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Background: Measles virus (MV) infections are the main cause of vaccine-preventable death in children younger than 5 years. The World Health Organization (WHO) has estimated there are over 20 million cases of measles every year. Currently, diagnostic methods rely on enzyme immunoassays (EIA) to detect IgM or IgG Abs in serum. These commercial assays measure reactivity against the immunodominant N antigen and can have a false negative rates of 20-30%. Centralized testing by clinical labs can delay rapid screening in an outbreak setting. This study aims to develop a rapid molecular diagnostic assay to detect IgG reactive to five individual MV proteins representing 85% of the measles proteome. Methods: MV genes were subcloned into pANT_cGST vector to generate C-terminal GST fusion proteins. Single MV cistrons were expressed using in vitro transcription/translation (IVTT) with human cell lysate. Expression of GST-tagged proteins was measured using a sandwich ELISA for GST expression using relative light units (RLUs) as readouts. Single MV antigens were used as bait to determine the IgG-dependent reactivity in 12 serum samples obtained from immunized animals with previously determined neutralization titer (NT) and the correlation between NT and ELISA reactivity was determined. Results: Protein expression of five measles genes of interest, M, N, F, H, and L, was measured. L exhibited the strongest protein expression with an average RLU value of 4.34 x 10^9. All proteins were expressed at least 50% greater than control (2.33 x 10^7 RLU). As expected, reactivity against the N was the highest, followed by reactivity against M, F, H and L. The best correlation with NT titer was reactivity against F (R^2 = 0.62). Conclusion: These data indicate that the expression of single MV genes M, N, F, H, and L are suitable antigens for serologic capture analysis of measles immunity.
ContributorsMushtaq, Zuena (Author) / Anderson, Karen (Thesis director) / Reyes del Valle, Jorge (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
Rubisco is a very important protein which catalyzes the addition of CO2 to ribulose-1,5-bisphosphate (RuBP) to form two molecules of 3-phosphoglycerate in photosynthesis. Rubisco activase is the protein which functions to uninhibit Rubisco, however proof of a physical interaction has never been shown. A possible method for determining the interaction of the two proteins is by Förster Resonance Energy Transfer (FRET) based analysis of the two proteins. Attempts to get a FRET signal from these two proteins have been unsuccessful. To get better results, Ficoll 70, a crowding agent, was used. Analysis suggests that Ficoll 70 does not affect the fluorescence of Alexa-fluor 488 and Alexa-fluor 647 used to label the two proteins. Further analysis also suggests that while the Alexa label on Rubisco activase does not affect the ATPase activity of the protein, the protein also does not have a high rate of ATP turnover.
ContributorsTichacek, Laura Renee (Author) / Wachter, Rebekka (Thesis director) / Levitus, Marcia (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
Hydrogen has the potential to be a highly efficient fuel source. Its current production via steam reformation of natural gas, however, consumes a large amount of energy and gives off carbon dioxide. A newer method has since surfaced: using a microorganism's metabolism to drive hydrogen production. In this study, the conditions for maximum hydrogen production in Heliobacterium modesticaldum were identified and assessed. The cells were grown under varying conditions and their headspaces were sampled using a gas chromatogram to measure the amount of accumulated hydrogen during each condition. Two cell batches were grown under nitrogen-fixing conditions (-NH4+), while the other two cell batches were grown under non-nitrogen-fixing conditions (+NH4+). The headspaces were then exchanged with either nitrogen (N2) or argon (Ar2). It was found that the condition for which the most hydrogen was produced was when the cells were grown under nitrogen-fixing conditions and the headspace was exchanged with argon. These results suggest that most of Heliobacteria modesticaldum's hydrogen production is due to nitrogenase activity rather than hydrogenase activity. Further research is recommended to quantify the roles of nitrogenase, [NiFe] hydrogenase, and [FeFe] hydrogenase.
ContributorsMcmahon, Savanah Dior (Author) / Redding, Kevin (Thesis director) / Ghirlanda, Giovanna (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Music (Contributor)
Created2015-05
Description
With higher than normal levels of cardiovascular disease, diabetes, obesity, and sexually transmitted diseases within the Deaf community, it is clear that the current level of medical care is failing. This can stem from many different issues, from the mistrust of doctors to the fact that scientific education in the Deaf community is introduced at a later time in comparison to the hearing population. Similarly, there is a distinct lack of Deaf and hard of hearing medical staff. All of this culminates in the fact that the system needs to change. The addition of more Deaf staff in the medical environment, more staff interpreters, and doctors that have experience with the Deaf community engenders the metamorphosis of this group from unhealthy to healthy. Extra exposure for doctors during clinical rotations to deaf patients as well as training in Deaf Culture will further increase the comfort level of Deaf community members utilizing health services. As a result, the overall state of medical care for the Deaf population will improve with these modifications to the current system.
ContributorsBrar, Serbnoor Singh (Author) / Quinn, Paul (Thesis director) / Howard, Pamela (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
This paper focuses on the Szemerédi regularity lemma, a result in the field of extremal graph theory. The lemma says that every graph can be partitioned into bounded equal parts such that most edges of the graph span these partitions, and these edges are distributed in a fairly uniform way. Definitions and notation will be established, leading to explorations of three proofs of the regularity lemma. These are a version of the original proof, a Pythagoras proof utilizing elemental geometry, and a proof utilizing concepts of spectral graph theory. This paper is intended to supplement the proofs with background information about the concepts utilized. Furthermore, it is the hope that this paper will serve as another resource for students and others to begin study of the regularity lemma.
ContributorsByrne, Michael John (Author) / Czygrinow, Andrzej (Thesis director) / Kierstead, Hal (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
F2-isoprostanes are a series of prostaglandin-like compounds derived from the free radical-mediated lipid peroxidation of arachidonic acid, a polyunsaturated fatty acid that is ubiquitously expressed in cell membranes. F2-isoprostanes are biomarkers of oxidative stress, an imbalance between oxidants and antioxidants that can cause damage to DNA, proteins, lipids, and carbohydrates. Increased production of lipid peroxidation products have been implicated in the pathology of a number of conditions and diseases in humans. The objective of this thesis was to (1) optimize the LC/MS/MS F2-isoprostane method currently used in human samples for use in research animals and veterinary medicine, including the use of solid phase extraction, and (2) validate the optimized method in rodent and canine experimental studies. Our optimized method showed that Lyprinol treatment in dogs with osteoarthritis decreases F2-isoprostane levels nearly 2-fold. In addition, adjuvant alpha-tocopherol prevented tumor-induced increased F2-isoprostane levels. Finally, contrary to earlier studies using less specific ELISA F2-isoprostane methods, we demonstrate that unconditioned dogs benefit from low intensity exercise. Our data demonstrate successful optimization of the human LC/MS/MS F2-isoprostane method in rats and canines. Importantly, our results emphasize the need to use the more sensitive and specific LC/MS/MS method as compared to ELISA-based assays in order to distinguish the 15- and 5-series F2-isoprostanes, evidenced in particular by the two canine studies.
ContributorsCorrigan, Devin Connell (Author) / Redding, Kevin (Thesis director) / Anderson, Karen (Committee member) / Mustacich, Debbie (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
This study assessed the effects of running an eating prevention program on body image satisfaction/behavior and the leadership skills of collegiate women. The sample included a group of 43 undergraduate women who voluntarily chose to become peer-educators in the eating prevention program called the Body Project. Self-report questionnaires evaluating both the preoccupation with personal body image and general leadership skills were distributed and collected electronically. The results were analyzed to determine that being a peer leader in the Body project did not increase eating disorder symptoms but actually decreased the symptoms. It was also determined that being a peer educator had no effect on leadership skills. Therefore, being a peer leader is beneficial for reducing eating disorder symptoms, but not for advancing leadership skills.
ContributorsCamiliere, Taylor Marie (Author) / Perez, Marisol (Thesis director) / Cavanaugh Toft, Carolyn (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Department of Psychology (Contributor)
Created2015-05
Description
Smartphone-based healthcare. It's becoming more real as technology advances, adding value to healthcare and opening the doors to more personalized medicine. The Medical Pal is a smartphone application that can track symptoms and analyze trends in the severity of those symptoms, alerting healthcare providers when there is a significant increase in the symptom severity. This is especially directed to the palliative patient, whose care is focused on managing symptoms and providing comfort. The HIPAA-compliant server used for the smartphone application was Catalyze.io and 40 Mayo Clinic Arizona palliative patients were surveyed on their smartphone usage to test the acceptance of this app in a clinical setting. A trial involving 9 simulated patients was conducted over a two week period to test the functionality of the app. A majority of surveyed patients (85%) expressed favor for the idea of a mobile ESAS, and the app was functioning, with the capability of displaying patient data on a healthcare provider's account. This project is intentionally a door-opener to an open field of opportunity for mobile health, symptom observation, and improvements in healthcare delivery.
ContributorsDao, Lelan Diep (Author) / Cortese, Denis (Thesis director) / Lipinski, Christopher (Committee member) / Fitch, Tom (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
The field of art conservation has evolved parallel to scientific advancements and in response to changing social conditions. Methods have expanded from dusting the surface of a work to bombarding an image with x-rays in an effort to fulfill the ultimate goal of restoration: to return a piece of art to the state originally created by the artist. The process has become much easier and more complex with the introduction of analytical equipment that shifted the focus of the field away from maintenance towards academic study. This work illustrates the changes the field and demonstrates the application of scientific techniques to specific works.
ContributorsSanders, Hailey Jane (Author) / Fahlman, Besty (Thesis director) / Codell, Julie (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
The 23-step total synthesis of dolastatin 16, a cyclic depsipeptide of marine origin, is presented. Included are syntheses of nonnatural amino acids dolamethylleuine and dolaphenvaline. The biological activity of the synthetic product differed from naturally isolated dolastatin 16, which may indicate the initial screening identified an inactive compound and the active one was not detected initially, or may be a result of the conformational dynamics induced by the proline residues. Additionally, a family of structural analogues to the bacillistatins, another cyclic marine depsipeptide, were synthesized. These were deemed the silstatins. 8 modifications were produced. The alterations aimed to introduce a heteroatomic residue for further derivatization, such as producing an antibody-drug conjugate. This introduction did in general decrease the neoplastic activity of these agents, as expected, but by modulating the lipophilicity of the compound we were able to salvage much of the potency of the bacillistatins while potentially allowing prodrug development.
ContributorsMacdonald, Christian B. (Author) / Pettit, George (Thesis director) / Wang, Xu (Committee member) / Melody, Noeleen (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Mathematical and Statistical Sciences (Contributor)
Created2015-05