SUP Program: SOLAR & SURE
1:55-2:15 |
Andorra BastienPresentation Time: 1:55-2:15Home University: Cedar Crest CollegeResearch Mentor: Miriam Braunstein, Microbiology & ImmunologyProgram: SOLAR & SUREResearch Title: Clinically Relevant Bacteriophage Adsorption to Mycobacterium smegmatisMycobacterium abscessus, a non-tuberculosis mycobacteria (NTM), is an emerging threat to immunocompromised patients and those with underlying pulmonary diseases such as cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). M. abscessus infections are extremely challenging to treat due to antibiotic resistance. Bacteriophage (phage) therapy has been shown to target drug-resistant pathogens, posing an alternative to antibiotics. Phages are viruses that infect and kill the bacterial cell host. Much is unknown about the kinetics of phage infection of mycobacteria. Thus, in this study we are determining the timing of phage attachment (adsorption) to the bacterial cell to better understand the period of infection and host cell lysis. The first step in the phage life cycle is phage attachment to bacteria, which is a critical step for treating bacterial infections with phages. Using the model organism Mycobacterium smegmatis and phages BPsΔ33HTH-HRM10, Muddy, and D29 HRM GD40, we demonstrated that the phages display poor adsorption in the time frames that were assayed. Additionally, we found that chloroform kills M. smegmatis but has no effect on the infectivity of the phages tested. In applying chloroform to our adsorption assay, we can more easily distinguish free phages from adsorbed phages. By analyzing phage adsorption, we can assess limitations of specific phages. In the future we can select for phage mutants with increased adsorption rates to optimize phage therapy efficacy. |
Clinically Relevant Bacteriophage Adsorption to Mycobacterium smegmatis | SOLAR & SURE |
2:45-3:05 |
Griffin CarterPresentation Time: 2:45-3:05Home University: University of MiamiResearch Mentor: Craig Cameron, Microbiology and ImmunologyProgram: SOLAR & SUREResearch Title: Nucleic Acid Binding by Enterovirus 2C ProteinAs seen over the past year, RNA viruses are a threat to human health and enteroviruses are no exception. Enterovirus 2C protein is an ATPase that plays an essential role in viral replication and assembly, making it an attractive target for developing antiviral therapeutics. In this work, we use fluorescence polarization to assess potential interactions of 2C protein with fluorescently labeled nucleic acid. We show 2C protein is capable of binding RNA and DNA, although DNA is unable to stimulate 2C ATPase activity. Our initial experiments were performed with poliovirus, but have shown that these observations extend to other enteroviruses, including Coxsackievirus B3, Enterovirus A71, and Enterovirus D68. Understanding the nucleic-acid-binding mechanism is the first step to developing antiviral therapeutics targeting 2C protein with pan-enterovirus activity, thus protecting the world from the threat of enterovirus outbreaks in the future. |
Nucleic Acid Binding by Enterovirus 2C Protein | SOLAR & SURE |
3:10-3:30 |
Camila GarciaPresentation Time: 3:10-3:30Home University: Florida Gulf Coast UniversityResearch Mentor: Brian D. Strahl, Biochemistry & BiophysicsProgram: SOLAR & SUREResearch Title: SANT Domain Mutations that Give Insight to the Histone Binding ActivityThe SAGA(Spt-Ada-Gcn5-Acetyl Transferase) enzymatic complex is a highly conserved complex in eukaryotic organisms from yeast to humans. SAGA is responsible for regulating cellular processes through post-translational modifications(PTMs) on histones (e.g., acetylation, methylation). If malfunctions arise, it can result in the appearance of neurological diseases or cancer. While there’s a great deal of information on the SAGA complex as a whole, the individual contribution of each protein is not well understood, this is specially true for Ada2. Ada2 protein is known for increasing the histone binding activity of the Gnc5 protein, a histone acetylator, however how this happens is unclear. From the Strahl Lab, data showed that the SANT domain of Ada2 binds both H3 and H4. Our goal this summer was to determine which amino acid residues are important for the binding of these histones to the SANT domain. Utilizing point mutations in acidic pockets of the SANT domain, we mutated the amino acids from acidic to basic and to an uncharged amino acid. Once these mutations were in the recombinant plasmid, protein purification, protein pull-down, and western blotting were performed. Preliminary results show that these mutations were not able to eliminate binding of both H3 and H4. Further studies will involve a crystal structure of the current protein to have a view on the exact binding pockets of these histones and mutate those residues. |
SANT Domain Mutations that Give Insight to the Histone Binding Activity | SOLAR & SURE |
3:35-3:55 |
Sy’Keria Destiny GarrisonPresentation Time: 3:35-3:55Home University: Fayetteville State UniversityResearch Mentor: Gregory Scherrer, Department of Cell Biology and PhysiologyProgram: SOLAR & SUREResearch Title: Development of Home Cage Self Administration and Conditioned Place Preference (CPP) Assays in Mice for Addiction StudiesOpioids are the most effective treatments for moderate to severe pain, but they can lead to addiction. The μ-opioid receptor (MOR) mediates various behaviors after morphine exposure including analgesia, hyperalgesia, weight decrease, hyperlocomotion, and addiction. Further investigation of morphine addiction is vital for identifying the specific neuronal populations across the nervous system responsible for addiction. In this study, we generated MOR conditional knockout mice (Chx10Cre/+ ;Oprm1fl/fl) where MOR was removed from the Chx10-expressing neurons, and tested these mice using traditional pain assays after morphine administration. We also established two new behavioral tests, the Conditioned Place Preference (CPP) and the Two Bottle Choice (TBC) assays, to measure addiction in mice. We found that MOR cKO mice showed similar analgesia and morphine withdrawal compared to control littermates, suggesting that the Chx10 expressing neurons may not mediate these effects. Using the CPP assay, mice developed a small preference for morphine after three days of conditioning, however the CPP requires further optimization for addiction testing. We successfully built TBC devices- the experiment is ongoing and we will use it to observe home cage self-administration of opioids in mice. We expect mice to self-administer opioids more than water, with time, during the 14-day experimental period. In the future, we intend to use different MOR cKO mice to elucidate which neurons are involved in the mediation of morphine addiction. |
Development of Home Cage Self Administration and Conditioned Place Preference (CPP) Assays in Mice for Addiction Studies | SOLAR & SURE |
4:00-4:20 |
Jayda YanceyPresentation Time: 4:00-4:20Home University: N.C. A&T State UniversityResearch Mentor: Jeffrey Aube, Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of PharmacyProgram: SOLAR & SUREResearch Title: Triazoles as Kappa Opioid AgonistsThe United States opioid epidemic is caused, in part, due to the available opioid pain relievers having adverse effects like euphoria. One approach to addressing this is to develop an alternative opioid that does not induce side effects. Opioid receptor agonists may activate multiple pathways. The ability of an agonist to only activate one pathway is known as biased agonism. If a biased agonist is developed that only activates the pain-relieving pathway but not the side-effect pathway, one could potentially avoid the undesired side effects of known opioids. Our lab has been working on a class of compounds called triazoles that are biased agonists that function at the kappa opioid receptor through the G-protein pathway. Previous studies have identified triazoles as active, biased compounds, but with low metabolic stability. This work also suggests that sulfur is a site of metabolic instability. Therefore, we hypothesize that a bioisosteric replacement of sulfur with carbon will increase metabolic stability. In this project, we have successfully synthesized an active, carbon-linked triazole that will serve as the basis for future structure activity relationship campaigns. |
Triazoles as Kappa Opioid Agonists | SOLAR & SURE |
4:25-4:45 |
Duyen BuiPresentation Time: 4:25-4:45Home University: UNC-Chapel HillResearch Mentor: Brian Conlon, Microbiology and ImmunologyProgram: SOLAR & SUREResearch Title: Utilizing Palmitoleic Acid to Enhance Bactericidal Activity of Antibiotics in Gram-positive BacteriaLack of commercial viability impedes the development of new antibiotics and the last new class of antibiotic was approved in 2003. Therefore, improving the efficacy of pre-existing antibiotics is a promising strategy to address the antimicrobial resistance crisis. Vancomycin is the frontline antibiotic to treat methicillin-resistant Staphylococcus aureus (MRSA), however its poor bactericidal activity results in high treatment failure. Our research shows that palmitoleic acid, an unsaturated fatty acid, synergizes with vancomycin to kill S. aureus in vitro. However, it is still unknown if palmitoleic acid can also potentiate vancomycin killing of other pathogens. The focus of this research is to test the effectiveness of vancomycin and palmitoleic acid in killing seven other clinically relevant Gram-positive bacteria. We find that palmitoleic acid enhances the bactericidal activity of vancomycin against several Gram-positive species. These findings support the use of unsaturated fatty acids as an antibiotic adjuvant for the treatment of bacterial infections and provides evidence for its use as a novel strategy to enhance our current arsenal of antibiotics. Moving forward, the aim of the project will unravel the underlying mechanism of synergy to identify other antibiotic adjuvants. |
Utilizing Palmitoleic Acid to Enhance Bactericidal Activity of Antibiotics in Gram-positive Bacteria | SOLAR & SURE |