Doctor of Philosophy(PhD) / Several studies have documented the direct effect of macronutrient intake on sleep. A general picture that has emerged indicates that a low carbohydrate diet with a total energy between 13-47% and high fat content with a total energy between 47-77% shows increases in slow wave sleep and may decrease rapid-eye movement sleep. However, previous studies investigating the association between carbohydrate meals and sleep have not explored the effects of the glycemic index (GI) of carbohydrate on sleep. This thesis investigated the affect of GI on the sleep pattern. In a cross-over, repeated measures design, we explored both the effect of GI and the timing of these meals on sleep in good sleepers. The effects of high and low GI carbohydrate-based meals given 4 h before the subjects’ usual bedtime on their sleep quality were examined in Chapter 3. Also evaluated was the effect of high GI meal timing (4 h vs. 1 h) on sleep. Twelve healthy men (18-35y, BMI 18.5-25 kgm-2) were administered a standard, isocaloric meal of low GI = 50 or high GI=109 in a cross-over and counter balanced manner, 4 h before their usual bedtime. On another occasion, the high GI meal was given 1 h before bedtime. Following the high or the low GI meal, participants underwent a familiarization sleep night followed by three polysomnographic test nights. The subjects’ blood and urine were collected for glucose and 6-sulfatoxymelatonin analysis respectively. Significant differences were found between the area under the curve (AUC) for blood glucose responses following the high GI meal compared to the responses for the low GI meal. It was shown that a carbohydrate-based high GI meal resulted in a significant shortening of sleep onset latency (SOL) in normal sleepers compared to a low GI meal (P = 0.009), and was most effective when consumed 4 h before bedtime (P = 0.01). There were no significant changes in other sleep indices. The Atkins’ Diet is a popular dietary therapy that promotes weight loss. This restricted carbohydrate diet with high fat and high protein content has not been evaluated for its effects on sleep, or systematically documented for its effects on mood, fatigue or sleepiness. The short term effect of the Atkins’ diet over 48 h on the sleep quality of healthy, non-obese males to a Control mixed diet was compared in Chapter 4. This study employed a repeated measure design where fourteen healthy, non-obese, good sleepers were given isocaloric diets and matching evening test meals (4 h before usual bedtime), which were either mixed (15% protein, 25% fat, 60% carbohydrate) or Atkins’ (38% protein, 61% fat, <1% carbohydrate). After a familiarization night with polysomnography, further polysomnographic testing was then performed on the Control night, 4 h after the first Atkins’ test meal (Atkins Acute) and 48 h (Atkins Ketosis) following commencement of the Atkins’ diet. Objective sleep was recorded using Compumedics S-series Sleep system; Compumedics Ltd, Melbourne, Australia. Urine ketone level was monitored before the evening test meals and at bedtime on the Control night, during the Atkins Acute and Ketosis phase. Blood glucose level was measured before the evening test meal until 120 min following the meal. Significant differences were found for the AUC for the blood glucose between the Control night and the Atkins Acute and Atkins Ketosis phase (P < 0.001). Participants developed mild hypoglycemia and ketosis 48 h following the Atkins’ diet. A significant reduction in the proportion of rapid eye movement (%REM) sleep to total sleep time (TST) was observed following the Atkins’ Acute and Atkins’ Ketosis phase compared to the Control (P = 0.006 and 0.05 respectively). The percentage of slow wave sleep (%SWS) to TST significantly increased for both the Atkins’ Acute and Ketosis phase compared to the Control meal (P = 0.02 for both phases). The sleep changes may be linked to the energy metabolism of fat of the Atkins’ diet. The effects of the Atkins’ diet compared to a Control mixed diet on sleepiness, mood, fatigue and dream recall were also investigated (Chapter 5). Participants’ overall daytime mood, fatigue intensity, sleepiness and other symptoms were assessed using a visual analogue scale before the evening test meals. The number of subjects with dream recalls was recorded on awakening after each polysomnographic night. The daytime symptoms of fatigue, sleepiness and depressed mood were significantly increased following the Atkins’ diet compared to the Control diet. A greater proportion of subjects reported dreams 48 h after the Atkins’ diet compared to either the Atkins’ Acute phase or the Control condition. Our findings suggest that mild hypoglycemia resulting from the diet may mediate the subjective responses of daytime sleepiness, depressed mood and intense fatigue. The increased proportion of subjects with dream recall may be related to an increased transient arousals from sleep during which dreams are usually consolidated into memory. The finding that “high-glycemic-index carbohydrate meals shorten sleep onset” may be relevant to persons with sleep disturbance. These meals may facilitate sleep transition for those with sleep initiation problems. The effect of the Atkins’ diet in SWS promotion and increasing feelings of fatigue and suppressing mood in the short-term may be relevant for patients with sleep apnoea (obesity), who experience low proportion of SWS and significant somnolence. Further studies to explore these effects on a longer term in this group would be worthwhile.

Doctor of Philosophy(PhD) / Several studies have documented the direct effect of macronutrient intake on sleep. A general picture that has emerged indicates that a low carbohydrate diet with a total energy between 13-47% and high fat content with a total energy between 47-77% shows increases in slow wave sleep and may decrease rapid-eye movement sleep. However, previous studies investigating the association between carbohydrate meals and sleep have not explored the effects of the glycemic index (GI) of carbohydrate on sleep. This thesis investigated the affect of GI on the sleep pattern. In a cross-over, repeated measures design, we explored both the effect of GI and the timing of these meals on sleep in good sleepers. The effects of high and low GI carbohydrate-based meals given 4 h before the subjects’ usual bedtime on their sleep quality were examined in Chapter 3. Also evaluated was the effect of high GI meal timing (4 h vs. 1 h) on sleep. Twelve healthy men (18-35y, BMI 18.5-25 kgm-2) were administered a standard, isocaloric meal of low GI = 50 or high GI=109 in a cross-over and counter balanced manner, 4 h before their usual bedtime. On another occasion, the high GI meal was given 1 h before bedtime. Following the high or the low GI meal, participants underwent a familiarization sleep night followed by three polysomnographic test nights. The subjects’ blood and urine were collected for glucose and 6-sulfatoxymelatonin analysis respectively. Significant differences were found between the area under the curve (AUC) for blood glucose responses following the high GI meal compared to the responses for the low GI meal. It was shown that a carbohydrate-based high GI meal resulted in a significant shortening of sleep onset latency (SOL) in normal sleepers compared to a low GI meal (P = 0.009), and was most effective when consumed 4 h before bedtime (P = 0.01). There were no significant changes in other sleep indices. The Atkins’ Diet is a popular dietary therapy that promotes weight loss. This restricted carbohydrate diet with high fat and high protein content has not been evaluated for its effects on sleep, or systematically documented for its effects on mood, fatigue or sleepiness. The short term effect of the Atkins’ diet over 48 h on the sleep quality of healthy, non-obese males to a Control mixed diet was compared in Chapter 4. This study employed a repeated measure design where fourteen healthy, non-obese, good sleepers were given isocaloric diets and matching evening test meals (4 h before usual bedtime), which were either mixed (15% protein, 25% fat, 60% carbohydrate) or Atkins’ (38% protein, 61% fat, <1% carbohydrate). After a familiarization night with polysomnography, further polysomnographic testing was then performed on the Control night, 4 h after the first Atkins’ test meal (Atkins Acute) and 48 h (Atkins Ketosis) following commencement of the Atkins’ diet. Objective sleep was recorded using Compumedics S-series Sleep system; Compumedics Ltd, Melbourne, Australia. Urine ketone level was monitored before the evening test meals and at bedtime on the Control night, during the Atkins Acute and Ketosis phase. Blood glucose level was measured before the evening test meal until 120 min following the meal. Significant differences were found for the AUC for the blood glucose between the Control night and the Atkins Acute and Atkins Ketosis phase (P < 0.001). Participants developed mild hypoglycemia and ketosis 48 h following the Atkins’ diet. A significant reduction in the proportion of rapid eye movement (%REM) sleep to total sleep time (TST) was observed following the Atkins’ Acute and Atkins’ Ketosis phase compared to the Control (P = 0.006 and 0.05 respectively). The percentage of slow wave sleep (%SWS) to TST significantly increased for both the Atkins’ Acute and Ketosis phase compared to the Control meal (P = 0.02 for both phases). The sleep changes may be linked to the energy metabolism of fat of the Atkins’ diet. The effects of the Atkins’ diet compared to a Control mixed diet on sleepiness, mood, fatigue and dream recall were also investigated (Chapter 5). Participants’ overall daytime mood, fatigue intensity, sleepiness and other symptoms were assessed using a visual analogue scale before the evening test meals. The number of subjects with dream recalls was recorded on awakening after each polysomnographic night. The daytime symptoms of fatigue, sleepiness and depressed mood were significantly increased following the Atkins’ diet compared to the Control diet. A greater proportion of subjects reported dreams 48 h after the Atkins’ diet compared to either the Atkins’ Acute phase or the Control condition. Our findings suggest that mild hypoglycemia resulting from the diet may mediate the subjective responses of daytime sleepiness, depressed mood and intense fatigue. The increased proportion of subjects with dream recall may be related to an increased transient arousals from sleep during which dreams are usually consolidated into memory. The finding that “high-glycemic-index carbohydrate meals shorten sleep onset” may be relevant to persons with sleep disturbance. These meals may facilitate sleep transition for those with sleep initiation problems. The effect of the Atkins’ diet in SWS promotion and increasing feelings of fatigue and suppressing mood in the short-term may be relevant for patients with sleep apnoea (obesity), who experience low proportion of SWS and significant somnolence. Further studies to explore these effects on a longer term in this group would be worthwhile.

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RATIONALE: Tuberculosis and HIV infection together form a highly mortal combination. Even after the advent of highly active antiretroviral therapy (HAART) medications, management for Tuberculosis and HIV/AIDS still remains a challenge. Poor outcomes (in both morbidity and mortality) are still being witnessed throughout the world, and especially in the poorly developed countries that bear the bulk of the burden of the cases. It is assumed that one of the major contributors to the poor outcomes is poor nutritional status resulting from the disease process itself, poverty and toxicity from medications being used to treat these diseases that substantially reduce appetite. An assessment of the role that nutritional status has on change in CD4 as a surrogate marker of disease progression is therefore of importance. OBJECTIVES: To evaluate the role that nutritional supplementation has on change in CD4 in TB patients co-infected with HIV who are receiving standard care of treatment. METHODS: Data from a randomized controlled trial of a Protein Calorie Supplement (PCS) were used. To assess the effect of randomization to a nutritional supplement, baseline characteristics were compared among the intervention and the control groups and confounder variables, such as age, BMI, baseline CD4, socioeconomic status, previous exposure to TB and compliance with HAART medication were analyzed and adjusted for in a model using multivariate linear regression. RESULTS: 151 HIV-infected women with TB disease were enrolled; 72 received PCS while 79 did not. We found that the PCS intervention had no significant effect on change in CD4 between baseline and 8 months. Average change in CD4 count was similar for intervention and control groups (204 vs. 207 units). This similarity persisted after adjusting for baseline BMI and previous TB disease. CONCLUSION: Randomization (i.e. nutritional supplement) did not have a significant effect on change in CD4 count among study participants. However, an effect could have been masked by high compliance with ART.

Master of Science / Department of Agricultural Economics / Vincent Amanor-Boadu / Metabolic syndrome (MetS) continues to be a public health concern in the United States. The current prevalence rate is about 34% among American adults. One of the recommended line of treatment for the components of MetS is dietary behavior change. Although, many dietary recommendations guidelines are published to aid in better dietary choices, little is known about how effectively they alter dietary choices. Thus, the overall objective of this study was to examine the extent to which knowledge about the presence of metabolic syndrome components influenced macronutrient intake. Data from 2013-2014 National Health and Nutrition Examination Survey (NHANES) were used for the study. The variables used were taken from modules of the NHANES dataset: demographic, dietary (day 1 and 2 recall), questionnaire (blood Pressure & Cholesterol, medical condition, diabetes and weight history), examination (blood pressure and body measures) and laboratory (cholesterol – high density lipoprotein, and triglycerides and plasma fasting glucose). Daily macronutrients (calories, protein, carbohydrate, fat and total sugar) intake were regressed on knowledge of MetS components presence and demographic characteristics using Ordinary Least Square model. The results show that having information that one has diabetes was associated with a reduced intake of daily calories (160 kcal), carbohydrate (22.73 g) and total sugar (15.26 g). There was no significant association between protein and fat intakes and the knowledge of the presence of a metabolic syndrome component in the econometric model. Ageing was associated with increase in calorie (16 kcal/day), protein (0.502 g/day) and fat (0.66 g/day) intake. Males consumed higher amounts of all macronutrients than females. Higher education was associated with higher fat intake (5.09 g/day for High School and 4.54 g/day for college compared with those with less than high school education) but reduced sugar intake (8.86 g/day) for those with college education. It was found that 27.59% of individual’s who had diabetes did not know they had it, and about 41% of those who did know they were overweight had central obesity. The study concludes that compared to knowledge about high triglyceride levels, low high-density lipoprotein, diabetes, high blood pressure and overweight, knowledge about having diabetes seems to motivate people to change their dietary intake. This may be due to the immediate effect of diet on diabetic patients compared to the other MetS components. The result of this is that it may be appropriate to pursue drug therapy for addressing the other MetS components while diet change may be effective contributor to managing diabetes.

<p> Resistance exercise training results in skeletal muscle hypertrophy due to accumulated periods ofprotein accretion, the magnitude ofwhich may be influenced by post-exercise feeding. One study suggests that hypertrophy is maximized by immediate provision ofprotein (26); how the type ofprotein ingested in this period affects protein accretion is not known. PURPOSE: To investigate the effect of consuming different post-exercise beverages -milk (MLK), an isonitrogenous and isoenergetic soy drink (PEC), or an isoenergetic control (CON)-on body mass, fat-bone free mass (FBFM), muscle fibre area, and strength during a 12-week progressive whole body resistance training program, in untrained men. METHODS: 34 men were randomized to postexercise consumption ofMLK (n=12; fat free), PEC (n=11; soy) or CON (n=11; maltodextrin) using single-blinded allocation, with characteristics-22.5 ± 0.6 yr, 25.6 ± 0.7 kg/m2• Participants trained 5 d/wk using a whole body split resistance training program and consumed 500 ml oftheir assigned drink immediately and 1h post-exercise following every training bout. RESULTS: Total body mass increased following training (P<0.01) but there were no differences between groups (MLK = 3.2 ± 0.8 kg, PEC = 2.9± 1.4 kg, CON= 2.0 ± 0.8 kg). Similarly, FBFM also increased post-training (P<0.01), but was not different between groups (MLK = 3.3 ± 0.6 kg, PEC = 2.7 ± 0.6kg, CON= 2.2 ± 0.6 kg). Muscle type II fibre area increased post-training (P<0.05) but was not different between groups (MLK = 1004 ± 249 pm2, PEC = 650 ± 192 pm2, CON= 565 ± 120 pm2). Training resulted in increases in 1RM strength for 13 different exercises (P<0.01) with no difference between groups (range= 29-101%). CONCLUSION: Immediate post-exercise provision ofeither milk (MLK), or an isonitrogenous I isoenergetic soy drink (PEC), as well as an isoenergetic carbohydrate beverage (CON), resulted in similar increases in body mass, FBFM, type II muscle fibre area, and strength. This study demonstrates that intact dietary proteins from milk and soy are effective for promoting skeletal muscle hypertrophy. Moreover, when given equivalent energy immediately postexercise, proteins consumed as part of a normal diet, outside ofthe immediate postexercise period are sufficient for hypertrophic and strength gains. Finally, post-exercise consumption ofprotein is no more effective than carbohydrate for promoting skeletal muscle hypertrophy when adequate protein is consumed within a regular diet. </p> / Thesis / Master of Science (MSc)

Prolactin-releasing peptide (PrRP) is an endogenous hypothalamic neuropeptide that when exogenously injected increases food intake in chickens, but decreases it in rodents and goldfish. We designed three sets of experiments to elucidate the mechanisms of PrRP's orexigenic effect in chicks. In experiment one, food and water intake were evaluated in chicks after receiving intracerebroventricular (ICV) injection of the vehicle, 0.75, 3, 12, 47 or 188 pmol PrRP. The administration of 12 and 47 pmol PrRP increased food intake for up to 120 min after injection, and 188 pmol increased it for up to 180 min. The lowest effective dose was 3 pmol, which increased food intake for up to 60 min after injection. Water intake was not affected. To investigate the molecular mechanisms, c-Fos immunohistochemistry was performed and mRNA expression of some appetite-associated neurotransmitters was measured in chicks that received either vehicle or 188 pmol of PrRP. The rostral paraventricular nucleus (PVN) was activated which coincided with increased neuropeptide Y (NPY) mRNA expression in the whole hypothalamus. In experiment two, food and water intake were evaluated in chicks fed a high carbohydrate (HC), high fat (HF) or high protein (HP) diet after ICV injection of vehicle, 3 or 188 pmol PrRP. Chicks fed the HP diet increased food intake at a lower dose than chicks fed HF and HP diets after ICV PrRP injection. In addition, ICV injection of vehicle, 3 and 188 pmol PrRP were performed in chicks fed all three diets, and ICV PrRP injection induced preferential intake of the HP diet over HC and HF diets. The expression of some appetite-associated neuropeptides in the hypothalamus was also measured in chicks fed the HC, HF or HP diet after ICV injection of vehicle or 188 pmol PrRP. There was a diet effect on mRNA abundance of all appetite-associated genes measured (P < 0.05), with greater expression in chicks fed the HF or HP than HC diet. While neuropeptide Y (NPY) mRNA abundance was similar between vehicle and PrRP-injected chicks that consumed HP or HF diets, expression was greater (P < 0.05) in PrRP- than vehicle-injected chicks that consumed the HC. In experiment three, the orexigenic effect of PrRP was tested in chicks selected for low (LWS) and high (HWS) body weight after central administration of vehicle, 24, 94 and 375 pmol PrRP. The LWS chicks had a lower threshold and higher magnitude of food intake increase in response to PrRP injection. Results demonstrate that PrRP is a potent orexigenic factor in chickens and that effects are likely mediated through the hypothalamus. The orexigenic effect of PrRP was influenced by dietary macronutrient composition, and diet in turn influenced the food intake response to PrRP. These results may contribute to a novel understanding of appetite regulation. / Master of Science

The objective of this dissertation research was to investigate the effect of dietary macronutrient composition on neuropeptide Y (NPY)-mediated regulation of adipose tissue physiology in broiler chickens during the early post-hatch period. A high-carbohydrate (HC), high-fat (HF) or high-protein (HP) diet was fed to broiler chicks in all experiments and various facets of physiology were evaluated at day 4 post-hatch, including diet-, fasting-, and neuropeptide Y-induced effects on gene expression, cellular morphology, and lipid metabolism. Experiment 1 was designed to study the effects of diet on molecular changes in different adipose tissue depots (subcutaneous, clavicular and abdominal) after 3 hours of fasting and 1 hour of refeeding. Adipose tissue weights were decreased in chicks that consumed the HP diet, whereas adipocyte diameter was increased in response to the HF diet. There was greater expression of mRNAs encoding fatty acid binding protein 4 (FABP4) and monoglyceride lipase in chicks fed the HC and HF diets than the HP diet in all three adipose tissue depots. Fasting increased plasma non-esterified fatty acid concentrations in chicks fed the HC and HP diets. Results suggest that the heavier fat depots and larger adipocytes in chicks fed the HF diet are explained by greater rates of hypertrophy, whereas the HP diet led to a decrease in adipose tissue deposition, likely as a result of decreased rates of adipogenesis. Experiments 2 and 3 were designed to investigate how dietary macronutrient composition affects the effect of centrally or peripherally administered NPY, respectively, on lipid metabolism-associated factor mRNAs in adipose tissue. In experiment 2, vehicle or 0.2 nmol of NPY was injected intracerebroventricularly (ICV) and abdominal and subcutaneous fat samples were collected at 1 hour post-injection. In the subcutaneous fat, ICV NPY injection decreased peroxisome proliferator-activated receptor gamma (PPAR gamma) and sterol regulatory element-binding transcription factor 1 (SREBP1) mRNAs in chicks fed the HF diet, whereas there was an increase in SREBP1 expression in chicks fed the HF diet after NPY injection. Expression of PPAR gamma and FABP4 mRNAs increased in the abdominal fat of HF diet-fed chicks after NPY injection. Thus, HF diet consumption may have enhanced the sensitivity of chick adipose tissue to the effect of centrally-injected NPY on gene expression of adipogenesis-associated factors. In experiment 3, vehicle, 60, or 120 micrograms/kg BW of NPY was injected intraperitoneally (IP), and subcutaneous, clavicular, and abdominal fat was collected at 1 and 3 hours post-injection. Food intake and plasma NEFA concentrations were not different among chicks fed the HC, HF or HP diet after IP NPY injection, indicating that the effects of NPY on adipogenesis were independent of secondary effects due to altered energy intake. In response to the lower dose of NPY, the expression of NPY receptor sub-type 2 mRNA was increased at 1 hour post-injection in the subcutaneous fat of chicks fed the HP diet, whereas there was less 1-acylglycerol-3-phosphate O-acyltransferase 2 mRNA in the subcutaneous fat of chicks fed the HC diet. The higher dose of NPY was associated with greater AGPAT2 mRNA in the clavicular fat of chicks that consumed the HP diet and less CCAAT/enhancer-binding protein alpha in the abdominal fat of chicks that were provided the HF diet. However, there was also a decrease in the expression of some of these factors, although mechanisms are unclear. In conclusion, dietary macronutrient composition influenced the response of adipose tissue to the adipogenic effects of NPY and metabolic effects of short-term fasting and refeeding during the first week post-hatch. Collectively, this research may provide insights on understanding NPY's effects on the development of adipose tissue during the early life period and mechanisms underlying diet-dependent and depot-dependent differences in adipose tissue physiology across species. / Ph. D.

Understanding the central nervous systems role in appetite regulation is crucial to cure the obesity epidemic, which is more prevalent than any disease in the United States. Central appetite regulators, known as neuropeptides, are pivotal in understanding appetite regulation. Neuropeptide Y (NPY), a 36 amino acid peptide, plays a major role in regulating the hunger signals from the brain. In all vertebrates studied, it is a strong orexigenic neurotransmitter located throughout multiple nuclei of the hypothalamus. Peripheral hormones associated with hunger are able to activate NPY neurons in the arcuate nucleus, which leads to a cascade of events that activate orexigenic neurons throughout the hypothalamus. Although extensive research has gone into understanding the role of NPY in appetite regulation, the effects of macronutrient composition of diets on NPY function have not been elucidated in non-mammalian species. This research investigates how food intake is affected by dietary macronutrient composition in broiler type chickens that are fed three varying macronutrient diets: high carbohydrate (22% CP, 3000kcal/kg) a broiler starter diet, high fat (60% ME from lard), high protein 30%CP). All diets were formulated to be isocaloric. When chicks are fed the high fat diet central NPY administration has a greater effect on feed intake compared to both the basal and high protein diet. Regardless of what diet the chick is fed from hatch, if they are switched to one of the other two diets post central administration of NPY the high fat diet stimulated feed intake for the longest duration. Although, NPY had the strongest orexigenic effect on chicks fed the high fat diet, in a choice diet situation broiler chicks chose the high protein diet, independent of central NPY administration. / Master of Science