MSU Student Research Celebration
Permanent URI for this collectionhttps://scholarworks.montana.edu/handle/1/405
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Item TRIMmunity and MAGE Interactions(Montana State Univeristy, 2017-04) Crane, BrittneeInterferon (IFN)-β is involved in immune responses against viral infections. Some TRIM proteins, such as TRIM5 and TRIM22, are IFN-stimulated genes (ISGs) since their expression levels increases in response to IFNβ-intitiated signaling and, interestingly, have been shown to have direct and indirect antiviral activities, respectively. In a recent transciptomic analysis of polarized macrophages, we found that TRIM31 was specifically upregulated in response to INFβ treatment relative to the 32 additional activation conditions tested. We hypothesize that TRIM31 has antiviral activity, a role that may be dependent on formation of active E3 ubiquitin ligase complexes containing melanoma antigen gene (MAGE) proteins. We have initiated yeast-two hybrid-based experiments to confirm interactions between TRIM31 and three members of the MAGE family as well as to identify TRIM31 interaction partners. Co-localization studies will follow. The overall aim of our research program will be to characterize TRIM31 and other TRIM proteins to shed light on this family of proteins that has been subjected to strong, positive evolutionary pressure.Item Analyzing the Effects of Treating Human Lung Cell Tissue with a Low-Level Chronic Metal Mixture(Montana State Univeristy, 2017-04) Paolini, MorganIn 1983, Butte, Montana was designated a Superfund site due to the impact of over a century of mining activity. A recent study was conducted analyzing the accumulation of 36 elements in the hair and 11 elements in the blood of Butte residents as opposed to a control population, as well as an analysis of the soil and air of the Butte area. Elevated levels of several metals, including Cu, Mn, and As, were found in the hair, blood, soil, and air. The focus of this study was on the elevated metals in the air, arsenic and manganese. Previous studies have shown that elevated levels of metals, such as Pb, As, and Mn, can evoke serious health effects such as neurodegenerative diseases and cancers. In order to investigate the toxicity of airborne metal ions on Butte residents, this study replicated the environmental exposure of the airborne metals found in the human study, and evaluated the biological response of normal lung cells. To do this, human lung cell tissue (BEAS-2B) was treated with a low-level chronic metal mixture of either NaAsO2, MnCl2·4H2O, or a mixture of the two metals. Following metal exposure, the cell cultures were assessed with XTT cell viability assays measuring cell viability based on mitochondrial respiration and gene array plates measuring genetic changes. The results of these tests are expected to provide preliminary knowledge regarding the health impacts of low-level chronic metal exposure effects on human lung cells.Item Growth of ZnTe Nanowires for Observation with Femtosecond Pump-Probe Microscopy(Montana State Univeristy, 2017-04) Smyser, KoriIn an effort to contribute to the search for alternative fuel sources, we propose to examine the excited state behavior of piezoelectric ZnTe nanowires, which have applications as mechanical energy harvesters. Our initial work has focused on designing a replicable synthesis for chemical vapor deposition (CVD) growth of uniform ZnTe nanowires. With success, we now begin an ultrafast spectroscopic study of the carrier dynamics in ZnTe nanowires.Item Metabalomics Profiles of Ciproflaxacin Treated Staphylococcus aureus and Acinetobacter baumannii Biofilms(Montana State University, 2017-04) Scott, LiamMany bacterial organisms have the ability to form complex conglomerations of cells known as biofilms. It is important to understand this stage of bacterial life because it facilitates colonization and persistence in environments not suitable for planktonic cells. For example, biofilm structures grant an inherent resistance to antibiotic agents, a trait that is particularly concerning for infectious bacteria such as Staphylococcus aureus and Acinetobacter baumannii. With bacterial organisms such as these displaying powerful resistance to antibiotics, it is becoming ever more critical to understand biofilm structures, along with the inherent antibiotic resistance that they provide. To begin to understand these structures, many researchers have turned to analysis of the metabolomic profiles of bacterial species, and how those profiles differ between planktonic cells and biofilm conglomerations. In this project, it is proposed that the metabolomic profiles of S. aureus and A. baumannii will differ significantly between not only their free floating and biofilm states, but also in their antibiotic treated biofilm states. As such, I will work to combine an external metabolite analysis conducted using gas chromatography mass spectrometry (GCMS), with an internal metabolite analysis conducted using liquid chromatography mass spectrometry (LCMS), to create metabolomic profiles of each variant of these bacteria listed above. With this data, it will be possible to elucidate the biochemical pathways critical to S. aureus and A. baumannii biofilm antibiotic resistance. On a larger scale, this information will be critical in the pursuit of sensitizing S. aureus and A. baumannii to modern antibiotics.Item Regulating T-cell Responses: Screening and Characterization of Possible Kinase Inhibitors in Ex vivo/In vitro Models(Montana State University, 2017-04) Mitchell, Pete T.Protein phosphorylation is a key component of T-Cell Receptor (TCR) signaling pathways. Upon activation of a TCR, protein kinases, such as Lck, phosphorylate proteins downstream of the cascade, which results in recruitment of scaffold and adaptor proteins. Transcription factors are modulated following this cascade, which may lead to proliferation and production of cytokines as part of an immune response to the activation of TCR mediated signal transduction. However, when the transcriptional regulation goes awry in lymphocytes, immune dysfunction can occur. Over- or auto-reactive lymphocytes resulting from intracellular pathway disruptions can lead to disorders, such as chronic inflammation, diabetes, and cancer. One approach to manage immune dysfunction is by targeting the deregulated activation of protein kinases and their cascades. The objective of this project is to screen selected sesquiterpene lactones (SLs) for anti-proliferative activity and low cytotoxic effects, followed by characterizing of any affected protein kinases in murine and human models. Cells were pre-treated with a compound for 20 minutes, activated with anti-CD3/CD28, and then T-cell proliferative assays or ELISA were employed. From our experimentation, Erk1/2 was shown to have dose-dependent phosphorylation and was used to gauge other SLs for Erk1/2 selectivity. Additionally, the mechanism of the Erk1/2-SL interaction (estafiatine) was investigated. The data are still preliminary and more experiments are needed to draw conclusions, in particular to the mechanism.Item A Divergent Synthesis of Spongistatin(Montana State University, 2017-04) McGlamery, DevinSince the dawn of organic chemistry, natural products have long been coveted for their remarkable complexity as well as unmatched bio-activities. Spongistatin is one such natural product possessing activity against human melanoma with a GI50 value of just 25 pM. In addition to medicinal potential, spongistatin is an intriguing synthetic puzzle, containing a multitude of intricate stereocenters as well as two spiroketal subunits that join the A,B and C,D ring systems forming the spitoketal moiety’s of the molecule. Spongistatin’s two spiroketals exist as two stereoisomers, the axial-axial (A,B) and the axial-equatorial (C,D). The axial-axial spiroketal is favored due to stabilization via the double anomeric effect, whereupon the ring system is stabilized by lengthening of the C-O bond by n→σ * donation. However due to poor orbital overlap the axial-equatorial stereoisomer does not benefit from double stabilization. The Cook group has developed a divergent synthesis of highly substituted spiroketals via an ortholactone allylsilane fragment coupling reaction which proceeds through a Sakauri type mechanism. To make this more applicable to spongistatin we aim to develop methodology around less rigid frameworks (scheme 1). We have so far focused on the synthesis of cyclic ortholactones from the C4 unsubstituted dihydropyran, via an oxidative catalytic nucleopalladation reaction. Regretfully the cyclic nature of the molecule provides an increase in stability that prevents the operation of the previously developed spiroketalization conditions (scheme 1). To combat this, recent work has focused on generation of methoxy or ethoxy substituted ortholactones, in order to generate the C4 unsubstituted spiroketals (scheme 2).Item Assessing evolutionary pressure on Herpes Simplex Virus Type 1 genomes during infection and spread(Montana State University, 2017-04) Law, GabrielleThis project will quantify evolutionary pressure, through mutation and selection, of herpes simplex virus type 1 (HSV-1) during replication and spread. HSV-1 infection involves replication and spread in neurons, in part contributing to HSV-1 as the leading cause of viral encephalitis in developed countries. HSV-1 is a dsDNA virus with an accordingly low point mutation rate. Yet a high level of viral genome diversity has been observed across populations. Viral diversity is produced through the combined effects of evolutionary pressure and selection on HSV-1 genomes. We are establishing methods to quantify viral population diversity and the effects of evolutionary pressure on HSV-1 genomes. We quantified inter-viral genomic recombination using a marker transfer assay to understand the impact of selection pressures, such as neuron replication, on viral diversity. The marker transfer assay utilized two unique, comparably fit viral isolates with genetically distant fluorescent protein expressing cassettes to co-infect cells. We scored viral progeny as either parent or recombinant based on marker expression as a percentile of the population. We adapted the marker transfer assay for both in vitro and in vivo HSV-1 infections. We utilized the HSV murine eye model to quantify the effect of spread and replication in neurons on viral populations. Understanding how HSV responds to antiviral selections during replication and spread will enhance current treatment for herpes infection and provide insight for vaccine developments.Item Real Time Metabolomics and Applied Dynamics(Montana State University, 2017-04) LaCroix, IanMetabolite Extraction Chip Mass Spectrometry (MEC-MS) is an emerging technique that allows near real-time monitoring of complex fluids. Here, we show that MEC-MS can be used to monitor the metabolism of an E. coli culture grown in a chemostat bioreactor. Periodic samples of live E. coli cells were automatically pulsed through the MEC-MS, over the course of twenty-four hours. The significance of the data produced by MEC-MS was that; it provided a continuous description of how the metabolic profile of E. coli changed during the sample period. This type of data has previously lacked in metabolomics studies. We used the data to develop a continuous model of differential equations, with the goal to explain the dynamics of metabolism in E. coli. The next step will include; using MEC-MS to monitor E. coli under oxidative stress conditions, and then to analyze the metabolic dynamics. With an aim to detect biomarkers of oxidative stress in a predictive fashion. Future work will include; application of this methodology to the analysis of human biofluids in order to detect novel biomarkers of disease.Item Identification of Biomarkers for Tumor Development in the Liver using Bovine Serum Albumin as a Molecular Sensor(Montana State Univeristy, 2017-04) Budeski, KatherineAnalyzing biological fluids using untargeted metabolomics has proved challenging because it is time-consuming and, at times, inconclusive. As an alternative method, the Bothner Lab has established the use of the protein bovine serum albumin (BSA) as a molecular sensor to differentiate complex biological solutions. Serum albumin is the most abundant protein in mammalian blood, where it transports lipids, hormones and drugs. The natural role of BSA as a carrier protein inspired the Bothner Lab to test this proteins’ ability to bind a wide variety of small molecules with a goal of targeted metabolite extractions from complex biological samples. This method has been coined the protein sensor assay (PSA). A new set of methods has been developed to perform metabolite extractions using the PSA on serum collected from mice with induced liver cancer from the Schmidt Lab at Montana State University. The Schmidt Lab has done much work to identify the key steps in biochemical pathways like oxidative stress in mice with induced cancer. In this project I have used the PSA together with liquid chromatography mass spectrometry and statistical analyses to identify biomarkers for tumor development in mouse serum. In the future, the PSA may be used to speed up analysis and improve confidence in disease diagnostics.Item Further Investigation of How and Why the Lipid Bilayer Composition of Escherichia coli (E. coli) Differs in Aerobic and Anaerobic Environments(Montana State Univeristy, 2017-04) Paris, NinaEscherichia coli (E. coli) is a highly studied bacterium because it is easy to grow and can grow in aerobic and anaerobic conditions. E. coli cells are surrounded by a lipid bilayer to help facilitate what can enter and leave the cell and to help protect it from the environment. Lipids are very sensitive to energy levels in a cell and therefore can give insight into metabolic stress in cells. Last summer I determined that the lipid composition of E. coli lipid membranes is different when grown in aerobic versus anaerobic conditions. Experiments since then have been done and it has been determined that the lipid composition changes quickly once E. coli cells are transitioned from aerobic to anaerobic environments. This shows that lipid synthesis and metabolism of the lipid bilayer of E. coli is affected by whether oxygen is available.