Search

Added sugar is sweetener added to foods during processing or preparation that offers no health benefits to the consumer. The mean U.S. intake of added sugar is ~16% of total calories; at the highest level of consumption, this value exceeds 35%. In addition, 78% of added sugar typically consumed is refined from C4 plants (e.g., corn and sugar cane) and it follows that the d13C value of these sweeteners is conspicuously high compared to carbohydrates derived from C3 plants. We first suggested in 2006 the potential for the d13C of human tissues to indicate corn- and cane-sugar intake for use in a clinical setting. At present, self-reported dietary assessment methods are commonly used to measure added sugar intake, but are subject to underreporting, particularly for sugar-rich foods. If a carbon isotope technique could produce a quantitative indicator of dietary sugar intake, it would be invaluable to the prevention and clinical treatment of chronic diseases associated with excess sugar consumption. Research to date has focused upon testing the correlation between diet as characterized either by bulk food d13C value or by food composition (e.g., added sugar intake quartile) and the d13C values of human tissues. Analysis of hair, nail and red blood cells in modern humans with known diet has revealed associations between the d13C value of bulk diet and the d13C value of these tissues. With respect to added sugar, the d13C values of blood serum and fingerstick blood have both been shown to be associated with added sugar intake, even after adjustment for meat intake. Researchers have attempted to isolate specific compounds in blood that are uniquely derived from dietary carbohydrates, such as direct endogenous carbohydrate sources (blood glucose) and specific non-essential amino acids (red blood cell alanine), and have seen strong correlations with added sugar intake. Recognized dietary confounders such as meat/animal products have been addressed using statistical adjustments and a dual-isotope analytical approach that invokes d15N as a correction factor. Further controlled feeding studies and epidemiological surveys complete with Institutional Review Board approval are needed to establish the sensitivity of d13C tissue assay as an objective biomarker for added sugar intake.

Objective:To describe relationships among baseline characteristics, engagement indicators and outcomes for rural participants enrolled in SIPsmartER, a behavioural intervention targeting sugar-sweetened beverage (SSB) intake.Design:A secondary data analysis. Bivariate analyses determined relationships among baseline characteristics (e.g. age, gender, race, education, income), engagement indicators (completion of 6-month health screening, class attendance, call completion) and SSB outcomes (SSB ounce reduction (i.e. US fluid ounces; 1 US fl. oz = 29·57 ml), reduced ≥12 ounces, achieved ≤8 ounce intake). Generalized linear models tested for significant effects of baseline characteristics on engagement indicators and of baseline characteristics and engagement indicators on SSB outcomes.Setting:South-west Virginia, USA, a rural, medically underserved region.Participants:Participants’ (n 155) mean age was 41 years; most were female (81 %), White (91 %) and earned ≤$US 20 000 per annum (61 %).Results:All final models were significant. Engagement models predicted 12–17 % of variance, with age being a significant predictor in all three models. SSB outcome models explained 5–70 % of variance. Number of classes attended was a significant predictor of SSB ounce reduction (β = −6·12, P < 0·01). Baseline SSB intake significantly predicted SSB ounce reduction (β = −0·90, P < 0·001) and achieved ≤8 ounce intake (β = 0·98, P < 0·05).Conclusions:The study identifies several participant baseline characteristics that may impact engagement in and outcomes from a community-based intervention targeting SSB intake. Findings suggest greater attendance of SIPsmartER classes is associated with greater reduction in overall SSB intake; yet engagement variables did not predict other outcomes. Findings will inform the future implementation of SIPsmartER and research studies of similar design and intent.

In the United States (U.S.), up to 40% of the entire food supply is lost or wasted, which contributes to negative environmental, economic, and social outcomes, with the majority of food waste occurring in the retail and consumer sectors of the food supply chain. Concurrently, 10.5% of U.S. households experienced food insecurity in 2019. Food insecurity rates are higher among individuals with low income and disproportionately affects ethically and racially diverse people living in poverty. Diverting excess food from retail and institutional settings to people in need is one method to reduce food waste and enhance individual, household, and community food security. This PhD dissertation describes four studies that explored food waste management in the U.S. among households with low-income and within U.S. food pantries, which are a critical component of the emergency food system. Study 1 consisted of a systematic review that examined differences in food waste attitudes and practices among consumers of across income levels. Study 2 was a quantitative research methods approach to examine food waste attitudes and practices of SNAP-eligible households (n=17). Study 3 was a cross-sectional quantitative approach to examine the impact of the novel coronavirus disease (COVID-19) pandemic on food waste management within U.S. food pantries (n=290). Study 4 assessed the feasibility of food pantries in Virginia (n=68) to implement marketing-mix and choice-architecture (MMCA) strategies within their food pantries given the changes they had to make in their food distribution models in response to COVID-19 physical distancing requirements. Collectively, the studies described in this PhD dissertation address knowledge gaps and may be used to inform policies and initiatives to increase food security and decrease food waste among households with low-income and organizations aimed at serving them. Doctor of Philosophy Food waste in the U.S. is a major challenge that impacts the environment, economy, and society. The largest amounts of food waste are generated within food retail and consumer household settings. Concurrently, food insecurity, which is defined as obtaining and having access to enough food for an adequate, nutritious diet, is also another complex food systems challenge in the U.S. Food insecurity rates are higher among people with low income and disproportionately affects ethnically and racially diverse people. The novel coronavirus (COVID-19) pandemic placed further pressure on these two food systems challenges. Diverting excess, wholesome food from retail and institutional settings to people with hunger is a preferred method to mitigate both food waste levels and food insecurity rates. The U.S. emergency food system, which is a network of food recovery organizations that aim to receive excess food from retail settings and redistribute the food to people with hunger, play a pivotal role in addressing these two challenges. This PhD dissertation described four studies that address these two challenges. While this dissertation examined food waste management, the first half of the dissertation explored this topic within households and the second half explored food waste management within food pantries, which are a critical component to the U.S. emergency food system. Study 1 utilized a systematic procedure to review previously published consumer food waste research. The purpose of this study was to examine differences in food waste attitudes and practices among consumers of various income levels. Study 2 used a survey to examine food waste attitudes and practices of households eligible for Supplemental Nutrition Assistance Program (SNAP) benefits. Study 3 explored the impact of COVID-19 physical distancing requirements on food waste management within U.S. food pantries. Lastly, study 4 explored the feasibility of food pantry staff/volunteers on implementing behavioral economics strategies, which are changes to a setting that can promote selection of a certain food item, within their food pantry. Collectively, the studies described in this PhD dissertation address knowledge gaps and may inform policies and initiatives to mutually increase food security and decrease food waste among households with low-income and organizations aimed at serving them.

The American Heart Association Life's Simple 7 (LS7), a metric that classifies cardiovascular health using modifiable risk factors, has been reported to be associated with cognitive function. However, the assessment of cognitive function in prior studies has been limited to relatively crude global measures. We hypothesized that greater LS7 scores at baseline are associated with less cognitive decline and lower incidence of dementia and Alzheimer's. Using data from the core Health and Retirement Study survey (2012-2018), 2013 Health Care and Nutrition Questionnaire and enhanced face-to-face interviews we will evaluate cognitive scores in relation to LS7. The participants included in this analysis (n = 2,753) are greater than 50 years of age without dementia. A global cognitive score was calculated using the combination scores on multiple cognitive tests that include measurements of semantic memory, quantitative reasoning, episodic memory, and executive functioning. Changes in cognitive scores are evaluated based on marginal effects after adjustments for confounding variables. Baseline LS7 scores (0-14) were calculated using information on smoking habits, body mass index, habitual diet, blood pressure, non, physical activity, and hemoglobin a1c. Results were analyzed using linear mixed models fit with random intercepts and the use of Huber-White variance estimates to analyze the results. Knots were introduced to explain non-linear change in respondents. Global cognitive scores in all respondents decreased more between baseline to year two and year two to year four than year four to year six (ΔB-2: -2.796, Δ2-4: -3.362 v. Δ4-6: -1.191). A one unit increase in LS7 score presented a protective effect and slowed the rate of decline by 0.11 unit in global cognitive scores between baseline and year two. The protective effect was lower between year two and four (0.07 units) and non-significant between year four and six. Black respondents did not respond similarly in models than white respondents. A one unit increase in LS7 score increased the rate of decline from baseline to year two and year two to year 4 in black respondents (p

Background: Poor beverage consumption habits pose significant health concerns. Delivering health behavior change interventions via social media is an emerging area of health research and may provide a promising way to minimize barriers such as cost, intervention delivery time, and access. However, there is limited research on online social support health behavior change delivered through Instagram. Objectives: 1) Assess the validity and reproducibility of the updated BEVQ-15, a beverage intake questionnaire; 2) Review the current literature on the availability and effectiveness of mobile phone interventions targeting sugar-sweetened beverage (SSB) consumption; and 3) Use the updated BEVQ-15 to conduct the Healthy Beverage Habits pilot study, which is an online social networking worksite intervention aimed at improving Healthy Beverage Index (HBI) scores. Methods: The Healthy Beverage Habits study was an online randomized controlled pilot trial with an 8-week intervention and a 4-week maintenance period delivered through Instagram. The materials were adapted from the in-person SIPsmartER SSB reduction intervention. Data analysis included RM-ANOVAs to test for differences in beverage intake between the Instagram intervention group and the E-mail control group. Results: For the Healthy Beverage Habits trial, no significant between group over time differences were found. However, the Instagram group demonstrated a significant reduction in total beverage kcal (mean difference±SE=-156±48; p=0.049), and increase in total HBI scores (mean difference±SE= 11.9±2.3; p=0.025) from baseline to the end of the maintenance period. No within group differences were demonstrated for the E-mail control group over the intervention or maintenance period. The study retention rate was 38%, with 39 participants initially enrolled and 15 participants completing all study visits through maintenance. Results for the validity and reproducibility of the updated BEVQ-15 and a review of the availability and effectiveness of mobile phone interventions targeting SSB consumption are presented. Conclusions: Mobile phone-delivered interventions may be a promising method for improving beverage intake quality. Technologically-based interventions targeting beverage consumption should consider utilizing multiple forms of mobile-phone contact methods. The preliminary findings from the Healthy Beverage Habits trial highlight the need for more rigorous studies that determine which technology and intervention components are most effective for mobile-delivered beverage consumption interventions. Doctor of Philosophy Background: Poor beverage consumption habits pose significant health concerns. Delivering health behavior change interventions via social media is an emerging area of health research and may provide a promising way to minimize barriers such as cost, intervention delivery time, and access. However, there is limited research on online social support health behavior change delivered through Instagram. Objectives: 1) Assess the validity and reproducibility of the updated BEVQ-15, a beverage intake questionnaire which estimates habitual average daily intake of 15 beverage categories as well as total sugar-sweetened beverages (SSB) and total beverages. 2) Review the current literature of the availability and effectiveness of mobile phone interventions targeting SSB consumption. 3) Use the updated BEVQ-15 to conduct The Healthy Beverage Habits pilot study, which is an online social networking worksite intervention aimed at improving Healthy Beverage Index (HBI) scores. Methods: The updated BEVQ-15 was assessed for agreement between the BEVQ-15 and dietary recalls. Researchers compared beverage intake between two BEVQ-15 administrations. The Healthy Beverage Habits pilot study was an 8-week intervention with a 4-week maintenance period. Data analysis included testing for differences in beverage intake between the Instagram group and the E-mail group. Results: The updated BEVQ-15 demonstrated moderate agreement between the BEVQ-15 and dietary recalls for total SSB intake and total beverage intake. All beverage variables were significantly correlated. For the systematic review, 11 of the 17 studies (65%) were successful in reducing SSB consumption through mobile phone delivery. The successful studies used multiple types of technology. There were no significant differences for any beverage variable between the Instagram and E-mail groups over time. However, for within group changes, Instagram significantly reduced total beverage kcals and improved total HBI score, while the E-mail group did not demonstrate any significant changes. Conclusions: Mobile phone-delivered interventions may be a promising method for improving beverage intake quality. Technologically-based interventions targeting beverage consumption should consider utilizing multiple forms of mobile-phone contact methods. The preliminary findings from the Healthy Beverage Habits pilot trial highlight the need for more rigorous studies that determine which technology and intervention components are most effective for mobile-delivered beverage consumption interventions.

Team sports frequently use inertial measurement units (IMU) fixed at the scapulae for the quantification of athlete performance. Similar IMU are used in clinical settings for gait analysis and jump testing but are located at the center of mass (COM). For clinical methods of jump and gait analysis to be translated to sports related IMU, an investigation of the validity of measure from the scapulae should be assessed. The objective of the current study was to translate clinical methods of gait analysis and jump assessment to commercially available trunk-mounted accelerometers. The current study created a gait program to analyze and compile the gait data. Following completion of the gait program, the IMU (STATSports APEX) was investigated for validity against ankle accelerometers. Once the validity was determined, an application study evaluated the relevance of collecting gait data during a NCAA D1 Women's Soccer season. Similarly, the trunk-mounted accelerometer was validated against force plates to assess countermovement jump height. The final study assessed how both jump height and gait variables changed due to game-related decline in performance. This study provides evidence that trunk-mounted accelerometers are a valid tool for assessing temporal gait variables (ICCRight = 0.95 and ICCLeft = 0.96), CMJ height (ICCJH = 0.90) and flight time (ICCFT = 0.88). A longitudinal analysis of gait showed that StepL, StrideL, kleg, and postural variables changed regularly in pre-post comparisons of performance. Postural variables had more changes towards the end of the season. Root mean squares (RMS) of accelerations and angular velocities had the highest correlations to High Speed Running (HSR). Fractal step and stride length (StepLα and StrideLα) had the strongest correlation to Total Distance (Rstep = -0.29 and Rstride = -0.29), or Tot Dist. When comparing gait and CMJ analyses to detect game-related changes in performance, CMJ was more descriptive of fatigue. In a proof-of-concept study, pre-post changes in CMJ immediately before and after a game had a moderate negative correlation (R = -0.57) to Tot Dist. When the protocol changed to assess the differences between the days before and after the game, the correlation weakened to R = -0.27. Spatiotemporal and spring mass variables did not change, whereas postural variables appeared to improve. The current study has provided evidence that running gait could be used as an athlete monitoring technique, however more data needs to be collected to understand how running gait variables change with team-sports related fatigue. Doctor of Philosophy Trunk-mounted accelerometers have become widely popular in team sports such as soccer. Prior to their use in team sports, accelerometers were used in clinical settings to assess gait, and in some cases jump performance. Different from the trunk-mounted accelerometers, the clinical accelerometers were fixed on the lower back to approximate the position of the center of mass. Consequently, the methods to assess gait and jump performance using an accelerometer assume that the accelerometer is fixed on the lower back. For these methods to be translated to trunk-mounted accelerometers, the validity of the methods needs to be established. This paper investigated the validity of trunk-mounted accelerometers in evaluating gait and countermovement jump height. A season-long assessment of gait provided insight into the utility of monitoring gait in team sports athletes. Lastly, a comparison of gait to countermovement jump analyses helped to elucidate what the changes in gait indicate, and how team staff may effectively utilize that information. The trunk-mounted accelerometer (STATSports APEX) was valid in estimating gait and jump variables. The seasonal analysis showed that gait variables changed frequently throughout the season. The main variables affected described the step and stride length of the individual, the stiffness of the leg, and variables representing the variability in trunk movements associated with running. The variability in trunk movements were more perturbed towards the end of the season when players may be beginning to tire from continuous training and competition. In the final study, the jump analysis proved to be more representative of the player's state of fatigue than gait. The gait variables appeared to improve after exhaustive exercise. The apparent improvement in gait variables could indicate that the players recovered quickly from the exhaustive exercise, or that gait variables respond in a more complex fashion than jump variables. More data needs to be collected on how gait changes due to game-related fatigue in soccer players to better understand its effect on neuromuscular performance.

Colorectal cancer is the third leading cause of cancer death in the USA, yet is highly preventable and detectable at an early stage through screening. Virginia Cooperative Extension (VCE) implemented a worksite colon cancer awareness program to increase colorectal cancer screening rates and preventive lifestyle behaviors among its employees. The Colon Cancer–Free Zone program is designed using best practice principles of worksite health programs and includes information sessions covering the topics of colorectal cancer, screening guidelines, insurance coverage, and preventive lifestyle behaviors. It is conducted in a campaign format that includes a strategic communication strategy targeting relevant screening barriers and facilitators, peer champions, and incentives. The program was implemented with VCE employees statewide utilizing a web-based system for the information sessions, and resulted in broad participation, a significant increase in screening self-efficacy (4.15 ± 0.64 vs 3.81 ± 0.76, ρ = 0.006), changes in diet and physical activity (50% and 40% of participants, respectively), and a 20.6% increase in the employee colorectal cancer screening rate. A Colon Cancer–Free Zone toolkit was developed for use by Extension Agents to implement the program at worksites in their service communities. Electronic supplementary material The online version of this article (10.1007/s13187-019-01569-4) contains supplementary material, which is available to authorized users.

Angiotensin II (ANG II)-induced skeletal muscle wasting is characterized by activation of the ubiquitin-proteasome system. However, the potential involvement of proteolytic system macroautophagy/autophagy in this wasting process remains elusive. Autophagy is precisely regulated to maintain cell survival and homeostasis; thus its dysregulation (i.e., overactivation or persistent suppression) could lead to detrimental outcomes in skeletal muscle. Here we show that infusion of ANG II for 7 days in male FVB mice suppressed autophagy in skeletal muscle. ANG II blunted microtubule-associated protein 1 light chain 3B (LC3B)-I-to-LC3B-II conversion (an autophagosome marker), increased p62/SQSTM1 (an autophagy cargo receptor) protein expression, and decreased the number of autophagic vacuoles. ANG II inhibited UNC-51-like kinase 1 via inhibition of 5′-AMP-activated kinase and activation of mechanistic target of rapamycin complex 1, leading to reduced phosphorylation of beclin-1Ser14 and Autophagy-related protein 14Ser29, suggesting that ANG II impairs autophagosome formation in skeletal muscle. In line with ANG II-mediated suppression of autophagy, ANG II promoted accumulation of abnormal/damaged mitochondria, characterized by swelling and disorganized cristae and matrix dissolution, with associated increase in PTEN-induced kinase 1 protein expression. ANG II also reduced mitochondrial respiration, indicative of mitochondrial dysfunction. Together, these results demonstrate that ANG II reduces autophagic activity and disrupts mitochondrial ultrastructure and function, likely contributing to skeletal muscle wasting. Therefore, strategies that activate autophagy in skeletal muscle have the potential to prevent or blunt ANG II-induced skeletal muscle wasting in chronic diseases. NEW & NOTEWORTHY Our study identified a novel mechanism whereby angiotensin II (ANG II) impairs mitochondrial energy metabolism in skeletal muscle. ANG II suppressed autophagosome formation by inhibiting the UNC-51-like kinase 1(ULK1)-beclin-1 axis, resulting in accumulation of abnormal/damaged and dysfunctional mitochondria and reduced mitochondrial respiratory capacity. Therapeutic strategies that activate the ULK1-beclin-1 axis have the potential to delay or reverse skeletal muscle wasting in chronic diseases characterized by increased systemic ANG II levels.

High ultra-processed food (UPF) consumption is associated with weight gain, obesity, metabolic syndrome, cardiovascular disease, all-cause mortality, and type 2 diabetes. "Food agency" refers to an individual's food preparation attitudes and capabilities within a given social and physical food environment. Although there is an association between the ability to prepare certain dishes and higher cooking frequency with lower UPF intake, it is still unclear if cooking and food agency influences UPF consumption due to the lack of research in this area. To date, studies of food agency have not included detailed dietary analyses. This cross-sectional study included 40 adults (73% female, aged 39±15 years, body mass index [BMI] 25.8±4.9 kg/m2) and aimed to determine the relationship between food agency and UPF intake. Food agency scores were calculated using the validated Cooking and Food Provisioning Action Scale (CAFPAS). Cooking behaviors and frequency were also collected. Dietary intake was assessed using three 24-hour dietary recalls with each item categorized by degree of processing using the NOVA classification system. Analyses included one-way ANOVAs between CAFPAS score quartiles [Q] and Pearson correlations between CAFPAS, cooking behaviors, and dietary intake. UPF intake was calculated in percent total calories and percent total grams to account for artificial sweeteners and other low calorie products that may be classified as UPFs. Low food agency was associated with a 14% higher UPF intake (% total calories) compared to high food agency (P=0.03). There were no differences between CAFPAS quartiles and UPF intake in % total grams. When age was divided into three categories, UPF intake was significantly lower in older adults (60+ years) compared to adults aged 19-39 years and 40-59 years by 26.4% and 29.1% respectively (P=0.02). When analyzed using bivariate analysis, lunch (r=-0.482, P=0.002) and dinner (r=-0.385, P=0.014) cooking frequency, frequency of cooking a meal from scratch or fresh ingredients (r=-0.320, P=0.044), and CAFPAS self-efficacy (r= -0.369; P= .019) were negatively correlated with UPF intake (% total kcal). These findings provide support for the possibility of reducing UPF intake through cooking interventions aimed at improving components of food agency such as problem solving, skills to create a meal from what is available, and cooking confidence. Future studies should include a larger, heterogeneous population to provide more insight into dietary differences between levels of food agency and expand the diversity of research in this area. Master of Science High ultra-processed food (UPF) consumption is linked to weight gain, obesity, metabolic syndrome, cardiovascular disease, all-cause mortality, and type 2 diabetes. "Food agency" refers to an individual's food preparation attitudes and capabilities within a given social and physical food environment. Although there is a link between the ability to prepare certain dishes and higher cooking frequency with lower UPF intake, it is still unclear if cooking and food agency influences UPF consumption due to the lack of research in this area. To date, studies of food agency have not included detailed dietary analyses. This study aimed to determine the relationship between food agency and UPF intake. Food agency scores were calculated using the validated Cooking and Food Provisioning Action Scale (CAFPAS). Cooking frequency and behaviors were also collected. Dietary intake was assessed using three 24-hour dietary recalls with each item categorized by degree of processing using the NOVA classification system. Participants with low food agency had higher UPF intake in percent total calories compared to those with high food agency. There were no differences between food agency and UPF intake in percent total grams. Adults aged 60 and over consumed less UPF compared to adults below the age of 60. Participants with higher food agency were more likely to make a meal from scratch or fresh ingredients. As frequency of cooking a meal from scratch or fresh ingredients, cooking dinner or lunch increased, and CAFPAS self-efficacy scores increased percent total calories from UPFs decreased. These findings provide support for the possibility of reducing UPF intake through cooking interventions aimed at improving components of food agency such as problem solving, skills to create a meal from what is available, and cooking confidence. Future studies should include a larger, heterogeneous population to provide more insight into dietary differences between levels of food agency and expand the diversity of research in this area.

American football is one of the most popular sports in the United States. However, in comparison to other mainstream sports such as soccer and rugby, there is limited literature using scientific principles and theory to examine the most appropriate ways to monitor the sport. This serves as a barrier to American football practitioners in their development and implementation of evidence-based sport preparation programs. Therefore, the primary aim of this line of research (i.e., dissertation) is to illustrate the efficacy of commonly used athlete monitoring tools within the sport of American collegiate football, while proposing a systematic framework to guide the development of an athlete monitoring program. This aim was achieved through a series of studies with the following objectives: 1) to quantify the physical demands of American collegiate football practice by creating physiological movement profiles through the use of integrated microtechnology metrics and heart rate indices, 2) to determine the positional differences in the physical practice demands of American collegiate football athletes, 3) to examine which integrated microtechnology metrics might be used to most efficiently monitor the training load of American collegiate football athletes, 4) to demonstrate the suitability of using the countermovement jump (CMJ) to assess training adaptations in American collegiate football athletes through examining weekly changes in CMJ performance over the course of two 4-week periodized training blocks (8 weeks total), and 5) to examine the effect of acute fatigue on CMJ performance in American football athletes. The first study from this line of research quantified the physical demands of American collegiate football by position groups and found significant differences in both running based and non-running based training load metrics. In addition, the first study utilized a principal component analysis to determine 5 'principal' components that explain approximately 81% of the variance within the data. The second study utilized a univariate analysis and found significant changes in CMJ performance due the effect of time with significant improvements in CMJ 'strategy' variables over the training period. Finally, the third study used effects sizes to illustrate a larger magnitude of change in CMJ 'strategy' variables than CMJ 'output' variables due to effect of acute fatigue. Results from studies 2 and 3 suggest the importance of monitoring CMJ strategy variables when monitoring training adaptations and fatigue in American collegiate football athletes. This line of research provides practitioners with a systematic framework through which they can develop and implement evidence-based sport preparation programs within their own organizational context. In addition, this line of research provides practitioners with recommendations for which metrics to monitor when tracking training load in American collegiate football using integrated microtechnology. Finally, this line of research demonstrates how to assess training adaptations and fatigue using the CMJ within the sport of American collegiate football, while providing an empirical base through which the selection of CMJ variables can take place. Collectively, this line of research uses scientific principles and theory to extend the current literature in American collegiate football, while providing practitioners with a guide to athlete monitoring within the sport. Doctor of Philosophy American football is one the most popular sports in the United States. Despite its popularity, there is limited research using scientific principles and theories to examine ways to most effectively monitor the sport. Broadly, athlete monitoring refers to the process of providing informational feedback from the athlete to practitioners. This allows practitioners to make decisions informed by data. Therefore, this line of research (i.e. dissertation) aimed to use a variety of commonly used athlete monitoring tools to monitor American collegiate football athletes, while proposing a framework to guide in the development of an athlete monitoring program. This line of research consisted of a series of 3 studies. In study #1, it was found that integrated microtechnology units and heart rate sensors could be used to determine the physical demands of American collegiate football practice, as well as differences in the physical demands of practice by position group. In addition, a set of 5 training load constructs were found through which training load in American collegiate football athletes may be appropriately monitored. In study #2, it was found that countermovement jump (CMJ) strategy variables indicating how the jump occurred may provide more insight into strength and power training adaptations than CMJ output variables that indicate what occurred as a result of the jump in this highly trained athletic population. Finally, in study #3, it was found that CMJ strategy variables may be more sensitive to acute fatigue from a football-specific training session than CMJ output variables in American collegiate football athletes. Collectively, this research suggest that integrated microtechnology units, heart rate sensors, and the CMJ using a force testing platform may be used to monitor American collegiate football athletes. Moreover, this research suggests which variables to utilize when monitoring this population using these tools through the proposed athlete monitoring framework.

There is an association between the human microbiome and disease. Gut microbes metabolize dietary sources to release trimethylamine (TMA). TMA is absorbed and then oxidized by flavin monooxygenase 3 (FMO3) to form trimethylamine N-oxide (TMAO). Elevated TMAO is associated with increased risk of cardiovascular disease and type 2 diabetes; however, the causal nature is unclear. There is also limited evidence supporting the efficacy of strategies to reduce accumulation of TMAO. Therefore, the purpose of these studies is to determine the effects of increases in TMAO on cardiometabolic health. In study 1, healthy sedentary and endurance trained males consumed a high fat diet. Blood samples were obtained in a fasted state and every hour during a 4-hour high fat challenge. We hypothesized sedentary individuals would produce higher TMAO concentrations. In study 2, healthy sedentary individuals consumed an acute 1000 mg dose of choline (CHOL) and placebo (PLC). Fasted blood samples were collected, flow-mediated dilation (FMD) and oral glucose tolerance (OGT) were measured. In study 3, healthy sedentary individuals consumed 4-wks of CHOL and PLC. Fasted blood samples were collected, FMD and OGT were measured. We hypothesized acute and 4-wk choline supplementation would impair FMD and OGT. In study 1, neither fasting (1.49± 1.2 µM vs. 2.25 ± 1.4 µM, p>0.05) or postprandial TMAO changed significantly with the HFD in sedentary or endurance trained individuals even with the endurance group consuming more TMA dietary precursors. Study 2 found increased plasma TMAO concentrations after choline supplementation on day 1(PLC; 4.14 ± 2.6 μM vs. CHOL; 23.6 ± 33.8 μM, p=0.018) and day 2 (PLC; 5.13±4.9 μM vs. CHOL; 32.6±37.5 μM, p=0.082) however, there were no differences in OGT or FMD. Study 3 found no differences in FMD or OGT following 4-wks of choline consumption. In summary, there were no differences between sedentary and endurance trained individuals fasting or post-prandial TMAO. There was also no effect on acute or 4-wk supplementation of choline on FMD and OGT. More research is needed to understand effects of elevated TMAO on cardiometabolic health. Doctor of Philosophy For years, research has been performed to identify the health effects of eating large amounts of red meat on cardiovascular disease (CVD). Consuming red meat, fish, poultry and eggs increases a substance created during digestion and metabolism, called trimethylamine N-oxide (TMAO). Elevated TMAO has been associated with increased risk of CVD and type 2 diabetes but the direct causes are unknown. The purpose of these studies is to determine the effects of increases in TMAO on health in humans. Study 1 included healthy, sedentary and endurance trained males who consumed a high fat diet. Blood samples were collected to measure TMAO before and after a high fat meal. Study 2 included healthy, sedentary males and females who consumed 2 days of 1000 mg of choline, which is commonly found in red meat fish and eggs, and a placebo (carbohydrate) after subjects completed a series of tests to evaluate health. Study three included healthy, sedentary males and females who consumed 4-weeks of 1000 mg of choline per day and a placebo (carbohydrate). Following supplementation subjects underwent a series of tests to assess health. Overall, there were no differences found between sedentary and endurance trained individuals. Acute and 4-week supplementation of choline did not affect measures of blood sugar or blood vessel function.

There is a critical gap in translating scientific discoveries to public health benefit. For example, despite a multitude of efficacious physical activity interventions, only one in four adults in the United States meets the Physical Activity Guidelines for Americans. To bridge the research-practice gap, Dissemination and Implementation (DandI) Science has emerged as the study of how evidence-based interventions, programs, and policies are integrated in typical settings. Recent research illustrates barriers to conducting DandI Science and the need for methods that open the black box of implementation. Therefore, the overarching goal of this dissertation was to explore novel approaches for advancing DandI Science. This exploration is presented in three manuscripts and one report. The first manuscript presents a pragmatic, observational study applying the RE-AIM (Reach, Effectiveness, Adoption, Implementation, Maintenance) Framework to evaluate a perinatal health fair. Results include that the health fair reached 42 attendees and that 23 educators and seven organizations hosted booths and educational sessions. Mom Expo required 292 implementation hours with 71% of those hours devoted to building relationships. We generated 30 actionable strategies for implementing a health fair. The health fair developed into a non-profit organization, and the participatory approach used can be replicated in other communities to establish connections between local women, educators, and researchers. The second manuscript reports a one-year autoethnography (i.e., first-person narrative) of a perinatal health integrated research practice partnership (IRPP). Findings include three themes: (1) permeable work boundaries, (2) blind spots toward philosophical underpinnings of paradigms, and (3) maladaptive behaviors seemingly reinforced by the research culture. We concluded that autoethnography is an effective novel method to leverage researcher situatedness and capture implementation contexts, processes, and outcomes. The third manuscript presents the longitudinal pilot test of FUEL (focus, unplug, exercise, love), a one-on-one coaching program to promote human flourishing among DandI researchers. Results include that the coach spent 12.96+2.82 hours per participant (N= 16) implementing individually-tailored sessions, and that participants reported multiple, sustained benefits related to productivity, happiness, and health. We concluded that the program is a feasible, well-received approach with preliminary positive effects. Future work is needed to investigate physiological or performance outcomes and, ultimately, impact on DandI. The final report is a literature review and critical analysis of phenomenology within behavioral and community health research. Conclusions include that (1) physical activity is rooted in a scientific paradigm that prioritizes quantifiable mechanism over personal meaning, and (2) phenomenology, as a complement to basic science, is a compelling method, paradigm, and practice to improve research translation. Based on this research, I conclude that three pathways for advancing DandI Science are methods that capture first-person meaning, paradigms that incorporate phenomenological human experience as an essential dimension of health research, and practices that fuel researchers' capacity for generating transformative work. In all three pathways, the heart of elevating DandI Science is to embrace process, person, and presence. Doctor of Philosophy Scientific evidence does not automatically translate to real-world behavior change. For example, despite considerable research about the health benefits of physical activity, only one in four American adults meets the national physical activity recommendations. To bridge the research-practice gap, Dissemination and Implementation (DandI) Science has emerged as the study of how scientific findings are integrated in typical settings such as schools and communities. Recent research illustrates multiple barriers to DandI Science and a need for methods that capture hard-to-measure, chaotic implementation processes and outcomes. Therefore, the overarching goal of this dissertation was to explore novel approaches to DandI Science and bridging the research-practice gap. This exploration is presented in three manuscripts and one report. The first manuscript describes a perinatal health fair intended to connect local parents to community resources. The second manuscript is a 12-month autoethnography (i.e., first-person narrative) about the culture of DandI Science, including the role, impact, and practices of researchers themselves. The third manuscript presents the development and preliminary testing of FUEL (focus, unplug, exercise, love), a one-on-one coaching program for DandI researchers. The final report includes the history of randomized controlled trials as the gold standard for physical activity research, as well as critical analysis of using phenomenology to reduce the research-practice gap. Findings from the first manuscript suggest that (1) authentic relationship building was the key to launching a perinatal health fair that developed into a non-profit organization and (2) the RE-AIM (Reach, Effectiveness, Adoption, Implementation, Maintenance) Framework is a user-friendly DandI tool for implementing and evaluating a health fair. Findings from the second manuscript include descriptions of (1) blind spots within the DandI Science culture, (2) potential of autoethnography as a novel DandI method, and (3) strategies to optimize DandI researchers' capacity to thrive amid challenges. Findings from the third manuscript suggest that the FUEL coaching program is a promising and feasible approach to support researchers in leading "a more productive, healthier, and happier life," as one participant wrote. Future research on the program is needed to evaluate causation and whether organizations would adopt it. Conclusions in the final report include that (1) the applicability of physical activity research to daily life may be limited by deeply held scientific ideologies and (2) phenomenology, as the study of human meaning, may facilitate the translation of research to real-world behavior change. Based on the research presented in this dissertation, three pathways for advancing DandI Science are methods for how we conduct research studies, paradigms for how we collectively approach health science, and practices for how we manage our energy and awareness. In all three pathways, the heart of elevating DandI Science is to embrace process, person, and presence.

Sex difference in adiposity has long been recognized but the mechanism remains incompletely understood. Previous studies suggested that adiposity was regulated by autophagy in response to energy status change. Here, we show that the energy sensor Sirt1 mediates sex difference in adiposity by regulating autophagy and adipogenesis in partnership with estrogen receptor α (ERα). Autophagy and adipogenesis were suppressed by Sirt1 activation or overexpression, which was associated with reduced sex difference in adiposity. Mechanistically, Sirt1 deacetylated and activated AKT and STAT3, resulting in suppression of autophagy and adipogenesis via mTOR-ULK1 and p55 cascades. ERα induced Sirt1 expression and inhibited autophagy in adipocytes, while silencing Sirt1 reversed the effects of ERα on autophagy and promoted adipogenesis. Moreover, Sirt1 deacetylated ERα, which constituted a positive feedback loop in the regulation of autophagy and adiposity. Our results revealed a new mechanism of Sirt1 regulating autophagy in adipocytes and shed light on sex difference in adiposity. American Heart Association GrantAmerican Heart Association [18TPA34230082]; USDA National Institute of Food and Agriculture Grant [1020373] This work was supported in part by the American Heart Association Grant (18TPA34230082 to Z.C.), and the USDA National Institute of Food and Agriculture Grant (1020373 to Z.C).

BackgroundThere is a need to unpack the empirical, practical, and personal challenges within participatory approaches advocated to optimize implementation. The unpredictable, chaotic nature of participatory approaches complicates application of implementation theories, methods, and strategies which do not address researchers’ situatedness within participatory processes. As an implementation scientist, addressing one’s own situatedness through critical reflection is important to unearth how conscious and unconscious approaches, including ontological and epistemological underpinnings, influence the participatory context, process, and outcomes. Therefore, the aim of this exploratory work is to investigate the heretofore blind spot toward the lived experience of implementation researchers within the participatory process.MethodsWe developed an integrated research-practice partnership (IRPP) to inform the implementation of a gestational weight gain (GWG) control program. Within this IRPP, one investigator conducted a 12-month autoethnography. Data collection and triangulation included field notes, cultural artifacts, and systematic timeline tracking. Data analysis included ethnographic-theoretical dialogue and restorying to synthesize key events and epiphanies into a narrative.ResultsAnalysis revealed the unpredicted evolution of the GWG program into a maternal health fair and three themes within the researchers’ lived experience: (1) permeable work boundaries, (2) individual and collective blind spots toward the ontological and epistemological underpinnings of implementation paradigms, and (3) maladaptive behaviors seemingly reinforced by the research culture. These themes contributed to the chaos of implementation and to researchers’ experience of inadequate recovery from cognitive, emotional, and practical demands. These themes also demonstrated the importance of contextual factors, subjectivity, and value-based judgments within implementation research.ConclusionBuilding on extant qualitative research guidelines, we suggest that researchers anchor their approach to implementation in reflexivity, intentionally and iteratively reflecting on their own situatedness. Through this autoethnography, we have elucidated several strategies based on critical reflection including examining philosophical underpinnings of research, adopting restorative practices that align with one’s values, and embracing personal presence as a foundation of scientific productivity. Within the predominant (post-) positivism paradigms, autoethnography may be criticized as unscientifically subjective or self-indulgent. However, this work demonstrates that autoethnography is a vehicle for third-person observation and first-person critical reflection that is transformative in understanding and optimizing implementation contexts, processes, and outcomes.

This commentary describes insights from Star Trek's fictional television series to understand how state and non-state actors address conflicts of interest (COIs) through global nutrition governance. I examine the findings of Ralston and colleagues for 44 state and non-state actors who responded to the World Health Organization's (WHO's) consultation for a COI risk-assessment tool, developed for member states to engage effectively with non-state actors to address malnutrition in all forms. Star Trek reveals that actor engagement is inevitable in a shared universe. The Prime Directive is a non-interference principle reflecting a moral commitment to reduce harm, respect autonomy and protect rights. Engagement principles are relevant to all actors who influence nutrition policies and programs, and must be held accountable when their actions undermine healthy and sustainable food systems. Certain actors use COI to justify non-engagement with commercial actors yet competing interests, biases, corruption and regulatory capture are distinct challenges to manage. Finally, Star Trek's characters serve as allegories to understand actors' motives and actions to promote healthy and sustainable food systems. Unlike non-state actors, states are legally required to achieve their commitments and targets in the United Nations' (UN) Decade of Action on Nutrition (2016-2025) and Sustainable Development Goals (SDGs) 2030 Agenda. Published (Publication status)

For years, heart rate (HR) kinetics have been used as an indicator of training status and fatigue. Slowed kinetics indicate poor fitness and/or fatigue. In this study it was determined that HR kinetics can be reliably estimated during modeled, quasi-binary, and dynamic exercise using cross-correlation analysis of HR and external work rate. Heart rate and running speed were cross-correlated yielding a cross-correlation function (CCF) and analyzed for its peak (CCFmax) and time delay (CCFlag). Modeled exercise data where the time constants for HR (τ) increased from 2-120 sec. yielded linear decreases in CCFmax (r2 = 0.9949) and linear increases in CCFlag (r2=0.9996). A strong linear relationship existed between CCFmax and CCFlag (r2=0.9989). Steady-state exercise data produced strong relationships between the calculated τ and CCFmax (r2=0.8736) and CCFlag (r2=0.9061), and CCFmax and CCFlag showed a positive relationship (r2=0.7753). CCFmax between repeated sprint trials (R2=0.9123) and super set trails (R2=0.9227) were very similar. These results suggest good repeatability for both quasi-binary activity and random activity. To assess validity, CCFmax values during the sprint trials were compared to two standard field tests of fitness (Beep and Man U tests). There were strong relationships between CCFmax and distances covered during the Beep (r2=0.7911) and Mann U tests (r2=0.7770). Lastly, the applicability of the CCF method was applied to dynamic exercise, using data collected from competitive soccer matches. For the field players, significant reductions in CCFmax occurred during the first and second periods of the match. There was a significant relationship between the total distance covered during the match and the decline in CCFmax (r=-0.4297, p

Obesity is associated with deficits in cognitive function, particularly within the domain of executive function (EF). EF refers to higher order cognitive processes that regulate our ability to sustain attention, inhibit subconscious tendencies, remember and manipulate information for immediate use, and remain cognitively flexible. Deficits in EF in overweight and obese individuals may impact the success of weight loss and maintenance efforts. Therefore, understanding the biological links between obesity and EF, as well as the ability to reverse EF deficits with weight loss, is imperative. The first study aimed to determine the effect of weight loss in overweight and obese, middle-aged and older adults on serum brain-derived neurotrophic fact (BDNF), S100 calcium binding protein B (S100B), and glial fibrillary acidic protein (GFAP). Serum samples (n=21; 50-75 years, BMI 25-40 kg/m2) were pooled from two prior weight loss studies. Fasting blood measurements were taken before and after 8- or 12-weeks of hypocaloric diet-induced weight loss (1200 or 1500 kcal/d). Body Mass Index (BMI), body weight, waist circumference, and percent body fat (All p

The Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework was used to evaluate the impact of the Virginia Department of Education's (VDOE) implementation of the USDA's Team Nutrition training program on Virginia elementary schools from 2017-2020. The VDOE provided training for school nutrition directors (SNDs) at workshops in June 2018 and September 2019 and continued technical assistance to help SNDs improve the school food environment within their divisions. SNDs' perceived support, self-efficacy, and intention to implement changes to the school food environment was measured through pre- and post-training surveys and quarterly check-ins. Implementation was measured using lunchroom environment observations and strategic plans created at the workshops. SNDs representing 83.9% of Virginia elementary students (512,953 students) in 111 divisions attended at least one of the workshops. Within effectiveness, perceived support from cafeteria staff was highest among all stakeholders for both the 2018 and 2019 trainings, though it significantly decreased from before to after the training for both workshops. Average perceived support did not significantly change over time. Self-efficacy was significantly higher following the 2018 training workshop only. The intervention was adopted by 84% of divisions and both intention to implement and actual implementation were highest for strategies related to Farm to School, a VDOE priority. Effectiveness and implementation are expected to improve as it takes up to years for changes to occur in schools. Despite RE-AIM not being part of the initial evaluation plan, 62% of indicators were assessed. With some adaptations to improve effectiveness, the intervention should continue and be expanded. Master of Science Interventions to improve the dietary habits of children are often conducted in schools, as most children attend school and children consume up to two meals while at school. USDA's Team Nutrition initiative trains school nutrition staff to serve healthy school meals and provides materials to implement nutrition education for students. The Virginia Department of Education implemented the Team Nutrition program statewide through two training workshops for school nutrition directors (SNDs), June 2018 and September 2019, and technical assistance between the trainings. This study examined the process of implementing the intervention, as well as the outcomes. Overall, 84% of Virginia elementary students were represented by SNDs at the training workshops. SNDs were most likely to intend to implement and implement changes in their divisions related to Farm to School programs and least likely change their culinary skills trainings for staff. It can take up to two years after a training for changes to be implemented in schools, therefore it may be too early to know the full effect of the training. The results showed that the intervention was widely adopted, with 84.1% of divisions in Virginia attending at least one of the training workshops. Although participation was high in-person, less SNDs participated in the intervention activities outside of the trainings. Overall, the intervention was successful in reaching many students and had a small positive effect. With some adaptations to improve the effectiveness, the intervention should continue to be implemented and expanded to the rest of the state.

Ovarian cancer is considered to be one of the deadliest gynecological diseases. Over 21,000 women are expected to be diagnosed with this fatal disease in 2020 alone. Obesity, but more specifically a high waist-to-hip ratio, is indicative of abdominal obesity and has been correlated with increased risk of ovarian cancer. How abdominal obesity contributes to this increased risk has not been clearly delineated but much of the current research has been focused on the role of adipocytes. However, in addition to the adipocytes, abdominal white adipose tissue contains the stromal vascular fraction (SVF) which includes stem and progenitor cell populations, immune cells, and fibroblasts. Since the SVF can also be recruited by the cancer cells, we investigated how obesity affects the survival and metastatic potential of cancer cells by investigating changes in the expression of genes that contribute to survival, proliferation, migration, adherence, and invasion. We used culture conditions that mimic the non-permissive peritoneal environment. Cancer related genes, such as Dkc1, Ccnd2, Lig4, and Snai2, were upregulated when adipose derived stem cells (ADSC) were added into MOSE-LTICv spheroids. It was found that peritoneal serous fluid (PSF) from obese mice significantly increased migration of MOSE-LTICv (Serum vs PSF, 517.8 vs 1158.6). These studies brought new knowledge into the field of obesity and ovarian cancer risk and provided direction for future studies involving potential cellular and molecular targets for ovarian cancer diagnosis and treatment. Master of Science Ovarian cancer affects many women in the United States. Obesity or more specifically, carrying more weight around the waist, can affect a woman's risk of developing ovarian cancer. Abdominal fat needs to be researched to see if abdominal obesity can affect ovarian cancer on the cellular level. Researchers have looked into how fat cells, known as adipocytes, can affect the progression of ovarian cancer, but more research needs to be done on the contributions of other cells found within adipose tissue. Other cells in abdominal fat include cells such as immune cells, stem and progenitor cells and fibroblasts. We have explored how adipose stem cells from obese mice affect the DNA or "the blueprints" of the cells, survival, and progression of mouse ovarian cancer cells. We found that when adipose stem cells are combined with ovarian cancer cells the expression of certain genes or particular "blueprints" increased. The genes whose expression increased included Dkc1, Ccnd2, Lig4, and Snai2 and when deregulated can cause ovarian cancer cells to become more aggressive. The abdominal fluid from obese mice was found to increase migration of ovarian cancer cells which simulates an increase in metastatic potential. This information has given new insight into the obesity and ovarian cancer relationship.

Much research has shown that regular physical activity influences both physiological and psychological health. More specifically, physical activity positively impacts wellbeing, across physical, emotional and social wellness. Furthermore, group-based physical activity has been linked to increasing adherence to an exercise protocol as well as improving social connectedness. For college students, collegiate recreation facilities are the main providers of physical activity services and programming for the campus population to utilize. In the collegiate recreation literature, several university studies have pointed out that students who participate in their recreational facilities and programs have benefited from positive feelings of wellbeing. The purpose of this study was to investigate the impact that Virginia Tech Recreational Sports' group-based fitness programming has on Virginia Tech students' overall wellbeing. Wellbeing was defined by six dimensions: career (or sense of purpose), social, financial, community, physical and emotional. This study used a concurrent explanatory mixed methods design, composed of a brief cross-sectional survey and focus groups. This work simultaneously contributes to collegiate recreation by providing evidence that students who utilize facilities and programs have higher perceptions of wellbeing as well as highlighting suggested strategies for improvement including resource limitations and communication barriers. Finally, in terms of group fitness and wellbeing, this thesis provides preliminary evidence that those who participate in collegiate group exercise programs have higher perceptions of physical and social wellbeing. Master of Science Regular physical activity influences both physical and mental health. More specifically, physical activity positively impacts wellbeing, across physical, emotional and social wellness. Furthermore, group-based physical activity has been linked to increasing adherence to an exercise protocol as well as improving social connectedness. For college students, collegiate recreation facilities are the main providers of physical activity services and programming for the campus population to utilize. In the collegiate recreation literature, several university studies have pointed out that students who participate in their recreational facilities and programs have benefited from positive feelings of wellbeing. The purpose of this study was to investigate the impact that Virginia Tech Recreational Sports' group-based fitness programming has on Virginia Tech students' overall wellbeing. Wellbeing was defined by six dimensions: career (or sense of purpose), social, financial, community, physical and emotional. This study used a concurrent explanatory mixed methods design, composed of a brief cross-sectional survey and focus groups. This work simultaneously contributes to collegiate recreation by providing evidence that students who utilize facilities and programs have high perceptions of wellbeing as well as highlighting suggested strategies for improvement related to access, programs, variety and communication. Finally, in terms of group fitness and wellbeing, this thesis provides preliminary evidence that those who participate in collegiate group exercise programs have higher perceptions of physical and social wellbeing.

Celebrity endorsement is a global billion-dollar business used by food, beverage and restaurant companies to influence dietary behaviors that may contribute to overweight and obesity. Evidence suggests that existing government policies and industry self-regulatory programs in the United States (US) do not adequately protect young Americans from celebrity endorsement that promotes energy-dense and nutrient-poor food and beverage products. This PhD dissertation describes four studies that examine the nature, extent, and perceived influence of celebrity endorsement used by business firms and non-profit organizations to promote branded food and beverage products or branded campaigns to American children, adolescents and millennial young adults to inform future research and policies to promote healthy food environments. Study one created a database of US celebrities (n=732) involved with food and beverage group, brand or product endorsements (1990-2017) to examine celebrity profiles and their endorsement relationships, company partnerships, the nutritional profile of products and brands. Evidence was analyzed using Python version 3.5.5 and SPSS version 24. Study two examined celebrities associated with the Partnership for a Healthier America's Fruits and Veggies (FNV) Campaign (2015-2016). Evidence was analyzed using data visualization tools supported by Python and SPSS, and results informed the FNV Campaign. Study three used Q methodology to explore the views of Millennials, born 1981-1994 (n=40) at Virginia Tech about celebrity endorsement of food and beverage products and brands. Participants sorted 48 celebrity images depicting brand and product endorsements on a normal distribution (+4 to -4) based on celebrity trustworthiness and complete a post Q-sort questionnaire. Data analysis used PQMethod 2.35 statistical software program, centroid factor analysis, and qualitative interpretation of unique factor arrays for sorters. Study four identified relevant evidence about US celebrity endorsement of food and beverage products (2000-2019), used an accountability framework to evaluate the adequacy of accountability structures, and suggested future policies and action needed for diverse stakeholders to use celebrity endorsement to promote healthy food environments. Future research should explore the trend of celebrity endorsement used in the US marketplace and examine the influence of this marketing strategy on young people's food preferences and choices based on more representative experimental research. Doctor of Philosophy Celebrity endorsement is a marketing strategy that represents a person who uses public recognition to promote the sales, use or consumption of a brand, product, or service. Celebrity endorsement has been used by food, beverage and restaurant companies to influence eating behaviors that may contribute to overweight and obesity. Evidence suggests that existing government policies and industry self-regulatory programs in the United States (US) do not adequately protect young Americans from celebrity endorsement that promotes energy-dense and nutrient-poor food and beverage products. This PhD dissertation describes four studies that examined the nature, extent, and perceived influence of celebrity endorsement used by business firms and non-profit organizations to promote branded food and beverage products or branded campaigns to American children, adolescents and millennial adults to promote healthy food environments. Study one created a database of US celebrities (n=732) involved with food and beverage group, brand or product endorsements (1990-2017) to examine celebrity profiles and their endorsement relationships. Evidence was analyzed using Python data visualization tools and SPSS statistical software. Study two examined celebrities associated with the Partnership for a Healthier America's Fruits and Veggies (FNV) Campaign (2015-2016). Evidence was analyzed using baseline celebrity data (n=82) provided by the campaign initiator, and results informed the FNV Campaign. Study three used Q methodology to explore the views of Millennials, born 1981-1994 (n=40) at Virginia Tech about celebrity endorsement of food and beverage products and brands. Participants sorted 48 celebrity images depicting brand and product endorsements based on celebrity trustworthiness and complete a post Q-sort questionnaire. Data analysis used PQMethod 2.35 statistical software to generate shared viewpoints. Study four identified relevant evidence about US celebrity endorsement of food and beverage products (2000-2019), used an accountability framework to evaluate the adequacy of accountability structures, and suggested future policies and action needed for diverse stakeholders to use celebrity endorsement to promote healthy food environments. Future research should explore the trend of celebrity endorsement used in the US marketplace and examine the influence of this marketing strategy on young people's food beverage preferences and choices based on more representative experimental research.

Estrogen can elicit pleiotropic cellular responses via a diversity of estrogen receptors (ERs)-mediated genomic and rapid non-genomic mechanisms. Unlike the genomic responses, where the classical nuclear ER alpha and ER beta act as transcriptional factors following estrogen binding to regulate gene transcription in estrogen target tissues, the non-genomic cellular responses to estrogen are believed to start at the plasma membrane, leading to rapid activation of second messengers-triggered cytoplasmic signal transduction cascades. The recently acknowledged ER, GPR30 or GPER, was discovered in human breast cancer cells two decades ago and subsequently in many other cells. Since its discovery, it has been claimed that estrogen, ER antagonist fulvestrant, as well as some estrogenic compounds can directly bind to GPER, and therefore initiate the non-genomic cellular responses. Various recently developed genetic tools as well as chemical ligands greatly facilitated research aimed at determining the physiological roles of GPER in different tissues. However, there is still lack of evidence that GPER plays a significant role in mediating endogenous estrogen action in vivo. This review summarizes current knowledge about GPER, including its tissue expression and cellular localization, with emphasis on the research findings elucidating its role in health and disease. Understanding the role of GPER in estrogen signaling will provide opportunities for the development of new therapeutic strategies to strengthen the benefits of estrogen while limiting the potential side effects.

Type 2 Diabetes Mellitus (T2D) is a major public health issue. Diabetes prevalence is growing and is the 7th leading cause of death in the US. Virginia has a slightly higher prevalence than the national average. Community-based diabetes lifestyle management programs that include a physical activity component are effective at improving glycemic control and influencing lifestyle behavior changes among people with T2D. The Balanced Living with Diabetes (BLD) program uses an active learning approach to improve glycemic control and healthful lifestyles. The Lifelong Improvements through Fitness Together (LIFT) program uses behavioral strategies to improve functional fitness, participant engagement, and program adherence. Participant retention is a challenge for community-based program. Participants may start the program, but then fail to complete the program and/or adhere to program recommendations. Two pilot studies were conducted to evaluate strategies for increasing participant retention and improving outcomes. The first evaluated the implementation and impact of a retention plan implemented in BLD programs conducted from 2015-2017. There were more participants returned to the reunion class session prior the development of the retention plan. There were improvements in some health behavior and self-efficacy indicators in programs that used the retention materials, however, impacts on outcomes were mixed. Preliminary findings showed that the retention plan could be a tool for providing additional support to participants, however strategies for dissemination of the retention plan needs to be reevaluated. The second study evaluated the impact on participant engagement and program outcomes when the LIFT program is incorporated with the BLD program. There was an increase in participant retention, self-efficacy and one health behavior for physical activity and health behaviors related to diet in BLD+LIFT programs. Extension Agents are willing to implement the BLD+LIFT programs, however, clarity of program logistics is needed prior to implementation. Extension Agents indicated that implementation of these programs in locations with older adults that have greater disability may not be the best locations. Having larger studies on the effect of incorporation of LIFT with the BLD with older adult populations that have fewer co-morbidities are needed to determine the impact of addition of the LIFT program with the BLD program on program outcomes. Doctor of Philosophy Type 2 Diabetes is a chronic disease in which the body does not use insulin as it should or does not produce enough insulin. The Centers for Disease Control and Prevention (CDC) indicates that the prevalence of diabetes was 23.1 million among US adults in 2015. The prevalence of diabetes in Virginia is slightly higher than the national average. Community-based diabetes lifestyle management programs that include a physical activity component are effective in improving glycemic control and influencing lifestyle behavior changes among people with T2D. The Balanced Living with Diabetes (BLD) program uses an active learning approach to influence better glycemic control and healthful lifestyles. The Lifelong Improvements through Fitness Together (LIFT) program uses behavioral strategies to improve functional fitness, participant engagement, and program adherence. The challenge with implementing community-based programs is retention. Participants may initially agree to participate in a program, but then fail to complete the program and/or follow program recommendations. Two pilot studies were conducted to evaluate strategies for increasing participant retention and program outcomes. The first evaluated the implementation and impact of a retention plan implemented in BLD programs conducted from 2015-2017. There were more participants returned prior to the development of the retention plan. There were improvements in some health behavior and self-efficacy indicators in programs that used the retention materials, however, impacts on outcomes were mixed. Findings showed that the retention plan could be a useful tool for providing additional support to participants, however distribution of the retention plan needs to be reevaluated. The second study evaluated the impact of program outcomes when the LIFT program is combined with the BLD program. There was an increase in participants' self-confidence to perform physical activity and making changes in their diet in BLD+LIFT groups. Extension Agents were enthusiastic about conducting more BLD+LIFT programs, however, program procedures and the time commitment needs to be understood before doing so. Expanding the conduct of BLD programs that incorporate the LIFT program can be effective in improving glycemic control and increasing physical activity, however, working with organizations that service people that have diabetes with fewer health conditions that limit physical activity may be more effective.

Unhealthy food and beverage products marketed through diverse media platforms contribute to poor diet quality, obesity, and chronic diseases in the United States (US). Limited empirical research is available on the effectiveness of integrated marketing communications (IMC) and branded media campaigns used to positively influence Americans' diet-related cognitive, behavioral and health outcomes. The purpose of this research was to assess and inform the Partnership for a Healthier America's Fruits and Veggies (FNV) Campaign to increase fruit and vegetable purchases and intake among targeted teen and mom populations. This PhD research evaluated the use, outcomes, and perspectives on branded food marketing campaigns to inform the FNV Campaign through four studies. Study 1 used a mixed-methods research approach that included a scoping review; comprehensive review of peer-reviewed articles, gray literature and media sources; and key-informant interviews (n=11) to identify 13 national branded marketing campaigns that used health-branding strategies to encourage healthy dietary purchases and consumption in the US between 1990-2016. Study 2 explored views of diverse stakeholders (n=22) regarding the FNV Campaign using semi-structured interviews that were thematically analyzed using a conceptual framework with eight constructs (i.e., campaign design, reach, adoption, effectiveness, impact, expansion, scaling up and sustainability). Study 3 assessed FNV Campaign awareness, cognitions and behaviors for buying and eating fruits and vegetables through a survey of targeted teen and millennial mom audiences (n=1,604) in Hampton Roads, Virginia and Fresno, California where the FNV Campaign pilot was launched. Study 4 encompassed a mixed-methods evaluation using data from the second and third studies that assessed and explored congruence between FNV brand awareness and recall of IMC strategies among survey respondents with stakeholders' perspectives regarding IMC strategy contexts and potential. This body of research expands knowledge about the effectiveness of diet-related IMC and media campaigns and provides a foundation for future FNV Campaign evaluations. Taken together, findings across the four studies identified significant opportunities for large-scale IMC campaigns to improve evaluation designs and outcome reporting. These results can inform evidence-based strategies and policies to improve the design of the FNV Campaign and other IMC initiatives to improve dietary patterns and population health. Doctor of Philosophy Marketing promotions for unhealthy foods and beverages are widespread in the United States (US) and influence Americans' diet choices that lead to weight gain, obesity, and chronic diseases. There is little understanding of how marketing and media campaigns can be used to positively influence Americans' to consume high quality diets and improve their health outcomes. The purpose of this research was to assess and inform the Partnership for a Healthier America's Fruits and Veggies (FNV) Campaign to increase fruit and vegetable purchases and intake among target populations. This PhD research evaluated the use, outcomes, and perspectives on branded food marketing campaigns to inform the FNV Campaign through four studies. Study 1 used a mixed-methods research approach that included a scoping review of the academic and non-academic literature, and key-informant interviews to identify 13 national branded marketing campaigns that used branding strategies to encourage healthy diets in the US between 1990-2016. Study 2 used interviews to explore the views of 22 stakeholders experienced with the FNV Campaign or fruit and vegetable promotion. Interviews were thematically analyzed using a framework with eight constructs (i.e., campaign design, reach, adoption, effectiveness, impact, expansion, scaling up and sustainability) to understand diverse stakeholder perspectives. Study 3 assessed FNV Campaign awareness, cognitions and behaviors for buying and eating fruits and vegetables through a survey of targeted teen and millennial mom audiences (n = 1,604) in Hampton Roads, Virginia and Fresno, California where the was initially launched. Study 4 used data from the second and third studies that assessed similarities and differences between FNV brand awareness and recall of marketing strategies among survey respondents with stakeholders' perspectives regarding marketing strategy contexts and potential. This body of research expands knowledge about the effectiveness of diet-related marketing and media campaigns and provides a foundation for future FNV Campaign evaluations. Taken together, findings across the four studies identified opportunities for large-scale marketing campaigns to improve evaluation designs and outcome reporting. These results can inform future strategies and policies to improve the design of the FNV and other IMC campaigns to improve population diets and health.

Published version

In July 2018, the United States Department of Housing and Urban Development (HUD) implemented a mandatory smoke-free rule in public housing. This study assessed administrator and resident perceptions of rule implementation during its initial year in the District of Columbia Housing Authority (DCHA). Assessment included nine focus groups (n = 69) with residents and in-depth interviews with administrators (n = 7) and residents (n = 26) from 14 DCHA communities (family = 7 and senior/disabled = 7). Semi-structured discussion guides based on the multi-level socio-ecological framework captured dialogue that was recorded, transcribed verbatim, and coded inductively. Emerging major themes for each socio-ecological framework level included: (1) Individual: the rule was supported due to perceived health benefits, with stronger support among non-smokers, (2) Interpersonal: limiting secondhand smoke exposure was perceived as a positive for vulnerable residents, (3) Organizational: communication, signage, and cessation support was perceived as a need, (4) Community: residents perceived mobility, disability, weather, and safety-related issues as barriers, and (5) Public Policy: lease amendments were perceived as enablers of rule implementation but expressed confusion about violations and enforcement. A majority of administrators and residents reported favorable implications of the mandated HUD rule. The novel application of a socio-ecological framework, however, detected implementation nuances that required improvements on multiple levels, including more signage, cessation support, clarification of enforcement roles, and addressing safety concerns.

Background Food waste studies have been used for more than 40 years to assess nutrient intake, dietary quality, menu performance, food acceptability, cost, and effectiveness of nutrition education in the National School Lunch Program (NSLP). Objective Describe methods used to measure food waste and respective results in the NSLP across time. Methods A systematic review using PubMed, Science Direct, Informaworld, and Institute of Scientific Information Web of Knowledge was conducted using the following search terms: waste, school lunch, plate waste, food waste, kitchen, half method, quarter method, weight, and photography. Studies published through June 2015 were included. The systematic review followed preferred reporting items for systematic reviews and meta-analyses recommendations. Results The final review included 53 articles. Food waste methodologies included inperson visual estimation (n = 11), digital photography (n = 11), direct weighing (n = 23), and a combination of in-person visual estimation, digital photography, and/or direct weighing (n = 8). A majority of studies used a preepost intervention or cross-sectional design. Fruits and vegetables were the most researched dietary component on the lunch tray and yielded the greatest amount of waste across studies. Conclusions Food waste is commonly assessed in the NSLP, but the methods are diverse and reporting metrics are variable. Future research should focus on establishing more uniform metrics to measure and report on food waste in the NSLP. Consistent food waste measurement methods will allow for better comparisons between studies. Such measures may facilitate better decision making about NSLP practices, programs, and policies that influence student consumption patterns across settings and interventions. National Institute of General Medical Sciences of the National Institutes of Health [P20GM103474, 5P20GM104417]; Cornell Behavioral Economics in Child Nutrition Programs (BEN) Center Grants Program [77867-10660] Research reported in this publication was partially supported by the National Institute of General Medical Sciences of the National Institutes of Health under award nos. P20GM103474 and 5P20GM104417. Research reported in this publication was partially supported by Cornell Behavioral Economics in Child Nutrition Programs (BEN) Center Grants Program under award no. 77867-10660. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or Cornell BEN Center.

Type 2 diabetes (T2D) is one of the fasting growing chronic diseases, caused by insulin resistance and pancreatic β-cell dysfunction. While over thirty medications were approved to treat T2D in the United States, less than one in four patients treated with anti-diabetic drugs achieved the glycemic target. Thus, identifying more effective anti-diabetic drugs is still needed for improving glycemic control in T2D patients. Incretins are gut hormones that possess potent insulinotropic action, which have drawn considerable attention in research and developing treatment strategy for T2D. Specifically, glucagon like peptide 1 (GLP-1), the most important incretin that is secreted from enteroendocrine L-cells in response to food ingestion, plays a vital role in maintaining glycemic homeostasis via potentiating glucose stimulated insulin secretion (GSIS) and promoting pancreatic β-cell proliferation and survival. Therefore, targeting L-cells to induce GLP-1 secretion would be an alternative strategy for treating T2D. The goal of this research was to identify low-cost and safe naturally occurring agents as a primary or adjuvant treatment for T2D. Here, I found that a small molecule, elenolic acid (EA), which was generated in our lab but is also present in mature olive and extra virgin olive oil, dose-dependently stimulated GLP-1 secretion in mouse clonal L-cells and isolated mouse ileum crypts. EA induced a rapid increase in intracellular [Ca2+]i and the production of inositol trisphosphate in L-cells, indicating that EA activates phospholipase C (PLC)-mediated signaling. Consistently, inhibition of (PLC) ablated EA-stimulated increase of [Ca2+]i and GLP-1 secretion in L-cells. In addition, EA-triggered GLP-1 secretion from L-cells was blocked by YM-254890, a Gαq inhibitor. Consistent with our in vitro study, a single dose of EA acutely stimulated GLP-1 secretion in mice, accompanied with an improved oral glucose tolerance. Chronic administration of EA restored the impaired glucose and lipid homeostasis in DIO mice, which may be partially due to promoting GLP-1 secretion and reduced hepatic gluconeogenesis. In addition, EA suppressed appetite, reduced food intake and gastric emptying rate, as well as promoted weight loss in obese mice, demonstrating that it is also an anti-obesity agent. Further, EA treatment reduced lipid absorption, and promoted hepatic fatty acid oxidation, and reversed abnormal plasma lipid profiles in DIO mice. Consistently, EA exerted potent anti-diabetic action in db/db mice, and its blood glucose-lowering effect is comparable with that of liraglutide in blood glycemic control but is better than that of metformin in this overt diabetic model. Collectively, I have identified for the first time, as to the best of our knowledge, that EA could be a dual-action compound that exerts anti-diabetic effects via activation of the GLP-1 mediated metabolic pathway and suppression of hepatic gluconeogenesis, leading to effective control on food intake, body weight gain, and glycemia in T2D mice. Doctor of Philosophy Type 2 diabetes (T2D) is one of the fasting growing chronic diseases, which results from insulin resistance and pancreatic β-cell dysfunction. Even though there have been over thirty drugs approved to treat T2D in the United States, less than 25% of patients treated with anti-diabetic drugs achieved the glycemic target. Thus, more effective anti-diabetic drugs are still needed for improving glycemic control in patients with T2D. Incretins are a group of gut hormones and responsible for over 50% postprandial insulin secretion in humans, which have drawn considerable attention in research and developing a treatment strategy for T2D. Specifically, glucagon-like peptide 1 (GLP-1), the most important incretin that is secreted from enteroendocrine L-cells in response to food ingestion, plays a vital role in controlling blood glucose via potentiating glucose-stimulated insulin secretion (GSIS) and promoting pancreatic β-cell proliferation and survival. Therefore, targeting L-cells to induce GLP-1 secretion would be an alternative strategy for treating T2D. The goal of this research was to identify low-cost and safe naturally occurring agents as a primary or adjuvant treatment for T2D. Here, I found that a small molecule, elenolic acid (EA), which was synthesized in our lab but is also present in mature olive and extra virgin olive oil, dose-dependently stimulated GLP-1 secretion in mouse clonal L-cells and isolated mouse ileum crypts (containing L-cells). Further experiments showed that EA induced a rapid increase in intracellular [Ca2+]i and the production of inositol trisphosphate (IP3) in L-cells, indicating that EA activates phospholipase C (PLC)-mediated signaling, as IP3 is a direct product of PLC. Consistently, inhibition of PLC ablated EA-stimulated increase of [Ca2+]i and GLP-1 secretion in L-cells. In addition, EA-triggered GLP-1 secretion from L-cells was blocked by YM-254890, a Gαq inhibitor. In line with the in vitro study, a single dose of EA acutely elevated plasma GLP-1 concentration in mice, accompanied by improved oral glucose tolerance. Chronic administration of EA restored the impaired glucose and lipid homeostasis in diet-induced obese (DIO) mice, which may be partially due to promoting GLP-1 secretion and reduced hepatic gluconeogenesis. In addition, EA suppressed appetite, reduced food intake, and gastric emptying rate, as well as promoted weight loss in the DIO mice, demonstrating that it is also an anti-obesity agent. Further, EA treatment reduced lipid absorption and promoted hepatic fatty acid oxidation, as well as reversed abnormal plasma lipid profiles in the DIO mice. Consistently, EA exerted potent anti-diabetic action in predisposed diabetic mice (db/db), and its blood glucose-lowering effect is comparable with that of liraglutide, a commercial GLP-1 receptor agonist, in blood glycemic control but is better than that of metformin, a widely used first-line anti-diabetic drug, in this overt diabetic model. Collectively, I have identified for the first time, as to the best of our knowledge, that EA could be a dual-action compound that exerts anti-diabetic effects via activation of the GLP-1 mediated metabolic pathway and suppression of hepatic gluconeogenesis, leading to effective control on food intake, body weight gain, and glycemia in T2D mice.

Duchenne Muscular Dystrophy (DMD) is an X-linked recessive, progressive muscle-wasting disease characterized by mutations in the dystrophin gene. Duchenne muscular dystrophy is the most common and most severe form of inherited muscle diseases, with an incidence of 1 in 3,500 male births1,2. Mutations in the dystrophin gene result in non-functional dystrophin or the complete absence of the protein dystrophin, resulting in necrosis and fibrosis in the muscle, loss of ambulation, cardiomyopathies, inadequate or failure of respiratory function, and decreased lifespan. Although there has been little research for effective nutritional strategies, dietary intervention may be effective as an adjuvant treatment. In this study, wild type (WT) and mdx animals were provided either a control or elevated branched chain amino acid (BCAA) diet nocturnally for 25 weeks to determine if the elevated BCAAs would attenuate muscle torque loss. Twenty-five weeks of chronic, elevated BCAA supplementation had no impact on muscle function measures. Interestingly, mdx and WT animals had the same torque responses in the low stimulation frequencies (1 Hz – 30 Hz) compared to higher stimulation frequencies. Tetanus was reached at a much lower stimulation frequency in mdx animals compared to WT animals (100 Hz vs +150 Hz). The mdx mouse consistently had more cage activity in the light cycle X- and Y-planes. Interestingly, animals on the BCAA diet increased X-, Y-, and Z-plane activity in the dark cycles at four weeks while animals on the control diet more Z-plane activity at 25 weeks, although not significant. All three BCAAs were elevated in the plasma at 25 weeks, although only Leu was significantly elevated. The BCAAs had no effect on. The diaphragm and skeletal muscle masses were larger in mdx animals, and WT animals had a significantly larger epididymal fat pad. The active state of BCKDC determined by phosphorylation of the E1α enzyme was greater in WT animals in white skeletal muscle, but not red skeletal muscle. Protein synthesis effectors of the mTORC1 signaling pathway and autophagy markers were similar among groups. Wild type animals had increased mTORC1 effectors and animals on the BCAA diet had decreased autophagy markers, although not significant. Although BCAAs did not affect muscle function, fibrosis, or protein synthesis effectors, this study illustrates the functionality of mdx muscles over time. It would be interesting to see how the different muscle fiber types are affected by DMD, noting the differences between the diaphragm, heart, red muscle, and white muscle fibrosis markers. Although there was no increase in mTORC1 effectors with an elevated BCAA diet, it would be interesting to determine muscle protein synthesis, myofibrillar protein synthesis, and total protein turnover in the mdx mouse with an elevated BCAA diet, although the dietary intervention started when mice arrived at 4 weeks of age, earlier intervention may be beneficial early in the disease process. Doctor of Philosophy Duchenne Muscular Dystrophy (DMD) is an X-linked recessive, progressive muscle-wasting disease characterized by mutations in the dystrophin gene. Duchenne muscular dystrophy is the most common and most severe form of inherited muscle diseases, with an incidence of 1 in 3,500 male births1,2. Mutations in the dystrophin gene result in non-functional dystrophin or the complete absence of the protein dystrophin, resulting in necrosis and fibrosis in the muscle, loss of movement and walking ability, cardiomyopathies, inadequate or failure of respiratory function, and decreased lifespan. Although there has been little research for effective nutritional strategies, dietary intervention may be effective as an adjuvant treatment and palliative care. The branched chain amino acids (BCAAs) are known to directly stimulate muscle protein synthesis by direct activation of the mechanistic target of rapamycin complex 1 (mTORC1). This study aimed to illustrate the differences between diseased and healthy mice and determine if BCAAs can reduce muscle torque loss. Twenty-five weeks of chronic, elevated BCAA supplementation had no impact on muscle function measures. Interestingly, mdx and WT animals had the same torque responses in the low stimulation frequencies (1 Hz – 30 Hz) compared to higher stimulation frequencies. Tetanus was reached at a much lower stimulation frequency in mdx animals compared to WT animals (100 Hz vs +150 Hz). The mdx mouse consistently had more cage activity in the light cycle X- and Y-planes. Interestingly, animals on the BCAA diet increased X-, Y-, and Z-plane activity in the dark cycles at four weeks while animals on the control diet more Z-plane activity at 25 weeks, although not significant. All three BCAAs were elevated in the plasma at 25 weeks, although only Leu was significantly elevated. The BCAAs had no effect on. The diaphragm and skeletal muscle masses were larger in mdx animals, and WT animals had a significantly larger epididymal fat pad. The active state of BCKDC determined by phosphorylation of the E1α enzyme was greater in WT animals in white skeletal muscle, but not red skeletal muscle. Protein synthesis effectors of the mTORC1 signaling pathway and autophagy markers were similar among groups. Wild type animals had increased mTORC1 effectors and animals on the BCAA diet had decreased autophagy markers, although not significant. Although BCAAs did not affect muscle function, fibrosis, or protein synthesis effectors, this study illustrates the functionality of mdx muscles over time. It would be interesting to see how the different muscle fiber types are affected by DMD, noting the differences between the diaphragm, heart, red muscle, and white muscle fibrosis markers. Although there was no increase in mTORC1 effectors with an elevated BCAA diet, it would be interesting to determine muscle protein synthesis, myofibrillar protein synthesis, and total protein turnover in the mdx mouse with an elevated BCAA diet, although the dietary intervention started when mice arrived at 4 weeks of age, earlier intervention may be beneficial early in the disease process.

Type 2 Diabetes Mellitus (T2DM), a metabolic disorder denoted by elevated blood glucose levels and insufficient insulin action, is growing in prevalence worldwide . Barriers to improving disease outcome resolve primarily around identifying and intervening during the preliminary stages of insulin resistance, a state clinically referred to as pre-diabetes. Emerging evidence suggests that mitochondrial dysfunction may underlie , and potentially precede, progressive insulin resistance, suggesting that biomarkers indicative of mitochondrial dysfunction could predict disease risk and status. In this study, we examined epigenetic modifications, in the form of DNA methylation, in the promoter region of peroxisome proliferator activated receptor gamma coactivator 1 alpha (PGC-1α), a known regulator of mitochondrial biogenesis. Following the initiation of a high fat diet, we observed significant genotypic (DNA methylation) and phenotypic (mitochondrial copy number) alterations in C57/BL6 rodent models. These changes preceded overt disease onset, as classified by clinically utilized indices, which included the homeostatic model assessment for insulin resistance (HOMA-IR), the homeostatic model assessment for β-cell dysfunction (HOMA- β), and the quantitative insulin-sensitivity check index (QUICKI). Our data indicate that methylation analysis may serve as an effective clinical parameter to use in conjunction with physiological criterion for the diagnosis of pre-diabetes and the assessment of T2DM disease risk, and adds to the growing body of work seeking to elucidate the role. Doctor of Philosophy High blood glucose, referred to as type 2 diabetes (T2DM), increases the risk for heart and kidney disease, blindness, stroke, and death. Efforts to prevent T2DM have centered primarily around behavioral interventions, which include increased physical activity and decreased caloric intake. Importantly, the interventions are most effective when implemented early on in disease progression. In this study, we sought to examine the effects of a high fat diet on the epigenetic profile of PGC-1α, a gene responsible for maintaining mitochondrial biogenesis. The mitochondria, the powerhouse of the cell, is responsible for maintaining the energy systems in the body. Therefore, we examined how increasing in caloric intake resulted in changes in the epigenetic profile of the PGC-1α promoter, and how these changes impacted mitochondrial number. Further, we sought to examine how hypermethylation of PGC-1α led to changes in gene and protein expression in the mitochondria. Results from our study indicate that DNA methylation changes preceded disease onset, as characterized by the homeostatic model assessment for insulin resistance (HOMA-IR), the homeostatic model assessment for β-cell dysfunction (HOMA- β), and the quantitative insulin-sensitivity check index (QUICKI). Our data indicate that methylation analysis may serve as diagnostic and risk assessment tool for pre-diabetes and T2DM in conjunction with physiological measures.

With 47% of the veteran population aged 65 years or older, an age group with the highest prevalence of type 2 diabetes (T2D), prevention of T2D is critical within the Veterans Health Administration (VHA) system (1). Resist Diabetes (RD), a Social Cognitive Theory-based resistance training program, has been shown to reduce the prevalence of prediabetes by 34% in prediabetic older adults (2). The RD program could fill the gap within the VHA system for a structured diabetes prevention program not focused on weight, but instead focused on exercise. This thesis describes a mixed-methods investigation to determine what adaptations to the RD program are needed to implement the program within the Salem Veterans Affairs Medical Center (VAMC) patients. Salem VAMC care providers and veteran patients were recruited to gauge perceptions of the RD program, and identify barriers and facilitators to the referral process and program implementation. Semi-structured open-ended questionnaires and interview guides based upon the Consolidated Framework for Implementation Research (CFIR) (3) were utilized to determine major and minor themes within the provider and veteran responses. Quantitative data was also obtained to assess provider perceptions and participant characteristics. Major findings included: positive perceptions by providers for RD program acceptability, appropriateness and feasibility (4) within the Salem VAMC, positive perceptions of Intervention Characteristics of RD by veterans, major barriers to participation lied within the Outer Setting domain with travel, transportation, and time constraints. Major facilitators to program implementation lied within the Inner Setting domain with promotion by primary care. These findings could be used to inform a future version of the RD program adapted for the VHA system. Master of Science With 47% of the veteran population aged 65 years or older, an age group with the highest prevalence of type 2 diabetes (T2D), prevention of T2D is critical within the Veterans Health Administration (VHA) system. Resist Diabetes, a Social Cognitive Theory-based resistance training program, reduced the prevalence of prediabetes by 34% in prediabetic older adults in the Roanoke, VA area. This thesis describes a mixed-method investigation to determine what adaptations to the Resist Diabetes program are needed to implement the program within the Salem Virginia Veterans Affairs Medical Center.

Childhood (ages 2-9 years) and adolescence (ages 10-19 years) make up important periods of human life. Food preferences and dietary behaviors are developing during this time, and health status can impact noncommunicable disease (NCD) risk in adulthood. Many of the behaviors associated with developing NCD are modifiable and include dietary behaviors. Globally, children and adolescents are impacted by NCD, and obesity prevalence is growing among this population. One of the recommendations for combating obesity is to promote the consumption of healthy foods and beverages and the reduction of sugar-sweetened beverage intake. In order to assess beverage patterns, the Healthy Beverage Index (HBI) was created for adults. This tool provides a score from 0-100, with higher scores indicating better adherence to evidence-based beverage recommendations. Because many beverage recommendations for children and adolescents differ from those for adults, the purpose of this thesis was to develop the HBI for U.S. Children and Adolescents (HBI-CA). National Health and Nutrition Examination Survey (NHANES) data was used to calculate HBI-CA scores among a sample of U.S. children and adolescents. The HBI-CA resulted in scores similar to the adult HBI and provides a reference of beverage pattern quality in this population. Significant differences in scores were found among males and females and certain races. Evaluation of the HBI-CA demonstrated its construct validity and reliability. The HBI-CA still needs to undergo predictive validity tests. Overall, the HBI-CA provides a method to assess beverage pattern quality in clinical, public health, and research settings for this important population. Master of Science Children (ages 2-9 years) and adolescents (ages 10-19 years) makeup an important population for the implementation of dietary interventions because they are still developing their food preferences and dietary behaviors. The rise of obesity among these age groups indicates that their dietary behaviors may not be aligned with recommendations. Beverages can easily be overlooked as a dietary component, but they currently constitute a large portion of daily energy intake. In order to assess beverage pattern quality in this population, the Healthy Beverage Index for U.S. Children and Adolescents (HBI-CA) was developed based on the HBI for adults. This tool provides a score from 0-100, with higher scores indicating better adherence to evidence-based beverage recommendations. The HBI-CA also provides a resource of comprehensive beverage recommendations for these age groups, which was previously limited in the literature. National Health and Nutrition Examination Survey (NHANES) data was used to calculate HBICA scores among a sample of U.S. children and adolescents. The HBI-CA resulted in scores similar to the adult HBI and provides a reference of beverage pattern quality in this population. Significant differences in scores were found among males and females and certain races. Evaluation of the HBI-CA demonstrated its ability to reliably and consistently measure beverage pattern quality. Future evaluation of the HBI-CA should include examination of the relationship between HBI-CA scores and dietary-related health outcomes. Overall, the HBI-CA provides a method to assess beverage pattern quality in clinical, public health, and research settings for this important population.

The accurate measurement of total energy expenditure is a cornerstone of metabolic research. However, there is a lack of measurement methods that are valid, objective, inexpensive, and easy to use. Accelerometry, along with validated prediction equations for resting energy requirements, may provide an opportunity to fill this void. Twenty weight stable adults (12 female, 8 male) who recently participated in a controlled feeding study comprised the study sample. Total energy requirements were assessed from the controlled feeding period in which weight stability was achieved using the intake-balance method. Resting energy expenditure was assessed using the Mifflin-St. Jeor equation. Participants wore accelerometers to objectively assess habitual physical activity. The accelerometer data obtained along with subjects' demographic and biometric data were used to predict non-resting energy expenditure (NREE) using step-wise linear regression in JMP. Bland-Altman plots and Spearman's Rho correlations were used to determine the validity of the total energy requirements obtained from the sum of the predicted non-resting energy expenditure. Estimated resting energy expenditure was compared with the total energy requirements assessed using the intake-balance method from the controlled feeding period. The resulting prediction equation is as follows: 480.93 – 180.69(sex) + 0.21(Accelerometer kcals) + 617.98(BF%) = AEE. The sex was coded as 1 for females and 0 for males. This prediction model has a coefficient of determination of 0.74 (0.70 adjusted). On average, the model overestimates AEE by 76 kcals. This new model could be the key to accurately, inexpensively and objectively measuring total energy requirements. Master of Science Accurate measurement of the total amount of energy (i.e. calories) utilized by the body throughout the day, also known as total energy expenditure, is a vital component of metabolic research. However, there is a lack of measurement methods that are valid, objective, inexpensive, and easy to use. Accelerometers combined with equations designed to predict total energy expenditure may be able to fill this gap. Accelerometers are devices worn on the body that measure accelerative forces from physical activity. Twenty weight stable adults (12 female, 8 male), who recently participated in a study in which all dietary intake and exercise were closely monitored (controlled feeding study), comprised the study sample. The amount of energy needed to maintain weight (total energy requirements) was assessed from the controlled feeding period in which weight stability was achieved. Resting energy expenditure, the energy burned while the body is at rest, was assessed using an equation often used to estimate energy expenditure, the Mifflin-St. Jeor equation. Participants wore accelerometers to objectively assess habitual physical activity. The accelerometer data obtained along with subjects’ demographic (age, sex) and biometric (height, weight, BMI, etc.) data were used to predict non-resting energy expenditure (resting energy expenditure subtracted from total energy expenditure). Multiple statistical tests were used to determine the validity of the total energy requirements obtained from the sum of the predicted non-resting energy expenditure (NREE) and resting energy expenditure. Estimated resting energy expenditure was compared with the total energy requirements assessed using the intake-balance method from the controlled feeding period. The resulting prediction equation is as follows: 480.93 – 180.69(sex) + 0.21(Accelerometer kcals) + 617.98(BF%) = NREE. The sex was coded as 1 for females and 0 for males. This prediction model has a coefficient of determination of 0.74 (0.70 adjusted), which means 70% of the variation in non-resting energy expenditure was explained by changes in the variables in the equation. On average, the model overestimates NREE by 76 Calories per day. This new model could be the key to accurately, inexpensively and objectively measuring total energy requirements.

Recent studies showed that GPR30, a seven-transmembrane G protein-coupled receptor, is a novel estrogen receptor (ER) that mediates some biological events elicited by estrogen in several types of cancer cells. However, its physiological or pathological role in vivo is unclear. For the first project of my dissertation, I investigated the physiological role(s) of GPR30 in energy metabolism by using transgenic mouse model as well as immortalized cell lines and primary stromal cells. We discovered for the first time that GPR30 knockout (GPRKO) female mice were protected from high-fat diet (HFD)-induced obesity, glucose intolerance, and insulin resistance. The decreased body weight gain in GPRKO female mice is due to the reduction in body fat mass. These effects occurred in the absence of significant changes in food intake, intestinal fat absorption, or triglyceride metabolism. However, GPR30 had no significant metabolic effects in male mice fed the HFD and both sexes of mice fed a chow diet. Further, GPR30 expression levels in fat tissues of WT obese female mice greatly increased, whereas ERα/β expression was not altered. Deletion of GPR30 reduced adipogenic differentiation of adipose tissue-derived stromal cells. Conversely, activation of GPR30 enhanced adipogenic differentiation of 3T3-L1 preadipocytes. For the second project, I explored whether estrogen acts through GPR30 to affect adiposity in female mice. For this study, I generated and examined three independent transgenic mouse models, aromatase (Ar) knockout (ArKO) mice, GPRKO, and GPR30 and Ar double knockout (DKO) mice. We discovered that GPR30 deficiency had limited effects on energy metabolism in mice fed a standard chow diet (STD). However, deletion of GPR30 promoted metabolic flexibility in both genders fed a HFD regardless of the presence of estrogen, suggesting that GPR30 may not solely act as an ER. Consistent with our previous findings, GPRKO mice had higher body temperature, indicating that GPR30 deficiency may promote thermogenesis and energy metabolism, resulting in the reduced fat depots and enhanced metabolic flexibility. For the third project, I further explored whether GPR30 is involved in regulating browning of adipose tissue and thermogenesis in mice. The results show that the expression of UCP-1, the key regulator of thermogenic browning, was higher in the adipose tissue of HFD-fed GPRKO female mice as compared with that of WT mice. Consistently, deletion of GPR30 enhanced mitochondrial respiration in brown adipose tissue (BAT), suggesting that GPR30 deficiency at least partially suppressed the fat accumulation by promoting thermogenesis and dissipating energy. Ex vivo, the expression of thermogenic genes and UCP-1 protein level were upregulated in beige adipocytes differentiated from GPR30-deficient stromal vascular fraction (SVF) cells. These findings provide evidence for the first time that deletion of GPR30 reduces adiposity, promotes white adipose beigeing and thermogenesis, therefore preventing the development of obesity in female mice exposed to excess energy. Further investigations elucidating the underlying mechanism by which GPR30 promotes obesity in females could provide a novel therapeutic target to fight obesity in females. Doctor of Philosophy Estrogen can elicit pleiotropic genomic and rapid nongenomic cellular responses via a diversity of estrogen receptors (ERs). Unlike the genomic responses, where the classical nuclear ERα and ERβ initiate gene transcription in estrogen target tissues, the nongenomic cellular responses to estrogen are believed to start at the plasma membrane, leading to rapid activation of second messengers-triggered cytoplasmic signal transduction cascades. The recently acknowledged ER, GPR30, was discovered in human breast cancer cells two decades ago and subsequently in many other cells. Since its discovery, it has been claimed that estrogen, ER antagonist fulvestrant, as well as some estrogenic compounds can directly bind to GPR30, and therefore initiate the rapid nongenomic cellular responses. We are interested to investigate the physiological role(s) of GPR30 in energy metabolism by using transgenic mouse model as well as immortalized cell lines and primary stromal cells. We discovered for the first time that deletion of GPR30 protects female mice from high fat-diet (HFD)-induced obesity and the expression of GPR30 increased in fat tissues of wild type (WT) obese mice, while no alterations of classical ERα/β observed. Consistently, activation of GPR30 by the selective agonist G-1 promotes adipogenic differentiation of 3T3-L1 preadipocytes. ERα is known to exert a protective effect against excess fat accumulation whereas GPR30 may acts as an “obesity gene” and counteracts the classical ERα’s action in regulating fat metabolism. We speculated that there might be a “Yin-Yang” relationship between GPR30 and ERα regarding their actions in the development of obesity. Therefore, we generated three independent transgenic mouse models, GPR30 and aromatase (Ar) double knockout (DKO), GPR30 knockout (GPRKO), and Ar knockout (ArKO) to test our hypothesis that the excess fat accumulation in HFD-fed WT mice could be, or at least partially, caused by the enhanced estrogen-GPR30 signaling. Ar is the key enzyme that catalyzes the biosynthesis of C18 estrogens from C19 androgens in men and postmenopausal women, thereby the ArKO and DKO mouse models allowed us to investigate the role of GPR30 in the absence of endogenous estrogen. We discovered that GPR30 deficiency had limited effects on energy metabolism in young mice fed a standard chow diet (STD). However, deletion of GPR30 promoted metabolic flexibility in both genders fed a HFD regardless of the presence of estrogen, suggesting GPR30 may not solely acts via the ligation of estrogen. Interestingly, consistent with our previous findings, GPRKO mice had higher body temperature, indicating that GPR30 deficiency may promote thermogenesis and energy metabolism, resulting in the reduced fat depots and enhanced metabolic flexibility. Hence, we explored that deletion of GPR30 exerted thermo-promoting effect via upregulation of the mitochondrial uncoupling protein-1 (UCP-1) and enhanced mitochondrial respiration in brown adipose tissue (BAT). Further, the expression of thermogenic genes were significantly higher in the stromal cells-differentiated beige adipocytes, suggesting that GPR30 deficiency suppressed fat accumulation by promoting thremogenic browning of white adipose tissue (WAT) and dissipating excess energy as heat. In summary, my dissertation work provide valuable insight regarding the role of GPR30 in energy metabolism. Further investigations testing whether GPR30 acts as a pro-obesity gene would facilitate our understanding of obesity development and provide a novel therapeutic target to fight obesity.

Skeletal muscle is a primary contributor to body mass and whole-body energy metabolism. It is an adaptive tissue with the ability to fluctuate in size and mechanical properties in response to stimulus. Health conditions involving chronic elevated inflammation levels often feature metabolic inflexibility and losses in skeletal muscle mass. Mononuclear stem cells, termed satellite cells, are mitotic and serve to donate nuclei to muscle fibers to enable skeletal muscle adaptation. Despite the well-characterized nature of satellite cell activation, proliferation, and differentiation, the underlying mechanisms regulating this process is not fully understood. Recent characterization of cytokines secreted by skeletal muscle in an endocrine type fashion has led to discoveries of inflammatory cytokines influencing satellite cell function. However, how the autocrine production and secretion of these cytokines during proliferation and differentiation in humans and their correlation with myogenic transcription factors is not well understood. Our study used satellite cells cultured from the vastus lateralis of 12 male human research subjects, and ELISA analysis to measure levels of TNF-α and IL-6 across proliferation, early differentiation, and late differentiation. Additionally, mRNA levels of Pax7, MyoD, myogenin, IL-6, TNF-α, and TGF-β were assessed in satellite cells cultured from a subset of two endurance trained and two sedentary individuals from the larger group of 12 human subjects. The novelty of our study is the large number of human research subjects and simultaneous analysis of inflammatory cytokine secretion, mRNA inflammatory cytokine expression, and myogenic transcription factor mRNA expression. Results showed an 83% decrease in IL-6 protein secretion 24 hours after exposure to differentiation media (p-value

Research suggests school-based interventions and health education programs can increase children's fruit and vegetable exposure and consumption to improve their diet quality and reduce the risk of diet-related chronic diseases during adulthood. Nevertheless, children have limited availability and access to fruits and vegetables in school environments, which is a barrier to healthy eating. Farm to school (FTS) programs are one of many synergistic interventions to increase children's intake of locally and regionally produced fruits and vegetables at school. In 2015, the United States (U.S.) Department of Agriculture reported that 42% of U.S. school districts and 57% of Virginia schools participated in FTS programs. However, there is a lack of research on FTS programs in Montgomery County, Virginia. This MS thesis describes a mixed-methods, Q Methodology study to explore the views of diverse stakeholders (n=14) regarding the benefits, opportunities, and challenges to implement FTS programs for children, aged 5-11 years, in primary schools in Montgomery County, Virginia. Objective 1 was used to identify and mapped stakeholders invested in or affected by FTS programs in Virginia. Objective 2 used Q Methodology and factor analysis to identify the views of stakeholders regarding the benefits, opportunities, and challenges of FTS programs in Montgomery County. Results found two factors that summarized all 14 participants' viewpoints. Factor one (n=12), Appreciators of Child-Centered Benefits and factor 2 (n=2), Advocates for Legislative Change. Objective 3 described policies and future actions needed to institutionalize and sustain FTS programs in Montgomery County and southwestern Virginia. Master of Science Research suggests school-based interventions and health education programs can increase children’s fruit and vegetable exposure and consumption to improve their diet quality and reduce the risk of diet-related chronic diseases during adulthood. Nevertheless, children have limited availability and access to fruits and vegetables in school environments, which is a barrier to healthy eating. Farm to school (FTS) programs are one of many synergistic interventions to increase children’s intake of locally and regionally produced fruits and vegetables at school. In 2015, the United States (U.S.) Department of Agriculture reported that 42% of U.S. school districts and 57% of Virginia schools participated in FTS programs. However, there is a lack of research on FTS programs in Montgomery County, Virginia. This MS thesis describes a mixed-methods, Q Methodology study to explore the views of diverse stakeholders (n=14) regarding the benefits, opportunities, and challenges to implement FTS programs for children, aged 5- 11 years, in primary schools in Montgomery County, Virginia. Objective 1 identified and mapped stakeholders invested in or affected by FTS programs in Virginia. Objective 2 used Q Methodology and factor analysis to identify the views of stakeholders regarding the benefits, opportunities, and challenges of FTS programs in Montgomery County. Results found two factors that summarized all 14 participants’ viewpoints. Factor one (n=12), Appreciators of Child-Centered Benefits and factor 2 (n=2), Advocates for Legislative Change. Objective 3 described policies and future actions needed to institutionalize and sustain FTS programs in Montgomery County and southwestern Virginia.

Supplemental Nutrition Assistance Program (SNAP)-authorized store environments could be improved to favor consumer purchase of healthy products. Engaging with the key intermediaries who can use marketing-mix and choice-architecture (MMCA) strategies to encourage low-income consumers to purchase healthy products aligned with the Dietary Guidelines for Americans (DGA), 2015-2020 is essential. This PhD research describes five investigations that explored the perspectives of food store owners, managers, and corporate or independent businesses (e.g., retailers) to inform healthy food retail approaches: (1) a systematic review of the literature (1980-2017) identified social-ecological influencers of food store retailers' decision-making and ability to use MMCA strategies to encourage healthy dietary purchases in the United States (US); (2) SNAP-authorized retailers' perceived feasibility and costs to implement healthy MMCA strategies in rural stores were assessed (n=29); (3) SNAP-authorized retailers' healthy food and beverage perceptions and DGA-aligned product offerings were documented; (4) prevalent SNAP-authorized food store retailers in the US and between two states were identified to inform settings where healthy food retail approaches could reach numerous SNAP consumers; (5) and the availability of corporate social responsibility commitments to use MMCA strategies to improve consumers' diet quality among prevalent SNAP-authorized food store chains was explored. The collective findings from the review and four studies were that multiple social-ecological factors (e.g., skills/knowledge, consumers, suppliers) influenced US retailers' decision-making and ability to use MMCA strategies that favor healthy products. Rural retailers perceived prompting and proximity (e.g., labeling and location) strategies as feasible and less costly compared to other MMCA strategies. Some misalignments of healthy food perceptions and food store availability were identified and indicated a need for trainings to enhance the success of healthy food retail programs. To reach numerous SNAP consumers, healthy food retail programs should target nontraditional (e.g., non-grocery) food stores with varied approaches by state. However, few prevalent SNAP-authorized retailers have made public, voluntary commitments to reduce obesity and may reflect a low readiness to engage in partnerships to establish healthy food retail environments. Future research should document approach to and the impact of using MMCA strategies to encourage healthier consumer purchases on business outcomes among diverse store contexts. Doctor of Philosophy Supplemental Nutrition Assistance Program (SNAP) consumers are low-income Americans who could benefit from exposure to store cues or marketing-mix and choice-architecture (MMCA) strategies that ‘make the healthy choice the easy choice’. However, food store owners, managers, and businesses (e.g. retailers) ultimately decide if they will initiate and continue MMCA changes. This research prioritized the retailer perspective to inform healthy food retail approaches. A systematic review of literature (1980-2017) explored multifaceted influencers of food store retailers’ decision-making and ability or willingness to use MMCA strategies to encourage healthier dietary purchases in the United States (US). Additionally, four original research investigations were conducted: (1) a mixed-methods and cross-sectional investigation in rural, central Virginia assessed SNAP-authorized retailers’ perceived feasibility and costs to implement MMCA strategies that encourage healthy consumer purchases in stores; (2) an investigation of SNAP-authorized retailers’ healthy food and beverage perceptions and healthy food store offerings informed healthy food retail approaches in this setting; (3) an examination of prevalent SNAP-authorized food store retailers in the US and between two US states informed targeted healthy food retail approaches that could benefit numerous SNAP consumers; and (4) the availability of corporate social responsibility commitments to use MMCA strategies to promote consumer health among prevalent SNAP-authorized food store chains was explored. Main findings of these investigations indicated that there are multiple factors (e.g., skills/knowledge, consumers, suppliers) that influence US retailers’ decision-making. Also, rural retailers perceived prompting and proximity (e.g., healthy food labeling and moving healthy foods to an ideal location) strategies as feasible and low cost. There were some misalignments of healthy food perceptions and food store availability, indicating a need for trainings to enhance the likelihood for healthy food retail programs to succeed. In approaching SNAP-authorized retailers for partnership opportunities, targeting nontraditional (e.g., non-grocery) settings may reach more consumers and varied approaches by state are warranted. However, many of these prevalent SNAP retailers lack publicly available commitments to reduce obesity which may hinder partnership approaches. Future work should document approach to and the impact of using MMCA strategies to encourage healthier consumer purchases on diet quality and business outcomes among diverse store contexts.

Around 46.2 million Americans living in rural areas are disproportionately burdened by health disparities. Likewise, obesity and obesity-associated diseases (e.g., diabetes, cardiovascular disease) are much higher for rural residents when compared to their urban counterparts. There is a high need to understand and address the nutritional determinants of these health inequities among adults and adolescents. One area of concern in rural dietary habits pertains to added sugars and more specifically, sugar-sweetened beverages (SSB). Excessive added sugars and SSB intake have been strongly linked to many of the nutrition and chronic disease disparities impacting rural residents. Moreover, studies conducted in rural populations have found high consumptions of these in both adults and adolescents. There is an opportunity to better understand added sugars and SSB patterns in rural populations to inform the development of culturally relevant, multi-level interventions that address high consumption. Study #1 is a cross-sectional study that explores top food and beverage sources of added sugars in the diet of adults (n = 301) living in rural areas of Southwest Virginia. Study #2 uses a nationally representative sample of adolescents (n = 1,560) from the Family Life, Activity, Sun, Health and Eating (FLASHE) study sponsored by the National Cancer Institute, to explore factors across the levels of the socioecological model associated with adolescent SSB intake. Study #3 utilizes focus groups and a pilot trial to understand language preferences, acceptability and use of SMS aimed at caregivers to reduce SSB intake in both caregivers and adolescents living in rural areas of Southwest Virginia (n = 33). Collectively, these three studies offer recommendations and culturally relevant strategies for future large-scale trials aimed at reducing SSB intake among adolescents and caregivers in rural communities and ultimately reducing rural health disparities. Doctor of Philosophy Rural populations in the United States are at higher risk for being diagnosed with and dying from preventable and obesity-associated diseases like heart disease and cancer. Excessive added sugars and sugary drink (i.e. sodas, sweet tea/coffee, energy drinks, sweetened fruit drinks, sports drinks) intake have been strongly linked to many of the chronic diseases afflicting rural residents. Moreover, studies conducted in rural populations have found high consumptions of these, in both adults and adolescents. There is a great need to better understand added sugars and sugary drink patterns in rural populations so that we can develop programs to reduce consumption that are also culturally well received. Study #1 in this dissertation explores top food and beverage sources of added sugars in the diet of 301 adults living in rural areas of Southwest Virginia. Study #2 uses a nationally representative sample of 1,560 adolescents to explain why adolescent SSB intake might be higher. Study #3 aims to understand language preferences, acceptability and use of a text message program to reduce sugary drink intake in both caregivers and adolescents living in rural areas of Southwest Virginia. Collectively, these three studies offer recommendations and culturally relevant strategies for future large scale trials aimed at reducing sugary drink intake among adolescents and caregivers in rural communities and ultimately improving rural health.

Dysfunction within complex I (CI) of the mitochondrial electron transport system has been implicated in a number of disease states ranging from cardiovascular diseases to neuro-ophthalmic indications. Herein, we provide three novel approaches to model and study the impacts of injury on the function of CI. Cardiovascular ischemia/reperfusion (I/R) injury has long been recognized as a leading contributor to CI dysfunction. Aside from the physical injury that occurs in the tissue during the ischemic period, the production of high levels of reactive oxygen species (ROS) upon reperfusion, led by reverse electron transport (RET) from CI, causes significant damage to the cell. With over 700,000 people in the US set to experience an ischemic cardiac event annually, the need for a pharmacological intervention is paramount. Unfortunately, current pharmacological approaches to treat I/R related injury are limited and the ones that have shown efficacy have often done so with mixed results. Among the current approaches to treat I/R injury antioxidants have shown some promise to help preserve mitochondrial function and assuage tissue death. The studies described herein have provided new, more physiologically matched, methods for assessing the impact of potential therapeutic interventions in I/R injury. With these methods we evaluated the efficacy of the coenzyme-Q derivative idebenone, a proposed antioxidant. Surprisingly, in both chemically induced models of I/R and I/R in the intact heart, we see no antioxidant-based mechanism for rescue. The mechanistic insight we gained from these models of I/R injury directed us to further examine CI dysfunction in greater detail. Through the use of two cutting edge genetic engineering approaches, CRISPR/Cas9 and Artificial Site-specific RNA Endonucleases (ASRE), we have been able to directly edit the mitochondria to accurately model CI dysfunction in disease. The use of these genetic engineering technologies have provided first in class methods for modeling three unique mitochondrial diseases. The culmination of these projects has provided tremendous insight into the role of CI in disease and have taken a significant step towards elucidating potential therapeutic avenues for targeting decrements in mitochondrial function. Doctor of Philosophy Within the mitochondria, “the powerhouse of the cell,” exists a series of five enzyme complexes that produce 90% of the energy for our cells need to function. The largest of these enzymes, complex I (CI), plays an important role in ensuring proper mitochondrial function. Injury to CI contributes to a number of diseases, but surprisingly few options exist to treat complex I. One of the most prevalent forms of CI dysfunction can be seen in ischemia/ reperfusion injury, a form of which is most commonly recognized as a heart attack. Surprisingly, the American Heart Association reports that in the next year over 700,000 people in the US will suffer from an ischemic event. With such a profound impact on the population, the need for new therapeutic developments is extremely high. Some current therapeutic approaches have been shown to be effective at treating cardiac dysfunction, but few address the dysfunction that occurs in the mitochondria. Here we test both a method for modeling these ischemia/reperfusion-based injuries and a potential therapeutic for treating these injuries within the context of CI dysfunction. We further evaluate CI dysfunction by using both established genetic engineering approaches as well as a completely new method to model CI disease. Through the use of two cutting edge genetic engineering approaches, we have been able to directly edit components of the mitochondria to accurately model CI dysfunction in disease. The use of these genetic engineering technologies have provided a first-in-class method for modeling three unique mitochondrial diseases. The culmination of these projects has provided tremendous insight into the role of CI in disease and have taken a significant step towards elucidating potential therapeutic avenues for targeting decrements in mitochondrial function.

Mitochondrial structure and function are inextricably linked ("structure-function"), with decrements in structure-function evident across diseases. Barriers to new therapies include a complete understanding of the underlying molecular culprits, as well as effective mitochondria-targeted therapies that mitigate injury. In these works, we investigate the role of cristae-shaping factors like cardiolipin in health and disease. In a series of studies, we tested the effects of the cell-permeable tetrapeptides, elamipretide and a postulated peptide, (arginine-tyrosine-lysine-phenylalanine; "RYKF"), on the recovery of mitochondrial structure-function after injury. Elamipretide is a clinical-stage compound currently under investigation for genetic and age-related mitochondrial diseases, yet the mechanism of action is not completely understood. We used a combination of physiological models, mitochondrial imaging, and biomimetic membrane studies to test the hypothesis that elamipretide and RYKF-cardiolipin interactions improved mitochondrial structure-function. Post-ischemic treatment with elamipretide sustained mitochondrial function across electron transport chain complexes. Endogenous RYKF expression similarly improved mitochondrial respiration after peroxide and hypoxia nutrient deprivation injuries. Using two parallel electron microscopy paradigms, we show elamipretide and RYKF treatment led to maintenance of mitochondrial ultrastructure and notably, improved cristae interconnectedness. Finally, we utilized a novel biomimetic membrane system to model the pathological mitochondrial membrane and found that elamipretide and RYKF both improved biophysical pressure-area relationships through a mechanism that appears to involve aggregating cardiolipin. Our data indicate that targeting pathophysiological mitochondrial membranes with cationic, lipophilic peptides can improve bioenergetics by sustaining cristae networks and support interdependent relationships between mitochondrial structure and function. Doctor of Philosophy Mitochondria, the powerhouses of the cell, form energy networks that produce over 90% of the body’s energy. Mitochondrial dysfunction is implicated across diseases, yet no FDA-approved treatments exist that improve mitochondrial energy production. In this study, we tested the effects of elamipretide, a peptide that localizes to mitochondria. Although elamipretide is currently in clinical trials for several diseases characterized by energetic deficiencies, its mechanism of action is not fully understood. Since mitochondrial structure and function are directly linked, we modeled heart attacks in cultured cells and rat hearts to test the hypothesis that elamipretide and a postulated analog, RYKF, glue damaged mitochondrial membranes back together to preserve structure and function during disease. In hearts subjected to a heart attack, elamipretide significantly protected mitochondrial energy production. Similarly, RYKF protected mitochondrial function in muscle cells exposed to peroxide stress. In damaged hearts imaged with electron microscopy, elamipretide and RYKF treatment significantly improved mitochondrial structure and notably, improved the interconnectedness of mitochondrial energy networks. Furthermore, elamipretide and RYKF improved the integrity of diseased mitochondrial membranes. Together, these data support our hypothesis that elamipretide and RYKF act as mitochondrial adhesion molecules to protect mitochondrial structure and sustain energy production during disease.

Accelerometer data can be analyzed using a variety of methods which are effective in the clinical setting. Time-series analysis is used to analyze spatiotemporal variables in various populations. More recently, investigators have focused on gait complexity and the structure of spatiotemporal variations during walking and running. This study evaluated the use of time-series analyses to determine gait parameters during running. Subjects were college-age female soccer players. Accelerometer data were collected using GPS-embedded trunk-mounted accelerometers. Customized Matlab® programs were developed that included Gaussian continuous wavelet transform (CWT) to determine spatiotemporal characteristics, detrended fluctuation analysis (DFA) to examine gait complexity and autocorrelation analyses (ACF) to assess gait regularity. Reliability was examined using repeated running efforts and intraclass correlation. Proof of concept was determined by examining differences in each variable between various running speeds. Applicability was established by examining gait before and after fatiguing activity. The results showed most variables had excellent reliability. Test-retest R2 values for these variables ranged from 0.8 to 1.0. Low reliability was seen in bilateral comparisons of gait symmetry. Increases in running speed resulted in expected changes in spatiotemporal and acceleration variables. Fatiguing exercise had minimal effects on spatiotemporal variables but resulted in noticeable declines in complexity. This investigation shows that GPS-embedded trunk-mounted accelerometers can be effectively used to assess running gait. CWT and DFA yield reliable measures of spatiotemporal characteristics of gait and gait complexity. The effects of running speed and fatigue on these variables provides proof of concepts and applicability for this analytical approach. Master of Science Fitness trackers have become widely accessible and easy to use. So much so that athletic teams have been using them to track activity throughout the season. Researchers are able to manipulate data generated from the fitness monitors to assess many different variables including gait. Monitoring gait may generate important information about the condition of the individual. As a person fatigues, running form is theorized to breakdown, which increases injury risk. Therefore the ability to monitor gait may be advantageous in preventing injury. The purpose of this study is to show that the methods in this study are reproducible, respond reasonably to changes in speed, and to observe the changes of gait in the presence of fatigue or on tired legs. Three analyses are used in this study. The first method called autocorrelation, overlays acceleration signals of consecutive foot strikes, and determines the similarity between them. The second method utilizes a wave transformation technique that is able to determine foot contact times. The final method attempts to determine any pattern in the running stride. This method looks for changes in the structure of the pattern. Less structure would indicate a stride that is fatigued. The results showed that the methods of gait analysis used in this study were reproducible and responded appropriately with changes in speed. Small changes in gait were observed due to the presence of fatigue. Further investigation into the use of these methods to determine changes in gait due to the presence of fatigue are warranted.

Background To describe the identification, adaptation, and testing of an evidence-based pediatric weight management program for a health disparate community. Methods A community advisory board (CAB) of decision-makers and staff from local health care, public health, and recreation organizations engaged with academic partners to select an evidence-based program (EBP) for local implementation. Three EBPs were identified (Traffic Light, Bright Bodies, Golan and colleagues Home Environmental Model) and each EBP was rated on program characteristics, implementation and adaptation, and adoptability. Following selection of the EBP that was rated highest, the POPS-CAB made adaptations based on the program principles described in peer-reviewed publications. The adapted intervention, iChoose, was then pilot tested in 3 iterative phases delivered initially by research partners, then co-delivered by research and community partners, then delivered by community partners. The RE-AIM framework was used to plan and evaluate the iChoose intervention across all waves with assessments at baseline, post program (3 months), and follow-up (6 months). Results Bright Bodies rated highest on program characteristics and adoptability (p’s

Background SIPsmartER is a 6-month evidenced-based, multi-component behavioral intervention that targets sugar-sweetened beverages among adults. It consists of three in-person group classes, one teach-back call, and 11 automated phone calls. Given SIPsmartER’s previously demonstrated effectiveness, understanding its adoption, implementation, and potential for integration within a system that reaches health disparate communities is important to enhance its public health impact. During this pilot dissemination and implementation trial, SIPsmartER was delivered by trained staff from local health districts (delivery agents) in rural, Appalachian Virginia. SIPsmartER’s execution was supported by consultee-centered implementation strategies. Methods In this mixed-methods process evaluation, adoption and implementation indicators of the program and its implementation strategy (e.g., fidelity, feasibility, appropriateness, acceptability) were measured using tracking logs, delivery agent surveys and interviews, and fidelity checklists. Quantitative data were analyzed with descriptive statistics. Qualitative data were inductively coded. Results Delivery agents implemented SIPsmartER to the expected number of cohorts (n = 12), recruited 89% of cohorts, and taught 86% of expected small group classes with > 90% fidelity. The planned implementation strategies were also executed with high fidelity. Delivery agents completing the two-day training, pre-lesson meetings, fidelity checklists, and post-lesson meetings at rates of 86, 75, 100, and 100%, respectively. Additionally, delivery agents completed 5% (n = 3 of 66) and 10% (n = 6 of 59) of teach-back and missed class calls, respectively. On survey items using 6-point scales, delivery agents reported, on average, higher feasibility, appropriateness, and acceptability related to delivering the group classes (range 4.3 to 5.6) than executing missed class and teach-back calls (range 2.6 to 4.6). They also, on average, found the implementation strategy activities to be helpful (range 4.9 to 6.0). Delivery agents identified strengths and weakness related to recruitment, lesson delivery, call completion, and the implementation strategy. Conclusions In-person classes and the consultee-centered implementation strategies were viewed as acceptable, appropriate, and feasible and were executed with high fidelity. However, implementation outcomes for teach-back and missed class calls and recruitment were not as strong. Findings will inform the future full-scale dissemination and implementation of SIPsmartER, as well as other evidence-based interventions, into rural health districts as a means to improve population health.

Introduction: A number of effective physical activity programs for older adults exist, but are not widely delivered within community settings, such as the Cooperative Extension System. The purpose of this paper was to determine if an evidence-based intervention (EBI) developed in one state Extension system could be scaled-out to a new state system. Methods and results: The RE-AIM (reach, effectiveness, adoption, implementation, maintenance) framework was used to guide an iterative evaluation of three translational stages. Stage 1: Before program adoption, Extension health educators were surveyed and interviewed to assess physical activity programming perceptions and factors that may influence their decision to attend training or deliver the program in practice. Results indicated that a virtual, scalable training protocol would be necessary and that training needed to include hands-on instruction and be catered to those who were less confident in physical activity program delivery. Stage 2: Training attendees were surveyed pre- and post-training on factors related to the adoption-decision making process and contacted post-training to assess program delivery status. Training did not influence perceptions of the program, intent to deliver, or confidence in delivering the program. Stage 3: During program implementation, the program was evaluated through the RE-AIM framework by surveying across three key stakeholder groups: (1) program participants, (2) potential delivery personnel, and (3) Extension administrators. Findings indicate that the program has the potential to reach a large and representative proportion of the target audience, especially in rural areas. However, adoption and implementation rates among Extension health educators and community partners were low and data collection for effectiveness, implementation, and maintenance was a challenge. Conclusion: Overall, the results indicate initial struggles to translating and evaluating the program in a large, rural state. Implications for practice include making system-level changes to increase physical activity program adoption rates among Extension health educators and improve data collection and program evaluation through this community-based organization. More work is needed to identify infrastructure support and capacity to scale-out EBIs.

Cytosolic and mitochondrial human branched chain aminotransferase (hBCATc and hBCATm, respectively) play an integral role in brain glutamate metabolism. Regional increased levels of hBCATc in the CA1 and CA4 region of Alzheimer's disease (AD) brain together with increased levels of hBCATm in frontal and temporal cortex of AD brains, suggest a role for these proteins in glutamate excitotoxicity. Glutamate toxicity is a key pathogenic feature of several neurological disorders including epilepsy associated dementia, AD, vascular dementia (VaD) and dementia with Lewy bodies (DLB). To further understand if these increases are specific to AD, the expression profiles of hBCATc and hBCATm were examined in other forms of dementia including DLB and VaD. Similar to AD, levels of hBCATm were significantly increased in the frontal and temporal cortex of VaD cases and in frontal cortex of DLB cases compared to controls, however there were no observed differences in hBCATc between groups in these areas. Moreover, multiple forms of hBCATm were observed that were particular to the disease state relative to matched controls. Real-time PCR revealed similar expression of hBCATm mRNA in frontal and temporal cortex for all cohort comparisons, whereas hBCATc mRNA expression was significantly increased in VaD cases compared to controls. Collectively our results suggest that hBCATm protein expression is significantly increased in the brains of DLB and VaD cases, similar to those reported in AD brain. These findings indicate a more global response to altered glutamate metabolism and suggest common metabolic responses that might reflect shared neurodegenerative mechanisms across several forms of dementia. Alzheimer's Research Trust, UK (ARUK); BRACE (Bristol Research into Alzheimer's and Care of the Elderly); Brains for Dementia Research; Medical Research Council This work was supported by Alzheimer's Research Trust, UK (ARUK). The SWDBB, which provided the tissue and clinical data for this study, is supported by BRACE (Bristol Research into Alzheimer's and Care of the Elderly), Brains for Dementia Research and the Medical Research Council. We would like to also acknowledge the valuable academic input from Prof Seth Love, University of Bristol, UK.

Many community-based lifestyle interventions targeting African Americans have reported positive effects on participants' dietary choices and physical activity habits. However, these effects vary and not all participants will have outcome changes. Moderation analysis can help explain differential effects observed, but are not often reported. Hence, the objective of this secondary analysis was to explore potential moderators of intervention dose effects on diet quality and physical activity outcomes in an effective lifestyle intervention. Delta Body and Soul III, conducted from 2011 to 2012, was a 6-month, church-based, multicomponent, educational intervention designed to improve diet quality and increase physical activity in rural Southern African American adults. Generalized linear mixed models were used to determine associations among indicators of intervention dose received by participants, potential moderators and health outcome changes. Results indicated only three baseline characteristics-employment status, food shopping frequency and individual with primary responsibility for meal preparation-moderated the effects of education session attendance on diet quality changes. No evidence for moderation of exercise class attendance effects on physical activity changes was found. Thus, this culturally targeted, multicomponent lifestyle intervention did induce positive health changes in participants with a range of sociodemographic characteristics and food shopping and eating behaviors. US Department of Agriculture, Agricultural Research ServiceUnited States Department of Agriculture (USDA) [6401-51000-001-00D]; US Department of Health and Human Services, Health Resources Services Administration [6 U1FRHA07411] This work was supported by the US Department of Agriculture, Agricultural Research Service (project 6401-51000-001-00D); and the US Department of Health and Human Services, Health Resources Services Administration (6 U1FRHA07411). The views expressed are solely those of the authors' and do not reflect the official policy or position of the US government. Public domain – authored by a U.S. government employee

Overactive mitochondrial fission was shown to promote cell transformation and tumor growth. It remains elusive how mitochondrial quality is regulated in such conditions. Here, we show that upregulation of mitochondrial fission protein, dynamin related protein-1 (Drp1), was accompanied with increased mitochondrial biogenesis markers (PGC1α, NRF1, and Tfam) in breast cancer cells. However, mitochondrial number was reduced, which was associated with lower mitochondrial oxidative capacity in breast cancer cells. This contrast might be owing to enhanced mitochondrial turnover through autophagy, because an increased population of autophagic vacuoles engulfing mitochondria was observed in the cancer cells. Consistently, BNIP3 (a mitochondrial autophagy marker) and autophagic flux were significantly upregulated, indicative of augmented mitochondrial autophagy (mitophagy). The upregulation of Drp1 and BNIP3 was also observed in vivo (human breast carcinomas). Importantly, inhibition of Drp1 significantly suppressed mitochondrial autophagy, metabolic reprogramming, and cancer cell viability. Together, this study reveals coordinated increase of mitochondrial biogenesis and mitophagy in which Drp1 plays a central role regulating breast cancer cell metabolism and survival. Given the emerging evidence of PGC1αcontributing to tumor growth, it will be of critical importance to target both mitochondrial biogenesis and mitophagy for effective cancer therapeutics.

Non-nutritive sweeteners (NNS) are frequently used as substitutes for added sugars. NNS are difficult to study due to the inability to accurately measure the amounts individuals consume, as well as limitations in study design and methods, including reliance on observational study designs, the use of diet soda as a proxy of total NNS intake, and the grouping of NNS into a single category rather than studying NNS as individual products. New dietary assessment methods and improved study design and methods are needed to advance researchers' abilities to study NNS and their impact on consumers' health. The objectives of this dissertation were to 1) determine validity and reproducibility of a novel NNS food frequency questionnaire (NNS-FFQ), 2) develop methodology for an objective NNS urinary biomarker, 3) identify an appropriate carrier for NNS intake in studies, and 4) examine the literature on the relationship between NNS and weight-related outcomes based on study design and methods. Objectives 1 and 2: participants (n=125) completed three 24-hr dietary recalls, the NNS-FFQ, and 2 24-hr urine samples. NNS intake via NNS-FFQ and recalls were compared using Bland-Altman analyses, with agreement levels ranging from 92.7-99.2% for individual NNS types and total intake. The NNS biomarker methodology was developed using ultra performance liquid chromatography (UPLC-MS/MS), which analyzes each sample for the presence of NNS and related metabolites. This method observed a range from very strong presence of NNS to not detectable, indicating that this biomarker could identify specific NNS consumption (n=9). Objective 3: a sensory evaluation (n=67) was conducted to identify if applesauce or water was a more appropriate carrier for NNS for future interventions. Applesauce was preferred (sucralose=83.6%; aspartame=79.1%; stevia=74.6%) significantly more than water (p≤0.001), indicating that applesauce could be used as an acceptable carrier of NNS in research studies. Objective 4: a systematic literature review focusing on study design and methods used in investigations on NNS and weight-related outcomes found that 81% of RCT had improved weight outcomes, while 76% of observational studies had higher weight outcomes. Improving NNS study design and methods will increase the quality of research conducted on NNS and related health outcomes. Ph. D. Artificial sweeteners [non-nutritive sweeteners (NNS)] are often used to replace added sugars. NNS are difficult to study due to the inability to accurately measure the amounts individuals consume, as well as limitations in study design and methods, including reliance on observational study designs, the use of diet soda to represent total NNS intake, and the grouping of NNS into a single category rather than studying NNS as individual products. New dietary assessment tools and improved study design and methods are needed to allow researchers better to study NNS and their impact on health. The objectives of this dissertation were to 1) to determine the ability of a NNS food frequency questionnaire (NNS-FFQ) to measure typical NNS intake, 2) develop methodology for an objective NNS urinary biomarker, 3) identify an appropriate carrier for NNS intake in studies, and 4) to review the currently available research on the relationship between NNS and weight status. Objectives 1 and 2: 125 participants completed three 24-hr dietary recalls, the NNS-FFQ to measure usual NNS intake, and 24-hour urine samples for a NNS biomarker. Amounts of NNS that consumers reported in recalls were compared to amounts reported in the NNS-FFQ, with the tools finding similar NNS amounts in participants’ diets. The NNS biomarker methodology was developed using ultra performance liquid chromatography (UPLC-MS/MS). This rapid method measures the presence of NNS and related products (saccharin, acesulfame potassium, sucralose, steviol glucuronide, and erythritol) in urine. Among 9 participants, this method identified wide differences, ranging from strong presence of NNS to not measureable. Objective 3: participants (n=67) completed a sensory evaluation (taste testing) study to identify a more appropriate food or beverage (applesauce or water) for NNS (sucralose, aspartame, and stevia) to be mixed with in research studies. Applesauce was preferred over water for all sweeteners (>74%), indicating that applesauce would be an acceptable NNS carrier. Objective 4: a systematic review on study design and research methods used in studies on NNS and weight-related outcomes. Improving NNS measurement tools and study design methods will improve the quality of research that can be conducted on NNS and related health outcomes.

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, 0.05) but decreased in the sedentary (34 ± 7% to 4 ± 5% and 78 ± 26% to -21 ± 6%, respectively; both P0.05). 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.

Skeletal muscle is highly involved in macronutrient metabolism. To maintain proper energy metabolism and physiology, skeletal muscle must adapt to nutrient supply. Thus, diet macronutrient composition is an important modulator of skeletal muscle metabolism. Evidence from rodent and human models show high-fat diets contribute to impaired insulin signaling, as well as decreased fatty acid and glucose oxidation. Utilizing proteomic analysis of metabolic proteins in humans may lead to the mechanism behind skeletal muscle adaption to macronutrient composition, potentially providing the groundwork for characterizing the etiology of high-fat feeding induced metabolic disease. The objective of this study was to compare the substrate oxidation patterns and the levels of metabolic proteins in the fasted skeletal muscle of lean, healthy males that either increased fatty acid oxidation in response to the high-fat diet, termed responders, or males that decreased fatty acid oxidation, termed non-responders. We employed a controlled feeding study design, where the participants served as their own controls. Following a 2-week control diet (30% fat, 55% carbohydrate and 15% protein), participants came to the lab fasted overnight and a muscle biopsy was taken from their vastus lateralis muscle. Participants were then placed on a 5-day high-fat diet (50% fat [45% saturated fat], 35% carbohydrate, and 15% protein). Following this diet, participants again came to the lab fasted overnight and another muscle biopsy was taken from their vastus lateralis muscle. Both the control and the high-fat diets were isocaloric to habitual diets. Muscle from the biopsies were utilized for substrate metabolism measures and mass spectrometry. We did not observe any significant differences in glucose oxidation between responders and non-responders, prior to or following the high-fat diet. Our proteomic analysis identified 81 proteins and protein subunits involved in substrate metabolism but only 6 were differentially regulated by the high-fat diet. Independent of the high-fat diet, compared to non-responders, responders contained an overall higher content of protein subunits belonging to Complex I and ATP synthase. The findings from this study suggest that adaption to high-fat feeding is individual specific and proteomic changes alone cannot explain high-fat feeding induced metabolic changes. Ph. D. Skeletal muscle is highly involved in macronutrient metabolism, which consist of the breakdown and utilization of glucose and fatty acids, thus making the foods we ingest a major modulator of skeletal muscle metabolism. Over the last few decades, Americans have increased their ingestion of foods high in saturated fats, which has coincided with the increased prevalence of obesity and type 2 diabetes. Further, evidence suggests these metabolic diseases are associated with the skeletal muscle’s inability to switch between the utilization of glucose and fatty acid in response to nutrient supply. Analyzing metabolic protein content in humans may lead to the mechanism behind skeletal muscle adaption to macronutrient composition, potentially leading to the cause behind the development of high-fat feeding induced metabolic disease. The objective of our controlled feeding study was to compare the macronutrient metabolism and the content of metabolic proteins in the fasted skeletal muscle of healthy males that either increased fatty acid utilization in response to a high-fat diet, termed responders, or males that decreased fatty acid utilization, termed non-responders. Following a 2-week control diet (30% fat, 55% carbohydrate and 15% protein), participants came to the lab fasted overnight and a biopsy was taken from their thigh muscle called the vastus lateralis. Participants then began a 5-day high-fat diet (50% fat [45% saturated fat], 35% carbohydrate, and 15% protein). Following this diet, participants came to the lab fasted overnight and another biopsy was taken from their vastus lateralis muscle. Both the control and the high-fat diets were isocaloric to habitual diets. The muscle samples were used to analyze macronutrient metabolism and identify metabolic protein content. We did not observe differences in glucose utilization between responders and non-responders, prior to or following the high-fat diet. We identified 81 metabolic proteins and protein subunits but only 6 were differentially regulated by the high-fat diet. Independent of diet, responders contained higher levels of subunits from 2 proteins involved in cell energy production, Complex I and ATP synthase. Our findings suggest that adaption to high-fat feeding is individual specific and protein content changes alone cannot explain high-fat feeding induced metabolic changes.