Volume 3, No. 1, 2017
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Acta Spartae is the undergraduate journal of sciences and mathematics at The University of Tampa, and is designed to recognize and promote the vibrant undergraduate research community at UT by providing a forum for the dissemination of research and ideas produced at the University.
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Item Editor's Note(College of Natural and Health Sciences, The University of Tampa, 2017) Ethan Deneault; CNHSItem Stability of p53 mRNA Isoforms in MCF7 Cells(College of Natural and Health Sciences, The University of Tampa, 2017) Zachary M. Connelly; L. Michael CarastroTumor protein 53 (p53) is a tumor suppressor gene that has two key functions. This protein regulates cell cycle and induces apoptosis, or programmed cell death. TP53 mRNA isoforms differ in lengths of the 5’-leader sequence. Longer isoforms (p53 mRNAL) contain a putative upstream open reading frame, not present in shorter 5’ leaders (p53 mRNA-S). We hypothesize p53 mRNAL is subject to nonsense-mediated mRNA decay (NMD). Treatment with cycloheximide, caffeine, and wortmannin diminish NMD. Our objective was to chemically inhibit NMD in MCF7 cells concurrently treated with Actinomycin D. Cellular proteins were subjected to SDSPAGE and western analyses for p53. Isolated RNA samples were synthesized into cDNA, then subjected to qRT-PCR analyses of p53 mRNA isoforms. p53 mRNA-L/ p53 mRNA-S isoform ratios (L/S) were calculated from Relative Quantification (RQ) values obtained from p53 mRNA isoforms, by comparing treated to untreated samples and were reported as mean L/S ratios and standard deviations. Actinomycin D treatment, without inhibitors, resulted in a L/S = 1.070 ( 0.05). Actinomycin D co-treatment with cycloheximide, caffeine or wortmannin resulted in L/S means of 1.159 ( 0.07), 1.181 ( 0.18) and 1.279 ( 0.15), respectively. Western blot analyses were consistent with reduced translation of p53 protein in cycloheximide treated cells. Caffeine and wortmannin treated cells contained a prominent p53 protein band consistent with hypo-phosphorylated p53. In conclusion, chemical treatment effectively inhibited translation and kinase activity. p53 mRNA-L is partially rescued in cells treated with inhibitors of translation and kinase activity.Item Occurrence of MRSA on Fish in the Hillsborough River(College of Natural and Health Sciences, The University of Tampa, 2017) Zachery Rivera, Kimberly Grassett; Ann Williams; Bridgette FroeschkeMRSA is a drug-resistant strain of Staphylococcus aureus known to cause frequent infections in humans. Water quality assessments have identified MRSA at varying levels throughout the Hillsborough River which indicates that MRSA may be present on the organisms that reside in those waters. In this experiment, Fish were collected from multiple sites throughout the Hillsborough River and swabbed to assess the presence of MRSA on the organism. Bacterial samples were grown on Mannitol Salt Agar, isolated, and tested for drugresistance using the Kirby Bauer Disk Diffusion Assay. MRSA was isolated from the scales and inside the mouths of fish at multiple sites on the Hillsborough River. The presence of MRSA on these fish could potentially pose a health risk to fisherman who encounter the fish harboring MRSA isolates. Fisherman with an open wound on their hand that handle these fish could acquire a MRSA infection and require subsequent medical treatment as a result.Item Quantum mechanical investigation of the inner sphere reduction of the [(NSSSN)Co(III)Cl+2] cation and its analogs(College of Natural and Health Sciences, The University of Tampa, 2017) William Smith; Thomas Jackman; Olaseni SodeThe inner sphere pathway is an electron transfer (ET) mechanism that utilizes a bridging ligand to covalently link oxidant and reductant centers. The reductions of chloro-N-methyl-bis(5-amino- 3-thiapentyl)amine cobalt(III) [(NSNSN)Co(III)Cl+3] and chloro(1,11- diamino-3,6,9-trithiaundecane)cobalt(III) cation [(NSSSN)Co(III)Cl+2] by iron(II) via inner sphere ET have been shown experimentally to occur with rate constants more than 107 times faster than the nitrogen analog [(NNNNN)Co(III)Cl+2]. It has been hypothesized that this is due to non-bridging ligand effects. To test this hypothesis, the role of ground state electronic effects by the sulfur-containing ligands on the ET is investigated through the use of quantum chemistry methods. The non-bridging ligand effects were explored through the structural parameters of the cobalt complexes and by examining the LUMOs using both wavefunction theory (WFT) and density functional theory (DFT) methods. We show that the complexes containing sulfur atoms (NSSSN and NSNSN) display similar geometries. These are in contrast to the nitrogen analog (NNNNN) geometry, pointing towards a possible structural driving force in the rate constant difference.Item Transformation of a Mixed Probiotic Culture and Escherichia coli B with the Antibiotic Resistant Plasmid, pGLO(College of Natural and Health Sciences, The University of Tampa, 2017) Raven M. Wright, Haley L. Thompson; Eric FreundtProbiotics are microorganisms residing in the gastrointestinal tract that have been shown to help with metabolic functions, immune responses, and in the prevention of pathogenic diseases. In this study, we sought to determine if the probiotic brand, Garden of Life, could acquire novel phenotypes through heat-shock transformation. We predicted that Garden of Life, along with Escherichia coli HB101 and Escherichia coli B would be able to grow on ampicillin enriched plates after transformation with the plasmid, pGLO, which contained the gene for beta-lactamase. During the study, all three bacterial cultures underwent heat shock transformation in the presence of pGLO. After transformation was complete, each new culture was plated on LB agar containing ampicillin and arabinose to determine whether antibiotic resistance and GFP expression were observed. The results showed that only Escherichia coli HB101 had successfully become antibiotic resistant. Our results demonstrate that the gram positive species in Garden of Life probiotics are not able to efficiently take up plasmid DNA using heat shock transformation and that the method appeared to have strain restrictions as well.Item Microbial Properties of Montipora Coral Mucus(College of Natural and Health Sciences, The University of Tampa, 2017) Jayde A. Zimmerman, Cody J. Cox; Eric FreundtMucus produced by corals has a varied understanding with many gaps. Although there are many hypotheses regarding the function and purpose of the mucus itself, and the many microbial communities that inhabit it, there are still many inquiries about it. The purpose of this experiment was to determine whether coral mucus possessed diverse microbes and whether the mucus held any antibacterial properties against gram positive or gram negative bacteria compared to seawater. Mucus was collected from Montipora species and plated on marine agar. Individual bacterial colonies were isolated and gram stained. Our results showed that the mucus does not show antibacterial activity but contains as much as five times the amount of bacteria in comparison to seawater. Additionally, the coral mucus contained exclusively gram negative bacteria whereas the seawater contained both gram negative and gram positive bacteria. These results suggest that coral mucus is a hospitable site for growth of gram negative bacteria.