;0,05) and average body weight of 1,24 kg (P&lt;0,05). The hip dimension showed no significant difference between pre- and post-tests.   Conclusion The results of this study indicate negative effects on the blood lipid profile after 3 weeks of LCHF diet, which could lead to serious consequences such as cardiovascular disease and depression provided that the effect is progressively enhanced.

The objectives were to investigate the effects of chromium (III) picolinate (CrPic; up to 2,000 ppb of Cr) on growing pigs subjected to a variety of stressors including thermal, immune, or dietary stress and to examine the effects of CrPic on steroidogenesis from adrenocortical cells. In the thermal stress study, high ambient temperature caused reduced weight gain and feed consumption (P andlt; 0.01), and low ambient temperature caused increased feed intake and feed:gain (P andlt; 0.01). However, these effects were not moderated by CrPic, and respiratory rate, plasma cortisol, or plasma glucose were unaffected by CrPic. In the immune stress study, pigs challenged with lipopolysaccharide (LPS) lost 951 g during 12 hours post injection, while the phosphate buffer saline (PBS) injected group gained 170 g (P andlt; 0.001). The LPS group showed higher rectal temperature (P andlt; 0.05), higher respiratory rate (P andlt; 0.05), greater plasma cortisol (P andlt; 0.001), and lower plasma glucose (P andlt; 0.05) than the PBS group. These effects were not ameliorated by CrPic. In the dietary stress study, pigs fed the high-fat diet (HFD) gained weight faster (P andlt; 0.05), consumed less feed (P andlt; 0.001), and had lower feed:gain (P andlt; 0.001). Plasma insulin concentration on d 14 decreased with CrPic (P andlt; 0.05) in a linear manner (P = 0.05). Consumption of the HFD resulted in increases of slaughter weight, perirenal fat, and back fat measurements (P andlt; 0.01). The CrPic resulted in linear reductions of carcass weight, last rib fat, last lumbar fat and average backfat (P andlt; 0.10). The effects of CrPic on carcass fat measurements were more significant in barrows than gilts. In the adrenocortical cell study, forskolin stimulated cortisol and DHEAs secretion from H295R cells. CrPic inhibited aspects of steroidogenesis in agonist-stimulated adrenocortical cells. Overall, dietary CrPic was unable to moderate the stress related effects due to high ambient temperature, low ambient temperature, or an endotoxin challenge. However, CrPic attenuated effects of HFD, mainly on body fat accretion of pigs, especially in barrows, and CrPic inhibited steroidogenesis in stimulated adrenocorticoid cells.

2 pp. / Of all the nutrients in the food supply, fat and cholesterol probably receive the most attention from health professionals and the public alike. The scientific evidence is clear that a high-fat diet relates to chronic health problems such as heart disease, some types of cancer, diabetes, and obesity. But both fat and cholesterol are natural components of the body that are vital to good health, and too little fat in your diet is just as unhealthy as too much. This article reviews dietary fats and provides guidelines for choosing foods to balance the type and amount of fat in your diet.

Sirtuins (SIRT) regulate the transcription of various genes involved in the development of diet-induced obesity and chronic disease; however, it is unknown how they change acutely following a high-fat meal. The purpose of this study was to determine the effect of a high-fat meal (65% kcals/d; 85% fat recommendation), on SIRT1-7 mRNA expression in blood leukocytes at 1, 3, and 5-h post-prandial. Men and women (N=24) reported to the lab following an overnight fast (>12H). Total RNA was isolated and reverse transcribed prior to using a Taqman qPCR technique with 18S rRNA as a normalizer to determine SIRT1-7 mRNA expression. An additional aliquot of serum was used to measure triglycerides. Data was analyzed using a RM ANOVA with P<0.05. Triglycerides (P<0.001; 124%) peaked at 3-h. SIRT 1 (P=0.004; 70%), and SIRT 6 (P=0.017; 53%) decreased expression at 3-h. SIRT4 (P=0.024) peaked at 5H relative to baseline (70%) and 3-h (68%). To our knowledge, this is the first study to report that consumption of a high-fat meal transiently alters SIRT mRNA expression consistent in a pattern that mirrors changes in serum triglycerides. Decrease in expression of SIRT1 and SIRT6 combined with an increased SIRT4 would be consistent with an increase in metabolic disease risk if maintained on a chronic basis.

Doctor of Philosophy / Department of Kinesiology / Craig A. Harms / The overall aim of this dissertation was to determine the impact of lifestyle (i.e. habitual and acute physical activity and diet), age, and sex on systemic and airway inflammation and oxidative stress. In study 1 (Chapter 2) we examined the impact of habitual physical activity level on the post-prandial airway inflammatory response following an acute bout of moderate intensity exercise. Results indicated that the mean exhaled nitric oxide (eNO; marker of airway inflammation) response increased for all groups at two hours post high-fat meal (HFM) (~6%) and returned to baseline by four hours post-HFM. However, there was a varying eNO response from baseline to four hours in the group that exercised in the post-prandial period compared to the group that remained sedentary. These findings suggest airway inflammation occurs after a HFM when exercise is performed in the post-prandial period, regardless of habitual physical activity level. In study 2 (Chapter 3) we investigated the post-prandial oxidative stress response to meals of varying calories and fat. Specifically, we assessed the post-prandial airway and systemic 8-isoprostane (a marker of oxidative stress) responses to meals with moderate-fat (8.5 kcal/kg of bodyweight) and high-fat content (17 kcal/kg of bodyweight) from baseline to six hours post-meal in a randomized crossover design. This study revealed that systemic 8-isoprostane increased from baseline to six hours post-meal (38.3%), but there was no difference between the moderate-fat meal (MFM) and HFM conditions. There were no changes in airway 8-isoprostane from baseline to six hours post-MFM or HFM, or between the MFM and HFM conditions. Lastly, in study 3 (Chapter 4), we were interested in examining 8-isoprostane responses in older adults, since 8-isoprostane has been reported to increase with age. Previous research also suggests that older women (OW) and older men (OM) have differences with regard to prevalence and severity of late-onset asthma. In this study, we sought to determine whether the airway 8-isoprostane response to a strenuous bout of exercise was different in OW compared to OM. A secondary aim was to determine whether post-exercise 8-isoprostane generation was correlated with decrements in lung function. Our results showed that the generation of 8-isoprostane from pre- to post-exercise increased ~74±77% in OW and decreased ~12±50% in OM. The decrease in 8-isoprostane generation was not correlated with improvements in lung function from pre- to post-exercise. These findings collectively contribute to the literature by enhancing our understanding of the impact of lifestyle factors, age and sex on modifying and potentially mitigating the risk of developing chronic diseases.