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Description
Gene manipulation techniques, such as RNA interference (RNAi), offer a powerful method for elucidating gene function and discovery of novel therapeutic targets in a high-throughput fashion. In addition, RNAi is rapidly being adopted for treatment of neurological disorders, such as Alzheimer's disease (AD), Parkinson's disease, etc. However, a major challenge in both of the aforementioned applications is the efficient delivery of siRNA molecules, plasmids or transcription factors to primary cells such as neurons. A majority of the current non-viral techniques, including chemical transfection, bulk electroporation and sonoporation fail to deliver with adequate efficiencies and the required spatial and temporal control. In this study, a novel optically transparent biochip is presented that can (a) transfect populations of primary and secondary cells in 2D culture (b) readily scale to realize high-throughput transfections using microscale electroporation and (c) transfect targeted cells in culture with spatial and temporal control. In this study, delivery of genetic payloads of different sizes and molecular characteristics, such as GFP plasmids and siRNA molecules, to precisely targeted locations in primary hippocampal and HeLa cell cultures is demonstrated. In addition to spatio-temporally controlled transfection, the biochip also allowed simultaneous assessment of a) electrical activity of neurons, b) specific proteins using fluorescent immunohistochemistry, and c) sub-cellular structures. Functional silencing of GAPDH in HeLa cells using siRNA demonstrated a 52% reduction in the GAPDH levels. In situ assessment of actin filaments post electroporation indicated a sustained disruption in actin filaments in electroporated cells for up to two hours. Assessment of neural spike activity pre- and post-electroporation indicated a varying response to electroporation. The microarray based nature of the biochip enables multiple independent experiments on the same culture, thereby decreasing culture-to-culture variability, increasing experimental throughput and allowing cell-cell interaction studies. Further development of this technology will provide a cost-effective platform for performing high-throughput genetic screens.
ContributorsPatel, Chetan (Author) / Muthuswamy, Jitendran (Thesis advisor) / Helms Tillery, Stephen (Committee member) / Jain, Tilak (Committee member) / Caplan, Michael (Committee member) / Vernon, Brent (Committee member) / Arizona State University (Publisher)
Created2012
Description
The use of electromyography (EMG) signals to characterize muscle fatigue has been widely accepted. Initial work on characterizing muscle fatigue during isometric contractions demonstrated that its frequency decreases while its amplitude increases with the onset of fatigue. More recent work concentrated on developing techniques to characterize dynamic contractions for use in clinical and training applications. Studies demonstrated that as fatigue progresses, the EMG signal undergoes a shift in frequency, and different physiological mechanisms on the possible cause of the shift were considered. Time-frequency processing, using the Wigner distribution or spectrogram, is one of the techniques used to estimate the instantaneous mean frequency and instantaneous median frequency of the EMG signal using a variety of techniques. However, these time-frequency methods suffer either from cross-term interference when processing signals with multiple components or time-frequency resolution due to the use of windowing. This study proposes the use of the matching pursuit decomposition (MPD) with a Gaussian dictionary to process EMG signals produced during both isometric and dynamic contractions. In particular, the MPD obtains unique time-frequency features that represent the EMG signal time-frequency dependence without suffering from cross-terms or loss in time-frequency resolution. As the MPD does not depend on an analysis window like the spectrogram, it is more robust in applying the timefrequency features to identify the spectral time-variation of the EGM signal.
ContributorsAustin, Hiroko (Author) / Papandreou-Suppappola, Antonia (Thesis advisor) / Kovvali, Narayan (Committee member) / Muthuswamy, Jitendran (Committee member) / Arizona State University (Publisher)
Created2012
Description
An understanding of diet habits is crucial in implementing proper management strategies for wildlife. Diet analysis, however, remains a challenge for ruminant species. Microhistological analysis, the method most often employed in herbivore diet studies, is tedious and time consuming. In addition, it requires considerable training and an extensive reference plant collection. The development of DNA barcoding (species identification using a standardized DNA sequence) and the availability of recent DNA sequencing techniques offer new possibilities in diet analysis for ungulates. Using fecal material collected from controlled feeding trials on pygmy goats, (Capra hicus), novel DNA barcoding technology using the P6-loop of the chloroplast trnL (UAA) intron was compared with the traditional microhistological technique. At its current stage of technological development, this study demonstrated that DNA barcoding did not enhance the ability to detect plant species in herbivore diets. A higher mean species composition was reported with microhistological analysis (79%) as compared to DNA barcoding (50%). Microhistological analysis consistently reported a higher species presence by forage class. For affect positive species identification, microhistology estimated an average of 89% correct detection in control diets, while DNA barcoding estimated 50% correct detection of species. It was hypothesized that a number of factors, including variation in chloroplast content in feed species and the effect of rumen bacteria on degradation of DNA, influenced the ability to detect plant species in herbivore diets and concluded that while DNA barcoding opens up new possibilities in the study of plant-herbivore interactions, further studies are needed to standardize techniques and for DNA bar-coding in this context.
ContributorsMurphree, Julie Joan (Author) / Miller, William H. (Thesis advisor) / Steele, Kelly (Committee member) / Salywon, Andrew (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
Description
This study was designed to influence consumer habits, specifically those relating to purchases of fruits, vegetables, and junk food. Previous studies have clearly shown the ineffectiveness of simply describing the health benefits of eating more fruits and vegetables (F/V). In contrast, this study aimed to change the result by changing the message: providing participants with insight into the hidden agendas of food companies and grocery stores, provide useful tips on how to include children when selecting F/V, and emphasizing the importance of parental modeling in regard to food purchases. Participants of this study were separated into two groups, the tour group and the education group. The tour group was guided through a grocery store where they learned about sales tactics and manipulations used by grocery stores and food companies to influence purchases. Education group participants were provided with an education session focusing on USDA and FDA handouts displaying current educational suggestions for increasing F/V consumption. Grocery store receipts were collected and analyzed to track the progress of both groups. The goal of the study was to identify a method of informing consumers that will produce a significant change in behavior. Increasing F/V consumption, even in relatively small amounts, would be an important step forward in improving the diet and overall health of Americans. This study was the first of its kind to measure purchasing patterns objectively (through analysis of purchase receipts, rather than personal opinion/evaluation surveys) and in a wide-scope retail environment that includes all grocery store purchases by participants. Significant increases or decreases in the amount of money spent on F/V, or the amount (pounds) of F/V purchased were not seen, however a small correlation (r = 0.133) exists when comparing the weight of F/V purchased pre/post intervention. Data from Food Frequency Questionnaires shows participants consuming significantly higher amounts of F/V post intervention (p=0.043). The tour group and education group experienced an average increase of 0.7 servings per day. Future interventions might benefit by extending their scope to include cooking demonstrations, in-home interventions, and education on healthy eating outside of the home.
ContributorsKinsfather, Diana (Author) / Johnston, Carol (Thesis advisor) / Hekler, Eric (Committee member) / Tetreault, Colin (Committee member) / Arizona State University (Publisher)
Created2013
Description
Health knowledge alone does not appear to lead to sustained healthy behavior, suggesting the need for alternative methods for improving diet. Recent research shows a possible role of moral contexts of food production on diet related behaviors; however no studies have been conducted to specifically explore the relationship between moral constructs and food consumption. This study examined the relationship between fast food consumption and two measures of morality, Moral Foundations Questionnaire (MFQ), specifically harm/care and purity/sanctity foundations, and the Ethical Concern in food choice (EC) questionnaire, which includes animal welfare, environment protection, political values, and religion subscales. The study also examined the association between the measures of morality. 739 participants, primarily female (71.4%) and non-Hispanic Whites (76.5%), completed an online survey that included the MFQ, the EC questionnaire, and a brief fast food screener. Participant's morality scores in relation to their fast food consumption were examined first using bivariate ANOVA analysis and then using logistic regression to control for covariates. The MFQ foundations were compared with the EC subscales using Pearson correlation coefficient. Significant bivariate relationships were seen between fast food consumption and the MFQ's purity/sanctity foundation and EC's religion subscales (p<0.05). However these significant bivariate relationships did not hold after controlling for gender, race, university education, and religion in the logistic regression analysis. The foundations of the MFQ were positively correlated with the subscales for the EC questionnaire (r values ranging from .233-.613 (p<0.01). MFQ's purity/sanctity foundation and EC's religion subscale were the two most highly correlated (r=.613, p<0.01) showing that moral intuitions may be associated with eating decision making. The study did not find significant associations between MFQ or EC scores and fast food consumption.
ContributorsMartinelli, Sarah (Author) / Ohri-Vachaspati, Punam (Thesis advisor) / Hekler, Eric B. (Committee member) / Wharton, Christopher (Christopher Mack), 1977- (Committee member) / Johnston, Carol (Committee member) / Arizona State University (Publisher)
Created2013
Description
It is widely recognized that dietary protein induces greater satiety compared to carbohydrate and fat. Two separate trials were conducted to assess the use of protein as a dietary approach to manage energy intake (EI). The first, crossover trial, examined 24– hour EI after consuming a high protein bar (HP) vs. a high carbohydrate (HC) bar upon awakening on two separate days and a control, no bar day. Of the 54 participants who entered the trial, 37 subjects completed the study in its entirety. Results showed there was no significant difference in mean EI between the intervention days when the bars were consumed and the control day. The subjects consumed 1752±99 kcal on the control day, and 1846±75 and 1891±110 kcal on the days the HP and HC bars were consumed, respectively (P=0.591). However, compared to the control day, snack bar ingestion was significantly related to an increase in EI for the subjects who self-reported high weekly physical activity levels (n=11) (+22%; P=0.038 and +45%; P=0.030, HP and HC bars, respectively). These data suggest that individuals who have moderate to low physical activity levels compensate for the ingestion of energy bars (regardless of protein content) over a 24–hour period. The second parallel-arm, pilot trial examined the effect of 6 g daily gelatin ingestion vs. control on EI and weight change in healthy, overweight and obese women who initiated a walking program. Of the 37 women who entered the trial, 28 completed the six week trial. The results showed activity level (steps/d) increased in both groups (+ 22%, P=0.022). There was a significant group difference in mean EI at week 6 vs. baseline (–174±612 kcal/d and +197±320 kcal/d, P=0.001; gelatin and control groups, respectively). However, there was no significant between group difference for changes in weight, percent body fat and waist circumference. Those subjects having baseline Disinhibition scores of ≥12 gained significantly more weight throughout the study vs. those scoring <12 (P=0.004). These results indicate that daily gelatin ingestion may be a practical strategy for controlling EI among overweight and obese women initiating an exercise program.
ContributorsTrier, Catherine M (Author) / Johnston, Carol S. (Thesis advisor) / Swan, Pamela D. (Committee member) / Mayol-Kreiser, Sandra N. (Committee member) / Appel, Christy L. (Committee member) / Gaesser, Glenn A. (Committee member) / Arizona State University (Publisher)
Created2012
Description
Sensitivity is a fundamental challenge for in vivo molecular magnetic resonance imaging (MRI). Here, I improve the sensitivity of metal nanoparticle contrast agents by strategically incorporating pure and doped metal oxides in the nanoparticle core, forming a soluble, monodisperse, contrast agent with adjustable T2 or T1 relaxivity (r2 or r1). I first developed a simplified technique to incorporate iron oxides in apoferritin to form "magnetoferritin" for nM-level detection with T2- and T2* weighting. I then explored whether the crystal could be chemically modified to form a particle with high r1. I first adsorbed Mn2+ ions to metal binding sites in the apoferritin pores. The strategic placement of metal ions near sites of water exchange and within the crystal oxide enhance r1, suggesting a mechanism for increasing relaxivity in porous nanoparticle agents. However, the Mn2+ addition was only possible when the particle was simultaneously filled with an iron oxide, resulting in a particle with a high r1 but also a high r2 and making them undetectable with conventional T1-weighting techniques. To solve this problem and decrease the particle r2 for more sensitive detection, I chemically doped the nanoparticles with tungsten to form a disordered W-Fe oxide composite in the apoferritin core. This configuration formed a particle with a r1 of 4,870mM-1s-1 and r2 of 9,076mM-1s-1. These relaxivities allowed the detection of concentrations ranging from 20nM - 400nM in vivo, both passively injected and targeted to the kidney glomerulus. I further developed an MRI acquisition technique to distinguish particles based on r2/r1, and show that three nanoparticles of similar size can be distinguished in vitro and in vivo with MRI. This work forms the basis for a new, highly flexible inorganic approach to design nanoparticle contrast agents for molecular MRI.
ContributorsClavijo Jordan, Maria Veronica (Author) / Bennett, Kevin M (Thesis advisor) / Kodibagkar, Vikram (Committee member) / Sherry, A Dean (Committee member) / Wang, Xiao (Committee member) / Yarger, Jeffery (Committee member) / Arizona State University (Publisher)
Created2012