Chronic low-grade inflammation is a main pathogenic link between obesity and Type 2 Diabetes (T2D) and a putative target for treatment. While a wide array of pharmacologic agents is available to manage T2D, many patients still face perturbed glycemia and subclinical inflammation. Therefore, complementary nutraceutical strategies that target inflammation, metabolism, and resolution physiology hold promise as adjunctive options to quell the disturbed immuno-metabolic milieu observed in T2D. Omega-3 polyunsaturated fatty acids (PUFAs) and anthocyanins are two dietary components evidenced to mitigate inflammation and improve T2D risk factors, through distinct and similar targets. However, the combined use of such nutraceuticals has not yet been examined in individuals with T2D. This dissertation leveraged data from a larger randomized, double-blind, placebo-controlled trial conducted between January 2022—September 2023 investigating the use of combined supplementation (active treatment; [FOM]) of anthocyanins (600 mg/d maqui berry extract) and omega-3 PUFAs (3 g/day fish oil; 2 g/d EPA, 1 g/d DHA) for 8 weeks on cytokines and mental acuity in individuals with T2D, compared to a placebo (CON). The current study examined the effects of this supplemental strategy on markers of metabolic inflammation, oxidative stress, and cardiometabolic risk. The results indicated that a marker of sustained omega-3 dietary intake and tissue accumulation termed the Omega-3 Index was inversely associated with HbA1c (? = -8.5, 95%CI -15.1, -1.4, p = 0.022) and glucose (? = -12.4, 95%CI -22.9, -0.5, p = 0.042), after adjustment for covariates at baseline across all participants with T2D in this study. However, outcomes from linear mixed model analyses demonstrated that there were no significant differences in change from baseline between FOM and CON groups at week 8 in any of the inflammatory, oxidative stress, glycemic control, or circulating lipid markers assessed in this study. These null effects were observed despite a 93% greater increase from baseline in the Omega-3 Index observed in the FOM group compared to the CON group at week 8. Therefore, the findings do not support significant treatment effects associated with 2 months of combined marine omega-3 PUFAs and maqui berry extract on inflammatory and cardiometabolic outcomes in individuals with T2D.

Metabolomics focuses on the study of metabolic changes occurring in varioussystems and utilizes quantitative and semi-quantitative measurements of multiple metabolites in parallel. Mass spectrometry (MS) is the most ubiquitous platform in this field, as it provides superior sensitivity regarding measurements of complex metabolic profiles in biological systems. When combined with MS, multivariate statistics and advanced machine learning algorithms provide myriad opportunities for bioinformatics insights beyond simple univariate data comparisons. In this dissertation, the application of MS-based metabolomics is introduced with an emphasis on biomarker discovery for human disease detection. To advance disease diagnosis using MS-based metabolomics, numerous statistical techniques have been implemented in this research including principal component analysis, factor analysis, partial least squares-discriminant analysis (PLS-DA), orthogonal PLS-DA, random forest, receiver operating characteristic analysis, as well as functional pathway/enzyme enrichment analyses. These approaches are highly useful for improving classification sensitivity and specificity related to disease-induced biological variation and can help identify useful biomarkers and potential therapeutic targets. It is also shown that MS-based metabolomics can distinguish between clinical and prodromal disease as well as similar diseases with related symptoms, which may assist in clinical staging and differential diagnosis, respectively. Additionally, MS-based metabolomics is shown to be promising for the early and accurate detection of diseases, thereby improving patient outcomes, and advancing clinical care. Herein, the application of MS methods and chemometric statistics to the diagnosis of breast cancer, coccidioidomycosis (Valley fever), and senile dementia (Alzheimer's disease) are presented and discussed. In addition to presenting original research, previous efforts in biomarker discovery will be synthesized and appraised. A Comment will be offered regarding the state of the science, specifically addressing the inefficient model of repetitive biomarker discovery and the need for increased translational efforts necessary to consolidate metabolomics findings and formalize purported metabolic markers as laboratory developed tests. Various factors impeding the translational throughput of metabolomics findings will be carefully considered with respect to study design, statistical analysis, and regulation of biomedical diagnostics. Importantly, this dissertation will offer critical insights to advance metabolomics from a scientific field to a practical one including targeted detection, enhanced quantitation, and direct-to-consumer considerations.

Oral contraceptives are one of the most frequently used forms of birth control among young women. However, research has shown that this type of medication can contribute to negative changes in mood and diminished vitamin status. In particular, women taking oral contraceptives are at an increased risk of vitamin B6 deficiency due to changes in enzyme activity with estrogen intake. Depressed mood is one of the known symptoms of vitamin B6 deficiency as this vitamin acts as an essential cofactor in converting tryptophan to the neurotransmitter, serotonin. Lack of adequate levels of vitamin B6 therefore contribute to decreased production of serotonin and subsequent changes in mood, including symptoms of depression. With vitamin B6 being the most common nutrient deficiency, and the ever increasing prevalence of depression in the United States, especially among young adults, it is crucial that researchers investigate ways to mitigate both of these undesirable side effects. Current research on the topic fails to directly connect supplementation of vitamin B6 to positive changes in mood in oral contraceptive users.

This 12-week long double-blinded, placebo-controlled crossover trial examined the effects of daily supplementation of vitamin B6 as 100 mg of pyridoxine hydrochloride, on mood states in 8 healthy college women (18-25 y) that use combined oral contraceptives. Vitamin status was assessed via plasma pyridoxal 5’-phosphate (PLP). Plasma PLP levels significantly increased by >193% (p=0.003) with daily supplementation of 100 mg B6 over a four week period. Mood changes with supplementation were assessed using the Profile of Mood States (POMS). Although a small improvement in the POMS depression sub score was observed after 4 weeks of vitamin B6 supplementation (14.7%), the changes were insignificant (p>0.05). Furthermore, total mood disturbance scores did not significantly change with either the placebo or supplement periods. While mood states were not improved, a significant decrease in the presence of depressive symptoms as measured by the Beck Depression Inventory was observed after vitamin B6 supplementation, compared to placebo (p=0.047). The results of this study necessitate further investigation into the use of B6 supplementation as a means of reducing negative mood changes in oral contraceptive users.

According to a 2016 census, eight million adults conform to a vegetarian diet within the United States, and about 50% of these adults follow a vegan diet. The census determined that plant-based diets are quickly growing in popularity particularly in young adults between the ages of 18 to 34 years. Many Americans are aware of the health benefits of a plant-based diet, however, the dietary risks associated with these diets are not well emphasized. Health concerns such as vitamin deficiencies and altered metabolism are heightened in vegetarian populations.

One Particular nutrient that is commonly lacking in the vegetarian diet is vitamin B12. Vitamin B12 is found mainly in animal-derived food sources such as meat, poultry, fish, dairy, and eggs. Although some vegetarians, called lacto-ovo vegetarians, consume dairy and eggs, vegans do not consume any animal products at all. Vitamin B12 deficiency can have devastating consequences on the human body due to its role as a methylation cofactor. Metabolism, DNA replication, and cancer formation all involve methylation processes.

This cross-sectional, differential study aimed to further understand the relationship between vegetarianism, vitamin B12 status, and methylation capacity in healthy adults. A group of 34 healthy adults (18 vegetarians and 16 omnivores) was recruited to analyze serum B12, homocysteine, methylmalonic acid, serum total folate, and transcobalamin II status. It was hypothesized that (1) vegetarians would have a lower vitamin B12 status, and thus, a lower methylation capacity than omnivores and that (2) low vitamin B12 status would be correlated with low methylation capacity.

The data show that vegetarians did not have significantly lower vitamin B12 methylation capacity status than omnivores. Nor was vitamin B12 status correlated with methylation capacity. However, the data revealed that diet quality had a positive influence on folate status. There was also a statistical trend (p=0.08) for homocysteine reduction in participants consuming high-quality diets. The data herein suggest that methylation capacity may be impacted by the quality of diet rather than the type of diet.

Obesity prevalence is high in the United States, in part due to increased fat storage following consumption of high fat/carbohydrate (sugar) foods. Following a meal, carbohydrate stimulates its own oxidation, while simultaneously suppressing fat oxidation, ultimately leading to fat storage. Aerobic exercise preceding a meal increases fat oxidation in the postprandial period, which may reduce fat storage. The ideal exercise prescription for optimal postprandial fat oxidation is unknown. The effect of low and moderate intensity continuous exercise (MIE) has been studied extensively, while the effects of high-intensity interval exercise (HIIE) on post-prandial substrate oxidation has not been examined. The purpose of this study was to compare the effects of MIE and HIIE on postprandial substrate oxidation after consumption of an isocaloric meal (2 glazed donuts; 520 kcal) in healthy adults. Ten subjects (8 males, 2 females; age=24yr, BMI=24 kg/m2) completed three conditions in random order: 1) no exercise control; 2) MIE: cycling at 60-75%HRmax; 3) HIIE: cycling at 90-95%HRmax. The duration of each exercise bout was sufficient to expend approximately 520 kcal, the energy equivalent of the donuts, which were consumed 1 hour post-exercise. Immediately after consuming the donuts, pulmonary ventilation and gas exchange were measured breath-by-breath continuously and recorded (min-by-min) for 5 hours. Repeated measures analysis of covariance was used to compare the mean differences in outcome variables accounting for gender. Absolute postprandial fat oxidation (g/5 hours) was 17.3±5.4, 27.1±9.6 and 23±1.2 for control, MIE and HIIE trials respectively, with the postprandial fat oxidation significantly greater for the two exercise conditions compared to control. Relative to baseline values, both exercise conditions resulted in cumulative net postprandial fat oxidation significantly greater than control (control = -1.79±3.99g; MIE = 11.51±8.41g, HIIE= 9.51±5.20g). Therefore, results indicate that exercise most certainly increases postprandial fat oxidation, and that exercise type, either MIE or HIIE, is not as important as total energy expended. The fact that exercise of ~1 hour was required to oxidize the amount of fat in two donuts, that required only a few minutes to consume, highlights the challenges of using exercise for weight control in an obesogenic environment.

ABSTRACT

Objective: The purpose of this randomized parallel two-arm trial was to examine the effect that an intervention of combining daily almond consumption (2.5 ounces) with a walking program would have on heart rate recovery and resting heart rate when compared to the control group that consumed a placebo (cookie butter) in men and postmenopausal women, aged 20-69, in Phoenix, Arizona.

Design: 12 men and women from Phoenix, Arizona completed an 8-week walking study (step goal: 10,000 steps per day). Subjects were healthy yet sedentary, non-smokers, free from gluten or nut allergies, who had controlled blood pressure. At week 5, participants were randomized into one of two groups: ALM (2.5 oz of almonds daily for last 3 weeks of trial) or CON (4 tbsp of cookie butter daily for last 3 weeks of trial). Body weight, BMI, and percent body fat were measured using a stadiometer and Tanita at the screening visit. Resting heart rate, heart rate recovery, and anthropometric measurements were taken at weeks 0, 5, and 8.

Results: 8 weeks of walking 10,000 steps per day, with or without 3 weeks of almond consumption did not significantly improve heart rate recovery (p=0.818) or resting heart rate (0.968).

Conclusions: Almond consumption in combination with a walking intervention does not significantly improve heart rate recovery or resting heart rate.

The stadiometer is the gold standard human height measure, but recent studies have begun to question whether laser technology is a better tool to measure height. The purpose of this study was to investigate if the laser device has inter-rater reliability, how the laser-device measures supine height in comparison to standard methods, and if the laser device will be consistent in measuring human height shorter, as seen in previous studies. Two investigators measured a total of 80 adults independently. Measurements included knee height, arm span, demi span, supine height by laser, standing height by laser and standing height by stadiometer. There was a strong inter-rater reliability for the laser height measurement: excluding one outlier r=0.998. Supine height measures done with a laser were strongly correlated with arm span, but mean values were closest between supine height and knee height (171.3cm and 171.2cm). The laser measured standing height 0.5cm shorter, on average, than the stadiometer. It is concluded that the laser device is a reliable, validated tool to measure human height, standing or supine.

Background: Despite the reported improvements in glucose regulation associated with flaxseeds (Linum usitatissimum) few clinical trials have been conducted in diabetic participants. Objective: To evaluate the efficacy of ground flaxseed consumption at attenuating hyperglycemia, dyslipidemia, inflammation, and oxidative stress as compared to a control in adults with non-insulin dependent type 2 diabetes (T2D). Design: In a randomized parallel arm controlled efficacy trial, participants were asked to consume either 28 g/d ground flaxseed or the fiber-matched control (9 g/d ground psyllium husk) for 8 weeks. The study included 17 adults (9 male, 8 females; 46±14 y; BMI: 31.4±5.7 kg/m2) with a diagnosis of T2D ≥ 6 months. Main outcomes measured included: glycemic control (HbA1c, fasting plasma glucose, fasting serum insulin, and HOMA-IR), lipid profile (total cholesterol, LDL-C, HDL-C, total triglycerides, and calculated VLDL-C), markers of inflammation and oxidative stress (TNF-alpha, TBARS, and NOx), and dietary intake (energy, total fat, total fiber, sodium). Absolute net change for measured variables (week 8 values minus baseline values) were compared using Mann-Whitney U non-parametric tests, significance was determined at p ≤ 0.05. Results: There were no significant changes between groups from baseline to week 8 in any outcome measure of nutrient intake, body composition, glucose control, or lipid concentrations. There was a modest decrease in TNF-alpha in the flaxseed group as compared to the control (p = 0.06) as well as a mild decrease in TBARS in the flaxseed as compared to the control group (p = 0.083), though neither were significant. Conclusions: The current study did not detect a measurable association between 28 g/d flaxseed consumption for 8 weeks in T2D participants and improvements in glycemic control or lipid profiles. There was a modest, albeit insignificant, decrease in markers of inflammation and oxidative stress in the flaxseed group as compared to the control, which warrants further study.

Background: Postprandial hyperglycemia can increase levels of oxidative stress and is an independent risk factor for complications associated with type 2 diabetes.

Purpose: To evaluate the acute effects of a 15-min postmeal walk on glucose control and markers of oxidative stress following a high-carbohydrate meal.

Methods: Ten obese subjects (55.0 ± 10.0 yrs) with impaired fasting glucose (107.1 ± 9.0 mg/dL) participated in this repeated measures trial. Subjects arrived at the laboratory following an overnight fast and underwent one of three conditions: 1) Test meal with no walking or fiber (CON), 2) Test meal with 10g fiber and no walking (FIB), 3) Test meal with no fiber followed by a 15-min treadmill walk at preferred walking speed (WALK). Blood samples were taken over four hours and assayed for glucose, insulin, thiobarbituric reactive substances (TBARS), catalase, uric acid, and total antioxidant capacity (TAC). A repeated measures ANOVA was used to compare mean differences for all outcome variables.

Results: The 2hr and 4hr incremental area under the curve (iAUC) for glucose was lower in both FIB (2hr: -93.59 mmol∙120 min∙L-1, p = 0.006; 4hr: -92.59 mmol∙240 min∙L-1; p = 0.041) and WALK (2hr: -77.21 mmol∙120 min∙L-1, p = 0.002; 4hr: -102.94 mmol∙240 min∙L-1; p = 0.005) conditions respectively, compared with CON. There were no differences in 2hr or 4hr iAUC for glucose between FIB and WALK (2hr: p = 0.493; 4hr: p = 0.783). The 2hr iAUC for insulin was significantly lower in both FIB (-37.15 μU ∙h/mL; p = 0.021) and WALK (-66.35 μU ∙h/mL; p < 0.001) conditions, compared with CON, and was significantly lower in the WALK (-29.2 μU ∙h/mL; p = 0.049) condition, compared with FIB. The 4hr iAUC for insulin in the WALK condition was significantly lower than both CON (-104.51 μU ∙h/mL; p = 0.001) and FIB (-77.12 μU ∙h/mL; p = 0.006) conditions. Markers of oxidative stress were not significantly different between conditions.

Conclusion: A moderate 15-minute postmeal walk is an effective strategy to reduce postprandial hyperglycemia. However, it is unclear if this attenuation could lead to improvements in postprandial oxidative stress.

Background Hemodialysis (HD) patients elicit an oxidant-antioxidant imbalance in addition to a selenium deficiency, possibly contributing to cardiovascular disease (CVD) mortality. Objective To evaluate the effect of selenium supplementation on CVD outcomes and antioxidant status in HD patients. Design A randomized controlled intervention trial conducted from October 2012 to January 2013. Participants/setting The study included 27 maintenance HD patients (61.1+17.5y, 14M, 13F) receiving HD in the greater Phoenix, AZ area. Intervention Patients received one of three treatments daily: 2 Brazil nuts, (5g, 181µg/day of selenium as selenomethionine [predicted]), 1 tablet of selenium (200µg/day of selenium as selenomethionine), or control (3 gummy bears). Main outcome measures Antioxidant status outcome measures included total antioxidant capacity, vitamin C, and RBC and plasma glutathione peroxidase (GSH-Px). CVD outcomes measures included brain natriuretic peptide; plasma cholesterol, high density lipoprotein, low density lipoprotein, triglycerides; blood pressure, and thoracic cavity fluid accumulation. Statistical analyses performed Repeated measures ANOVA analyzed changes over time and between groups at months 0 and 2 and months 0 and 3. Results Independent analysis showed the Brazil nuts provided 11µg of selenium/day and the pill provided 266µg of selenium/day. Consequently, the Brazil nut group was combined with the placebo group. 21 patients completed 2 months of the study and 17 patients completed the study in its entirety. Data was analyzed for months 0, 1 and 2. No significant differences were noted for antioxidant status outcome measures with the exception of plasma GSH-Px. Patients receiving the selenium pill had a significant increase in plasma GSH-Px compared to the placebo group (6.0+11 and -4.0+7.6, respectively, p=0.023 for change between month 0 and month 2). No significant differences were seen in total antioxidant capacity or for CVD outcome measures over time or between groups. Conclusions These data indicate that selenium supplementation increased plasma GSH-Px concentration in HD patients; however, oxidative stress was not altered by selenium supplementation. The low vitamin C status of HD patients warrants further research, specifically in conjunction with selenium supplementation.