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Bacteria have been shown to possess a large array of regulatory mechanisms to not just respond to a diverse array of environmental stresses, but to injurious artificial proteins as well. A previous investigation introduced DX, a man-made ATP sequestering protein into Escherichia coli (E. coli) which resulted in the formation of novel endoliposome structures and induced a viable but non-culturable state (VBNC) that was not easily reversed. It was hypothesized that the broadly conserved bacterial stringent response pathway may have been responsible for the observed phenotypic changes. With the goal of unveiling the molecular mechanism behind this novel response, changes in cellular morphology and physiology upon DX expression were assessed in a population of E. coli encoding a dysfunctional relA gene, one of the two genes controlling the induction of the stringent response. It was ultimately shown that RelA directly contributed to cellular filamentation, endoliposome structure formation, and the induction of a VBNC state. While the stringent response has been extensively shown to induce a VBNC state, to our knowledge, relA has not yet been shown to induce filamentation or coordinate the formation of endoliposome structures in bacteria. As the stringent response has been shown to be increasingly involved in antibiotic tolerance, this study provided an exciting opportunity to further characterize this adaptive response pathway to aid in the future development of novel therapeutics. In addition to this, this study continued to highlight that the DX protein may serve one of the first tools to allow for the direct selection of bacteria in a VBNC state by morphologically distinguishing non-culturable cells through cellular filamentation.
ContributorsFrost, Fredrick Charles (Author) / Chaput, John (Thesis director) / Wachter, Rebekka (Committee member) / Korch, Shaleen (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-12
Description
The Roller Derby Club at Arizona State University became a student organization in the fall of 2013. They became a practicing team known as the Derby Devils in the spring of 2014. This project documents the creation and development a collegiate roller derby team as they go from a student organization to an athletic team. Collegiate roller derby is still in its infant stages and therefore the purpose of this project is to provide a guide for future collegiate roller derby teams as well as other athletic teams.
ContributorsLee, Alisa Yulim (Author) / Looser, Devoney (Thesis director) / Hultsman, Wendy (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / Department of Chemistry and Biochemistry (Contributor) / W. P. Carey School of Business (Contributor)
Created2014-12
Description
Repeating tiles made of DNA were used to try to form an indefinitely large structure. Both the tiles and structure were 2D. Two different patterns were tested, one corrugated and one not. Corrugation means that the tiles alternated between facing up and facing down, canceling out any curvature to the tile and creating a slightly corrugated but largely 2D pattern. Annealing methods were also experimented with. Annealing the structure in two, separate steps as opposed to one was tested. Another experiment was comparing cyclic versus linear annealing. A linear decrease in temperatures defines the linear annealing, and a cyclic method involved a linear drop to a certain temperature, followed by a slight increase in temperature and cooling back down again. This cycle is done several times before it continues linear cool down. It was seen that both corrugated and non-corrugated structures could be made. In both cases tiles that make up a larger section of the overall pattern were more successful. This is especially important for the non-corrugated pattern. Linear and 2step annealing methods seem to yield the best results.
ContributorsHunt, Ashley Elizabeth (Author) / Yan, Liu (Thesis director) / Yan, Hao (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
Continuing work has been done on a novel class of anti-cancer drugs employing a vinylogous extended amidine system functionalized into benzimidazole ring. Three new derivatives, utilizing a true sugar mimic at the N1-position, have been synthesized. Compounds 6-amino-1-[2-(2-hydroxyethoxy)ethyl-4-imino-2,5-dimethyl-1H-benzimidazole- 7-one (5), 6-amino-1-[2-(2-hydroxysulfonoethoxy)ethyl-4-imino-2,5-dimethyl-1H-benzimidazole-7-one (6), and 6-amino-1-[2-(2-hydroxysulfonoethoxy)ethyl-4-methylimino-2,5-dimethyl-1H-benzimidazole-7-one (7) have been synthesized utilizing similar protocols used in the synthesis of previously screened anti-tumor drugs produced by this laboratory. Compounds (5) and (6) have undergone screening similar to the National Cancer Institute’s (NCIs) Developmental Therapeutic Program (DTP), performed by Dr. Dan LaBarbera at the University of Boston. Both compounds show high cytotoxicity, with complete cell death at 5 µM and bioactive concentrations in the low nanomolar concentrations; more complete data is forthcoming. The proposed mechanism of action is through inhibition of p90RSK 1-2 which is responsible for the phosphorylation of Bcl-2 associated death promoter (BAD), a key metabolite in directing the onset of apoptosis. Future directions of next generation derivatives include modifying the 2-position of the benzimidazole ring into a halogenating or alkylating agent and possibly replacing the methanesulfonate with a phosphate group. This research is being published in the Journal of Medicinal Chemistry.
ContributorsMorrison, Zachary Tyler (Author) / Skibo, Edward (Thesis director) / Lefler, Scott (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
Description
Photosynthesis is a critical process that fixes the carbon utilized in cellular respiration. In higher plants, the immutans gene codes for a protein that is both involved in carotenoid biosynthesis and plastoquinol oxidation (Carol et al 1999, Josse et al 2003). This plastoquinol terminal oxidase (PTOX) is of great interest in understanding electron flow in the plastoquinol pool. In order to characterize this PTOX, polyclonal antibodies were developed. Expression of Synechococcus WH8102 PTOX in E. coli provided a useful means to harvest the protein required for antibody production. Once developed, the antibody was tested for limit of concentration, effectiveness in whole cell lysate, and overall specificity. The antibody raised against PTOX was able to detect as low as 10 pg of PTOX in SDS-PAGE, and could detect PTOX extracted from lysed Synechococcus WH8102. The production of this antibody could determine the localization of the PTOX in Synechococcus.
ContributorsKhan, Mohammad Iqbal (Author) / Moore, Thomas (Thesis director) / Redding, Kevin (Committee member) / Roberson, Robert (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Life Sciences (Contributor)
Created2014-05
Description
Overexpression of AVP1 (Arabidopsis vacuolar pyrophosphatase), a type I H+ pyrophosphatase, results in greater biomass, possibly due to a function in sucrose transport within the phloem. Overexpression of the phloem lipid-associated family protein (PLAFP) was shown to increase the number of vascular bundles in Arabidopsis. Could these two phenotypes complement one another additively? In this work, double mutants overexpressing both AVP1 and PLAFP were characterized. These double mutants have enhanced biomass, greater leaf area, and a larger number of vascular bundles than the single mutant lines. Overexpression of PLAFP does not result in any increase in rhizosphere acidification capacity.
ContributorsWilson, Sean (Co-author) / Furstenau, Tara (Co-author) / Gaxiola, Roberto (Thesis director) / Mason, Hugh (Committee member) / Wojciechowski, Martin (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Life Sciences (Contributor)
Created2014-05
Description
Translating research has been a goal of the Department of Health and Human Services since 1999. Through two years of iteration and interview with our community members, we have collected insights into the barriers to accomplishing this goal. Liberating Science is a think-tank of researchers and scientists who seek to create a more transparent process to accelerate innovation starting with behavioral health research.
ContributorsRaghani, Pooja Sioux (Author) / Hekler, Eric (Thesis director) / Buman, Matthew (Committee member) / Pruthi, Virgilia Kaur (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Biomedical Informatics Program (Contributor)
Created2014-05
Description
There are many different methods of learning the guitar and teaching it to oneself. I explored classical methods as well as learned by tablature. I compared it to how I have learned the piano and other things in past such as languages and sports. This paper narrates the process but also give advise for someone teaching themself any subject matter. Here are the main points: (1) Choose something you are passionate about (2) Use what you already know, and apply strategies that have worked before (3) Try learning the subject in different ways (4) Build a strong foundation of fundamentals (5) Take advantage of various resources (6) Do not be afraid to make mistakes (7) Perseverance will pay off. I found that I preferred the tablature method but learned a lot from both. I plan on continuing to improve at both methods and want to expand my learning to songwriting.
ContributorsCoelet, Collin Patrick (Author) / Schildkret, David (Thesis director) / Halick, Mary (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
Description
Workplace injuries, especially those caused by overexertion, are a significant issue among firefighters. This investigation filmed 8 firefighters during simulated firefighting tasks. A detailed movement analysis for each task was completed and used to infer the prime muscles and injury-prone joints during each task. A comprehensive strength training program was developed from this information to help reduce workplace injuries in firefighters.
ContributorsAbel, Morgan Daniel (Author) / Hinrichs, Richard (Thesis director) / Harper, Erin (Committee member) / Barrett, The Honors College (Contributor) / School of Nutrition and Health Promotion (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
Description
The focus of this project was to look at alternative treatments for endocrine resistant breast cancer (ERBC), which are breast cancers that have become resistant to hormone therapies such as Tamoxifen or aromatase inhibitors. The first part of this project involves investigating the relationship between histone de-acetylase inhibitor Vorinostat and Tamoxifen in MCF7 G11 cells, Tamoxifen resistant sub-clones, according to the PSOC Time grant. The second part involves targeting the androgen receptor (AR) in MCF7 sub-clones with AR antagonists, Bicalutamide and MDV3100, and investigating the possible usage of AR as a biomarker, due to over-expression of AR in ERBC, in accordance with the Mayo ASU Seed Grant.
The synergistic effects between Vorinostat and Tamoxifen observed through a phase II study on breast cancer patients resistant to hormone therapy may involve more than the modulation of ER-alpha to reverse Tamoxifen resistance in ERBC cells. RT-qPCR of genes expressed in Tamoxifen resistant cells, trefoil factor 1(TFF1) and v-myc avian myelocytomatosis viral oncogene homolog (MYC), were evaluated along with ESR1 and Diablo as a control. MYC was observed to have increased expression in the treated cells, whereas the other genes had a decrease in their expression levels after the cells were treated for 3 days with Vorinostat IC30 of 1 µM. As for targeting the AR, MCF7 Tamoxifen sensitive and resistant cells were not affected by the AR antagonists to determine an IC50. The cell viability for all MCF7 sub-clones only decreased for high concentrations of 5.56 µM - 50 µM in Bicalutamide and 16.67 µM – 50 µM of MDV1300. Furthermore, hormone depletion of MCF7 G11 Tamoxifen resistant sub-clones did not show a great response to DHT stimulation or the AR antagonists. In the RT-qPCR, the MCF7 G11 cells showed an increase in mRNA expression for ER, AR, and PR after 4 hours of treatment with estradiol. As for the DHT treatment, ER, AR, PR, and PSA had a minimal increase in the fold change, but the fold change in AR was less than in the estradiol treatment. The Mayo Clinic will investigate the possible usage of AR as a biomarker through immunohistochemistry.
The synergistic effects between Vorinostat and Tamoxifen observed through a phase II study on breast cancer patients resistant to hormone therapy may involve more than the modulation of ER-alpha to reverse Tamoxifen resistance in ERBC cells. RT-qPCR of genes expressed in Tamoxifen resistant cells, trefoil factor 1(TFF1) and v-myc avian myelocytomatosis viral oncogene homolog (MYC), were evaluated along with ESR1 and Diablo as a control. MYC was observed to have increased expression in the treated cells, whereas the other genes had a decrease in their expression levels after the cells were treated for 3 days with Vorinostat IC30 of 1 µM. As for targeting the AR, MCF7 Tamoxifen sensitive and resistant cells were not affected by the AR antagonists to determine an IC50. The cell viability for all MCF7 sub-clones only decreased for high concentrations of 5.56 µM - 50 µM in Bicalutamide and 16.67 µM – 50 µM of MDV1300. Furthermore, hormone depletion of MCF7 G11 Tamoxifen resistant sub-clones did not show a great response to DHT stimulation or the AR antagonists. In the RT-qPCR, the MCF7 G11 cells showed an increase in mRNA expression for ER, AR, and PR after 4 hours of treatment with estradiol. As for the DHT treatment, ER, AR, PR, and PSA had a minimal increase in the fold change, but the fold change in AR was less than in the estradiol treatment. The Mayo Clinic will investigate the possible usage of AR as a biomarker through immunohistochemistry.
ContributorsVorachitti, Merica (Author) / LaBaer, Joshua (Thesis director) / Anderson, Karen (Committee member) / Gonzalez, Laura (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05