Background: Nighttime eating is often associated with metabolic syndrome and poor body composition and these conditions may be influenced by the natural decline in metabolism that occurs during sleep. However, only limited research has been conducted to determine the role of individual macronutrients at night. Previous research indicates that protein consumption increases metabolic rate more than carbohydrates or fat, and therefore may attenuate this decline when consumed at night before bed. In addition, digestion and absorption kinetics of a fast protein such as whey protein (WP) and a slow protein such as casein protein (CP) may independently influence appetite and body composition. Therefore, nighttime eating may be a window of opportunity to influence changes in body composition, strength, cardiovascular health, metabolism and appetite (hunger, desire to eat, and satiety). Purpose: To compare the effects of isocaloric maltodextrin placebo (PLA), WP and CP supplements when consumed immediately prior to nocturnal sleep when combined with four weeks of exercise training on body composition, strength, fasting glucose and lipid profiles, metabolism and appetite. Methods: Fifty-nine sedentary, overweight and obese participants were recruited and had baseline measurements of body composition (dual energy X-ray absorptiometry (DXA)), resting metabolism (ParvoMedics TrueOne 2400 metabolic cart), strength (1RM Chest- and Leg-press), blood glucose and lipid profiles (Cholestech LDX Analyzer) and appetite questionnaires (visual analogue scale) taken after an overnight fast (0600-0900 h). Forty-eight participants completed the four-week study protocol. The participants were randomly stratified by % body fat, BMI, and gender to one of three groups: PLA (n= 14, men: 4, BMI= 34.4 ± 1.5 kg/m2, age= 28.1 ± 1.8 years), WP (n= 17, men: 3, BMI= 34.3 ± 1.3 kg/m2, age= 30.1 ± 1.6 years), CP (n=17, men: 3, BMI= 35.4 ± 1.3 kg/m2, age= 30.1 ± 1.6 years) in a double blind design. Participants were then instructed to consume their supplement at least two hours after dinner and no more than 30 minutes before bed each night for four weeks. All participants attended supervised exercise sessions (3x/week; 2 days of resistance exercise and 1 day of high-intensity cardiovascular exercise). Post-testing occurred 36-60 hours after the last supplementation and 96-144 hours after the final training session. A one-way ANOVA was performed to examine possible group differences at baseline and differences in change among groups. A two-way ANOVA with repeated measures was used to evaluate changes in dependent variables over time ([pre x post] x [PLA x WP x CP]). A Tukey test was used for post hoc comparisons. Values are reported as means ± SEM. Significance was accepted at P<0.05. Results: Eleven participants who completed baseline measurements failed to complete the four-week protocol and maintain satisfactory compliance with exercise and supplement intake (< 80% compliance). With the exception of fasting glucose, no significant group differences existed at baseline. There were no group x time interactions for resting metabolic rate (RMR), hunger, satiety, desire to eat, fat mass, lean body mass, BF%, or weight, although RMR displayed a trend (P=0.0559) towards significance with the PLA group decreasing by 74.3 ± 94.5 kcal/day and WP and CP increasing by 235.7 ± 84.5 kcal/day and 51.7 ± 79.4 kcal/day, respectively. Additionally, there was a group x time effect for VO2 with WP increasing by 0.3 ± 0.1 ml/kg/min compared with a decrease of 0.1 ± 0.1 ml/kg/min and an increase of 0.1 ± 0.1 ml/kg/min for PLA and CP, respectively. Significant time effects were measured for satiety (pre: 31.5 ± 2.3 mm, post: 40.6 ± 2.3 mm, P< 0.008) and lean body mass (LBM) (pre: 51.8 ± 0.1 kg, post: 52.3 ± 0.1 kg, P< 0.0001). Conclusion: In conclusion, our data indicate exercise three times per week for four weeks combined with nighttime eating can be a successful method for improving LBM, BF%, strength, and satiety in previously sedentary, overweight and obese individuals. Additionally, four weeks of nighttime WP supplementation may elevate VO2 36-60 hours after the last supplementation. / A Thesis submitted to the Department of Nutrition, Food & Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. / Summer Semester, 2012. / June 21, 2012. / Carbohydrate vs. protein at night, Casein, Digestion and Absorption kinetics, Nighttime, Protein, Whey / Includes bibliographical references. / Michael J. Ormsbee, Professor Directing Thesis; Lynn B. Panton, Committee Member; Robert J. Contreras, Outside Committee Member.

The consumption of whole foods has shown to offer more synergistic health benefits as compared to foods made with highly processed ingredients. The American food industry has a variety of food products that consist of refined, isolated or artificial ingredients (e.g. bread, beverages, snack foods and others). Although the sensory properties of several products are satisfactory, many consumers desire products made with minimally processed ingredients. The primary goal of the project was to develop a satisfactory alternative ice cream for an existing brand name ice cream using minimally processed sweetener, blue agave nectar, instead of sucrose that is commonly used in the brand name ice cream. The rheological properties for consistency, melting, texture (firmness) and surface appearance (color) were compared between the experimental and the control ice cream formulae. Five sets of data were collected for 2 different batches (10 data sets) of both formulae. The paired results indicated no significant difference in the melting rate (p = .25) between the experimental (16.15 ± 2.75) and controlled formulae (17.55 ± 1.28). The average rate of flow (consistency) was also similar (p = .95) between the experimental (4.48 ± 0.92) and the controlled (4.50 ± 0.65). However, there was a significant difference (p = .001) in the exertion of force. The experimental samples averaged to be softer in texture with a Mean force of 0.30 (± 0.16) as compared to the control samples (0.63 ± 0.20) with distance and time being constant. Also, the experimental samples, on an average, were more yellow (Y glossy = 1.09) than the control (Y glossy = 0.96) as determined by the reflectometer readings. Based on the rheological tests, the experimental ice cream appears to be of comparable quality to that of the same qualities of the control. Overall, the experimental ice cream may provide an acceptable alternative to the ice cream sweetened with sucrose. However, a human sensory analysis should be conducted to determine the overall desirability of the product. KEY WORDS: Ice cream, texture, color, rheological properties, agave / A Thesis submitted to the Department of Nutrition, Food, and Exercise Sciences in partial fulfillment of the requirements for the degree of Masters of Science. / Fall Semester, 2011. / November 3, 2011. / agave, color, ice cream, rheological properties, texture / Includes bibliographical references. / Shridhar K. Sathe, Professor Directing Thesis; Penny A. Ralston, Committee Member; Doris Abood, Committee Member.

Anxiety and depressive disorders are the most common of all psychiatric disorders; however, current human and animal research has yet to provide a clear understanding of the neural mechanisms underlying their etiology. Demographic analyses illustrate not only the enormous prevalence and incidence of these affective disorders, but also the pervasive gender discrepancy seen worldwide in patients suffering from anxiety and depression. There are largely documented sex differences in mood disorders, where females are more than twice as likely as men to be afflicted with depression and anxiety. Overall, sexually dimorphic aspects of anxiety and depression are well documented but poorly understood, warranting additional research delving into the mechanisms behind these sex differences. Considerable sex differences occur in the incidence and prevalence of anxiety disorders where women are more anxious than men, particularly in situations where social interaction is required. In preclinical studies, the social interaction test represents a valid animal model to study sex differences in social anxiety. Indeed, female rats engage less in conspecific interactions than their male counterparts, which are behaviors indicative of higher social anxiety in female rats. Given both the high prevalence of anxiety disorders in women and the fact that little is known about the mechanisms of gender differences in anxiety, our primary aim in our first study was to investigate the neurobiological mechanisms underlying sex differences in social anxiety-like behavior in rats. We investigated the activation of several brain areas using the neuronal marker zif268 and discovered sexually dimorphic zif268 expression, increased in the male, specifically within the medial prefrontal cortex (mPFC). Through the use of zif268 antisense oligodeoxynucleotides (zif ASO), we induced a temporary downregulation of zif268 expression in the mPFC of male and female rats and found that zif268 ASO male rats show more social anxiety-like behaviors when compared with control male rats in the social interaction test. In fact, zif268 ASO males displayed social anxiety-like behaviors, which were similar to control females. Thus, downregulation of zif268 expression in the mPFC of male rats eliminated sex differences previously found in the social anxiety-like behavior tests. Interestingly, zif268 ASO in female rats had no effect on their social interaction. In our second study, we investigated the role of extracellular signal regulated kinase 2 (ERK2), an upstream regulator of zif268, in the medial prefrontal cortex (mPFC). Indeed, female rats' had lower ERK2 expression compared to male rats, and overexpression of ERK2 in the mPFC increases their social interaction to the level xii seen in their male counterparts. Our novel findings have led us to ascertain that sexually dimorphic zif268 and ERK2 expression in the mPFC are key molecular factor in mediating sex-specific anxiety-like behavior in the social interaction test. Human and animal studies suggest that testosterone may have antidepressant effects. In our third study, we sought to investigate the molecular mechanisms underlying the antidepressant effects of testosterone within the hippocampus, an area that is fundamental in the etiology of depression. The effects of testosterone replacements in gonadectomized adult male rats were investigated using the sucrose preference and forced swim tests. We explored possible effects of testosterone on hippocampal neurogenesis and gene expression of stress-related molecules. Through the use of viral vectors, we pursued the antidepressant molecular mechanism(s) of testosterone in mediating anhedonia and manipulated ERK2 expression in the dentate gyrus in gonadectomized rats with testosterone replacements. Testosterone had antidepressant effects, likely mediated by aromatization to estrogen metabolites, in the sucrose preference and forced swim tests despite having no effects on hippocampal cell proliferation or survival. We found a testosterone-dependent regulation of hippocampal ERK2 expression. Functionally, reducing ERK2 activity within the dentate gyrus induced anhedonia in gonadectomized rats receiving testosterone supplementation, whereas the overexpression of ERK2 rescued this behavior in gonadectomized rats. These results implicate a role for ERK2 signaling within the dentate gyrus area of the hippocampus as a key mediator of the antidepressant effects of testosterone. In our fourth study, we investigated the antidepressant effects and interactions between testosterone and imipramine in both male and female rats subjected to stressful conditions in order to model a depressive-like state. A chronic social isolation model was used to induce an anxiety and depressive-like behaviors in adult gonadectomized male and ovariectomized female rats receiving chronic testosterone and imipramine treatments. Their anxiety and depression-like behaviors were examined using the light-dark box, elevated plus maze, open field, sucrose preference and novelty induced hypophagia tests. In socially isolated rats, the anxiolytic and antidepressant effects of testosterone and imipramine were limited to male rats. Additionally, testosterone enhanced the neurogenic effect of imipramine on hippocampal cell proliferation in male rats. Although female rats exhibited signs of anxiety and depressive-like behaviors following social isolation, testosterone and/or imipramine administration had no anxiolytic or xiii antidepressant effects in ovariectomized females. These results suggest that testosterone and imipramine had anxiolytic and antidepressant effects in socially isolated male, but not female rats, and that testosterone enhances the effect of imipramine on cell proliferation in the hippocampus of male rats only. These studies have investigated the mechanisms underlying sex differences in the incidence and prevalence of anxiety and depressive disorders. We concentrated on the influence of gonadal hormones and several molecular targets on anxiety and depressive-like behaviors in the Sprague-Dawley rat. Overall, these studies underscore the importance of gonadal hormones in mediating sexually dimorphic behavior and gene expression within areas of the brain fundamental to anxiety and depressive disorders. / A Dissertation submitted to the Department of Biomedical Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Summer Semester, 2012. / June 4, 2012. / anxiety, depression, ERK2, hippocampus, medial prefrontal cortex, sex differences / Includes bibliographical references. / Mohamed Kabbaj, Professor Directing Dissertation; Elaine Hull, University Representative; James Olcese, Committee Member; Michael Overton, Committee Member.

Due to the nutritional and health benefits, the consumption of fish as an important dietary protein source is increasing. However, the benefits are not always received by everyone. Fish is one of the eight major allergenic foods or food types and its prevalence in the United States was determined to be 0.4% of food-hypersensitive patients among the adult population. Approximately 1.1 million Americans suffer from various symptoms of fish allergy, and this number is increasing. Food manufacturers have been mainly depending on good manufacture practice (GMP) and hazards analyze and critical control point (HACCP) plan to avoid the presence of undeclared allergens in their products. Immunoassays using antibodies are widely accepted by regulatory agencies as a rapid and sensitive method for screening and monitoring substances in food and agricultural products. However, the vast species genetic variation of fish results in failure of identifying a single protein as a marker for fish protein detection. The specific objectives are: 1) to examine the selectivity of each MAbs 8F5, 2G9 and 2F3 by using indirect non-competitive ELISA(iELISA) and their antigenic proteins using western blot; 2) to develop a user friendly dot blot using pooled antibody for the detection of the presence of fish protein from all common food fish species; 3) to study the effects of storage and processing on the immunoreactivity thus the detestability of fish proteins using iELISA and dot blot. Three anti-fin fish MAbs (8F5, 2G9 and 2F3) were previously developed in our laboratory against cooked crude fish protein extract from red snapper, yellow fin tuna and swordfish, respectively. None of them was able to show the ideal pattern of recognition of the target fish species without cross reactivity or cross react with all fish species tested. But the cross reactivity pattern of each MAb complements to each other which may be utilized to recognize of a complete spectrum of most commercially important fish species. The overall goal of this research is to develop rapid user-friendly immunochemical detection methods using pooled MAb 8F5, MAb 2G9, and MAb 2F3 for the detection of fish proteins. The expected outcomes including: 1) Pooling of these three antibodies covered the immunoreactivity of commercially important fin fish and no cross reactivity with non-fish protein; the antigenic proteins of each MAb will also be identified; 2) pooling of MAbs would be effective for the detection of commercially important fish species using iELISA and dot blot assay; 3) iELISA and dot blot assay are effective and valid for fish detection under various storage and processing conditions. The pooled MAbs can be used as a standard reagent for detection of common fish species. Furthermore, a consumer-friendly immunoassay for the detection of the presence of fish could decrease the time of training experimental personnel. The research could provide a user-friendly tool not only to the regulatory agency for ensuring the enforcement of food laws but also the fish sensitive patients for risk reduction. / A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. / Spring Semester, 2013. / November 20, 2012. / Detection, Dot blot, ELISA, Fish, Pooled MAbs / Includes bibliographical references. / Yun-Hwa Peggy Hsieh, Professor Directing Thesis; Shridhar K. Sathe, Committee Member; Mary Ann Moore, Committee Member.

Purpose: The purpose of the present study is to evaluate the persistent effects of a 6-month intervention of resistance training on exercise behavior, physical activity levels, body composition, strength, and quality of life in female breast cancer survivors 18 months after completion of the resistance-training intervention. Methods: Twenty two out of the 27 female breast cancer survivors (63.5 ± 6.5 yrs old) agreed to participate from the original 6-month study. Height, weight and waist and hip circumferences were measured. Bone mineral density (BMD) and body composition were measured by Dual Energy X-ray Absorptiometry (DXA). Muscular strength including chest press one repetition maximal (1RM), leg extension 1RM and handgrip strength were measured. Resistance training status during the follow-up period was measured by the Resistance Training Status Questionnaire. Quality of life (QOL) scores including Physical Function, Mental QOL, and Physical QOL were measured by the Short Form 36 (SF-36) questionnaire. Data were analyzed with a Two-Way Repeated Measures Analysis of Variance. Significance was accepted at p<0.05. Results: Participants were grouped into compliers (n=10; trained for at least 6 months after completion of intervention) and non-compliers (n=12; those who did not continuing training). The women who fell into the complier group were women who were further out from the diagnosis of cancer (compliers: 116.1 ± 86.2 months vs non-compliers: 59.5 ± 35.9 months), had a lower stage of breast cancer diagnosis (compliers: 1.2 ± 0.6 vs non-compliers: 2.0 ± 0.6), and were more physically active (compliers: 7482 ± 2253 vs non-compliers: 4791 ± 2136 steps/day). Although there were no significant group by time interactions there were some time effects for the different variables. In both groups follow-up chest press and leg extension 1RM measures were significantly higher than baseline measurements (before original intervention), yet significantly lower than post measurements (after 6 month intervention). There was no difference in strength at follow-up between the two groups. There were significant time effects for lean mass (37.5 ± 5.1to 39.0 ± 5.8 kgs) and gynoid fat % (49.2 ± 2.7 to 46.1 ± 3.7 %) with the compliers having significant changes from baseline to follow-up with non-significant changes in the non-compliers. Although there were no group by time effects there were differences between the groups at follow-up Compliers had significantly less android fat (compliers: 39.4 ± 9.0 vs. non-compliers: 47.9 ± 8.3%), lower android/gynoid ratio (compliers: 0.85 ± 0.16 vs. non-compliers: 1.03 ± 0.14), lower waist girth (compliers: 82.5 ± 9.6 vs. non-compliers: 91.3 ± 13.0 cm) and higher lean body than non-compliers. BMD was not maintained for either group over the 18 months. BMD losses ranged from 1.1% to 4.1% at femur and forearm sites. Both compliers and non-compliers maintained quality of life through training study and follow-up time point. Conclusion: Continued resistance training and physical activity for at least six months following a structured intervention of resistance training helped breast cancer survivors improve some components of fat mass and lean mass. However, BMD was not maintained. Active life styles, time since diagnosis, and fat distribution may be indicators of exercise adherence rates. More research is needed to find interventions to help maintain or slow the loss of BMD in breast cancer survivors. / A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. / Summer Semester, 2013. / June 17, 2013. / breast cancer survivors, follow-up, resistance training / Includes bibliographical references. / Lynn Panton, Professor Directing Dissertation; Jeong-Su Kim, Committee Member; Robert Contreras, Committee Member.

Purpose: This study examined dietary and anthropometric components of diabetic patients with or without diabetic foot ulcers (DFU) and the association between advanced glycation end-products (AGE) and serum markers of oxidative stress and inflammation in diabetic patients with or without DFU. Methods: Eighty-two adult participants were recruited and assigned to one of three groups: 1) non-diabetic control; 2) diabetic participants without foot ulcers (DM); and 3) diabetic participants with foot ulcers (DFU). Twenty-four hour food recalls, pertinent history and blood samples were collected from each participant at the time of recruitment. Dietary intake was evaluated with Food Processor. Dietary and serum AGE as well as serum tumor necrosis factor-á (TNF-á), C-reactive protein (CRP), and thiobarbituric acid reactive substances (TBARS) were analyzed. Results: DFU subjects in this study were mostly overweight or obese. DFU had inadequate intakes in protein, fiber, vitamin B1, B2, B3, B6, C, D, and E; calcium, magnesium, phosphorus, potassium, selenium, and zinc. They had excessive intakes in saturated fat, trans fat, and sodium. The diabetic participants had significantly higher levels of dietary AGE per unit of energy, serum TNF-á, and TBARS compared to non-diabetic controls. BMI, body weight and TBARS were significantly higher in DFU than DM. DFU had significantly higher AGE per unit protein compared with DM and control. Serum AGE and TBARS were significantly correlated with dietary AGE. Serum TBARS predicted the duration of DFU (R2 = 0.52). Conclusions: Malnutrition is very common in the DM and DFU subjects. Vitamin supplementation may be beneficial in prevention and management of DM as well as DFU. Individuals with DFU had the highest levels of both dietary and serum AGE. Because dietary AGE causes a rise in serum AGE concentration, it is important to reduce the intake of foods containing AGE by promoting appropriate dietary choices in this population. / A Dissertation submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Spring Semester, 2013. / March 4, 2013. / diabetes melittus, oxidative stress / Includes bibliographical references. / Maria T. Spicer, Professor Co-Directing Dissertation; Jasminka Ilich-Ernst, Professor Co-Directing Dissertation; Cathy W. Levenson, University Representative; Jeong-Su Kim, Committee Member.

Ovariectomized (ovx) rats gain excess body weight (BW) even when they are pair-fed to sham animals. This higher BW, contrary to common belief, does not result in higher bone mineral density. The purpose of the present study was to evaluate the potential mechanisms underlying these paradoxical observations. We evaluated the effects of ovariectomy and 17β-estradiol (E2) using two age groups female Sprague-Dawley (SD) rats: 5- and 10-month old rats. Thirty-six female SD rats, 5- and 10-months old were acclimatized for 5 days. Animals in each category were divided into three groups: sham, ovx, and ovx+E2. There were 18 rats per age category with sample size of six rats per treatment group. Five- and ten-month old rats were chosen to model young and middle age women, respectively. Immediately after surgery, rats in all groups received a semi-purified control diet (AIN-93M). Animals in the ovx+E2 group were injected with E2 (10μ/kg BW subcutaneously, twice per week). Ovx control rats received solvent vehicle (sesame oil; subcutaneously, twice per week). Rats in ovx groups were pair-fed to the mean food intake of the sham group and their food intake was adjusted every three days. Rats' voluntary running activity was measured by activity wheel and distance counter in individual cages for 24 hours between one to two weeks before sacrifice. To confirm bone loss due to ovariectomy, whole body bone mineral density (BMD) was measured by dual energy x-ray absorptiometry (iDXA) at baseline, mid-point and at the end of study period. The losses of whole body BMD in five- and ten-month old ovx rats occurred after three and half- and five-months from the time of the removal of ovaries, respectively. In spite of pair feeding the ovx rats, the final body weights of ovx rats in both age categories were significantly (P<0.05) higher than those of sham-operated rats. Estrogen completely prevented the ovariectomy-induced weight gains in both age categories. Ovariectomy either significantly decreased (10-month old) or tended (P<0.1) to decrease (5-month old) voluntary wheel running activity and in comparison with sham animals. Estrogen treated rats voluntary wheel running activity was able to prevent this ovariectomy-induced decline in physical activity. Mean fat mass of ovx rats in comparison with sham animals was significantly higher in both age groups. Rats that received E2 did not experience gain in fat mass. The mean tibial and vertebral BMD values of ovx rats were significantly lower in comparison with sham rats in both age categories. Microstructural properties of tibiae and vertebrae, including bone volume/over total volume, connectivity density, structure model index were all unfavorably altered by ovariectomy in both age categories. In younger rats, E2 administration was able to prevent ovariectomy-induced alterations in lumbar vertebrae microstructural parameters such as bone volume/total volume (BV/TV), structure model index (SMI) and trabecular thickness (Tb. Th.). However, in this age group E2 administration was only able to prevent ovx-induced increase in tibial trabecular separation (Tb. Sp.). These findings suggest that E2 is more effective in preventing microstructural properties of trabecular rich bone such as vertebral bones. In older rats, in addition to the loss of BV/TV, ovx also caused significant decreases in connectivity density (Conn. D.) and trabecular number (Tb. N.) of both tibial and lumbar vertebral bones while increasing SMI and Tb. Sp. in the same bones. E2 administration was able to prevent changes in Conn. D. and SMI only in lumbar bones. E2 was also able to prevent the ovx-induced deleterious effects on tibial Tb.N. and Tb.Sp. by maintaining these values to those of sham levels. Overall, these observations suggest that while E2 is capable of preventing some structural properties as a result of ovx, it is not able to replace ovarian hormones as a whole. Ovariectomy significantly increased serum levels of both biomarker of bone resorption, C-telopeptides of type I collagen, and bone formation, alkaline phosphatase levels. These findings are in agreement with earlier reports that indicate ovariectomy increases the rate of bone turnover with resorption exceeding that of formation. Our findings do not suggest that the bone loss due to ovx is through inflammatory processes as the serum levels of tumor necrosis factor alpha (TNF-α) remained below the detectable level in ovx rats in both age categories. In spite of ovx-induced significant increases in systemic levels of insulin-like growth factor-I (IGF-I) in both age categories, its localized mRNA expressions were either significantly or tended (P < 0.1) to be reduced in gastrocnemius (GAS) and soleus (SOL) muscles of 10-month old rats. These local reductions in IGF-I mRNA expressions were in parallel with reductions in normalized to total body weight muscle mass, signifying the importance of IGF-I at the tissue level. Similar trends (P < 0.1) were also observed in 5-month old rats, albeit not significantly. Our findings for the first time, to our knowledge, suggest that elevated systemic IGF-I may down-regulate its synthesis in GAS and SOL of ovx rats. Overall, the findings of this study indicate that ovariectomy causes significant losses of BMD, microstructural properties and muscle mass. However, body fat increases as such that the weight of ovx animals exceeds those of sham and E2 treated animals, which indicates that ovarian hormone deficiency causes body weight gain mainly due to a gain in fat mass. Additionally, our data suggest that the detrimental effects of ovarian hormone deficiency are exacerbated as a result of aging. Our observations, especially in older rats, suggest that estrogen deficiency alters body composition which resembles that of osteosarcopenic obesity. / A Dissertation submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Spring Semester, 2013. / November 26, 2012. / Body Composition, Estrogen, Menopause, Obesity, Osteosarcopeni Obesity, Sarcopenia / Includes bibliographical references. / Bahram H. Arjmandi, Professor Directing Dissertation; P. Bryant Chase, University Representative; Wayne Denton, Committee Member; Arturo Figueroa, Committee Member; Jeong-Su Kim, Committee Member.

Although the use of hemoglobin-containing food additives is increasing, there is virtually no method available for monitoring their presence in food and dietary products. We have previously developed a competitive ELISA (cELISA) for the detection of bovine blood in food using a monoclonal antibody (MAb), Bb1H9, which recognizes a 12kDa antigenic protein. Because 12kDa approximates that of a monomer of hemoglobin (14.4kDa) and further studies have shown the 12kDa protein to be present in the red cells of bovine blood, we hypothesized that this protein is a subunit of hemoglobin. This study sought to verify this hypothesis and hence investigate the usefulness of assays based on MAb Bb1H9 in monitoring the presence of bovine hemoglobin-containing food additives. Extracts obtained from raw and heat-treated bovine blood fractions and products comprising of bovine hemoglobin-containing ingredients were analyzed with indirect ELISA (iELISA), cELISA and western blot. Target proteins resolved by two-dimensional electrophoresis were subjected to N-terminal sequencing. Results indicated that the 12kDa protein is a monomer of the tetrameric hemoglobin molecule (64.5kDa) and that MAb Bb1H9 was able to bind both in the presence and in the absence of heme. MAb Bb1H9 was also found to react with a linear (ERMFLSF) epitope on the alpha chain and possibly, a conformational epitope on the beta subunit of hemoglobin. MAb Bb1H9 in cELISA format proved useful against all products tested except for the bovine hemoglobin hydrolysate (RIF) and sodium erythorbate (DAU) containing products. MAb Bb1H9 in western blot format, however, proved useful for the detection of the product, DAU. MAb Bb1H9 also proved useful in the detection of bovine hemoglobin hydrolysates that have been hydrolyzed with pepsin for up to 6h. MAb Bb1H9 is expected to become a valuable regulatory tool for labeling law enforcement to protect the interest of individuals (e.g. Jews, Muslims, and vegans) that avoid consuming blood-derived products for religious or ethical reasons; and to deter rogue manufacturers who may be tempted to use such blood-derived proteins fraudulently to increase the apparent meat content. / A Dissertation submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Fall Semester, 2011. / October 7, 2011. / Bovine, competitive ELISA, Hemoglobin, Monoclonal antibody / Includes bibliographical references. / Yun-Hwa Peggy Hsieh, Professor Directing Dissertation; Kenneth H. Roux, University Representative; Shridhar K. Sathe, Committee Member; Jeong-Su Kim, Committee Member.

The phytochemical composition and antioxidant capacity of blueberries support their role in reducing inflammation and oxidative stress, as well as other processes that contribute to the development of cancer. Hence, it is suggested that the consumption of blueberries on a regular basis reduces the risk of developing cancer. Though it is difficult to prove the cancer-preventive effects of any agent including foods and dietary supplements, epidemiological findings indicate diets rich in fruits, especially berries, are associated with a reduced risk of various types of cancer. In fact, there is a wealth of preclinical research findings suggesting a role for blueberries in cancer prevention. Although there are numerous risk factors predisposing individuals to cancer development, epidemiological evidence suggests a positive association between hypertension (HTN) and cancer mortality, particularly renal cancer. Though there are a few clinical studies that have examined the antihypertensive effects of blueberries, to date there are no studies investigating these effects in postmenopausal women with pre- and stage 1-HTN. The hypothesis for the present study was that the daily consumption of blueberries would reduce blood pressure (BP), inflammation, oxidative stress, and DNA damage and increase the production of cellular antioxidants in postmenopausal women with pre- and stage 1-HTN. If the consumption of blueberries positively affects some of these variables, this would indirectly demonstrate the anti-cancer properties of blueberries. We carried out an 8-week, randomized, double-blind clinical study in which a total of 40 women (n = 20 subjects per treatment group), 1 to 10 years postmenopausal, between the ages of 45 and 65 years with pre- (130/85 to 139/89 mm Hg) or stage 1-HTN (140/90 to 159/99 mm Hg), and otherwise healthy were eligible to participate. Qualified subjects were assigned to one of the two treatment groups: 22 g freeze-dried blueberry powder per day or 22 g nutrient-matched placebo per day for 8 weeks. To test our hypothesis, the following specific aims were used: 1) to investigate the extent to which the daily consumption of blueberries would reduce BP; 2) to determine the degree to which the daily consumption of blueberries would reduce serum markers of inflammation, including tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) as well as increase the anti-inflammatory marker, adiponectin. Additionally, this aim assessed the ratio between the proinflammatory marker leptin and adiponectin; and 3) to examine whether the daily consumption of blueberries would improve cellular antioxidant defense mechanisms by assessing blood levels of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR). Oxidative stress markers including nitric oxide (NO), 8-isoprostane, malondialdehyde (MDA), oxidized low-density lipoprotein (oxLDL), and the DNA damage marker, 8-hydroxy-2'-deoxyguanosine (8-OHdG) were also measured. Our results indicate a significant reduction in systolic and diastolic BP (- 5.1%, P < 0.05 and - 6.25%, P < 0.01, respectively) at 8 weeks whereas there were no significant decreases in BP in the control group. NO levels were significantly (P < 0.01) increased in the blueberry group at 8 weeks compared to baseline whereas there were no changes in the control group. While there were no significant within-group changes in the mean levels of 8-OHdG over time, there was a significant (P < 0.05) difference between groups at 4 weeks of treatment such that levels were lower in the blueberry group and there was a significant (P = 0.04) group by time interaction. The remaining biochemical markers measured in this study were not affected by the blueberry treatment. Overall, the findings of this study suggest that blueberry is capable to reduce both systolic and diastolic BP, most likely due to its ability to upregulate the synthesis of NO. Additionally, the consumption of blueberries for 4 weeks decreases DNA damage although not significant and there is an increase in DNA damage between 4 and 8 weeks. The most plausible explanation for this occurrence is that with the increase in NO between 4 and 8 weeks there may have been an increase in the production of peroxynitrate which is known to cause DNA damage, or there could have been an inhibition of DNA repair enzymes. These findings suggest that short-term consumption (4 weeks) of blueberries indirectly exerts cancer-protective properties while longer-term (8 weeks) consumption exerts antihypertensive effects. Future in vivo studies are needed to elucidate the mechanisms of these findings, as well as to determine the longer-term outcomes in terms of cancer and hypertension-related diseases. / A Dissertation submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Fall Semester, 2013. / October 28, 2013. / Antioxidants, DNA damage, Functional Foods, Hypertension, Nutrition, Phytochemicals / Includes bibliographical references. / Bahram H. Arjmandi, Professor Directing Dissertation; Cathy W. Levenson, Outside Committee Member; Maria T. Spicer, Committee Member; Carla M. Prado, Committee Member.

BACKGROUND: Epidemiological studies have shown that cardiovascular events are more prevalent in the winter than in other season. Although the exact mechanisms are not completely understood, previous findings indicate that acute cold exposure increases sympathetic activity, arterial stiffness (pulse wave velocity [PWV]), and blood pressure (BP) as well as impairs endothelial function. Moreover, obesity is characterized by increased sympathetic activity and BP, which may further represent a potential mechanism responsible for cardiovascular events. There is increasing evidence that recovery from an acute bout of exercise is a vulnerable phase for cardiac events. Therefore, the elucidation of cardiovascular responses to post-exercise recovery concurrent with cold exposure in overweight/obese individuals is of clinical importance. Submaximal isometric-handgrip (IHG) exercise has been previously used as a means for evaluating the role of the exercise pressor reflex (muscle metaboreflex) on the cardiovascular system and cardiac autonomic function. The exercise pressor reflex can be evaluated using the technique of post-exercise muscle ischemia (PEMI), which by trapping metabolites in a previously exercised muscle, sustains the vascular sympathetic stimulation evoked during exercise in a controlled fashion. Recently, oral supplementation with the amino acid L-citrulline (L-cit) has been proposed as a possible adjunct treatment for hypertension and arterial stiffness. L-cit is known to enhance the bioavailability of L-arginine, the endothelial substrate for the potent vasodilator nitric oxide (NO). Our group demonstrated that L-cit supplementation effectively attenuates the hemodynamic responses to cold pressor test (CPT); however, the potential cardio-protective effects of L-cit supplementation to reduce cold-induced hypertension (CIH) during exercise pressor reflex activation have yet to be evaluated. PURPOSE: The aim of this study was twofold. 1) To evaluate the acute effects of the exercise pressor reflex imposed by PEMI and PEMI concurrent with CPT (PEMI+CPT) on hemodynamics, arterial stiffness, and cardiac autonomic modulation in healthy overweight/obese men, and 2) to examine the effects of a 14-day course of L-cit supplementation on hemodynamics, arterial stiffness, and cardiac autonomic modulation during IHG exercise, PEMI, and PEMI+CPT in healthy overweight/obese men. METHODS: Sixteen healthy young (24 ± 1.5 y) overweight/obese men were randomly assigned to receive either placebo (maltodextrin) or L-cit (6 g/day) for 14 days in a cross-over fashion. Following 5 min of resting measurements, the participants were asked to perform two trials in a randomized order. In one trial, the participants perform a 2-min IHG exercise (30% of maximal voluntary contraction) followed by a 3-min PEMI. Following a 15-min recovery period, the participants were asked to perform a 2-min IHG exercise followed by a 3-min PEMI concurrent with CPT. Brachial systolic BP (bSBP), brachial diastolic BP (bDBP), aortic systolic BP (aSBP), aortic diastolic BP (aDBP), augmentation index (AIx), first (P1) and second systolic peak (P2, wave reflection magnitude), brachial-ankle PWV (baPWV, arterial stiffness), heart rate (HR), and HR variability (HRV, cardiac autonomic modulation) were evaluated at rest, during IHG, PEMI, and PEMI+CPT. All measurements were re-evaluated after 14 days. RESULTS: Height, weight, body mass index, and waist circumference were 1.72 ± 0.01 m, 86.8 ± 3.7 kg, 29.3 ± 1.1 kg/m2, and 98 ± 3 cm, respectively. At baseline, all hemodynamic parameters increased (P<0.01) during IHG exercise and remained elevated during PEMI and PEMI+CPT. BSBP, bDBP, aSBP, aDBP, P1, and P2 were higher (P<0.01) during PEMI+CPT compared to PEMI. Sympathetic activity increased similarly during IHG and PEMI with a further increase during PEMI+CPT. During PEMI+CPT, HR and low frequency components of the HRV (LnLF and nLF, markers of sympathetic activity) were higher (P<0.05) whereas the high frequency (LnHF), a marker of parasympathetic activity, was decreased (P<0.01) compared to PEMI. There were no significant changes in all hemodynamic parameters at rest after the treatments. During IHG, L-cit significantly decreased bDBP (-10 ± 3 mmHg; P<0.01), aSBP (-8 ± 3 mmHg; P<0.05), AIx (-14.2 ± 3.1 %; P<0.05), and P2 (-8 ± 3 mmHg; P<0.01) compared to no changed after placebo. During PEMI, L-cit significantly decreased bDBP (-10 ± 3 mmHg; P<0.05), aDBP (-8 ± 2 mmHg; P<0.01), and AIx (-8.6 ± 2.0 %; P<0.01) compared to no changes after placebo. During PEMI+CPT, L-cit significantly decreased bSBP (-11 ± 3 mmHg; P<0.01), bDBP (-11 ± 2 mmHg; P<0.01), aSBP (-13 ± 3 mmHg; P<0.01), aDBP (-10 ± 2 mmHg; P<0.01), AIx (-13.8 ± 2.2 %; P<0.01), P1 (-7 ± 2 mmHg; P<0.01), P2 (-14 ± 3 mmHg; P<0.01) and baPWV (-0.9 ± 0.4 m/s; P<0.05). There were no significant changes in all HRV parameters after the treatments. CONCLUSIONS: The present study demonstrates that post-exercise metaboreflex activation concurrent with cold exposure imposes an additional cardiovascular stress due to increased sympathetic activity. In addition, we showed that L-cit supplementation attenuates the exercise pressor responses imposed by IHG exercise and PEMI. Furthermore, we showed that L-cit supplementation is an effective means to decrease the CIH responses during post-exercise metaboreflex activation. In conclusion, L-cit supplementation may be a potential adjunct treatment to reduce the CIH responses and ultimately provide cardio-protective effect to those who perform exercise in low environmental temperatures. / A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. / Spring Semester, 2014. / March 28, 2014. / Includes bibliographical references. / Arturo Figueroa, Professor Directing Thesis; Michael Ormsbee, Committee Member; Gershon Tenenbaum, Committee Member.