Theses and Dissertations

Reducing the amount of error and introduced data variability increases the accuracy of Western blot results. In this study, different methods of normalization for loading differences and data alignment were explored with respect to their impact on Western blot results. GAPDH was compared to the LI-COR Revert total protein stain as a loading control. The impact of normalizing data to a control condition, which is commonly done to align Western blot data distributed over several immunoblots, was also investigated. Specifically, this study addressed whether normalization to a small subset of distinct controls on each immunoblot increases pooled data variability compared to a larger set of controls. Protein expression data for NOX-2 and SOD-2 from a study investigating the protective role of the bradykinin type 1 receptor in angiotensin-II induced left ventricle remodeling were used to address these questions but are also discussed in the context of the original study. The comparison of GAPDH and Revert total protein stain as a loading control was done by assessing their correlation and comparing how they affected protein expression results. Additionally, the impact of treatment on GAPDH was investigated. To assess how normalization to different combinations of controls influences data variability, protein data were normalized to the average of 5 controls, the average of 2 controls, or an average vehicle and the results by treatment were compared. The results of this study demonstrated that GAPDH expression is not affected by angiotensin-II or bradykinin type 1 receptor antagonist R-954 and is a less sensitive loading control compared to Revert total protein stain. Normalization to the average of 5 controls tended to reduce pooled data variability compared to 2 controls. Lastly, the results of this study provided preliminary evidence that R-954 does not alter the expression of NOX-2 or SOD-2 to an expression profile that would be expected to explain the protection it confers against Ang-II induced left ventricle remodeling.

This thesis identifies and explains two main problems students face during their internships. The first problem relates to feeling bored at internships due to the simplicity of projects or lack of work. From the interviews conducted, several strategies to avoid this boredom were created, including having employers design education plans for interns to further their knowledge in programs such as excel during their downtime. The second problem with internships discovered focuses on the gap between what is taught in schools versus what is expected of interns in practice. This thesis identifies several opportunities for improvement in education and strategies on how to handle feeling overwhelmed on intern projects due to lack of knowledge.

Over 40% of adults in the United States are considered obese. Obesity is known to cause abnormal metabolic effects and lead to other negative health consequences. Interestingly, differences in metabolism and contractile performance between obese and healthy weight individuals are associated with differences in skeletal muscle fiber type composition between these groups. Each fiber type is characterized by unique metabolic and contractile properties, which are largely determined by the myosin heavy chain isoform (MHC) or isoform combination that the fiber expresses. In previous studies, SDS-PAGE single fiber analysis has been utilized as a method to determine MHC isoform distribution and single fiber type distribution in skeletal muscle. Herein, a methodological approach to analyze MHC isoform and fiber type distribution in skeletal muscle was fine-tuned for use in human and rodent studies. In the future, this revised methodology will be implemented to evaluate the effects of obesity and exercise on the phenotypic fiber type composition of skeletal muscle.

Since 1975, the prevalence of obesity has nearly tripled around the world. In 2016, 39% of adults, or 1.9 billion people, were considered overweight, and 13% of adults, or 650 million people, were considered obese. Furthermore, Cardiovascular disease remains to be the leading cause of death for adults in the United States, with 655,000 people dying from related conditions and consequences each year. Including fiber in one’s dietary regimen has been shown to greatly improve health outcomes in regards to these two areas of health. However, not much literature is available on the effects of corn-based fiber, especially detailing the individual components of the grain itself. The purpose of this preliminary study was to test the differences in influence on both LDL-cholesterol and triglycerides between treatments based on whole-grain corn flour, refined corn flour, and 50% refined corn flour + 50% corn bran derived from whole grain cornmeal (excellent fiber) in healthy overweight (BMI ≥ 25.0 kg/m2) adults (ages 18 - 70) with high LDL cholesterol (LDL ≥ 120mg/dL). 20 participants, ages 18 - 64 (10 males, 10 females) were involved. Data was derived from blood draws taken before and after each of the three treatments as well as before and after each treatment’s wash out periods. A general linear model was used to assess the effect of corn products on circulating concentrations of LDL-cholesterol and triglycerides. From the model, it was found that the whole-grain corn flour and the 50% refined corn flour + 50% corn bran drive from whole grain cornmeal treatments produced a higher, similar benefit in reductions in LDL-cholesterol. However, the whole grain flour, refined flour, and bran-based fiber treatments did not influence the triglyceride levels of the participants throughout this study. Further research is needed to elucidate the effects of these fiber items on cardiometabolic disease markers in the long-term as well as with a larger sample size.

Motor learning is the process of improving task execution according to some measure of performance. This can be divided into skill learning, a model-free process, and adaptation, a model-based process. Prior studies have indicated that adaptation results from two complementary learning systems with parallel organization. This report attempted to answer the question of whether a similar interaction leads to savings, a model-free process that is described as faster relearning when experiencing something familiar. This was tested in a two-week reaching task conducted on a robotic arm capable of perturbing movements. The task was designed so that the two sessions differed in their history of errors. By measuring the change in the learning rate, the savings was determined at various points. The results showed that the history of errors successfully modulated savings. Thus, this supports the notion that the two complementary systems interact to develop savings. Additionally, this report was part of a larger study that will explore the organizational structure of the complementary systems as well as the neural basis of this motor learning.

Mutations in the DNA of somatic cells, resulting from inaccuracies in DNA<br/>replication or exposure to harsh conditions (ionizing radiation, carcinogens), may be<br/>loss-of-function mutations, and the compounding of these mutations can lead to cancer.<br/>Such mutations can come in the form of thymine dimers, N-𝛽 glycosyl bond hydrolysis,<br/>oxidation by hydrogen peroxide or other radicals, and deamination of cytosine to uracil.<br/>However, many cells possess the machinery to counteract the deleterious effects of<br/>such mutations. While eukaryotic DNA repair enzymes decrease the incidence of<br/>mutations from 1 mistake per 10^7 nucleotides to 1 mistake per 10^9 nucleotides, these<br/>mutations, however sparse, are problematic. Of particular interest is a mutation in which<br/>uracil is incorporated into DNA, either by spontaneous deamination of cysteine or<br/>misincorporation. Such mutations occur about one in every 107 cytidine residues in 24<br/>hours. DNA uracil glycosylase (UDG) recognizes these mutations and cleaves the<br/>glycosidic bond, creating an abasic site. However, the rate of this form of DNA repair<br/>varies, depending on the nucleotides that surround the uracil. Most enzyme-DNA<br/>interactions depend on the sequence of DNA (which may change the duplex twist),<br/>even if they only bind to the sugar-phosphate backbone. In the mechanism of uracil<br/>excision, UDG flips the uracil out of the DNA double helix, and this step may be<br/>impaired by base pairs that neighbor the uracil. The deformability of certain regions of<br/>DNA may facilitate this step in the mechanism, causing these regions to be less<br/>mutable. In DNA, base stacking, a form of van der Waals forces between the aromatic<br/>nucleic bases, may make these uracil inclusions more difficult to excise. These regions,<br/>stabilized by base stacking interactions, may be less susceptible to repair by<br/>glycosylases such as UDG, and thus, more prone to mutation.