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- Genre: Academic theses
- Creators: Johnston, Carol
Description
Synechocystis sp PCC 6803 is a photosynthetic cyanobacterium that can be easily transformed to produce molecules of interest; this has increased Synechocystis’ popularity as a clean energy platform. Synechocystis has been shown to produce and excrete molecules such as fatty acids, isoprene, etc. after appropriate genetic modification. Challenges faced for large–scale growth of modified Synechocystis include abiotic stress, microbial contamination and high processing costs of product and cell material. Research reported in this dissertation contributes to solutions to these challenges. First, abiotic stress was addressed by overexpression of the heat shock protein ClpB1. In contrast to the wild type, the ClpB1 overexpression mutant (Slr1641+) tolerated rapid temperature changes, but no difference was found between the strains when temperature shifts were slower. Combination of ClpB1 overexpression with DnaK2 overexpression (Slr1641+/Sll0170+) further increased thermotolerance. Next, we used a Synechocystis strain that carries an introduced isoprene synthase gene (IspS+) and that therefore produces isoprene. We attempted to increase isoprene yields by overexpression of key enzymes in the methyl erythritol phosphate (MEP) pathway that leads to synthesis of the isoprene precursor. Isoprene production was not increased greatly by MEP pathway induction, likely because of limitations in the affinity of the isoprene synthase for the substrate. Finally, two extraction principles, two–phase liquid extraction (e.g., with an organic and aqueous phase) and solid–liquid extraction (e.g., with a resin) were tested. Two–phase liquid extraction is suitable for separating isoprene but not fatty acids from the culture medium. Fatty acid removal required acidification or surfactant addition, which affected biocompatibility. Therefore, improvements of both the organism and product–harvesting methods can contribute to enhancing the potential of cyanobacteria as solar–powered biocatalysts for the production of petroleum substitutes.
ContributorsGonzalez Esquer, Cesar Raul (Author) / Vermaas, Willem (Thesis advisor) / Chandler, Douglas (Committee member) / Bingham, Scott (Committee member) / Nielsen, David (Committee member) / Arizona State University (Publisher)
Created2013
Description
The body is capable of regulating hunger in several ways. Some of these hunger regulation methods are innate, such as genetics, and some, such as the responses to stress and to the smell of food, are innate but can be affected by body conditions such as BMI and physical activity. Further, some hunger regulation methods stem from learned behaviors originating from cultural pressures or parenting styles. These latter regulation methods for hunger can be grouped into the categories: emotion, environment, and physical. The factors that regulate hunger can also influence the incidence of disordered eating, such as eating in the absence of hunger (EAH). Eating in the absence of hunger can occur in one of two scenarios, continuous EAH or beginning EAH. College students are at a particularly high risk for EAH and weight gain due to stress, social pressures, and the constant availability of energy dense and nutrient poor food options. The purpose of this study is to validate a modified EAH-C survey in college students and to discover which of the three latent factors (emotion, environment, physical) best predicts continual and beginning EAH. To do so, a modified EAH-C survey, with additional demographic components, was administered to students at a major southwest university. This survey contained two questions, one each for continuing and beginning EAH, regarding 14 factors related to emotional, physical, or environmental reasons that may trigger EAH. The results from this study revealed that the continual and beginning EAH surveys displayed good internal consistency reliability. We found that for beginning and continuing EAH, although emotion is the strongest predictor of EAH, all three latent factors are significant predictors of EAH. In addition, we found that environmental factors had the greatest influence on an individual's likelihood to continue to eat in the absence of hunger. Due to statistical abnormalities and differing numbers of factors in each category, we were unable to determine which of the three factors exerted the greatest influence on an individual's likelihood to begin eating in the absence of hunger. These results can be utilized to develop educational tools aimed at reducing EAH in college students, and ultimately reducing the likelihood for unhealthy weight gain and health complications related to obesity.
ContributorsGoett, Taylor (Author) / Johnston, Carol (Thesis advisor) / Lee, Chong (Committee member) / Lespron, Christy (Committee member) / Arizona State University (Publisher)
Created2013
Description
In October, 2009, participants of the Arizona Special Supplemental Nutrition Program for Women, Infants and Children (WIC) began receiving monthly Cash Value Vouchers (CVV) worth between six and 10 dollars towards the purchase of fresh fruits and vegetables. Data from the Arizona Department of Health Services (ADHS) showed CVV redemption rates in the first two years of the program were lower than the national average of 77% redemption. In response, the ADHS WIC Food List was expanded to also include canned and frozen fruits and vegetables. More recent data from ADHS suggest that redemption rates are improving, but variably exist among different WIC sub-populations. The purpose of this project was to identify themes related to the ease or difficulty of WIC CVV use amongst different categories of low-redeeming WIC participants. A total of 8 focus groups were conducted, four at a clinic in each of two Valley cities: Surprise and Mesa. Each of the four focus groups comprised one of four targeted WIC participant categories: pregnant, postpartum, breastfeeding, and children with participation ranging from 3-9 participants per group. Using the general inductive approach, recordings of the focus groups were transcribed, hand-coded and uploaded into qualitative analysis software resulting in four emergent themes including: interactions and shopping strategies, maximizing WIC value, redemption issues, and effect of rule change. Researchers identified twelve different subthemes related to the emergent theme of interactions and strategies to improve their experience, including economic considerations during redemption. Barriers related to interactions existed that made their purchase difficult, most notably anger from the cashier and other shoppers. However, participants made use of a number of strategies to facilitate WIC purchases or extract more value out of WIC benefits, such as pooling their CVV. Finally, it appears that the fruit and vegetable rule change was well received by those who were aware of the change. These data suggest a number of important avenues for future research, including verifying these themes are important within a larger, representative sample of Arizona WIC participants, and exploring strategies to minimize barriers identified by participants, such as use of electronic benefits transfer-style cards (EBT).
ContributorsBertmann, Farryl M. W (Author) / Wharton, Christopher (Christopher Mack), 1977- (Thesis advisor) / Ohri-Vachaspati, Punam (Committee member) / Johnston, Carol (Committee member) / Hampl, Jeffrey (Committee member) / Dixit-Joshi, Sujata (Committee member) / Barroso, Cristina (Committee member) / Arizona State University (Publisher)
Created2013
Description
There are several visual dimensions of food that can affect food intake, example portion size, color, and variety. This dissertation elucidates the effect of number of pieces of food on preference and amount of food consumed in humans and motivation for food in animals. Chapter 2 Experiment 1 showed that rats preferred and also ran faster for multiple pieces (30, 10 mg pellets) than an equicaloric, single piece of food (300 mg) showing that multiple pieces of food are more rewarding than a single piece. Chapter 2 Experiment 2 showed that rats preferred a 30-pellet food portion clustered together rather than scattered. Preference and motivation for clustered food pieces may be interpreted based on the optimal foraging theory that animals prefer foods that can maximize energy gain and minimize the risk of predation. Chapter 3 Experiment 1 showed that college students preferred and ate less of a multiple-piece than a single-piece portion and also ate less in a test meal following the multiple-piece than single-piece portion. Chapter 3 Experiment 2 replicated the results in Experiment 1 and used a bagel instead of chicken. Chapter 4 showed that college students given a five-piece chicken portion scattered on a plate ate less in a meal and in a subsequent test meal than those given the same portion clustered together. This is consistent with the hypothesis that multiple pieces of food may appear like more food because they take up a larger surface area than a single-piece portion. All together, these studies show that number and surface area occupied by food pieces are important visual cues determining food choice in animals and both food choice and intake in humans.
ContributorsBajaj, Devina (Author) / Phillips, Elizabeth D. (Thesis advisor) / Cohen, Adam (Committee member) / Johnston, Carol (Committee member) / Bimonte-Nelson, Heather A. (Committee member) / Arizona State University (Publisher)
Created2013
Description
Dietary protein is known to increase postprandial thermogenesis more so than carbohydrates or fats, probably related to the fact that amino acids have no immediate form of storage in the body and can become toxic if not readily incorporated into body tissues or excreted. It is also well documented that subjects report greater satiety on high- versus low-protein diets and that subject compliance tends to be greater on high-protein diets, thus contributing to their popularity. What is not as well known is how a high-protein diet affects resting metabolic rate over time, and what is even less well known is if resting metabolic rate changes significantly when a person consuming an omnivorous diet suddenly adopts a vegetarian one. This pilot study sought to determine whether subjects adopting a vegetarian diet would report decreased satiety or demonstrate a decreased metabolic rate due to a change in protein intake and possible increase in carbohydrates. Further, this study sought to validate a new device called the SenseWear Armband (SWA) to determine if it might be sensitive enough to detect subtle changes in metabolic rate related to diet. Subjects were tested twice on all variables, at baseline and post-test. Independent and related samples tests revealed no significant differences between or within groups for any variable at any time point in the study. The SWA had a strong positive correlation to the Oxycon Mobile metabolic cart but due to a lack of change in metabolic rate, its sensitivity was undetermined. These data do not support the theory that adopting a vegetarian diet results in a long-term change in metabolic rate.
ContributorsMoore, Amy (Author) / Johnston, Carol (Thesis advisor) / Appel, Christy (Thesis advisor) / Gaesser, Glenn (Committee member) / Arizona State University (Publisher)
Created2012
Description
In somatic cells, the mitotic spindle apparatus is centrosomal and several isoforms of Protein Kinase C (PKC) have been associated with the mitotic spindle, but their role in stabilizing the mitotic spindle is unclear. Other protein kinases such as, Glycogen Synthase Kinase 3â (GSK3â) also have been shown to be associated with the mitotic spindle. In the study in chapter 2, we show the enrichment of active (phosphorylated) PKCæ at the centrosomal region of the spindle apparatus in metaphase stage of 3T3 cells. In order to understand whether the two kinases, PKC and GSK3â are associated with the mitotic spindle, first, the co-localization and close molecular proximity of PKC isoforms with GSK3â was studied in metaphase cells. Second, the involvement of inactive GSK3â in maintaining an intact mitotic spindle was shown. Third, this study showed that addition of a phospho-PKCæ specific inhibitor to cells can disrupt the mitotic spindle microtubules. The mitotic spindle at metaphase in mouse fibroblasts appears to be maintained by PKCæ acting through GSK3â. The MAPK pathway has been implicated in various functions related to cell cycle regulation. MAPKK (MEK) is part of this pathway and the extracellular regulated kinase (ERK) is its known downstream target. GSK3â and PKCæ also have been implicated in cell cycle regulation. In the study in chapter 3, we tested the effects of inhibiting MEK on the activities of ERK, GSK3â, PKCæ, and á-tubulin. Results from this study indicate that inhibition of MEK did not inhibit GSK3â and PKCæ enrichment at the centrosomes. However, the mitotic spindle showed a reduction in the pixel intensity of microtubules and also a reduction in the number of cells in each of the M-phase stages. A peptide activation inhibitor of ERK was also used. Our results indicated a decrease in mitotic spindle microtubules and an absence of cells in most of the M-phase stages. GSK3â and PKCæ enrichment were however not inhibited at the centrosomes. Taken together, the kinases GSK3â and PKCæ may not function as a part of the MAPK pathway to regulate the mitotic spindle.
ContributorsChakravadhanula, Madhavi (Author) / Capco, David G. (Thesis advisor) / Chandler, Douglas (Committee member) / Clark-Curtiss, Josephine (Committee member) / Newfeld, Stuart (Committee member) / Arizona State University (Publisher)
Created2012
Description
The F1Fo ATP synthase is required for energy conversion in almost all living organisms. The F1 complex is a molecular motor that uses ATP hydrolysis to drive rotation of the γ–subunit. It has not been previously possible to resolve the speed and position of the γ–subunit of the F1–ATPase as it rotates during a power stroke. The single molecule experiments presented here measured light scattered from 45X91 nm gold nanorods attached to the γ–subunit that provide an unprecedented 5 μs resolution of rotational position as a function of time. The product of velocity and drag, which were both measured directly, resulted in an average torque of 63±8 pN nm for the Escherichia coli F1-ATPase that was determined to be independent of the load. The rotational velocity had an initial (I) acceleration phase 15° from the end of the catalytic dwell, a slow (S) acceleration phase during ATP binding/ADP release (15°–60°), and a fast (F) acceleration phase (60°–90°) containing an interim deceleration (ID) phase (75°–82°). High ADP concentrations decreased the velocity of the S phase proportional to 'ADP-release' dwells, and the F phase proportional to the free energy derived from the [ADP][Pi]/[ATP] chemical equilibrium. The decreased affinity for ITP increased ITP-binding dwells by 10%, but decreased velocity by 40% during the S phase. This is the first direct evidence that nucleotide binding contributes to F1–ATPase torque. Mutations that affect specific phases of rotation were identified, some in regions of F1 previously considered not to contribute to rotation. Mutations βD372V and γK9I increased the F phase velocity, and γK9I increased the depth of the ID phase. The conversion between S and F phases was specifically affected by γQ269L. While βT273D, βD305E, and αR283Q decreased the velocity of all phases, decreases in velocity due to βD302T, γR268L and γT82A were confined to the I and S phases. The correlations between the structural locations of these mutations and the phases of rotation they affect provide new insight into the molecular basis for F1–ATPase γ-subunit rotation.
ContributorsMartin, James (Author) / Frasch, Wayne D (Thesis advisor) / Chandler, Douglas (Committee member) / Gaxiola, Roberto (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2012
Description
Objective: Vinegar consumption studies have demonstrated possible therapeutic effects in reducing HbA1c and postprandial glycemia. The purpose of the study was to closely examine the effects of a commercial vinegar drink on daily fluctuations in fasting glucose concentrations and postprandial glycemia, and on HbA1c, in individuals at risk for Type 2 Diabetes Mellitus (T2D). Design: Thirteen women and one man (21-62 y; mean, 46.0±3.9 y) participated in this 12-week parallel-arm trial. Participants were recruited from a campus community and were healthy and not diabetic by self-report. Participants were not prescribed oral hypoglycemic medications or insulin; other medications were allowed if use was stable for > 3 months. Subjects were randomized to one of two groups: VIN (8 ounces vinegar drink providing 1.5 g acetic acid) or CON (1 vinegar pill providing 0.04 g acetic acid). Treatments were taken twice daily immediately prior to the lunch and dinner meals. Venous blood samples were drawn at trial weeks 0 and 12 to measure insulin, fasting glucose, and HbA1c. Subjects recorded fasting glucose and 2-h postprandial glycemia concentrations daily using a glucometer. Results: The VIN group showed significant reductions in fasting capillary blood glucose concentrations (p=0.05) that were immediate and sustained throughout the duration of the study. The VIN group had reductions in 2-h postprandial glucose (mean change of −7.6±6.8 mg/dL over the 12-week trial), but this value was not significantly different than that for the CON group (mean change of 3.3±5.3 mg/dL over the 12-week trial, p=0.232). HbA1c did not significantly change (p=0.702), but the reduction in HbA1c in the VIN group, −0.14±0.1%, may have physiological relevance. Conclusions: Significant reductions in HbA1c were not observed after daily consumption of a vinegar drink containing 1.5 g acetic acid in non-diabetic individuals. However, the vinegar drink did significantly reduce fasting capillary blood glucose concentrations in these individuals as compared to a vinegar pill containing 0.04 g acetic acid. These results support a therapeutic effect for vinegar in T2D prevention and progression, specifically in high-risk populations.
ContributorsQuagliano, Samantha (Author) / Johnston, Carol (Thesis advisor) / Appel, Christy (Committee member) / Dixon, Kathleen (Committee member) / Arizona State University (Publisher)
Created2013
Description
Objective: The purpose of this randomized parallel arm trial was to demonstrate the effects of daily fish oil supplementation (600mg per day for eight weeks) on body composition and body mass in young healthy women, aged 18-38, at a large southwestern university. Design: 26 non-obese (mean BMI 23.7±0.6 kg/m2), healthy women (18-38y; mean, 23.5±1.1 y) from a southwestern Arizona university campus community completed the study. Subjects were healthy, non-smokers, consuming less than 3.5 oz of fish per week according to self-report. Participants were randomized to one of two groups: FISH (600 mg omega-3 fatty acids provided in one gel capsule per day), or CON (1000 mg coconut oil placebo provided in one gel capsule per day). Body weight, BMI, and percent body fat were measured using a stadiometer and bioelectrical impedance scale at the screening visit and intervention weeks 1, 4, and 8. 24-hour dietary recalls were also performed at weeks 1 and 8. Results: 8 weeks of omega-3 fatty acid supplementation did not significantly alter body weight (p=0.830), BMI (p=1.00), or body fat percentage (p=0.600) as compared to placebo. Although not statistically significant, 24-hour dietary recalls performed at the beginning and end of the intervention revealed a trend towards increased caloric intake in the FISH group and decreased caloric intake in the CON group throughout the course of the study (p=0.069). If maintained, this difference in caloric intake could have physiological relevance. Conclusions: Omega-3 fatty acids do not significantly alter body weight or body composition in healthy young females. These findings do not refute the current recommendations for Americans to consume at least 8 oz of omega-3-rich seafood per week, supplying 250 mg EPA and DHA per day. More research is needed to investigate the potential for omega-3 fatty acids to modulate daily caloric intake.
ContributorsTeran, Bianca (Author) / Johnston, Carol (Thesis advisor) / Johnson, Melinda (Committee member) / Ohri-Vachaspati, Punam (Committee member) / Arizona State University (Publisher)
Created2013
Description
In 2002, a scientifically derived food guide pyramid for vegetarians, the Modified Food Guide for Lacto-ovo-vegetarians and Vegans was published and well received. Now that 10 years have passed, new scientific literature regarding the bioavailability of the nutrients of key concern in vegetarian diets has been published, and the graphical format of the nation's food guide has evolved from a pyramid shape into a circular plate. The objective of this research was to examine the post-2002 literature regarding the bioavailability of key nutrients in vegetarian diets; to use this information to update the recommendations made in the 2002 Modified Food Guide Pyramid for Lacto-ovo-vegetarians and Vegans; and to adapt this revised food plan to the new USDA MyPlate format. This process involved reviewing the scientific literature to determine if the DRIs for the nutrients of key concern in vegetarian diets are adequate for the vegetarian population and using this information to develop new recommendations for vegetarians if necessary, analyzing the nutrient content of representative foods in different food groups, reconfiguring the food groups so that foods with like nutrient components were grouped together, determining the number of servings of each food group required to meet vegetarians' nutrient requirements at three caloric levels, and developing sample menus. A circular plate graphic, the Vegetarian Plate, was designed to illustrate the recommendations of this updated food guide. This updated, scientifically derived food guide provides a sound base for diet planning for lacto-ovo-vegetarians and vegans. Further research is needed to assess the Vegetarian Plate's adequacy for children, pregnant and lactating women, athletes, and individuals with medical conditions or chronic diseases.
ContributorsFladell, Lauren (Author) / Johnston, Carol (Thesis advisor) / Vaughan, Linda (Committee member) / Shepard, Christina (Committee member) / Arizona State University (Publisher)
Created2013